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Sample records for 2-dimensional polyacrylamide gel

  1. Electroblotting from Polyacrylamide Gels.

    PubMed

    Goldman, Aaron; Ursitti, Jeanine A; Mozdzanowski, Jacek; Speicher, David W

    2015-11-02

    Transferring proteins from polyacrylamide gels onto retentive membranes is now primarily used for immunoblotting. A second application that was quite common up to about a decade ago was electroblotting of proteins for N-terminal and internal sequencing using Edman chemistry. This unit contains procedures for electroblotting proteins from polyacrylamide gels onto a variety of membranes, including polyvinylidene difluoride (PVDF) and nitrocellulose. In addition to the commonly used tank or wet transfer system, protocols are provided for electroblotting using semidry and dry systems. This unit also describes procedures for eluting proteins from membranes using detergents or acidic extraction with organic solvents for specialized applications.

  2. Drying SDS-Polyacrylamide Gels.

    PubMed

    Sambrook, Joseph; Russell, David W

    2006-09-01

    INTRODUCTIONThis protocol describes a method for drying SDS-polyacrylamide gels. Gels containing proteins radiolabeled with (35)S-labeled amino acids must be dried before autoradiographic images can be obtained. Nonradioactive gels can also be preserved by drying.

  3. SDS-Polyacrylamide Gel Electrophoresis of Proteins.

    PubMed

    Sambrook, Joseph; Russell, David W

    2006-09-01

    INTRODUCTIONThis protocol describes the separation of proteins by SDS-polyacrylamide gel electrophoresis. SDS is used with a reducing agent and heat to dissociate the proteins. SDS-polypeptide complexes form and migrate through the gels according to the size of the polypeptide. By using markers of known molecular weight, the molecular weight of the polypeptide chain(s) can be estimated.

  4. Enhanced detection of glycoproteins in polyacrylamide gels.

    PubMed

    Muñoz, G; Marshall, S; Cabrera, M; Horvat, A

    1988-05-01

    A highly sensitive and simple method to enhance detection of glycoproteins resolved by either one- or two-dimensional polyacrylamide gel electrophoresis is described. The method is a modification of the procedure described by D. Fargeaud et al. (D. Fargeaud, J. C. Benoit, F. Kato, and G. Chappuis (1984) Arch. Virol. 80, 69-82) that uses concanavalin A conjugated with fluorescein isothyocyanate to detect the carbohydrate moiety of glycoproteins. Briefly, the electrophoresed gel is exposed to the fluorescent lectin, thoroughly washed, and sequentially transferred to 50% methanol in deionized water and to absolute methanol. The result is an abrupt dehydration of the gel which turns evenly white and stiff. At least a twofold enhancement of fluorescence is obtained as detected by exposing the treated gel to an appropriate uv source. The sensitivity of the procedure allows us to detect purified immunoglobulin molecules by their carbohydrate content in the range of 0.2 microgram of total protein. The specificity of the detection is demonstrated by a comparison with the corresponding polypeptide profile obtained by silver nitrate staining of the gel.

  5. A method for horizontal polyacrylamide slab gel electrophoresis.

    PubMed

    Bellomy, G R; Record, M T

    1989-01-01

    We present a simplified method of preparation of polyacrylamide gels which is totally analogous to the procedure now widely used to pour and run horizontal agarose gels. The acrylamide is poured into an open air gel mold consisting of a glass plate with a masking tape border and a comb. It is subsequently run in a submarine horizontal electrophoresis apparatus. The electrophoretic mobility and resolution of DNA fragments obtained in such gels are identical to results obtained with gels poured and run in the vertical configuration. Numerous advantages of horizontal polyacrylamide gel electrophoresis are discussed.

  6. Electrophoresis of DNA in agarose gels, polyacrylamide gels and in free solution.

    PubMed

    Stellwagen, Nancy C

    2009-06-01

    This review describes the electrophoresis of curved and normal DNA molecules in agarose gels, polyacrylamide gels and in free solution. These studies were undertaken to clarify why curved DNA molecules migrate anomalously slowly in polyacrylamide gels but not in agarose gels. Two milestone papers are cited, in which Ferguson plots were used to estimate the effective pore size of agarose and polyacrylamide gels. Subsequent studies on the effect of the electric field on agarose and polyacrylamide gel matrices, DNA interactions with the two gel matrices, and the effect of curvature on the free solution mobility of DNA are also described. The combined results suggest that the anomalously slow mobilities observed for curved DNA molecules in polyacrylamide gels are primarily due to preferential interactions of curved DNAs with the polyacrylamide gel matrix; the restrictive pore size of the matrix is of lesser importance. In free solution, DNA mobilities increase with increasing molecular mass until leveling off at a plateau value of (3.17 +/- 0.01) x 10(-4) cm2/V s in 40 mM Tris-acetate-EDTA buffer at 20 degrees C. Curved DNA molecules migrate anomalously slowly in free solution as well as in polyacrylamide gels, explaining why the Ferguson plots of curved and normal DNAs containing the same number of base pairs extrapolate to different mobilities at zero gel concentration.

  7. UNIT 10.7 Electroblotting from Polyacrylamide Gels

    PubMed Central

    Goldman, Aaron; Speicher, David W.

    2015-01-01

    Transferring proteins from polyacrylamide gels onto retentive membranes is now primarily used for immunoblotting. A second application that was quite common up to about a decade ago was electroblotting of proteins for N-terminal and internal sequencing using Edman chemistry. This unit contains procedures for electroblotting proteins from polyacrylamide gels onto a variety of membranes, including polyvinylidene difluoride (PVDF) and nitrocellulose. In addition to the commonly used tank or wet transfer system, protocols are provided for electroblotting using semidry and dry systems. This unit also describes procedures for eluting proteins from membranes using detergents or acidic extraction with organic solvents for specialized applications. PMID:26521711

  8. Nondenaturing electrophoresis of lipoproteins in agarose and polyacrylamide gradient gels

    SciTech Connect

    Shore, V.G.

    1989-12-19

    The plasma lipoproteins frequently are classified according to density and/or electrophoretic mobility. The lipoprotein classes differ characteristically also in particle size and apolipoprotein composition. Each class is heterogeneous in size and composition as well. Nondenaturing electrophoresis in agarose gels and polyacrylamide gradient gels are complementary analytical methods for classification of lipoproteins and determining distribution profiles of the major classes. In addition, gradient gel electrophoresis (GGE) has a high resolving capability for subfractionating each class according to particle size. Combination of gel electrophoresis with immunoblotting yields information on heterogeneity in apolipoprotein distribution. 14 refs., 6 figs., 3 tabs.

  9. Jet injection into polyacrylamide gels: investigation of jet injection mechanics.

    PubMed

    Schramm-Baxter, Joy; Katrencik, Jeffrey; Mitragotri, Samir

    2004-08-01

    Jet injectors employ high-velocity liquid jets that penetrate into human skin and deposit drugs in the dermal or subdermal region. Although jet injectors have been marketed for a number of years, relatively little is known about the interactions of high-speed jets with soft materials such as skin. Using polyacrylamide gels as a model system, the mechanics of jet penetration, including the dependence of jet penetration on mechanical properties, was studied. Jets employed in a typical commercial injector, (orifice diameter: 152 microm, velocity: 170-180 m/s) were used to inject fluid into polyacrylamide gels possessing Young's moduli in the range of 0.06-0.77 MPa and hardness values in the range of 4-70 H(OO). Motion analysis of jet entry into polyacrylamide gels revealed that jet penetration can be divided into three distinct events: erosion, stagnation, and dispersion. During the erosion phase, the jet removed the gel at the impact site and led to the formation of a distinct cylindrical hole. Cessation of erosion induced a period of jet stagnation ( approximately 600 micros) characterized by constant penetration depth. This stage was followed by dispersion of the liquid into the gel. The dispersion took place by crack propagation and was nearly symmetrical with the exception of injections into 10% acrylamide (Young's modulus of 0.06 MPa). The penetration depth of the jets as well as the rate of erosion decreased with increasing Young's modulus. The mechanics of jet penetration into polyacrylamide gels provides an important tool for understanding jet injection into skin.

  10. Use of a bilayer stacking gel to improve resolution of lipopolysaccharides and lipooligosaccharides in polyacrylamide gels.

    PubMed

    Inzana, T J; Apicella, M A

    1999-03-01

    Lipopolysaccharide (LPS) and lipooligosaccharide (LOS) are important antigenic and integral components of the outer membrane of Gram-negative bacteria. Alteration or heterogeneity of LPS/LOS structure is most often assessed by alteration of electrophoretic band profiles using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In order to discern minor differences in the electrophoretic profile of closely spaced bands, particularly the low molecular weight bands of LOS, optimum resolution is required. Unfortunately, many publications of LPS/LOS in polyacrylamide gels show a diffuse, smeared pattern without discernible bands. We report here a formulation for polyacrylamide gels that reproducibly yields LPS/LOS bands with sharp resolution. A key feature of this formulation is the use of a separate comb gel containing electrode buffer layered on top of the conventional stacking gel.

  11. Polyacrylamide gel electrophoresis of intact bacteriophage T4D particles.

    PubMed Central

    Childs, J D; Birnboim, H C

    1975-01-01

    A method for the electrophoresis of intact bacteriophage T4D particles through polyacrylamide gels has been developed. It was found that phage particles will migrate through dilute polyacrylamide gels (less than 2.1%) in the presence of a low concentration of MgCl2. As few as 5 x 10(9) phage particles can be seen directly as a light-scattering band during the course of electrophoresis. The band can also be detected by scanning gels at 260 to 265 nm or by eluting viable phage particles from gel slices. A new mutant (eph1) has been identified on the basis of its decreased electrophoretic mobility compared with that of the wild type; mutant particles migrated 14% slower than the wild type particles at pH 8.3 and 35% slower at pH 5.0. The isoelectric points of both the wild type and eph1 mutant were found to be between pH 4.0 and 5.0. Particles of T4 with different head lengths were also studied. Petite particles (heads 20% shorter than normal) migrated at the same rate as normal-size particles. Giant particles, heterogenous with respect to head length (two to nine times normal), migrated faster than normal-size particles as a diffuse band. This diffuseness was due to separation within the band of particles having mobilities ranging from 8 to 35% faster than those of normal-size particles. These observations extend the useful range of polyacrylamide gel electrophoresis to include much larger particles than have previously been studied, including most viruses. Images PMID:240037

  12. Western Blot of Stained Proteins from Dried Polyacrylamide Gels

    NASA Technical Reports Server (NTRS)

    Gruber, Claudia; Stan-Lotter, Helga

    1996-01-01

    Western blotting of proteins is customarily performed following their separation on polyacrylamide gels, either prior to staining (1) or, as recently reported, following staining (2). We describe here Western blotting with stained gels, which had been dried and some of which had been stored for years. This procedure permits immunological analysis of proteins, to which antisera may have become available only later, or where the application of newly developed sensitive detection methods is desired. Once rehydration of the gels is achieved, proteins can be-transferred to blotting membranes by any appropriate protocol. Proteins stained with Coomassie Blue have to be detected with a non-chromogenic method, such as the film-based enhanced chemiluminescence (ECL)2) procedure (3). Silver stained proteins, which transfer in the colorless form, may be visualized by any detection method, although, because of the usually very low amounts of proteins, detection by ECL is preferable. Blotting of stained proteins from rehydrated gels is as rapid and as quantitative as from freshly prepared gels, in contrast to blotting from wet stained gels, which requires extensive washing and results in low transfer efficiency (2). Together with a photographic record of the gel pattern, unambiguous identification of immunoreactive proteins from complex mixtures is possible. Some further applications of this work are discussed.

  13. High yield electroblotting onto polyvinylidene difluoride membranes from polyacrylamide gels.

    PubMed

    Mozdzanowski, J; Hembach, P; Speicher, D W

    1992-01-01

    Optimal conditions of electroblotting that led to high protein recovery on polyvinylidene difluoride (PVDF) membranes were determined for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). SDS concentrations in the gel and transfer buffer were found to be the most important factors affecting the amount of protein recovered on the PVDF membrane. The largest loss occurred during the first 10-30 min of transfer due to the relatively high initial SDS concentration in the gel. During this initial stage of transfer, most of the protein passed through the primary membrane and was partially retained on secondary and tertiary membranes. The value of presoaking gels prior to transfer to reduce the amount of SDS was evaluated by quantitating free SDS densitometrically and by correlating the reduced SDS concentration with increased electroblotting efficiency from presoaked gels. Transfer time was evaluated and no "overtransfer" was found even after very long transfer times. These results clearly indicate that proteins electroblotted onto PVDF membranes were tightly bound and could not be released by extending the transfer time. The effects of methanol and SDS concentrations on protein adsorption from solution to PVDF were also determined quantitatively. The results of this study strongly suggest that proteins fully saturated with SDS cannot bind efficiently to PVDF membranes. Since SDS is necessary for high protein mobility, the challenge in efficient electroblotting is to maintain an optimal SDS concentration which is high enough to permit effective removal from the gel and low enough to permit effective binding to the PVDF membrane. For 1.5 mm thick gels containing 0.2% SDS, presoaking the gel for 15-20 min in transfer buffer with 10% methanol prior to electroblotting provided the best recovery on the primary membrane.

  14. Scalable lithography from Natural DNA Patterns via polyacrylamide gel

    PubMed Central

    Qu, JieHao; Hou, XianLiang; Fan, WanChao; Xi, GuangHui; Diao, HongYan; Liu, XiangDon

    2015-01-01

    A facile strategy for fabricating scalable stamps has been developed using cross-linked polyacrylamide gel (PAMG) that controllably and precisely shrinks and swells with water content. Aligned patterns of natural DNA molecules were prepared by evaporative self-assembly on a PMMA substrate, and were transferred to unsaturated polyester resin (UPR) to form a negative replica. The negative was used to pattern the linear structures onto the surface of water-swollen PAMG, and the pattern sizes on the PAMG stamp were customized by adjusting the water content of the PAMG. As a result, consistent reproduction of DNA patterns could be achieved with feature sizes that can be controlled over the range of 40%–200% of the original pattern dimensions. This methodology is novel and may pave a new avenue for manufacturing stamp-based functional nanostructures in a simple and cost-effective manner on a large scale. PMID:26639572

  15. Scalable lithography from Natural DNA Patterns via polyacrylamide gel

    NASA Astrophysics Data System (ADS)

    Qu, Jiehao; Hou, Xianliang; Fan, Wanchao; Xi, Guanghui; Diao, Hongyan; Liu, Xiangdon

    2015-12-01

    A facile strategy for fabricating scalable stamps has been developed using cross-linked polyacrylamide gel (PAMG) that controllably and precisely shrinks and swells with water content. Aligned patterns of natural DNA molecules were prepared by evaporative self-assembly on a PMMA substrate, and were transferred to unsaturated polyester resin (UPR) to form a negative replica. The negative was used to pattern the linear structures onto the surface of water-swollen PAMG, and the pattern sizes on the PAMG stamp were customized by adjusting the water content of the PAMG. As a result, consistent reproduction of DNA patterns could be achieved with feature sizes that can be controlled over the range of 40%-200% of the original pattern dimensions. This methodology is novel and may pave a new avenue for manufacturing stamp-based functional nanostructures in a simple and cost-effective manner on a large scale.

  16. Characteristics of polyacrylamide gel with THPC and Turnbull Blue gel dosimeters evaluated using optical tomography

    NASA Astrophysics Data System (ADS)

    Pilařová (Vávrů), Kateřina; Kozubíková, Petra; Šolc, Jaroslav; Spěváček, Václav

    2014-11-01

    The purpose of this study was to compare characteristics of radiochromic gel - Turnbull Blue gel (TB gel) with polymer gel - polyacrylamide gel and tetrakis hydroxymethyl phosphonium chloride (PAGAT) using optical tomography. Both types of gels were examined in terms of dose sensitivity, dose response linearity and background value of spectrophotometric absorbance. The calibration curve was obtained for 60Co irradiation performed on Gammacell 220 at predefined gamma dose levels between 0 and 140 Gy for TBG and 0-15 Gy for PAGAT. To measure relative dose distributions from stereotactic irradiation, dosimeters were irradiated on Leksell Gamma Knife Perfexion. The cylindrical glass housings filled with gel were attached to the stereotactic frame. They were exposed with single shot and 16 mm collimator by 65 Gy to a 50% prescription isodose for TB gel and 4 Gy to a 50% prescription isodose for PAGAT. Evaluations of dosimeters were performed on an UV-vis Spectrophotometer Helios β and an optical cone beam homemade tomography scanner with a 16-bit astronomy CCD camera with a set of color filters. The advantages and potential disadvantages for both types of gel dosimeters were summarized. Dose distribution in central slice and measured profiles of 16 mm shot shows excellent correspondence with treatment planning system Leksell GammaPlan® for both PAGAT and Turnbull Blue gels. Gel dosimeters are suitable for steep dose gradient verification. An optical tomography evaluation method is successful. Dose response characteristics of TB gel and PAGAT gel are presented.

  17. Rheological properties of reactive extrusion modified waxy starch and waxy starch-polyacrylamide copolymer gels

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The rheological properties of modified waxy starch and waxy starch-polyacrylamide graft copolymers prepared by reactive extrusion were investigated. Both materials can absorb huge amount of water and form gels. The modified waxy starch and waxy starch-polyacrylamide graft copolymer gels all exhibite...

  18. Application of SYPRO Ruby- and Flamingo-stained polyacrylamide gels to Western blot analysis.

    PubMed

    Hagiwara, Makoto; Kobayashi, Ken-Ichi; Tadokoro, Tadahiro; Yamamoto, Yuji

    2010-02-15

    Western blots are widely used for analysis of the expression levels of specific proteins. Blotting is conducted after sodium dodecyl sulfate or native polyacrylamide gel electrophoresis without staining the gel. However, when it is necessary to analyze the gel, duplicate polyacrylamide gels (one of which is stained) usually must be prepared, leading to the consumption of precious sample. Thus, we have developed a convenient and efficient Western blot method using a stained gel. This simple modification should be beneficial for the analysis of samples that are limited in quantity and/or samples for which the stained gel serves as the loading control.

  19. Application of SYPRO Ruby- and Flamingo-stained polyacrylamide gels to Western blot analysis.

    PubMed

    Hagiwara, Makoto; Kobayashi, Ken-Ichi; Tadokoro, Tadahiro; Yamamoto, Yuji

    2009-06-15

    Western blot analysis has been a useful method for analysis of expression levels of specific proteins and is conducted after sodium dodecyl sulfate (SDS) or native polyacrylamide gel electrophoresis without staining the gel. However, when it is necessary to analyze the gel, duplicate polyacrylamide gels usually must be prepared, one of which is stained, leading to the consumption of precious sample. Thus, we developed a convenient and efficient Western blotting method using a stained gel. This simple modification should be beneficial for analyzing samples that are limited in quantity and/or samples for which the stained gel serves as the loading control.

  20. Binding of lithium dodecyl sulfate to polyacrylamide gel at 4 degrees C perturbs electrophoresis of proteins.

    PubMed

    Kubo, K; Takagi, T

    1986-07-01

    Although polyacrylamide gel has no affinity to lithium dodecyl sulfate (LDS) at 25 degrees C, the gel maximally binds 17 mg of LDS per gram dry weight at 4 degrees C. When polyacrylamide gel electrophoresis is carried out at 4 degrees C in the presence of LDS instead of sodium dodecyl sulfate (SDS) using a continuous buffer system, migration of proteins with lower molecular weight is accelerated as a result of the deficiency of LDS in the frontal region of the gel. When the gel is saturated with LDS, electrophoresis in the presence of LDS at 4 degrees C shows a resolution higher than that of SDS-polyacrylamide gel electrophoresis at 25 degrees C.

  1. Agarose gel electrophoresis and polyacrylamide gel electrophoresis for visualization of simple sequence repeats.

    PubMed

    Anderson, James; Wright, Drew; Meksem, Khalid

    2013-01-01

    In the modern age of genetic research there is a constant search for ways to improve the efficiency of plant selection. The most recent technology that can result in a highly efficient means of selection and still be done at a low cost is through plant selection directed by simple sequence repeats (SSRs or microsatellites). The molecular markers are used to select for certain desirable plant traits without relying on ambiguous phenotypic data. The best way to detect these is the use of gel electrophoresis. Gel electrophoresis is a common technique in laboratory settings which is used to separate deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) by size. Loading DNA and RNA onto gels allows for visualization of the size of fragments through the separation of DNA and RNA fragments. This is achieved through the use of the charge in the particles. As the fragments separate, they form into distinct bands at set sizes. We describe the ability to visualize SSRs on slab gels of agarose and polyacrylamide gel electrophoresis.

  2. A continuous acetic acid system for polyacrylamide gel electrophoresis of gliadins and other prolamines.

    PubMed

    Clements, R L

    1988-02-01

    A polyacrylamide gel electrophoresis system buffered by acetic acid alone was developed for electrophoresis of prolamines. When applied to gliadin electrophoresis, the acetic acid system produces more bands than does a conventional aluminum lactate-lactic acid system (using 12% acrylamide gels). The acetic acid system is relatively simple, requiring a single buffer component that is universally available in high purity.

  3. Effects of degree of hydrolysis and shear on gelation reaction kinetics and gel strength. [Polyacrylamides

    SciTech Connect

    Gao, Hong W.

    1991-02-01

    Gelation tests were conducted to investigate the effect of the degree of hydrolysis on gelation reaction kinetics and gel strength using four low-molecular-weight polyacrylamides (MW = 400,000 daltons), which were 10% (HPAM1-10), 20% (HPAM1-20), 30% (HAPM1-30), and 40% (HPAM-40) hydrolyzed, and Cr-3 (pH = 4.8) and Al-3 (pH = 7.0) crosslinkers. Results showed that for polymer/Cr-3 gel systems, samples prepared with a low-molecular-weight polyacrylamide polymer, which was 20% hydrolyzed, gelled at a faster rate and retained higher gel strength than those prepared with a low-molecular-weight polyacrylamide polymer, which was 10% hydrolyzed. Under the screening condition, no viscosity enhancement was observed in samples prepared with polymers having a degree of hydrolysis equal to or greater than 30%. For polymer/Al-3 gel systems, samples prepared with a low-molecular-weight polyacrylamide polymer, which was 20% hydrolyzed, gelled at the fastest rate and retained the strongest gel strength among the polymer/Al-3 gel systems prepared with four low-molecular-weight polyacrylamide polymers, which were 10, 20, 30, and 40% hydrolyzed, respectively. Gelation tests of gel systems in glass bead packs showed that high shear favored the gelation of a gel system that had a fast rate of gelation, but had an adverse effect on the gelation of three gel systems that had a slow rate of gelation. Weak gels were found to be injectable through porous media. Weak gels were degradable under high shear condition and regained viscosity under low shear conditions. 17 refs., 8 figs., 1 tab.

  4. Detection of serum proteins by native polyacrylamide gel electrophoresis using Blue Sepharose CL-6B-containing stacking gels.

    PubMed

    Muratsubaki, Haruhiro; Satake, Kaoru; Yamamoto, Yasuhisa; Enomoto, Keiichiro

    2002-08-15

    Analysis of serum proteins by native polyacrylamide gel electrophoresis is difficult because albumin is abundant in serum and interferes with the resolution of other proteins, especially alpha-antitrypsin which has mobility that is very similar to that of albumin. We present here a method in which serum proteins are separated by polyacrylamide gel electrophoresis using stacking gels containing Blue Sepharose CL-6B, which has a high affinity for albumin, lipoproteins, kinases, and pyridine-nucleotide-dependent oxidoreductases. During electrophoresis, proteins that bind to Blue Sepharose CL-6B stay in the stacking gel and do not migrate into the separating gel. As a consequence, certain proteins, including alpha(1)-antitrypsin, can be detected as clear bands. This method overcomes the requirement for fractionation of serum samples prior to electrophoresis to remove albumin and allows the simultaneous analysis of many samples.

  5. Penis invalidating cicatricial outcomes in an enlargement phalloplasty case with polyacrylamide gel (Formacryl).

    PubMed

    Parodi, P C; Dominici, M; Moro, U

    2006-01-01

    The present article reports the case of a patient subjected to polyacrylamide polymers-composed gel cutaneous infiltration in the penis for cosmetic purposes, resulting in severe invalidating outcomes. A significant tissue reaction to the subcutaneous injection of polyacrylamide gel for the penis enlargement purpose resulted in permanent and invalidating scars both on the esthetic and functional levels. Such a result must be simply taken into account both singly and in the light of the international literature to exclude this method as standard uro-andrologic activity.

  6. Biochemical Identification of the Two Races of Radopholus similis by Polyacrylamide Gel Electrophoresis.

    PubMed

    Huettel, R N; Dickson, D W; Kaplan, D T

    1983-07-01

    Analysis of proteins of the banana and citrus race of Radopholus similis was carried out by several different types of polyacrylamide gel electrophoresis. These included standard slab gel, SDS slab gel, gradient slab gel, and two-ditnensional slab gel electrophoresis. A major band difference was detected between the two races by slab gel electrophoresis. However, several other poorly resolved but consistent hands of high molecular weight proteins near the gel origin also were considered as diagnostic. Resolution of protein bands was greatly improved by SDS and gradient slab gel electrophoresis, but no differences could be detected among the proteins resolved between the two rares with these techniques. Two-dimensional gels revealed a large number of proteins, but background staining obscured them hindering interpretation. When nematode races were reared on three different host plants, no differences in protein patterns were detected between them, indicating host preferences does not play a role in determining the types proteins occurring in these nematodes.

  7. Congruence between starch gel and polyacrylamide gel electrophoresis in detecting allozyme variation in pulmonate land slugs.

    PubMed

    Geenen, Sofie; Jordaens, Kurt; Castilho, Rita; Backeljau, Thierry

    2003-02-01

    The predominantly selfing slug species Arion (Carinarion) fasciatus, A. (C.) silvaticus and A. (C.) circumscriptus are native in Europe and have been introduced into North America, where each species consists of a single, homozygous multilocus genotype (strain), as defined by starch gel electrophoresis (SGE) of allozymes. In Europe, the "one strain per species" hypothesis does not hold since polyacrylamide gel electrophoresis (PAGE) of allozymes uncovered 46 strains divided over the three species. However, electrophoretic techniques may differ in their ability to detect allozyme variation. Therefore, several Carinarion populations from both continents were screened by applying the two techniques simultaneously on the same individual slugs and enzyme loci. SGE and PAGE yielded exactly the same results, so that the different degree of variation in North American and European populations cannot be attributed to differences in resolving power between SGE and PAGE. We found four A. (C.) silvaticus strains in North America indicating that in this region the "one strain per species" hypothesis also cannot be maintained. Hence, the discrepancies between previous electrophoretic studies on Carinarion are most likely due to sampling artefacts and possible founder effects.

  8. Two-dimensional polyacrylamide gel analysis of Plodia interpunctella granulosis virus

    SciTech Connect

    Russell, D.L.; Consigli, R.A.

    1986-10-01

    The structural polypeptides of purified Plodia interpunctella granulosis virus were analyzed by three different two-dimensional gel systems. Isoelectric focusing followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis allowed resolution of 53 acidic polypeptides in the enveloped nucleocapsid of the virus ranging in molecular weight from 97,300 to 8000. Nine of these polypeptides were shown to be glycoproteins by the technique of radiolabeled lectin blotting. Separation of the granulin in this system allowed resolution of five species, all of which have identical tryptic peptide maps. This matrix protein was demonstrated to be a phosphoglycoprotein by radiolabeled lectin blotting and acid phosphatase dephosphorylation. Nonequilibrium pH gel electrophoresis followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis allowed resolution of the major basic protein of the virus, VP12, from a more acidic protein of the same molecular weight. Tryptic peptide analysis demonstrated that these two proteins were indeed different and acid urea gels followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis allowed localization of the acidic protein to the envelope and the basic protein to the nucleocapsid of the virus. Finally, probing of the separated envelope nucleocapsid proteins in both the isoelectric focusing and nonequilibrium pH gel electrophoresis two-dimensional systems after transfer to nitrocellulose with iodinated, purified viral proteins allowed further insight into reactions which may be important in the maintenance of the virion structure.

  9. Rheological monitoring of the formation of polyacrylamide/Cr+3 gels

    SciTech Connect

    Prud'Homme, R.K.; Uhl, J.T.; Poinsatte, J.P.; Halverson, F.

    1983-10-01

    The formation of polyacrylamide/chromium-ion gels has been followed rheologically. The time dependence of the storage modulus has been used to analyze the kinetics of the gelation process. The kinetic theory of rubber elasticity has been used to determine the crosslink density in the gel from the measured value of the storage modulus. The effects of changing polymer, chromium ion, and reducing agent concentrations have been studied. 30 references.

  10. One-dimensional SDS-polyacrylamide gel electrophoresis (1D SDS-PAGE).

    PubMed

    Brunelle, Julie L; Green, Rachel

    2014-01-01

    This protocol describes a denaturing polyacrylamide gel system utilizing sodium dodecyl sulfate (SDS) to separate protein molecules based on size as first described by Laemmli (1970). SDS-PAGE can be used to monitor protein purifications, check the purity of samples, and to estimate molecular weights for unknown proteins.

  11. Tris-acetate polyacrylamide gradient gel electrophoresis for the analysis of protein oligomerization.

    PubMed

    Cubillos-Rojas, Monica; Schneider, Taiane; Sánchez-Tena, Susana; Bartrons, Ramon; Ventura, Francesc; Rosa, Jose Luis

    2016-02-01

    Here we report a new approach for studying protein oligomerization in cells using a single electrophoresis gel. We combined the use of a crosslinking reagent for sample preparation, such as glutaraldehyde, with the analysis of oligomers by Tris-acetate polyacrylamide gel electrophoresis. The use of a 3-15% Tris-acetate polyacrylamide gradient gel allows for the simultaneous analysis of proteins of masses ranging from 10 to 500 kDa. We showed the usefulness of this method for analyzing endogenous p53 oligomerization with high resolution and sensitivity in human cells. Oligomerization analysis was dependent on the crosslinker concentration used. We also showed that this method could be used to study the regulation of oligomerization. In all experiments, Tris-acetate polyacrylamide gel electrophoresis proved to be a robust, manageable, and cost- and time-efficient method that provided excellent results using a single gel. This approach can be easily extrapolated to the study of other oligomers. All of these features make this method a highly useful tool for the analysis of protein oligomerization.

  12. Resolution of high molecular weight proteins in dependence on electric field strength in polyacrylamide gel electrophoresis.

    PubMed

    Starita-Geribaldi, M; Houri, A

    1997-01-01

    Resolution of high molecular weight proteins, in the upper region of polyacrylamide gels, was studied in relation to the type of electric field. Separations by constant field gel electrophoresis (CFGE) were compared to those in pulsed oscillatory high-performance electrophoresis (POPE), a novel technique which allows electrophoresis at high field strengths owing to a novel local field distribution. This distribution contributes to structural and mechanical stability of the gel with resultant well-reproducible separation, enhanced resolution, and higher absolute mobility of proteins in POPE.

  13. Polyacrylamide gel electrophoretic methods in the separation of structural muscle proteins.

    SciTech Connect

    Barany, K.; Barany, M.; Giometti, C. S.; Center for Mechanistic Biology and Biotechnology; Univ. of Illinois at Chicago

    1995-04-28

    Polyacrylamide gel electrophoresis plays a major role in analyzing the function of muscle structural proteins. This review describes one- and two-dimensional gel electrophoretic methods for qualitative and quantitative investigation of the muscle proteins, with special emphasis on determination of protein phosphorylation. The electrophoretic studies established the subunit structures of the muscle proteins, characterized their multiple forms, revealed changes in subunit composition or shifts in isoform distribution of specific proteins during development, upon stimulation or denervation of the muscle. Protein phosphorylation during muscle contraction is preferentially studied by two-dimensional gel electrophoresis. The same method demonstrated protein alterations in human neuromuscular diseases.

  14. Enhancement of polyacrylamide gel slice dissolution in hydrogen peroxide by cupric sulfate.

    PubMed

    Donato, H; Doig, M T; Priest, D G

    1988-04-01

    An improved method is described for quantitation of radio-labelled protein by scintillation counting after polyacrylamide gel electrophoresis. The method is based upon copper catalyzed dissolution of gel slices in hydrogen peroxide under ambient conditions. Complete dissolution of gel sections was accomplished by incubation at 25 degrees C in 30% H2O2 that contained 0.9 mM CuSO4. Recovery of tritiated protein was greater than 90% under these conditions while in the absence of CuSO4 recovery was less than 50%.

  15. Renaturation of enzymes after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate

    SciTech Connect

    Lacks, S.A.; Springhorn, S.S.

    1980-08-10

    A number of enzymes, including amylases, dehydrogenases, and proteases, were shown to be renaturable after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Enzyme activity was detected in situ by action on substrates introduced into the gel and subsequent staining of either the product or unreacted substrate. Enzymes appeared to recover activity as soon as the detergent diffused out of the gel. Renatured enzymes were retained in gels after electrophoresis longer than native enzymes which had been subjected to electrophoresis in the absence of detergent. Re-electrophoresis of the renatured enzymes showed that part of the retained activity was physically anchored to the gel, possibly by the folding of polypeptides around the gel matrix as the enzymes were renatured.

  16. Using in situ rheology to characterize the microstructure in photopolymerized polyacrylamide gels for DNA electrophoresis.

    PubMed

    Wang, Jian; Ugaz, Victor M

    2006-09-01

    Photopolymerized cross-linked polyacrylamide hydrogels are attractive sieving matrix formulations for DNA electrophoresis owing to their rapid polymerization times and the potential to locally tailor the gel pore structure through spatial variation of illumination intensity. This capability is especially important in microfluidic systems, where photopolymerization allows gel matrices to be precisely positioned within complex microchannel networks. Separation performance is also directly related to the nanoscale gel pore structure, which is in turn strongly influenced by polymerization kinetics. Unfortunately, detailed studies of the interplay among polymerization kinetics, mechanical properties, and structural morphology are lacking in photopolymerized hydrogel systems. In this paper, we address this issue by performing a series of in situ dynamic small-amplitude oscillatory shear measurements during photopolymerization of cross-linked polyacrylamide electrophoresis gels to investigate the relationship between rheology and parameters associated with the gelation environment including UV intensity, monomer and cross-linker composition, and reaction temperature. In general, we find that the storage modulus G' increases with increasing initial monomer concentration, cross-linker concentration, and polymerization temperature. The steady-state value of G', however, exhibits a more complex dependence on UV intensity that varies with gel concentration. A simple model based on rubber elasticity theory is used to obtain estimates of the average gel pore size that are in surprisingly good agreement with corresponding data obtained from analysis of DNA electrophoretic mobility in gels cast under identical polymerization conditions.

  17. [THE USE OF THE COMMERCIAL APPARATUS DUAL GEL MODULE FOR THE TWO-DIMENSIONAL POLYACRYLAMIDE GEL ELECTROPHORESIS].

    PubMed

    Evteeva, I N; Starkova, T Yu; Artemov, A V; Barlev, N A

    2015-01-01

    Two-dimensional gel electrophoresis, continues to be one of the fundamental methods to study the biological protein diversity. This method described by O'Farrell in 1975 includes two following steps: isoelectric focusing in the first dimension and polyacrylamide gel electrophoretic fractionation of proteins according to their molecular weight in the second dimension. In this manuscript we described several technical parameters of the commercial apparatus Dual Gel Module for the gel electrophoresis by means of which it is possible to accomplish the electrophoretic protein fractionation in both dimensions. The distribution of the highly purified commercial proteins used as molecular standards in the detection system of the apparatus Dual Gel Module was identical to the commercial strips of the device GE Healthcare, USA.

  18. Detection of connexins in liver cells using sodiumdodecylsulfate polyacrylamide gel electrophoresis and immunoblot analysis

    PubMed Central

    Willebrords, Joost; Maes, Michaël; Yanguas, Sara Crespo; Cogliati, Bruno; Vinken, Mathieu

    2016-01-01

    Summary Since connexin expression is partly regulated at the protein level, immunoblot analysis represents a frequently addressed technique in the connexin research field. The present chapter describes the set-up of an immunoblot procedure, including protein extraction and quantification from biological samples, gel electrophoresis, protein transfer and immunoblotting, which is optimized for analysis of connexins in liver tissue. In essence, proteins are separated on a polyacrylamide gel using sodiumdodecylsulfate followed by transfer of proteins on a nitrocellulose membrane. The latter allows specific detection of connexins with antibodies combined with revelation through enhanced chemiluminescence. PMID:27207285

  19. Highly sensitive fluorescent stain for detecting lipopolysaccharides in sodium dodecyl sulfate polyacrylamide gel electrophoresis.

    PubMed

    Wang, Xu; Zhou, Ayi; Cai, Wanhui; Yu, Dongdong; Zhu, Zhongxin; Jiang, Chengxi; Jin, Litai

    2015-08-01

    A sensitive and simple technique was developed for the visualization of gel-separated lipopolysaccharides by using a hydrazide derivative, UGF202. As low as 0.5-1 ng total LPS could be detected by UGF202 stain, which is 2- and 16-fold more sensitive than that of the commonly used Pro-Q Emerald 300 and Keenan et al. developed silver stain, respectively. The results indicated that UGF202 stain could be a good choice for LPS determination in polyacrylamide gels.

  20. 'Catalysts' for polyacrylamide gel polymerization and detection of proteins by silver staining.

    PubMed

    Hochstrasser, D F; Merril, C R

    1988-01-01

    The crosslinker diacrylyl-piperazine produces polyacrylamide gels which display improved electrophoretic separation of proteins and better physical strength. It also produces gels with improved detection of proteins by ammoniacal silver staining by reducing the background. This reduced background provided us with an opportunity to investigate residual background staining caused by the catalytic reagents utilized in the polymerization of acrylamide gels. The commonly used catalyst system, tetramethyl-ethylenediamine and ammonium persulfate was shown to be responsible for the yellow staining background found after a prolonged development time with silver staining. An alternate catalyst system has been designed to decrease further the formation of this background staining. Dimethyl-piperazine or tetramethylethylenediamine, potassium or ammonium persulfate, and sodium thiosulfate are shown to provide for gels which have excellent mechanical and staining characteristics. These catalytic systems produce little background staining despite prolonged development time with the ammoniacal silver stain, and they reduce background staining with the dichromate silver stain.

  1. Peptide fractionation by SDS-free polyacrylamide gel electrophoresis for proteomic analysis via DF-PAGE.

    PubMed

    Ramos, Yassel; Besada, Vladimir; Castellanos-Serra, Lila

    2012-01-01

    Here we present a procedure for peptide fractionation by SDS-free polyacrylamide gel electrophoresis, based on discontinuous buffer systems. In the absence of SDS, peptide migration depends both on their molecular mass and on their net charge at the electrophoresis pH. By selecting the separation pH, peptide mobility is modulated. In the original discontinuous buffer system (Tris/glycine), peptides that migrate to the anode have pI values below 6.8 and distribute along the lane in a pI decreasing order, while at acidic pH, as that afforded by histidine/MOPS buffer system, peptides with pI below 5.5 are fractionated. Separation at acid pH is particularly useful for recovering phosphopeptides as well as other highly negatively charged peptides, as those containing sialic or sulfate substituents. Both separation conditions in Tris/glycine and in histidine/MOPS are applicable to proteomic studies, by dual-fractionation polyacrylamide gel electrophoresis (DF-PAGE). First, complex protein samples are separated via SDS-PAGE, and after in-gel proteolysis, peptides are loaded on a second SDS-free gel, where they are separated as described here.

  2. Glutamine Synthetase Regulation, Adenylylation State, and Strain Specificity Analyzed by Polyacrylamide Gel Electrophoresis

    PubMed Central

    Bender, Robert A.; Streicher, Stanley L.

    1979-01-01

    We used polyacrylamide gel electrophoresis to examine the regulation and adenylylation states of glutamine synthetases (GSs) from Escherichia coli (GSE) and Klebsiella aerogenes (GSK). In gels containing sodium dodecyl sulfate (SDS), we found that GSK had a mobility which differed significantly from that of GSE. In addition, for both GSK and GSE, adenylylated subunits (GSK-adenosine 5′-monophosphate [AMP] and GSE-AMP) had lesser mobilities in SDS gels than did the corresponding non-adenylylated subunits. The order of mobilities was GSK-AMP < GSK < GSE-AMP < GSE. We were able to detect these mobility differences with purified and partially purified preparations of GS, crude cell extracts, and whole cell lysates. SDS gel electrophoresis thus provided a means of estimating the adenylylation state and the quantity of GS present independent of enzymatic activity measurements and of determining the strain origin. Using SDS gels, we showed that: (i) the constitutively produced GS in strains carrying the glnA4 allele was mostly adenylylated, (ii) the GS-like polypeptide produced by strains carrying the glnA51 allele was indistinguishable from wild-type GSK, and (iii) strains carrying the glnA10 allele contained no polypeptide having the mobility of GSK or GSK-AMP. Using native polyacrylamide gels, we detected the increased amount of dodecameric GS present in cells grown under nitrogen limitation compared with cells grown under conditions of nitrogen excess. In native gels there was neither a significant difference in the mobilities of adenylylated and non-adenylylated GSs nor a GS-like protein in cells carrying the glnA10 allele. Images PMID:33958

  3. Quantitation of specific proteins in polyacrylamide gels by the elution of Fast Green FCF.

    PubMed

    Gilmore, L B; Hook, G E

    1981-07-01

    The quantitation of proteins in polyacrylamide gels stained with Fast green FCF has been investigated using a modification of the elution technique originally described by Fenner et al. (Fenner, C., Traut, R.R., Mason, D.T. and Wikman-Coffelt, J. (1975) Anal. Biochem. 63, 595--602) for Coomassie Blue and adapted by Medugorac (Medugorac, I. (1979) Basic Res. Cardiol. 74, 406--416) for use with proteins stained with Fast Green FCF. The elution of dye from stained protein was accomplished using 1.0 M NaOH instead of aqueous pyridine as required by the original method. The primary advantages of our modification are that the time required for protein quantitation has been considerably reduced and the use of toxic organic solvents has been eliminated. We have investigated the applicability of the method of several different proteins and our results indicate: (a) The quantity of Fast Green FCF eluted from specific proteins is proportional to the quantity of protein applied to the gel, but varies for each individual protein. (b) The method allows quantitation over a very wide range of protein (1--800 micrograms). (c) Quantitation of protein is independent of the width of the stained bands as well as acrylamide concentration. (d) The method is applicable to gels of many types including disc, slab and continuous gradient gels. (e) Protein can be estimated from the patterns obtained by two-dimensional polyacrylamide gel electrophoresis. (f) The presence of Triton X-100 in gel and protein sample does not affect quantitation; the method is applicable to gels containing SDS provided that SDS is removed prior to staining. (g) Precipitation of protein with 12.5% TCA following electrophoresis does not interfere with quantitation. (h) The reproducibility of the technique is excellent, with standard deviations being less than 10% of the mean in all cases. This method appears highly versatile but requires appropriate standards for the quantitation of individual proteins.

  4. Analysis of mucosal mucins separated by SDS-urea agarose polyacrylamide composite gel electrophoresis.

    PubMed

    Issa, Samah M A; Schulz, Benjamin L; Packer, Nicolle H; Karlsson, Niclas G

    2011-12-01

    Efficient separation of mucins (200 kDa-2 MDa) was demonstrated using gradient SDS agarose/polyacrylamide composite gel electrophoresis (SDS-AgPAGE). Inclusion of urea (SDS-UAgPAGE) in the gels casting were shown to have no effect on the migration of mucins in the gel and allowed casting of gel at room temperature. This simplified the procedure for multiple casting of agarose polyacrylamide gradients and increased reproducibility of these gels. Hence, the implementation of urea makes the technique applicable for high throughput isolation and screening of mucin oligosaccharides by LC-MS after releasing the oligosaccharides from isolated, blotted mucin subpopulations. It was also shown that the urea addition had no effect on other supporting applications such as western and lectin blotting. In addition, identification of the mucin protein after tryptic digestion and LC-MS was possible and no protein carbamylation due to the presence of urea in the gel was detected. LC-MS software developed for metabolomic analysis was used for O-linked oligosaccharide detection and differential display of various mucin samples. Using this method, heterogeneous glycosylation of mucins and mucin-type molecules isolated by SDS-AgPAGE and SDS-UAgPAGE was shown to consist of more than 80 different components in a single band, and in the extreme cases, up to 300-500 components (MUC5B/AC from saliva and sputum and). Metabolomic software was also used to show that the migration of mucin isoforms within the gel is due to heterogeneous size distribution of the oligosaccharides, with the slower migrating bands enriched in high-molecular-weight oligosaccharides.

  5. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography.

    PubMed

    Laskey, R A; Mills, A D

    1975-08-15

    Methods which use the scintillator PPO to record film images of 3H in chromatograms and polyacrylamide gels (fluorography) have been described elsewhere. This paper demonstrates that pre-exposure of the film to a brief flash of light greatly increases the sensitivity of fluorography. Pre-exposure also permits quantitative interpretation of the film image, because it corrects the non-linear relationship between radioactivity of the sample and absorbance of the film image. Therefore the distribution of radioactivity in the sample is accurately represented by microdensitometry of the image obtained on pre-exposed film. Using pre-exposed film 300 dis. 3H/min or 30 dis. 14C/min can be detected in a band in a gel in a 24-h exposure. The Appendix describes revisions and extensions of existing fluorographic procedures, including application to agarose gels and a rapid procedure for recovering PPO for re-use.

  6. Shotgun electroelution: a proteomic tool for simultaneous sample elution from whole SDS-polyacrylamide gel slabs.

    PubMed

    Antal, József; Bányász, Borbála; Buzás, Zsuzsanna

    2007-02-01

    A high-throughput device has been constructed which allows parallel electroelution of separated SDS-protein bands directly from intact unsectioned polyacrylamide gel slabs as well as single electroelution of certain protein spots into a 384-well standard flat-bottom multiwell plate. The prototype provides complete, quick elution for proteomics from 1-D or from 2-D gels without gel sectioning. Since the elution chamber matrix requires no assembly, sample handling can be easily carried out by existing robotic workstations. The current design is a good candidate for automation of spot elution since there are no moving liquid containing components in the apparatus. Eight SDS-proteins were eluted in test runs and an average 70% sample recovery was achieved by re-electrophoresis of the electro-eluates.

  7. Accommodating brightness and exposure levels in densitometry of stained polyacrylamide electrophoresis gels

    SciTech Connect

    Tan, Han Yen; Ng, Tuck Wah; Liew, Oi Wah

    2010-03-20

    Flatbed scanner densitometers can be operated under various illumination and recording exposure levels. In this work, we show that optical density measurement accuracy, sensitivity, and stability of stained polyacrylamide electrophoresis gel densitometry are crucially dependent on these two factors (brightness and exposure level), notwithstanding that the source is monochromatic, spatially uniform, and the measurements are made using an accurately calibrated step wedge in tandem. We further outline a method to accommodate the intensity deviations over a range of illumination and exposure levels in order to maintain sensitivity and repeatability in the computed optical densities. Comparisons were also made with results from a commercial densitometer.

  8. Electroelution of nucleic acids from polyacrylamide gels: a custom-made, agarose-based electroeluter.

    PubMed

    Fadouloglou, Vasiliki E

    2013-06-01

    Polyacrylamide electrophoresis is routinely used for small-scale preparative and analytical separations. The incomparably high-resolution separations achieved by this technique, however, have not been widely exploited to the large-scale preparative isolation of biological molecules from contaminants, mainly because of difficulties in the recovery of the desired molecule from the gel matrix. Electroelution is an effective procedure applied for this purpose. However, commercially available, high-cost electroeluters are required for achieving high recovery yields. Here, we describe a custom-made electroeluter that combines low-cost, high-recovery yields, short times of electroelution, and convenience in the manipulation of sensitive samples.

  9. Electrostatic protein immobilization using charged polyacrylamide gels and cationic detergent microfluidic Western blotting.

    PubMed

    Kim, Dohyun; Karns, Kelly; Tia, Samuel Q; He, Mei; Herr, Amy E

    2012-03-06

    We report a novel protein immobilization matrix for fully integrated microfluidic Western blotting (WB). The electrostatic immobilization gel (EIG) enables immobilization of all proteins sized using cetyl trimethylammonium bromide polyacrylamide gel electrophoresis (CTAB-PAGE), for subsequent electrophoretic probing with detection affinity reagents (e.g., labeled antibodies). The "pan-analyte" capture strategy introduced here uses polyacrylamide gel grafted with concentrated point charges (zwitterionic macromolecules), in contrast to existing microfluidic WB strategies that rely on a sandwich immunoassay format for analyte immobilization and detection. Sandwich approaches limit analyte immobilization to capture of only a priori known targets. A charge interaction mechanism study supports the hypothesis that electrostatic interaction plays a major role in analyte immobilization on the EIG. We note that protein capture efficiency depends on both the concentration of copolymerized charges and ionic strength of the gel buffer. We demonstrate pan-analyte immobilization of sized CTAB-laden model proteins (protein G, ovalbumin, bovine serum albumin, β-galactosidase, lactoferrin) on the EIG with initial capture efficiencies ranging from 21 to 100%. Target proteins fixed on the EIG (protein G, lactoferrin) are detected using antibody probes with signal-to-noise ratios of 34 to 275. The approach advances protein immunoblotting performance through 200× reduction on sample consumption, 12× reduction in assay duration, and automated assay operation, compared to slab-gel WB. Using the microfluidic WB assay, assessment of lactoferrin in human tear fluid is demonstrated with a goal of advancing toward nonbiopsy-based diagnosis of Sjögren's Syndrome, an autoimmune disease.

  10. Tris-acetate polyacrylamide gradient gels for the simultaneous electrophoretic analysis of proteins of very high and low molecular mass.

    PubMed

    Cubillos-Rojas, Monica; Amair-Pinedo, Fabiola; Tato, Irantzu; Bartrons, Ramon; Ventura, Francesc; Rosa, Jose Luis

    2012-01-01

    Polyacrylamide gel electrophoresis (PAGE) is one of the most powerful tools used for protein analysis. We describe the use of Tris-acetate buffer and 3-15% polyacrylamide gradient gels to simultaneously separate proteins in the mass range of 10-500 kDa. We show that this system is highly sensitive, it has good resolution and high reproducibility, and that it can be used for general applications of PAGE such as Coomassie Brilliant Blue staining and immunoblotting. Moreover, we describe how to generate mini Tris-acetate polyacrylamide gels to use them in miniprotein electrophoresis systems. These economical gels are easy to generate and to manipulate and allow a rapid analysis of proteins. All these features make the Tris-acetate-PAGE system a very helpful tool for protein analysis.

  11. Analysis of mitochondrial respiratory chain supercomplexes using blue native polyacrylamide gel electrophoresis (BN-PAGE)

    PubMed Central

    Jha, Pooja; Wang, Xu; Auwerx, Johan

    2016-01-01

    Mitochondria are cellular organelles that produce energy in the form of ATP through a process termed oxidative phosphorylation (OXPHOS), which occurs via the protein complexes of the electron transport chain (ETC). In recent years it has become unequivocally clear that mitochondrial complexes of the ETC are not static entities in the inner mitochondrial membrane. These complexes are dynamic and in mammals they aggregate in different stoichiometric combinations to form supercomplexes (SCs) or respirasomes. It has been proposed that the net respiration is more efficient via SCs than via isolated complexes. However, it still needs to be determined whether the activity of a particular SC is associated with a disease etiology. Here we describe a simplified method to visualize and assess in-gel activity of SCs and the individual complexes with a good resolution on blue native polyacrylamide gel electrophoresis (BN-PAGE). PMID:26928661

  12. Improved detection of amylase activity by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with copolymerized starch.

    PubMed

    Martínez, T F; Alarcón, F J; Díaz-López, M; Moyano, F J

    2000-08-01

    An improved method, based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for detection of amylase activity is described. This method will allow better characterization of certain amylases than that obtained by the Davis technique. The main features of the technique are: (i) identification of amylase bands and molecular mass determination are possible in the same gel; (ii) the hydrolysis of copolymerized substrate during electrophoretic separation is prevented using very low temperatures instead of inactivating agents such as chelating agents; and (iii) the technique is applicable to reveal amylase activity in a wide range of biological samples. The method is not useful for enzymes sensitive to SDS and for high molecular mass amylases.

  13. Microfluidic polyacrylamide gel electrophoresis with in situ immunoblotting for native protein analysis.

    PubMed

    He, Mei; Herr, Amy E

    2009-10-01

    We introduce an automated immunoblotting method that reports protein electrophoretic mobility and identity in a single streamlined microfluidic assay. Native polyacrylamide gel electrophoresis (PAGE) was integrated with subsequent in situ immunoblotting. Integration of three PA gel elements into a glass microfluidic chip achieved multiple functions, including (1) rapid protein separation via on-chip PAGE, (2) directed electrophoretic transfer of resolved protein peaks to an in-line blotting membrane, and (3) high-efficiency identification of the transferred proteins using antibody-functionalized blotting membranes. In-chip blotting membranes were photopatterned with biotinylated antibody using streptavidin polyacrylamide (PA) thus yielding postseparation sample analysis. No pressure driven flow or fluid valving was required, as the assay was operated by electrokinetically programmed control. A model sample of fluorescently labeled BSA (negative control), alpha-actinin, and prostate specific antigen (PSA) was selected to develop and characterize the assay. A 5 min assay time was required without operator intervention. Optimization of the blotting membrane (geometry, operation, and composition) yielded a detection limit of approximately 0.05 pg (alpha-actinin peak). An important additional blotting fabrication strategy was developed and characterized to allow vanishingly small antibody consumption (approximately 1 microg), as well as end-user customization of the blotting membrane after device fabrication and storage. This first report of rapid on-chip protein PAGE integrated with in situ immunoblotting forms the basis for a sensitive, automated approach applicable to numerous forms of immunoblotting.

  14. Microviscosity in polyacrylamide gels with pendant triphenyl-methane leuco derivatives: picosecond time-resolved fluorescence study

    NASA Astrophysics Data System (ADS)

    Tamai, Naoto; Ishikawa, Masazumi; Kitamura, Noboru; Masuhara, Hiroshi

    1991-10-01

    Picosecond fluorescence dynamics of triphenylmethane dyes bonded to polyacrylamide gels before and after swelling was studied by a single-photon timing technique. Microviscosity in the gels after swelling was estimated to be 10-11 cP by examining the viscosity dependence of fluorescence dynamics of malachite green in various alcohols. The results were interpreted in terms of structured stiff water in a microcavity of the gels.

  15. Immobilization of urease from pigeonpea (Cajanus cajan L.) in polyacrylamide gels and calcium alginate beads.

    PubMed

    Das, N; Kayastha, A M; Malhotra, O P

    1998-02-01

    Urease from pigeonpea was entrapped in polyacrylamide gel with 50% immobilization at 10% total monomer (containing 5% cross-linker) with high mechanical stability of the gel. Approximately 0.61 mg of protein could be loaded per 5 ml of gel. The immobilized enzyme had a t1/2 of approx. 200 days when stored in 0.1 M Tris/acetate buffer, pH 6.5, at 4 degrees C. The gel strips were used 4-5 times for urea assay over a period of 6 h with less than 2% loss of activity. Approximately 50% immobilization of urease in calcium alginate was observed at 3% alginate with 0.12 mg protein/ml alginate. The resultant enzyme beads showed a t1/2 of approx. 75 days when stored in 0.1 M Tris/acetate buffer, pH 6.5, at 4 degrees C. The beads were used 4-5 times for urea assay over a period of 6 h with about 40% loss of activity. In both cases, the enzyme activity was directly proportional to the amount of immobilized enzyme. There was practically no leaching of the entrapped enzyme over a period of 48 h from either of the polymers. Both the immobilized enzyme preparations were used to analyse the blood urea of some clinical samples from the University hospital. The results obtained compared favourably with those obtained by the usual method employed in the clinical pathology laboratory.

  16. Highly increased detection of silver stained protein bands in polyacrylamide gels with thermo-optical methods

    NASA Astrophysics Data System (ADS)

    Mazza, Giulia; Posnicek, Thomas; Brandl, Martin

    2016-11-01

    Sodium dodecyl sulfate polyacrylamide gel electrophoresis is a well-known technique to separate proteins by their molecular weight. After electrophoresis, the gels are commonly stained for protein band analysis with silver stain; this allows the detection of protein loads to about 1 ng. To increase the detection sensitivity of the protein bands down in the subnanogram level, a sensor has been developed based on the thermal lens effect to scan and quantify protein loads which would remain undetected using the standard imaging systems. The thermal lens sensor is equipped with a 450 nm diode pump laser modulated at 1 Hz and a HeNe probe laser mounted in collinear geometry. The sensor could detect protein bands of 0.05 ng when the gel was soaked in methanol/water and 0.1 ng in water. The limit of detection ranged from 8 to 20 pg, depending on the soaking medium and the staining efficiency. Thus, the detection of silver stain by thermal lens effect results 10 to 20 times more sensitive than the standard colorimetric method.

  17. Comparison of fluorographic methods for detecting radioactivity in polyacrylamide gels or on nitrocellulose filters

    SciTech Connect

    Roberts, P.L.

    1985-06-01

    The commercial fluorographic enhancers, En3Hance or Amplify, were not as efficient as 2,5-diphenyloxazole (PPO) for detecting radioactively labeled proteins in polyacrylamide gels or on nitrocellulose filters. For most of the X-ray films tested, optimal preexposure was essential to obtain maximum sensitivity in fluorography or indirect autoradiography using intensifying screens. The best results were obtained with nitrocellulose by saturating the filters with PPO. The minimum levels of /sup 35/S//sup 14/C that could be detected on filters by autoradiography or fluorography in a 24-h exposure were 4 X 10(2) or 1 X 10(2) dpm cm-2 respectively. For /sup 3/H these levels were, respectively, 20 X 10(3) or 0.5 X 10(3) dpm cm-2.

  18. Performing isoelectric focusing and simultaneous fractionation of proteins on a rotary valve followed by sodium dodecyl-polyacrylamide gel electrophoresis.

    PubMed

    Wang, Wei; Lu, Joann J; Gu, Congying; Zhou, Lei; Liu, Shaorong

    2013-07-16

    In this technical note, we design and fabricate a novel rotary valve and demonstrate its feasibility for performing isoelectric focusing and simultaneous fractionation of proteins, followed by sodium dodecyl-polyacrylamide gel electrophoresis. The valve has two positions. In one position, the valve routes a series of capillary loops together into a single capillary tube where capillary isoelectric focusing (CIEF) is performed. By switching the valve to another position, the CIEF-resolved proteins in all capillary loops are isolated simultaneously, and samples in the loops are removed and collected in vials. After the collected samples are briefly processed, they are separated via sodium dodecyl-polyacrylamide gel electrophoresis (SDS-PAGE, the second-D separation) on either a capillary gel electrophoresis instrument or a slab-gel system. The detailed valve configuration is illustrated, and the experimental conditions and operation protocols are discussed.

  19. Electrophoresis of /sup 35/S-labeled proteoglycans of polyacrylamide-agarose composite gels and their visualization by fluorography

    SciTech Connect

    Carney, S.L.; Bayliss, M.T.; Collier, J.M.; Muir, H.

    1986-01-01

    Techniques for the electrophoresis of /sup 35/S-labeled proteoglycans on polyacrylamide-agarose gel slabs and subsequent fixation, impregnation, and fluorography of such electrophoretograms have been developed. The procedure permits the examination of newly synthesized proteoglycan subspecies using a rapid technique, previously unavailable for these labeled molecules.

  20. Studies on the bioactivity of radioiodinated highly purified bovine thyrotropin: analytical polyacrylamide gel electrophoresis

    SciTech Connect

    Takai, N.A.; Filetti, S.; Rapoport, B.

    1981-01-01

    Highly purified bovine TSH (stored in solution at -70 C) was radioiodinated by the stoichiometric chloroamine-T method. The iodinated material ws subjected to analytical polyacrylamide disc gel electrophoresis. TSH was eluted from gel slices (1 mm width) and was analyzed for radioactivity and bioactivity. The latter was determined using the cultured thyroid cell cAMP response assay. Radioactivity in the TSH preparation migrated separately from bioactivity, but concordant with the protein bands observed in gels run in parallel. Further studies performed on bovine TSH purified in our laboratory, as well as on a different TSH preparation of exceptionally high potency (both stored as lyophilized powder) revealed a different pattern, with TSH bioactivity and radioactivity eluting concurrently. Iodination of TSH did not alter its electrophoretic migration on disc gel electrophoresis. In all preparations polymorphism of TSH bioactivity was observed, with at least four separate protein bands containing TSH bioactivity being present in our preparation. The relationship between the degree of iodination and retention of TSH bioactivity was examined. Incorporation of /sup 125/I into TSH was greatly different at two different concentrations of chloramine-T. Despite this, however, the progressive loss of TSH bioactivity was similar at both concentrations, indicating that incorporation of iodine into the TSH molecule is not itself responsible for the decrease in bioactivity. These studies indicate variability among different TSH preparations in terms of their retention of bioactivity. Significant loss of TSH bioactivity appears to occur during storage in solution. The damage to the biological activity of TSH during the iodination procedure is more likely related to the oxidation process than to the incorporation of iodine.

  1. Carbon nanotube-modified sodium dodecyl sulfate-polyacrylamide gel electrophoresis for molecular weight determination of proteins.

    PubMed

    Parthasarathy, Meera; Debgupta, Joyashish; Kakade, Bhalchandra; Ansary, Abu A; Islam Khan, M; Pillai, Vijayamohanan K

    2011-02-15

    The effect of incorporating carbon nanotubes (CNTs) in the gel matrix on the electrophoretic mobility of proteins based on their molecular weight differences was investigated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). More specifically, a reduction in standard deviation in the molecular weight calibration plots by 55% in the case of multiwalled carbon nanotubes (MWCNTs) and by 34% in the case of single-walled carbon nanotubes (SWCNTs) compared with that of pristine polyacrylamide gels was achieved after incorporating an insignificant amount of functionalized CNTs into the gel matrix. A mechanism based on a more uniform pore size distribution in CNT modified polyacrylamide gel matrix is proposed. Furthermore, the impact of SWCNTs and MWCNTs on the mobility of proteins in different molecular weight regimes at a given acrylamide concentration offers a tunable gel matrix in terms of the selection of molecular weight ranges of proteins. The robustness and excellent reproducibility of the CNT-PAGE protocol are expected to have a significant impact on the molecular weight determination of newly isolated proteins.

  2. Serum alkaline phosphatase isoenzymes in laboratory beagle dogs detected by polyacrylamide-gel disk electrophoresis.

    PubMed

    Hatayama, Kazuhisa; Nishihara, Yoshito; Kimura, Sayaka; Goto, Ken; Nakamura, Daichi; Wakita, Atsushi; Urasoko, Yoshinaka

    2011-10-01

    Serum alkaline phosphatase (ALP) activity is frequently measured in toxicity studies. Itoh et al. (2002) reported that a commercially available polyacrylamide-gel (PAG) disk electrophoresis kit used in humans (AlkPhor System, Jokoh Co., Ltd., Tokyo, Japan) for identifying serum ALP isoenzymes was useful for veterinary clinicopathological diagnosis in mongrel dogs. In the present study, based on the report of Itoh et al. (2002), we tried to expand the application range of this kit to laboratory beagle dogs which are commonly used in toxicity studies. In order to identify the origin of each ALP isoenzyme, tissue ALP extracts from the liver, bone and small intestine and serum samples were treated with neuraminidase, anti-small intestinal ALP antibody, ALP inhibitor levamisole and/or wheat germ agglutinin (WGA). The main serum ALP isoenzymes in 5-month-old intact beagle dogs were bone-derived (bone and atypical ALP: corresponding to human variant bone ALP) and they tended to decrease with age. However, liver-derived ALP isoenzyme greatly increased in the serum of cholestasis model dogs. The cholestasis model dogs also had a large molecular ALP detected in the resolving gel. This ALP could be originated from intestinal ALP or corticosteroid-induced ALP (CALP), because the activity remained even after levamisole inhibition. CALP was observed in intact laboratory beagle dogs with individual differences. These results suggest that the present method is a useful tool for detecting serum ALP isoenzymes in laboratory beagle dogs and concomitant levamisole inhibition with another gel is applicable for the evaluation of organ toxicity.

  3. Agarose and polyacrylamide gel electrophoresis methods for molecular mass analysis of 5- to 500-kDa hyaluronan.

    PubMed

    Bhilocha, Shardul; Amin, Ripal; Pandya, Monika; Yuan, Han; Tank, Mihir; LoBello, Jaclyn; Shytuhina, Anastasia; Wang, Wenlan; Wisniewski, Hans-Georg; de la Motte, Carol; Cowman, Mary K

    2011-10-01

    Agarose and polyacrylamide gel electrophoresis systems for the molecular mass-dependent separation of hyaluronan (HA) in the size range of approximately 5-500 kDa were investigated. For agarose-based systems, the suitability of different agarose types, agarose concentrations, and buffer systems was determined. Using chemoenzymatically synthesized HA standards of low polydispersity, the molecular mass range was determined for each gel composition over which the relationship between HA mobility and logarithm of the molecular mass was linear. Excellent linear calibration was obtained for HA molecular mass as low as approximately 9 kDa in agarose gels. For higher resolution separation, and for extension to molecular masses as low as approximately 5 kDa, gradient polyacrylamide gels were superior. Densitometric scanning of stained gels allowed analysis of the range of molecular masses present in a sample as well as calculation of weight-average and number-average values. The methods were validated for polydisperse HA samples with viscosity-average molecular masses of 112, 59, 37, and 22 kDa at sample loads of 0.5 μg (for polyacrylamide) to 2.5 μg (for agarose). Use of the methods for electrophoretic mobility shift assays was demonstrated for binding of the HA-binding region of aggrecan (recombinant human aggrecan G1-IGD-G2 domains) to a 150-kDa HA standard.

  4. Carbon nanotubes-assisted polyacrylamide gel electrophoresis for enhanced separation of human serum proteins and application in liverish diagnosis.

    PubMed

    Jiang, Fubin; Wang, Yanan; Hu, Xinfang; Shao, Na; Na, Na; Delanghe, Joris R; Ouyang, Jin

    2010-11-01

    The application of pore-gradient polyacrylamide gel electrophoresis (PG-PAGE) incorporated with carbon nanotube modified by Triton X-100 and carboxylation so as to improve the separation of human serum proteins is reported. The novel PG-PAGE was made by adding water-soluble single-walled carbon nanotubes (CNTs) when preparing the polyacrylamide gel. Significant improvements in separation of complement C3 protein and haptoglobin (Hp) in human serum were achieved. It was estimated that the interactions between the hydrophilic groups on the proteins and the surface of the CNTs result in different adsorption kinetics of complement C3 and Hp subtype on the nanoparticles incorporated in the gel, thus enhancing the separation of the two proteins in serum. This new CNT matrix-assisted PG-PAGE method for enhanced separation of complement C3 and Hp in human serum was successfully applied to distinguish the samples from liverish patients and healthy people.

  5. A shortcut organic dye-based staining method for the detection of DNA both in agarose and polyacrylamide gel electrophoresis.

    PubMed

    Cong, Weitao; Chen, Mao; Zhu, Zhongxin; Liu, Zhiguo; Nan, Jia; Ye, Weijian; Ni, Maowei; Zhao, Ting; Jin, Litai

    2013-02-21

    In this study, we describe a brief, sensitive and safe organic dye-based staining method for the visualization of DNA both in agarose and polyacrylamide gels by using Victoria Pure Blue BO (VPBBO). Down to 0.8-1.6 ng of λ DNA/HindIII markers in agarose gels and 0.4-0.8 ng of pUC18 DNA/Mspl markers in polyacrylamide gels can be successfully detected within 15 and 10 min by the new developed technique, respectively. Moreover, the mechanism of the VPBBO staining was investigated and further confirmed by electrospray ionization mass spectrometry (ESI-MS) and molecular docking. The results indicated that the interaction between VPBBO and DNA is mainly due to groove binding.

  6. Pulsed field electrophoresis for the separation of protein-sodium dodecyl sulfate-complexes in polyacrylamide gels.

    PubMed

    Houri, A; Starita-Geribaldi, M

    1994-01-01

    Polyacrylamide gel electrophoresis of proteins was studied using a pulsed-current mode. A new "local field" distribution was used to correct the gel patterns and optimize migration. A corrective field was applied at fixed 2 s intervals to a constant field, inducing a complex relaxation mechanism. Calculated variations in the local field directions decreased the electric strain on the gel during the run, with resultant optimum gel structure. The relaxation mechanism was found to enhance the absolute mobility of proteins with shorter running times compared to constant field gel electrophoresis (CFGE) and other pulsed field techniques. The enhancement of molecular mobility was explored by transverse pore gradient gel electrophoresis. Ferguson curves which exhibited a convex shape in CFGE were linearized by the new pulsed-field method named pulsed oscillatory high-performance electrophoresis (POPE).

  7. Monthly variations in ovine seminal plasma proteins analyzed by two-dimensional polyacrylamide gel electrophoresis.

    PubMed

    Cardozo, J A; Fernández-Juan, M; Forcada, F; Abecia, A; Muiño-Blanco, T; Cebrián-Pérez, J A

    2006-09-01

    This study was conducted to evaluate monthly changes in the ram seminal plasma protein profile using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) with a polyacrylamide linear gradient gel. Likewise, comparative analyses of the protein composition of ovine seminal plasma (SP) from ejaculates obtained along the year, and its relationship with sperm motility, viability and concentration of ejaculate were carried out. Western-blot analysis was performed to specifically detect P14, a ram SP protein postulated to be involved in sperm capacitation and gamete interaction [Barrios B, Fernández-Juan M, Muiño-Blanco T, Cebrián-Pérez JA. Immunocytochemical localization and biochemical characterization of two seminal plasma proteins which protect ram spermatozoa against cold-shock. J Androl 2005;26:539-49], and its variations along the year have also been established. The experiment was carried out from May 2003 to April 2004, with nine Rasa Aragonesa rams. Ejaculates obtained every 2 days were pooled and used for each assay, to avoid individual differences, and three two-dimensional SDS-PAGE gels were run for each month. The high resolution of the gradient gel allowed the image analysis software to detect around 252 protein spots, with pIs ranging from 4.2 to 7.6, and molecular weight (M(r)) from 12.5 to 83.9 kDa. Four protein spots (1, 2, 3 and 4) of low M(r) (15.1, 15.7, 15.9 and 21.0 kDa) and acidic pI (5.9, 5.3, 5.7 and 6.6), respectively, had the highest relative intensity in the SP map (11.2, 9.3, 4.7 and 7.7%, respectively). Spot 3 was more abundant (P<0.05) from May to December, and negatively correlated (P<0.05, r=-0.34) with sperm viability and concentration (P<0.05, r=0.36). Another 12 protein spots also had significant quantitative differences (P<0.05) along the year, and 17 protein spots, which correlated with some seminal quality parameter, did not show quantitative monthly changes. Western-blot analysis indicated that spots 1 and 2 reacted

  8. A high-definition native polyacrylamide gel electrophoresis system for the analysis of membrane complexes.

    PubMed

    Ladig, Roman; Sommer, Maik S; Hahn, Alexander; Leisegang, Matthias S; Papasotiriou, Dimitrios G; Ibrahim, Mohamed; Elkehal, Rajae; Karas, Michael; Zickermann, Volker; Gutensohn, Michael; Brandt, Ulrich; Klösgen, Ralf Bernd; Schleiff, Enrico

    2011-07-01

    Native polyacrylamide gel electrophoresis (PAGE) is an important technique for the analysis of membrane protein complexes. A major breakthrough was the development of blue native (BN-) and high resolution clear native (hrCN-) PAGE techniques. Although these techniques are very powerful, they could not be applied to all systems with the same resolution. We have developed an alternative protocol for the analysis of membrane protein complexes of plant chloroplasts and cyanobacteria, which we termed histidine- and deoxycholate-based native (HDN-) PAGE. We compared the capacity of HDN-, BN- and hrCN-PAGE to resolve the well-studied respiratory chain complexes in mitochondria of bovine heart muscle and Yarrowia lipolytica, as well as thylakoid localized complexes of Medicago sativa, Pisum sativum and Anabaena sp. PCC7120. Moreover, we determined the assembly/composition of the Anabaena sp. PCC7120 thylakoids and envelope membranes by HDN-PAGE. The analysis of isolated chloroplast envelope complexes by HDN-PAGE permitted us to resolve complexes such as the translocon of the outer envelope migrating at approximately 700 kDa or of the inner envelope of about 230 and 400 kDa with high resolution. By immunodecoration and mass spectrometry of these complexes we present new insights into the assembly/composition of these translocation machineries. The HDN-PAGE technique thus provides an important tool for future analyses of membrane complexes such as protein translocons.

  9. Mechanosensitivity of astrocytes on optimized polyacrylamide gels analyzed by quantitative morphometry

    NASA Astrophysics Data System (ADS)

    Moshayedi, Pouria; Costa, Luciano da F.; Christ, Andreas; Lacour, Stephanie P.; Fawcett, James; Guck, Jochen; Franze, Kristian

    2010-05-01

    Cells are able to detect and respond to mechanical cues from their environment. Previous studies have investigated this mechanosensitivity on various cell types, including neural cells such as astrocytes. In this study, we have carefully optimized polyacrylamide gels, commonly used as compliant growth substrates, considering their homogeneity in surface topography, mechanical properties, and coating density, and identified several potential pitfalls for the purpose of mechanosensitivity studies. The resulting astrocyte response to growth on substrates with shear storage moduli of G' = 100 Pa and G' = 10 kPa was then evaluated as a function of coating density of poly-D-lysine using quantitative morphometric analysis. Astrocytes cultured on stiff substrates showed significantly increased perimeter, area, diameter, elongation, number of extremities and overall complexity if compared to those cultured on compliant substrates. A statistically significant difference in the overall morphological score was confirmed with an artificial intelligence-based shape analysis. The dependence of the cells' morphology on PDL coating density seemed to be weak compared to the effect of the substrate stiffness and was slightly biphasic, with a maximum at 10-100 µg ml - 1 PDL concentration. Our finding suggests that the compliance of the surrounding tissue in vivo may influence astrocyte morphology and behavior.

  10. Sodium dodecyl sulfate-polyacrylamide gel protein electrophoresis of freshwater photosynthetic sulfur bacteria.

    PubMed

    Osuna, M Begoña; Casamayor, Emilio O

    2011-01-01

    Sodium dodecyl sulfate-polyacrylamide gel protein electrophoresis (SDS-PAGE) was carried out using different bacterial strains of the photosynthetic sulfur bacteria Chlorobium, Thiocapsa, Thiocystis, and Chromatium cultured in the laboratory, and the natural blooms in two karstic lakes (Lake Cisó and Lake Vilar, NE Spain) where planktonic photosynthetic bacteria (purple and green sulfur bacteria) massively developed accounting for most of the microbial biomass. Several extraction, solubilization, and electrophoresis methods were tested to develop an optimal protocol for the best resolution of the SDS-PAGE. Protein composition from different water depths and at different times of the year was visualized within a molecular mass range between 100 and 15 kDa yielding up to 20 different protein bands. Protein banding patterns were reproducible and changed in time and with depth in agreement with changes in photosynthetic bacteria composition. When a taxonomically stable community was followed in time, differences were observed in the intensity but not in the composition of the SDS-PAGE banding pattern. Three environmental variables directly related to the activity of sulfur bacteria (light, oxygen, and sulfide concentrations) had a significant effect on protein banding patterns and explained 33% of the variance. Changes in natural protein profiles of the bacterial blooms agreed with changes in species composition and in the in situ metabolic state of the populations.

  11. Fiber based optical tweezers for simultaneous in situ force exertion and measurements in a 3D polyacrylamide gel compartment.

    PubMed

    Ti, Chaoyang; Thomas, Gawain M; Ren, Yundong; Zhang, Rui; Wen, Qi; Liu, Yuxiang

    2015-07-01

    Optical tweezers play an important role in biological applications. However, it is difficult for traditional optical tweezers based on objective lenses to work in a three-dimensional (3D) solid far away from the substrate. In this work, we develop a fiber based optical trapping system, namely inclined dual fiber optical tweezers, that can simultaneously apply and measure forces both in water and in a 3D polyacrylamide gel matrix. In addition, we demonstrate in situ, non-invasive characterization of local mechanical properties of polyacrylamide gel by measurements on an embedded bead. The fiber optical tweezers measurements agree well with those of atomic force microscopy (AFM). The inclined dual fiber optical tweezers provide a promising and versatile tool for cell mechanics study in 3D environments.

  12. Stabilization of thin-layer agarose gels after isoelectric focusing with polyacrylamide enables reverse imidazole-zinc staining and facilitates two-dimensional gel electrophoresis.

    PubMed

    Hellman, Jukka

    2008-09-01

    Large-pore-size agarose gels provide excellent resolving capacity for high molecular weight biomolecules. Thin-layer agarose isoelectric focusing (IEF) gels on polyester support films are especially useful for the separation of large proteins based on their pI in native conformation, but the method has suffered from the lack of detection methods compatible with agarose gels in hydrated form. Recently, an acrylamide copolymerization method was reported to enable mass-spectrometry-compatible silver staining and in-gel digestion of proteins. In this study, the method was further applied by demonstrating successful reverse imidazole-zinc staining of thin-layer agarose IEF gels copolymerized with acrylamide. The sensitivity of the reverse staining method on the composite gel at its best equaled the sensitivity of the traditional dried agarose silver staining method. Owing to the increased durability and reversible detection, the reverse-stained first-dimension gel could be conveniently prepared for the second-dimension sodium dodecyl sulfate polyacrylamide gel electrophoresis by reduction and alkylation. In addition, the micropreparative generation of tryptic peptides of Coomassie brilliant blue R-250 stained proteins in the composite gel is demonstrated.

  13. Applicability of RAPD markers on silver-stained polyacrylamide gels to ascertain genetic diversity in Peripatus acacioi (Peripatidae; Onychophora).

    PubMed

    DeLaat, Daiane Mariele; Carvalho, Maria Raquel Santos; Lovato, Maria Bernadete; Acedo, Maria Dolores Porto; da Fonseca, Cleusa Graça

    2005-12-30

    RAPD (random amplification of polymorphic DNA) molecular markers can be utilized for analyzing genetic variability in populations for which only a few or no molecular markers are available. They were used in a study of an endangered species, Peripatus acacioi, found in the Tripuí Ecological Station, in Ouro Preto, MG, Brazil. The ecological station was specifically created to protect this velvet worm species, the first of this group found in Brazil. For an initial evaluation of the genetic diversity of this species, DNA samples from the lobopods of four individuals, collected at random, were analyzed using RAPD. Each reaction was run with a different primer (Operon RAPD 10-mer Kits), totaling 13 primers (OPC2, OPC3, OPC4, OPC6, OPC8, OPC10, OPC11, OPL2, OPL7, OPL11, OPL13, OPL18, and OPL19). Due to the low amplification yield, RAPD fragments were separated in polyacrylamide gels and stained with silver nitrate. Numerous bands were observed. Fifty-five of the amplified bands proved to be reproducible, both in terms of presence and intensity. Among these, 27 were variable and 28 were constant. The average number of bands per gel was 4.2. Nine of the 13 primers tested allowed the identification of constant and variable bands among these four individuals. RAPD analysis of genetic variation using silver-stained polyacrylamide gel electrophoresis provided measures of band sharing among the individuals, and therefore could be used in population genetics studies of P. acacioi.

  14. Nanopore density effect of polyacrylamide gel plug on electrokinetic ion enrichment in a micro-nanofluidic chip

    NASA Astrophysics Data System (ADS)

    Wang, Jun-yao; Xu, Zheng; Li, Yong-kui; Liu, Chong; Liu, Jun-shan; Chen, Li; Du, Li-qun; Wang, Li-ding

    2013-07-01

    In this paper, the nanopore density effect on ion enrichment is quantitatively described with the ratio between electrophoresis flux and electroosmotic flow flux based on the Poisson-Nernst-Planck equations. A polyacrylamide gel plug is integrated into a microchannel to form a micro-nanofluidic chip. With the chip, electrokinetic ion enrichment is relatively stable and enrichment ratio of fluorescein isothiocyanate can increase to 600-fold within 120 s at the electric voltage of 300 V. Both theoretical research and experiments show that enrichment ratio can be improved through increasing nanopore density. The result will be beneficial to the design of micro-nanofluidic chips.

  15. Separation and recovery of nucleic acids with improved biological activity by acid-degradable polyacrylamide gel electrophoresis.

    PubMed

    Kim, Yoon Kyung; Kwon, Young Jik

    2010-05-01

    One of the fundamental challenges in studying biomacromolecules (e.g. nucleic acids and proteins) and their complexes in a biological system is isolating them in their structurally and functionally intact forms. Electrophoresis offers convenient and efficient separation and analysis of biomacromolecules but recovery of separated biomacromolecules is a significant challenge. In this study, DNAs of various sizes were separated by electrophoresis in an acid-degradable polyacrylamide gel. Almost 100% of the nucleic acids were recovered after the identified gel bands were hydrolyzed under a mildly acidic condition and purified using anion exchange resin. Further concentration by centrifugal filtration and a second purification using ion exchange column chromatography yielded 44-84% of DNA. The second conventional (non-degradable) gel electrophoresis confirmed that the nucleic acids recovered from acid-degradable gel bands preserved their electrophoretic properties through acidic gel hydrolysis, purification, and concentration processes. The plasmid DNA recovered from acid-degradable gel transfected cells significantly more efficiently than the starting plasmid DNA (i.e. improved biological activity via acid-degradable PAGE). Separation of other types of nucleic acids such as small interfering RNA using this convenient and efficient technique was also demonstrated.

  16. Apolipoprotein distribution in human lipoproteins separated by polyacrylamide gradient gel electrophoresis.

    PubMed

    Vézina, C A; Milne, R W; Weech, P K; Marcel, Y L

    1988-05-01

    The heterogeneity of serum lipoproteins (excluding very low density (VLDL) and intermediate density (IDL) lipoproteins) and that of lipoproteins secreted by HepG2 cells has been studied by immunoblot analysis of the apolipoprotein composition of the particles separated by polyacrylamide gradient gel electrophoresis (GGE) under nondenaturing conditions. The reactions of antibodies to apoA-I, apoA-II, apoE, apoB, apoD, and apoA-IV have revealed discrete bands of particles which differ widely in size and apolipoprotein composition. GGE of native serum lipoproteins demonstrated that apoA-II is present in lipoproteins of limited size heterogeneity (apparent molecular mass 345,000 to 305,000) and that apoB is present in low density lipoproteins (LDL) and absent from all smaller or denser lipoproteins. In contrast, serum apoA-I, E, D, and A-IV are present in very heterogeneous particles. Serum apoA-I is present mainly in particles of 305 to 130 kDa where it is associated with apoA-II, and in decreasing order of immunoreactivity in particles of 130-90 kDa, 56 kDa, 815-345 kDa, and finally within the size range of LDL, all regions where there is little detectable apoA-II. Serum apoE is present in three defined fractions, one within the size range of LDL, one containing heterogeneous particles between 640 and 345 kDa, and one defined fraction at 96 kDa. Serum apoD is also present in three defined fractions, one comigrating with LDL, one containing heterogeneous particles between 390 and 150 kDa, and one band on the migration front. Most of serum apoA-IV is contained in a band comigrating with albumin. GGE of centrifugally prepared LDL shows the presence of apoB, apoE, and apoD, but not that of apoA-I. However, the particles containing apoA-I, which, in serum, migrated within the LDL size range and as bands of 815 to 345 kDa, were recovered upon centrifugation in the d greater than 1.21 g/ml fraction. GGE of high density lipoproteins (HDL) indicated that most of apoA-I, A

  17. Isolation and characterization of the pigment-protein complexes of Rhodopseudomonas sphaeroides by lithium dodecyl sulfate/polyacrylamide gel electrophoresis.

    PubMed

    Broglie, R M; Hunter, C N; Delepelaire, P; Niederman, R A; Chua, N H; Clayton, R K

    1980-01-01

    When purified photosynthetic membranes from Rhodopseudomonas sphaeroides were treated with lithium dodecyl sulfate and subjected to polyacrylamide gel electrophoresis at 4 degrees C, up to 11 pigment-protein complexes were resolved. Absorption spectra revealed that the smallest complex contained reaction center pigments and the others contained the antenna components B850 and B875 in various proportions. Of these antenna complexes, the largest was almost entirely B850 and the smallest contained only B875. After solubilization at 100 degrees C and electrophoresis on polyacrylamide gradient gels, the B850 complex gave rise to two polypeptide components migrating with apparent Mr of 10,000 and 8000, whereas with the B875 complex, two components were observed with apparent Mr of 12,000 and 8000. The reaction center complex gave rise to only the 24 and 21 kilodalton polypeptide subunits. Fluorescence emission spectra showed maxima at 872 and 902 nm for B850 and B875, respectively. Analyses of bacteriochlorophyll a and carotenoids indicated that, in the B875 complex, two molecules of each of these pigments are associated with the two polypeptides. The associations of B850 and B875 in large and small complexes obtained by lithium dodecyl sulfate treatment are consistent with models of their organization within the membrane.

  18. Mutation analysis of fragile X syndrome by Southern blot, radioactive PCR, silver-stained polyacrylamide gel and DIG DNA

    SciTech Connect

    Lee, Sook-Hwan; Kim, Un-Kyung; Chung-Woong, M.S.

    1994-09-01

    Fragile X syndrome is the most common inherited form of mental retardation. In fragile X syndrome, the underlying mutation is caused by an expansion of the CTG triplet in the 5{prime} untranslated region of the FMR-1 gene located at Xq27.3 and diagnosed by methylation of the associated CpG island. This disorder becomes clinically manifested when the mutation is caused by an expansion of (CGG)n reaching a threshold of about 600bp (200 repeats). The number of inserted repeats increases through the generation. We have analyzed fragile X syndrome by 4 different methods: Southern blot, radioactive PCR, polyacrylamide gel and DIG DNA labeling/detection techniques. Southern blot and DIG DNA labeling/detection by double DNA digestion with EcoRI and EagI reveals both the presence of the mutation and the methylation status. Radioactive PCR and silver-stained polyacrylamide gel is a rapid and sensitive technique to define the unaffected carriers and NTMs, but it is difficult to amplify such a highly GC-rich sequence. Further testing in other fragile X patients is currently in progress.

  19. A systematic investigation into the recovery of radioactively labeled proteins from sodium dodecyl sulfate-polyacrylamide gels.

    PubMed

    Zhou, Shaobo; Bailey, Matthew J; Dunn, Michael J; Preedy, Victor R; Emery, Peter W

    2004-01-01

    We report the results of a systematic investigation designed to optimize a method for quantifying radioactivity in proteins in sodium dodecyl sulfate-polyacrylamide gels. The method involves dissolving appropriately sized pieces of gel in hydrogen peroxide and heating to 70 degrees C overnight followed by liquid scintillation counting. H(2)O(2) had no effect on the count rates of [(14)C]bovine serum albumin (BSA) when counted in a conventional liquid scintillation system, and the count rates remained stable for several days. Temperatures below 70 degrees C resulted in incomplete extraction of radioactivity from gels containing [(14)C]BSA, but there was also a significant reduction in count rates in samples incubated at 80 degrees C. At 70 degrees C recovery was not affected by the amount of sample loaded onto the gel or by the staining procedure (Coomassie Brilliant Blue or SYPRO Ruby). Recoveries were in the range of 89-94%, and the coefficient of variation for five replicate samples was 5-10%. This method offers a reliable way of measuring the amount of radioactivity in proteins that have been separated by electrophoresis. It may be useful, for example, in quantitative metabolic labeling experiments when it is necessary to know precisely how much tracer has been incorporated into a particular protein.

  20. Resolution of 8-aminonaphthalene-1,3,6-trisulfonic acid-labeled glucose oligomers in polyacrylamide gel electrophoresis at low gel concentration.

    PubMed

    Cabanes-Macheteau, Marion; Chrambach, Andreas; Taverna, Myriam; Buzás, Zsuzsanna; Berna, Patrick

    2004-01-01

    A discontinuous Tris-Cl/acetate (OAc) buffer system, unprecedently containing OAc as the trailing constituent, and operative in polyacrylamide gel electrophoresis (PAGE) at low polyacrylamide concentration (T = 4.8%) is described in the paper. The characteristics of the electrophoretic system are illustrated by the resolution of fluorescent 8-aminonaphthalene-1,3,6-trisulfonic acid (ANTS)-labeled malto-oligosaccharides and dextran homopolymers. In this buffer system, the resolving phase is constituted by Tris-OAc behind a moving boundary formed between the leading chloride ion of Tris-HCl gel buffer and the trailing OAc ion provided by a catholyte of NH(4)OAc. In contrast with the results obtained with Tris-CI/glycinate buffer commonly used in electrophoresis, or with Tris-CI/borate, the best resolution of the glucose oligomers containing 1-4 glucose units in Tris-OAc, pH 8.8, ionic strength of 0.08, was obtained at 4.8% polyacrylamide concentration, using 0.5 M NH(4)OAc, pH 9.5 as the catholyte. Under those conditions, the ANTS-glucose oligomers were separated with mobilities decreasing from glucose to maltohexaose. The linear Ferguson plots (log relative mobility, R(f), vs.%T) of the glucose oligomers show that the surface net charge of those oligomers is inversely related to their sizes, given by the slopes, K(R), of the plots. The molecular weight of the oligomers is directly but nonlinearly related to K(R). The novel electrophoretic system illustrated here for separation of short ANTS-saccharides can be potentially applied to the resolution of other biomolecules such as rapidly migrating DNA, peptides or proteins.

  1. Isoelectric focusing of human hair keratins: correlation with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) patterns and effect of cosmetic treatments.

    PubMed

    Rodriguez-Calvo, M S; Carracedo, A; Muñoz, I; Concheiro, L

    1992-03-01

    A new isoelectric focusing (IEF) technique in polyacrylamide gels with 6M urea and 1.5% Nonidet P40 has been developed to characterize human hair samples. The phenotypes demonstrated with this procedure has been correlated with the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) patterns described by other authors. The method described can be applied in the forensic science analysis of a single human hair. Using the same IEF technique we have studied the changes in electrophoretic patterns of cosmetically treated hair. The characteristics of the modifications observed and its utility in forensic science work are also discussed in this paper.

  2. Silver stain for detecting 10-femtogram quantities of protein after polyacrylamide gel electrophoresis.

    PubMed

    Ohsawa, K; Ebata, N

    1983-12-01

    A rapid and highly sensitive silver stain and color stain were developed for visualizing proteins. The procedure is simple and the bands were clear. This silver stain detects 100 pg quantities of proteins. In order to stain quickly, sensitively, and sharply a protein matrix in a gel, the repeated shrinkage and swelling gel was developed with a hyper- and hypotonic solution to remove the sodium dodecyl sulfate (SDS) from SDS-protein complex and to generate influx of staining solution into the gel. We have found that the silver staining method with the repeated exposure to hyper- and hypotonic solution and a narrow well produced 10 fg order of proteins.

  3. Quantitation of yeast total proteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis sample buffer for uniform loading.

    PubMed

    Sheen, Hyukho

    2016-04-01

    Proteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) sample buffer are difficult to quantitate due to SDS and reducing agents being in the buffer. Although acetone precipitation has long been used to clean up proteins from detergents and salts, previous studies showed that protein recovery from acetone precipitation varies from 50 to 100% depending on the samples tested. Here, this article shows that acetone precipitates proteins highly efficiently from SDS-PAGE sample buffer and that quantitative recovery is achieved in 5 min at room temperature. Moreover, precipitated proteins are resolubilized with urea/guanidine, rather than with SDS. Thus, the resolubilized samples are readily quantifiable with Bradford reagent without using SDS-compatible assays.

  4. Rapid, simple method of preparing rotaviral double-stranded ribonucleic acid for analysis by polyacrylamide gel electrophoresis.

    PubMed Central

    Theil, K W; McCloskey, C M; Saif, L J; Redman, D R; Bohl, E H; Hancock, D D; Kohler, E M; Moorhead, P D

    1981-01-01

    A procedure for extracting rotaviral double-stranded ribonucleic acid (RNA) directly from fecal and intestinal specimens collected from calves and pigs is described. This procedure provides a rapid, simple, reproducible method of obtaining rotaviral double-stranded RNA preparations suitable for electrophoretic analysis in polyacrylamide-agarose composite gels. The rotaviral genome electrophoretic migration pattern produced by double-stranded RNA extracted directly from a specimen by this procedure was qualitatively identical to the electrophoretic migration pattern obtained with double-stranded RNA extracted from purified rotavirus derived from the same specimen. Direct extraction of specimens containing porcine rotavirus-like virus by this procedure gave preparations that had electrophoretic migration patterns similar, but not identical, to the characteristic electrophoretic migration pattern of the rotaviral genome. Sufficient rotaviral double-stranded RNA could be extracted from 6 ml of fecal or intestinal specimen by this procedure to permit 15 or more electrophoretic assays. Images PMID:6270190

  5. Rheological and mechanical behavior of polyacrylamide hydrogels chemically crosslinked with allyl agarose for two-dimensional gel electrophoresis.

    PubMed

    Suriano, R; Griffini, G; Chiari, M; Levi, M; Turri, S

    2014-02-01

    Two-dimensional (2-D) gel electrophoresis currently represents one of the most standard techniques for protein separation. In addition to the most commonly employed polyacrylamide crosslinked hydrogels, acrylamide-agarose copolymers have been proposed as promising systems for separation matrices in 2-D electrophoresis, because of the good resolution of both high and low molecular mass proteins made possible by careful control and optimization of the hydrogel pore structure. As a matter of fact, a thorough understanding of the nature of the hydrogel pore structure as well as of the parameters by which it is influenced is crucial for the design of hydrogel systems with optimal sieving properties. In this work, a series of acrylamide-based hydrogels covalently crosslinked with different concentrations of allyl agarose (0.2-1%) is prepared and characterized by creep-recovery measurements, dynamic rheology and tensile tests, in the attempt to gain a clearer understanding of structure-property relationships in crosslinked polyacrylamide-based hydrogels. The rheological and mechanical properties of crosslinked acrylamide-agarose hydrogels are found to be greatly affected by crosslinker concentration. Dynamic rheological tests show that hydrogels with a percentage of allyl agarose between 0.2% and 0.6% have a low density of elastically effective crosslinks, explaining the good separation of high molecular mass proteins in 2-D gel electrophoresis. Over the same range of crosslinker concentration, creep-recovery measurements reveal the presence of non-permanent crosslinks in the hydrogel network that justifies the good resolution of low molecular mass proteins as well. In tensile tests, the hydrogel crosslinked with 0.4% of allyl agarose exhibits the best results in terms of mechanical strength and toughness. Our results show how the control of the viscoelastic and the mechanical properties of these materials allow the design of mechanically stable hydrogels with improved

  6. Prestaining of glycoproteins in sodium dodecyl sulfate polyacrylamide gels by dansylhydrazine.

    PubMed

    Wang, Yang; Zhou, Xuan; Yu, Qing; Duan, Yuanmeng; Huang, Binbin; Hong, Guoying; Zhou, Ayi; Jin, Litai

    2014-06-01

    A new fluorescent prestaining method for gel-separated glycoproteins in 1D and 2D SDS-PAGE was developed by using dansylhydrazine in this study. The prestained gels could be easily imaged after electrophoresis without any time-consuming steps needed for poststains. As low as 4-8 ng glycoproteins (transferrin, α1-acid glycoprotein) could be selectively detected, which is comparable to that of Pro-Q Emerald 488, one of the most commonly used glycoprotein stain. In addition, a subsequent study of deglycosylation, glycoprotein affinity isolation, and LC-MS/MS analysis was performed to confirm the specificity of the newly developed method.

  7. Analysis of bacteriophage N protein and peptide binding to boxB RNA using polyacrylamide gel coelectrophoresis (PACE).

    PubMed Central

    Cilley, C D; Williamson, J R

    1997-01-01

    The antitermination protein N from bacteriophage lambda (Nlambda) interacts with the nut site in its own mRNA, as well as host factors, to facilitate formation of a termination-resistant transcription complex. The conserved, amino-terminal arginine-rich domain of Nlambda protein is known to interact with a small RNA hairpin (boxB) derived from the nut site RNA. We have examined the binding of Nlambda protein, peptides derived from the amino terminus of Nlambda, and the related phage P22 N protein to lambda boxB RNAs. To facilitate the study of complexes that are not amenable to gel retardation assays, a new polyacrylamide affinity coelectrophoresis technique (PACE) was developed. Using the PACE assay, we have demonstrated that a 19-amino acid peptide from the amino terminus of Nlambda protein binds lambda boxB RNA with a Kd,app of 5.2 nM. PACE was also used to study the binding affinity of a number of Nlambda peptide and lambda boxB RNA mutants. The PACE technique is complementary to the traditional gel retardation assay for direct measurement of binding interactions, and will be useful for any procedure that requires a pool of RNAs to be resolved based on their relative affinities for proteins or peptides. PMID:8990399

  8. Sensitive detection of proteins in polyacrylamide gel via isatoic anhydride derivatization: Introduction of a low-cost fluorescent prelabeling procedure.

    PubMed

    Asadollahi, Kazem; Rafiee, Saharnaz; Riazi, Gholamhossein

    2016-10-01

    Here, we introduce isatoic anhydride as a sensitive and commodious fluorescent prelabel for detection of proteins in one-dimensional polyacrylamide gels. High reactivity of isatoic anhydride with nucleophiles in mild alkaline environments makes it an appropriate tag for labeling of biomolecules. In this study, we show that preelectrophoresis labeling of proteins with isatoic anhydride for few minutes at room temperature allows detection of 2-4 ng of standard proteins, BSA and lysozyme, per band. Proteins were successfully labeled in the presence of a wide range of common biological reagents and in crude cell extract. The labeled proteins have the same electrophoretic migration in comparison to unlabeled proteins; however the application of saturation labeling method results in slight band broadening. Compatibility of the method with downstream processes was assessed by tryptic digestion of labeled proteins and study of peptide mixture using gel electrophoresis which revealed partial digestion of labeled proteins due to lysine modification. The present procedure is sensitive, rapid, and inexpensive and is a promising alternative for current protein staining procedures, where downstream processes are not desired.

  9. Use of Two-Dimensional Polyacrylamide Gel Electrophoresis to Identify and Classify Rhizobium Strains

    PubMed Central

    Roberts, Gary P.; Leps, Walter T.; Silver, Lin E.; Brill, Winston J.

    1980-01-01

    Fifty-seven strains of various Rhizobium species were analyzed by two-dimensional gel electrophoresis. Since the protein pattern on such gels is a reflection of the genetic background of the tested strains, similarities in pattern allowed us to estimate the relatedness between these strains. All group II rhizobia (slow growing) were closely related and were very distinct from group I rhizobia (fast growing). Rhizobium meliloti strains formed a distinct group. The collection of R. leguminosarum and R. trifolii strains together formed another distinct group. Although there were some similarities within the R. phaseoli, sesbania rhizobia, and lotus rhizobia, the members within these seemed much more diverse than the members of the above groups. The technique also is useful to determine whether two unknown strains are identical. Images PMID:16345514

  10. Detection of biotinylated proteins in polyacrylamide gels using an avidin-fluorescein conjugate.

    PubMed

    Nakamura, Michihiro; Tsumoto, Kouhei; Ishimura, Kazunori; Kumagai, Izumi

    2002-05-15

    Biotinylated proteins are widely used as a molecular tool in biotechnological applications. In this paper, we demonstrated that biotinylated proteins after electrophoresis were detected directly in gels using an avidin-fluorescein conjugate with a fluorescence image analyzer. Upon analysis of the purified and chemically biotinylated protein, the sensitivity of this method was almost equal to that of silver staining. Chemically biotinylated proteins of Escherichia coli cell surfaces could also be specifically detected with our method. Furthermore, recombinant proteins fused with the biotin acceptor domain and biotinylated enzymatically in vivo were also detected in a lysate of E. coli specifically. The sensitivity and specificity of our method are high, and the procedure is simple. Therefore, our method would benefit detection of biotinylated proteins via gel electrophoresis and also various fields of study using avidin-biotin technology.

  11. Improved staining of phosphoproteins with high sensitivity in polyacrylamide gels using Stains-All.

    PubMed

    Cong, Wei-Tao; Ye, Wei-Jian; Chen, Mao; Zhao, Ting; Zhu, Zhong-Xin; Niu, Chao; Ruan, Dan-Dan; Ni, Mao-Wei; Zhou, Xuan; Jin, Li-Tai

    2013-12-01

    An improved Stains-All (ISA) staining method for phosphoproteins in SDS-PAGE was described. Down to 0.5-1 ng phosphoproteins (α-casein, β-casein, or phosvitin) can be successfully selectively detected by ISA stain, which is approximately 120-fold higher than that of original Stains-All stain, but is similar to that of commonly used Pro-Q Diamond stain. Furthermore, unlike the original Stains-All protocol that was time consuming and light unstable, ISA stain could be completed within 60 min without resorting to protect the gels from light during the whole staining procedure. According to the results, it is concluded that ISA stain is a rapid, sensitive, specific, and economic staining method for a broad application to the research of phosphoproteins.

  12. An improved plant leaf protein extraction method for high resolution two-dimensional polyacrylamide gel electrophoresis and comparative proteomics.

    PubMed

    Alam, I; Sharmin, Sa; Kim, K-H; Kim, Y-G; Lee, Jj; Lee, B-H

    2013-02-01

    We report here a simple and universally applicable protocol for extracting high quality proteins from plant leaf tissues. The protocol provides improved resolution and reproducibility of two-dimensional polyacrylamide gel electrophoresis (2-DE) and reduces the time required to analyze samples. Partitioning rubisco by polyethylene glycol (PEG) fractionation provides clearer detection of low-abundance proteins. Co-extraction of interfering substances increases the sample conductivity, which results in poor electrophoretic separation. Re-extraction of PEG-fractionated samples with phenol effectively eliminated interfering substances, which results in optimal conductivity during separation in the first dimension of the isoelectric focusing. Smooth focusing reduces analysis time and provides superior resolution in 2-DE gels. Incubating the samples at -80° C instead of -20° C reduced protein precipitation time to 2-3 h. Removal of nonprotein contaminants and the use of sonication increased protein solubility without additional reagents. These changes enabled loading and separation of maximum amounts of proteins, which permitted improved protein identification by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). An immunological approach revealed that little or no ribulose-1, 5-bisphosphte bisphosphate carboxylase oxygenase was present in the PEG supernatant. In addition, low-abundance proteins, such as myelocytomatosis transcription factor (MYC) and alpha subunit of heterotrimeric guanine nucleotide-binding protein complex (Gα), were detected only in the modified PEG supernatant and not in the total protein. These results suggest that our protocol produced high quality proteins and made many low-abundant proteins available for proteomic analysis. The successful application of this protocol for analyzing the leaf proteomes of soybean, Miscanthus sinensis, barley, Chinese cabbage, peanut and tea (Camellia sinensis) suggests

  13. Genetic diversity analysis of faba bean (Vicia faba L.) germplasms using sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

    PubMed

    Hou, W W; Zhang, X J; Shi, J B; Liu, Y J

    2015-10-30

    To investigate genetic diversity and relationships of 101 faba bean (Vicia faba L.), landraces and varieties from different provinces of China and abroad were analyzed by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE). A total of 2625 unambiguous and stable bands from 101 germplasms were detected, and 36 different bands were classified according to the electrophoretic mobility patterns of the proteins as determined by the SDS-PAGE analysis, of which 16 were polymorphic. Besides the common bands, the protein bands of 92, 75, 62, 40, 34, 17, and 13 kDa presented the highest frequencies of 92.08, 90.10, 99.01, 95.05, 95.05, 98.02, and 95.05%, respectively. The other 29 polymorphic protein bands showed higher polymorphism with 16.09 polymorphic bands in average. The genetic similarity of the 101 genotypes tested varied from 0.6111 to 0.9722, with an average of 0.7122. Cluster analysis divided the 101 genotypes into six major clusters, which was consistent with the systematic classification of faba bean done in previous studies. The overall results indicated that SDS-PAGE was a useful tool for genetic diversity analysis and laid a solid foundation for future faba bean breeding.

  14. Analysis of cell wall extracts of Candida albicans by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot techniques.

    PubMed Central

    Ponton, J; Jones, J M

    1986-01-01

    Cell walls of intact yeast- and mycelial-phase Candida albicans B311 were extracted with different compounds: dithiothreitol, dithiothreitol with protease, dithiothreitol with lyticase, and dithiothreitol with protease followed by beta-glucuronidase with chitinase. Extracts were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot techniques. Dithiothreitol extracts contained the most satisfactory array of components for study. Analysis of these extracts demonstrated that the outer cell wall layers of Candida blastoconidia and germ tubes contained a complex array of polysaccharides, glycoproteins, and proteins. The proteins contributed to a latticework stabilized by covalent bonds that was important in determining the porosity of the outer cell wall layers. When equivalent weights were analyzed, mycelial-phase extract contained a more varied array of proteins than did yeast-phase extract. Only a portion of proteins in mycelial-phase extract elicited antibody responses in hyperimmunized rabbits or infected humans. A polysaccharide-rich, high-molecular-weight component (migrating at a position that would correspond to proteins having molecular weights of 235,000 to 250,000) and a protein component (molecular weight, 19,000) were readily demonstrable in the mycelial-phase extract but could not be identified in the yeast-phase extract. Images PMID:3527986

  15. Capillary zone electrophoresis of soil humic acid fractions obtained by coupling size-exclusion chromatography and polyacrylamide gel electrophoresis.

    PubMed

    Cavani, Luciano; Ciavatta, Claudio; Trubetskaya, Olga E; Reznikova, Olga I; Afanas'eva, Gaida V; Trubetskoj, Oleg A

    2003-01-03

    Capillary zone electrophoresis (CZE) was used for characterisation of soil humic acid (HA) fractions obtained by coupling size-exclusion chromatography with polyacrylamide gel electrophoresis, on the basis of their molecular size and electrophoretic mobility. CZE was conducted using several low alkaline buffers as background electrolyte (BGE): 50 mM carbonate, pH 9.0; 50 mM phosphate, pH 8.5; 50 mM borate, pH 8.3; 50 mM Tris-borate+1 mM EDTA+7 M urea+0.1% sodium dodecyl sulphate (SDS), pH 8.3. Independently of BGE conditions, the effective electrophoretic mobility of HA fractions were in good agreement with their molecular size. The better resolution of HA were obtained in Tris-borate-EDTA buffer with urea and SDS. This results indicated that CZE, mostly with BGE-contained disaggregating agents, is useful for separating HAs in fractions with different molecular sizes.

  16. Polyacrylamide gel disc electrophoresis of alkaline phosphatase isoenzymes in bone and liver disease.

    PubMed Central

    Warnes, T W; Hine, P; Kay, G

    1976-01-01

    Acrylamide gel disc electrophoresis provides a reliable and reasonably rapid method of differentiating the raised serum alkaline phosphatase (AP) of bone origin from that of liver origin. The technique has been placed for the first time on a semiquantitative basis. Measurement of both band width and band position effectively distinguishes the bone from the liver isoenzyme, but band width provides superior discrimination. An origin band was seen in none of the normal subjects and in only 7% of patients with bone disease but was present in 78% of patients with liver disease, a highly significant increase. Fifty percent of normal individuals had a small-intestinal band in serum taken two hours after a meal, as did 35% of patients with liver disease, but the incidence of intestinal bands in bone disease was only 11%, significantly less than in the other two groups. The genetic control of small-intestinal AP in serum has been confirmed, but it has been demonstrated that the decrease of intestinal AP in bone disorders is not genetically determined. Images PMID:977779

  17. Differences between fertilized and unfertilized chicken egg white proteins revealed by 2-dimensional gel electrophoresis-based proteomic analysis.

    PubMed

    Qiu, Ning; Liu, Wen; Ma, Meihu; Zhao, Lei; Li, Yuqi

    2013-03-01

    The egg white protein alterations during the early phase of chicken embryonic development were recently reported by our laboratory. Nevertheless, the original albumen differences between fresh unfertilized and fertilized chicken eggs have not been investigated. By using 2-dimensional gel electrophoresis (2-DE), coupled with matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF MS/MS) method, 1 ovalbumin protein spot as well as 6 ovalbumin-related protein Y spots were identified showing more than 10-fold differences (P < 0.01) in abundance between fresh unfertilized and fertilized chicken egg whites. Six of these protein spots represented higher intensity in fertilized eggs through 2-DE analysis. It was thus concluded that ovalbumin protein family, especially ovalbumin-related protein Y, may play an important role in embryonic development, which still needs to be validated. This finding will provide insight into embryogenesis to improve our understanding of the functions of ovalbumin family proteins in regulating or supporting embryonic development.

  18. Porcine salivary analysis by 2-dimensional gel electrophoresis in 3 models of acute stress: A pilot study

    PubMed Central

    Fuentes-Rubio, María; Cerón, José J.; de Torre, Carlos; Escribano, Damián; Gutiérrez, Ana M.; Tecles, Fernando

    2014-01-01

    The purpose of this research was to study changes in the salivary proteome of healthy pigs in stressful situations to identify any potential new salivary biomarker of stress. Three groups of animals were subjected to 3 stress models: snaring restraint followed by simulated sampling of vena cava blood; brief transport by road; and restriction of movement in a digestibility cage. Saliva was obtained from each animal before and 15 and 30 min after the induction of stress. The samples from the animals that showed the greatest increase in salivary cortisol concentration were pooled and run on 2-dimensional gels. Coomassie Brilliant Blue R-250 was used for spot detection and mass spectrometry for spot identification. Statistical analyses showed that 2 proteins had significant differences in expression before and after the induction of stress. These proteins were identified as odorant-binding protein and fragments of albumin. Further studies will be necessary to confirm the value of using these proteins as salivary biomarkers of stress in pigs. PMID:24688174

  19. Proteomic analysis of melanoma-derived exosomes by two-dimensional polyacrylamide gel electrophoresis and mass spectrometry.

    PubMed

    Mears, Rainy; Craven, Rachel A; Hanrahan, Sarah; Totty, Nick; Upton, Carol; Young, Sarah L; Patel, Poulam; Selby, Peter J; Banks, Rosamonde E

    2004-12-01

    Exosomes are 40-100 nm vesicles released by numerous cell types and are thought to have a variety of roles depending on their origin. Exosomes derived from antigen presenting cells have been shown to be capable of initiating immune responses in vivo and eradicating established tumours in murine models. Tumour-derived exosomes can be utilised as a source of tumour antigen for cross-priming to T-cells and are thus of interest for use in anti-tumour immunotherapy. Further exploration into the protein composition of exosomes may increase our understanding of their potential roles in vivo and this study has examined the proteome of exosomes purified from cell supernatants of the melanoma cell lines MeWo and SK-MEL-28. The vesicular nature and size (30-100 nm) of the purified exosomes was confirmed by electron microscopy and sucrose density gradient centrifugation. Western blotting demonstrated the absence of calnexin and cytochrome c, verifying the purity of the exosome preparations, as well as enrichment of MHC class I and the tumour-associated antigens Mart-1 and Mel-CAM. The two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) protein profiles of exosomes from the two cell lines were highly comparable and strikingly different from the profiles of the total cell lysates. Mass spectrometric sequencing identified proteins present in 49 protein spots in the exosome lysates. Several of these have been identified previously in exosomes but some are novel, including p120 catenin, radixin, and immunoglobulin superfamily member 8 (PGRL). Proteins present in whole-cell lysates that were significantly reduced or excluded from exosomes were also identified and included several mitochondrial and lysosomal proteins, again confirming the proposed endosomal origin of exosomes. This study presents a starting point for future more in-depth protein studies of tumour-derived exosomes which will aid the understanding of their biogenesis and targeting for use in anti

  20. Use of polyacrylamide gel moving boundary electrophoresis to enable low-power protein analysis in a compact microdevice.

    PubMed

    Duncombe, Todd A; Herr, Amy E

    2012-10-16

    In designing a protein electrophoresis platform composed of a single-inlet, single-outlet microchannel powered solely by voltage control (no pumps, values, injectors), we adapted the original protein electrophoresis format-moving boundary electrophoresis (MBE)-to a high-performance, compact microfluidic format. Key to the microfluidic adaptation is minimization of injection dispersion during sample injection. To reduce injection dispersion, we utilize a photopatterned free-solution-polyacrylamide gel (PAG) stacking interface at the head of the MBE microchannel. The nanoporous PAG molecular sieve physically induces a mobility shift that acts to enrich and sharpen protein fronts as proteins enter the microchannel. Various PAG configurations are characterized, with injection dispersion reduced by up to 85%. When employed for analysis of a model protein sample, microfluidic PAG MBE baseline-resolved species in 5 s and in a separation distance of less than 1 mm. PAG MBE thus demonstrates electrophoretic assays with minimal interfacing and sample handling, while maintaining separation performance. Owing to the short separation lengths needed in PAG MBE, we reduced the separation channel length to demonstrate an electrophoretic immunoassay powered with an off-the-shelf 9 V battery. The electrophoretic immunoassay consumed less than 3 μW of power and was completed in 30 s. To our knowledge, this is the lowest voltage and lowest power electrophoretic protein separation reported. Looking forward, we see the low-power PAG MBE as a basis for highly multiplexed protein separations (mobility shift screening assays) as well as for portable low-power diagnostic assays.

  1. SU-E-T-105: Development of 3D Dose Verification System for Volumetric Modulated Arc Therapy Using Improved Polyacrylamide-Based Gel Dosimeter

    SciTech Connect

    Ono, K; Fujimoto, S; Akagi, Y; Hirokawa, Y; Hayashi, S; Miyazawa, M

    2014-06-01

    Purpose: The aim of this dosimetric study was to develop 3D dose verification system for volumetric modulated arc therapy (VMAT) using polyacrylamide-based gel (PAGAT) dosimeter improved the sensitivity by magnesium chloride (MgCl{sub 2}). Methods: PAGAT gel containing MgCl{sub 2} as a sensitizer was prepared in this study. Methacrylic-acid-based gel (MAGAT) was also prepared to compare the dosimetric characteristics with PAGAT gel. The cylindrical glass vials (4 cm diameter, 12 cm length) filled with each polymer gel were irradiated with 6 MV photon beam using Novalis Tx linear accelerator (Varian/BrainLAB). The irradiated polymer gel dosimeters were scanned with Signa 1.5 T MRI system (GE), and dose calibration curves were obtained using T{sub 2} relaxation rate (R{sub 2} = 1/T{sub 2}). Dose rate (100-600 MU min{sup −1}) and fractionation (1-8 fractions) were varied. In addition, a cubic acrylic phantom (10 × 10 × 10 cm{sup 3}) filled with improved PAGAT gel inserted into the IMRT phantom (IBA) was irradiated with VMAT (RapidArc). C-shape structure was used for the VMAT planning by the Varian Eclipse treatment planning system (TPS). The dose comparison of TPS and measurements with the polymer gel dosimeter was accomplished by the gamma index analysis, overlaying the dose profiles for a set of data on selected planes using in-house developed software. Results: Dose rate and fractionation dependence of improved PAGAT gel were smaller than MAGAT gel. A high similarity was found by overlaying the dose profiles measured with improved PAGAT gel dosimeter and the TPS dose, and the mean pass rate of the gamma index analysis using 3%/3 mm criteria was achieved 90% on orthogonal planes for VMAT using improved PAGAT gel dosimeter. Conclusion: In-house developed 3D dose verification system using improved polyacrylamide-based gel dosimeter had a potential as an effective tool for VMAT QA.

  2. Isolation of a component from commercial coomassie brilliant blue R-250 that stains rubrophilin and other proteins red on polyacrylamide gels.

    PubMed

    Rosenthal, H L; Berger, R A; Tyler, A N; Moore, B W

    1988-05-12

    Commercially available Coomassie Brilliant Blue R-250 (C.I. 42660) is a popular and useful dye that stains most proteins blue on polyacrylamide gels. Some proteins from brain (rubrophilin), collagens, histones and parotid gland proteins are distinctly red when stained with Coomassie Blue. Commonly used Coomassie Brilliant Blue R-250 preparations may contain more than 30 distinct colored and fluorescent components that can be separated on silica gel chromatographic columns. A specific component has been isolated on silica gel columns that stains rubrophilin and other proline-rich proteins a reddish color. Fast atom bombardment mass spectrometry of the isolated rubrophilin staining principle indicates a molecular weight of 634 as compared to 826 for the major dye in the original Coomassie Brilliant Blue R-250. Infrared spectrometry is consistent with a difference between the rubrophilin staining principle and Coomassie Brilliant Blue R-250 of a toluene sulfonic acid residue.

  3. PhosphorImager enhancement of sedimentation equilibrium-quantitative polyacrylamide gel electrophoresis: a highly sensitive technique for quantitation of equilibrium gradients of individual components in mixtures.

    PubMed

    Darawshe, S; Merezhinskaya, N; Minton, A P

    1995-07-20

    The technique called sedimentation equilibrium-quantitative polyacrylamide gel electrophoresis (Darawshe et al. (1993) Anal. Biochem. 215, 236-242) has been extended to permit the quantitation and analysis of gradients of individual radiolabeled components in a mixture of radiolabeled solutes centrifuged to sedimentation equilibrium. Immediately following centrifugation, the contents of a sample tube are fractionated into aliquots corresponding to laminae of solution at different radial positions in the centrifuge. Following treatment with sodium dodecyl sulfate-containing buffer, a portion of each fraction is subjected to electrophoresis on a polyacrylamide gel. The gel is then incubated with a strong phosphor plate and subsequently scanned with a Molecular Dynamics PhosphorImage. The concentration of an individual radiolabeled component at a particular radial distance is proportional to the integrated intensity of the image of the radiolabeled band of that component in the fraction corresponding to that radial distance. Concentration gradients reconstructed in this fashion are interpreted in the context of conventional sedimentation equilibrium theory. The results of control experiments carried out with purified proteins of known molar mass and the measurement of the molar mass of a new, partially purified protein are reported.

  4. Analysis of Soluble Proteins in Natural Cordyceps sinensis from Different Producing Areas by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis and Two-dimensional Electrophoresis

    PubMed Central

    Li, Chun-Hong; Zuo, Hua-Li; Zhang, Qian; Wang, Feng-Qin; Hu, Yuan-Jia; Qian, Zheng-Ming; Li, Wen-Jia; Xia, Zhi-Ning; Yang, Feng-Qing

    2017-01-01

    Background: As one of the bioactive components in Cordyceps sinensis (CS), proteins were rarely used as index components to study the correlation between the protein components and producing areas of natural CS. Objective: Protein components of 26 natural CS samples produced in Qinghai, Tibet, and Sichuan provinces were analyzed and compared to investigate the relationship among 26 different producing areas. Materials and Methods: Proteins from 26 different producing areas were extracted by Tris-HCl buffer with Triton X-100, and separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional electrophoresis (2-DE). Results: The SDS-PAGE results indicated that the number of protein bands and optical density curves of proteins in 26 CS samples was a bit different. However, the 2-DE results showed that the numbers and abundance of protein spots in protein profiles of 26 samples were obviously different and showed certain association with producing areas. Conclusions: Based on the expression values of matched protein spots, 26 batches of CS samples can be divided into two main categories (Tibet and Qinghai) by hierarchical cluster analysis. SUMMARY The number of protein bands and optical density curves of proteins in 26 Cordyceps sinensis samples were a bit different on the sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profilesNumbers and abundance of protein spots in protein profiles of 26 samples were obvious different on two-dimensional electrophoresis mapsTwenty-six different producing areas of natural Cordyceps sinensis samples were divided into two main categories (Tibet and Qinghai) by Hierarchical cluster analysis based on the values of matched protein spots. Abbreviations Used: SDS-PAGE: Sodium dodecyl sulfate polyacrylamide gel electrophoresis, 2-DE: Two-dimensional electrophoresis, Cordyceps sinensis: CS, TCMs: Traditional Chinese medicines PMID:28250651

  5. Simultaneous immunoblotting analysis with activity gel electrophoresis and 2-D gel electrophoresis.

    PubMed

    Lee, Der-Yen; Chang, Geen-Dong

    2015-01-01

    Diffusion blotting method can couple immunoblotting analysis with another biochemical technique in a single polyacrylamide gel, however, with lower transfer efficiency as compared to the conventional electroblotting method. Thus, with diffusion blotting, protein blots can be obtained from an SDS polyacrylamide gel for zymography assay, from a native polyacrylamide gel for electrophoretic mobility shift assay (EMSA) or from a 2-D polyacrylamide gel for large-scale screening and identification of a protein marker. Thereafter, a particular signal in zymography, electrophoretic mobility shift assay, and 2-dimensional gel can be confirmed or identified by simultaneous immunoblotting analysis with a corresponding antiserum. These advantages make diffusion blotting desirable when partial loss of transfer efficiency can be tolerated or be compensated by a more sensitive immunodetection reaction using enhanced chemiluminescence detection.

  6. Nondenaturing polyacrylamide gel electrophoresis to study the dissociation of the p53·MDM2/X complex by potentially anticancer compounds.

    PubMed

    Sgammato, Roberta; Desiderio, Doriana; Lamberti, Anna; Raimo, Gennaro; Novellino, Ettore; Carotenuto, Alfonso; Masullo, Mariorosario

    2015-12-01

    A new analytical method to study the dissociation of the complexes between the oncosuppressor p53 and its negative modulators murine double-minute protein 2 (MDM2) or MDMX, is proposed. This technique is reliable to determine the dissociative power exerted by small molecules on the complex taking advantage of the appearance of migrating MDM2 or MDMX in a native polyacrylamide gel, when inhibitors are added to the complex mixture. Therefore, we propose this new approach to easily screen library of compounds, with potential pharmacological anticancer activity.

  7. Fluid diversion and sweep improvement with chemical gels in oil recovery processes. [Four types of gels: resorcinol-formaldehyde; colloidal silica; Cr sup 3+ (chloride)-xanthan; and Cr sup 3+ (acetate)-polyacrylamide

    SciTech Connect

    Seright, R.S.; Martin, F.D.

    1992-09-01

    The objectives of this project were to identify the mechanisms by which gel treatments divert fluids in reservoirs and to establish where and how gel treatments are best applied. Several different types of gelants were examined, including polymer-based gelants, a monomer-based gelant, and a colloidal-silica gelant. This research was directed at gel applications in water injection wells, in production wells, and in high-pressure gas floods. The work examined how the flow properties of gels and gelling agents are influenced by permeability, lithology, and wettability. Other goals included determining the proper placement of gelants, the stability of in-place gels, and the types of gels required for the various oil recovery processes and for different scales of reservoir heterogeneity. During this three-year project, a number of theoretical analyses were performed to determine where gel treatments are expected to work best and where they are not expected to be effective. The most important, predictions from these analyses are presented. Undoubtedly, some of these predictions will be controversial. However, they do provide a starting point in establishing guidelines for the selection of field candidates for gel treatments. A logical next step is to seek field data that either confirm or contradict these predictions. The experimental work focused on four types of gels: (1) resorcinol-formaldehyde, (2) colloidal silica, (3) Cr{sup 3+}(chloride)-xanthan, and (4) Cr{sup 3+}(acetate)-polyacrylamide. All experiments were performed at 41{degrees}C.

  8. Analysis of HeLa cell hypoxanthine phosphoribosyltransferase mutants and revertants by two-dimensional polyacrylamide gel electrophoresis: evidence for silent gene activation.

    PubMed Central

    Milman, G; Lee, E; Ghangas, G S; McLaughlin, J R; George, M

    1976-01-01

    The spot corresponding to hypoxanthine phosphoribosyltransferase (HPRT; IMP:pyrophosphate phosphoribosyltransferase, EC 2.4.2.8) has been identified in two-dimensional polyacrylamide gels of HeLa cell extracts. This spot is absent in gels of 24 HPRT dificient mutants. A missense mutant displays a new HPRT spot at the same molecular weight but different isoelectric focusing position. Five independently isolated revertants of the missense mutant display spots corresponding to both the wild-type and mutant proteins indicating that they synthesize HPRT from two separate genes. If the missense protein is synthesized from a mutated form of the initially active HPRT gene, then wild-type HPRT protein in the revertants must be snythesized from a newly activated but prevously silent wild-type gene. The newly activated gene in the revertants of the missense mutation appears unstable producing a high frequency of spontaneous HPRT mutants. Images PMID:63948

  9. Protein/RNA coextraction and small two-dimensional polyacrylamide gel electrophoresis for proteomic/gene expression analysis of renal cancer biopsies.

    PubMed

    Barbero, Giovanna; Carta, Franco; Giribaldi, Giuliana; Mandili, Giorgia; Crobu, Salvatore; Ceruti, Carlo; Fontana, Dario; Destefanis, Paolo; Turrini, Francesco

    2006-02-01

    A small amount of bioptic tissue ( approximately 5-10mg of fresh tissue) usually does not contain enough material to extract protein and RNA separately, to obtain preparative two-dimensional polyacrylamide gel electrophoresis (2-DE), and to identify a large number of separated proteins by MS. We tested a method, on small renal cancer specimens, for the coextraction of protein and RNA coupled with 2-DE and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) or quadrupole time-of-flight (Q-TOF) analysis. We coextracted 0.28+/-0.05mg of proteins and 2.5+/-0.33microg of RNA for each 10mg of renal carcinoma tissue. Small and large 2-DE gels were compared: they showed a similar number of spots, and it was possible to match each other; using small format gels, one-fifth of the protein amount was required to identify, by Q-TOF analysis, the same number of proteins identifiable in large-format gel using MALDI-TOF analysis. Quality of RNA coextracted with the proteins was tested by real-time PCR on a set of housekeeping genes. They were quantified with high amplification efficiency and specificity. In conclusion, using 5 to 10mg of fresh tissue, it was possible to perform comprehensive parallel proteomic and genomic analysis by high-resolution, small-format 2-DE gels, allowing approximately 300 proteins identification and 1000 genes expression analysis.

  10. Phosphohydrolase activity of the isolated, brush-border membrane of Hymenolepis diminuta (Cestoda) following sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis.

    PubMed

    Pappas, P W

    1980-12-01

    Following electrophoresis of isolated, brush-border membranes of Hymenolepis diminuta on SDS-polyacrylamide gels, three distinct areas of alpha-naphthyl phosphate (NP) hydrolysis were detected; these corresponded to proteins with molecular weights of 106,800, 172,700, and greater than 340,000 Daltons. Hydrolysis of NP was inhibited by adenosine triphosphate, adenosine;5'-monophosphate, p-nitrophenyl-phosphate, glucose-1-phosphate, glucose-6-phosphate, fructose-6-phosphate, fructose-1,6-diphosphate, molybdate, ethylenediaminetetraacetate (EDTA), and ethyleneglycol-bis-(beta-amino-ethyl)-N,N'-tetraacetate (EGTA), but not by fluoride. Inhibition of NP hydrolysis by EDTA was relieved in the presence of Mg++ or Ca++. Heating the isolated, brush-border membrane in the presence of SDS for 5 min at 95 C destroyed all enzymatic activity. These characteristics indicated that the enzyme(s) responsible for NP hydrolysis (following separation of membrane proteins by SDS-polyacrylamide gel electrophoresis) were the same enzymes responsible for the phosphohydrolase activity associated with intact and solubilized, brush-border membrane preparations of H. diminuta.

  11. Identification and comparative proteomic study of quail and duck egg white protein using 2-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry analysis.

    PubMed

    Hu, S; Qiu, N; Liu, Y; Zhao, H; Gao, D; Song, R; Ma, M

    2016-05-01

    A proteomic study of egg white proteins from 2 major poultry species, namely quail (Coturnix coturnix) and duck (Anas platyrhynchos), was performed with comparison to those of chicken (Gallus gallus) through 2-dimensional polyacrylamide gel electrophoresis (2-DE) analysis. By using matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF MS/MS), 29 protein spots representing 10 different kinds of proteins as well as 17 protein spots designating 9 proteins were successfully identified in quail and duck egg white, respectively. This report suggested a closer relationship between quail and chicken egg white proteome patterns, whereas the duck egg white protein distribution on the 2-DE map was more distinct. In duck egg white, some well-known major proteins, such as ovomucoid, clusterin, extracellular fatty acid-binding protein precursor (ex-FABP), and prostaglandin D2 synthase (PG D2 synthase), were not detected, while two major protein spots identified as "deleted in malignant brain tumors 1" protein (DMBT1) and vitellogenin-2 were found specific to duck in the corresponding range on the 2-DE gel map. These interspecies diversities may be associated with the egg white protein functions in cell defense or regulating/supporting the embryonic development to adapt to the inhabiting environment or reproduction demand during long-term evolution. The findings of this work will give insight into the advantages involved in the application on egg white proteins from various egg sources, which may present novel beneficial properties in the food industry or related to human health.

  12. Fluorographic detection of tritiated glycopeptides and oligosaccharides separated on polyacrylamide gels: analysis of glycans from Dictyostelium discoideum glycoproteins

    SciTech Connect

    Prem Das, O.; Henderson, E.J.

    1986-11-01

    Previous workers have shown that oligosaccharides and glycopeptides can be separated by electrophoresis in buffers containing borate ions. However, normal fluorography of tritium-labeled structures cannot be performed because the glycans are soluble and can diffuse during equilibration with scintillants. This problem has been circumvented by equilibration of the gel with 2,5-diphenyloxazole (PPO) prior to electrophoresis. The presence of PPO in the gel during electrophoresis does not alter mobility of the glycopeptides and oligosaccharides. After electrophoresis, the gel is simply dried and fluorography performed. This allows sensitive and precise comparisons of labeled samples in parallel lanes of a slab gel and, since mobilities are highly reproducible, between different gels. The procedure is preparative in that after fluorography the gel bands can be quantitatively eluted for further study, without any apparent modification by the procedure. In this report, the procedure is illustrated by fractionation of both neutral and anionic glycopeptides produced by the cellular slime mold Dictyostelium discoideum.

  13. A facile approach to fabricate porous nanocomposite gels based on partially hydrolyzed polyacrylamide and cellulose nanocrystals for adsorbing methylene blue at low concentrations.

    PubMed

    Zhou, Chengjun; Lee, Sunyoung; Dooley, Kerry; Wu, Qinglin

    2013-12-15

    Porous nanocomposite gels were fabricated by a facile method consisting of the electrospinning and subsequent heat treatment based on partially hydrolyzed polyacrylamide (HPAM) of ultra-high molecular weight, with cellulose nanocrystals (CNCs) as crosslinker. The effects of three electrospinning parameters (i.e., solution concentration, composition of solvent mixture, and CNC loading level) on morphology and diameter of electrospun fibers were systematically investigated. The swelling properties of porous gels and their application in the removal of methylene blue dye (as a compound representative of contaminants) were evaluated. Electrospun fiber morphologies without beads, branches, and ribbons were achieved by optimizing the electrospinning solutions. The thermal crosslinking between HPAM and CNCs was realized through esterification, rendering the product nanocomposite membranes insoluble in water. Electrospun fibers of approximately 220 nm in diameter comprised the 3D porous nanocomposite gels, with porosity greater than 50%. The porous nanocomposite gels displayed a rapid swelling rate and an efficient adsorption capacity in removing methylene blue at low concentrations from aqueous solutions.

  14. Coupling sodium dodecyl sulfate-capillary polyacrylamide gel electrophoresis with matrix-assisted laser desorption ionization time-of-flight mass spectrometry via a poly(tetrafluoroethylene) membrane.

    PubMed

    Lu, Joann J; Zhu, Zaifang; Wang, Wei; Liu, Shaorong

    2011-03-01

    Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) is a fundamental analytical technique for proteomic research, and SDS-capillary gel electrophoresis (CGE) is its miniaturized version. Compared to conventional slab-gel electrophoresis, SDS-CGE has many advantages such as increased separation efficiency, reduced separation time, and automated operation. SDS-CGE is not widely accepted in proteomic research primarily due to the difficulties in identifying the well-resolved proteins. MALDI-TOF-MS is an outstanding platform for protein identifications. Coupling the two would solve the problem but is extremely challenging because the MS detector has no access to the SDS-CGE-resolved proteins and the SDS interferes with MS detection. In this work we introduce an approach to address these issues. We discover that poly(tetrafluoroethylene) (PTFE) membranes are excellent materials for collecting SDS-CGE-separated proteins. We demonstrate that we can wash off the SDS bound to the collected proteins and identify these proteins on-membrane with MALDI-TOF-MS. We also show that we can immunoblot and Coomassie-stain the proteins collected on these membranes.

  15. Use of fluorescein hydrazide and fluorescein thiosemicarbazide reagents for the fluorometric determination of protein carbonyl groups and for the detection of oxidized protein on polyacrylamide gels.

    PubMed

    Ahn, B; Rhee, S G; Stadtman, E R

    1987-03-01

    Highly fluorescent thiosemicarbazide and hydrazide prepared by reaction of fluorescein isothiocyanate with hydrazine or adipic acid dihydrazide have been used to monitor the presence of carbonyl groups in oxidatively modified proteins. After oxidation, proteins react with these reagents under anaerobic conditions in the dark to yield fluorescent protein conjugates (presumably thiosemicarbazones or hydrazones) which can be visualized as fluorescent bands following electrophoresis (0-4 degrees C) on lithium dodecyl sulfate-polyacrylamide gels. These reagents do not react with unoxidized proteins. The conjugates formed dissociate readily at room temperature but are fairly stable at pH 6-9, 0 degrees C. Current data suggest that these reagents will be useful in the detection and quantitation of oxidatively modified proteins in biological systems.

  16. Determination of the molecular weight of human gamma-3 chains by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate

    PubMed Central

    Virella, G.; Parkhouse, R. M. E.

    1972-01-01

    The molecular weights (mol. wt) for heavy chains of human IgG were estimated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate. Polyclonal IgG and monoclonal IgG proteins of different subclasses were extensively reduced with 50 mM dithioerythritol, in the presence of 2 per cent sodium dodecyl sulphate, at 100°. Four control proteins of known mol. wt (cytochrome C, chymotrypsinogen A, egg albumin, and serum albumin) were used to construct a linear plot of electrophoretic mobility versus log mol. wt. From this plot, the following mol. wts were calculated: 53,650±700 for polyclonal IgG; 54,200±1065 for γ1, γ2, and γ4 chains, and 60,950±585 for γ3 chains. Those results confirm the larger size of γ3 chains reported by Saluk and Clem (1971). PMID:4346255

  17. Analysis of polyacrylamide gels for trace metals using diffusive gradients in thin films and laser ablation inductively coupled plasma mass spectrometry.

    PubMed

    Warnken, Kent W; Zhang, Hao; Davison, William

    2004-10-15

    A simple method for the analysis of polyacrylamide diffusive gradients in thin film (DGT) gels by laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS), employing a novel use of (115)In internal standardization, has been developed. This method allows the determination of Co, Ni, Cu, Zn, Cd, and Pb concentrations (at the DGT filter face) or fluxes in sediments at a spatial resolution of 100 microm. Single-layered gels, using an optimized laser defocus of 4000 microm at 400 mJ power, showed high precision (generally approximately 10%) and a linear response during solution deployment. Of the elements Sc, In, Ba, La, Ce, and Tb, Ba most closely tracked variations in laser energy and showed the highest analytical precision but could not be used as an internal standard due to its elevated presence in natural sediments. Therefore, internal standardization, necessary to normalize data collected on different days, was carried out using (115)In contained within a second layer of backing gel and dried along with the analyte layer as a dual-gel disk. This multilayered gel standard required a laser defocus setting of 1000 microm and a laser power of approximately 800 mJ. Analytical precision for a 64-spot ablation grid at 100-microm spacing was approximately 10%. Verification of this method was carried out on DGT sediment probes deployed in Priest Pot (English Lake District). Results obtained by conventional slicing techniques and aqueous elution agreed with laser ablation results when the different sampling areas were considered. The elution results varied by a factor of <2, whereas the laser ablation technique showed a variability of approximately 4, indicating localized elevated concentrations of Co. This higher resolution LA-ICPMS method could ultimately lead to an improved understanding of the geochemical processes responsible for metal uptake and release in sediments.

  18. Detection of metals in proteins by means of polyacrylamide gel electrophoresis and laser ablation-inductively coupled plasma-mass spectrometry: application to selenium.

    PubMed

    Chéry, Cyrille C; Günther, Detlef; Cornelis, Rita; Vanhaecke, Frank; Moens, Luc

    2003-10-01

    The capabilities of laser ablation-inductively coupled plasma-mass spectrometry for the detection of trace elements in a gel after gel electrophoresis were systematically studied. Figures of merit, such as limit of detection, linearity, and repeatability, were evaluated for various elements (Li, V, Cr, Mn, Ni, Cu, Zn, As, Se, Mo, Pd, Ag, Cd, Pt, Tl, Pb). Two ablation strategies were followed: single hole drilling, relevant for ablation of spots after two-dimensional (2-D) separations, and ablation with translation, i.e., on a line, relevant for one-dimensional (1-D) separations. This technique was applied to the detection of selenoproteins in red blood cells extracts after a 1-D separation (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and the detection of selenium-containing proteins in yeast after 2-D electrophoresis (2-DE). The detection procedure was further improved by using the dynamic reaction cell technology, which allowed the removal of the Ar_2(+) interference and hence the use of the most abundant Se isotope, (80)Se. Reaction gases were compared (methane, carbon monoxide, ammonia, oxygen and the combination of argon (collision gas) and hydrogen (reaction gas)). In each instance, the reaction cell parameters were optimized in order to obtain the lowest detection limit for Se (as (80)Se(+), (82)Se(+) or (77)Se(+); and as (80)Se(16)O(+), (82)Se(16)O(+) or (77)Se(16)O(+) with O(2) as the reaction gas). Carbon monoxide was found to offer the best performance. The detection limit with the use of DRC and He as transport gas was 0.07 microg Se g(-1) gel with single hole drilling and 0.15 microg Se g(-1) gel for ablation with translation.

  19. An optical comparator for measuring two-dimensional polyacrylamide gel electrophoresis records using an on-line microcomputer.

    PubMed

    Spragg, S P; Jones, M I; Hill, B J

    1983-03-01

    A comparator which makes it possible to compare two wet gels or photographic negatives or autoradiograms through a flickering light system has been built. The system consists of two special-purpose projectors which combine the images on a digitizing platform. When the lights are switched On and off out of phase, the positions of the common components remain unchanged, whereas those that are spatially displaced appear to jump from side to side and those present in one image but not the other switch on and off. This produces a flickering image in which differences are readily seen. Commercial camera lenses were used to construct the projectors and the overall specifications for the system are given. The coordinates of both the displaced components, as well as the selected standards from the two images, are digitized and entered automatically into an on-line microcomputer. By using an iterative procedure for collecting records from several superimposable records of the gel, it is possible to compensate for the lack of total reproducibility over the whole gels. These coordinates are then normalized and superimposed on a master map through a television display using a curser to adjust the coordinates. The whole procedure can be repeated for many gels using a common reference gel in the comparator, and the result is a set of normalized coordinates which can be plotted on a single map to provide a final record of the experiments.

  20. Comparison of diazo-coupling, formazan, and silver staining techniques for visualizing alkaline phosphatase isoenzymes after electrophoresis in homogeneous-pore and gradient-pore polyacrylamide gels.

    PubMed

    Hodson, A W; Skillen, A W

    1988-03-01

    Three techniques for visualization of alkaline phosphatase after polyacrylamide-gel electrophoresis are compared. These are diazo-dye simultaneous coupling with the substrate sodium naphthyl phosphate and 5-chloro-2-toluene diazonium chloride; formazan precipitation with the substrate 5-bromo-4-chloro-3-indolyl phosphate and 3-[4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide; and silver staining with the substrate sodium glycerophosphate. Each staining technique was tested with gradient-pore and homogeneous-pore acrylamide-gel electrophoresis. The main factors assessed are sensitivity; separation of the human serum alkaline phosphatase isoenzymes of the liver, bone, and intestinal types; and differences in substrate affinity, as well as the complexity of each technique. Using the three techniques only minor differences in substrate affinity are evident. There is some nonspecific staining with the diazo-coupling technique but not with the formazan and silver staining techniques. The differences, in the mobility of the liver, bone, and intestinal isoenzymes achieved by homogeneous-pore gel electrophoresis are sufficient to allow them to be clearly distinguished. However, only very small differences in mobility are found with gradient-pore gel electrophoresis, but the sharper bands in this medium allow much smaller amounts of activity to be detected. As little as 160 microU of enzyme can be visualized by the diazo technique. Silver staining gives an approximately fourfold increase in sensitivity over the formazan technique, which in turn gives a fourfold increase over the diazo technique. An important aspect of the silver staining technique is the potential of increasing sensitivity much further by improvements in the photographic physical development stage.(ABSTRACT TRUNCATED AT 250 WORDS)

  1. System and method of infrared matrix-assisted laser desorption/ionization mass spectrometry in polyacrylamide gels

    DOEpatents

    Haglund, Jr., Richard F.; Ermer, David R.; Baltz-Knorr, Michelle Lee

    2004-11-30

    A system and method for desorption and ionization of analytes in an ablation medium. In one embodiment, the method includes the steps of preparing a sample having analytes in a medium including at least one component, freezing the sample at a sufficiently low temperature so that at least part of the sample has a phase transition, and irradiating the frozen sample with short-pulse radiation to cause medium ablation and desorption and ionization of the analytes. The method further includes the steps of selecting a resonant vibrational mode of at least one component of the medium and selecting an energy source tuned to emit radiation substantially at the wavelength of the selected resonant vibrational mode. The medium is an electrophoresis medium having polyacrylamide. In one embodiment, the energy source is a laser, where the laser can be a free electron laser tunable to generate short-pulse radiation. Alternatively, the laser can be a solid state laser tunable to generate short-pulse radiation. The laser can emit light at various ranges of wavelength.

  2. Polyacrylamide gel substrates that simulate the mechanical stiffness of normal and malignant neuronal tissues increase protoporphyin IX synthesis in glioma cells

    NASA Astrophysics Data System (ADS)

    Niu, Carolyn J.; Fisher, Carl; Scheffler, Kira; Wan, Rachel; Maleki, Hoda; Liu, Haijiao; Sun, Yu; Simmons, Craig A.; Birngruber, Reginald; Lilge, Lothar

    2015-09-01

    Protoporphyrin IX (PPIX) produced following the administration of exogenous 5d-aminolevulinic acid is clinically approved for photodynamic therapy and fluorescence-guided resection in various jurisdictions around the world. For both applications, quantification of PPIX forms the basis for accurate therapeutic dose calculation and identification of malignant tissues for resection. While it is well established that the PPIX synthesis and accumulation rates are subject to the cell's biochemical microenvironment, the effect of the physical microenvironment, such as matrix stiffness, has received little attention to date. Here we studied the proliferation rate and PPIX synthesis and accumulation in two glioma cell lines U373 and U118 cultured under five different substrate conditions, including the conventional tissue culture plastic and polyacrylamide gels that simulated tissue stiffness of normal brain (1 kPa) and glioblastoma tumors (12 kPa). We found that the proliferation rate increased with substrate stiffness for both cell lines, but not in a linear fashion. PPIX concentration was significantly higher in cells cultured on tissue-simulating gels than on the much stiffer tissue culture plastic for both cell lines. These findings, albeit preliminary, suggest that the physical microenvironment might be an important determinant of tumor aggressiveness and PPIX synthesis in glioma cells.

  3. Light Scattering Induced Giant Red-Shift in Photoluminescence from CdTe Quantum Dots Encapsulated in Polyacrylamide Gel Nanospheres

    NASA Astrophysics Data System (ADS)

    Garner, Brett W.; Cai, Tong; Hu, Zhibing; Kim, Moon; Neogi, Arup

    2009-07-01

    The photoluminescence emission from CdTe quantum dots embedded in hydrogel nanospheres based on poly(N-isopropylacrylamide) (PNIPAM) polymer is observed to be modified by the random light scattering within the colloidal medium. Photoluminescence emission from CdTe quantum dots of various size has been observed making the gel fluorescent. The optical properties of the quantum dots entrapped within the gel microspheres can be modified due to change in refractive index, volume density of the surrounding hydrogel medium. A red-shift of ˜100 nm has been observed from quantum dots emitting in the green wavelength region as the cell length is increased. This shift is due to secondary scattering and energy transfer induced by the larger scattering cross-section within the medium which results in a re-excitation of larger sized quantum dots.

  4. A rapid and simple 8-quinolinol-based fluorescent stain of phosphoproteins in polyacrylamide gel after electrophoresis.

    PubMed

    Wang, Xu; Hwang, Sun-Young; Cong, Wei-Tao; Jin, Li-Tai; Choi, Jung-Kap

    2015-10-01

    In order to obtain an easy and rapid protocol to visualize phosphoproteins in SDS-PAGE, a fluorescent detection method named 8-Quinolinol (8-Q) stain is described. 8-Q can form ternary complexes in the gel matrix contributed by the affinity of aluminum ion (Al(3+) ) to the phosphate groups on the proteins and the metal chelating property of 8-Quinolinol, exhibiting strong fluorescence in ultraviolet light. It can visualize as little as 4∼8 ng of α-casein and β-casein, 16∼32 ng of ovalbumin and κ-casein which is more sensitive than Stains-All but less sensitive than Pro-Q Diamond. The protocol of 8-Q requires only 70 min in 0.75 mm mini-size or 1.0 mm large-size gels with five changes of solutions without destaining step; Pro-Q takes at least 250 min with 11 changes of solutions. In addition, the new method was confirmed by the study of dephosphorylation and LC-MS/MS, respectively. The approach to visualize phosphoprotein utilizing 8-Q could be an alternative to simplify the analytical operations for phosphoproteomics research.

  5. Development of a low-cost, high-throughput native polyacrylamide gel electrophoresis (N-PAGE) protocol for lipoprotein sub-fractionation using Quality by Design approach.

    PubMed

    Pathak, Mili; Chaudhary, Neha; Rathore, Anurag S

    2014-04-01

    Ratio of low density to high density lipoprotein concentration is critical for normal functioning of human body. Deviation in this ratio has been linked to various diseases, many of which are fatal if not diagnosed at early stages. For example, cardiovascular diseases (CVD) have been linked to the level of low density lipoprotein (LDL). Henceforth, detection of the lipoprotein subtractions is crucial for health of an individual. To date, methods like ultracentrifugation, nuclear magnetic resonance (NMR), high performance liquid chromatography (HPLC) and gradient gel electrophoresis (GGE) have been used for separation and identification of lipoprotein types and subtypes. However, these methods are expensive, time consuming and require specialized equipments and expertise. This paper aims to propose a low-cost, high-throughput native polyacrylamide gel electrophoresis (N-PAGE) based protocol for analysis of lipoproteins. Quality by Design (QbD) based approach has been utilized. The initial screening of parameters was followed by a fractional factorial design to optimize the protocol. The lipoprotein subtractions obtained by the optimized protocol were compared with the commercially available and commonly used Lipoprint(®) Lipoprotein Subfractions Testing System from Quantimetrix. The proposed method gave comparable results to those obtained with the commercial system. The proposed method is capable of analysis of up to forty different samples in two hours at a cost of approximately 2$/sample. This is an order of magnitude better than the present cost of 265$/sample when using the commercial system. We think that the proposed method would be of particular interest to the developing and under-developed economies of the world, where this cost differential would be deemed quite significant and would make testing affordable to the majority of the population.

  6. Separation of native allophycocyanin and R-phycocyanin from marine red macroalga Polysiphonia urceolata by the polyacrylamide gel electrophoresis performed in novel buffer systems.

    PubMed

    Wang, Yu; Gong, Xueqin; Wang, Shumei; Chen, Lixue; Sun, Li

    2014-01-01

    Three buffer systems of Imidazole-Acetic acid, HEPES-Imidazole/Bis-tris and Bis-tris-HEPES-MES were designed based on the principle of discontinuous polyacrylamide gel electrophoresis (PAGE) for the native PAGE which could be performed in pH 7.0 and 6.5 in order to analyze and prepare the minor components of allophycocyanin (AP) and R-phycocyanin (R-PC) from marine red macroalga Polysiphonia urceolata. These AP and R-PC phycobiliproteins are easily denatured in alkaline environments. The obtained results demonstrated that the PAGE modes performed in the buffer systems of HEPES-Imidazole/Bis-tris and Bis-tris-HEPES-MES gave the satisfactory resolution and separation of AP and R-PC proteins. The absorption and fluorescence spectra of the AP and R-PC proteins which were prepared by the established PAGE modes proved that they maintained natural spectroscopic characteristics. The established PAGE modes may also provide useful references and selections for some other proteins that are sensitive to alkaline environments or are not effectively separated by the classical PAGE modes performed normally in alkaline buffer systems.

  7. Lithium dodecyl sulfate/polyacrylamide gel electrophoresis of thylakoid membranes at 4 degrees C: Characterizations of two additional chlorophyll a-protein complexes.

    PubMed

    Delepelaire, P; Chua, N H

    1979-01-01

    Lithium dodecyl sulfate/polyacrylamide gel electrophoresis of Chlamydomonas reinhardtii thylakoid membranes at room temperature gave two chlorophyll-protein complexes, CP I and CP II, as had been reported previously. However, when the electrophoresis was performed at 4 degrees C, there was an increase in the amount of chlorophyll associated with CP I and CP II, and in addition, three other chlorophyll-protein complexes appeared. Two of these complexes, designated CP III and CP IV, were characterized and found to be similar in their compositions. Each complex contains four to five molecules of chlorophyll a, one molecule of beta-carotene, and one polypeptide chain. The apoprotein of CP III is polypeptide 5 (M(r) 50,000) and that of CP IV is polypeptide 6 (M(r) 47,000); the two polypeptides are structurally unrelated. Chlorophyll-protein complexes similar to C. reinhardtii CP III and CP IV were also detected in higher plants (e.g., Pisum sativum). The apoproteins of the higher plant complexes are immunochemically related to those of the C. reinhardtii complexes, as shown by crossed immunoelectrophoresis. Absorption spectra of CP III and CP IV at -196 degrees C revealed a component at 682 nm. This observation, together with the previous results on photosystem II mutants [Chua, N.-H. & Bennoun, P. (1975) Proc. Natl. Acad. Sci. USA 72, 2175-2179], provides indirect evidence that CP III and CP IV may be involved in the primary photochemistry of photosystem II.

  8. Identification of new proteins in follicular fluid from mature human follicles by direct sample rehydration method of two-dimensional polyacrylamide gel electrophoresis.

    PubMed

    Lee, Han-Chul; Lee, Sang-Wha; Lee, Kyo Won; Lee, Sook-Whan; Cha, Kwang-Yul; Kim, Kye Hyun; Lee, Suman

    2005-06-01

    Human follicular fluid (HFF) includes various biologically active proteins which can affect follicle growth and oocyte fertilization. Thus far, these proteins from mature follicles in human follicular fluid have been poorly characterized. Here, two-dimensional polyacrylamide gel electrophoresis (2-DE) with matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) was used to identify new proteins in HFF. Mature follicular fluids were obtained from five females after oocyte collection during in vitro fertilization (IVF). We directly rehydrated HFF samples, obtained high-resolution 2-DE maps, and processed them for 2-DE and MALDI-MS. One hundred eighty spots were detected and 10 of these spots were identified. By the 2-DE database, six of them had been reported, as proteins already existing in HFF. Hormone sensitive lipase (HSL), Unnamed protein product 1 (UPP1), Unnamed protein product 2 (UPP2), and apolipoprotein A-IV precursor were newly detected. HSL and apolipoprotein A-IV participate in lipid metabolism. UPP1 has a homology with selenocysteine lyase. We found by RT-PCR that these genes are expressed from human primary granulosa cells. The proteins identified here may emerge as potential candidates for specific functions during folliculogenesis, hormone secretion regulation, or oocyte maturation. Further functional analysis of these proteins is necessitated to determine their biological implications.

  9. A robust new strategy for high-molecular-weight proteome research: a 2-hydroxyethyl agarose/polyacrylamide gel enhanced separation and ZnO-PMMA nanobeads assisted identification.

    PubMed

    Shen, Wenwen; Shen, Chengpin; Xiong, Huanming; Lu, Haojie; Yang, Pengyuan

    2010-09-15

    A new mass spectrometry based analysis strategy has been established here for high-molecular-weight (HMW) proteome research. First, a 2-hydroxyethyl agarose/polyacrylamide (HEAG/PAM) electrophoresis gel was designed for the first time to realize an easy-handling separation method with high spatial resolution for HMW proteins, good reproducibility and mass spectrometry-compatible silver staining. Second, ZnO-polymethyl methacrylate (ZnO-PMMA) nanobeads were applied here for enriching and desalting the peptides from the HMW proteins. Third, the peptides were analyzed by matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) with the presence of the ZnO-PMMA nanobeads, and their MS signals were enhanced markedly. The success rate of identification for HMW proteins was significantly increased due to high enriching efficiency and salt tolerance capability as well as signal enhancing capability of the ZnO-PMMA nanobeads. We believe that this analysis strategy will inspire and accelerate the HMW proteome studies.

  10. A comparative proteomic study of plasma in feline pancreatitis and pancreatic carcinoma using 2-dimensional gel electrophoresis to identify diagnostic biomarkers: A pilot study

    PubMed Central

    Meachem, Melissa D.; Snead, Elisabeth R.; Kidney, Beverly A.; Jackson, Marion L.; Dickinson, Ryan; Larson, Victoria; Simko, Elemir

    2015-01-01

    While pancreatitis is now recognized as a common ailment in cats, the diagnosis remains challenging due to discordant results and suboptimal sensitivity of ultrasound and specific feline pancreatic lipase (Spec fPL) assay. Pancreatitis also shares similar clinical features with pancreatic carcinoma, a rare but aggressive disease with a grave prognosis. The objective of this pilot study was to compare the plasma proteomes of normal healthy cats (n = 6), cats with pancreatitis (n = 6), and cats with pancreatic carcinoma (n = 6) in order to identify potential new biomarkers of feline pancreatic disease. After plasma protein separation by 2-dimensional gel electrophoresis, protein spots were detected by Coomassie Brilliant Blue G-250 staining and identified by mass spectrometry. Alpha-1-acid glycoprotein (AGP), apolipoprotein-A1 (Apo-A1), and apolipoprotein-A1 precursor (Pre Apo-A1) appeared to be differentially expressed, which suggests the presence of a systemic acute-phase response and alteration of lipid metabolism in cats with pancreatic disease. Future studies involving greater case numbers are needed in order to assess the utility of these proteins as potential biomarkers. More sensitive proteomic techniques may also be helpful in detecting significant but low-abundance proteins. PMID:26130850

  11. Cell adhesion to proteins separated by lithium dodecyl sulfate-polyacrylamide gel electrophoresis and blotted onto a polyvinylidene difluoride membrane: a new cell-blotting technique.

    PubMed

    Seshi, B

    1994-12-02

    Cell blotting, although conceptually simple, has failed to achieve wide practical application. Described here is a new cell-blotting technique which involves cell adhesion to protein bands after separation by lithium dodecyl sulfate-polyacrylamide gel electrophoresis (LDS-PAGE) and blotting onto polyvinylidene difluoride (PVDF) membrane at 4 degrees C. Cell bands adherent on PVDF are detected using hematoxylin, or propidium iodide (PI) staining followed by viewing under ultraviolet (UV) light. The technique allows quick microscopic visualization of adherent cells composing the bands, without requiring clearing of the membrane. Representative cell adhesion proteins from different sources, i.e., plant lectins (e.g., phytohemagglutinin, PHA; concanavalin A, ConA; and wheat germ agglutinin, WGA); extracellular matrix (ECM) proteins; and integral membrane proteins (e.g., recombinant soluble vascular cell adhesion molecule-1, rs VCAM-1) were tested for cell binding by the new cell-blotting technique using human lymphoid progenitor (NALM-6) and myeloid progenitor (KG1a) cell lines. Cell adhesion proteins retained their adhesion function in all cases tested. Specificity of cell binding on PVDF blot was demonstrated by inhibition of cell adhesion to WGA protein bands using an appropriate sugar, i.e., N-acetyl D-glucosamine. The cell blotting assay was comparable in sensitivity to Coomassie blue staining of protein bands. The ability to conduct protein extraction, separation and blotting at low temperature avoids thermal denaturation, thereby preserving the adhesion properties of the proteins. The electrophoretic/blotting system has unique detergent removal/protein renaturation properties and the ability to preserve functionally active adhesion protein complexes. The cell-blotting technique described is sufficiently robust for routine application in the investigation of novel cell adhesion proteins.

  12. Quantification of bovine sperm separation by a swim-up method. Relationship to sperm motility, integrity of acrosomes, sperm migration in polyacrylamide gel and fertility.

    PubMed

    Parrish, J J; Foote, R H

    1987-01-01

    The number of bovine spermatozoa separated in a swim-up procedure was quantified using an electronic cell counter. In an initial test of the swim-up procedure, non-frozen sperm samples with different ratios of live to dead cells were prepared and tested for the number of spermatozoa counted by the swim-up procedure. In ejaculates from six bulls, the number of spermatozoa swimming up was related to the number of live cells present (R2 = 0.97). Next, sperm quality of frozen-thawed semen immediately after thawing was measured at 37 C by swim-up sperm count, sperm motility, spermatozoa with an intact acrosome and migration in polyacrylamide gel and then compared with the fertility of the semen used for artificial insemination. Twenty-nine ejaculates of frozen-thawed semen from 11 bulls were evaluated. Correlations with fertility were highest on an ejaculate basis for motility (r = 0.41, P = 0.05) and for swim-up sperm count (r = 0.35, P = 0.06). On a bull basis, swim-up sperm count had the highest correlation with fertility (r = 0.59, P = 0.06). In a multiple regression model to predict male fertility that included all described measures of semen quality, a R2 value of 0.69 was obtained. This is the first report showing that the ability of spermatozoa to swim out of a more dense medium (whole milk-glycerol extender) into culture media is quantitatively related to in vivo fertility.

  13. Qualitative and quantitative changes in barley seed protein patterns during the malting process analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis with respect to malting quality.

    PubMed

    Weiss, W; Postel, W; Görg, A

    1992-01-01

    Seeds of two barley cultivars, similar in total protein content and malt extract yield but different in their final attenuation values, were malted. Samples taken at daily intervals during the malting process were extracted sequentially with Tris-HCl buffer, aqueous 2-propanol, aqueous 2-propanol containing 0.5% dithiothreitol, and 4 M urea, containing 0.5% dithiothreitol and 1% Nonidet P-40. The protein composition of these extracts was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and computer densitometry to determine whether differences observed in the rate or extent of protein modification are related to the malting quality character final attenuation. It was found that, common to both cultivars, the albumin and globulin proteins were relatively resistant to proteolysis, whereas the hordeins suffered a dramatic breakdown during malting, with the D hordein being degraded most rapidly, followed by the B and C hordeins. Besides these similarities, differences between both cultivars were observed in the relative rates of D hordein degradation, as this rate was considerably higher in the cultivar with high malting quality. Similar, but much less distinct kinetics were seen with certain B hordeins. Since a possible relationship might exist between the rate of proteolysis of the D hordeins and the character final attenuation, we analyzed a larger number of barley cultivars with different final attenuation values with a simplified technique. For the ten cultivars examined, differences during germination were again seen in the rates of modification of the D hordeins. However, significant correlations between the D hordein breakdown and final attenuation values were not obtained, so that we propose that there exists at best a loose correlation between the relative rate of proteolysis of these proteins and the malting quality character final attenuation.

  14. Protein and glycoprotein abnormalities in platelets from human Chediak-Higashi syndrome: polyacrylamide gel electrophoretic study of platelets from five patients.

    PubMed

    Ledezma, E; Apitz-Castro, R

    1985-10-01

    Polyacrylamide electrophoretic analysis of proteins and Tritium-labelled glycoproteins of the platelets from five patients with Chediak-Higashi Syndrome shows the existence of marked quantitative differences when compared to normal platelets. While the glycoprotein abnormalities are solely related to the plasma membrane, some of the abnormalities detected in the Coomasie blue pattern are probably representative of defects related to the dense bodies and the alpha-granules. Some of the abnormalities found may, in part, explain the variability of aggregatory responses described in these patients, as well as the marked tendency towards desaggregation exhibited by platelets from humans with the Chediak-Higashi Syndrome.

  15. Optimization of Quantitative Proteomics Using 2-Dimensional Difference Gel Electrophoresis to Characterize Molecular Mechanisms of Chemical Warfare Nerve Agent Exposure in the Rat Brain

    DTIC Science & Technology

    2010-11-01

    protocol was optimized for whole rat brain, and purity was assessed using various biochemical assays. For isolation of mitochondria from brain...dependent hepatotoxicity . Journal of Biological Chemistry, 2004. 279(21): p. 22092-22101. 19. Mintz, H.A., et al., Morphological and biochemical ...Difference Gel Electrophoresis to Characterize Molecular Mechanisms of Chemical Warfare Nerve Agent Exposure in the Rat Brain Heidi M. Hoard

  16. Exposures of Sus scrofa to a TASER(®) conducted electrical weapon: no effects on 2-dimensional gel electrophoresis patterns of plasma proteins.

    PubMed

    Jauchem, James R; Cerna, Cesario Z; Lim, Tiffany Y; Seaman, Ronald L

    2014-12-01

    In an earlier study, we found significant changes in red-blood-cell, leukocyte, and platelet counts, and in red-blood-cell membrane proteins, following exposures of anesthetized pigs to a conducted electrical weapon. In the current study, we examined potential changes in plasma proteins [analyzed via two-dimensional gel electrophoresis (2-DGE)] following two 30 s exposures of anesthetized pigs (Sus scrofa) to a TASER (®) C2 conducted electrical weapon. Patterns of proteins, separated by 2-DGE, were consistent and reproducible between animals and between times of sampling. We determined that the blood plasma collection, handling, storage, and processing techniques we used are suitable for swine blood. There were no statistically significant changes in plasma proteins following the conducted-electrical-weapon exposures. Overall gel patterns of fibrinogen were similar to results of other studies of both pigs and humans (in control settings, not exposed to conducted electrical weapons). The lack of significant changes in plasma proteins may be added to the body of evidence regarding relative safety of TASER C2 device exposures.

  17. Towards a proteomic analysis of atopic dermatitis: a two-dimensional-polyacrylamide gel electrophoresis/mass spectrometric analysis of cultured patient-derived fibroblasts.

    PubMed

    Park, Yong-Doo; Kim, So-Yeon; Jang, Hee-Sun; Seo, Eun-Young; Namkung, Jung-Hyun; Park, Hyung-Seok; Cho, Sang Yun; Paik, Young-Ki; Yang, Jun-Mo

    2004-11-01

    Atopic dermatitis (AD) is a chronic relapsing inflammatory skin disease typically characterized by a distribution of eczematous skin lesions with lichenification, pruritic excoriations, and dry skin with wide varieties of pathophysiologic aspects. Recently, AD was divided into extrinsic and intrinsic forms according to the presence or absence of an allergy. We investigated alterations in protein expression in primary cultured AD cells from the patient biopsy samples by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization-time of flight. In the primary cultured fibroblasts, we obtained 31 candidate proteins from the two-dimensional gel image analysis in which 18 proteins were up-regulated, eight proteins were down-regulated and five proteins were post-translationally modified. From these 2-DE results, we found several candidate genes matched proteomic expression patterns by semiquantitative reverse transcription PCR. Since the exact mechanism of atopic alterations in fibroblasts remains unknown, our results may provide new clues to aid in understanding AD.

  18. Fluorescent staining of glycoproteins in sodium dodecyl sulfate polyacrylamide gels by 4H-[1]-benzopyrano[4,3-b]thiophene-2-carboxylic acid hydrazide.

    PubMed

    Zhu, Zhongxin; Zhou, Xuan; Wang, Yang; Chi, Lisha; Ruan, Dandan; Xuan, Yuanhu; Cong, Weitao; Jin, Litai

    2014-06-07

    A fluorescent detection method for glycoproteins in SDS-PAGE by using 4H-[1]-benzopyrano[4,3-b]thiophene-2-carboxylic acid hydrazide (BH) was developed in this study. As low as 4-8 ng glycoproteins (transferrin, α1-acid glycoprotein) could be specifically detected by the BH staining method, which is twofold more sensitive than that of the most commonly used Pro-Q Emerald 488 glycoprotein stain. Furthermore, the specificity of the newly developed stain for glycoproteins was demonstrated by 1-D and 2-D SDS-PAGE, deglycosylation, glycoprotein affinity enrichment and LC-MS/MS, respectively. According to the results, it is concluded that BH stain may provide new choices for convenient, sensitive, specific and economic visualization of gel-separated glycoproteins.

  19. Speciation analysis of inorganic arsenic in river water by Amberlite IRA 910 resin immobilized in a polyacrylamide gel as a selective binding agent for As(V) in diffusive gradient thin film technique.

    PubMed

    Rolisola, Ana M C M; Suárez, Carlos A; Menegário, Amauri A; Gastmans, Didier; Kiang, Chang H; Colaço, Camila D; Garcez, Daniel L; Santelli, Ricardo E

    2014-09-07

    In this study, a method is proposed for the selective retention of As(V) using diffusive gradient in thin film (DGT) samplers containing a strongly basic anion exchange resin (Amberlite IRA 910) supported on a polyacrylamide gel. In addition, the total arsenic content is determined by ferrihydrite gel discs. Subsequently, the concentration of As(III) was obtained by determining the difference between the total As and As(V). DGT experiments showed linear accumulation of As(V) (up to 280 ng) until a deployment time of 8 h deployment (R(2) > 0.99). The retention of As(V) was appropriate (97.9-112.3%) between pH 5 and 9. For a solution with an ionic strength ranging from 0.001 to 0.05 mol L(-1), the As(V) uptake ranged from 90-120%. The proposed method was applied for the speciation of arsenic in river water. For the analysis of spiked samples collected at the Furnas stream, the recoveries of total arsenic content ranged between 103.9% and 118.8%. However, the recoveries of As(III) and As(V) were 43.3-75.2% and 147.3-153.4%, respectively. These differences were probably because of the oxidation of As(III) to As(V) during deployments. For spiked samples collected at the Ribeirão Claro, the recoveries of dissolved As(III), As(V) and As(T) were 103.1%, 108.0% and 106.3%, respectively. Thus, the DGT technique with Amberlite IRA 910 resin as the binding phase can be employed for the in situ redox speciation of inorganic arsenic.

  20. Quantitation of estrogen receptor in seventy-five specimens of breast cancer: comparison between an immunoassay (Abbott ER-EIA monoclonal) and a (3H)estradiol binding assay based on isoelectric focusing in polyacrylamide gel

    SciTech Connect

    Pousette, A.; Gustafsson, S.A.; Thoernblad, A.M.N.; Nordgren, A.; Saellstroem, J.Li.; Lindgren, A.; Sundelin, P.; Gustafsson, J.A.

    1986-08-01

    Quantitation of estrogen receptor has been performed in cytosol prepared from 75 specimens of breast cancer tissue from patients who had not received hormonal therapy. The study was performed in order to compare an immunoassay (Abbott Laboratories, North Chicago, IL) with our currently used method for estrogen receptor analysis based on isoelectric focusing of (/sup 3/H)estradiol-receptor complex in polyacrylamide gels. Using linear regression analysis, a regression coefficient (slope) of 1.30 and a correlation coefficient of 0.75 were calculated. The differences in results between the two methods are probably partly explained by the fact that the ligand-based method only measures unoccupied receptor, whereas the immunoassay detects the total amount of receptor, resulting in generally slightly higher concentrations with the latter method. However, in five of 75 specimens the ligand-based method gave a considerably higher concentration of estrogen receptor. This was most probably explained by partial proteolysis resulting in the formation of receptor fragment(s), which was undetectable with the immunoassay but detectable with the ligand-based method. These observations underline the importance of careful handling of specimens during the whole immunoassay procedure.

  1. Gene expression in the pulp of ripening bananas. Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of in vitro translation products and cDNA cloning of 25 different ripening-related mRNAs.

    PubMed Central

    Medina-Suárez, R; Manning, K; Fletcher, J; Aked, J; Bird, C R; Seymour, G B

    1997-01-01

    mRNA was extracted from the pulp and peel of preclimacteric (d 0) bananas (Musa AAA group, cv Grand Nain) and those exposed to ethylene gas for 24 h and stored in air alone for a further 1 (d 2) and 4 d (d 5). Two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis of in vitro translation products from the pulp and peel of these fruits revealed significant up-regulation of numerous transcripts during ripening. The majority of the changes were initiated by d 2, with the level of these messages increasing during the remainder of the ripening period. Pulp tissue from d 2 was used for the construction of a cDNA library. This library was differentially screened for ripening-related clones using cDNA from d-0 and d-2 pulp by a novel microtiter plate method. In the primary screen 250 up- and down-regulated clones were isolated. Of these, 59 differentially expressed clones were obtained from the secondary screen. All of these cDNAs were partially sequenced and grouped into families after database searches. Twenty-five nonredundant groups of pulp clones were identified. These encoded enzymes were involved in ethylene biosynthesis, respiration, starch metabolism, cell wall degradation, and several other key metabolic events. We describe the analysis of these clones and their possible involvement in ripening. PMID:9342865

  2. Direct assay for O6-methylguanine-DNA methyltransferase and comparison of detection methods for the methylated enzyme in polyacrylamide gels and electroblots.

    PubMed

    Major, G N; Gardner, E J; Lawley, P D

    1991-07-01

    other human cells and tissues, extracts of human spleen in the present study showed wide interindividual differences in O6-MT specific activity (18-fold), which spanned the range 50-900 fmol/mg of protein. Cultured human lymphoblastoid Jurkat cells contained approx. 57,000 enzyme molecules/cell. Substrate-inactivated [Me-3H]methylated O6-MT was analysed by SDS/PAGE and electroblotting. The different but similarly sized forms of this enzyme that we previously detected in human spleen [Major, Gardner, Carne & Lawley (1990) Nucleic Acids Res. 18, 1351-1359] were clearly resolved by fluorography of electroblots, but only at considerable expense of time. As expected, scintillation counting of the protein extracted from gel slices and linear-wire scanning of enzyme-associated radioactivity on electroblots were quicker methods for detecting the [Me-3H]methylated inactivated O6-MT.

  3. Changes in protein abundance between tender and tough meat from bovine longissimus thoracis muscle assessed by isobaric Tag for Relative and Absolute Quantitation (iTRAQ) and 2-dimensional gel electrophoresis analysis.

    PubMed

    Bjarnadóttir, S G; Hollung, K; Høy, M; Bendixen, E; Codrea, M C; Veiseth-Kent, E

    2012-06-01

    The aim of this study was to find potential biomarkers for meat tenderness in bovine Longissimus thoracis muscle and to compare results from isobaric Tag for Relative and Absolute Quantitation (iTRAQ) and 2-dimensional gel electrophoresis (2-DE) analysis. The experiment included 4 tender and 4 tough samples, based on shear force measurements at 7 d postmortem, from young Norwegian red (NRF) bulls, taken at 1 h postmortem. A number of the proteins which have previously been related to tenderness were found to change in abundance between tender and tough samples, both in iTRAQ (P < 0.1) and 2-DE analysis (P < 0.05). Furthermore, 3 proteins that have not previously been related to tenderness were found to change significantly in abundance between tender and tough meat samples in the present study. These include proteins related to control of flux through the tricarboxylate cycle [2-oxoglutarate dehydrogenase complex component E2 (OGDC-E2)], apoptosis (galectin-1) and regulatory role in the release of Ca(2+) from intracellular stores (annexin A6). Even though the overlap in significantly changing proteins was relatively low between iTRAQ and 2-DE analysis, certain proteins predicted to have the same function were found in both analyses and showed similar changes between the groups, such as structural proteins and proteins related to apoptosis and energy metabolism.

  4. 21 CFR 172.255 - Polyacrylamide.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ..., Films and Related Substances § 172.255 Polyacrylamide. Polyacrylamide containing not more than 0.2 percent of acrylamide monomer may be safely used as a film former in the imprinting of soft-shell...

  5. 21 CFR 172.255 - Polyacrylamide.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ..., Films and Related Substances § 172.255 Polyacrylamide. Polyacrylamide containing not more than 0.2 percent of acrylamide monomer may be safely used as a film former in the imprinting of soft-shell...

  6. 21 CFR 172.255 - Polyacrylamide.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ..., Films and Related Substances § 172.255 Polyacrylamide. Polyacrylamide containing not more than 0.2 percent of acrylamide monomer may be safely used as a film former in the imprinting of soft-shell...

  7. 21 CFR 172.255 - Polyacrylamide.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ..., Films and Related Substances § 172.255 Polyacrylamide. Polyacrylamide containing not more than 0.2 percent of acrylamide monomer may be safely used as a film former in the imprinting of soft-shell...

  8. Nanotube-grafted polyacrylamide hydrogels for electrophoretic protein separation.

    PubMed

    Gunavadhi, Murugappan; Maria, Lourdusamy Arul Antony; Chamundeswari, Vidya N; Parthasarathy, Meera

    2012-04-01

    Multiwalled carbon nanotube-modified polyacrylamide gels have been employed for the electrophoretic separation of proteins. Two approaches are compared in this investigation, one using nanotubes only as fillers inside the gel matrix and the other using nanotubes as catalyst for polymerization of acrylamide. In both the cases, polymerization of acryl-amide/bisacrylamide has been carried out in situ in the presence of nanotubes dispersed in the gel buffer containing monomer and cross-linker. In the former case, initiator and catalyst have been added after ultrasonication of nanotubes in the gel buffer mixture where the nanotubes play the role of filler. On the other hand, the second approach precludes use of catalyst and involves addition of initiator alone during ultrasonication of nanotubes in the gel buffer containing monomer and cross-linker, which leads to the formation of nanotube-grafted gel after 25 min. When nanotubes are used as a catalyst instead of N,N,N',N'-tetramethylethylenediamine, pore size distribution of the gel matrix and linearity of molecular weight calibration plots are found to be improved. In addition, other issues associated with the use of an external catalyst like handling the moisture-sensitive and corrosive reagent and associated irreproducibility are addressed in this approach.

  9. Network generation enhances interpretation of proteomics data sets by a combination of two-dimensional polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization-time of flight mass spectrometry.

    PubMed

    Wang, Xijun; Zhang, Aihua; Sun, Hui; Wu, Gelin; Sun, Wenjun; Yan, Guangli

    2012-10-21

    Recent advances in proteomic technologies have enabled us to create detailed protein-protein interaction maps in diseases. As the size of the interaction dataset increases, powerful computational methods are required in order to effectively interpret network models from large scale interactome data. In this study, we carried out comparative proteomics to construct and identify the proteins networks associated with hepatic injury (HI) which are largely unknown, as a case study. All proteins expressed were separated and identified by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight-time-of-flight mass spectrometry (MALDI-TOF/TOF MS). Protein-interacting networks and pathways were mapped using STRING analysis program. We have performed for the first time a comprehensive profiling of changes in protein expression of HI rats, to uncover the networks altered by treated with CCl(4). Identification of fifteen spots (seven over-expressed and eight under-expressed) were established by MALDI-TOF/TOF MS. These proteins were subjected to functional pathway analysis using STRING software for better understanding of the biological context of the identified proteins. It suggested that modulation of multiple vital physiological pathways including DNA repair process, cell apoptosis, oxidation reduction, signal transduction, metabolic process, intracellular signaling cascade, regulation of biological processes, cell communication, regulation of cellular process, and molecular transport. In summary, the present study provides the first protein-interacting network maps and novel insights into the biological responses and potential pathways of HI. The generation of protein interaction networks clearly enhances the interpretation of proteomic data, particularly in respect of understanding molecular mechanisms of panel protein biomarkers.

  10. Spring-loaded polymeric gel actuators

    DOEpatents

    Shahinpoor, M.

    1995-02-14

    Spring-loaded electrically controllable polymeric gel actuators are disclosed. The polymeric gels can be polyvinyl alcohol, polyacrylic acid, or polyacrylamide, and are contained in an electrolytic solvent bath such as water plus acetone. The action of the gel is mechanically biased, allowing the expansive and contractile forces to be optimized for specific applications. 5 figs.

  11. Spring-loaded polymeric gel actuators

    DOEpatents

    Shahinpoor, Mohsen

    1995-01-01

    Spring-loaded electrically controllable polymeric gel actuators are disclosed. The polymeric gels can be polyvinyl alcohol, polyacrylic acid, or polyacrylamide, and are contained in an electrolytic solvent bath such as water plus acetone. The action of the gel is mechanically biased, allowing the expansive and contractile forces to be optimized for specific applications.

  12. An overlay gel method for identification and isolation of bacterial beta-lactamases.

    PubMed

    Eftekhar, Fereshteh; Rafiee, Roya

    2006-01-01

    A modification of the iodometric technique using an overlay gel was employed for fast identification and isolation of beta-lactamase types TEM, SHV and AmpC from non-denaturing gels. Osmotic shock preparations of the three beta-lactamases were run on polyacrylamide gels without SDS and ampicillin containing overlay gels were flooded with the iodine solution before being placed on polyacrylamide gel strips. Distinct clear bands appeared in dark blue backgrounds indicating beta-lactamase activity.

  13. Evaluation du potentiel radiosensibilisateur ou radioprotecteur/antioxydant de quelques composes selectionnes par dosimetrie par gel de polyacrylamide et dosimetre de Fricke, et utilisation de la filamentation par impulsion laser infrarouge fenitoseconde comme un nouveau et puissant faisceau pour la radiotherapie du cancer

    NASA Astrophysics Data System (ADS)

    Meesat, Ridthee

    In radiation treatment, a sufficiently high radiation dose must be delivered to the tissue volumes containing the tumor cells while the lowest possible dose should be deposited in surrounding healthy tissue. We developed an original approach that is fast and easy to implement for the early assessment of the efficiency of radiation sensitizers and protectors. In addition, we characterized a new femtosecond laser pulse irradiation technique. We are able to deposit a considerable dose with a very high dose rate inside a well-controlled macroscopic volume without deposition of energy in front or behind the target volume. The radioprotective efficiency was measured by irradiation of the Fricke solution incorporating a compound under study and measuring the corresponding production of ferric ions G(Fe3+). The production of ferric ions is most sensitive to the radical species produced in the radiolysis of water. We studied experimentally and simulated with a full Monte-Carlo computer code the radiation-induced chemistry of Fricke/cystamine solutions. Results clearly indicate that the protective effect of cystamine originates from its radical-capturing ability, which allows this compound to compete with the ferrous ions for the various free radicals - especially ·OH radicals and H· atoms - formed during irradiation of the surrounding water. The sensitizing capacity of radiation sensitizers was measured by irradiation of a polyacrylamide gel (PAG) dosimeter incorporating a compound under study and measuring the corresponding increase in the gradient between spin-spin relaxation rate (R2) and absorbed dose. We measured an irradiation energy-dependent increase in R 2-dose sensitivity for halogenated compounds or a decrease for radioprotectors. Finally, we studied a novel laser irradiation method called "filamentation". We showed that this phenomenon results in an unprecedented deposition of energy and the dose rate thus achieved exceeds by orders of magnitude values

  14. Crystallization of calcium phosphate in polyacrylamide hydrogels containing phosphate ions

    NASA Astrophysics Data System (ADS)

    Yokoi, Taishi; Kawashita, Masakazu; Kikuta, Koichi; Ohtsuki, Chikara

    2010-08-01

    Calcium phosphate crystals were formed in polyacrylamide (PAAm) hydrogels containing phosphate ions by diffusion of calcium ions from calcium nitrate (Ca(NO 3) 2) solutions covering the gels. Changes in crystalline phases and crystal morphology of calcium phosphate, and in ion concentrations of the Ca(NO 3) 2 solutions were investigated as a function of reaction time. Single or two coexisting crystalline phases of calcium phosphate, hydroxyapatite (HAp), HAp/dicalcium phosphate dihydrate (DCPD) or octacalcium phosphate (OCP)/DCPD were formed in the gels. HAp crystals are formed near the surface of the gels. The dense HAp layer and HAp/DCPD layer prevented diffusion of calcium ions from the Ca(NO 3) 2 solution, thus formation of calcium phosphate in the gel phase was inhibited. Formation of DCPD was observed to follow the formation of OCP or HAp. The size of the OCP crystals gradually increased with reaction time, while changes in size of HAp crystals were not observed. The reaction time required for DCPD formation depended on the degree of supersaturation with respect to DCPD in the systems. DCPD formed within 1 day under high supersaturation conditions, whereas it formed at 10 days in low supersaturation conditions.

  15. A composite agarose-polyacrylamide matrix as two-dimensional hard support for solid-phase protein assays.

    PubMed

    Krajewski, Wladyslaw A

    2016-03-15

    The solid-phase protein assays using blotting membranes as hard support do not allow achieving the low background and sensitivity of protein staining in clear gels. The membrane opacity complicates imaging of results on standard lab documentation systems. We describe a low-cost transparent matrix that can be used as an alternative to polymeric membranes for solid-phase assays. Protein samples are spotted onto a dry film of composite agarose-polyacrylamide matrix covering standard glass microscopic slides. After rehydration in protein-fixing solution, matrix with protein samples can be detached from glass support and stained as conventional protein polyacrylamide gels.

  16. Electrically controlled polymeric gel actuators

    DOEpatents

    Adolf, Douglas B.; Shahinpoor, Mohsen; Segalman, Daniel J.; Witkowski, Walter R.

    1993-01-01

    Electrically controlled polymeric gel actuators or synthetic muscles capable of undergoing substantial expansion and contraction when subjected to changing pH environments, temperature, or solvent. The actuators employ compliant containers for the gels and their solvents. The gels employed may be cylindrical electromechanical gel fibers such as polyacrylamide fibers or a mixture of poly vinyl alcohol-polyacrylic acid arranged in a parallel aggregate and contained in an electrolytic solvent bath such as salt water. The invention includes smart, electrically activated devices exploiting this phenomenon. These devices are capable of being manipulated via active computer control as large displacement actuators for use in adaptive structure such as robots.

  17. Electrically controlled polymeric gel actuators

    DOEpatents

    Adolf, D.B.; Shahinpoor, M.; Segalman, D.J.; Witkowski, W.R.

    1993-10-05

    Electrically controlled polymeric gel actuators or synthetic muscles are described capable of undergoing substantial expansion and contraction when subjected to changing pH environments, temperature, or solvent. The actuators employ compliant containers for the gels and their solvents. The gels employed may be cylindrical electromechanical gel fibers such as polyacrylamide fibers or a mixture of poly vinyl alcohol-polyacrylic acid arranged in a parallel aggregate and contained in an electrolytic solvent bath such as salt water. The invention includes smart, electrically activated devices exploiting this phenomenon. These devices are capable of being manipulated via active computer control as large displacement actuators for use in adaptive structure such as robots. 11 figures.

  18. Bioconversion of acrylonitrile to acrylamide using polyacrylamide entrapped cells of Rhodococcus rhodochrous PA-34.

    PubMed

    Raj, J; Prasad, S; Sharma, N N; Bhalla, T C

    2010-09-01

    The nitrile hydratase (NHase) of Rhodococcus rhodochrous PA-34 catalyzed the conversion of acrylonitrile to acrylamide. The resting cells (having NHase activity) (8 %; 1 mL corresponds to 22 mg dry cell mass, DCM) were immobilized in polyacrylamide gel containing 12.5 % acrylamide, 0.6 % bisacrylamide, 0.2 % diammonium persulfate and 0.4 % TEMED. The polyacrylamide entrapped cells (1.12 mg DCM/mL) completely converted acrylonitrile in 3 h at 10 °C, using 0.1 mol/L potassium phosphate buffer. In a partitioned fed batch reactor, 432 g/L acrylamide was accumulated after 1 d. The polyacrylamide discs were recycled up to 3×; 405, 210 and 170 g/L acrylamide was produced in 1st, 2nd and 3rd recycling reactions. In four cycles, a total of 1217 g acrylamide was produced by recycling the same mass of entrapped cells.

  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. Polyacrylamide Hydrogel Properties for Horticultural Applications

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Polyacrylamide (PAAm) hydrogels are commonly employed to ensure hydration of the growth media and minimize crop losses during the crop production and postproduction phases in horticulture. However, studies of the effect of these materials have shown that they have a minimal effect on crop life and q...

  1. 21 CFR 173.10 - Modified polyacrylamide resin.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... conditions: (a) The modified polyacrylamide resin is produced by the copolymerization of acrylamide with not... polyacrylamide resin contains not more than 0.05 percent residual acrylamide. (c) The modified...

  2. 21 CFR 173.10 - Modified polyacrylamide resin.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... conditions: (a) The modified polyacrylamide resin is produced by the copolymerization of acrylamide with not... polyacrylamide resin contains not more than 0.05 percent residual acrylamide. (c) The modified...

  3. 21 CFR 173.10 - Modified polyacrylamide resin.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... conditions: (a) The modified polyacrylamide resin is produced by the copolymerization of acrylamide with not... polyacrylamide resin contains not more than 0.05 percent residual acrylamide. (c) The modified...

  4. 21 CFR 173.10 - Modified polyacrylamide resin.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... conditions: (a) The modified polyacrylamide resin is produced by the copolymerization of acrylamide with not... polyacrylamide resin contains not more than 0.05 percent residual acrylamide. (c) The modified...

  5. 21 CFR 173.10 - Modified polyacrylamide resin.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... polyacrylamide resin is produced by the copolymerization of acrylamide with not more than 5-mole percent β....05 percent residual acrylamide. (c) The modified polyacrylamide resin is used as a flocculent in...

  6. Formation of composite polyacrylamide and silicone substrates for independent control of stiffness and strain.

    PubMed

    Simmons, Chelsey S; Ribeiro, Alexandre J S; Pruitt, Beth L

    2013-02-21

    Cells that line major tissues in the body such as blood vessels, lungs and gastrointestinal tract experience deformation from mechanical strain with our heartbeat, breathing, and other daily activities. Tissues also remodel in both development and disease, changing their mechanical properties. Taken together, cells can experience vastly different mechanical cues resulting from the combination of these interdependent stimuli. To date, most studies of cellular mechanotransduction have been limited to assays in which variations in substrate stiffness and strain were not combined. Here, we address this technological gap by implementing a method that can simultaneously tune both substrate stiffness and mechanical strain. Substrate stiffness is controlled with different monomer and crosslinker ratios during polyacrylamide gel polymerization, and strain is transferred from the underlying silicone platform when stretched. We demonstrate this platform with polyacrylamide gels with elastic moduli at 6 kPa and 20 kPa in combination with two different silicone formulations. The gels remain attached with up to 50% applied strains. To validate strain transfer through the gels into cells, we employ particle-tracking methods and observe strain transmission via cell morphological changes.

  7. Mechanical Properties of DNA-Crosslinked Polyacrylamide Hydrogels with Increasing Crosslinker Density.

    PubMed

    Previtera, Michelle L; Chippada, Uday; Schloss, Rene S; Yurke, Bernard; Langrana, Noshir A

    2012-10-01

    DNA-cross-linked polyacrylamide hydrogels (DNA gels) are dynamic mechanical substrates. The addition of DNA oligomers can either increase or decrease the crosslinker density to modulate mechanical properties. These DNA-responsive gels show promise as substrates for cell culture and tissue-engineering applications, since the gels allow time-dependent mechanical modulation. Previously, we reported that fibroblasts plated on DNA gels responded to modulation in elasticity via an increase or decrease in crosslinker density. To better characterize fibroblast mechanical signals, changes in stress and elastic modulus of DNA gels were measured over time as crosslinker density altered. In a previous study, we observed that as crosslinker density decreased, stress was generated, and elasticity changed over time; however, we had not evaluated stress and elastic modulus measurements of DNA gels as crosslinker density increased. Here, we completed this set of fibroblast studies by reporting stress and elastic modulus measurements over time as the crosslinker density increased. We found that the stress generated and the elastic modulus alterations were correlated. Hence, it seemed impossible to separate the effect of stress from the effect of modulus changes for fibroblasts plated on DNA gels. Yet, previous results and controls revealed that stress contributed to fibroblast behavior.

  8. Conducting polymer electrodes for gel electrophoresis.

    PubMed

    Bengtsson, Katarina; Nilsson, Sara; Robinson, Nathaniel D

    2014-01-01

    In nearly all cases, electrophoresis in gels is driven via the electrolysis of water at the electrodes, where the process consumes water and produces electrochemical by-products. We have previously demonstrated that π-conjugated polymers such as poly(3,4-ethylenedioxythiophene) (PEDOT) can be placed between traditional metal electrodes and an electrolyte to mitigate electrolysis in liquid (capillary electroosmosis/electrophoresis) systems. In this report, we extend our previous result to gel electrophoresis, and show that electrodes containing PEDOT can be used with a commercial polyacrylamide gel electrophoresis system with minimal impact to the resulting gel image or the ionic transport measured during a separation.

  9. Evaporation of 2-Dimensional Black Holes

    NASA Astrophysics Data System (ADS)

    Ramazanoglu, Fethi M.

    2011-04-01

    Violation of unitarity in black hole evaporation has been puzzling physicist since the seminal work of Hawking in the seventies. Although there are hopes for a resolution of the problem in a full theory of quantum gravity, it has eluded us so far. Even less ambitious efforts considering only quantum corrections beyond the external field approximation have proven hard to attack in 4 dimensions. All these obstacles directed researchers to investigate the black hole evaporation problem in simpler 2-dimensional models. In this talk, we will present results on a new investigation of one of these models, the 2-dimensional Callan-Giddings-Harvey-Strominger (CGHS) model. Using a combination of analytical and high precision numerical tools, we are able to resolve CGHS black hole evaporation within the mean field approximation all the way to the point where the black hole area vanishes. Our results confirm some of the assumptions of the standard paradigm, and strongly suggest the recovery of unitarity within the full quantum theory. On the other hand, there are several surprising new results, in particular remarkable universal behavior in the evaporation of initially macroscopic black holes. This suggests that information about the collapsing matter that formed the black hole can not be recovered from the evaporation radiation. Though this separation of the questions of information loss and unitarity is peculiar to the 2-dimensional model, insights into the higher dimensional case can still be garnered.

  10. Quantification of viscoelastic effects of polyacrylamide solutions

    SciTech Connect

    Heemskerk, J.; Holtslag, R.J.; Janssen-van Rosmalen, R.; Teeuw, D.

    1984-04-01

    Beyond a critical rate of flow in cores the viscoelasticity of polyacrylamide solutions is reflected by shear thickening behaviour. Practical information is provided on the critical flow rate and magnitude of shear thickening and their dependency on core permeability, temperature, molecular weight and concentration of commercial polymers. To describe the onset of shear thickening, critical Deborah numbers were calculated using fluid relaxation times obtained by oscillation rheometry. Using three core-flow parameters, viz. the critical flow rate and two powerlaw exponents, the viscoelastic effects are quantified.

  11. Purification of radiolabeled RNA products using denaturing gel electrophoresis

    PubMed Central

    Adachi, Hironori; Yu, Yi-Tao

    2014-01-01

    This unit discusses a basic method for purification of radiolabeled RNAs using denaturing polyacrylamide gel electrophoresis. The method consists of a number of experimental procedures, including total RNA preparation from yeast cells, isolation of a specific RNA from total yeast RNA, RNA 3' terminal labeling using nucleotide (5’[32P]pCp) addition (via ligation), denaturing (8 M urea) polyacrylamide gel electrophoresis, and RNA extraction from the gel slice. Key points for achieving good electrophoretic separation of RNA are also discussed. PMID:24510465

  12. Radio-synthesized polyacrylamide hydrogels for proteins release

    NASA Astrophysics Data System (ADS)

    Ferraz, Caroline C.; Varca, Gustavo H. C.; Lopes, Patricia S.; Mathor, Monica B.; Lugão, Ademar B.

    2014-01-01

    The use of hydrogels for biomedical purposes has been extensively investigated. Pharmaceutical proteins correspond to highly active substances which may be applied for distinct purposes. This work concerns the development of radio-synthesized hydrogel for protein release, using papain and bovine serum albumin as model proteins. The polymer was solubilized (1% w/v) in water and lyophilized. The proteins were incorporated into the lyophilized polymer and the hydrogels were produced by simultaneous crosslinking and sterilization using γ-radiation under frozen conditions. The produced systems were characterized in terms of swelling degree, gel fraction, crosslinking density and evaluated according to protein release, bioactivity and cytotoxicity. The hydrogels developed presented different properties as a function of polymer concentration and the optimized results were found for the samples containing 4-5% (w/v) polyacrylamide. Protein release was controlled by the electrostatic affinity of acrylic moieties and proteins. This selection was based on the release of the proteins during the experiment period (up to 50 h), maintenance of enzyme activity and the nanostructure developed. The system was suitable for protein loading and release and according to the cytotoxic assay it was also adequate for biomedical purposes, however this method was not able to generate a matrix with controlled pore sizes.

  13. Performance and Biocompatibility of Extremely Tough Alginate/Polyacrylamide Hydrogels

    PubMed Central

    Darnell, Max; Sun, Jeong-Yun; Mehta, Manav; Johnson, Chris; Arany, Praveen; Suo, Zhigang

    2013-01-01

    Although hydrogels now see widespread use in a host of applications, low fracture toughness and brittleness have limited their more broad use. As a recently described interpenetrating network (IPN) of alginate and polyacrylamide demonstrated a fracture toughness of ∼9000 J/m2, we sought to explore the biocompatibility and maintenance of mechanical properties of these hydrogels in cell culture and in vivo conditions. These hydrogels can sustain a compressive strain of over 90% with minimal loss of Young's Modulus as well as minimal swelling for up to 50 days of soaking in culture conditions. Mouse mesenchymal stem cells exposed to the IPN gel-conditioned media maintain high viability, and although cells exposed to conditioned media demonstrate slight reductions in proliferation and metabolic activity (WST assay), these effects are abrogated in a dose-dependent manner. Implantation of these IPN hydrogels into subcutaneous tissue of rats for 8 weeks led to mild fibrotic encapsulation and minimal inflammatory response. These results suggest the further exploration of extremely tough alginate/PAAM IPN hydrogels as biomaterials. PMID:23896005

  14. Swelling kinetics of microgels embedded in a polyacrylamide hydrogel matrix.

    PubMed

    Huang, Na; Guan, Ying; Zhu, X X; Zhang, Yongjun

    2014-06-23

    Composite hydrogels--macroscopic hydrogels with embedded microgel particles--are expected to respond to external stimuli quickly because microgels swell much faster than bulky gels. In this work, the kinetics of the pH-induced swelling of a composite hydrogel are studied using turbidity measurements. The embedded microgel is a pH- and thermosensitive poly(N-isopropylacrylamide-co-acrylic acid) microgel and the hydrogel matrix is polyacrylamide. A rapid pH-induced swelling of the embedded microgel particles is observed, confirming that composite hydrogels respond faster than ordinary hydrogels. However, compared with the free microgels, the swelling of the embedded microgel is much slower. Diffusion of OH(-) into the composite hydrogel film is identified as the main reason for the slow swelling of the embedded microgel particles, as the time of the pH-induced swelling of this film is comparable to that of OH(-) diffusion into the film. The composition of the hydrogel matrix does not significantly change the characteristic swelling time of the composite hydrogel film. However, the swelling pattern of the film changes with composition of the hydrogel matrix.

  15. Diffusion of polymer gel implants.

    PubMed

    Davis, B K

    1974-08-01

    Crosslinked polyacrylamide and polyvinylpyrrolidone gels have been used to subcutaneously implant (125)I-labeled immunoglobulin, (125)I-labeled luteinizing hormone, (125)I-labeled bovine serum albumin, (125)I-labeled insulin, [(3)H]prostaglandin F(2alpha), and Na(125)I into hamsters. From the rates of absorption of the solutes, their diffusion coefficients were determined. The diffusion coefficients showed a logarithmic dependence on implant polymer concentration and solute molecular weight. Release of the solutes from gel preparations incubated 10 mM phosphate buffer (pH 7.2) at 37 degrees revealed a similar relationship between solute diffusion coefficient, molecular weight, and the concentration of polymer. A general equation was derived that gives the expected diffusion coefficient of a substance in a polymer gel from its molecular weight, diffusion coefficient in solvent, and polymer concentration of the gel.

  16. A simple and cost-effective solid-phase protein nano-assay using polyacrylamide-coated glass plates.

    PubMed

    Krajewski, Wladyslaw A

    2015-02-01

    A new solid-phase protein nano-assay is suggested for simple and sensitive estimation of protein content in sample buffers (a 1-μl sample is sufficient for analysis). The assay is different from conventional "on-filter" assays in that it uses inexpensive fully transparent polyacrylamide gel (PAAG)-coated glass plates as solid support and, thus, combines the convenience of "on-membrane" staining with the sensitivity and ease of documentation of "in-gel" staining (and, therefore, is especially suited for standard lab gel documentation systems). The PAAG plates assay is compatible with all dyes for in-gel protein staining. Depending on the sensitivity of the staining protocol, the assay can be used in macro-, micro-, and nano-assay formats. We also describe a low-cost two-component colloidal Coomassie brilliant blue G-250 (CBB G-250) staining protocol for fast quantitative visualization of proteins spotted on a PAAG plate (the detection limit is up to 2 ng of proteins even when using a Nikon CoolPix digital camera and white light transilluminator instead of a gel scanner). The suggested colloidal CBB G-250 protocol could also be used for visualizing nano-amounts of proteins in polyacrylamide gels. The PAAG plate assay could be useful for proteomic applications and, in general, for all cases where a fast, sensitive, and easily documentable cost-effective solid-phase protein assay is required.

  17. Data of microstructure and mechanical properties of carbon foams derived from sucrose/polyacrylamide hydrogel

    PubMed Central

    Yao, Yao; Chen, Fei; Chen, Xi; Shen, Qiang; Zhang, Lianmeng

    2016-01-01

    An easy method that combined gel casting and physical foaming was used to fabricate modified carbon foams. The design of carbon foams from sucrose/polyacrylamide hydrogel is a new concept for controlling the microstructure and improving the compressive properties of carbon foams. This article provides the micrographs obtained from optical and scanning electron microscope for foaming solution and carbon foams. Weight loss data used to construct the thermo-gravimetric curves are included. Load–displacement data constructing the stress–strain curves and the derived compressive properties are also included. PMID:26933668

  18. 21 CFR 872.3420 - Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... polyacrylamide polymer denture adhesive. 872.3420 Section 872.3420 Food and Drugs FOOD AND DRUG ADMINISTRATION....3420 Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive. (a) Identification. A carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive is a...

  19. 21 CFR 872.3420 - Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... polyacrylamide polymer denture adhesive. 872.3420 Section 872.3420 Food and Drugs FOOD AND DRUG ADMINISTRATION....3420 Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive. (a) Identification. A carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive is a...

  20. 21 CFR 872.3420 - Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... polyacrylamide polymer denture adhesive. 872.3420 Section 872.3420 Food and Drugs FOOD AND DRUG ADMINISTRATION....3420 Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive. (a) Identification. A carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive is a...

  1. 21 CFR 872.3420 - Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... polyacrylamide polymer denture adhesive. 872.3420 Section 872.3420 Food and Drugs FOOD AND DRUG ADMINISTRATION....3420 Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive. (a) Identification. A carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive is a...

  2. 21 CFR 872.3420 - Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... polyacrylamide polymer denture adhesive. 872.3420 Section 872.3420 Food and Drugs FOOD AND DRUG ADMINISTRATION....3420 Carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive. (a) Identification. A carboxymethylcellulose sodium and cationic polyacrylamide polymer denture adhesive is a...

  3. Spectrophotometric determination of substrate-borne polyacrylamide.

    PubMed

    Lu, Jianhang; Wu, Laosheng

    2002-08-28

    Polyacrylamides (PAMs) have wide application in many industries and in agriculture. Scientific research and industrial applications manifested a need for a method that can quantify substrate-borne PAM. The N-bromination method (a PAM analytical technique based on N-bromination of amide groups and spectrophotometric determination of the formed starch-triiodide complex), which was originally developed for determining PAM in aqueous solutions, was modified to quantify substrate-borne PAM. In the modified method, the quantity of substrate-borne PAM was converted to a concentration of starch-triiodide complex in aqueous solution that was then measured by spectrophotometry. The method sensitivity varied with substrates due to sorption of reagents and reaction intermediates on the substrates. Therefore, separate calibration for each substrate was required. Results from PAM samples in sand, cellulose, organic matter burnt soils, and clay minerals showed that this method had good accuracy and reproducibility. The PAM recoveries ranged from 95.8% to 103.7%, and the relative standard deviations (n = 4) were <7.5% in all cases. The optimum range of PAM in each sample is 10-80 microg. The technique can serve as an effective tool in improving PAM application and facilitating PAM-related research.

  4. Field demonstration of in situ grouting of radioactive solid waste burial trenches with polyacrylamide. [Polyacrylamide

    SciTech Connect

    Spalding, B.P.; Fontaine, T.A.

    1990-01-01

    Demonstrations of in situ grouting with polyacrylamide were carried out on two undisturbed burial trenches and one dynamically compacted burial trench in Solid Waste Storage Area (SWSA) 6 at Oak Ridge National Laboratory (ORNL). The injection of polyacrylamide was achieved quite facilely for the two undisturbed burial trenches which were filled with grout, at typical pumping rates of 95 L/min, in several batches injected over several days. The compacted burial trench, however, failed to accept grout at more than 1.9 L/min even when pressure was applied. Thus, it appears that burial trenches, stabilized by dynamic compaction, have a permeability too low to be considered groutable. The water table beneath the burial trenches did not respond to grout injections indicating a lack of hydrologic connection between fluid grout and the water table which would have been observed if the grout failed to set. Because grout set times were adjusted to less than 60 min, the lack of hydrologic connection was not surprising. Postgrouting penetration testing revealed that the stability of the burial trenches was increased from 26% to 79% that measured in the undisturbed soil surrounding the trenches. In situ permeation tests on the grouted trenches indicated a significant reduction in hydraulic conductivity of the trench contents from a mean of 2.1 {times} 10{sup {minus}3} to 1.85 {times} 10{sup {minus}5} cm/s. Preliminary observations indicated that grouting with polyacrylamide is an excellent method for both improved stability and hydrologic isolation of radioactive waste and its incidental hazardous constituents.

  5. BANANA GEL.

    PubMed

    McGuire, G; Falk, K G

    1922-03-20

    The conditions for the formation of gels from banana extracts were studied. Gels were obtained with extracts more alkaline than pH 7.0 with very small quantities of calcium, strontium, and barium salts, the gel formation with these salts decreasing in the indicated order. In solutions more acid than pH 6.0, no gels were obtained with these salts. Magnesium, lithium, and sodium salts did not cause gel formation either in acid or alkaline solutions. Pancreatine gave a gel on incubation with banana extract at pH 5.0. The gel-forming property of banana extracts was destroyed on boiling.

  6. In situ grouting of buried transuranic waste with polyacrylamide

    SciTech Connect

    Spalding, B.P.; Lee, S.Y.; Farmer, C.D.; Hyder, L.K.; Supaokit, P.

    1987-01-01

    This project is a demonstration and evaluation of the in situ hydrologic stabilization of buried transuranic waste at a humid site via grout injection. Two small trenches, containing buried transuranic waste, were filled with 34.000 L of polyacrylamide grout. Initial field results have indicated that voids within the trenches were totally filled by the grout and that the intratrench hydraulic conductivity was reduced to below field-measurable values. No evidence of grout constituents were observed in twelve perimeter groundwater monitoring wells indicating that grout was contained completely within the two trenches. Polyacrylamide grout was selected for field demonstration over the polyacrylate grout due to its superior performance in laboratory degradation studies. Also supporting the selection of polyacrylamide was the difficulty in controlling the set time of the acrylate polymerization. Based on preliminary degradation monitoring, the polyacrylamide was estimated to have a microbiological half-life of 362 years in the test soil. 15 refs., 9 figs., 12 tabs.

  7. Polyacrylamide Transport in Water Delivery Canals

    NASA Astrophysics Data System (ADS)

    Chen, L.; Zhu, J.; Young, M.

    2007-12-01

    Linear, anionic polyacrylamide (PAM) is being considered in the western United States as a technology to reduce seepage in unlined water delivery canals. A broad laboratory and field testing program has been undertaken to understand the benefits and potential environmental impacts of PAM use. The ability to predict the fate and transport of PAM in water delivery canals could prove to be a useful planning tool for PAM application. However, one key area of uncertainty of this type of canal treatment is the hydration, reaction, and settling rates of PAM after the dry powder is added to the canal water. In this study, we have developed a model that incorporates a number of known physical and chemical processes that can affect PAM transport, such as convection, dispersion, dissolution, flocculation, and settling, while solving the governing convection-dispersion transport equation. The model uses a mixed analytical and advanced numerical approach, and implements a transient partitioning of PAM mass between the canal water, the substrate soil, and potentially to open water bodies downstream of the application point. All source terms are modeled based on physical and chemical mechanisms as well as laboratory or field determined parameters. To more closely simulate field treatment of some canals, where PAM application moves upstream in time, the model is capable of implementing either a fixed or mobile upper boundary. In the latter treatment, the PAM can be added discretely or continuously in both time and space. A number of test situations have been simulated thus far, including theoretical and hypothetical cases for a wide range of conditions. The model also performed well when predicting PAM concentrations from a full-scale canal treatment experiment. The model provides a useful tool for predicting PAM fate and transport in water delivery canals, and therefore can play an important role in evaluating the efficacy of PAM application for water resources management

  8. Effect of cationic polyacrylamide on the processing and properties of nanocellulose films.

    PubMed

    Raj, Praveena; Varanasi, Swambabu; Batchelor, Warren; Garnier, Gil

    2015-06-01

    The use of high molecular weight cationic polyacrylamide (CPAM) was investigated to accelerate the drainage of nanocellulose (Microfibrillated Cellulose) suspensions into films. The mechanism was quantified and optimized by measuring the gel point, the lowest solids concentration at which a continuous network is formed. The flocculation of MFC was analysed as a function of the polyelectrolyte dosage, charge density and molecular weight as well as process parameters (drainage time) and material properties. The adsorption isotherms of CPAMs on nanocellulose and their zeta potential curves were also analysed as a function of CPAM charge and dosage. Measured CPAM adsorption capacities for the 50% and 10% charged 13MDa CPAM onto MFC were 5mg/g and 8mg/g, respectively, corresponding to adsorption coverage on cellulose of 0.14mg/m(2) and 0.22mg/m(2). The floc strength and drainability of MFC suspensions were quantified with the gel point as a function of CPAM properties. For all combinations of polyelectrolyte molecular weight and charge density, the gel point of a nanocellulose suspension goes through a minimum with increasing polymer dosage. The minimum gel point was independent of the polyelectrolyte charge density at constant molecular weight. However, it reduced with decreasing CPAM molecular weight, at a constant addition rate. The drainage time of a nanocellulose suspension into a film is reduced by 2/3 by halving the gel point from 0.2 to 0.1kg/m(3); this is due to the more flocculated suspension facilitating drainage between flocs. Nanocellulose films of increased porosity also result from reducing the gel point, signifying that the more open 3D structure of the flocculated cellulose suspension is retained upon drying the 2D film cellulose film structure.

  9. Nondenaturing agarose gel electrophoresis of RNA.

    PubMed

    Rio, Donald C; Ares, Manuel; Hannon, Gregory J; Nilsen, Timothy W

    2010-06-01

    INTRODUCTION Perhaps the most important and certainly the most often used technique in RNA analysis is gel electrophoresis. Because RNAs are negatively charged, they migrate toward the anode in the presence of electric current. The gel acts as a sieve to selectively impede the migration of the RNA in proportion to its mass, given that its mass is generally proportional to its charge. Because mass is approximately related to chain length, the length of an RNA is more generally determined by its migration. In addition, topology (i.e., circularity) can affect migration, making RNAs appear longer on the gel than they actually are. There are two common types of gel: polyacrylamide and agarose. For most applications involving RNAs of < or =600 nucleotides, denaturing acrylamide gels are most appropriate. In contrast, agarose gels are generally used to analyze RNAs of > or =600 nucleotides, and are especially useful for analysis of mRNAs (e.g., by Northern blotting). RNA analysis on agarose gels is essentially identical to DNA analysis (except that the gel boxes used must be dedicated to RNA work or to other ribonuclease-free work). Here we describe the use of straightforward Tris borate, EDTA (TBE) gels for routine analysis. These gels are appropriate for determining the quantity and integrity of RNA before using it for other applications. This procedure should not be used to determine size with accuracy, because the RNA will not remain in its extended state throughout the run.

  10. Superior hybrid hydrogels of polyacrylamide enhanced by bacterial cellulose nanofiber clusters.

    PubMed

    Yuan, Ningxiao; Xu, Lu; Zhang, Lu; Ye, Haowen; Zhao, Jianhao; Liu, Zhong; Rong, Jianhua

    2016-10-01

    Hybrid polyacrylamide/bacterial cellulose nanofiber clusters (PAM/BC) hydrogels with high strength, toughness and recoverability were synthesized by in situ polymerization of acrylamide monomer in BC nanofiber clusters suspension. The hybrid gels exhibited an extremely large elongation at break of 2200%, and a high fracture stress of 1.35MPa. Additionally, the original length of hydrogels could be recovered after releasing the tensile force. Compressive results showed that the PAM/BC hybrid gels could reach a strain of about 99% without break, and was able to completely recover its original shape immediately after releasing the compression force. The compressive stress at 99% reached as high as 30MPa. Nearly no hysteresis in cyclic compressive tests was observed with these hybrid gels. The FT-IR, XRD and TGA analysis showed that hydrogen bonds between the PAM chains and BC nanofiber clusters mainly contributed to the superior mechanical properties of hybrid hydrogels. The cell viability results suggested that PAM/BC hybrid hydrogel was benign for biomedical application. These PAM/BC hydrogels offer a great promise as biomaterials such as bone and cartilage repair materials.

  11. Antioxidant effect of green tea on polymer gel dosimeter

    NASA Astrophysics Data System (ADS)

    Samuel, E. J. J.; Sathiyaraj, P.; Deena, T.; Kumar, D. S.

    2015-01-01

    Extract from Green Tea (GTE) acts as an antioxidant in acrylamide based polymer gel dosimeter. In this work, PAGAT gel was used for investigation of antioxidant effect of GTE.PAGAT was called PAGTEG (Polyacrylamide green tea extract gel dosimeter) after adding GTE. Free radicals in water cause pre polymerization of polymer gel before irradiation. Polyphenols from GTE are highly effective to absorb the free radicals in water. THPC is used as an antioxidant in polymer gel dosimeter but here we were replaced it by GTE and investigated its effect by spectrophotometer. GTE added PAGAT samples response was lower compared to THPC added sample. To increase the sensitivity of the PAGTEG, sugar was added. This study confirmed that THPC was a good antioxidant for polymer gel dosimeter. However, GTE also can be used as an antioxidant in polymer gel if use less quantity (GTE) and add sugar as sensitivity enhancer.

  12. Multimaterial polyacrylamide: fabrication with electrohydrodynamic jet printing, applications, and modeling.

    PubMed

    Poellmann, Michael J; Johnson, Amy J Wagoner

    2014-09-01

    Micropatterned, multimaterial hydrogels have a wide range of applications, including the study of microenvironmental factors on cell behavior, and complex materials that rapidly change shape in response to fluid composition. This paper presents a method to fabricate microscale polyacrylamide features embedded in a second hydrogel of a different composition. An electrohydrodynamic jet (e-jet) printer was used to pattern hemispherical droplets of polyacrylamide prepolymer on a passive substrate. After photopolymerization, the droplets were backfilled with a second polyacrylamide mixture, the second mixture was polymerized and the sample was peeled off the substrate. Fluorescent and confocal microscopy confirmed multimaterial patterning, while scanning probe microscopy revealed a patterned topography with printed spots forming shallow wells. Finite element modeling was used to understand the mechanics of the formation of the topographical features during backfill and subsequent polymerization. Finally, polyacrylamide containing acrylic acid was used to demonstrate two applications of the micropatterned hydrogels: stimuli-responsive materials and patterned substrates for cell culture. The e-jet fabrication technique described here is a highly flexible, high resolution method for creating multimaterial hydrogels.

  13. Acrylamide monomer leaching from polyacrylamide-treated irrigation furrows

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Water-soluble polyacrylamide (WSPAM), used to reduce erosion in furrow irrigated fields and other agriculture applications, contain less than 0.05% Acrylamide monomer (AMD). The AMD, a potent neurotoxicant and suspected carcinogen, is readily dissolved and transported in flowing water. Deep percol...

  14. HRP-Mediated Synthesis of Starch-Polyacrylamide Graft Copolymers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Modified starch-based polymers can be engineered for specific properties by combining starch with synthetic polymers through graft copolymerization. Polyacrylamide grafted starch have received a great deal of applications in areas such as superabsorbent paper-making additives, drag reduction and te...

  15. HRP-Mediated Synthesis of Starch-Polyacrylamide Graft Copolymers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Modified starch-based polymers can be engineered for specific properties by combining starch with synthetic polymers through graft copolymerization. Polyacrylamide grafted starches have received a great deal of applications in areas such as superabsorbent paper-making additives, drag reduction and ...

  16. Graphitic carbon nitride embedded hydrogels for enhanced gel electrophoresis.

    PubMed

    Zarei, Mohammad; Ahmadzadeh, Hossein; Goharshadi, Elaheh K; Farzaneh, Ali

    2015-08-05

    Here, we show, for the first time, the use of graphitic carbon nitride (g-C3N4) nanosheets to improve the resolution and efficiency of protein separation in gel electrophoresis. By loading 0.04% (m/v) g-C3N4 nanosheets into the polyacrylamide gel at 25 °C, the thermal conductivity increased approximately 80% which resulted in 20% reduction in Joule heating and overall increase of separation efficiency. Also, polymerization of acrylamide occurred in the absence of tetramethylethylenediamine (TEMED) when the polyacrylamide gel contained g-C3N4 nanosheets. Hence, the g-C3N4 act simultaneously as a polymerization catalyst as well as heat sinks to lower Joule heating effect on band broadening.

  17. Sequencing of Proteins from Two-Dimensional Gels by Using in situ Digestion and Transfer of Peptides to Polyvinylidene Difluoride Membranes: Application to Proteins Associated with Sensitization in Aplysia

    NASA Astrophysics Data System (ADS)

    Kennedy, T. E.; Gawinowicz, M. A.; Barzilai, A.; Kandel, E. R.; Sweatt, J. D.

    1988-09-01

    We have developed a method for obtaining partial internal amino acid sequence data from proteins isolated directly from preparative two-dimensional polyacrylamide gels. Proteins from a crude cell homogenate are separated using preparative two-dimensional polyacrylamide gel electrophoresis. Then, the gel is stained with Coomassie blue and the protein spots of interest are cut out. The in situ protein is digested with Staphylococcus aureus V8 protease in a second polyacrylamide gel and the peptides are separated by one-dimensional polyacrylamide gel electrophoresis. The peptides are then electroblotted onto a polyvinylidene difluoride membrane, visualized using Coomassie blue, cut out, and sequenced using an automated gas phase sequencer. Using this method, we have obtained amino acid sequence data for two proteins that are altered after long-term sensitization: actin and Aplysia protein 407. In addition, we have obtained amino acid sequence data for rat protein 425, a protein that appears to be homologous to Aplysia protein 407.

  18. 21 CFR 872.3480 - Polyacrylamide polymer (modified cationic) denture adhesive.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Polyacrylamide polymer (modified cationic) denture... polymer (modified cationic) denture adhesive. (a) Identification. A polyacrylamide polymer (modified cationic) denture adhesive is a device composed of polyacrylamide polymer (modified cationic) intended...

  19. 21 CFR 872.3480 - Polyacrylamide polymer (modified cationic) denture adhesive.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Polyacrylamide polymer (modified cationic) denture... polymer (modified cationic) denture adhesive. (a) Identification. A polyacrylamide polymer (modified cationic) denture adhesive is a device composed of polyacrylamide polymer (modified cationic) intended...

  20. 21 CFR 872.3480 - Polyacrylamide polymer (modified cationic) denture adhesive.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Polyacrylamide polymer (modified cationic) denture... polymer (modified cationic) denture adhesive. (a) Identification. A polyacrylamide polymer (modified cationic) denture adhesive is a device composed of polyacrylamide polymer (modified cationic) intended...

  1. 21 CFR 872.3480 - Polyacrylamide polymer (modified cationic) denture adhesive.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Polyacrylamide polymer (modified cationic) denture... polymer (modified cationic) denture adhesive. (a) Identification. A polyacrylamide polymer (modified cationic) denture adhesive is a device composed of polyacrylamide polymer (modified cationic) intended...

  2. 21 CFR 872.3480 - Polyacrylamide polymer (modified cationic) denture adhesive.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Polyacrylamide polymer (modified cationic) denture... polymer (modified cationic) denture adhesive. (a) Identification. A polyacrylamide polymer (modified cationic) denture adhesive is a device composed of polyacrylamide polymer (modified cationic) intended...

  3. Feasibility of a DNA-Based Combinatorial Array Recognition Surface (CARS) in a Polyacrylamide Gel Matrix

    DTIC Science & Technology

    2007-12-12

    the phosphate backbone. wherever p.·J.Ttial hybrids naturally occur, via Taq DNA ligase (16). Ligation may not have been entirely necessary for the...CARS libraries, noncontiguous pieces can be ligaled togclher willi Taq DNA ligase . The lOp half o(lhe figure iIIu51Tates Ihe appearance o( a I-D CARS...cluding dideoxynucleotides. were from a "Si lver Sequence" kit purchased from Promega Corporation (Madison. WI). ThermliS aquaticus (Taq) DNA ligase was

  4. A platform to enable the pharmacological profiling of small molecules in gel-based electrophoretic mobility shift assays

    PubMed Central

    Foley, Timothy L.; Dorjsuren, Dorjbal; Dexheimer, Thomas; Burkart, Michael D.; Wight, William C.; Simeonov, Anton

    2017-01-01

    We describe a polyacrylamide gel casting cassette that overcomes limitations of commercially available gel electrophoresis equipment. This apparatus molds a single polyacrylamide gel that can evaluate more than 200 samples in parallel, is loaded with a multichannel pipettor, and is flexible with respect to composition of the separating matrix. We demonstrate its use to characterize inhibitors of enzymes that modify protein and nucleic acid substrates. Throughputs of greater than 1000 samples per day were achieved when this system was paired with a quantitative laser-based imaging system, yielding data of remarkable quality. PMID:27269812

  5. Gel-based versus gel-free proteomics: a review.

    PubMed

    Baggerman, Geert; Vierstraete, Evy; De Loof, Arnold; Schoofs, Liliane

    2005-12-01

    With the sequencing of the genome of over 150 organisms, the field of biology has been revolutionised. Instead of studying one gene or protein at the time, it is now possible to study the effect of physiological or pathological changes on the expression of all genes or proteins in the organism. Proteomics aims at the simultaneous analysis of all proteins expressed by a cell, tissue or organism in a specific physiological condition. Because proteins are the effector molecules in all organisms, it is evident that changes in the physiological condition of an organism will be reflected by changes in protein expression and/or processing. Since the formulation of the concept of proteomics in the mid 90's proteomics has relied heavily on 2 dimensional gel electrophoresis (2DGE) for the separation and visualization of proteins. 2DGE, however, has a number of inherent drawbacks. 2DGE is costly, fairly insensitive to low copy proteins and cannot be used for the entire proteome. Therefore, over the years, several gel-free proteomics techniques have been developed to either fill the gaps left by 2DGE or to entirely abolish the gel based techniques. This review summarizes the most important gel-free and gel-based proteomics techniques and compares their advantages and drawbacks.

  6. Sorption of copper(II) onto super-adsorbent of bentonite-polyacrylamide composites.

    PubMed

    Zhao, Guixia; Zhang, Hongxia; Fan, Qiaohui; Ren, Xuemei; Li, Jiaxing; Chen, Yixue; Wang, Xiangke

    2010-01-15

    In this work, bentonite embedded in the polyacrylamide (PAAm) gels was used as a novel adsorbent for the removal of Cu(II) from aqueous solution. The sorption and desorption of Cu(II) on bentonite-polyacrylamide (BENT-PAAm) was investigated as the function of pH, ionic strength, adsorbent content, Cu(II) concentrations and temperature. The results indicated that the sorption of Cu(II) on BENT-PAAm was strongly dependent on pH, ionic strength and temperature. The sorption increased from about 9% to 97% at pH ranging from 2.4 to 7. The sorption of Cu(II) on BENT-PAAm increased with increasing temperature and decreasing ionic strength. The sorption of Cu(II) on BENT and on BENT-PAAm was an endothermic and irreversible process. The results of desorption indicated that the adsorbed Cu(II) ions on solid particles were difficult to be desorbed from solid to liquid phase. From the comparison with BENT, BENT-PAAm showed higher sorption capacity with C(smax) increasing from 29 to 33 mg/g at pH 6.2 and from 11 to 20mg/g at pH 5.0 for the sorption of Cu(II) from BENT to BENT-PAAm composites. The average standard enthalpy change (Delta H degrees) and the entropy change (DeltaS degrees ) of Cu(II) sorption on BENT-PAAm are higher than those of Cu(II) sorption on BENT. The BENT-PAAm composites can be used as a super-adsorbent for the removal of Cu(II) from aqueous solution.

  7. Effect of gel structure of matrix orientation in pulsed alternating electric fields

    SciTech Connect

    Stellwagen, N.C.; Stellwagen, J.

    1993-12-31

    Four polymeric gels with different structures, LE agarose, HEEO agarose, beta-carrageenan, and polyacrylamide, were studied by transient electric birefringence to determine the importance of various structural features on the orientation of the gels in pulsed alternating electric fields. The birefrigence relaxation times observed for agarose gels in low voltage electric fields suggest that long fibers and/or domains, ranging up to tens of microns in size, are oriented by the electric field. The sign of the birefringence reverses when the direction of the electric field is reversed, suggesting that the oriented domains change their direction of orientation from parallel to perpendicular (or vice versa) when the polarity of the electric field is reversed. These anamalous orientation effects are observed with both types of agarose gels, but not with beta-carrageenan or polyacrylamide gels, suggesting that the alternating D,L galactose residues in the agarose backbone are responsible for the anomalies.

  8. Rapid agarose gel electrophoretic mobility shift assay for quantitating protein: RNA interactions.

    PubMed

    Ream, Jennifer A; Lewis, L Kevin; Lewis, Karen A

    2016-10-15

    Interactions between proteins and nucleic acids are frequently analyzed using electrophoretic mobility shift assays (EMSAs). This technique separates bound protein:nucleic acid complexes from free nucleic acids by electrophoresis, most commonly using polyacrylamide gels. The current study utilizes recent advances in agarose gel electrophoresis technology to develop a new EMSA protocol that is simpler and faster than traditional polyacrylamide methods. Agarose gels are normally run at low voltages (∼10 V/cm) to minimize heating and gel artifacts. In this study we demonstrate that EMSAs performed using agarose gels can be run at high voltages (≥20 V/cm) with 0.5 × TB (Tris-borate) buffer, allowing for short run times while simultaneously yielding high band resolution. Several parameters affecting band and image quality were optimized for the procedure, including gel thickness, agarose percentage, and applied voltage. Association of the siRNA-binding protein p19 with its target RNA was investigated using the new system. The agarose gel and conventional polyacrylamide gel methods generated similar apparent binding constants in side-by-side experiments. A particular advantage of the new approach described here is that the short run times (5-10 min) reduce opportunities for dissociation of bound complexes, an important concern in non-equilibrium nucleic acid binding experiments.

  9. The ARM Best Estimate 2-dimensional Gridded Surface

    SciTech Connect

    Xie,Shaocheng; Qi, Tang

    2015-06-15

    The ARM Best Estimate 2-dimensional Gridded Surface (ARMBE2DGRID) data set merges together key surface measurements at the Southern Great Plains (SGP) sites and interpolates the data to a regular 2D grid to facilitate data application. Data from the original site locations can be found in the ARM Best Estimate Station-based Surface (ARMBESTNS) data set.

  10. Optimal processing for gel electrophoresis images: Applying Monte Carlo Tree Search in GelApp.

    PubMed

    Nguyen, Phi-Vu; Ghezal, Ali; Hsueh, Ya-Chih; Boudier, Thomas; Gan, Samuel Ken-En; Lee, Hwee Kuan

    2016-08-01

    In biomedical research, gel band size estimation in electrophoresis analysis is a routine process. To facilitate and automate this process, numerous software have been released, notably the GelApp mobile app. However, the band detection accuracy is limited due to a band detection algorithm that cannot adapt to the variations in input images. To address this, we used the Monte Carlo Tree Search with Upper Confidence Bound (MCTS-UCB) method to efficiently search for optimal image processing pipelines for the band detection task, thereby improving the segmentation algorithm. Incorporating this into GelApp, we report a significant enhancement of gel band detection accuracy by 55.9 ± 2.0% for protein polyacrylamide gels, and 35.9 ± 2.5% for DNA SYBR green agarose gels. This implementation is a proof-of-concept in demonstrating MCTS-UCB as a strategy to optimize general image segmentation. The improved version of GelApp-GelApp 2.0-is freely available on both Google Play Store (for Android platform), and Apple App Store (for iOS platform).

  11. A new agarose matrix for single-strand conformation polymorphism (SSCP), heteroduplex (HTX), and gel shift analyses

    SciTech Connect

    Dumais, M.M.; White, H.W.; Rashid, M.R.

    1994-09-01

    Detection of mutation, by SSCP or heteroduplex analysis, is important in medical genetics and oncology. Analysis of DNA binding proteins is a powerful tool in molecular biology research. Traditionally, these methods are performed using nondenaturing gel electrophoresis on poly-acrylamide or polyacrylamide-type matrices. Here we report the development of a new agarose gel matrix that can be used for all three methods. SSCP analyses were performed using the prototype agarose gel matrix for wild-type, polymorphic, and mutant samples from c-Kras exon 12, p53 exons 8 and 9, and HOX2B. We performed SSCP analyses using both isotopic and nonisotopic methods. We also analyzed the samples by deliberate HTX formation and subsequent gel analysis. Using the prototype agarose matrix, we detected single and multiple DNA sequence variants in 150-350 bp fragments with an efficiency comparable to polyacrylamide gels run under similar conditions. For SSCP and HTX assays, we achieved optimal resolution in gels run in vertical formats. However, some HTX samples could be resolved in horizontal gel systems. In addition, based on our studies, we have developed a useful battery of controls and standards for quality control of SSCP and HTX assays. We analyzed several different DNA/protein complexes (SP1, AP2, and octamer binding protein) using the prototype agarose matrix. We obtained good resolution in both vertical and horizontal gel formats. The horizontal gel system is generally superior for this application, due to its ease of use and slightly better resolution. This new prototype gel matrix offers an alternative for researchers performing analyses that previously could only be done on polyacrylamide-type gel matrices. For some applications, this new matrix offers the ease of horizontal gel casting. For all applications, this matrix offers the safety of a nontoxic system and the reproducibility of a thermally gelling system.

  12. Cationic polyacrylamides enhance rates of starch and cellulose saccharification.

    PubMed

    Reye, John T; Maxwell, Kendra; Rao, Swati; Lu, Jian; Banerjee, Sujit

    2009-10-01

    Adding a cationic polyacrylamide (c-PAM) to either the amylase mediated hydrolysis of corn starch or the hydrolysis of wood fiber by cellulase can enhance the initial hydrolysis rates, although a rate decrease can occur under some conditions. Several c-PAMs can serve as catalysts and the same c-PAM can improve the efficiency of both amylase and cellulase. The initial amylase rate approximately doubles; the analogous cellulase hydrolysis rate increases by about 40%. c-PAMs increase the binding of enzyme to substrate.

  13. 4-dimensional spacetimes from 2-dimensional conformal null data

    NASA Astrophysics Data System (ADS)

    Goswami, Rituparno; Ellis, George F. R.

    2017-03-01

    In this paper we investigate whether the holographic principle proposed in string theory has a classical counterpart in general relativity theory. We show that there is a partial correspondence: at least in the case of vacuum Petrov type D spacetimes that admit a non-trivial Killing tensor, which encompass all the astrophysical black hole spacetimes, there exists a one-to-one correspondence between gravity in bulk and a 2-dimensional classical conformal scalar field on a null boundary.

  14. Exfoliation and Dispersion of 2-Dimensional Materials by Elevating Temperature

    NASA Astrophysics Data System (ADS)

    Kwon, Sanghyuk; Kim, Jinseon; Kwon, Hyukjoon; Lee, Changgu; Graphene Engneering Lab Team

    It is known that graphene and other 2-dimensional materials are hard to dissolve in water without using chemicals or surfactants. Here, we present a facile method to exfoliate and disperse those materials in water by simply controlling temperature. Graphene, when sonicated in water at high temperature (60°C), was edge-functionalized due to the extremely high temperature and pressure locally induced by ultrasonic cavitation, and dissolved in water stably even for longer than 1 month. However, it was not dispersed at low temperature(30°C) because of less cavitation and reduced sonochemical reaction. Other 2-dimensional materials, such as h-BN, MoS2, and other layered metal chalcogenides, were also well dissolved in water as graphene, but even at low temperature. Their stable solution is from the electric double layer because their relatively high insulating property. Also elevated storage temperature (60°C) improved the long-term dispersion stability compared to lower storage temperature (20°C) Exfoliation and Dispersion of 2-Dimensional Materials by Elevating Temperature.

  15. Microchannel gel electrophoretic separation systems and methods for preparing and using

    SciTech Connect

    Herr, Amy; Singh, Anup K; Throckmorton, Daniel J

    2013-09-03

    A micro-analytical platform for performing electrophoresis-based immunoassays was developed by integrating photopolymerized cross-linked polyacrylamide gels within a microfluidic device. The microfluidic immunoassays are performed by gel electrophoretic separation and quantifying analyte concentration based upon conventional polyacrylamide gel electrophoresis (PAGE). To retain biological activity of proteins and maintain intact immune complexes, native PAGE conditions were employed. Both direct (non-competitive) and competitive immunoassay formats are demonstrated in microchips for detecting toxins and biomarkers (cytokines, c-reactive protein) in bodily fluids (serum, saliva, oral fluids). Further, a description of gradient gels fabrication is included, in an effort to describe methods we have developed for further optimization of on-chip PAGE immunoassays. The described chip-based PAGE immunoassay method enables immunoassays that are fast (minutes) and require very small amounts of sample (less than a few microliters). Use of microfabricated chips as a platform enables integration, parallel assays, automation and development of portable devices.

  16. Microchannel gel electrophoretic separation systems and methods for preparing and using

    SciTech Connect

    Herr, Amy E; Singh, Anup K; Throckmorton, Daniel J

    2015-02-24

    A micro-analytical platform for performing electrophoresis-based immunoassays was developed by integrating photopolymerized cross-linked polyacrylamide gels within a microfluidic device. The microfluidic immunoassays are performed by gel electrophoretic separation and quantifying analyte concentration based upon conventional polyacrylamide gel electrophoresis (PAGE). To retain biological activity of proteins and maintain intact immune complexes, native PAGE conditions were employed. Both direct (non-competitive) and competitive immunoassay formats are demonstrated in microchips for detecting toxins and biomarkers (cytokines, c-reactive protein) in bodily fluids (serum, saliva, oral fluids). Further, a description of gradient gels fabrication is included, in an effort to describe methods we have developed for further optimization of on-chip PAGE immunoassays. The described chip-based PAGE immunoassay method enables immunoassays that are fast (minutes) and require very small amounts of sample (less than a few microliters). Use of microfabricated chips as a platform enables integration, parallel assays, automation and development of portable devices.

  17. Environmental degradation of polyacrylamides. 1. Effects of artificial environmental conditions: temperature, light, and pH.

    PubMed

    Smith, E A; Prues, S L; Oehme, F W

    1996-11-01

    A polyacrylamide thickening agent (PATA) was formulated at four concentrations in distilled-deionized water, without and with a glyphosate-surfactant herbicide (GH). Over a 6-week period, these mixtures were exposed to various controlled temperature and light conditions. Acrylamide concentration, ammonium concentration, and pH were measured at weekly intervals to assess the degradation of polyacrylamide and acrylamide. Satellite studies were conducted to examine the effect of altered pH on solutions of PATA (i.e., does pH promote polyacrylamide depolymerization?) and GH binding to amine groups (i.e., protection from degradation). The results of these studies suggest that polyacrylamide can degrade to acrylamide by thermal and photolytic effects, that changes in pH do not promote the depolymerization of polyacrylamide, and that GH does protect polyacrylamide and acrylamide from environmental degradation. Statistically there was no linear correlation between the various parameters measured.

  18. Increase in local protein concentration by field-inversion gel electrophoresis.

    PubMed

    Tsai, Henghang; Leung, Hon-Chiu Eastwood

    2012-01-01

    Proteins that migrate through cross-linked polyacrylamide gels (PAGs) under the influence of a constant electric field experience negative factors, such as diffusion and nonspecific trapping in the gel matrix. These negative factors reduce protein concentrations within a defined gel volume with increasing migration distance and, therefore, decrease protein recovery efficiency. Here, we describe the enhancement of protein separation efficiency up to twofold in conventional one-dimensional PAG electrophoresis (1D PAGE), two-dimensional (2D) PAGE, and native PAGE by implementing pulses of inverted electric field during gel electrophoresis.

  19. The latest advancements in proteomic two-dimensional gel electrophoresis analysis applied to biological samples.

    PubMed

    Santucci, Laura; Bruschi, Maurizio; Ghiggeri, Gian Marco; Candiano, Giovanni

    2015-01-01

    Two-dimensional gel electrophoresis (2DE) is one of the fundamental approaches in proteomics for the separation and visualization of complex protein mixtures. Proteins can be analyzed by 2DE using isoelectric focusing (IEF) in the first dimension, combined to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the second dimension, gel staining (silver and Coomassie), image analysis, and 2DE gel database. High-resolution 2DE can resolve up to 5,000 different proteins simultaneously (∼2,000 proteins routinely), and detect and quantify <1 ng of protein per spot. Here, we describe the latest developments for a more complete analysis of biological fluids.

  20. Effects of polymer concentration on the morphology of calcium phosphate crystals formed in polyacrylamide hydrogels

    NASA Astrophysics Data System (ADS)

    Yokoi, Taishi; Kawashita, Masakazu; Ohtsuki, Chikara

    2013-11-01

    Growing crystals in hydrogels is an attractive method to form inorganic solids with designed morphology under ambient conditions. Precipitation of the inorganic solids in a hydrogel matrix can be regarded as mimicking the process of biomineralization. In the construction of biominerals, an organic template composed of insoluble macromolecules is used to control the crystal growth of the inorganic compounds. The morphological control in biomineralization can be applied to artificial reaction systems. In this study, the morphology of calcium phosphate crystals formed in polymeric hydrogels of various polymer concentrations was investigated. Spherical octacalcium phosphate (OCP) precipitated in the polyacrylamide (PAAm) hydrogels. Fibrous crystals gradually covered the surface of the spherical crystals as the polymer concentration of the gel increased. The morphology of the OCP crystals changed from sea urchin shapes to wool-ball shapes with increasing PAAm concentration. The morphological change is generated by the template effect of the polymer wall, which is made up of stacked PAAm sheets, surrounding the spherical OCP crystals.

  1. Isolation and Characterization of Polyacrylamide-Degrading Bacteria from Dewatered Sludge

    PubMed Central

    Yu, Feng; Fu, Ruimin; Xie, Yun; Chen, Wuling

    2015-01-01

    Polyacrylamide (PAM) is a water-soluble polymer that is widely used as a flocculant in sewage treatment. The accumulation of PAM affects the formation of dewatered sludge and potentially produces hazardous monomers. In the present study, the bacterial strain HI47 was isolated from dewatered sludge. This strain could metabolize PAM as its sole nutrient source and was subsequently identified as Pseudomonas putida. The efficiency of PAM degradation was 31.1% in 7 days and exceeded 45% under optimum culture condition (pH 7.2, 39 °C and 100 rpm). The addition of yeast extract and glucose improved the bacterial growth and PAM degradation. The degraded PAM samples were analyzed by gel-filtration chromatography, Fourier transform infrared and high-performance liquid chromatography. The results showed that high-molecular-weight PAM was partly cleaved to small molecular oligomer derivatives and part of the amide groups of PAM had been converted to carboxyl groups. The biodegradation did not accumulate acrylamide monomers. Based on the SDS-PAGE and N-terminal sequencing results, the PAM amide groups were converted into carboxyl groups by a PAM-induced extracellular enzyme from the aliphatic amidase family. PMID:25893998

  2. Aerosol gels

    NASA Technical Reports Server (NTRS)

    Sorensen, Christopher M. (Inventor); Chakrabarti, Amitabha (Inventor); Dhaubhadel, Rajan (Inventor); Gerving, Corey (Inventor)

    2010-01-01

    An improved process for the production of ultralow density, high specific surface area gel products is provided which comprises providing, in an enclosed chamber, a mixture made up of small particles of material suspended in gas; the particles are then caused to aggregate in the chamber to form ramified fractal aggregate gels. The particles should have a radius (a) of up to about 50 nm and the aerosol should have a volume fraction (f.sub.v) of at least 10.sup.-4. In preferred practice, the mixture is created by a spark-induced explosion of a precursor material (e.g., a hydrocarbon) and oxygen within the chamber. New compositions of matter are disclosed having densities below 3.0 mg/cc.

  3. Analysis of soybean embryonic axis proteins by two-dimensional gel electrophoresis and mass spectrometry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A proteomic approach based on two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) for protein separation and subsequent mass spectrometry (MS) for protein identification was applied to establish a proteomic reference map for the soybean embryonic axis. Proteins were extracted from dissecte...

  4. Influence of graphene-oxide nanosheets impregnation on properties of sterculia gum-polyacrylamide hydrogel formed by radiation induced polymerization.

    PubMed

    Singh, Baljit; Singh, Baldev

    2017-06-01

    Present work is an attempt, to explore the potential of graphene oxide nanoplates impregnation, on the mechanical and drug delivery properties of sterculia gum-polyacrylamide composite hydrogel formed by radiation induced polymerization. These polymers were characterized by SEM, cryo-SEM, AFM, FTIR's, (13)C NMR and swelling studies. Release profile of an anticancer drug 'gemcitabine' was studied to determine the drug release mechanism and best fit kinetic model. Furthermore, some important biomedical properties of the polymers such as blood compatibility, mucoadhesion, antioxidant properties and gel strength were also studied. Impregnation of GO into sterculia gum-poly(AAm) hydrogels decreased the swelling of hydrogels but improved the mechanical, drug loading and drug release properties of the hydrogels. Release of gemcitabine from drug loaded hydrogels occurred through non-Fickian diffusion mechanism and release profile was best fitted in first order kinetic model. These hydrogels have been found as haemocompatible, mucoadhesive, and antioxidant in nature.

  5. Environmental degradation of polyacrylamides. II. Effects of environmental (outdoor) exposure.

    PubMed

    Smith, E A; Prues, S L; Oehme, F W

    1997-06-01

    The environmental fate of a polyacrylamide thickening agent (PATA), formulated without and with a glyphosate-surfactant herbicide (GH), was examined under various environmental situations: formulation in surface water and ground water, volatility, and soil mobility. Environmental Fate of PATA in Surface Water and Ground Water: PATA was formulated at four concentrations in distilled-deionized water, three surface water samples, and two ground water samples, without and with a GH. Solutions were placed in glass bottles, covered with plastic wrap, and exposed to environmental (outdoor) conditions for 6 weeks. Acrylamide and ammonium concentration, pH, and bacterial and fungal populations were measured weekly. All solutions in this portion of the study had a homogeneous milky appearance but the conclusions of the study were nearly transparent. The results of this study suggest that polyacrylamide can degrade to acrylamide under environmental conditions. Statistically, there was no linear correlation between the various parameters measured. Volatility: PATA was formulated without and with GH. Each solution plus an acrylamide standard (positive control) was placed in a glass beaker and exposed to environmental (outdoor) conditions for 6 days. Acrylamide concentration, ammonium concentration, pH, and solution volume were measured daily. Acrylamide and ammonium concentrations increased during the study in all formulations, except when solutions evaporated to dryness. pH did not change greatly over the course of the study for these samples. Those solutions containing PATA had a homogeneous milky appearance but by the conclusions of the study were nearly transparent. This suggests a physical structural change in the polymer. Soil Mobility: PATA formulated with GH was also applied to soil columns and soil boxes containing sand, Eudora sandy loam, Eudora sandy clay, and Kohola silt loam. Acrylamide could be detected by Day 2 in all soil columns. Acrylamide could not be

  6. Recycling of superfine resolution agarose gel.

    PubMed

    Seng, T-Y; Singh, R; Faridah, Q Z; Tan, S-G; Alwee, S S R S

    2013-07-08

    Genetic markers are now routinely used in a wide range of applications, from forensic DNA analysis to marker-assisted plant and animal breeding. The usual practice in such work is to extract the DNA, prime the markers of interest, and sift them out by electrically driving them through an appropriate matrix, usually a gel. The gels, made from polyacrylamide or agarose, are of high cost, limiting their greater applications in molecular marker work, especially in developing countries where such technology has great potential. Trials using superfine resolution (SFR) agarose for SSR marker screening showed that it is capable of resolving SSR loci and can be reused up to 14 times, thus greatly reducing the cost of each gel run. Furthermore, for certain applications, low concentrations of agarose sufficed and switching to lithium borate buffer, instead of the conventional Tris-borate-ethylenediaminetetraacetic acid buffer, will further save time and cost. The 2.5% gel was prepared following the Agarose SFR(TM) manual by adding 2.5 g agarose powder into 100 mL 1X lithium borate buffer in a 250-mL flask with rapid stirring. Two midigels (105 x 83 mm, 17 wells) or 4 minigels (50 x 83 mm, 8 wells), 4 mm thickness can be prepared from 100 mL gel solution. A total of 1680 PCR products amplified using 140 SSR markers from oil palm DNA samples were tested in this study using SFR recycled gel. As average, the gel can be recycled 8 times with good resolution, but can be recycled up to 14 times before the resolutions get blurred.

  7. Low-velocity super-lubrication of sodium-alginate/polyacrylamide ionic-covalent hybrid double-network hydrogels.

    PubMed

    Li, Xuefeng; Wu, Chu; Yang, Qian; Long, Shijun; Wu, Chonggang

    2015-04-21

    Structural and frictional behaviours of sodium alginate (SA)/polyacrylamide (PAAm) ionic-covalent hybrid, sequential double-network (DN) hydrogels against glass have been investigated in water, NaCl and CaCl2 aqueous solutions using a rotational rheometer. Dilution of adsorptive elastohydrodynamic friction for the PAAm covalent network with repulsive hydrodynamic lubrication for the minor SA ionic network was found to control the frictional stresses of the SA/PAAm gels within between those of the SA and PAAm single-network gels. A tentative qualitative model was proposed to describe the impact of ionic environmental solution on the frictional behaviour of the hybrid gel by selectively affecting the SA-network structure and friction. It was revealed that strong Debye shielding in the NaCl solution significantly reduced the thickness of the electric double layer for hydrodynamic lubrication of the SA network, which made the SA/PAAm gel's friction the highest among the three solutions. Dramatically increased ionic cross-linking of the SA network in the CaCl2 solution, although effectively mediated by the PAAm-network flexible skeleton, still functioned partially to conserve a portion of the SA fractional boundary-friction at the interface, making the friction of the hybrid gel intermediate among the three solutions. In contrast, extreme hydration of the SA network in water sharply increased the volume fraction of its unshielded hydrodynamic lubrication at the interface, which greatly reduced the SA/PAAm's friction to the lowest among the three solutions. We have thus incorporated for the first time both super-lubrication (frictional coefficients of below 10(-2) over low sliding-velocities of 3 × 10(-5) to 2 × 10(-3) m s(-1)) and previously reported high fracture energy (over 9000 J m(-2)) into a single ionic-covalent hybrid DN hydrogel, which is the SA/PAAm (∼1/8.5 w/w) gel in water. Effects of inversion of DN-formation sequence further indicated that frictional

  8. Gadolinium-loaded gel scintillators for neutron and antineutrino detection

    SciTech Connect

    Riddle, Catherine Lynn; Akers, Douglas William; Demmer, Ricky Lynn; Paviet, Patricia Denise; Drigert, Mark William

    2016-11-29

    A gadolinium (Gd) loaded scintillation gel (Gd-ScintGel) compound allows for neutron and gamma-ray detection. The unique gel scintillator encompasses some of the best features of both liquid and solid scintillators, yet without many of the disadvantages associated therewith. Preferably, the gel scintillator is a water soluble Gd-DTPA compound and water soluble fluorophores such as: CdSe/ZnS (or ZnS) quantum dot (Q-dot) nanoparticles, coumarin derivatives 7-hydroxy-4-methylcoumarin, 7-hydroxy-4-methylcoumarin-3-acetic acid, 7-hydroxycoumarin-3-carboxylic acid, and Alexa Fluor 350 as well as a carbostyril compound, carbostyril 124 in a stable water-based gel, such as methylcellulose or polyacrylamide polymers. The Gd-loaded ScintGel allows for a homogenious distribution of the Gd-DTPA and the fluorophores, and yields clean fluorescent emission peaks. A moderator, such as deuterium or a water-based clear polymer, can be incorporated in the Gd-ScintGel. The gel scintillators can be used in compact detectors, including neutron and antineutrino detectors.

  9. Regioselective immobilization of short oligonucleotides to acrylic copolymer gels.

    PubMed Central

    Timofeev, E; Kochetkova, S V; Mirzabekov, A D; Florentiev, V L

    1996-01-01

    Four types of polyacrylamide or polydimethyl-acrylamide gels for regioselective (by immobilization at the 3' end) of short oligonucleotides have been designed for use in manufacturing oligonucleotide microchips. Two of these supports contain amino or aldehyde groups in the gel, allowing coupling with oligonucleotides bearing aldehyde or amino groups, respectively, in the presence of a reducing agent. The aldehyde gel support showed a higher immobilization efficiency relative to the amino gel. Of all reducing agents tested, the best results were obtained with a pyridine-borane complex. The other supports are based on an acrylamide gel activated with glutaraldehyde or a hydroxyalkyl-functionalized gel treated with mesyl chloride. The use of dimethylacrylamide instead of acrylamide allows subsequent gel modifications in organic solvents. All the immobilization methods are easy and simple to perform, give high and reproducible yields, allow long durations of storage of the activated support, and provide high stability of attachment and low non-specific binding. Although these gel supports have been developed for preparing oligonucleotide microchips, they may be used for other purposes as well. PMID:8774893

  10. A novel mechanism of “metal gel-shift” by histidine-rich Ni2+-binding Hpn protein from Helicobacter pylori strain SS1

    PubMed Central

    Ito, Yuki; Masumoto, Junya; Morita, Eugene Hayato; Hayashi, Hidenori

    2017-01-01

    Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) is a universally used method for determining approximate molecular weight (MW) in protein research. Migration of protein that does not correlate with formula MW, termed “gel shifting” appears to be common for histidine-rich proteins but not yet studied in detail. We investigated “gel shifting” in Ni2+-binding histidine-rich Hpn protein cloned from Helicobacter pylori strain SS1. Our data demonstrate two important factors determining “gel shifting” of Hpn, polyacrylamide-gel concentration and metal binding. Higher polyacrylamide-gel concentrations resulted in faster Hpn migration. Irrespective of polyacrylamide-gel concentration, preserved Hpn-Ni2+ complex migrated faster (3–4 kDa) than apo-Hpn, phenomenon termed “metal gel-shift” demonstrating an intimate link between Ni2+ binding and “gel shifting”. To examine this discrepancy, eluted samples from corresponding spots on SDS-gel were analyzed by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS). The MW of all samples was the same (6945.66±0.34 Da) and identical to formula MW with or without added mass of Ni2+. MALDI-TOF-MS of Ni2+-treated Hpn revealed that monomer bound up to six Ni2+ ions non-cooperatively, and equilibrium between protein-metal species was reliant on Ni2+ availability. This corroborates with gradually increased heterogeneity of apo-Hpn band followed by compact "metal-gel shift" band on SDS-PAGE. In view of presented data metal-binding and “metal-gel shift” models are discussed. PMID:28207866

  11. Embedded ceria nanoparticles in gel improve electrophoretic separation: a preliminary demonstration.

    PubMed

    Zarei, Mohammad; Ahmadzadeh, Hossein; Goharshadi, Elaheh K

    2015-07-07

    Slab gel electrophoresis is still the gold standard method for the separation of biomolecules such as proteins and DNA with advantages such as simplicity, affordability, and high throughput, but it suffers from inadequate separation speed and resolution. Single capillary gel electrophoresis, on the other hand, offers faster separation time and improved resolution at the expense of higher cost and loss of high throughput capability. The high surface to volume ratio of the capillary causes improved heat dissipation leading to a reduced Joule heating and a higher resolution. Here, for the first time, we show the use of dispersed ceria nanoparticles (NPs) to improve the resolution and speed of protein separation in slab gel electrophoresis. We measured the rheological parameters of separation medium in order to find a meaningful relationship between viscosity changes, Joule heating, and band broadening. The results showed that ceria NPs decrease the viscosity of polyacrylamide gel. By loading 0.03% (w/v) ceria NPs into polyacrylamide gel at 25 °C, the viscosity decreased 22% and the thermal conductivity increased to 81%, which resulted in 35% reduction in Joule heating and 47% increase in resolution. This work is a cross disciplinary of theoretical physical chemistry for thermal conductivity and rheological measurements of PA and ceria suspensions and application in slab gel electrophoresis. We report here, for the first time, that embedded NPs in PA gel could potentially interface high throughput capability of slab gel electrophoresis with high separation speed of single capillary electrophoresis.

  12. Centrifugal methods and devices for rapid in-gel digestion of proteins.

    PubMed

    Lazarev, Alexander V; Rejtar, Tomas; Dai, Shujia; Karger, Barry L

    2009-03-01

    Modern proteomic research frequently relies upon separation of proteins in a polyacrylamide gel matrix followed by in-gel enzymatic digestion and extraction of peptides for subsequent analysis by MS. In this work, we propose a novel semi-automated method of mechanical processing of gel bands by passing these bands through a specially designed centrifugal device termed a Gel Shredder prior to digestion and extraction of peptides. Such a device allows integrated washing, destaining and shredding of gel bands into uniform blocks of controlled size, approximately 150-300 microm, prior to the enzymatic digestion and extraction of peptides. Shredding into uniform blocks increases the surface area of the gel pieces and promotes improved gel rehydration, allowing improved diffusion of the proteolytic enzymes and solvent into the gel lattice. We demonstrate that the new method substantially reduces the time spent on tedious manual handling of gel bands, while minimizing the risk of sample contamination. The performance of the Gel Shredder has been compared with a conventional in-gel digestion protocol using several standard proteins and a complex proteomic sample in terms of relative quantitation by either MALDI-TOF/TOF or nanoLC-ESI IT-Fourier transformation ion cyclotron resonance MS. It is shown that significant time savings and improved peptide recovery can be obtained for many proteins using the Gel Shredder compared with the traditional in-gel digestion protocol.

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

    SciTech Connect

    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.

  14. Polyacrylamide medium for the electrophoretic separation of biomolecules

    DOEpatents

    Madabhushi, Ramakrishna S.; Gammon, Stuart A.

    2003-11-11

    A polyacryalmide medium for the electrophoretic separation of biomolecules. The polyacryalmide medium comprises high molecular weight polyacrylamides (PAAm) having a viscosity average molecular weight (M.sub.v) of about 675-725 kDa were synthesized by conventional red-ox polymerization technique. Using this separation medium, capillary electrophoresis of BigDye DNA sequencing standard was performed. A single base resolution of .about.725 bases was achieved in .about.60 minute in a non-covalently coated capillary of 50 .mu.m i.d., 40 cm effective length, and a filed of 160 V/cm at 40.degree. C. The resolution achieved with this formulation to separate DNA under identical conditions is much superior (725 bases vs. 625 bases) and faster (60 min. vs. 75 min.) to the commercially available PAAm, such as supplied by Amersham. The formulation method employed here to synthesize PAAm is straight-forward, simple and does not require cumbersome methods such as emulsion polymerizaiton in order to achieve very high molecular weights. Also, the formulation here does not require separation of PAAm from the reaction mixture prior to reconstituting the polymer to a final concentration. Furthermore, the formulation here is prepared from a single average mol. wt. PAAm as opposed to the mixture of two different average mo. wt. PAAm previously required to achieve high resolution.

  15. Antimicrobial activity of silver/starch/polyacrylamide nanocomposite.

    PubMed

    Abdel-Halim, E S; Al-Deyab, Salem S

    2014-07-01

    A novel silver/starch/polyacrylamide nanocomposite hydrogel was prepared by grafting acrylamide onto starch in presence of silver nitrate by use of ammonium persulphate as an initiator and N,N-methylene-bisacrylamide as a crosslinking agent, then reducing the silver ions enclosed in the hydrogel structure to silver nanoparticles by treating the hydrogel with sodium hydroxide solution. All factors which affect the grafting/crosslinking reaction were optimized and the concentration of silver ion was changed from 0ppm to 50ppm. The produced nanocomposite hydrogel was characterized for its nanosilver content and the UV-spectra showed similar absorption spectra at wavelength 405nm for all AgNO3 concentrations but the plasmon showed increase in the intensity of the absorption peak as AgNO3 concentration incorporated to the hydrogel structure increases. The nanocomposite hydrogel was also characterized for its antimicrobial activity toward two types of bacteria and two types of fungi. The results showed that the hydrogel with 0ppm silver content has no antimicrobial activity, and that the antimicrobial activity expressed as inhibition zone increases as the silver content increases from 5ppm to 50ppm.

  16. A new staining and evaluating procedure for protein gel electropherograms based on the pyrogallol red-molybdate complex.

    PubMed

    Csiba, A; Szécsényi-Nagy, L

    1989-01-01

    A new method is reported for staining and evaluating gel electropherograms of proteins. With pyrogallol red-molybdate reagent the gel-embedded proteins are transformed into a derivative of blue colour. After destaining, the blue-coloured proteins are well visible against a colourless background and can be quantified by densitometry with high reliability. The quantity of the coloured protein is directly proportional to the height of peaks in the densitogram. Colour intensity is concentration dependent. The measurement range of serum albumin was 1 to 50 micrograms/tube and 10 to 100 micrograms/slab in polyacrylamide gel disc electrophoresis and agar gel electrophoresis, respectively.

  17. Fully Automated Portable Comprehensive 2-Dimensional Gas Chromatography Device.

    PubMed

    Lee, Jiwon; Zhou, Menglian; Zhu, Hongbo; Nidetz, Robert; Kurabayashi, Katsuo; Fan, Xudong

    2016-10-06

    We developed a fully automated portable 2-dimensional (2-D) gas chromatography (GC x GC) device, which had a dimension of 60 cm × 50 cm × 10 cm and weight less than 5 kg. The device incorporated a micropreconcentrator/injector, commercial columns, micro-Deans switches, microthermal injectors, microphotoionization detectors, data acquisition cards, and power supplies, as well as computer control and user interface. It employed multiple channels (4 channels) in the second dimension ((2)D) to increase the (2)D separation time (up to 32 s) and hence (2)D peak capacity. In addition, a nondestructive flow-through vapor detector was installed at the end of the (1)D column to monitor the eluent from (1)D and assist in reconstructing (1)D elution peaks. With the information obtained jointly from the (1)D and (2)D detectors, (1)D elution peaks could be reconstructed with significantly improved (1)D resolution. In this Article, we first discuss the details of the system operating principle and the algorithm to reconstruct (1)D elution peaks, followed by the description and characterization of each component. Finally, 2-D separation of 50 analytes, including alkane (C6-C12), alkene, alcohol, aldehyde, ketone, cycloalkane, and aromatic hydrocarbon, in 14 min is demonstrated, showing the peak capacity of 430-530 and the peak capacity production of 40-80/min.

  18. Waste-Activated Sludge Fermentation for Polyacrylamide Biodegradation Improved by Anaerobic Hydrolysis and Key Microorganisms Involved in Biological Polyacrylamide Removal

    PubMed Central

    Dai, Xiaohu; Luo, Fan; Zhang, Dong; Dai, Lingling; Chen, Yinguang; Dong, Bin

    2015-01-01

    During the anaerobic digestion of dewatered sludge, polyacrylamide (PAM), a chemical conditioner, can usually be consumed as a carbon and nitrogen source along with other organic matter (e.g., proteins and carbohydrates in the sludge). However, a significant accumulation of acrylamide monomers (AMs) was observed during the PAM biodegradation process. To improve the anaerobic hydrolysis of PAM, especially the amide hydrolysis process, and to avoid the generation of the intermediate product AM, a new strategy is reported herein that uses an initial pH of 9, 200 mg COD/L of PAM and a fermentation time of 17 d. First, response surface methodology (RSM) was applied to optimize PAM removal in the anaerobic digestion of the sludge. The biological hydrolysis of PAM reached 86.64% under the optimal conditions obtained from the RSM. Then, the mechanisms for the optimized parameters that significantly improved the biological hydrolysis of PAM were investigated by the synergistic effect of the main organic compounds in the sludge, the floc size distribution, and the enzymatic activities. Finally, semi-continuous-flow experiments for a microbial community study were investigated based on the determination of key microorganisms involved in the biological hydrolysis of PAM. PMID:26144551

  19. Two-dimensional gel electrophoresis: vertical isoelectric focusing.

    PubMed

    Dorri, Yaser

    2012-01-01

    Two-dimensional gel electrophoresis (2-DE) is one of the most powerful tools for separating proteins based on their size and charge. 2-DE is very useful to separate two proteins with identical molecular weights but different charges, which cannot be achieved with just sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Here, a simpler and easier version of 2-DE is presented which is also faster than all the currently available techniques. In this modified version of 2-DE, isoelectric focusing is carried out in the first dimension using a vertical SDS-PAGE apparatus. Following the first-dimensional IEF, each individual lane is excised from the IEF gel and, after a 90° rotation, is inserted into a second-dimensional SDS-PAGE, which can be stained with Coomassie Brilliant Blue for protein analysis or immunoblotted for further analysis. This version of IEF can be run in less than 2 h compared to the overnight run required by O'Farrell's method. Difficult tube gel casting and gel extrusion as well as tube gel distortion are eliminated in our method. This method is simpler, faster, and inexpensive. Both dimensions can be done on the same SDS-PAGE apparatus, and up to ten samples can be run simultaneously using one gel.

  20. Bioactive polyacrylamide hydrogels with gradients in mechanical stiffness.

    PubMed

    Diederich, Vincent E G; Studer, Peter; Kern, Anita; Lattuada, Marco; Storti, Giuseppe; Sharma, Ram I; Snedeker, Jess G; Morbidelli, Massimo

    2013-05-01

    We propose a novel, single step method for the production of polyacrylamide hydrogels with a gradient in mechanical properties. In contrast to already existing techniques such as UV photo-polymerization with photomasks (limited penetration depth) or microfluidic gradient mixers (complex microfluidic chip), this technique is not suffering such limitations. Young's modulus of the hydrogels was varied by changing the total monomer concentration of the hydrogel precursor solution. Using programmable syringe pumps, the total monomer concentration in the solution fed to the hydrogel mold was varied from 16 wt% down to 5 wt% over the feeding time to obtain a gradient in compliance ranging from 150 kPa down to 20 kPa over a length of 10 mm down to 2.5 mm. Polymerization was achieved with the dual initiation system composed of ammonium persulfate and N,N,N',N'-tetramethylethylenediamine, which were both fed through separate capillaries to avoid premature polymerization. Functionalized with the model ligand collagen I, the substrates were bioactive and supported the attachment of human foreskin fibroblasts (around 30% of the cells seeded attached after 1 h). A kinetic morphology study on homogeneous hydrogels of different stiffness's indicated that fibroblasts tend to spread to their final size within 2 h on stiff substrates, while the spreading time was much longer (ca. 4-5 h) on soft substrates. These trends were confirmed on hydrogels with compliance gradients, showing well spread fibroblasts on the stiff end of the hydrogel after 2 h, while the cells on the soft end still had small area and rounded morphology.

  1. Turbidimetric determination of anionic polyacrylamide in low carbon soil extracts.

    PubMed

    Kang, Jihoon; Sowers, Tyler D; Duckworth, Owen W; Amoozegar, Aziz; Heitman, Joshua L; McLaughlin, Richard A

    2013-11-01

    Concerns over runoff water quality from agricultural lands and construction sites have led to the development of improved erosion control practices, including application of polyacrylamide (PAM). We developed a quick and reliable method for quantifying PAM in soil extracts at low carbon content by using a turbidimetric reagent, Hyamine 1622. Three high-molecular weight anionic PAMs differing in charge density (7, 20, and 50 mol%) and five water matrices, deionized (DI) water and extracts from four different soils, were used to construct PAM calibration curves by reacting PAM solutions with hyamine and measuring turbidity development from the PAM-hyamine complex. The PAM calibration curve with DI water showed a strong linear relationship ( = 0.99), and the sensitivity (slope) of calibration curves increased with increasing PAM charge density with a detection limit of 0.4 to 0.9 mg L. Identical tests with soil extracts showed the sensitivity of the hyamine method was dependent on the properties of the soil extract, primarily organic carbon concentration. Although the method was effective in mineral soils, the highest charge density PAM yielded a more reliable linear relationship ( > 0.97) and lowest detection limit (0.3 to 1.2 mg L), compared with those of the lower charge density PAMs (0.7 to 23 mg L). Our results suggest that the hyamine test could be an efficient method for quantifying PAM in environmental soil water samples as long as the organic carbon in the sample is low, such as in subsurface soil material often exposed at construction sites.

  2. Probing structure-antifouling activity relationships of polyacrylamides and polyacrylates.

    PubMed

    Zhao, Chao; Zhao, Jun; Li, Xiaosi; Wu, Jiang; Chen, Shenfu; Chen, Qiang; Wang, Qiuming; Gong, Xiong; Li, Lingyan; Zheng, Jie

    2013-07-01

    We have synthesized two different polyacrylamide polymers with amide groups (polySBAA and polyHEAA) and two corresponding polyacrylate polymers without amide groups (polySBMA and polyHEA), with particular attention to the evaluation of the effect of amide group on the hydration and antifouling ability of these systems using both computational and experimental approaches. The influence of polymer architectures of brushes, hydrogels, and nanogels, prepared by different polymerization methods, on antifouling performance is also studied. SPR and ELISA data reveal that all polymers exhibit excellent antifouling ability to repel proteins from undiluted human blood serum/plasma, and such antifouling ability can be further enhanced by presenting amide groups in polySBAA and polyHEAA as compared to polySBMA and polyHEA. The antifouling performance is positively correlated with the hydration properties. Simulations confirm that four polymers indeed have different hydration characteristics, while all presenting a strong hydration overall. Integration of amide group with pendant hydroxyl or sulfobetaine group in polymer backbones is found to increase their surface hydration of polymer chains and thus to improve their antifouling ability. Importantly, we present a proof-of-concept experiment to synthesize polySBAA nanogels, which show a switchable property between antifouling and pH-responsive functions driven by acid-base conditions, while still maintaining high stability in undiluted fetal bovine serum and minimal toxicity to cultured cells. This work provides important structural insights into how very subtle structural changes in polymers can yield great improvement in biological activity, specifically the inclusion of amide group in polymer backbone/sidechain enables to obtain antifouling materials with better performance for biomedical applications.

  3. Check dam and polyacrylamide performance under simulated stormwater runoff.

    PubMed

    Kang, Jihoon; McCaleb, Melanie M; McLaughlin, Richard A

    2013-11-15

    High levels of turbidity and fine suspended sediments are often found in stormwater discharges from construction sites even when best management practices (BMPs) for sediment control are in place. This study evaluated turbidity reduction by three check dam types: 1) rock check dam representing a standard BMP, 2) excelsior wattle representing a fiber check dam (FCD), and 3) rock check dam wrapped with excelsior erosion control blanket (rock + excelsior ECB) representing an alternative FCD. Three check dams (all same type) were installed in a lined, 24-m ditch on a 5-7% slope and three consecutive simulated stormwater flows were run in the ditch. Additional tests were performed by adding granular polyacrylamide (PAM) on the check dams in the same manner using two sediment sources differing in clay content. Without PAM treatment, significantly higher effluent turbidity (>900 nephelometric turbidity units (NTU)) exited the ditch with rock check dams than with excelsior wattles or rock + excelsior ECBs (<440 NTU). The extent of sediment deposition between the check dam types was in the order of excelsior wattle > rock + excelsior ECB > rock check dam, indicating better water pooling behind the wattle. The PAM treatment reduced turbidity substantially (>75% relative to no PAM treatment) for all check dam types and it was very effective in excelsior wattles (<57 NTU) and rock + excelsior ECBs (<90 NTU) even during the third storm event. This study demonstrates that the passive treatment of runoff with PAM on FCDs (or rock + excelsior ECB) in construction site ditches can be very effective for sediment retention and turbidity reduction.

  4. One-step preparation of hybrid materials of polyacrylamide networks and gold nanoparticles.

    PubMed

    Song, Yonghai; Li, Zhiqiang; Wang, Li; Yao, Yong; Chen, Chuangye; Cui, Kang

    2008-06-01

    Hybrid materials of polyacrylamide networks and gold nanoparticles were prepared by directly heating an aqueous solution containing HAuCl(4), acrylamide, N,N'-methylenebis-acrylamide, and sodium sulfite (Na(2)SO(3)). Acrylamide, N,N'-methylenebis-acrylamide, and Na(2)SO(3) were used as monomers, crosslinking agent, and initiator, respectively. In the process of polyacrylamide network synthesis, HAuCl(4) was reduced by acrylamide and Na(2)SO(3) into gold nanoparticles and adsorbed on the produced polyacrylamide networks. Transmission electron microscopy proved that the size of gold nanoparticles was in the range of 3-10 nm. Atomic force microscopy showed that the gold nanoparticles homogeneously dispersed into the polyacrylamide networks matrix. The hybrid materials as absorbents may be useful in healthcare, communication technology, building industry, chromatography, water purification, and agriculture.

  5. A gel-based solid-phase amplification and its application for SNP typing and sequencing on-chip.

    PubMed

    Huang, Huan; Xiao, Pengfeng; Qi, Zongtai; Bu, Ying; Liu, Wenbo; Zhou, Guohua

    2009-12-01

    As conventional solid-phase amplification (SPA) on a two-dimensional slide has a low amplification capacity due to a limited amount of immobilized primers, we propose a three-dimensional SPA by immobilizing primers in hydrogel attached to a slide. One of the PCR primers, modified with an acrylamide group at the 5'-terminal, was copolymerized with both polyacrylamide gel and an acryl-modified glass slide, resulting in a high amplification capacity. The immobilization process was carried out by adding the catalysis reagent N,N,N',N'-tetramethylethylenediamine (TEMED) volatilized in vacuum, with uniform sample-concentration and gel-viscosity in the course of one-step nucleic acid immobilization. The porous structure of polyacrylamide gel, which allows PCR reagents such as Taq DNA polymerase, primers, dNTPs and DNA templates to freely enter the gel matrix, provides a homogeneous solution-mimicking environment for SPA on the interface or the inside of gel pads. Based on gel-based SPA, genotypes of different samples were accurately discriminated by either dual-color fluorescence hybridization or BAMPER (Bioluminometric Assay coupled with Modified Primer Extension Reactions). Pyrosequencing was also successfully carried out on SPA products. As the linkage between DNA molecules and gel is very strong, SPA products immobilized on gel pads could be reused several times if extended strands were removed by electrophoresis. Thus, the gel-based SPA provides a powerful tool for directly using on-chip amplicons for parallel detection.

  6. Chemical gel barriers as low-cost alternative to containment and in situ cleanup of hazardous wastes to protect groundwater

    SciTech Connect

    1997-01-01

    Chemical gel barriers are being considered as a low-cost alternative for containment and in situ cleanup of hazardous wastes to protect groundwater. Most of the available gels in petroleum application are non-reactive and relative impermeable, providing a physical barriers for all fluids and contaminants. However, other potential systems can be envisioned. These systems could include gels that are chemically reactive and impermeable such that most phase are captured by the barriers but the contaminants could diffuse through the barriers. Another system that is chemically reactive and permeable could have potential applications in selectivity capturing contaminants while allowing water to pass through the barriers. This study focused on chemically reactive and permeable gel barriers. The gels used in experiment are DuPont LUDOX SM colloidal silica gel and Pfizer FLOPAAM 1330S hydrolyzed polyacrylamide (HPAM) gel.

  7. Solids and nutrient removal from flushed swine manure using polyacrylamides

    SciTech Connect

    Vanotti, M.B.; Hunt, P.G.

    1999-12-01

    Most of the organic nutrients and reduced carbon (C) materials in liquid swine manure are contained in fine suspended particles that are not separated by available mechanical separators. Treatment with polyacrylamide (PAM) polymers prior to mechanical removal or gravity settling has the potential for enhancing solids-liquid separation, thus concentrating nitrogen (N), phosphorus (P), and organic C. In this work, the authors determined PAM charge and density characteristics most desirable for swine wastewater applications and established the optimum chemical requirement. Treatments were applied to flushed manure from two swine operations in North Carolina. Cationic PAMs significantly increased solids separation while performance of neutral and anionic types was not different from a control. Cationic PAMs with moderate-charge density (20%) were more effective than polymers with higher charge density. Flocs were large and effectively retained with a 1-mm screen. Optimum PAM rate varied with the amount of total suspended solids (TSS) in the liquid manure; 26 and 79 mg PAM/L for samples containing 1.5 and 4.1 g TSS/L, respectively. Corresponding TSS removal efficiencies were 90 to 94%. In contrast, screening without PAM treatment captured only 5 to 14% of the suspended solids. Polymer usage rate was consistent and averaged 2.0{degree} based on weight of dry solids produced. Volatile suspended solids (VSS) were highly correlated with TSS and comprised 79.5% of TSS. Chemical oxygen demand (COD) and organic nutrient concentrations in the effluent were also significantly decreased by PAM treatment. The decrease of COD concentration, an important consideration for odor control, was linearly related with removal of suspended solids, at a rate of 2.0 g COD/g TSS and 2.6 g COD/g VSS. Removal efficiency of organic N and P followed approximately a 1:1 relationship with removal efficiency of TSS. Chemical cost to capture 90% of the suspended solids was estimated to be $0.026 per

  8. A comparative in vitro study of the digestibility of heat- and high pressure-induced gels prepared from industrial milk whey proteins

    NASA Astrophysics Data System (ADS)

    He, Jin-Song; Mu, Tai-Hua; Wang, Juan

    2013-06-01

    We undertook this study to compare the digestibility of heat- and high pressure-induced gels produced from whey protein isolate (WPI). To simulate in vivo gastrointestinal digestion of WPI gels, a pepsin-trypsin digestion system was used. The in vitro protein digestibility of WPI gels induced by high pressure (400 MPa and 30 min; P-gel) and those induced by heat (80°C and 30 min; H-gel) was compared using a protein concentration of 0.14 g mL-1. The in vitro protein digestibility of P-gels was significantly greater than that of H-gels (p<0.05). The size-exclusion chromatography profiles of the hydrolysates showed that the P-gel generated more and smaller peptides than natural WPI and H-gels. Furthermore, Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed some soluble disulfide-mediated aggregation in the P-gel, while there was more insoluble aggregation in the H-gel than the P-gel. The P-gel was more sensitive to proteinase than the H-gel, which was related to the content of S-S bonds, and this in turn could be attributed to the differences in the gelation mechanism between the H-gel and P-gel.

  9. Materials Science and Device Physics of 2-Dimensional Semiconductors

    NASA Astrophysics Data System (ADS)

    Fang, Hui

    Materials and device innovations are the keys to future technology revolution. For MOSFET scaling in particular, semiconductors with ultra-thin thickness on insulator platform is currently of great interest, due to the potential of integrating excellent channel materials with the industrially mature Si processing. Meanwhile, ultra-thin thickness also induces strong quantum confinement which in turn affect most of the material properties of these 2-dimensional (2-D) semiconductors, providing unprecedented opportunities for emerging technologies. In this thesis, multiple novel 2-D material systems are explored. Chapter one introduces the present challenges faced by MOSFET scaling. Chapter two covers the integration of ultrathin III V membranes with Si. Free standing ultrathin III-V is studied to enable high performance III-V on Si MOSFETs with strain engineering and alloying. Chapter three studies the light absorption in 2-D membranes. Experimental results and theoretical analysis reveal that light absorption in the 2-D quantum membranes is quantized into a fundamental physical constant, where we call it the quantum unit of light absorption, irrelevant of most of the material dependent parameters. Chapter four starts to focus on another 2-D system, atomic thin layered chalcogenides. Single and few layered chalcogenides are first explored as channel materials, with focuses in engineering the contacts for high performance MOSFETs. Contact treatment by molecular doping methods reveals that many layered chalcogenides other than MoS2 exhibit good transport properties at single layer limit. Finally, Chapter five investigated 2-D van der Waals heterostructures built from different single layer chalcogenides. The investigation in a WSe2/MoS2 hetero-bilayer shows a large Stokes like shift between photoluminescence peak and lowest absorption peak, as well as strong photoluminescence intensity, consistent with spatially indirect transition in a type II band alignment in this

  10. Comparative Skeletal Muscle Proteomics Using Two-Dimensional Gel Electrophoresis

    PubMed Central

    Murphy, Sandra; Dowling, Paul; Ohlendieck, Kay

    2016-01-01

    The pioneering work by Patrick H. O’Farrell established two-dimensional gel electrophoresis as one of the most important high-resolution protein separation techniques of modern biochemistry (Journal of Biological Chemistry 1975, 250, 4007–4021). The application of two-dimensional gel electrophoresis has played a key role in the systematic identification and detailed characterization of the protein constituents of skeletal muscles. Protein changes during myogenesis, muscle maturation, fibre type specification, physiological muscle adaptations and natural muscle aging were studied in depth by the original O’Farrell method or slightly modified gel electrophoretic techniques. Over the last 40 years, the combined usage of isoelectric focusing in the first dimension and sodium dodecyl sulfate polyacrylamide slab gel electrophoresis in the second dimension has been successfully employed in several hundred published studies on gel-based skeletal muscle biochemistry. This review focuses on normal and physiologically challenged skeletal muscle tissues and outlines key findings from mass spectrometry-based muscle proteomics, which was instrumental in the identification of several thousand individual protein isoforms following gel electrophoretic separation. These muscle-associated protein species belong to the diverse group of regulatory and contractile proteins of the acto-myosin apparatus that forms the sarcomere, cytoskeletal proteins, metabolic enzymes and transporters, signaling proteins, ion-handling proteins, molecular chaperones and extracellular matrix proteins. PMID:28248237

  11. Comparative Skeletal Muscle Proteomics Using Two-Dimensional Gel Electrophoresis.

    PubMed

    Murphy, Sandra; Dowling, Paul; Ohlendieck, Kay

    2016-09-09

    The pioneering work by Patrick H. O'Farrell established two-dimensional gel electrophoresis as one of the most important high-resolution protein separation techniques of modern biochemistry (Journal of Biological Chemistry1975, 250, 4007-4021). The application of two-dimensional gel electrophoresis has played a key role in the systematic identification and detailed characterization of the protein constituents of skeletal muscles. Protein changes during myogenesis, muscle maturation, fibre type specification, physiological muscle adaptations and natural muscle aging were studied in depth by the original O'Farrell method or slightly modified gel electrophoretic techniques. Over the last 40 years, the combined usage of isoelectric focusing in the first dimension and sodium dodecyl sulfate polyacrylamide slab gel electrophoresis in the second dimension has been successfully employed in several hundred published studies on gel-based skeletal muscle biochemistry. This review focuses on normal and physiologically challenged skeletal muscle tissues and outlines key findings from mass spectrometry-based muscle proteomics, which was instrumental in the identification of several thousand individual protein isoforms following gel electrophoretic separation. These muscle-associated protein species belong to the diverse group of regulatory and contractile proteins of the acto-myosin apparatus that forms the sarcomere, cytoskeletal proteins, metabolic enzymes and transporters, signaling proteins, ion-handling proteins, molecular chaperones and extracellular matrix proteins.

  12. Fundamentals of gel dosimeters

    NASA Astrophysics Data System (ADS)

    McAuley, K. B.; Nasr, A. T.

    2013-06-01

    Fundamental chemical and physical phenomena that occur in Fricke gel dosimeters, polymer gel dosimeters, micelle gel dosimeters and genipin gel dosimeters are discussed. Fricke gel dosimeters are effective even though their radiation sensitivity depends on oxygen concentration. Oxygen contamination can cause severe problems in polymer gel dosimeters, even when THPC is used. Oxygen leakage must be prevented between manufacturing and irradiation of polymer gels, and internal calibration methods should be used so that contamination problems can be detected. Micelle gel dosimeters are promising due to their favourable diffusion properties. The introduction of micelles to gel dosimetry may open up new areas of dosimetry research wherein a range of water-insoluble radiochromic materials can be explored as reporter molecules.

  13. DEVELOPMENT OF POLYMER GEL SYSTEMS TO IMPROVE VOLUMETRIC SWEEP AND REDUCE PRODUCING WATER/OIL RATIOS

    SciTech Connect

    G. Paul Willhite; Stan McCool; Don W. Green; Min Cheng; Rajeev Jain; Tuan Nguyen

    2003-11-01

    Gelled polymer treatments are applied to oil reservoirs to increase oil production and to reduce water production by altering the fluid movement within the reservoir. This report describes the results of the first year of a three-year research program that is aimed at the understanding of the chemistry of gelation and the fundamental mechanisms that alter the flows of oil and water in reservoir rocks after a gel treatment. Work has focused on a widely-applied system in field applications, the partially hydrolyzed polyacrylamide-chromium acetate gel. Gelation occurs by network formation through the crosslinking of polyacrylamide molecules as a result of reaction with chromium acetate. The initial reaction between chromium acetate and one polymer is referred to as the uptake reaction. The uptake reaction was studied as functions of chromium and polymer concentrations and pH values. Experimental data were regressed to determine a rate equation that describes the uptake reaction of chromium by polyacrylamide. Pre-gel aggregates form and grow as the reactions between chromium acetate and polyacrylamide proceed. A statistical model that describes the growth of pre-gel aggregates was developed using the theory of branching processes. The model gives molecular weight averages that are expressed as functions of the conversion of the reactive sites on chromium acetate or on the polymer molecule. Results of the application of the model correlate well with experimental data of viscosity and weight-average molecular weight and gives insights into the gelation process. A third study addresses the flow of water and oil in rock material after a gel treatment. Previous works have shown that gel treatments usually reduce the permeability to water to a greater extent than the permeability to oil is reduced. This phenomenon is referred to as disproportionate permeability reduction (DPR). Flow experiments were conducted to determine the effect of polymer and chromium concentrations on

  14. Enhanced removal of methylene blue and methyl violet dyes from aqueous solution using a nanocomposite of hydrolyzed polyacrylamide grafted xanthan gum and incorporated nanosilica.

    PubMed

    Ghorai, Soumitra; Sarkar, Asish; Raoufi, Mohammad; Panda, Asit Baran; Schönherr, Holger; Pal, Sagar

    2014-04-09

    The synthesis and characterization of a novel nanocomposite is reported that was developed as an efficient adsorbent for the removal of toxic methylene blue (MB) and methyl violet (MV) from aqueous solution. The nanocomposite comprises hydrolyzed polyacrylamide grafted onto xanthan gum as well as incorporated nanosilica. The synthesis exploits the saponification of the grafted polyacrylamide and the in situ formation of nanoscale SiO2 by a sol-gel reaction, in which the biopolymer matrix promotes the silica polymerization and therefore acts as a novel template for nanosilica formation. The detailed investigation of the kinetics and the adsorption isotherms of MB and MV from aqueous solution showed that the dyes adsorb rapidly, in accordance with a pseudo-second-order kinetics and a Langmuir adsorption isotherm. The entropy driven process was furthermore found to strongly depend on the point of zero charge (pzc) of the adsorbent. The remarkably high adsorption capacity of dyes on the nanocomposites (efficiency of MB removal, 99.4%; maximum specific removal Qmax, 497.5 mg g(-1); and efficiency of MV removal, 99.1%; Qmax, 378.8 mg g(-1)) is rationalized on the basis of H-bonding interactions as well as dipole-dipole and electrostatic interactions between anionic adsorbent and cationic dye molecules. Because of the excellent regeneration capacity the nanocomposites are considered interesting materials for the uptake of, for instance, toxic dyes from wastewater.

  15. Rapid (ten-minute) pore-gradient electrophoresis of proteins and peptides in Micrograd gels.

    PubMed

    Wrigley, C W; Margolis, J

    1992-01-01

    Precast gradient gels of short migration length (25 mm) have been developed to provide rapid electrophoretic separation without loss of resolution. These Micrograd gels have been prepared in gel ranges (conventional and unique) to match pore-gradient electrophoresis conditions to proteins/peptides ranging in size from several hundreds to millions. The Hylinx Micrograd gel combines an extreme gel range (6 to 48% polyacrylamide) with a novel crosslinker to provide sieving of polypeptides, and pore-limit electrophoresis of the smallest proteins (e.g. insulin monomer). All gel ranges (such as 3 to 30%) provide zone sharpening in routine analysis of conventional protein mixtures (e.g. serum) within 10 min electrophoresis at 200 to 300 volts. The gels are thin (1 mm) and thus stain quickly, but the gel cassette is of conventional overall width (83 mm), thus fitting many apparatus designs and accommodating 12 samples. The gels are finding valuable use in screening applications, requiring the electrophoretic analysis of many samples, and in cases where a rapid answer is needed, such as monitoring protein purification. The gels have proved particularly useful, in-house, for the latter application in developing Gradipore's new large-scale preparative electrophoresis system, the Gradiflow.

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

    SciTech Connect

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

    1989-04-25

    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 and 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.

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

    SciTech Connect

    Zhang, Jian-Shi; Giometti, Carol S.; Tollaksen, Sandra L.

    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.

  18. Database of two-dimensional polyacrylamide gel electrophoresis of proteins labeled with CyDye DIGE Fluor saturation dye.

    PubMed

    Fujii, Kazuyasu; Kondo, Tadashi; Yokoo, Hideki; Okano, Tetsuya; Yamada, Masayo; Yamada, Tesshi; Iwatsuki, Keiji; Hirohashi, Setsuo

    2006-03-01

    CyDye DIGE Fluor saturation dye (saturation dye, GE Healthcare Amersham Biosciences) enables highly sensitive 2-D PAGE. As the dye reacts with all reduced cysteine thiols, 2-D PAGE can be performed with a lower amount of protein, compared with CyDye DIGE Fluor minimal dye (GE Healthcare Amersham Biosciences), the sensitivity of which is equivalent to that of silver staining. We constructed a 2-D map of the saturation dye-labeled proteins of a liver cancer cell line (HepG2) and identified by MS 92 proteins corresponding to 123 protein spots. Functional classification revealed that the identified proteins had chaperone, protein binding, nucleotide binding, metal ion binding, isomerase activity, and motor activity. The functional distribution and the cysteine contents of the proteins were similar to those in the most comprehensive 2-D database of hepatoma cells (Seow et al.., Electrophoresis 2000, 21, 1787-1813), where silver staining was used for protein visualization. Hierarchical clustering on the basis of the quantitative expression profiles of the 123 characterized spots labeled with two charge- and mass-matched saturation dyes (Cy3 and Cy5) discriminated between nine hepatocellular carcinoma cell lines and primary cultured hepatocytes from five individuals, suggesting the utility of saturation dye and our database for proteomic studies of liver cancer.

  19. Microfluidic device having an immobilized pH gradient and PAGE gels for protein separation and analysis

    DOEpatents

    Sommer, Gregory J.; Hatch, Anson V.; Singh, Anup K.; Wang, Ying-Chih

    2012-12-11

    Disclosed is a novel microfluidic device enabling on-chip implementation of a two-dimensional separation methodology. Previously disclosed microscale immobilized pH gradients (IPG) are combined with perpendicular polyacrylamide gel electrophoresis (PAGE) microchannels to achieve orthogonal separations of biological samples. Device modifications enable inclusion of sodium dodecyl sulfate (SDS) in the second dimension. The device can be fabricated to use either continuous IPG gels, or the microscale isoelectric fractionation membranes we have also previously disclosed, for the first dimension. The invention represents the first all-gel two-dimensional separation microdevice, with significantly higher resolution power over existing devices.

  20. Microfluidic device having an immobilized pH gradient and page gels for protein separation and analysis

    DOEpatents

    Sommer, Gregory J; Hatch, Anson V; Singh, Anup K; Wang, Ying-Chih

    2014-05-20

    Disclosed is a novel microfluidic device enabling on-chip implementation of a two-dimensional separation methodology. Previously disclosed microscale immobilized pH gradients (IPG) are combined with perpendicular polyacrylamide gel electrophoresis (PAGE) microchannels to achieve orthogonal separations of biological samples. Device modifications enable inclusion of sodium dodecyl sulfate (SDS) in the second dimension. The device can be fabricated to use either continuous IPG gels, or the microscale isoelectric fractionation membranes we have also previously disclosed, for the first dimension. The invention represents the first all-gel two-dimensional separation microdevice, with significantly higher resolution power over existing devices.

  1. Some considerations concerning four-ball machine testing of the polyacrylamide solutions

    NASA Astrophysics Data System (ADS)

    Drumeanu, A. C.

    2017-02-01

    Polyacrylamide (PAM) is one of the most widely used and technically important water-soluble polymers. Polyacrylamide (PAM) is usually obtained by free radical polymerization of acrylamide (AM) and in its partially hydrolysed form is a synthetic straight-chain polymer of acrylamide monomers, some of which have been hydrolysed. The structure of HPAM molecule is a flexible chain. This kind of structure is known as a random coil in polymer chemistry. Due to the hydrolysed groups contained in its molecule, HPAM has multiple charges distributed along the chain that make it a polyelectrolyte. The paper presents the experimental results concerning the lubricant solutions based on polyacrylamide behaviour when were tested on the four ball machine. It has to be mentioned that this kind of polymer was not used until now in lubrication and the studies concerning its tribological behaviour are at the beginning.

  2. Protein differences between normal and oligospermic human sperm demonstrated by two-dimensional gel electrophoresis.

    PubMed

    Morgentaler, A; Schopperle, W M; Crocker, R H; DeWolf, W C

    1990-11-01

    Protein expression by sperm obtained from men with normal semen analysis and men with oligospermia were evaluated by two-dimensional gel electrophoresis. Proteins were solubilized in a 9.5 M urea/2% Nonidet-P40 (LKB, Bromma, Sweden) lysis buffer and underwent second dimension separation on 10 to 16% polyacrylamide gradient gels. A set of 36 invariant proteins was identified in all normospermic samples, whereas 8 of 10 evaluable oligospermic samples lacked 1 or more of the invariant proteins. Proteins absent in oligospermic samples may be critical to normal sperm function and may serve as markers for infertility.

  3. Orthogonally bifunctionalised polyacrylamide nanoparticles: a support for the assembly of multifunctional nanodevices

    NASA Astrophysics Data System (ADS)

    Giuntini, F.; Dumoulin, F.; Daly, R.; Ahsen, V.; Scanlan, E. M.; Lavado, A. S. P.; Aylott, J. W.; Rosser, G. A.; Beeby, A.; Boyle, R. W.

    2012-03-01

    Polyacrylamide nanoparticles bearing two orthogonal reactive functionalities were prepared by reverse microemulsion polymerisation. Water-soluble photosensitisers and peptide or carbohydrate moieties were sequentially attached to the new nanospecies by orthogonal conjugations based on copper-catalysed azide-alkyne cycloaddition and isothiocyanate chemistry.Polyacrylamide nanoparticles bearing two orthogonal reactive functionalities were prepared by reverse microemulsion polymerisation. Water-soluble photosensitisers and peptide or carbohydrate moieties were sequentially attached to the new nanospecies by orthogonal conjugations based on copper-catalysed azide-alkyne cycloaddition and isothiocyanate chemistry. Electronic supplementary information (ESI) available. See DOI: 10.1039/c2nr11947a

  4. Evaluation of a polyacrylamide soil additive to reduce agricultural-associated contamination.

    PubMed

    Krauth, D M; Bouldin, J L; Green, V S; Wren, P S; Baker, W H

    2008-08-01

    Polyacrylamide is an effective water treatment product used to reduce suspended sediment and associated contaminants. An anionic polyacrylamide-containing product was tested for sediment and associated contaminant reduction and potential toxicity in agricultural irrigation and rainfall runoff. The product effectively reduced turbidity, total suspended solids, and phosphate concentrations in the field when compared to the untreated runoff waters. Acute survival of Ceriodaphnia dubia and Pimephales promelas was not decreased compared to laboratory controls. No significant increases in toxicity were measured in 10-d sediment toxicity tests with Chironomus dilutus. Product applications were effective in controlling sediment and nutrient contamination without increasing measured toxicity.

  5. Microplate array diagonal gel electrophoresis for cohort studies of microsatellite loci.

    PubMed

    Chen, Xiao-he; O'Dell, Sandra D; Day, Ian N M

    2002-05-01

    After PCR amplification, we have achieved precise sizing of trinucleotide and tetranucleotide microsatellite alleles on 96-well open-faced polyacrylamide microplate array diagonal gel electrophoresis (MADGE) gels: two tetranucleotide repeats, HUMTHOI (five alleles 248-263 bp) and DYS390 (eight alleles 200-228 bp), and DYS392, a trinucleotide repeat (eight alleles 210-231 bp). A gel matrix of Duracryl, a high mechanical strength polyacrylamide derivative, and appropriate ionic conditions provide the 1.3%-1.5% band resolution required. No end-labeling of primers is needed, as the sensitive Vistra Green intercalating dye is used for the visualization of bands. Co-run markers bracketing the PCR fragments ensure accurate sizing without inter-lane variability. Electrophoresis of multiple gels in a thermostatically controlled tank allows up to 1000 samples to be run in 90 min. Gel images were analyzed using a Fluorlmager 595 fluorescent scanning system, and alleles were identified using Phoretix software for band migration measurement and Microsoft Excel to compute fragment sizes. Estimated sizes were interpolated precisely to achieve accurate binning. Microsatellite-MADGE represents a utilitarian methodfor high-throughput genotyping in cohort studies, using standard laboratory equipment.

  6. Fluid diversion and sweep improvement with chemical gels in oil recovery processes. Final report

    SciTech Connect

    Seright, R.S.; Martin, F.D.

    1992-09-01

    The objectives of this project were to identify the mechanisms by which gel treatments divert fluids in reservoirs and to establish where and how gel treatments are best applied. Several different types of gelants were examined, including polymer-based gelants, a monomer-based gelant, and a colloidal-silica gelant. This research was directed at gel applications in water injection wells, in production wells, and in high-pressure gas floods. The work examined how the flow properties of gels and gelling agents are influenced by permeability, lithology, and wettability. Other goals included determining the proper placement of gelants, the stability of in-place gels, and the types of gels required for the various oil recovery processes and for different scales of reservoir heterogeneity. During this three-year project, a number of theoretical analyses were performed to determine where gel treatments are expected to work best and where they are not expected to be effective. The most important, predictions from these analyses are presented. Undoubtedly, some of these predictions will be controversial. However, they do provide a starting point in establishing guidelines for the selection of field candidates for gel treatments. A logical next step is to seek field data that either confirm or contradict these predictions. The experimental work focused on four types of gels: (1) resorcinol-formaldehyde, (2) colloidal silica, (3) Cr{sup 3+}(chloride)-xanthan, and (4) Cr{sup 3+}(acetate)-polyacrylamide. All experiments were performed at 41{degrees}C.

  7. Combining high-throughput MALDI-TOF mass spectrometry and isoelectric focusing gel electrophoresis for virtual 2D gel-based proteomics.

    PubMed

    Lohnes, Karen; Quebbemann, Neil R; Liu, Kate; Kobzeff, Fred; Loo, Joseph A; Ogorzalek Loo, Rachel R

    2016-07-15

    The virtual two-dimensional gel electrophoresis/mass spectrometry (virtual 2D gel/MS) technology combines the premier, high-resolution capabilities of 2D gel electrophoresis with the sensitivity and high mass accuracy of mass spectrometry (MS). Intact proteins separated by isoelectric focusing (IEF) gel electrophoresis are imaged from immobilized pH gradient (IPG) polyacrylamide gels (the first dimension of classic 2D-PAGE) by matrix-assisted laser desorption/ionization (MALDI) MS. Obtaining accurate intact masses from sub-picomole-level proteins embedded in 2D-PAGE gels or in IPG strips is desirable to elucidate how the protein of one spot identified as protein 'A' on a 2D gel differs from the protein of another spot identified as the same protein, whenever tryptic peptide maps fail to resolve the issue. This task, however, has been extremely challenging. Virtual 2D gel/MS provides access to these intact masses. Modifications to our matrix deposition procedure improve the reliability with which IPG gels can be prepared; the new procedure is described. Development of this MALDI MS imaging (MSI) method for high-throughput MS with integrated 'top-down' MS to elucidate protein isoforms from complex biological samples is described and it is demonstrated that a 4-cm IPG gel segment can now be imaged in approximately 5min. Gel-wide chemical and enzymatic methods with further interrogation by MALDI MS/MS provide identifications, sequence-related information, and post-translational/transcriptional modification information. The MSI-based virtual 2D gel/MS platform may potentially link the benefits of 'top-down' and 'bottom-up' proteomics.

  8. Sol-Gel Glasses

    NASA Technical Reports Server (NTRS)

    Mukherjee, S. P.

    1985-01-01

    Multicomponent homogeneous, ultrapure noncrystalline gels/gel derived glasses are promising batch materials for the containerless glass melting experiments in microgravity. Hence, ultrapure, homogeneous gel precursors could be used to: (1) investigate the effect of the container induced nucleation on the glass forming ability of marginally glass forming compositions; and (2) investigate the influence of gravity on the phase separation and coarsening behavior of gel derived glasses in the liquid-liquid immiscibility zone of the nonsilicate systems having a high density phase. The structure and crystallization behavior of gels in the SiO2-GeO2 as a function of gel chemistry and thermal treatment were investigated. As are the chemical principles involved in the distribution of a second network former in silica gel matrix being investigated. The procedures for synthesizing noncrystalline gels/gel-monoliths in the SiO2-GeO2, GeO2-PbO systems were developed. Preliminary investigations on the levitation and thermal treatment of germania silicate gel-monoliths in the Pressure Facility Acoustic Levitator were done.

  9. Effects of Polyacrylamide and Organic Matter on Microbes associated to Soil Aggregation of Norfolk Loamy Sand

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Polyacrylamide (PAM, anionic formulation of molecular size 12 MDa and 35% charge density) has been reported to increase aggregation and improve soil physical properties in United States southeastern Coastal Plain loamy sand soils, but nothing is known about the effects of PAM on microbes associated ...

  10. Soil aggregate stability as affected by clay mineralogy and polyacrylamide addition

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The addition of polyacrylamide (PAM) to soil leads to stabilization of existing aggregates and improved bonding between, and aggregation of adjacent soil particles However, the dependence of PAM efficacy as an aggregate stabilizing agent on soil-clay mineralogy has not been studied. Sixteen soil sam...

  11. Polyacrylamide treatments for reducing seepage in soil-lined reservoirs: A field evaluation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Irrigation water supplies are becoming limited and there is a need to extend the usefulness of current water resources. Previous laboratory studies demonstrated that certain water soluble polyacrylamide solution (WSPAM) and cross-linked PAM granule (XPAM) treatments effectively reduced infiltration...

  12. Polyacrylamide Molecular Weight and Phosphogypsum Effects on Infiltration and Erosion in Semi-Arid Soils

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Seal formation at the surface of semi-arid soils during rainstorms reduces soil infiltration rate (IR) and causes runoff and erosion. Surface application of dry anionic polyacrylamide (PAM) with high molecular weight (MW) has been found to be effective in stabilizing soil aggregates, and decreasing ...

  13. Polyacrylamide molecular weight and phosphogypsum effects on infiltration and erosion in semi-arid soils

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Seal formation at the surface of semi-arid soils during rainstorms reduces soil infiltration rate (IR) and causes runoff and erosion. Surface application of dry anionic polyacrylamide (PAM) with high molecular weight (MW) has been found to be effective in stabilizing soil aggregates, and decreasing ...

  14. Polyacrylamide effects on aggregate and structure stability of soils with different clay mineralogy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Adding anionic polyacrylamide (PAM) to soils stabilizes existing aggregates and improves bonding between and aggregation of soil particles. However, the dependence of PAM efficacy as an aggregate stabilizing agent with soils having different clay mineralogy has not been studied. Sixteen soil samples...

  15. Polyacrylamide and biopolymer effects on flocculation, aggregate stability, and water seepage in a silt loam

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Researcher’s seek a more renewable and natural alternative for water soluble anionic polyacrylamide (PAM), a highly-effective, petroleum-derived polymer used in agriculture to control erosion and reduce water seepage from unlined irrigation structures. This study evaluated two anionic polymers: a ba...

  16. Molecular Understanding and Structural-Based Design of Polyacrylamides and Polyacrylates as Antifouling Materials.

    PubMed

    Chen, Hong; Zhao, Chao; Zhang, Mingzhen; Chen, Qiang; Ma, Jie; Zheng, Jie

    2016-04-12

    Design and synthesis of highly bioinert and biocompatible antifouling materials are crucial for a broad range of biomedical and engineering applications. Among antifouling materials, polyacrylamides and polyacrylates have proved so promising because of cheap raw materials, ease of synthesis and applicability, and abundant functional groups. The strong surface hydration and the high surface packing density of polyacrylamides and polyacrylates are considered to be the key contributors to their antifouling property. In this article, we review our studies on the design and synthesis of a series of polyacrylamides and polyacrylates with different molecular structures. These polymers can be fabricated into different architectural forms (brushes, nanoparticles, nanogels, and hydrogels), all of which are highly resistant to the attachment of proteins, cells, and bacteria. We find that small structural changes in the polymers can lead to large enhancement in surface hydration and antifouling performance, both showing a positive correlation. This reveals a general design rule for effective antifouling materials. Furthermore, polyacrylamides and polyacrylates are readily functionalized with other bioactive compounds to achieve different new multifunctionalities.

  17. Comparison of Cationic and Unmodified Starches in Reactive Extrusion of Starch-Polyacrylamide Graft Copolymers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Graft copolymers of starch and polyacrylamide (PAAm) were prepared using reactive extrusion in a corotating twin screw extruder. The effect of cationic starch modification was examined using unmodified and cationic dent starch (approximately 23% amylose) and waxy maize starch (approximately 2% amyl...

  18. Development of Polymer Gel Systems to Improve Volumetric Sweep and Reduce Producing Water/Oil Ratios

    SciTech Connect

    G. Paul Willhite; Stan McCool; Don W. Green; Min Cheng; Feiyan Chen

    2005-04-03

    Gelled polymer treatments are applied to oil reservoirs to increase oil production and to reduce water production by altering the fluid movement within the reservoir. This report describes the results of the third year of a 42 month research program that is aimed at an understanding of gelation chemistry and the fundamental mechanisms that alter the flows of oil and water in reservoir rocks after a gel treatment. Work focused on a widely applied system in the field, the partially hydrolyzed polyacrylamide-chromium acetate gel. Gelation occurs by network formation through the crosslinking of polyacrylamide molecules as a result of reaction with chromium acetate. Pre-gel aggregates form and grow as reactions between chromium acetate and polyacrylamide proceed. A mathematical model that describes uptake and crosslinking reactions as a function of time was derived. The model was probability based and provides molecular-weight averages and molecular-weight distributions of the pre-gel aggregates as a function of time and initial system conditions. A liquid chromatography apparatus to experimentally measure the size and molecular weight distributions of polymer samples was developed. The method worked well for polymer samples without the chromium crosslinker. Sample retention observed during measurements of gelant samples during the gelation process compromised the results. Other methods will be tested to measure size distributions of the pre-gel aggregates. Dissolution of carbonate minerals during the injection of gelants causes the pH of the gelant to increase. Chromium precipitates from solution at the higher pH values robbing the gelant of crosslinker. Experimental data on the transport of chromium acetate solutions through dolomite cores were obtained. A mathematical model that describes the transport of brine and chromium acetate solutions through rocks containing carbonate minerals was used to simulate the experimental results.

  19. Hydrous manganese oxide doped gel probe sampler for measuring in situ reductive dissolution rates. 1. Laboratory development.

    PubMed

    Farnsworth, Claire E; Hering, Janet G

    2010-01-01

    Reductive dissolution of redox-sensitive minerals such as manganese (Mn) oxides in natural sediments is an important mechanism for trace element mobilization into groundwater. A gel probe sampler has been constructed to study in situ reductive dissolution of Mn oxides. The gel consists of a polyacrylamide polymer matrix doped with hydrous Mn oxide (HMO). Gel slabs are mounted into a probe, which is designed to be inserted into the sediments. The amount of Mn released from the gel by reductive dissolution is determined by comparing the amount of Mn initially embedded into the gel with the amount remaining in the gel after exposure to conditions in the sediments or, in laboratory studies, to reducing agents. In this laboratory study, the performance of the gel probes was examined using the model reductant ascorbate and the Mn-reducing bacteria Shewanella oneidensis strain MR-1. In addition, a 1-D model was used to relate the reaction rates observed for HMO embedded in gels to those for HMO in suspension. One limitation of the HMO-doped gels for assessing microbial reduction rates is that the gels prevent direct contact between the microbes and the HMO and hence preclude enzymatic reduction at the cell surface. Nonetheless, the HMO-doped gel probes offer the possibility to establish a lower bound for Mn-reduction capacity in sediments.

  20. High-frequency alternating-crossed-field gel electrophoresis with neutral or slightly charged interpenetrating networks to improve DNA separation.

    PubMed

    Boyd, B M; Prausnitz, J M; Blanch, H W

    1998-12-01

    Toward improving DNA separations, this work reports the effects of high-frequency square-wave AC fields superimposed perpendicular to the direct current (DC) separation field on DNA migration in both polyacrylamide-based interpenetrating networks (IPNs) and in agarose networks. Compared to standard polyacrylamide gels, IPNs allow the separation of larger DNA (9000 bp vs. 5000 bp at 5 V/cm). In novel polyacrylamide-based IPNs, an alternating current (AC) field of 5 Hz increased the maximum DNA size separable. This effect was extended to larger DNA sizes with increasing electric-field strength up to and apparently beyond the power supply-limited maximum electric-field strength of 48 V/cm. The orthogonal AC field also increased mobility. These two results combine to yield a reduction in separation time of up to a factor of 20 in novel polyacrylamide-based IPNs. When negatively charged acrylic-acid groups were incorporated into the IPNs, the use of the AC field changed the DNA-network interaction, which altered the size dependence of DNA mobility. In agarose gels, an AC field of 50 Hz increased the size range separable; however, there was no increase in DNA mobility. There was no change in size dependence of mobility in an AC field when the number of charged groups in the agarose network was increased. Based on results in the literature, possible mechanisms were examined for the effects of the AC field on DNA separation.

  1. High-Frequency Alternating-Crossed-Field Gel Electrophoresis WithNeutral or Slightly Charged Interpenetrating Networks to Improve DNASeparation

    SciTech Connect

    Boyd, B.; Prausnitz, J.; Blanch, H.

    1998-07-01

    Toward improving DNA separations, this work reports theeffects of high-frequency square-wave AC fields superimposedperpendicular to the direct current (DC) separation field on DNAmigration in both polyacrylamide-based interpenetrating networks (IPNs)and in agarose networks. Compared to standard polyacrylamide gels, IPNsallow the separation of larger DNA (9000 bp vs. 5000 bp at 5 V/cm). Innovel polyacrylamide-based IPNs, an alternating current (AC) field of 5Hz increased the maximum DNA size separable. This effect was extended tolarger DNA sizes with increasing electric-field strength up to andapparently beyond the power supply-limited maximum electric-fieldstrength of 48 V/cm. The orthogonal AC field also increased mobility.These two results combine to yield a reduction in separation time of upto a factor of 20 in novel polyacrylamide-based IPNs. When negativelycharged acrylic-acid groups were incorporated into the IPNs, the use ofthe AC field changed the DNA-network interaction, which altered the sizedependence of DNA mobility. In agarose gels, an AC field of 50 Hzincreased the size range separable; however, there was no increase in DNAmobility. There was no change in size dependence of mobility in an ACfield when the number of charged groups in the agarose network wasincreased. Based on results in the literature, possible mechanisms wereexamined for the effects of the AC field on DNA separation.

  2. Pulse Field Gel Electrophoresis

    PubMed Central

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

    2015-01-01

    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

  3. 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.

  4. CONFORMANCE IMPROVEMENT USING GELS

    SciTech Connect

    Randall S. Seright

    2003-09-01

    This report describes work performed during the second year of the project, ''Conformance Improvement Using Gels.'' The project has two objectives. The first objective is to identify gel compositions and conditions that substantially reduce flow through fractures that allow direct channeling between wells, while leaving secondary fractures open so that high fluid injection and production rates can be maintained. The second objective is to optimize treatments in fractured production wells, where the gel must reduce permeability to water much more than that to oil. Pore-level images from X-ray computed microtomography were re-examined for Berea sandstone and porous polyethylene. This analysis suggests that oil penetration through gel-filled pores occurs by a gel-dehydration mechanism, rather than a gel-ripping mechanism. This finding helps to explain why aqueous gels can reduce permeability to water more than to oil. We analyzed a Cr(III)-acetate-HPAM gel treatment in a production well in the Arbuckle formation. The availability of accurate pressure data before, during, and after the treatment was critical for the analysis. After the gel treatment, water productivity was fairly constant at about 20% of the pre-treatment value. However, oil productivity was stimulated by a factor of 18 immediately after the treatment. During the six months after the treatment, oil productivity gradually decreased to approach the pre-treatment value. To explain this behavior, we proposed that the fracture area open to oil flow was increased substantially by the gel treatment, followed by a gradual closing of the fractures during subsequent production. For a conventional Cr(III)-acetate-HPAM gel, the delay between gelant preparation and injection into a fracture impacts the placement, leakoff, and permeability reduction behavior. Formulations placed as partially formed gels showed relatively low pressure gradients during placement, and yet substantially reduced the flow capacity of

  5. Acrylamide-agarose copolymers: improved resolution of high molecular mass proteins in two-dimensional gel electrophoresis.

    PubMed

    Roncada, Paola; Cretich, Marina; Fortin, Riccardo; Agosti, Susanna; De Franceschi, Lucia; Greppi, Gian Franco; Turrini, Francesco; Carta, Franco; Turri, Stefano; Levi, Marinella; Chiari, Marcella

    2005-06-01

    A method was developed in order to analyse high molecular mass proteins by two-dimensional (2-D) electrophoresis using a copolymer of acrylamide and allyl agarose instead of Bis cross-linked polyacrylamide (PA) gels in sodium dodecyl sulphate-electrophoresis. In this work, the matrix composition was optimised to improve the resolution of proteins larger than 200 kDa. The new gel type does not entrap large proteins and protein complexes at the application site. Mechanical properties were investigated through rheological measurements, which suggested the formation of a highly entangled elastomeric soft gel. A high 2-D resolution of proteins, extracted from membranes of red blood cells, was obtained in these gels. An example of tryptic digestion, peptide extraction and matrix-assisted laser desorption/ionisation-time of flight mass spectrometry was reported. The results demonstrate that the new gel is fully compatible with mass spectrometry protein analysis.

  6. An Algorithm to Calculate Phase-Center Offset of Aperture Antennas when Measuring 2-Dimensional Radiation Patterns

    DTIC Science & Technology

    2015-01-01

    An Algorithm to Calculate Phase-Center Offset of Aperture Antennas when Measuring 2-Dimensional Radiation Patterns by Patrick Debroux...Offset of Aperture Antennas when Measuring 2-Dimensional Radiation Patterns Patrick Debroux and Berenice Verdin Survivability/Lethality Analysis... Antennas when Measuring 2-Dimensional Radiation Patterns 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S

  7. CONFORMANCE IMPROVEMENT USING GELS

    SciTech Connect

    Randall S. Seright

    2004-09-30

    This report describes work performed during the third and final year of the project, ''Conformance Improvement Using Gels.'' Corefloods revealed throughput dependencies of permeability reduction by polymers and gels that were much more prolonged during oil flow than water flow. This behavior was explained using simple mobility ratio arguments. A model was developed that quantitatively fits the results and predicts ''clean up'' times for oil productivity when production wells are returned to service after application of a polymer or gel treatment. X-ray computed microtomography studies of gels in strongly water-wet Berea sandstone and strongly oil-wet porous polyethylene suggested that oil penetration through gel-filled pores occurs by a gel-dehydration mechanism, rather than gel-ripping or gel-displacement mechanisms. In contrast, analysis of data from the University of Kansas suggests that the gel-ripping or displacement mechanisms are more important in more permeable, strongly water-wet sandpacks. These findings help to explain why aqueous gels can reduce permeability to water more than to oil under different conditions. Since cement is the most commonly used material for water shutoff, we considered when gels are preferred over cements. Our analysis and experimental results indicated that cement cannot be expected to completely fill (top to bottom) a vertical fracture of any width, except near the wellbore. For vertical fractures with apertures less than 4 mm, the cement slurry will simply not penetrate very far into the fracture. For vertical fractures with apertures greater than 4 mm, the slurry may penetrate a substantial distance into the bottom part of the fracture. However, except near the wellbore, the upper part of the fracture will remain open due to gravity segregation. We compared various approaches to plugging fractures using gels, including (1) varying polymer content, (2) varying placement (extrusion) rate, (3) using partially formed gels, (4

  8. CONFORMANCE IMPROVEMENT USING GELS

    SciTech Connect

    Randall S. Seright

    2004-03-01

    This technical progress report describes work performed from September 1, 2003, through February 29, 2004, for the project, ''Conformance Improvement Using Gels.'' We examined the properties of several ''partially formed'' gels that were formulated with a combination of high and low molecular weight HPAM polymers. After placement in 4-mm-wide fractures, these gels required about 25 psi/ft for brine to breach the gel (the best performance to date in fractures this wide). After this breach, stabilized residual resistance factors decreased significantly with increased flow rate. Also, residual resistance factors were up to 9 times greater for water than for oil. Nevertheless, permeability reduction factors were substantial for both water and oil flow. Gel with 2.5% chopped fiberglass effectively plugged 4-mm-wide fractures if a 0.5-mm-wide constriction was present. The ability to screen-out at a constriction appears crucial for particulate incorporation to be useful in plugging fractures. In addition to fiberglass, we examined incorporation of polypropylene fibers into gels. Once dispersed in brine or gelant, the polypropylene fibers exhibited the least gravity segregation of any particulate that we have tested to date. In fractures with widths of at least 2 mm, 24-hr-old gels (0.5% high molecular weight HPAM) with 0.5% fiber did not exhibit progressive plugging during placement and showed extrusion pressure gradients similar to those of gels without the fiber. The presence of the fiber roughly doubled the gel's resistance to first breach by brine flow. The breaching pressure gradients were not as large as for gels made with high and low molecular weight polymers (mentioned above). However, their material requirements and costs (i.e., polymer and/or particulate concentrations) were substantially lower than for those gels. A partially formed gel made with 0.5% HPAM did not enter a 0.052-mm-wide fracture when applying a pressure gradient of 65 psi/ft. This result

  9. Cytoplasmic polyhedrosis virus classification by electropherotype; validation by serological analyses and agarose gel electrophoresis.

    PubMed

    Mertens, P P; Crook, N E; Rubinstein, R; Pedley, S; Payne, C C

    1989-01-01

    Serological analyses of several different cytoplasmic polyhedrosis viruses (CPVs), including two type 1 CPVs from Bombyx mori, type 1 CPV from Dendrolimus spectabilis, type 12 CPV from Autographa gamma, type 2 CPV from Inachis io, type 5 CPV from Orgyia pseudotsugata and type 5 CPV from Heliothis armigera, demonstrated a close correlation between the antigenic properties of the polyhedrin or virus particle structural proteins and the genomic dsRNA electropherotypes. The dsRNAs of these viruses were analysed by electrophoresis in 3% and 10% polyacrylamide gels with a discontinuous Tris-HCl/Tris-glycine buffer system or by 1% agarose gel electrophoresis using a continuous Tris-acetate-EDTA buffer system. Electrophoretic analysis in agarose gels was found to be the most suitable for the classification of CPV isolates into electropherotypes, and the results obtained showed a close correlation with the observed antigenic relationships between different virus isolates. However, electrophoretic analysis in 10% polyacrylamide gels was most sensitive for the detection of intra-type variation and the presence of mixed virus isolates.

  10. Amended final report on the safety assessment of polyacrylamide and acrylamide residues in cosmetics.

    PubMed

    2005-01-01

    Polyacrylamide is a polymer of controllable molecular weight formed by the polymerization of acrylamide monomers available in one of three forms: solid (powder or micro beads), aqueous solution, or inverse emulsions (in water droplets coated with surfactant and suspended in mineral oil). Residual acrylamide monomer is likely an impurity in most Polyacrylamide preparations, ranging from <1 ppm to 600 ppm. Higher levels of acrylamide monomers are present in the solid form compared to the other two forms. Polyacrylamide is reportedly used in 110 cosmetic formulations, at concentrations ranging from 0.05% to 2.8%. Residual levels of acrylamide in Polyacrylamide can range from <.01% to 0.1%, although representative levels were reported at 0.02% to 0.03%. Because of the large sizes of Polyacrylamide polymers, they do not penetrate the skin. Polyacrylamide itself is not significantly toxic. For example, an acute oral toxicity study of Polyacrylamide in rats reported that a single maximum oral dose of 4.0 g/kg body weight was tolerated. In subchronic oral toxicity studies, rats and dogs treated with Polyacrylamide at doses up to 464 mg/kg body weight showed no signs of toxicity. Several 2-year chronic oral toxicity studies in rats and dogs fed diets containing up to 5% Polyacrylamide had no significant adverse effects. Polyacrylamide was not an ocular irritant in animal tests. No compound-related lesions were noted in a three-generation reproductive study in which rats were fed 500 or 2000 ppm Polyacrylamide in their diet. Polyacrylamide was not carcinogenic in several chronic animal studies. Human cutaneous tolerance tests performed to evaluate the irritation of 5% (w/w) Polyacrylamide indicated that the compound was well tolerated. Acrylamide monomer residues do penetrate the skin. Acrylamide tested in a two-generation reproductive study at concentrations up to 5 mg/kg day(- 1) in drinking water, was associated with prenatal lethality at the highest dose, with evidence

  11. Characterization of Network Structure of Polyacrylamide Based Hydrogels Prepared By Radiation Induced Polymerization

    SciTech Connect

    Mahmudi, Naim; Sen, Murat; Gueven, Olgun; Rendevski, Stojan

    2007-04-23

    In this study network structure of polyacrylamide based hydrogels prepared by radiation induced polymerization has been investigated. Polyacrylamide based hydrogels in the rod form were prepared by copolymerization of acrylamide(AAm) with hydroxyl ethyl methacrylate(HEMA) and methyl acrylamide(MAAm) in the presence of cross-linking agent and water by gamma rays at ambient temperature. Molecular weight between cross-links and effective cross-link density of hydrogels were calculated from swelling as well as shear modulus data obtained from compression tests. The results have shown that simple compression analyses can be used for the determination of effective cross-link density of hydrogels without any need to some polymer-solvent based parameters as in the case of swelling based determinations. Diffusion of water into hydrogels was examined by analyzing water absorption kinetics and the effect of network, structure on the diffusion type and coefficient was discussed.

  12. Synthesis of crosslinked starch-graft-polyacrylamide-co-sodium xanthate and its performances in wastewater treatment.

    PubMed

    Chang, Qing; Hao, Xuekui; Duan, Lili

    2008-11-30

    A novel crosslinked starch-graft-polyacrylamide-co-sodium xanthate (CSAX) was synthesized by grafting copolymerization reaction of corn starch, acrylamide (AM), and sodium xanthate using epichlorohydrin (EPI) as cross-linking reagent and ceric ammonium nitrate (CAN) as initiator in aqueous solution. CSAX was characterized by FTIR and element analysis. The performances of CSAX in wastewater treatment were evaluated by flocculation experiment. The results show that the CSAX was successfully synthesized and it has functions of removing both turbidity and copper ions from aqueous solution. It was proved that CSAX is more effective than crosslinked starch xanthate (CSX) and much more effective than crosslinked starch-graft-polyacrylamide (CSA) for removing copper ions. It was also proved that CSAX is little less effective than CSA, but much more effective than CSX for removing turbidity.

  13. Ink-native electrophoresis: an alternative to blue-native electrophoresis more suitable for in-gel detection of enzymatic activity.

    PubMed

    Kaneko, Keisuke; Sueyoshi, Noriyuki; Kameshita, Isamu; Ishida, Atsuhiko

    2013-09-15

    Blue-native electrophoresis (BNE) is a useful technique for analyzing protein complexes, but the Coomassie brilliant blue (CBB) dye used in BNE often hampers in-gel detection of enzymatic activity. Here we report an improved method, termed ink-native electrophoresis (INE), in which Pelikan 4001 fountain pen ink is used as a charge-shifting agent instead of CBB. INE is more suitable than BNE for in-gel detection of protein kinase activity after polyacrylamide gel electrophoresis (PAGE), and its performance in protein complex separation is comparable to that of conventional BNE. INE may provide a powerful tool to isolate and analyze various protein complexes.

  14. Effects of polyacrylamide soil conditioner on the iron status of soybean plants. [Glycine max

    SciTech Connect

    Wallace, A.; Wallace, G.A.; Abouzamzam, A.M.; Char, J.W.

    1986-05-01

    An iron-inefficient cultivar of soybean (Glycine max L. Merr. Bragg cv. PI-54619-5-1 was grown in two different calcareous soils, a Natrargid and a Torrifluvents, to determine if improvement of soil aeration with a synthetic polyacrylamide as a soil conditioner would decrease the tendency of the cultivar to lime-induced chlorosis. The results suggest that when soil is well aerated with good drainage from use of the soil conditioner, the iron status of plants is improved.

  15. EFFECT OF DEXTRAN-graft-POLYACRYLAMIDE INTERNAL STRUCTURE ON FLOCCULATION PROCESS PARAMETERS

    SciTech Connect

    Bezugla, T.; Kutsevol, N.; Shyichuk, A.; Ziolkowska, D.

    2008-08-28

    Dextran-graft-Polyacrylamide copolymers (D-g-PAA) of brush-like architecture were tested as flocculation aids in the model kaolin suspensions. Due to expanded conformation the D-g-PAA copolymers are more effective flocculants than individual PAA with close molecular mass. The internal structure of D-g-PAA copolymers which is determined by number and length of grafted PAA chains, the distance between grafts, etc., has the significant influence on flocculation behavior of such polymers.

  16. Fabricating polyacrylamide microbeads by inverse emulsification to mimic the size and elasticity of living cells.

    PubMed

    Labriola, Nicholas R; Mathiowitz, Edith; Darling, Eric M

    2016-12-20

    Inverse emulsification was used to fabricate polyacrylamide (PAAm) microbeads with size and elastic properties similar to typical, mammalian cells. These biomimicking microbeads could be fluorescently stained and functionalized with a collagen type-I coating, post-polymerization, for tracking bead locations and promoting cell recognition/binding, respectively. By occupying a previously unfilled range of sizes and mechanical properties, these microbeads may find unique use in both biomedical and materials applications.

  17. Agarose gel electrophoresis.

    PubMed

    Smith, D R

    1993-01-01

    After digestion of DNA with a restriction enzyme (Chapter 50), it is usually necessary, for both preparative and analytical purposes, to separate and visualize the products. In most cases, where the products are between 200 and 20,000 bp long, this is achieved by agarose gel electrophoresis. Agarose is a linear polymer that is extracted from seaweed and sold as a white powder. The powder is melted in buffer and allowed to cool, whereby the agarose forms a gel by hydrogen bonding. The hardened matrix contains pores, the size of which depends on the concentration of agarose. The concentration of agarose is referred to as a percentage of agarose to volume of buffer (w/v), and agarose gels are normally in the range of 0.3 to 3%. Many different apparatus arrangements have been devised to run agarose gels; for example, they can be run horizontally or vertically, and the current can be conducted by wicks or the buffer solution. However, today, the "submarine" gel system is almost universally used. In this method, the agarose gel is formed on a supporting plate, and then the plate is submerged into a tank containing a suitable electrophoresis buffer. Wells are preformed in the agarose gel with the aid of a "comb" that is inserted into the cooling agarose before the agarose has gelled. Into these wells are loaded the sample to be analyzed, which has been mixed with a dense solution (a loading buffer) to ensure that the sample sinks into the wells.

  18. A gel probe equilibrium sampler for measuring arsenic porewater profiles and sorption gradients in sediments: I. Laboratory development

    USGS Publications Warehouse

    Campbell, K.M.; Root, R.; O'Day, P. A.; Hering, J.G.

    2008-01-01

    A gel probe equilibrium sampler has been developed to study arsenic (As) geochemistry and sorption behavior in sediment porewater. The gels consist of a hydrated polyacrylamide polymer, which has a 92% water content. Two types of gels were used in this study. Undoped (clear) gels were used to measure concentrations of As and other elements in sediment porewater. The polyacrylamide gel was also doped with hydrous ferric oxide (HFO), an amorphous iron (Fe) oxyhydroxide. When deployed in the field, HFO-doped gels introduce a fresh sorbent into the subsurface thus allowing assessment of in situ sorption. In this study, clear and HFO-doped gels were tested under laboratory conditions to constrain the gel behavior prior to field deployment. Both types of gels were allowed to equilibrate with solutions of varying composition and re-equilibrated in acid for analysis. Clear gels accurately measured solution concentrations (??1%), and As was completely recovered from HFO-doped gels (??4%). Arsenic speciation was determined in clear gels through chromatographic separation of the re-equilibrated solution. For comparison to speciation in solution, mixtures of As(III) and As(V) adsorbed on HFO embedded in gel were measured in situ using X-ray absorption spectroscopy (XAS). Sorption densities for As(III) and As(V) on HFO embedded in gel were obtained from sorption isotherms at pH 7.1. When As and phosphate were simultaneously equilibrated (in up to 50-fold excess of As) with HFO-doped gels, phosphate inhibited As sorption by up to 85% and had a stronger inhibitory effect on As(V) than As(III). Natural organic matter (>200 ppm) decreased As adsorption by up to 50%, and had similar effects on As(V) and As(III). The laboratory results provide a basis for interpreting results obtained by deploying the gel probe in the field and elucidating the mechanisms controlling As partitioning between solid and dissolved phases in the environment. ?? 2008 American Chemical Society.

  19. Gel for Simultaneous Chemical Imaging of Anionic and Cationic Solutes Using Diffusive Gradients in Thin Films

    PubMed Central

    2013-01-01

    We report on a novel gel based on diffusive gradients in thin films (DGT) for the simultaneous measurement of cations and anions and its suitability for high resolution chemical imaging by using laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS). The new high resolution mixed binding gel (HR-MBG) is based on zirconium-hydroxide and suspended particulate reagent-iminodiacetate (SPR-IDA) as resin materials which are embedded in an ether-based urethane polymer hydrogel. The use of this polymer hydrogel material allows the production of ultrathin, highly stable and tear-proof resin gel layers with superior handling properties compared to existing ultrathin polyacrylamide gels. The gel was characterized regarding its uptake kinetics, the anion and cation capacities, and the effects of pH, ionic strength, and aging on the performance of the HR-MBG. Our results demonstrate the capability of this novel gel for concomitant sampling of anions and cations. The suitability of this new gel type for DGT chemical imaging at submm spatial resolution in soils using LA-ICPMS is shown. 2D images of P, As, Co, Cu, Mn, and Zn distributions around roots of Zea mays L. demonstrate the new opportunities offered by the HR-MBG for high-resolution mapping of solute dynamics in soil and sediment hotspots, such as the rhizosphere, by simultaneous observation of anionic and cationic solute species. PMID:24256092

  20. Microwave assisted synthesis of polyacrylamide grafted dextrin (Dxt-g-PAM): Development and application of a novel polymeric flocculant.

    PubMed

    Pal, Sagar; Nasim, T; Patra, A; Ghosh, S; Panda, A B

    2010-12-01

    An efficient polymeric flocculant was synthesized by microwave assisted grafting of polyacrylamide to dextrin. By varying the reaction conditions, various grades of graft copolymers were synthesized to obtain the optimized one. Viscometry, elemental analysis, FTIR spectroscopy, (13)C NMR spectroscopy, determination of molecular weight and radius of gyration using SLS analysis, thermal analysis and SEM analysis were employed to confirm that polyacrylamide has been grafted onto the dextrin backbone. The flocculation efficiency of the grafted products in kaolin suspension was dependent on the molecular weight, radius of gyration and length of the grafted polyacrylamide chains. The flocculant obtained by microwave assisted grafting method was superior to dextrin and polyacrylamide-based commercial flocculant (Rishfloc 226 LV) in flocculation tests.

  1. Development of a new aluminum/polymer gel system for permeability adjustment

    SciTech Connect

    Dovan, H.T.; Hutchins, R.D.

    1987-05-01

    A new method for gelling polyacrylamide with aluminum has been developed to reduce the effect of reservoir heterogeneity, resulting in improved waterflood efficiency and higher oil recovery. The method uses a soluble aluminum compound in a high-pH, nonreactive form that is mixed directly with the polymer at optimum concentrations. Polymer gelling occurs in the reservoir when reactive aluminum is generated by consumption of hydroxyl ions. Variations in gel strength and gel time are obtained by adjusting polymer and aluminate concentrations in the slug to the desired levels. This process has several advantages over the current aluminum citrate technology as well as the chromium redox bulk gel system. Although the process works best in freshwater systems, it can be modified to accommodate waters with higher brine content. Laboratory development of the process and a successful profile modification field trial are described.

  2. Band broadening in gel electrophoresis: scaling laws for the dispersion coefficient measured by FRAP.

    PubMed

    Tinland, B; Pernodet, N; Pluen, A

    1998-10-05

    We determined quantitatively the band broadening effect during gel electrophoresis by measuring the longitudinal dispersion coefficient Dx, with a fluorescence recovery after photobleaching setup, coupled to an electrophoretic cell. We carried out measurements as a function of the electric field, the average pore size, and the molecular length of DNA fragments. Our results are in good agreement with the predictions of the biased reptation model with fluctuations described by T. A. Duke et al. [(1992) Physics Review Letters, vol. 69, pp. 3260-3263]. This agreement is observed on single-stranded DNA [persistence length approximately equal to 4 nm; B. Tinland et al. (1997) Macromolecules, vol. 30, pp. 5763-5765] in polyacrylamide gels and on double-stranded DNA (persistence length approximately equal to 50 nm) in agarose gels, two systems where the ratio between the average pore size and the Kuhn length is larger than 1.

  3. Janus Gel Fabrication Using Liquid Drop Coalescence and Limited Mixing in the Hele-Shaw Geometry

    NASA Astrophysics Data System (ADS)

    Gonzalez, Brittany; Moran, Alexis; Lee, Donghee; Ryu, Sangjin

    2015-11-01

    Hydrogel substrates of tunable stiffness have been actively utilized for in vitro cell mechanobiology study. Here we present a new method to fabricate Janus polyacrylamide gel based on limited mixing between liquid drops coalescing in the Hele-Shaw geometry. Two pre-polymer drops with different concentrations were sandwiched and squeezed between two parallel glass surfaces. Once the drops coalesced in the decreased gap between the surfaces, gelation was initiated by UV light exposure with various time delays. AFM nano-indentation was utilized to map the Young's modulus of obtained gels. Fabricated Janus gels had two regions of different Young's moduli interfaced by the stiffness gradient zone, and the width of the gradient zone increased with the delay time. We acknowledge support from Bioengineering for Human Health grant from UNL and UNMC, and NSF REU grant for UNL.

  4. Effect of the exothermal polymerization reaction on polymer gel dosimetric measurements

    NASA Astrophysics Data System (ADS)

    Sedaghat, Mahbod; Bujold, Rachel; Lepage, Martin

    2010-11-01

    Discrepancies in polymer gel dosimetric measurements have been observed between containers of different sizes receiving the same radiation dose. We hypothesized that these deviations are caused by a change in the rate of polymerization due to internal heat increase in the gel containers resulting from the exothermic polymerization of monomers. Here, we test this hypothesis in a polyacrylamide gel dosimeter by recording the temperature in glass phantoms of different sizes during and after irradiation. The dose response of the samples was determined with magnetic resonance imaging. The difference of R2 values along the depth of the containers was below ±1%. We discuss that this small difference can be attributed to variations in the rate of gelatin cooling during manufacture rather than to the measured heat increase during irradiation.

  5. Conformance Improvement Using Gels

    SciTech Connect

    Seright, Randall S.; Schrader, Richard; II Hagstrom, John; Wang, Ying; Al-Dahfeeri, Abdullah; Gary, Raven; Marin; Amaury; Lindquist, Brent

    2002-09-26

    This research project had two objectives. The first objective was to identify gel compositions and conditions that substantially reduce flow through fractures that allow direct channeling between wells, while leaving secondary fractures open so that high fluid injection and production rates can be maintained. The second objective was to optimize treatments in fractured production wells, where the gel must reduce permeability to water much more than that to oil.

  6. Crystallization from Gels

    NASA Astrophysics Data System (ADS)

    Narayana Kalkura, S.; Natarajan, Subramanian

    Among the various crystallization techniques, crystallization in gels has found wide applications in the fields of biomineralization and macromolecular crystallization in addition to crystallizing materials having nonlinear optical, ferroelectric, ferromagnetic, and other properties. Furthermore, by using this method it is possible to grow single crystals with very high perfection that are difficult to grow by other techniques. The gel method of crystallization provides an ideal technique to study crystal deposition diseases, which could lead to better understanding of their etiology. This chapter focuses on crystallization in gels of compounds that are responsible for crystal deposition diseases. The introduction is followed by a description of the various gels used, the mechanism of gelling, and the fascinating phenomenon of Liesegang ring formation, along with various gel growth techniques. The importance and scope of study on crystal deposition diseases and the need for crystal growth experiments using gel media are stressed. The various crystal deposition diseases, viz. (1) urolithiasis, (2) gout or arthritis, (3) cholelithiasis and atherosclerosis, and (4) pancreatitis and details regarding the constituents of the crystal deposits responsible for the pathological mineralization are discussed. Brief accounts of the theories of the formation of urinary stones and gallstones and the role of trace elements in urinary stone formation are also given. The crystallization in gels of (1) the urinary stone constituents, viz. calcium oxalate, calcium phosphates, uric acid, cystine, etc., (2) the constituents of the gallstones, viz. cholesterol, calcium carbonate, etc., (3) the major constituent of the pancreatic calculi, viz., calcium carbonate, and (4) cholic acid, a steroidal hormone are presented. The effect of various organic and inorganic ions, trace elements, and extracts from cereals, herbs, and fruits on the crystallization of major urinary stone and gallstone

  7. Chirality Made Simple: A 1 - and 2-Dimensional Introduction to Stereochemistry

    ERIC Educational Resources Information Center

    Gawley, Robert E.

    2005-01-01

    The introduction of chirality in one and two dimensions, along with the concepts of internal and external reflection, can be combined with concepts familiar to all students. Once familiar with 1-Dimensional and 2-Dimensional chirality, the same concepts can be extended to 3-Dimensional and by projecting 3-D back to two, it is possible to interpret…

  8. Interactions of polyacrylamides used for enhanced oil recovery and reservoir isolates of the sulfate-reducing bacterium Desulfovibrio

    SciTech Connect

    Sewell, G.W.

    1987-01-01

    The interactions of partially hydrolyzed polyacrylamides utilized in enhanced oil recovery as mobility control agents and reservoir isolates of Desulfovibrio were examined. Produced waters from reservoirs undergoing polymer flooding were sampled to determine the presence and numbers of sulfate-reducing bacteria. The influence of polyacrylamide on the growth of Desulfovibrio under a number of conditions was studied. Brookfield viscosity and screen factor measurements were used to screen for adverse changes in the rheological properties of polyacrylamide solutions which had been exposed to Desulfovibrio. Partially hydrolyzed polyacrylamides stimulated the growth of Desulfovibrio under laboratory conditions. The polymer may act as a nitrogen source or as an incomplete substrate under certain conditions, but is apparently not utilizable as a carbon source. Desulfovibrio caused a loss in screen factor of polyacrylamide solutions under anaerobic conditions, but no significant loss in solution viscosity was observed. No decrease in polymer concentration was detected. Labelled polyacrylamide was not incorporated in Desulfovibrio. The authors conclude that the observed screen factor losses are probably not the result of enzymatic degradation of the polymer, and are most likely a physical/chemical interaction of polymer molecules with some product of Desulfovibrio metabolism.

  9. Top-down, bottom-up, and side-to-side proteomics with virtual 2-D gels

    NASA Astrophysics Data System (ADS)

    Ogorzalek Loo, Rachel R.; Hayes, Richard; Yang, Yanan; Hung, Frank; Ramachandran, Prasanna; Kim, Nuri; Gunsalus, Robert; Loo, Joseph A.

    2005-02-01

    Intact protein masses can be measured directly from immobilized pH gradient (IPG) isoelectric focusing (IEF) gels loaded with mammalian and prokaryotic samples, as demonstrated here with murine macrophage and Methanosarcina acetivorans cell lysates. Mass accuracy and resolution is improved by employing instruments which decouple the desorption event from mass measurement; e.g., quadrupole time-of-flight instruments. MALDI in-source dissociation (ISD) is discussed as a means to pursue top-down sequencing for protein identification. Methods have been developed to enzymatically digest all proteins in an IEF gel simultaneously, leaving the polyacrylamide gel attached to its polyester support. By retaining all gel pieces and their placement relative to one another, sample handling and tracking are minimized, and comparison to 2-D gel images is facilitated. MALDI-MS and MS/MS can then be performed directly from dried, matrix-treated IPG strips following whole-gel trypsin digestion, bottom-up methodology. Side-to-side proteomics, highlighting the link between virtual and classical 2-D gel electrophoresis, is introduced to describe a method whereby intact masses are measured from one side (the IEF gel), while proteins are identified based on analyses performed from the other side (the SDS-PAGE gel).

  10. Cu2+-assisted two dimensional charge-mass double focusing gel electrophoresis and mass spectrometric analysis of histone variants.

    PubMed

    Zhang, Wenyang; Tang, Xuemei; Ding, Mengjie; Zhong, Hongying

    2014-12-10

    Abundant isoforms and dynamic posttranslational modifications cause the separation and identification of histone variants to be experimentally challenging. To meet this need, we employ two-dimensional electrophoretic gel separation followed by mass spectrometric detection which takes advantage of the chelation of Cu(2+) with amino acid residues exposed on the surfaces of the histone proteins. Acid-extracted rat liver histones were first mixed with CuSO4 solution and then separated in one dimension with triton-acid-urea (TAU) gel electrophoresis and in a second dimension using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The separations result from both the changes in charge and mass upon Cu(2+) chelation. Identities of each separated gel bands were obtained by using matrix-assisted laser desorption-ionization mass spectrometry (MALDI-MS). It was found that the migration of H3 histone isoforms of rat liver is markedly affected by the use of Cu(2+) ions.

  11. CONFORMANCE IMPROVEMENT USING GELS

    SciTech Connect

    Randall S. Seright

    2002-02-28

    This technical progress report describes work performed from June 20 through December 19, 2001, for the project, ''Conformance Improvement Using Gels''. Interest has increased in some new polymeric products that purport to substantially reduce permeability to water while causing minimum permeability reduction to oil. In view of this interest, we are currently studying BJ's Aqua Con. Results from six corefloods revealed that the Aqua Con gelant consistently reduced permeability to water more than that to oil. However, the magnitude of the disproportionate permeability reduction varied significantly for the various experiments. Thus, as with most materials tested to date, the issue of reproducibility and control of the disproportionate permeability remains to be resolved. Concern exists about the ability of gels to resist washout after placement in fractures. We examined whether a width constriction in the middle of a fracture would cause different gel washout behavior upstream versus downstream of the constriction. Tests were performed using a formed Cr(III)-acetate-HPAM gel in a 48-in.-long fracture with three sections of equal length, but with widths of 0.08-, 0.02-, and 0.08-in., respectively. The pressure gradients during gel extrusion (i.e., placement) were similar in the two 0.08-in.-wide fracture sections, even though they were separated by a 0.02-in.-wide fracture section. The constriction associated with the middle fracture section may have inhibited gel washout during the first pulse of brine injection after gel placement. However, during subsequent phases of brine injection, the constriction did not inhibit washout in the upstream fracture section any more than in the downstream section.

  12. Two-dimensional gel electrophoresis in bacterial proteomics.

    PubMed

    Curreem, Shirly O T; Watt, Rory M; Lau, Susanna K P; Woo, Patrick C Y

    2012-05-01

    Two-dimensional gel electrophoresis (2-DE) is a gel-based technique widely used for analyzing the protein composition of biological samples. It is capable of resolving complex mixtures containing more than a thousand protein components into individual protein spots through the coupling of two orthogonal biophysical separation techniques: isoelectric focusing (first dimension) and polyacrylamide gel electrophoresis (second dimension). 2-DE is ideally suited for analyzing the entire expressed protein complement of a bacterial cell: its proteome. Its relative simplicity and good reproducibility have led to 2-DE being widely used for exploring proteomics within a wide range of environmental and medically-relevant bacteria. Here we give a broad overview of the basic principles and historical development of gel-based proteomics, and how this powerful approach can be applied for studying bacterial biology and physiology. We highlight specific 2-DE applications that can be used to analyze when, where and how much proteins are expressed. The links between proteomics, genomics and mass spectrometry are discussed. We explore how proteomics involving tandem mass spectrometry can be used to analyze (post-translational) protein modifications or to identify proteins of unknown origin by de novo peptide sequencing. The use of proteome fractionation techniques and non-gel-based proteomic approaches are also discussed. We highlight how the analysis of proteins secreted by bacterial cells (secretomes or exoproteomes) can be used to study infection processes or the immune response. This review is aimed at non-specialists who wish to gain a concise, comprehensive and contemporary overview of the nature and applications of bacterial proteomics.

  13. Polyacrylamide-hydroxyapatite composite: Preparation, characterization and adsorptive features for uranium and thorium

    SciTech Connect

    Baybas, Demet; Ulusoy, Ulvi

    2012-10-15

    The composite of synthetically produced hydroxyapatite (HAP) and polyacrylamide was prepared (PAAm-HAP) and characterized by BET, FT-IR, TGA, XRD, SEM and PZC analysis. The adsorptive features of HAP and PAAm-HAP were compared for UO{sub 2}{sup 2+} and Th{sup 4+}. The entrapment of HAP into PAAm-HAP did not change the structure of HAP. Both structures had high affinity to the studied ions. The adsorption capacity of PAAm-HAP was than that of HAP. The adsorption dependence on pH and ionic intensity provided supportive evidences for the effect of complex formation on adsorption process. The adsorption kinetics was well compatible to pseudo second order model. The values of enthalpy and entropy changes were positive. Th{sup 4+} adsorption from the leachate obtained from a regional fluorite rock confirmed the selectivity of PAAm-HAP for this ion. In consequence, PAAm-HAP should be considered amongst favorite adsorbents for especially deposition of nuclear waste containing U and Th, and radionuclide at secular equilibrium with these elements. - Graphical abstract: SEM images of hydroxyapatite (HAP) and polyacrylamide-hydroxyapatite (PAAm-HAP), and the adsorption isotherms for Uranium and Thorium. Highlights: Black-Right-Pointing-Pointer Composite of PAAm-HAP was synthesized from hydroxyapatite and polyacrylamide. Black-Right-Pointing-Pointer The materials were characterized by BET, FT-IR, XRD, SEM, TGA and PZC analysis. Black-Right-Pointing-Pointer HAP and PAAm-HAP had high sorption capacity and very rapid uptake for UO{sub 2}{sup 2+} and Th{sup 4+}. Black-Right-Pointing-Pointer Super porous PAAm was obtained from PAAm-HAP after its removal of HAP content. Black-Right-Pointing-Pointer The composite is potential for deposition of U, Th and its associate radionuclides.

  14. Templateless synthesis of polyacrylamide-based Nanogels via RAFT dispersion polymerization.

    PubMed

    Ma, Kai; Xu, Yuanyuan; An, Zesheng

    2015-03-01

    This paper reports on the synthesis of well-defined polyacrylamide-based nanogels via reversible addition-fragmentation chain transfer (RAFT) dispersion polymerization, highlighting a templateless route for the efficient synthesis of nanogels based on water-soluble polymers. RAFT dispersion polymerization of acrylamide in co-nonsolvents of water-tert-butanol mixtures by chain extension from poly(dimethylacrylamide) shows well-controlled polymerization process, uniform nanogel size, and excellent colloidal stability. The versatility of this approach is further demonstrated by introducing a hydrophobic co-monomer (butyl acrylate) without disturbing the dispersion polymerization process.

  15. Irradiation mediated synthesis of a superabsorbent hydrogel network based on polyacrylamide grafted onto salep

    NASA Astrophysics Data System (ADS)

    Bardajee, Ghasem Rezanejade; Pourjavadi, Ali; Soleyman, Rouhollah; Sheikh, Nasrin

    2008-09-01

    The synthesis and swelling behavior of a new superabsorbent hydrogel based on natural salep grafted with polyacrylamide is described. The new biopolymer was synthesized via simultaneous crosslinking and graft copolymerization of acrylamide monomer onto a salep backbone using radiochemical methods. Various parameters such as relative contents of salep and acrylamide, as well as total dose of γ-rays were examined. The best synthesis condition is reported and a mechanism for superabsorbent hydrogel formation suggested. Factors affecting the swelling behavior of hydrogel were also studied.

  16. Partially imidized, water-soluble polymeric amides. I. Partially imidized polyacrylamide and polymethacrylamide

    SciTech Connect

    Shepitka, J.S.; Case, C.E.; Donaruma, L.G.; Hatch, M.J.; Kilmer, N.H.; Khune, G.D.; Martin, F.D.; Ward, J.S.; Wilson, K.V.

    1983-12-01

    Five to six million molecular weight polyacrylamide and polymethacrylamides of comparable post-alkaline hydrolysis viscosities were imidized by dissolution and heating in 6N HCl. After alkaline hydrolysis, the imidized polymers demonstrated significantly better retention of viscosity to 2% NaCl than did similar partially hydrolyzed polymers. Viscosities in 0.01% NaCl and resistance to shear were not markedly affected by this modification. It is assumed that this improved performance in brine is the result of chain stiffening due to intrachain imide rings. 2 figures, 2 tables.

  17. The roles of disulphide and non-covalent bonding in the functional properties of heat-induced whey protein gels.

    PubMed

    Havea, Palatasa; Carr, Alistair J; Creamer, Lawrence K

    2004-08-01

    Heat-induced gelation (80 degrees C, 30 min or 85 degrees C, 60 min) of whey protein concentrate (WPC) solutions was studied using transmission electron microscopy (TEM), dynamic rheology and polyacrylamide gel electrophoresis (PAGE). The WPC solutions (150 g/kg, pH 6.9) were prepared by dispersing WPC powder in water (control), 10 g/kg sodium dodecyl sulphate (SDS) solution or 10 mM-dithiothreitol (DTT) solution. The WPC gels containing SDS were more translucent than the control gels, which were slightly more translucent than the gels containing DTT. TEM analyses showed that the SDS-gels had finer aggregate structure (approximately equal to 10 nm) than the control gels (approximately equal to 100 nm), whereas the DTT-gels had a more particulate structure (approximately equal to 200 to 300 nm). Dynamic rheology measurements showed that the control WPC gels had storage modulus (G) values (approximately equal to 13,500 Pa) that were approximately equal to 25 times higher than those of the SDS-gels (approximately equal to 550 Pa) and less than half those of the DTT-gels after cooling. Compression tests showed that the DTT-gels were more rigid and more brittle than the control gels, whereas the SDS-gels were softer and more rubbery than either the control gels or the DTT-gels. PAGE analyses of WPC gel samples revealed that the control WPC solutions heated at 85 degrees C for 10 min contained both disulphide bonds and non-covalent linkages. In both the SDS-solutions and the DTT-solutions, the denatured whey protein molecules were in the form of monomers or small aggregates. It is likely that, on more extended heating, more disulphide linkages were formed in the SDS-gels whereas more hydrophobic aggregates were formed in the DTT-gels. These results demonstrate that the properties of heat-induced WPC gels are strongly influenced by non-covalent bonding. Intermolecular disulphide bonds appeared to give the rubbery nature of heat-induced WPC gels whereas non-covalent bonds

  18. Spatially resolved multicomponent gels

    NASA Astrophysics Data System (ADS)

    Draper, Emily R.; Eden, Edward G. B.; McDonald, Tom O.; Adams, Dave J.

    2015-10-01

    Multicomponent supramolecular systems could be used to prepare exciting new functional materials, but it is often challenging to control the assembly across multiple length scales. Here we report a simple approach to forming patterned, spatially resolved multicomponent supramolecular hydrogels. A multicomponent gel is first formed from two low-molecular-weight gelators and consists of two types of fibre, each formed by only one gelator. One type of fibre in this ‘self-sorted network’ is then removed selectively by a light-triggered gel-to-sol transition. We show that the remaining network has the same mechanical properties as it would have done if it initially formed alone. The selective irradiation of sections of the gel through a mask leads to the formation of patterned multicomponent networks, in which either one or two networks can be present at a particular position with a high degree of spatial control.

  19. Patterns in shrinking gels

    NASA Astrophysics Data System (ADS)

    Matsuo, Eriko Sato; Tanaka, Toyoichi

    1992-08-01

    POLYMER gels can undergo a volume phase transition (either continuous or discontinuous) when an external condition, such as temperature or solvent composition, is altered1-3. During this transition, the volume may change by a factor of several thousand, and various patterns develop in the gel. The patterns arising from swelling and shrinking differ in both their appearance and their physical mechanisms. The mechanism for the formation and evolution of patterns on swelling gels has been established as being due to a single kind of mechanical instability4-7 in contrast, the shrinking patterns seem to be sensitive to both the initial and final states of the transition. Here we classify the various shrinking patterns in the form of a phase diagram, and explain the poly-morphism in terms of macroscopic phase separation.

  20. The dose response of normoxic polymer gel dosimeters measured using X-ray CT.

    PubMed

    Hill, B; Venning, A; Baldock, C

    2005-07-01

    X-ray CT was used to determine the dose response of normoxic polymer gel dosimeters. Normoxic polymer gel dosimeters were manufactured and irradiated up to 150 Gy. Up to 50 CT images were acquired on a Toshiba Aquilion Multislice CT scanner using protocols for 80 kV and 135 kV to determine dose response. HU-dose sensitivity, the linear regression of data for the HU versus dose for the linear part of the curve up to 60 Gy was 0.38+/-0.07 HU Gy(-1) for 135 kV and 0.37+/-0.01 HU Gy(-1) for 80 kV. Dose resolution was found to be < 1.3 Gy for an absorbed dose range up to 70 Gy for 135 kV, similar to that measured previously for polyacrylamide gel (PAG). Although the HU-dose sensitivity was lower than that previously measured for PAG gel dosimeters it had a greater range of absorbed dose indicating that normoxic polymer gel dosimeters have potential in CT gel dosimetry.

  1. Interfacing solid-state nanopores with gel media to slow DNA translocations.

    PubMed

    Waugh, Matthew; Carlsen, Autumn; Sean, David; Slater, Gary W; Briggs, Kyle; Kwok, Harold; Tabard-Cossa, Vincent

    2015-08-01

    We demonstrate the ability to slow DNA translocations through solid-state nanopores by interfacing the trans side of the membrane with gel media. In this work, we focus on two reptation regimes: when the DNA molecule is flexible on the length scale of a gel pore, and when the DNA behaves as persistent segments in tight gel pores. The first regime is investigated using agarose gels, which produce a very wide distribution of translocation times for 5 kbp dsDNA fragments, spanning over three orders of magnitude. The second regime is attained with polyacrylamide gels, which can maintain a tight spread and produce a shift in the distribution of the translocation times by an order of magnitude for 100 bp dsDNA fragments, if intermolecular crowding on the trans side is avoided. While previous approaches have proven successful at slowing DNA passage, they have generally been detrimental to the S/N, capture rate, or experimental simplicity. These results establish that by controlling the regime of DNA movement exiting a nanopore interfaced with a gel medium, it is possible to address the issue of rapid biomolecule translocations through nanopores-presently one of the largest hurdles facing nanopore-based analysis-without affecting the signal quality or capture efficiency.

  2. Dialect Distance Assessment Based on 2-Dimensional Pitch Slope Features and Kullback Leibler Divergence

    DTIC Science & Technology

    2009-04-08

    DATES COVERED (From - To) March 2008 – April 2009 4. TITLE AND SUBTITLE DIALECT DISTANCE ASSESSMENT BASED ON 2-DIMENSIONAL PITCH SLOPE FEATURES AND...rights are reserved by the copyright owner. 14. ABSTRACT Dialect variations of a language have a severe impact on the performance of speech systems...Knowing how close or separate dialects are in a given language space provides useful information to predict or improve, system performance when

  3. Large-Eddy Simulation on turbulent flow and plume dispersion over a 2-dimensional hill

    NASA Astrophysics Data System (ADS)

    Nakayama, H.; Nagai, H.

    2010-05-01

    The dispersion analysis of airborne contaminants including radioactive substances from industrial or nuclear facilities is an important issue for air quality maintenance and safety assessment. In Japan, many nuclear power plants are located at complex coastal terrains. In these cases, terrain effects on the turbulent flow and plume dispersion should be investigated. In this study, we perform Large-Eddy Simulation (LES) of turbulent flow and plume dispersion over a 2-dimensional hill flow and investigate the characteristics of mean and fluctuating concentrations.

  4. Gravitational compression of colloidal gels

    NASA Astrophysics Data System (ADS)

    Liétor-Santos, J. J.; Kim, C.; Lu, P. J.; Fernández-Nieves, A.; Weitz, D. A.

    2009-02-01

    We study the compression of depletion gels under the influence of a gravitational stress by monitoring the time evolution of the gel interface and the local volume fraction, φ , inside the gel. We find φ is not constant throughout the gel. Instead, there is a volume fraction gradient that develops and grows along the gel height as the compression process proceeds. Our results are correctly described by a non-linear poroelastic model that explicitly incorporates the φ -dependence of the gravitational, elastic and viscous stresses acting on the gel.

  5. Investigation on efficient adsorption of cationic dyes on porous magnetic polyacrylamide microspheres.

    PubMed

    Yao, Tong; Guo, Song; Zeng, Changfeng; Wang, Chongqing; Zhang, Lixiong

    2015-07-15

    We report here the preparation of porous magnetic polyacrylamide microspheres for efficient removal of cationic dyes by a simple polymerization-induced phase separation method. Characterizations by various techniques indicate that the microspheres show porous structures and magnetic properties. They can adsorb methylene blue with high efficiency, with adsorption capacity increasing from 263 to 1977 mg/g as the initial concentration increases from 5 to 300 mg/L. Complete removal of methylene blue can be obtained even at very low concentrations. The equilibrium data is well described by the Langmuir isotherm models, exhibiting a maximum adsorption capacity of 1990 mg/g. The adsorption capacity increases with increasing initial pH and reaches a maximum at pH 8, revealing an electrostatic interaction between the microspheres and the methylene blue molecules. The microspheres also show high adsorption capacities for neutral red and gentian violet of 1937 and 1850 mg/g, respectively, as well as high efficiency in adsorption of mixed-dye solutions. The dye-adsorbed magnetic polyacrylamide microspheres can be easily desorbed, and can be repeatedly used for at least 6 cycles without losing the adsorption capacity. The adsorption capacity and efficiency of the microspheres are much higher than those of reported adsorbents, which exhibits potential practical application in removing cationic dyes.

  6. Acrylamide and polyacrylamide: a review of production, use, environmental fate and neurotoxicity.

    PubMed

    Smith, E A; Oehme, F W

    1991-01-01

    Acrylamide is a highly water soluble vinyl monomer formed from the hydration of acrylonitrile. The major commercial use of acrylamide is the formation of polymers. In the environment acrylamide has a high mobility in soil, may travel great distances in ground-water, is biodegradable, and is not absorbed by sediments or affected by water treatment. It is absorbed by all routes of animal exposure. The main metabolite is N-acetyl-S-(3-amino-3-oxypropyl)-cysteine and is excreted predominantly in the urine. Acrylamide produces an ascending central/peripheral axonopathy in man and animals. The major histological findings are swelling of axons and/or decrease in number of large diameter axons. Acrylamide axonopathy is reversible with time, but full recovery depends upon the severity of the intoxication. All reported cases of acrylamide toxicity have been attributed to handling the monomer. Polyacrylamide is non-toxic. Specific clinical features of acrylamide intoxication are more conclusive than electrophysiological, histological or biochemical laboratory tests for diagnosis. Acrylamide can be detected by titration, colorimetry, high performance chromatography, gas chromatography and polarography in air, water, biological fluids, tissues and polyacrylamides. Present research on the effects of acrylamide focuses on developmental and reproductive effects, genotoxicity and carcinogenicity.

  7. Rheology of Active Gels

    NASA Astrophysics Data System (ADS)

    Chen, Daniel

    2015-03-01

    Active networks drive a diverse range of critical processes ranging from motility to division in living cells, yet a full picture of their rheological capabilities in non-cellular contexts is still emerging, e.g., How does the rheological response of a network capable of remodeling under internally-generated stresses differ from that of a passive biopolymer network? In order to address this and other basic questions, we have engineered an active gel composed of microtubules, bidirectional kinesin motors, and molecular depletant that self-organizes into a highly dynamic network of active bundles. The network continually remodels itself under ATP-tunable cycles of extension, buckling, fracturing, and self-healing. Using confocal rheometry we have simultaneously characterized the network's linear and non-linear rheological responses to shear deformation along with its dynamic morphology. We find several surprising and unique material properties for these active gels; most notably, rheological cloaking, the ability of the active gel to drive large-scale fluid mixing over several orders of flow magnitude while maintaining an invariant, solid-like rheological profile and spontaneous flow under confinement, the ability to exert micro-Newton forces to drive persistent directed motion of the rheometer tool. Taken together, these results and others to be discussed highlight the rich stress-structure-dynamics relationships in this class of biologically-derived active gels.

  8. Enzymatic assessment of cholesterol on electrophoresis gels for estimating HDL size distribution and plasma concentrations of HDL subclasses[S

    PubMed Central

    Toledo-Ibelles, Paola; García-Sánchez, Cynthia; Ávila-Vazzini, Nydia; Carreón-Torres, Elizabeth; Posadas-Romero, Carlos; Vargas-Alarcón, Gilberto; Pérez-Méndez, Oscar

    2010-01-01

    The aim of this study was to develop an enzymatic cholesterol staining method to determine HDL subclasses in a polyacrylamide gradient gel electrophoresis, which further allows staining by protein in the same electrophoresis lane. HDLs from 120 healthy individuals were separated through nondenaturing PAGE. HDLs were stained for cholesterol using an enzymatic semisolid mixture. Once the gels were unstained, they were stained again for proteins with Coomassie blue. The proportions of HDL subclasses were determined by densitometry. HDL subclasses were transformed to concentrations using as reference HDL-cholesterol plasma levels. This method is comparable in linearity and reproducibility to Coomassie blue staining, although it provides quantitative data. As expected, HDL size distribution shifted toward larger particles when determined by cholesterol as compared with protein. With this method, we observed different proportions of HDL subclasses between men and women as compared with Coomassie blue staining. We described a method to determine HDL size distribution by enzymatic cholesterol staining on polyacrylamide gels. The method allows the quantification of the cholesterol plasma concentration of each HDL subclass with the possibility to further stain the protein in the same sample. The combination of HDL staining by cholesterol and protein on electrophoresis gels provides information that may have clinical relevance. PMID:20097938

  9. Large-Eddy Simulation on Turbulent Flow and Plume Dispersion over A 2-Dimensional Hill

    NASA Astrophysics Data System (ADS)

    Nakayama, H.; Nagai, H.

    2009-09-01

    The dispersion analysis of airborne contaminant including radioactive substances from industrial or nuclear facilities is the important issue for maintenance of air quality or safety assessment. Many studies on the plume dispersion behavior in the simulated atmospheric boundary layer flow over flat plain have been conducted mainly by wind tunnel experiments. However, many nuclear power plants are located at complex coastal terrains in Japan. In this case, topographical effects on the turbulent flow and plume dispersion should be investigated. Therefore, we perform Large-Eddy Simulations (LES) on turbulent flows and plume dispersions. As the first step of this study, we try it for a 2-dimensional hill flow and investigate the characteristics of mean and fluctuation concentrations. In order to produce a realistic turbulent boundary layer flow, we set up two computational domains. One is the driver region for generating the spatially-developing boundary layer flow and the other is the main computational region for plume dispersion over a 2-dimensional hill. Near the inlet of the driver region, roughness blocks for generating a thick turbulent boundary layer are placed. The inflow turbulence data obtained in the driver region are imposed at the inlet of the main computational domain at each time step. The sizes of driver and main computational regions are 46.25H×6.25H×25.0H and 37.5H×6.25H×25.0H (H: the height of a 2-dimensional hill) in x-, y- and z-directions, respectively. Here, x, y and z indicate streamwise, spanwise and vertical directions, respectively. The number of grid points of driver and main computational regions are 400×100×90 and 400×100×90 in x, y and z directions, respectively. A 2-dimensional hill model has 5.45H length and are placed at a distance of 10.4H downstream from the inlet of the main computational region. A release point of a tracer gas is located at a distance of 9.09H upstream from a 2-dimensional hill top and elevated with the

  10. The use of time-resolved fluorescence in gel-based proteomics for improved biomarker discovery

    NASA Astrophysics Data System (ADS)

    Sandberg, AnnSofi; Buschmann, Volker; Kapusta, Peter; Erdmann, Rainer; Wheelock, Åsa M.

    2010-02-01

    This paper describes a new platform for quantitative intact proteomics, entitled Cumulative Time-resolved Emission 2-Dimensional Gel Electrophoresis (CuTEDGE). The CuTEDGE technology utilizes differences in fluorescent lifetimes to subtract the confounding background fluorescence during in-gel detection and quantification of proteins, resulting in a drastic improvement in both sensitivity and dynamic range compared to existing technology. The platform is primarily designed for image acquisition in 2-dimensional gel electrophoresis (2-DE), but is also applicable to 1-dimensional gel electrophoresis (1-DE), and proteins electroblotted to membranes. In a set of proof-of-principle measurements, we have evaluated the performance of the novel technology using the MicroTime 100 instrument (PicoQuant GmbH) in conjunction with the CyDye minimal labeling fluorochromes (GE Healthcare, Uppsala, Sweden) to perform differential gel electrophoresis (DIGE) analyses. The results indicate that the CuTEDGE technology provides an improvement in the dynamic range and sensitivity of detection of 3 orders of magnitude as compared to current state-of-the-art image acquisition instrumentation available for 2-DE (Typhoon 9410, GE Healthcare). Given the potential dynamic range of 7-8 orders of magnitude and sensitivities in the attomol range, the described invention represents a technological leap in detection of low abundance cellular proteins, which is desperately needed in the field of biomarker discovery.

  11. Characterization of nanocellulose reinforced semi-interpenetrating polymer network of poly(vinyl alcohol) & polyacrylamide composite films.

    PubMed

    Mandal, Arup; Chakrabarty, Debabrata

    2015-12-10

    Semi-interpenetrating polymer network (semi-IPN) of poly(vinyl alcohol)/polyacrylamide was reinforced with various doses of nanocellulose. The different composite films thus prepared were characterized with respect to their mechanical, thermal, morphological and barrier properties. The composite film containing 5 wt.% of nanocellulose showed the highest tensile strength. The semi-interpenetrating polymer network of poly(vinyl alcohol)/polyacrylamide; and its various composites with nanocellulose were almost identical in their thermal stability. Each of the composites however exhibited much superior stability with respect to the linear poly(vinyl alcohol) and crosslinked polyacrylamide. The scanning electron microscopy (SEM) and atomic force microscopy (AFM) studies exhibited phase separated morphology where agglomerates of nanocellulose were found to be dispersed in the matrix of the semi-IPN. The moisture vapor transmission rate (MVTR) was the lowest for the film containing 5 wt.% of nanocellulose.

  12. Self-organization and oscillation of negatively charged dust particles in a 2-dimensional dusty plasma

    NASA Astrophysics Data System (ADS)

    Song, Y. L.; Huang, F.; Chen, Z. Y.; Liu, Y. H.; Yu, M. Y.

    2016-02-01

    Negatively charged dust particles immersed in 2-dimensional dusty plasma system are investigated by molecular dynamics simulations. The effects of the confinement potential and attraction interaction potential on dust particle self-organization are studied in detail and two typical dust particle distributions are obtained when the system reaches equilibrium. The average radial velocity (ARV), average radial force (ARF) and radial mean square displacement are employed to analyze the dust particles' dynamics. Both ARVs and ARFs exhibit oscillation behaviors when the simulation system reaches equilibrium state. The relationships between the oscillation and confinement potential and attraction potential are studied in this paper. The simulation results are qualitatively similar to experimental results.

  13. Hydrodynamic-type systems describing 2-dimensional polynomially integrable geodesic flows

    NASA Astrophysics Data System (ADS)

    Manno, Gianni; Pavlov, Maxim V.

    2017-03-01

    Starting from a homogeneous polynomial in momenta of arbitrary order we extract multi-component hydrodynamic-type systems which describe 2-dimensional geodesic flows admitting the initial polynomial as integral. All these hydrodynamic-type systems are semi-Hamiltonian, thus implying that they are integrable according to the generalized hodograph method. Moreover, they are integrable in a constructive sense as polynomial first integrals allow to construct generating equations of conservation laws. According to the multiplicity of the roots of the polynomial integral, we separate integrable particular cases.

  14. Dynamical analysis and simulation of a 2-dimensional disease model with convex incidence

    NASA Astrophysics Data System (ADS)

    Yu, Pei; Zhang, Wenjing; Wahl, Lindi M.

    2016-08-01

    In this paper, a previously developed 2-dimensional disease model is studied, which can be used for both epidemiologic modeling and in-host disease modeling. The main attention of this paper is focused on various dynamical behaviors of the system, including Hopf and generalized Hopf bifurcations which yield bistability and tristability, Bogdanov-Takens bifurcation, and homoclinic bifurcation. It is shown that the Bogdanov-Takens bifurcation and homoclinic bifurcation provide a new mechanism for generating disease recurrence, that is, cycles of remission and relapse such as the viral blips observed in HIV infection.

  15. Heat-induced whey protein gels: protein-protein interactions and functional properties.

    PubMed

    Havea, Palatasa; Watkinson, Philip; Kuhn-Sherlock, Barbara

    2009-02-25

    Heat-induced gelation (80 degrees C for 30 min or 85 degrees C for 60 min) of whey protein concentrate (WPC) solutions was studied using small deformation dynamic rheology, small and large deformation compression, and polyacrylamide gel electrophoresis (PAGE). The WPC solutions (15% w/w, pH 6.9) were prepared by dispersing WPC powder in water (control), 1% (w/w) sodium dodecyl sulfate (SDS) solution, and N-ethylmaleimide (NEM) solution at a protein/NEM molar ratio of 1:1 or in 10 mM dithiothreitol (DTT) solution. PAGE analyses showed that the heat treatment of control solutions contained both disulfide and non-covalent linkages between denatured protein molecules. Only disulfide linkages were formed in heated SDS-WPC solutions, whereas only non-covalent linkages were formed in DTT-WPC and NEM-WPC solutions during heating. In heated NEM-WPC solutions, the pre-existing disulfide linkages remained unaltered. Small deformation rheology measurements showed that the storage modulus (G') values, compared with those of the control WPC gels (approximately 14000 Pa), were 3 times less for the SDS-WPC gels (approximately 4000 Pa), double for the NEM-WPC gels (approximately 24000 Pa), and even higher for the DTT-WPC gels (approximately 30000 Pa). Compression tests suggested that the rubberiness (fracture strain) of the WPC gels increased as the degree of disulfide linkages within the gels increased, whereas the stiffness (modulus) of the gels increased as the degree of non-covalent associations among the denatured protein molecules increased.

  16. Transient association of the DNA-ligand complex during gel electrophoresis.

    PubMed

    Protozanova, E; Macgregor, R B

    1999-07-01

    DNA frayed wires are extremely stable multistranded complexes arising from the association of oligonucleotides with long terminal runs of consecutive guanines. Frayed wires originating from d(A15G15) have multiple binding sites for short complementary oligonucleotides such as dT10. We examine unusual band patterns obtained when complexes formed between dT10 and DNA frayed wires are resolved on nondenaturing polyacrylamide gels. Since the lifetime of the dT10-frayed wire complexes is shorter than the time of the gel run, the interaction between the components during the gel electrophoresis affects their band patterns. We have conducted chasing experiments to show that (i) the binding of dT10 to the frayed wires can occur during gel electrophoresis, and (ii) dissociation of the complexes occurs during the gel run. Rapid repetitive dissociation-reassociation of the complexes leads to a constant partitioning of dT10 between their binding sites within frayed wires. Consequently, complexes composed of frayed wires and various numbers of bound ligands appear on the gel as a single well-defined band. The mobilities of these bands decrease continuously with the concentration of the ligand reaching saturation when all the binding sites are occupied. This characteristic pattern is observed only for relatively unstable interactions. Longer ligands, i.e., oligonucleotides with higher affinity towards the binding sites, cease to exhibit the dynamic character of interaction during gel electrophoresis. These ligands form long-lived complexes with the frayed wires that appear on the gel as faint smeared bands reflecting the presence of multiple stable complexes.

  17. Coupling isoelectric focusing gel electrophoresis to mass spectrometry by electrostatic spray ionization.

    PubMed

    Qiao, Liang; Tobolkina, Elena; Liu, Baohong; Girault, Hubert H

    2013-05-07

    Gel electrophoresis has been used for decades as a high-resolution separation technique for proteins and protein isomers but has been limited in the coupling with MS because of low throughput and poor automaticity compared with LC-MS. In this work, we have developed an ambient ionization strategy, electrostatic spray ionization, for in situ ionization of proteins or peptides inside a surfactant-free polyacrylamide gel. The samples can be first separated by isoelectric focusing in a gel and then quickly in situ detected by scanning the gel with the electrostatic spray ionization mass spectrometry. With this strategy, nanograms of proteins or peptides inside a band are enough to be ionized for MS detection. This method for protein/peptide spots visualization is sensitive, providing sample molecular weight information while avoiding spot staining and chemical extraction procedures that can introduce contaminants and sample loss. Proof-of-principle results have demonstrated that the electrostatic spray ionization can produce sample ions from a complex background, and with a spatial resolution matching the isoelectric focusing, it is therefore a good choice to couple directly isoelectric focusing gel electrophoresis with mass spectrometry.

  18. Development of a gel formulation of formic acid for control of parasitic mites of honey bees.

    PubMed

    Kochansky, J; Shimanuki, H

    1999-09-01

    Formic acid has been used in various countries for the control of parasitic mites of honey bees (Apis mellifera), particularly the Varroa mite (Varroa jacobsoni) and the tracheal mite (Acarapis woodi). Its corrosivity and consequent fear of liability have precluded commercial interest in the United States, and its rapid vaporization requires frequent reapplication. We have developed a gel formulation of formic acid which provides controlled release over 2-3 weeks and improves the convenience and safety of handling of formic acid. The strong acidity of formic acid restricts the choice of gelling agents; vegetable gellants such as agar are destroyed, and bentonite clay derivatives do not gel, even with high-shear mixing. Polyacrylamides lead to viscous liquids lacking thixotropic properties. High-molecular-weight poly(acrylic acids) and fumed silicas provided gels with suitable physical characteristics. The poly(acrylic acid) gels were difficult to mix and gave slower and nonlinear release behavior, while the fumed silica gels were easy to prepare and linear in formic acid vaporization.

  19. An inexpensive microslab gel DNA electrophoresis system with real-time fluorescence detection.

    PubMed

    Chen, Xiaojia; Ugaz, Victor M

    2006-02-01

    In this paper, we describe the construction of a simple yet powerful gel electrophoresis apparatus that can be used to perform size-selective separations of DNA fragments in virtually any laboratory. This system employs a microslab gel format with a novel gel casting technique that eliminates the need for delicate combs to define sample loading wells. The compact size of the microslab gel format allows rapid separations to be performed at low voltages using submicroliter sample volumes. Real time fluorescence detection of the migrating DNA fragments is accomplished using an inexpensive digital microscope that directly connects to any PC with a USB interface. The microscope is readily adaptable for this application by replacing its white light source with a blue light-emitting diode (LED) and adding an appropriate emission filter. Both polyacrylamide and agarose gels can be used as separation matrices. Separation performance was characterized using standard dsDNA ladders, and correct sizing of a 191 bp PCR product was achieved in 15 min. The low cost and simplicity of this system makes it ideally suited for use in a variety of laboratory and educational settings.

  20. Synergistic activity between an antimicrobial polyacrylamide and daptomycin versus Staphylococcus aureus biofilm.

    PubMed

    Siala, Wafi; Van Bambeke, Françoise; Taresco, Vincenzo; Piozzi, Antonella; Francolini, Iolanda

    2016-07-01

    Antibiotic resistance of bacteria growing in biofilms compared to their planktonic counterparts enhances the difficulty to eradicate biofilm-associated infections. In the last decade, combination antibiotic therapy has emerged as an attractive strategy for treating biofilm infections, even if in most of tolerant biofilms the optimal combinations are still unknown. In this study, an antimicrobial cationic polyacrylamide was used in combination with daptomycin or moxifloxacin against mature biofilms of Staphylococcus aureus clinical isolates to examine a possible improvement of the antibiofilm activity of the two antibiotics. The polymer did not have an effect on moxifloxacin but significantly increased the antibiofilm efficacy of daptomycin. These findings are presumably related to the different mechanism of action of the two drugs. In summary, our data highlighted the ability of polycations to increase daptomycin antibiofilm activity providing a potential strategy to eradicate biofilms in industrial or medical settings.

  1. Cationic polyacrylamide enhancing cellulase treatment efficiency of hardwood kraft-based dissolving pulp.

    PubMed

    Wang, Qiang; Liu, Shanshan; Yang, Guihua; Chen, Jiachuan; Ni, Yonghao

    2015-05-01

    Cellulase treatment for decreasing viscosity and increasing Fock reactivity of dissolving pulp is a promising approach to reduce the use of toxic chemicals, such as hypochlorite in the dissolving pulp manufacturing process in the industry. Improving the cellulase treatment efficiency during the process is of practical interest. In the present study, the concept of using cationic polyacrylamide (CPAM) to enhance the cellulase treatment efficiency was demonstrated. This was mainly attributed to the increased cellulase adsorption onto cellulose fibers based on the patching/bridging mechanism. Results showed that the cellulase adsorption was increased by about 20% with the addition of 250 ppm of CPAM under the same conditions as those of the control. It was found that the viscosity decrease and Fock reactivity increase for the cellulase treatment was enhanced from using CPAM. The CPAM-assisted cellulase treatment concept may provide a practical alternative to the present hypochlorite-based technology for viscosity control in the industry.

  2. Synthesis and properties of immobilized pectinase onto the macroporous polyacrylamide microspheres.

    PubMed

    Lei, Zhongli; Jiang, Qin

    2011-03-23

    Pectinase was covalently immobilized onto the macroporous polyacrylamide (PAM) microspheres synthesized via an inverse suspension polymerization approach, resulting in 81.7% immobilization yield. The stability of the macroporous PAM support, which has a large surface area, is not impeded by the adsorbed proteins despite the fact that up to 296.3 mg of enzyme is immobilized per gram of the carrier particles. The immobilized enzyme retained more than 75% of its initial activity over 30 days, and the optimum temperature/pH also increased to the range of 50-60 °C/3.0-5.0. The immobilized enzyme also exhibited great operational stability, and more than 75% residual activity was observed after 10 batch reactions. The kinetics of a model reaction catalyzed by the immobilized pectinase was finally investigated. Moreover, the immobilized pectinase could be recovered by centrifuging and showed durable activity at the process of recycle.

  3. Effect of crosslinker length on the elastic and compression modulus of poly(acrylamide) nanocomposite hydrogels

    NASA Astrophysics Data System (ADS)

    Zaragoza, J.; Chang, A.; Asuri, P.

    2017-01-01

    Polymer hydrogelshave shown to exhibit improved properties upon the addition of nanoparticles; however, the mechanical underpinnings behind these enhancements have not been fully elucidated. Moreover, fewer studies have focused on developing an understanding of how polymer parameters affect the nanoparticle-mediated enhancements. In this study, we investigated the elastic properties of silica nanoparticle-reinforced poly(acrylamide) hydrogels synthesized using crosslinkers of various lengths. Crosslinker length positively affected the mechanical properties of hydrogels that were synthesized with or without nanoparticles. However the degree of nanoparticle enhancement was negatively correlated to crosslinker length. Our findings enable the understanding of the respective roles of nanoparticle and polymer properties on nanoparticle-mediated enhancement of hydrogels and thereby the development of next-generation nanocomposite materials.

  4. A fully automated linear polyacrylamide coating and regeneration method for capillary electrophoresis of proteins.

    PubMed

    Bodnar, Judit; Hajba, Laszlo; Guttman, Andras

    2016-12-01

    Surface modification of the inner capillary wall in CE of proteins is frequently required to alter EOF and to prevent protein adsorption. Manual protocols for such coating techniques are cumbersome. In this paper, an automated covalent linear polyacrylamide coating and regeneration process is described to support long-term stability of fused-silica capillaries for protein analysis. The stability of the resulting capillary coatings was evaluated by a large number of separations using a three-protein test mixture in pH 6 and 3 buffer systems. The results were compared to that obtained with the use of bare fused-silica capillaries. If necessary, the fully automated capillary coating process was easily applied to regenerate the capillary to extend its useful life-time.

  5. Direct capture of lactoferrin from cheese whey on supermacroporous column of polyacrylamide cryogel with copper ions.

    PubMed

    Carvalho, B M A; Carvalho, L M; Silva, W F; Minim, L A; Soares, A M; Carvalho, G G P; da Silva, S L

    2014-07-01

    Lactoferrin is a protein that is present in cheese whey (a waste product from the dairy industry) and has several biological activities. However, its production from whey must have a high yield and low cost for industrial applications. As such, this study reports the use of polyacrylamide cryogel, loaded with Cu(2+) (through the bond with iminodiacetic acid (IDA)), as an adsorbent for the chromatographic process to capture lactoferrin whey. Ultrafiltered cheese whey was passed through the cryogel-IDA-Cu(2+) system. The eluates were subjected to analysis of total protein, SDS-PAGE and size exclusion chromatography. The results showed an axial dispersion coefficients, at different superficial velocities of liquid, in a range of 10(-6)-10(-5)m(2)/s. The cryogel demonstrated good hydraulic permeability (4.7086×10(-13)m(2)) and a porosity of approximately 78.2%. The IDA-Cu(2+) cryogel system was also able to capture lactoferrin in high purity.

  6. Polyacrylamide grafting of modified graphene oxides by in situ free radical polymerization

    SciTech Connect

    Tang, Mingyi; Xu, Xiaoyang; Wu, Tao; Zhang, Sai; Li, Xianxian; Li, Yi

    2014-12-15

    Highlights: • Graphene oxide (GO) was modified by chemical reactions to functionalized GO (FGO). • The FGOs and the GO were then subjected to in situ free radical polymerization. • Hydroxyl groups of GO were the most reactive grafting sites. - Abstract: Graphene oxide (GO) was modified using chemical reactions to obtain three types of functionalized GO sheets (FGO). The FGO sheets and the GO were then subjected to in situ free radical polymerization in order to study the grafting polymerization. The FGO and grafted-.FGO were analyzed with Fourier transform infrared spectroscopy, scanning electronic microscopy, thermo-gravimetric analysis (TGA) and X-ray photoelectron spectroscopy (XPS). The grafting percentages in the materials were calculated using the TGA and XPS results. The FGO sheets with different functional groups exhibited different grafting abilities, and hydroxyl groups were proven to be the most reactive grafting sites for the in situ free radical grafting polymerization of polyacrylamide.

  7. Study of polyacrylamide grafted starch based algal flocculation towards applications in algal biomass harvesting.

    PubMed

    Banerjee, Chiranjib; Gupta, Pratibha; Mishra, Sumit; Sen, Gautam; Shukla, Pratyoosh; Bandopadhyay, Rajib

    2012-11-01

    Microalgae may be the source of high amount of lipid and protein. It has the property for carbon dioxide sequestration, recycling and also can remove pollutants from wastewater. Using traditional methods, collection of algal biomass is either cost effective, time consuming or may be toxic due to use of chemical salts. The aim of this study is to harvest freshwater microalgae (Chlorella sp. CB4) biomass by using polymer. Polyacrylamide grafted starch (St-g-PAM) has been synthesized by microwave assisted method involving a synergism of microwave radiation and ceric ammonium nitrate (CAN) to initiate the grafting reaction. The synthesis was optimized in terms of CAN and monomer (acrylamide) concentration. The algal flocculation efficacy of all the grades of this graft copolymer was studied through standard 'Jar test' procedure. Effects of percentage grafting, pH and zeta potential on percentage recovery of algal biomass were thoroughly investigated.

  8. Mercapto functionalized silica entrapped polyacrylamide hydrogel: Arsenic adsorption behaviour from aqueous solution.

    PubMed

    Kumar, Rajesh; Jain, S K; Verma, S; Malodia, P

    2015-10-15

    In this article, 3-mercaptopropyl functionalized silica entrapped polyacrylamide hydrogel (MPFS-PAA) was prepared and characterized by FT-IR, scanning electron microscopy (SEM) and energy dispersion X-ray spectroscopy (EDS). Synthesized hydrogel was evaluated for removal of arsenic(III) from aqueous solution. Adsorption studies were carried out by batch method as function of contact time, initial concentration of arsenic and pH. As(III) adsorption data fitted well with Langmuir and Freundlich isotherm models. Adsorption capacity of arsenic 92.5 μg/g was obtained at initial concentration of 100 μg/L by Langmuir isotherm. Adsorption kinetics was tested for pseudo-second order reaction at different contact time. The rate constants of pseudo second order reaction were calculated and good correlation coefficient R(2) 99.67 obtained. The results indicates that MPFS-PAA is an effective adsorbent for removal of As(III) from aqueous solution.

  9. A rapid and efficient two-step gel electrophoresis method for the purification of major rye grass pollen allergens.

    PubMed

    Levy, D; Davies, J; O'Hehir, R; Suphioglu, C

    2001-06-01

    Purified proteins are mandatory for molecular, immunological and cellular studies. However, purification of proteins from complex mixtures requires specialised chromatography methods (i.e., gel filtration, ion exchange, etc.) using fast protein liquid chromatography (FPLC) or high-performance liquid chromatography (HPLC) systems. Such systems are expensive and certain proteins require two or more different steps for sufficient purity and generally result in low recovery. The aim of this study was to develop a rapid, inexpensive and efficient gel-electrophoresis-based protein purification method using basic and readily available laboratory equipment. We have used crude rye grass pollen extract to purify the major allergens Lol p 1 and Lol p 5 as the model protein candidates. Total proteins were resolved on large primary gel and Coomassie Brilliant Blue (CBB)-stained Lol p 1/5 allergens were excised and purified on a secondary "mini"-gel. Purified proteins were extracted from unstained separating gels and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analyses. Silver-stained SDS-PAGE gels resolved pure proteins (i.e., 875 microg of Lol p 1 recovered from a 8 mg crude starting material) while immunoblot analysis confirmed immunological reactivity of the purified proteins. Such a purification method is rapid, inexpensive, and efficient in generating proteins of sufficient purity for use in monoclonal antibody (mAb) production, protein sequencing and general molecular, immunological, and cellular studies.

  10. Wetting mechanisms of gel-based controlled-release fertilizers.

    PubMed

    Shavit, U; Reiss, M; Shaviv, A

    2003-02-14

    The release mechanism of gel-based controlled release fertilizers (CRFs) involves water penetration into dry mixtures of fertilizers and gel forming polymers. Water penetration provides an upper limit to the whole release process. Where wetting prediction is often based on models that describe the flow of the liquid phase, vapor motion may become significant when a sharp wetting front exists. In this study we examine the role of vapor and fluid flows in the wetting process of CRFs consisting of urea or KNO(3) mixed with polyacrylamide (PAM). Vapor adsorption isotherms were obtained for typical fertilizer-PAM mixtures. Wetting and release experiments were conducted by dividing the CRFs into regions alternately filled with a pure fertilizer and mixtures of PAM and fertilizer. The experiments were designed in such a way that when the wetting front reaches a mixtures interface, its motion depends on the gradient imposed by the difference in osmotic potential (OP). The coupled equations of vapor and liquid flow in initially dry conditions were solved numerically to demonstrate the conceptual understanding gained by the experiments. The results show that wetting front motion is affected by transport and adsorption of vapor. It was also shown that the release rate is different when wetting is governed by vapor flow or by liquid flow. The release pattern from a multi-regions device was consistent with the wetting pattern, demonstrating the possibility to tailor the release according to periods of peak demand.

  11. Silk sericin/polyacrylamide in situ forming hydrogels for dermal reconstruction.

    PubMed

    Kundu, Banani; Kundu, Subhas C

    2012-10-01

    In situ forming tissue sealants are advantageous due to ease in application, complete coverage of defect site and assured comfort levels to patients. The interconnected three-dimensional hydrophilic networks perfectly manage typical dermal wounds by suitably scaffolding skin fibroblast, diffusing the nutrients, therapeutics and exudates while still maintaining an adequately moist environment. We evaluate the cell homing ability of semi-interpenetrating non-mulberry tropical tasar silk sericin/polyacrylamide hydrophilic network with a keen understanding of its network characteristics and correlation of protein concentration with the performance as cell scaffold. Interconnectivity of porous networks observed through scanning electron micrograph revealed pore sizes ranging from 23 to 52 μm. The enhanced β-sheet content with the increasing sericin concentration in far red spectroscopy study supported their corresponding improved compressive strength. These semi-interpenetrating networks were found to possess a maximum fluid uptake of 112% of its weight, hence preventing the accumulation of exudates at the wound area. The present systems appear to possess characteristics like rapid gelation (~5min) at 37 °C, 98% porosity enabling the migration of fibroblasts during healing (observed through confocal and scanning electron micrographs), cell adhesion together with the absence of any cyto-toxic effect suggesting its potential as in situ tissue sealants. The compressive strength up to 61 kPa ensured ease in handling even when wet. The results prove the suitability to use non-mulberry tasar cocoon silk sericin/polyacrylamide semi-interpenetrating network as a reconstructive dermal sealant.

  12. Fate and efficacy of polyacrylamide applied in furrow irrigation: full-advance and continuous treatments.

    PubMed

    Lentz, Rodrick D; Sojka, Robert E; Mackey, Bruce E

    2002-01-01

    Polyacrylamide (PAM) is applied to 400000 irrigated hectares annually in the USA to control irrigation-induced erosion, yet the fate of dissolved PAM applied in irrigation water is not well documented. We determined the fate of PAM added to furrow streams under two treatments: Initial-10, 10 mg L(-1) PAM product applied only during the initial hours of the irrigation, and Cont-1, 1.0 mg L(-1) PAM product applied continuously during the entire irrigation. The study measured PAM concentrations in 167-m-long PAM-treated furrow streams and along a 530-m tail ditch that received this runoff. Soil was Portneuf silt loam (coarse-silty, mixed, superactive, mesic Durinodic Xeric Haplocalcid) with 1.5% slope. Samples were taken at three times during the irrigations, both during and after PAM application. Polyacrylamide was adsorbed to soil and removed from solution as the streams traversed the soil-lined channels. The removal rate increased with stream sediment concentration. Stream sediment concentrations were higher when PAM concentrations were <2 mg L(-1) a.i., for early irrigations, and when untreated tributary flows combined with the stream. In these cases, PAM concentration decreased to undetectable levels over the flow lengths used in this study. When inflows contained >6 mg L(-1) PAM a.i., stream sediment concentrations were minimal and PAM concentrations did not change down the furrow, though they decreased to undetectable levels within 0.5 h after application ceased. One percent of applied PAM was lost in tail-ditch runoff. This loss could have been eliminated by treating only the furrow advance or not treating the last two irrigations.

  13. An NMR relaxometry and gravimetric study of gelatin-free aqueous polyacrylamide dosimeters

    NASA Astrophysics Data System (ADS)

    Babic, Steven; Schreiner, L. John

    2006-09-01

    In conformal radiation therapy, a high dose of radiation is given to a target volume to increase the probability of cure, and care is taken to minimize the dose to surrounding healthy tissue. The techniques used to achieve this are very complicated and the precise verification of the resulting three-dimensional (3D) dose distribution is required. Polyacrylamide gelatin (PAG) dosimeters with magnetic resonance imaging and optical computed tomography scanning provide the required 3D dosimetry with high spatial resolution. Many basic studies have characterized these chemical dosimeters that polymerize under irradiation. However, the investigation of the fundamental properties of the radiation-induced polymerization in PAG dosimeters is complicated by the presence of the background gelatin matrix. In this work, a gelatin-free model system for the study of the basic radiation-induced polymerization in PAG dosimeters has been developed. Experiments were performed on gelatin-free dosimeters, named aqueous polyacrylamide (APA) dosimeters, containing equal amounts of acrylamide and N,N'-methylene-bisacrylamide. The APA dosimeters were prepared with four different total monomer concentrations (2, 4, 6 and 8% by weight). Nuclear magnetic resonance (NMR) spin-spin and spin-lattice proton relaxation measurements at 20 MHz, and gravimetric analyses performed on all four dosimeters, show a continuous degree of polymerization over the dose range of 0-25 Gy. The developed NMR model explains the relationship observed between the relaxation data and the amount of crosslinked polymer formed at each dose. This model can be extended with gelatin relaxation data to provide a fundamental understanding of radiation-induced polymerization in the conventional PAG dosimeters.

  14. Assessment of segmental myocardial viability using regional 2-dimensional strain echocardiography.

    PubMed

    Migrino, Raymond Q; Zhu, Xiaoguang; Pajewski, Nicholas; Brahmbhatt, Tejas; Hoffmann, Raymond; Zhao, Ming

    2007-04-01

    We determined whether 2-dimensional strain echocardiography can identify viable from infarcted myocardium in a rat ischemia-reperfusion model. A total of 16 male Sprague-Dawley rats underwent left anterior descending coronary artery occlusion for 12 or 30 minutes followed by 60-minute reperfusion. Short-axis 2-dimensional strain echocardiography was performed at the mid-ventricle 60 minutes post-reperfusion. Post-sacrifice, triphenyl tetrazolium chloride was infused to the coronary circulation. Regional end-systolic radial and circumferential strain, and time to peak strain, were measured using software in all 96 segments and correlated with areas of infarct in corresponding histologic slices. Segments with greater than 50% area of infarct had lower end-systolic radial and circumferential strain and longer time to peak strain versus areas with 50% or less strain or no infarct. Extent of infarct correlates with radial and circumferential strain. End-systolic radial strain less than 2% has 88% sensitivity and 95% specificity for detecting infarcted area greater than 50%. Two-dimensional strain echocardiography-derived strain is useful in distinguishing infarcted from viable myocardium.

  15. Determining the Best Sensing Coverage for 2-Dimensional Acoustic Target Tracking

    PubMed Central

    Pashazadeh, Saeid; Sharifi, Mohsen

    2009-01-01

    Distributed acoustic target tracking is an important application area of wireless sensor networks. In this paper we use algebraic geometry to formally model 2-dimensional acoustic target tracking and then prove its best degree of required sensing coverage. We present the necessary conditions for three sensing coverage to accurately compute the spatio-temporal information of a target object. Simulations show that 3-coverage accurately locates a target object only in 53% of cases. Using 4-coverage, we present two different methods that yield correct answers in almost all cases and have time and memory usage complexity of Θ(1). Analytic 4-coverage tracking is our first proposed method that solves a simultaneous equation system using the sensing information of four sensor nodes. Redundant answer fusion is our second proposed method that solves at least two sets of simultaneous equations of target tracking using the sensing information of two different sets of three sensor nodes, and fusing the results using a new customized formal majority voter. We prove that 4-coverage guarantees accurate 2-dimensional acoustic target tracking under ideal conditions. PMID:22412319

  16. Crossover from 2-dimensional to 3-dimensional aggregations of clusters on square lattice substrates

    NASA Astrophysics Data System (ADS)

    Cheng, Yi; Zhu, Yu-Hong; Pan, Qi-Fa; Yang, Bo; Tao, Xiang-Ming; Ye, Gao-Xiang

    2015-11-01

    A Monte Carlo study on the crossover from 2-dimensional to 3-dimensional aggregations of clusters is presented. Based on the traditional cluster-cluster aggregation (CCA) simulation, a modified growth model is proposed. The clusters (including single particles and their aggregates) diffuse with diffusion step length l (1 ≤ l ≤ 7) and aggregate on a square lattice substrate. If the number of particles contained in a cluster is larger than a critical size sc, the particles at the edge of the cluster have a possibility to jump onto the upper layer, which results in the crossover from 2-dimensional to 3-dimensional aggregations. Our simulation results are in good agreement with the experimental findings. Project supported by the National Natural Science Foundation of China (Grant Nos. 11374082 and 11074215), the Science Foundation of Zhejiang Province Department of Education, China (Grant No. Y201018280), the Fundamental Research Funds for Central Universities, China (Grant No. 2012QNA3010), and the Specialized Research Fund for the Doctoral Program of Higher Education of China (Grant No. 20100101110005).

  17. Long-term polyacrylamide formulation effects on soil erosion, water infiltration, and yields of furrow-irrigated crops

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Water soluble anionic polyacrylamide (WSPAM) is an important tool employed by producers to reduce erosion associated with furrow irrigation. Two formulations of WSPAM are used in agriculture, although few if any reports comparing their effectiveness are published. A long-term field study was condu...

  18. Removal of paraquat pesticide from aqueous solutions using a novel adsorbent material based on polyacrylamide and methylcellulose hydrogels

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This research studied the characteristics of poly(acrylamide) and methylcellulose (PAAm-MC) hydrogels as a novel adsorbent material for removal of pesticide paraquat, from aqueous solution, with potential applications in curbing environmental risk from such herbicides. PAAm-MC hydrogels with differe...

  19. Development of Polymer Gel Systems to Improve Volumetric Sweep and Reduce Producing Water/Oil Ratios

    SciTech Connect

    G. Paul Willhite; Stan McCool; Don W. Green; Min Cheng; Feiyan Chen

    2005-12-31

    Gelled polymer treatments are applied to oil reservoirs to increase oil production and to reduce water production by altering the fluid movement within the reservoir. This report describes the results of a 42-month research program that focused on the understanding of gelation chemistry and the fundamental mechanisms that alter the flows of oil and water in reservoir rocks after a gel treatment. Work was conducted on a widely applied system in the field, the partially hydrolyzed polyacrylamide-chromium acetate gel. Gelation occurs by network formation through the crosslinking of polyacrylamide molecules as a result of reaction with chromium acetate. Pre-gel aggregates form and grow as reactions between chromium acetate and polyacrylamide proceed. A rate equation that describes the reaction between chromium acetate and polymer molecules was regressed from experimental data. A mathematical model that describes the crosslinking reaction between two polymer molecules as a function of time was derived. The model was based on probability concepts and provides molecular-weight averages and molecular-weight distributions of the pre-gel aggregates as a function of time and initial system conditions. Average molecular weights of pre-gel aggregates were measured as a function of time and were comparable to model simulations. Experimental methods to determine molecular weight distributions of pre-gel aggregates were unsuccessful. Dissolution of carbonate minerals during the injection of gelants causes the pH of the gelant to increase. Chromium precipitates from solution at the higher pH values robbing the gelant of crosslinker. Experimental data on the transport of chromium acetate solutions through dolomite cores were obtained. A mathematical model that describes the transport of brine and chromium acetate solutions through rocks containing carbonate minerals was used to simulate the experimental results and data from literature. Gel treatments usually reduce the permeability

  20. Polyoxometalate-based Supramolecular Gel

    NASA Astrophysics Data System (ADS)

    He, Peilei; Xu, Biao; Liu, Huiling; He, Su; Saleem, Faisal; Wang, Xun

    2013-05-01

    Self-assemblyings of surfactant-encapsulated Wells-Dawson polyoxometalates (SEPs) nanobuilding blocks in butanone and esters yielded supramolecular gels showing thermo and photo responsive properties. The gels can be further polymerized if unsaturated esters were used and subsequently electrospinned into nanowires and non-woven mats. The as-prepared non-woven mats have a Young's modulus as high as 542.55 MPa. It is believed that this supramolecular gel is a good platform for polyoxometalates processing.

  1. Prenatal 2-dimensional and 3-dimensional ultrasonography diagnosis and autoptic findings of isolated ectopia cordis.

    PubMed

    Bianca, S; Bartoloni, G; Auditore, S; Reale, A; Tetto, C; Ingegnosi, C; Pirruccello, B; Ettore, G

    2006-01-01

    Ectopia cordis is a very rare congenital malformation, commonly associated with intracardiac anomalies. It is due to a defect in fusion of the anterior chest wall resulting in an extrathoracic location of the heart. We report prenatal 2-dimensional (2D) and 3D ultrasonography diagnosis and postnatal autoptic findings of an isolated ectopia cordis with tricuspid atresia. Ectopia cordis prenatal diagnosis is easily made with ultrasound by visualizing the heart outside the thoracic cavity. 3D ultrasonography may add more detailed visualization of the heart anomaly even if the 2D ultrasonography alone permits the prenatal diagnosis. Obstetrical management should include a careful search for associated anomalies, especially cardiac, and the assessment of fetal karyotype. As this is considered a sporadic anomaly, the recurrence risk is low and no genetic origin is known.

  2. Determination of chemical concentration with a 2 dimensional CCD array in the Echelle grating spectrometer

    SciTech Connect

    Lewis, D.K.; Stevens, C.G.

    1994-11-15

    The Echelle grating spectrometer (EGS) uses a stepped Echelle grating, prisms and a folded light path to miniaturize an infrared spectrometer. Light enters the system through a slit and is spread out along Y by a prism. This light then strikes the grating and is diffracted out along X. This spreading results in a superposition of spectral orders since the grating has a high spectral range. These orders are then separated by again passing through a prism. The end result of a measurement is a 2 dimensional image which contains the folded spectrum of the region under investigation. The data lies in bands from top to bottom, for example, with wavenumber increments as small as 0.1 lying from left to right such that the right end of band N is the same as the left end of band N+1. This is the image which must be analyzed.

  3. Gel polymer electrolytes for batteries

    DOEpatents

    Balsara, Nitash Pervez; Eitouni, Hany Basam; Gur, Ilan; Singh, Mohit; Hudson, William

    2014-11-18

    Nanostructured gel polymer electrolytes that have both high ionic conductivity and high mechanical strength are disclosed. The electrolytes have at least two domains--one domain contains an ionically-conductive gel polymer and the other domain contains a rigid polymer that provides structure for the electrolyte. The domains are formed by block copolymers. The first block provides a polymer matrix that may or may not be conductive on by itself, but that can soak up a liquid electrolyte, thereby making a gel. An exemplary nanostructured gel polymer electrolyte has an ionic conductivity of at least 1.times.10.sup.-4 S cm.sup.-1 at 25.degree. C.

  4. Highly sensitive immunoassay based on controlled rehydration of patterned reagents in a 2-dimensional paper network.

    PubMed

    Fridley, Gina E; Le, Huy; Yager, Paul

    2014-07-01

    We have demonstrated a multistep 2-dimensional paper network immunoassay based on controlled rehydration of patterned, dried reagents. Previous work has shown that signal enhancement improves the limit of detection in 2-dimensional paper network assays, but until now, reagents have only been included as wet or dried in separate conjugate pads placed at the upstream end of the assay device. Wet reagents are not ideal for point-of-care because they must be refrigerated and typically limit automation and require more user steps. Conjugate pads allow drying but do not offer any control of the reagent distribution upon rehydration and can be a source of error when pads do not contact the assay membrane uniformly. Furthermore, each reagent is dried on a separate pad, increasing the fabrication complexity when implementing multistep assays that require several different reagents. Conversely, our novel method allows for consistent, controlled rehydration from patterned reagent storage depots directly within the paper membrane. In this assay demonstration, four separate reagents were patterned in different regions of the assay device: a gold-antibody conjugate used for antigen detection and three different signal enhancement components that must not be mixed until immediately before use. To show the viability of patterning and drying reagents directly onto a paper device for dry reagent storage and subsequent controlled release, we tested this device with the malaria antigen Plasmodium falciparum histidine-rich protein 2 (PfHRP2) as an example of target analyte. In this demonstration, the signal enhancement step increases the visible signal by roughly 3-fold and decreases the analytical limit of detection by 2.75-fold.

  5. Fetal Cerebellar Vermis Circumference Measured by 2-Dimensional Ultrasound Scan: Reference Range, Feasibility and Reproducibility

    PubMed Central

    Spinelli, M.; Sica, C.; Meglio, L. D.; Bolla, D.; Raio, L.; Surbek, D.

    2016-01-01

    Purpose: To provide 2-dimensional ultrasonographic (2D-US) normograms of cerebellar vermis biometry, as well as to evaluate the feasibility and the reproducibility of these measurements in clinical practice. Materials and Methods: A prospective cross-sectional study of 328 normal singleton pregnancies between 18 and 33 weeks of gestation. Measurements of the fetal cerebellar vermis circumference (VC) in the mid-sagittal plane were performed by both a senior and a junior operator using 2D-US. VC as a function of gestational age (GA) was expressed by regression equations. In 24 fetuses 3-dimensional (3D) reconstructed planes were obtained in order to allow comparisons with 2D-US measurements. The agreement between 2D and 3D measurements and the interobserver variability were assessed by interclass correlation coefficients (ICC). Results: Satisfactory vermis measurements could be obtained in 89.9% of cases. The VC (constant= − 12.21; slope=2.447; r=0.887, p<0.0001) correlated linearly with GA. A high degree of consistency was observed between 2D and 3D ultrasound measurements (ICC=0.846 95% CI 679–0.930) as well as between measurements obtained by different examiners (ICC=0.890 95% CI 989–0.945). Conclusion: 2-dimensional ultrasonographic measurements of cerebellar vermis throughout gestation in the mid-sagittal view seem to be feasible and reproducible enough to be potentially used in clinical practice. Such measurements may supply a tool for accurate identification of posterior fossa anomalies, providing the basis for proper counseling and management and of these conditions. PMID:27921094

  6. Polyacrylamide effect on hydraulic conductivity of hardsetting soils in Northeast of Brazil

    NASA Astrophysics Data System (ADS)

    Silva, Laércio; Almeida, Brivaldo; Melo, Diego; Marques, Karina; Almeida, Ceres

    2013-04-01

    Among soil hydro-physical properties, hydraulic conductivity is more sensitive to changes in soil structure. Hydraulic conductivity describes the ease with which a fluid (usually water) can move through pore spaces or fractures. It depends on the intrinsic permeability of the material and on the degree of saturation, and on the density and viscosity of the fluid. Hardsetting soils present very low hydraulic conductivity values. When dry, these soils show high penetration resistance and consistency extremely hard, but change to friable when moist. In this condition are poorly structured, slaking when moist, limit agricultural machinery use and it may reduce the growth of the root system. In Brazil, these soils occur throughout of coastal zone in flat areas called "coastal tableland". Chemical ameliorant, such as polymers based on anionic polyacrylamide (PAM), improve hydraulic conductivity of soil in hardsetting soils. The primary functions of polyacrylamide soil conditioners are to increase soil tilth, aeration, and porosity and reduce compaction and water run-off. PAM effect is attributed to its ability to expand when placed in water, storing it in soil pore space, releasing it gradually to the plants. This process occurs by reducing the water flow through the pores of the soil, due to water molecules can be absorbed by PAM, providing water gradually. Thus, this study tested the hypothesis that PAM reduces the soil hardsetting character. The area is located in coastal zone in Goiana city, Pernambuco, northeastern of Brazil. This soil is typical hardsetting soil. Intact soil cores were collected from four horizons until 70cm depth. In the laboratory, the soil cores were saturated with different PAM concentrations (0.01, 0.005, 0.00125%) and H2O (control). Saturated hydraulic conductivity (Ksat) was determined using a constant head method, according to Klute and Dirksen (1986). Four replicates were used for each horizon and Tukey test at 5% probability was used by

  7. Impact of reaction conditions on architecture and rheological properties of starch graft polyacrylamide polymers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We carried out experiments examining the impact that solvent selection and reaction conditions have on the radical initiated graft polymerization reaction of acrylamide onto starch. We have also evaluated the rheological properties the starch graftpolyacrylamide product when a gel is formed in water...

  8. Mobility shift detection of phosphorylation on large proteins using a Phos-tag SDS-PAGE gel strengthened with agarose.

    PubMed

    Kinoshita, Eiji; Kinoshita-Kikuta, Emiko; Ujihara, Hiromi; Koike, Tohru

    2009-08-01

    We describe a novel technique of phosphate-affinity SDS-PAGE using Phos-tag to analyze large phosphoproteins with molecular masses of more than 200 kDa. The protein phosphoisotypes were clearly separated as up-shifted migration bands in a 3% w/v polyacrylamide gel containing 20 microM Phos-tag and 0.5% w/v agarose. In subsequent immunoblotting, the procedure permitted the determination of the phosphoisotypes of high-molecular-mass proteins, such as mTOR (289 kDa), ATM kinase (350 kDa), and 53BP1 (213 kDa).

  9. Characteristics of a novel polymer gel dosimeter formula for MRI scanning: Dosimetry, toxicity and temporal stability of response.

    PubMed

    Abtahi, S M

    2016-09-01

    The present study intended to investigate the composition of a new polymer gel dosimeter. The new composition would be more suitable for a wide range of applications in comparison to polyacrylamide gel dosimeter since its extremely toxic acrylamide has been replaced with less harmful monomer i.e. 2-Acrylamido-2-MethylPropane Sulfonic acid (AMPS). To this end, the PAGAT gel dosimeter formula was used as a basis to test the new formulation of polymer gel dosimeter with a different monomer (AMPS) instead of acrylamide by using the %6T and %50C to the formula. The new formulation was named PAMPSGAT (Poly AMPS, Gelatin and THPC) polymer gel dosimeter. Moreover, the MRI response (R2) of dosimeters was analyzed in terms of different dose range as well as post-irradiation time. The results indicated that the dose-response (R2) of AMPS/Bis had a linear trend over a wide dose range. Furthermore, the results showed an acceptable temporal stability for the new polymer gel dosimeter.

  10. A new injectable biphasic hydrogel based on partially hydrolyzed polyacrylamide and nano hydroxyapatite, crosslinked with chromium acetate, as scaffold for cartilage regeneration

    NASA Astrophysics Data System (ADS)

    Koushki, N.; Tavassoli, H.; Katbab, A. A.; Katbab, P.; Bonakdar, S.

    2015-05-01

    Polymer scaffolds are applied in the field of tissue engineering as three dimensional structures to organize cells and present stimuli to direct generation of a desired damaged tissue. In situ gelling scaffolds have attracted great attentions, as they are structurally similar to the extra cellular matrix (ECM). In the present work, attempts have been made to design and fabricate a new injectable and crosslinkable biphasic hydrogel based on partially hydrolyzed polyacrylamide (HPAM), chromium acetate as crosslink agent and nanocrystalline hydroxyapatite (nHAp) as reinforcing and bioactive agent for repair and regeneration of damaged cartilage. The distinct characteristic of HPAM is the presence of carboxylate anion groups on its backbone which allows to engineer the structure of the hydrogel for the desired bioactivity with appropriate cells differentiation towards both soft and hard (bone) tissues. The synthesized hydrogel exhibited bifunctional behavior which was derived by its biphasic structure in which one phase was loaded with nano hydroxyapatite to provide integration capability by subchondral bones and fix the hydrogel at cartilage defect without a need for suturing. The other phase differentiates the rabbit adipogenic mesenchymal stem cells (MSCs) towards soft tissue. Rheomechanical spectrometry (RMS) was employed to study the kinetic of the gelation including induction time and rate, as well as to measure the ultimate elastic modulus of the optimum crosslinked hydrogel. Surface tension measurement was also performed to tailor the surface characteristics of the gels. In vitro culturing of the cells inside the crosslinked hydrogel revealed high viability and high differentiation of the encapsulated rabbit stem cells, providing that the chromium acetate level was kept below 0.2 wt%. Based on the obtained results, the designed and fabricated biphasic hydrogel exhibited high potential as carrier for the stem cells for cartilage tissue engineering application

  11. Homogeneity of gels and gel-derived glasses

    NASA Technical Reports Server (NTRS)

    Mukherjee, S. P.

    1984-01-01

    The significance and implications of gel preparation procedures in controlling the homogeneity of multicomponent oxide gels are discussed. The role of physicochemical factors such as the structure and chemical reactivities of alkoxides, the formation of double-metal alkoxides, and the nature of solvent(s) are critically analyzed in the context of homogeneity of gels during gelation. Three procedures for preparing gels in the SiO2-B2O3-Na2O system are examined in the context of cation distribution. Light scattering results for glasses in the SiO2-B2O3-Na2O system prepared by both the gel technique and the conventional technique are examined.

  12. Difference gel electrophoresis (DIGE) using CyDye DIGE fluor minimal dyes.

    PubMed

    Chakravarti, Bulbul; Gallagher, Sean R; Chakravarti, Deb N

    2005-02-01

    One- and two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (1- and 2-D SDS-PAGE) have been widely used for the separation and quantitative estimation of proteins. Following electrophoresis, the gels are stained appropriately to visualize the proteins. Difference gel electrophoresis (DIGE) is a new technique in which different protein samples, individually labeled with specific CyDyes, are combined together followed by electrophoresis and post electrophoretic co-detection and co-analysis on the same gel. CyDye DIGE fluor minimal dyes, which consist of three different CyDyes with different spectral characteristics, have been widely used for such purposes. The technique is highly sensitive with a wide dynamic range for detection of proteins and compatible with state-of-the-art protein identification techniques using mass spectrometry. Although DIGE is mainly used to compare differential expression of various protein samples using 2-D SDS-PAGE, 1-D DIGE also has important applications in quantitative proteomic studies.

  13. Heterogeneity of mammalian DNA ligase detected on activity and DNA sequencing gels.

    PubMed Central

    Mezzina, M; Sarasin, A; Politi, N; Bertazzoni, U

    1984-01-01

    A new method to detect DNA ligase activity in situ after NaDodSO4 polyacrylamide gel electrophoresis has been developed. After renaturation of active polypeptides the ligase reaction occurs in situ by incubating the intact gel in the presence of Mg++ and ATP. Further treatment with alkaline phosphatase removes the unligated 5'-32P-end of oligo (dT) used as a substrate and active polypeptides having ligase activity are identified by autoradiography. Analysis on DNA sequencing gels of the oligo (dT) reaction products present in the activity bands ensures that the radioactive material detected in activity gels or in standard in vitro ligase assays corresponds unambiguously to a ligase activity. Using these methods, we have analysed the purified phage T4 DNA ligase, and the activities present in crude extracts and in purified fractions from monkey kidney (CV1-P) cells. The purified T4 enzyme yields one or two active peptides with Mr values of 60,000 and 70,000. Crude extracts from CV1-P cells contain several polypeptides having DNA ligase activity. Partial purification of these extracts shows that DNA ligase I isolated from hydroxylapatite column is enriched in polypeptides with Mr 200,000, 150,000 and 120,000, while DNA ligase II is enriched in those with Mr 60,000 and 70,000. Images PMID:6377238

  14. A simple monolithic column electroelution for protein recovery from gel electrophoresis.

    PubMed

    Li, Guo-Qing; Shao, Jing; Guo, Chen-Gang; Dong, Jing-Yu; Fan, Liu-Yin; Cao, Cheng-Xi

    2012-11-01

    Protein recovery from gel electrophoresis plays an important role in functional genomics and proteomics but faces a series of issues (e.g., complex procedure, low recovery, long experimental time). In this study, a monolithic column electroelution (MCE) was developed for protein recovery from gel electrophoresis. With the model proteins of bovine serum albumin (BSA), hemoglobin (Hb), and myoglobin (Mb), the developed device and method were compared with common electroelution procedures in agarose gel electrophoresis (AGE). The comparative experiments revealed that (i) the protein recovery achieved with the developed device was greater than 83%, much higher than the 41% to 50% achieved with the common devices; (ii) the running time to obtain 70% recovery was approximately 15 min, evidently shorter than the 240 min with the common devices; and (iii) the device and procedure were simple and less time-consuming as compared with those of the common devices. It was observed that the serum protein bands cut from polyacrylamide gel electrophoresis could be transferred into solution in 15 to 30 min with 82% yield. The device, along with its relevant procedure, has potential use in protein extraction and proteomics as well as in DNA studies.

  15. Agarose gel shift assay reveals that calreticulin favors substrates with a quaternary structure in solution.

    PubMed

    Boelt, Sanne Grundvad; Houen, Gunnar; Højrup, Peter

    2015-07-15

    Here we present an agarose gel shift assay that, in contrast to other electrophoresis approaches, is loaded in the center of the gel. This allows proteins to migrate in either direction according to their isoelectric points. Therefore, the presented assay enables a direct visualization, separation, and prefractionation of protein interactions in solution independent of isoelectric point. We demonstrate that this assay is compatible with immunochemical methods and mass spectrometry. The assay was used to investigate interactions with several potential substrates for calreticulin, a chaperone that is involved in different biological aspects through interaction with other proteins. The current analytical assays used to investigate these interactions are mainly spectroscopic aggregation assays or solid phase assays that do not provide a direct visualization of the stable protein complex but rather provide an indirect measure of interactions. Therefore, no interaction studies between calreticulin and substrates in solution have been investigated previously. The results presented here indicate that calreticulin has a preference for substrates with a quaternary structure and primarily β-sheets in their secondary structure. It is also demonstrated that the agarose gel shift assay is useful in the study of other protein interactions and can be used as an alternative method to native polyacrylamide gel electrophoresis.

  16. Effect of multivalent salts -- calcium and aluminum -- on the flocculation of kaolin suspension with anionic polyacrylamide

    SciTech Connect

    Peng, F.F.; Di, P. . Dept. of Mineral Processing Engineering)

    1994-04-01

    The effects of calcium and aluminum ions on the interaction behavior of anionic polyacrylamide (anionic PAM or HPAM) and kaolin are important in determining the efficient flocculation of kaolin. While kaolin surfaces are negatively charged in media such as water, they exhibit the positive surface charge characteristic at pH below 3.2 in the solution of calcium cation, and at pH below 9.2 in the solution of aluminum cation. The. experimental results show that both calcium and aluminum ions suppress the kaolin flocculation process. The results are analyzed and explained by molecular orbital theory, solution chemistry, infrared spectra, and electronic probe examination of kaolin treated with and without anionic PAM. Analysis results indicate that the specific adsorption of Ca[sup 2+], Al[sup 3+] and their hydroxo complexes on anionic PAM causes the loss of anionic character in the low pH range and leads to a low flocculation efficiency. In the high pH range, the poor kaolin flocculation can be attributed to precipitation of calcium and aluminum hydroxides on active functional groups, which inhibits the hydrogen bonding between anionic PAM and kaolin surfaces. At neutral pH values, the trivalent aluminum ion has more significant adverse effect on the kaolin flocculation than the divalent calcium ion.

  17. Polyacrylamide-clinoptilolite/Y-zeolite composites: characterization and adsorptive features for terbium.

    PubMed

    Baybaş, Demet; Ulusoy, Ulvi

    2011-03-15

    The composites of natural (clinoptilolite) and synthetic zeolite (Z and YZ) with polyacrylamide (PAAm) were synthesized and characterized by FT-IR, TGA, XRD, SEM and PZC analysis. The adsorptive features of the minerals and its composites were investigated for Tb analogues to the rare earth elements (REs) by isotopic tracer method, (160)Tb was the radiotracer. The composites were the hybrid formations of PAAm and Z or YZ. Tb adsorption capacities of the composites were higher than those of bare Z and YZ. The compatibility of Tb adsorption kinetics to the second order and Weber-Morris models implied that the sorption process was chemical via ion exchange. The values of enthalpy and entropy changes were positive and the negative free enthalpy change was evidence for the spontaneity of adsorption. The reusability tests for the composites for five uses demonstrated that the adsorbents could be reused after complete recovery of the loaded ion. Unlike PAAm-YZ, PAAm-Z was resistant to acidic environment. The overall results eventually suggested that the composite of Z and PAAm was a potential cost effective adsorbent for Tb(3+) and REs.

  18. Biodegradation of polyacrylamide by bacteria isolated from activated sludge and oil-contaminated soil.

    PubMed

    Wen, Qinxue; Chen, Zhiqiang; Zhao, Ye; Zhang, Huichao; Feng, Yujie

    2010-03-15

    Polyacrylamide (PAM), a linear water soluble polymeric compound with high molecular weight, is extensively used for oil production in China. Compared with the physico-chemical degradation of PAM, there is no acrylamide monomer, which causes peripheral neuropathy, released in the process of biodegradation. Unfortunately, few microorganisms have been isolated which can degrade PAM. In this study, two PAM-degrading bacterial strains, named HWBI and HWBII, were isolated from the activated sludge and soil in an oil field that had been contaminated by PAM for an extended period. These were subsequently identified as Bacillus cereus and Bacillus flexu, respectively. Both strains grew on a medium composed of 60 mg L(-1) PAM as the sole source of carbon. Although both strains degraded PAM in different rates, after 72 h cultivation more than 70% of the PAM was consumed. This degradation efficiency was much higher than previous studies. Both strains degraded a determinate proportion of PAM when 50-1000 mg L(-1) of the initial PAM was supplied. Glucose with a concentration lower than 200 mg L(-1) can be used as co-metabolism substrate with PAM. The Fourier Transform Infrared (FT-IR) spectrograms of the cultures before and after PAM degradation were also recorded. The result showed that amido groups of the PAM were picked off by the microorganisms from the main chain of the PAM, and metabolism products other than acrylamide were formed in the degradation.

  19. Self-healable, tough, and ultrastretchable nanocomposite hydrogels based on reversible polyacrylamide/montmorillonite adsorption.

    PubMed

    Gao, Guorong; Du, Gaolai; Sun, Yuanna; Fu, Jun

    2015-03-04

    Nanocomposite hydrogels with unprecedented stretchability, toughness, and self-healing have been developed by in situ polymerization of acrylamide with the presence of exfoliated montmorillonite (MMT) layers as noncovalent cross-linkers. The exfoliated MMT clay nanoplatelets with high aspect ratios, as confirmed by transmission electron microscopy (TEM) and X-ray diffraction (XRD) results, are well dispersed in the polyacrylamide matrix. Strong polymer/MMT interaction was confirmed by Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). The effective cross-link densities of these hydrogels are estimated in the range of 2.2-5.7 mol m(-3). Uniaxial tensile tests showed a very high fracture elongation up to 11 800% and a fracture toughness up to 10.1 MJ m(-3). Cyclic loading-unloading tests showed remarkable hysteresis, which indicates energy dissipation upon deformation. Residual strain after cyclic loadings could be recovered under mild conditions, with the recovery extent depending on clay content. A mechanism based on reversible desorption/adsorption of polymer chains on clay platelets surface is discussed. Finally, these nanocomposite hydrogels are demonstrated to fully heal by dry-reswell treatments.

  20. QbD based synthesis and characterization of polyacrylamide grafted corn fibre gum.

    PubMed

    Singh, Akashdeep; Mangla, Bhumika; Sethi, Sheshank; Kamboj, Sunil; Sharma, Radhika; Rana, Vikas

    2017-01-20

    The aim of present investigation was to utilize quality by design approach for the synthesis of polyacrylamide corn fibre gum (PAAm-g-CFG) from corn fibre gum (CFG) by varying concentration of acrylamide and initiator. The spectral analysis (ATR-FTIR, (1)H NMR, DSC, X-ray and Mass spectroscopy) was conducted to assure grafting copolymerization of CFG with acrylamide. The powder flow properties confirm the porous nature of PAAm-g-CFG. The grafted copolymer dispersion showed shear thinning behaviour that follows Herschel Bulkley model. The viscoelastic analysis suggested viscous liquid like nature of PAAm-g-CFG and its viscosity increases with increase in concentration of PAAm-g-CFG. The mucoadhesive strength of synthesized PAAm-g-CFG was found to be higher than moringa oleifera gum, karaya gum, guar gum, xanthan gum, chitosan and gelatin. Further, the results pointed toward enhanced thermal stability of PAAm-g-CFG. Thus, PAAm-g-CFG has a great potential to be used in food and pharmaceutical industry.

  1. A new inorganic-organic composite coagulant, consisting of polyferric sulphate (PFS) and polyacrylamide (PAA).

    PubMed

    Moussas, P A; Zouboulis, A I

    2009-08-01

    Currently, research is focused on the synthesis of new composite coagulants, which are constituted of both inorganic and organic materials. In this paper, the development of relevant reagents was investigated, by combining the inorganic pre-polymerised iron-based coagulant Polyferric Sulphate (PFS) with an organic, non-ionic polymer (Polyacrylamide, PAA) under different PAA/Fe (mg/l) and OH/Fe molar ratios. Moreover, the new reagents were characterised in terms of typical properties, stability and morphological analysis (XRD, FTIR, SEM). Their coagulation performance, when treating low or high turbid kaolin-humic acid suspensions, was also investigated, whereas the applied coagulation mechanisms were discussed by using the Photometric Dispersion Analysis (PDA) analysis. The results show that the new coagulation reagents present improved properties, including increased effective polymer species concentration, and they exhibit very good stability. The respective tests using PDA confirmed that the predominant coagulation mechanism of PFS-PAA is the bridge formation mechanism. Coagulation experiments in low or high turbid kaolin-humic acid suspensions reveal that the novel composite reagent PFS-PAA exhibits better coagulation performance, when compared with simple PFS, in terms of zeta-potential reduction, turbidity and organic matter removal and residual iron concentration.

  2. Hydroxypropyl methyl cellulose grafted with polyacrylamide: application in controlled release of 5-amino salicylic acid.

    PubMed

    Das, Raghunath; Pal, Sagar

    2013-10-01

    In the present study, hydroxypropyl methyl cellulose grafted with polyacrylamide (HPMC-g-PAM) hydrogel was evaluated in vitro as a potential carrier for controlled release of 5-amino salicylic acid (5-ASA). The graft copolymer was developed by grafting PAM chains onto HPMC backbone using potassium persulphate as initiator. The swelling behaviour of hydrogel based tablet was investigated as a function of pH and time in various buffer solutions similar to that of gastric and intestinal fluids. The % equilibrium swelling was found to be higher in case of simulated intestinal fluid (pH=7.4) and lower in simulated gastric fluid (pH=1.2), making an ideal matrix as required for colon specific drug delivery. The drug release study was performed at various pH values akin to the condition of GI tract. The release kinetics of 5-ASA showed non-Fickian diffusion behaviour. This indicates that the release is controlled by a combination of polymer relaxation or erosion of the matrix and diffusion of the drug from the swollen matrix.

  3. Polyacrylamide application versus forest residue mulching for reducing post-fire runoff and soil erosion.

    PubMed

    Prats, Sergio Alegre; Martins, Martinho António Dos Santos; Malvar, Maruxa Cortizo; Ben-Hur, Meni; Keizer, Jan Jacob

    2014-01-15

    For several years now, forest fires have been known to increase overland flow and soil erosion. However, mitigation of these effects has been little studied, especially outside the USA. This study aimed to quantify the effectiveness of two so-called emergency treatments to reduce post-fire runoff and soil losses at the microplot scale in a eucalyptus plantation in north-central Portugal. The treatments involved the application of chopped eucalyptus bark mulch at a rate of 10-12 Mg ha(-1), and surface application of a dry, granular, anionic polyacrylamide (PAM) at a rate of 50 kg ha(-1). During the first year after a wildfire in 2010, 1419 mm of rainfall produced, on average, 785 mm of overland flow in the untreated plots and 8.4 Mg ha(-1) of soil losses. Mulching reduced these two figures significantly, by an average 52 and 93%, respectively. In contrast, the PAM-treated plots did not differ from the control plots, despite slightly lower runoff but higher soil erosion figures. When compared to the control plots, mean key factors for runoff and soil erosion were different in the case of the mulched but not the PAM plots. Notably, the plots on the lower half of the slope registered bigger runoff and erosion figures than those on the upper half of the slope. This could be explained by differences in fire intensity and, ultimately, in pre-fire standing biomass.

  4. Reproducible protein analysis by CE using linear polyacrylamide-coated capillaries and hydrochloric acid rinsing.

    PubMed

    Suratman, Adhitasari; Wätzig, Hermann

    2007-07-01

    Hydrochloric acid was investigated as a rinsing reagent to remove adsorbed proteins from linear polyacrylamide-coated capillaries for electrophoresis. Three model proteins were used, namely cytochrome c as a basic protein, beta-lactoglobulin as an acidic protein, and beta-casein as a more easily denaturing protein. In order to regenerate capillary surfaces, they have been rinsed for 5 min with 2 M hydrochloric acid, 5 min with water, and then 30 min with buffer after every tenth run. It was found important to perform this regeneration procedure on time. The obtained results show good repeatability of the apparent EOF mobility with percentage RSDs below 3% (n = 60) in various cases. These good results were mainly confirmed in long-term series with more than 200 runs each. Only very high concentrations (175 microM) of beta-lactoglobulin and beta-casein at pH 3.5 gave RSD% values above 5%. For these conditions, the further test of 85% m/m phosphoric acid as rinsing reagent showed a good repeatability of the apparent EOF mobilities as well.

  5. Adsorption of cationic polyacrylamide at the cellulose-liquid interface: a neutron reflectometry study.

    PubMed

    Su, Jielong; Garvey, Christopher J; Holt, Stephen; Tabor, Rico F; Winther-Jensen, Bjorn; Batchelor, Warren; Garnier, Gil

    2015-06-15

    The layer thickness and density of high molecular weight cationic polyacrylamide (CPAM) adsorbed at the cellulose-water interface was quantified by neutron reflectometry. The thickness of a full monolayer of CPAM of constant molecular weight (13 MD) but different charge densities, adsorbed with or without NaCl (10(-3) M), was studied. Thin cellulose films (40±7 Å) of roughness <10 Å were produced by spin coating a cellulose acetate-acetone solution and regenerating by alkaline hydrolysis. Film smoothness was greatly improved by controlling the concentration of cellulose acetate (0.13 wt%) and the hydrolysis time in sodium methoxide. The adsorption thickness of CPAM (40% charge 13 MD) at the solid-D2O interface was 43±4 Å on cellulose and 13±2 Å on silicon, an order of magnitude smaller than the CPAM radius of gyration. At constant molecular weight, the thickness of the CPAM layer adsorbed on cellulose increases with polymer charge density (10±1 Å at 5%). Addition of 10(-3) M NaCl decreased the thickness of CPAM layer already adsorbed on cellulose. However, the adsorption layer on cellulose of a CPAM solution equilibrated in 10(-3) M NaCl is much thicker (89±11 Å for 40% CPAM). For high molecular weight CPAMs adsorbed from solution under constant conditions, the adsorption layer can be varied by 1 order of magnitude via control of the variables affecting electrostatic intra- and inter-polymer chain interactions.

  6. Adsorption of Hg2+ from aqueous solution onto polyacrylamide/attapulgite.

    PubMed

    Zhao, Yijiang; Chen, Yan; Li, Meisheng; Zhou, Shouyong; Xue, Ailian; Xing, Weihong

    2009-11-15

    Polyacrylamide/attapulgite (PAM/ATP) was prepared by the solution polymerization of acrylamide (AM) onto gamma-methacryloxypropyl trimethoxy silane (KH-570)-modified attapulgite (ATP). PAM/ATP was characterized using Fourier transform infrared (FTIR) and X-ray photoelectron spectroscopy (XPS). The effects of contact time, adsorbent dosage, and pH of the initial solution on the adsorption capacities for Hg(2+) were investigated. The adsorption process was rapid; 88% of adsorption occurred within 5 min and equilibrium was achieved at around 40 min. The equilibrium data fitted the Langmuir sorption isotherms well, and the maximum adsorption capacity of Hg(2+) onto PAM/ATP was found to be 192.5 mg g(-1). The adsorption kinetics of PAM/ATP fitted a pseudo-second-order kinetic model. Our results suggest that chemisorption processes could be the rate-limiting steps in the process of Hg(2+) adsorption. Hg(2+) adsorbed onto PAM/ATP could be effectively desorbed in hot acetic acid solution, and the adsorption capacity of the regenerated adsorbents could still be maintained at 95% by the sixth cycle.

  7. Enhancing the sorption capacity of CTMA-bentonite by simultaneous intercalation of cationic polyacrylamide.

    PubMed

    Wang, Tong; Zhu, Jianxi; Zhu, Runliang; Ge, Fei; Yuan, Peng; He, Hongping

    2010-06-15

    The saturated level of cationic exchange capacity (CEC) of bentonite by organic cations can significantly influence the sorption capacity of the resulting organobentonites. In this work cationic polyacrylamide (CPAM) was applied to saturate part of the CEC of the cetyltrimethylammonium (CTMA) modified bentonite, with the aim to enhance their sorption capacity. XRD was applied to investigate the basal spacings of the organobentonites with and without CPAM, and the sorption characteristics of the organobentonites towards phenol and nitrobenzene to CTMA-bentonite was also studied. The XRD characterization results showed that the resulting organobentonites (C/P-Bt) had larger basal spacings than the CTMA modified bentonite (C-Bt), which indicated that both CPAM and CTMA could intercalate into the interlayer spaces of C/P-Bt. The saturated CEC of C/P-Bt increased with the intercalated amounts of CPAM. The sorption coefficients (K(d)) of phenol and nitrobenzene on C/P-Bt were shown to first increase with the saturated CEC until the maximum, and then began to decrease as the saturated CEC further increased. The maximum increase of K(d) reached 41% and 23% for phenol and nitrobenzene, respectively, which could be ascribed to the enhanced affinity of the adsorbed CTMA aggregates towards the sorbates. Results of this work showed that the simultaneous intercalation of CPAM could be a novel method to enhance the sorption capacity of the traditional organobentonites.

  8. Response of hydroponically grown head lettuce on residual monomer from polyacrylamide.

    PubMed

    Mroczek, E; Konieczny, P; Kleiber, T; Waśkiewicz, A

    2014-01-01

    The aim was to assess acrylamide monomer (AMD) uptake by hydroponically grown lettuce. Lettuce was cultivated by applying plant tissue testing in a recycled system by the use of nutrient solutions prepared with two water-soluble flocculants F3 and F4 containing 176 and 763 mg kg(-1) of AMD, respectively. The effects on growth, fresh weight and plant leaf quality were evaluated by comparing these treatments and one control standard nutrient solution typically recommended for lettuce hydroponic cultivation. To assess the nutritional status of lettuce, samples were collected and lyophilised before determination of the selected micro- and macro-element contents. An HPLC with photodiode array detector method was applied to determine AMD in both selected flocculants and dried plant samples. Results show that lettuces cultivated under the conditions described above absorb AMD from nutrient solutions into their leaves. The AMD presence in recycled nutrient solutions has a negative influence on the growth of lettuce, reducing their average fresh weight and average number of leaves. The study confirmed that the problem of AMD mobility and its accumulation risk in plants should to be an important topic with respect to safe polyacrylamide (PAM) handling in the agro food area.

  9. Effect of residual monomer from polyacrylamide on head lettuce grown in peat substrate.

    PubMed

    Mroczek, Ewelina; Kleiber, Tomasz; Konieczny, Piotr; Waśkiewicz, Agnieszka

    2015-01-01

    The paper investigates the migration of the acrylamide monomer (AMD) to lettuce chosen as a test plant growing in an organic medium (peat substrate). Polyacrylamide (PAM)-based flocculant added to the growing medium contained no more than 1000 mg kg(-1) of AMD. Plants were grown with varied doses of PAM preparation (0.5-3.0 mg dm(-3) of peat substrate) to compare the results with the control sample. The determination of AMD content, chlorophyll content, weight of the lettuce head, and also analysis of macro- and micro-elements in lyophilised test material was made under the same analytical conditions. The results showed that lettuce plants absorb AMD to the leaves from the peat substrate. The AMD uptake has a negative impact on the growth of lettuce. It reduces the average fresh weight of heads and destabilises the mineral composition of the plant. Therefore, concern related to the transfer risk of the residual AMD from sludge used for organic fertilisation of edible plants still remains a crucial question from a food and consumer safety point of view. To ensure consumer safety, the fate of the AMD following the application of PAM to cropland should be carefully monitored in the whole food chain.

  10. Kinetics and thermodynamics of biodegradation of hydrolyzed polyacrylamide under anaerobic and aerobic conditions.

    PubMed

    Zhao, Lanmei; Bao, Mutai; Yan, Miao; Lu, Jinren

    2016-09-01

    Kinetics and thermodynamics of hydrolyzed polyacrylamide (HPAM) biodegradation in anaerobic and aerobic activated sludge biochemical treatment systems were explored to determine the maximum rate and feasibility of HPAM biodegradation. The optimal nutrient proportions for HPAM biodegradation were determined to be 0.08g·L(-1) C6H12O6, 1.00g·L(-1) NH4Cl, 0.36g·L(-1) NaH2PO4 and 3.00g·L(-1) K2HPO4 using response surface methodology (RSM). Based on the kinetics, the maximum HPAM biodegradation rates were 16.43385mg·L(-1)·d(-1) and 2.463mg·L(-1)·d(-1) in aerobic and anaerobic conditions, respectively. The activation energy (Ea) of the aerobic biodegradation was 48.9897kJ·mol(-1). Entropy changes (ΔS) of biochemical treatment system decreased from 216.21J·K(-1) to 2.39J·K(-1). Thermodynamic windows of opportunity for HPAM biodegradation were drawn. And it demonstrated HPAM was biodegraded into acetic acid and CO2 under laboratory conditions. Growth-process equations for functional bacteria anaerobically grown on polyacrylic acid were constructed and it confirmed electron equivalence between substrate and product.

  11. Flexible polyacrylamide substrata for the analysis of mechanical interactions at cell-substratum adhesions

    NASA Technical Reports Server (NTRS)

    Beningo, Karen A.; Lo, Chun-Min; Wang, Yu-Li

    2002-01-01

    We have described a powerful tool for the study of mechanical interactions between cells and their physical environment. Although the approach has already been used in a variety of ways to measure traction forces and to characterize active and passive responses of cultured cells to mechanical stimulation, it can be extended easily and combined with other microscopic approaches, including fluorescent analog imaging (Beningo et al., 2001), photobleaching, calcium imaging, micromanipulation, and electrophysiology. This method will be particularly useful for studying the functions of various components at focal adhesions, and the effects of mechanical forces on focal adhesion-mediated signal transduction. In addition, the method can be extended to a 3D setting, e.g., by sandwiching cultured cells between two layers of polyacrylamide to create an environment mimicking that in the tissue of a multicellular organism. Whereas chemical interactions between cells and the environment have been investigated extensively, many important questions remain as to the role of physical forces in cellular functions and the interplay between chemical and physical mechanisms of communication. The present approach, as well as other approaches capable of probing physical interactions, should fill in this important gap in the near future.

  12. Hydrolyzed polyacrylamide grafted maize starch based microbeads: application in pH responsive drug delivery.

    PubMed

    Setty, C Mallikarjuna; Deshmukh, Anand S; Badiger, Aravind M

    2014-09-01

    The present study details the synthesis, characterization and pharmaceutical application of hydrolysed polyacrylamide grafted maize starch (HPam-g-MS) as promising polymeric material for the development of pH responsive microbeads. Different grades of graft copolymer were synthesized by changing the net microwave irradiation time, while keeping all other factors constant. Acute oral toxicity study performed in rodents ensured the bio-safety of graft copolymer for clinical application. Various batches of aceclofenac loaded microbeads were prepared by ionic gelation method using synthesized graft copolymers and evaluated for formulation parameters. FTIR spectroscopy confirmed the chemical compatibility between drug and graft copolymer. Results of in vitro release study (USP type-II) carried out in two different pH media (pH 1.2 acid buffer and pH 7.4 phosphate buffer) showed that release rate of drug from developed microbeads was a function of both: (a) surrounding pH and (b) the matrix composition. The drug release was relatively higher at alkaline pH as compared to acidic pH and this feature is desirable from viewpoint of site specific drug delivery. A direct correlation was observed between percentage grafting and microbeads performance and it presents a scope for further research on application and optimization of HPam-g-MS based microbeads as drug delivery carriers.

  13. Toxicity of anionic polyacrylamide formulations when used for erosion control in agriculture.

    PubMed

    Weston, Donald P; Lentz, Rodrick D; Cahn, Michael D; Ogle, R Scott; Rothert, Amanda K; Lydy, Michael J

    2009-01-01

    Addition of anionic polyacrylamide (PAM) to agricultural irrigation water can dramatically reduce erosion of soils. However, the toxicity of PAM to aquatic life, while often claimed to be low, has not been thoroughly evaluated. Five PAM formulations, including two oil-based products, one water-based product, one granular product and one tablet product, were evaluated for acute and/or chronic toxicity to five species commonly used for freshwater toxicity testing [Hyalella azteca (Saussure), Chironomus dilutus (Shobanov et al.), Ceriodaphnia dubia (Richard), Pimephales promelas (Rafinesque), and Selenastrum capricornutum (Printz)]. When applied as an oil-based product, acute toxicity was seen to four of the five species at concentrations less than the 10 mg/L that is often used for erosion control. Toxicity was diminished, but still remained, after passage of the irrigation water across an agricultural field, indicating a potential impact to nearby surface waters. Results from the non-oil-based products indicated minimal toxicity associated with PAM even at concentrations 10 times those used in agriculture when applied in the granular form, as a tablet, or in a water-based liquid. These data suggest that other agents in the oil-based products, such as surfactants or emulsifiers, rather than the PAM itself, contribute to the toxicity. Care is required in selecting an appropriate PAM formulation when the potential exists for entry of tailwater to nearby surface waters.

  14. Harvesting Chlorella vulgaris by magnetic flocculation using Fe₃O₄ coating with polyaluminium chloride and polyacrylamide.

    PubMed

    Zhao, Yuan; Liang, Wenyan; Liu, Lijun; Li, Feizhen; Fan, Qianlong; Sun, Xiaoli

    2015-12-01

    The harvesting of Chlorella vulgaris was investigated by magnetic flocculation, where the natural magnetite was used as magnetic seeds and the polyaluminium chloride (PACl) and polyacrylamide (PAM) were used as the coating polymer on the Fe3O4 surface. The composite modes of PACl, PAM, and Fe3O4 and their effects on harvesting were studied. The results showed that adding the composite PACl/Fe3O4 first (at (0.625 mmol Al/L)/(10 g/L)) followed by the addition PAM (at 3mg/L) was the optimum dosing strategy. Following this strategy, 99% of cells could be harvested in less than 0.5 min, and it could overcome negative impacts from pH and algal organic matter. Compared to PACl, ζ-potentials of PACl/Fe3O4 were found to be increased substantially from -4.9-8.5 mV to 1.5-19.5 mV at pH range 2.1-12.3. The charge neutralization of PACl/Fe3O4 and sweeping of PAM play an important role in magnetic harvesting of microalgal cells.

  15. Methylene blue covalently loaded polyacrylamide nanoparticles for enhanced tumor-targeted photodynamic therapy†

    PubMed Central

    Qin, Ming; Hah, Hoe Jin; Kim, Gwangseong; Nie, Guochao; Lee, Yong-Eun Koo

    2013-01-01

    The use of targeted nanoparticles (NPs) as a platform for loading photosensitizers enables selective accumulation of the photosensitizers in the tumor area, while maintaining their photodynamic therapy (PDT) effectiveness. Here two novel kinds of methylene blue (MB)-conjugated polyacrylamide (PAA) nanoparticles, MBI-PAA NPs and MBII-PAA NPs, based on two separate MB derivatives, are developed for PDT. This covalent conjugation with the NPs (i) improves the loading of MB, (ii) prevents any leaching of MB from the NPs and (iii) protects the MB from the effects of enzymes in the biological environment. The loading of MB into these two kinds of NPs was controlled by the input amount, resulting in concentrations with optimal singlet oxygen production. For each of the MB-NPs, the highest singlet oxygen production was found for an MB loading of around 11 nmol mg−1. After attachment of F3 peptide groups, for targeting, each of these NPs was taken up, selectively, by MDA-MB-435 tumor cells, in vitro. PDT tests demonstrated that both kinds of targeted NPs resulted in effective tumor cell kill, following illumination, while not causing dark toxicity. PMID:21479315

  16. Transfer and degradation of polyacrylamide-based flocculants in hydrosystems: a review.

    PubMed

    Guezennec, A G; Michel, C; Bru, K; Touze, S; Desroche, N; Mnif, I; Motelica-Heino, M

    2015-05-01

    The aim of this review was to summarize information and scientific data from the literature dedicated to the fate of polyacrylamide (PAM)-based flocculants in hydrosystems. Flocculants, usually composed of PAMs, are widely used in several industrial fields, particularly in minerals extraction, to enhance solid/liquid separation in water containing suspended matter. These polymers can contain residual monomer of acrylamide (AMD), which is known to be a toxic compound. This review focuses on the mechanisms of transfer and degradation, which can affect both PAM and residual AMD, with a special attention given to the potential release of AMD during PAM degradation. Due to the ability of PAM to adsorb onto mineral particles, its transport in surface water, groundwater, and soils is rather limited and restricted to specific conditions. PAM can also be a subject of biodegradation, photodegradation, and mechanical degradation, but most of the studies report slow degradation rates without AMD release. On the contrary, the adsorption of AMD onto particles is very low, which could favor its transfer in surface waters and groundwater. However, AMD transfer is likely to be limited by quick microbial degradation.

  17. Polyacrylamide-metal nanocomposites: one-pot synthesis, antibacterial properties, and thermal stability

    NASA Astrophysics Data System (ADS)

    Li, Cuiyan; Cai, Yanling; Zhu, Yihua; Ma, Mingguo; Zheng, Wei; Zhu, Jiefang

    2013-09-01

    The incorporation of inorganic nanoparticles into polymers is a hot research spot, since it endows the nanocomposites with new or improved properties by exploiting synergistic effects. Here we report a facile one-pot synthesis of polyacrylamide (PAM)-metal (M = Au, Ag, or Pd) nanocomposites in ethylene glycol (EG). The simultaneous polymerization of the acylamide (AM) monomer and formation of metal nanoparticles lead to a homogeneous distribution of metal nanoparticles in the PAM matrix. The sizes of Au, Ag, and Pd nanoparticles are 55.50 ± 10.6, 14.15 ± 2.57, and 7.74 ± 1.82 nm, respectively. The reaction system only includes EG, AM monomer, and corresponding metal salt. EG acts as both the solvent and the reducing reagent. Also, no initiator for AM polymerization and no surfactant for stabilization of metal nanoparticles are used. Furthermore, this simple synthetic route does not rely on any special or expensive equipment, thus can be exploited to the synthesis of similar polymer-inorganic nanocomposites. Compared to PAM, the PAM-metal nanocomposites showed enhanced thermal stability and antibacterial properties.

  18. Competition between kaolinite flocculation and stabilization in divalent cation solutions dosed with anionic polyacrylamides.

    PubMed

    Lee, Byung Joon; Schlautman, Mark A; Toorman, Erik; Fettweis, Michael

    2012-11-01

    Divalent cations have been reported to develop bridges between anionic polyelectrolytes and negatively-charged colloidal particles, thereby enhancing particle flocculation. However, results from this study of kaolinite suspensions dosed with various anionic polyacrylamides (PAMs) reveal that Ca(2+) and Mg(2+) can lead to colloid stabilization under some conditions. To explain the opposite but coexisting processes of flocculation and stabilization with divalent cations, a conceptual flocculation model with (1) particle-binding divalent cationic bridges between PAM molecules and kaolinite particles and (2) polymer-binding divalent cationic bridges between PAM molecules is proposed. The particle-binding bridges enhanced flocculation and aggregated kaolinite particles in large, easily-settleable flocs whereas the polymer-binding bridges increased steric stabilization by developing polymer layers covering the kaolinite surface. Both the particle-binding and polymer-binding divalent cationic bridges coexist in anionic PAM- and kaolinite-containing suspensions and thus induce the counteracting processes of particle flocculation and stabilization. Therefore, anionic polyelectrolytes in divalent cation-enriched aqueous solutions can sometimes lead to the stabilization of colloidal particles due to the polymer-binding divalent cationic bridges.

  19. Arsenic remediation from drinking water by synthesized nano-alumina dispersed in chitosan-grafted polyacrylamide.

    PubMed

    Saha, Suparna; Sarkar, Priyabrata

    2012-08-15

    An arsenic adsorbent comprising alumina nanoparticles dispersed in polymer matrix was developed and its removal potential studied. Alumina nanoparticles were prepared by reverse microemulsion technique and these were immobilized on chitosan grafted polyacrylamide matrix by in situ dispersion. The loading capacity of this new synthesized adsorbent was found to be high (6.56 mg/g). Batch adsorption studies were performed as a function of contact time, initial arsenic concentration, pH and presence of competing anions. The removal was found to be pH dependent, and maximum removal was obtained at pH 7.2 while the equilibrium time was 6h. The equilibrium adsorption data fitted very well with Freundlich isotherm. However, the D-R isotherm studies indicated that chemisorptions might play an important role. This was also confirmed by the FTIR study of the arsenic loaded adsorbent. A mechanism of arsenic sorption by the new polymeric adsorbent has been proposed. The regeneration study of the adsorbent resulted in retention of 94% capacity in the fifth cycle. An optimum pH of 7.2, operation at normal temperature, high adsorption capacity and good recycle potential of this new adsorbent would make it an ideal material for removal of arsenic from drinking water.

  20. Nanocrystal/sol-gel nanocomposites

    DOEpatents

    Petruska, Melissa A.; Klimov, Victor L.

    2007-06-05

    The present invention is directed to solid composites including colloidal nanocrystals within a sol-gel host or matrix and to processes of forming such solid composites. The present invention is further directed to alcohol soluble colloidal nanocrystals useful in formation of sol-gel based solid composites.

  1. Crystallization of steroids in gels

    NASA Astrophysics Data System (ADS)

    Kalkura, S. Narayana; Devanarayanan, S.

    1991-03-01

    The crystal growth and characterization of certain steriods, viz., cholesterol, cholesteryl acetate, β-sitosterol, progesterone and testosterone, in a silica gel medium is discussed. The present study shows that the single test tube diffusion method can be used to grow crystals of steroids in a silica gel medium by the reduction of steroid solubility.

  2. Nanocrystal/sol-gel nanocomposites

    SciTech Connect

    Petruska, Melissa A; Klimov, Victor L

    2012-06-12

    The present invention is directed to solid composites including colloidal nanocrystals within a sol-gel host or matrix and to processes of forming such solid composites. The present invention is further directed to alcohol soluble colloidal nanocrystals useful in formation of sol-gel based solid composites

  3. A new agarose gel model

    SciTech Connect

    Hasenfeld, A.; Pepke, E.; Lim, H.A.; Cantor, C.R.

    1993-12-31

    A new agarose gel model is introduced, which corresponds to what the authors believe agarose gels look like microscopically. While the scientific literature is filled with studies of the microscopic structure of agarose, the fact remains that there is no unambiguous and exact model of its underlying structure. Given this, the authors are left to construct their own model numerically.

  4. Living bacteria in silica gels

    NASA Astrophysics Data System (ADS)

    Nassif, Nadine; Bouvet, Odile; Noelle Rager, Marie; Roux, Cécile; Coradin, Thibaud; Livage, Jacques

    2002-09-01

    The encapsulation of enzymes within silica gels has been extensively studied during the past decade for the design of biosensors and bioreactors. Yeast spores and bacteria have also been recently immobilized within silica gels where they retain their enzymatic activity, but the problem of the long-term viability of whole cells in an inorganic matrix has never been fully addressed. It is a real challenge for the development of sol-gel processes. Generic tests have been performed to check the viability of Escherichia coli bacteria in silica gels. Surprisingly, more bacteria remain culturable in the gel than in an aqueous suspension. The metabolic activity of the bacteria towards glycolysis decreases slowly, but half of the bacteria are still viable after one month. When confined within a mineral environment, bacteria do not form colonies. The exchange of chemical signals between isolated bacteria rather than aggregates can then be studied, a point that could be very important for 'quorum sensing'.

  5. High-resolution two-dimensional polyacrylamide gel electrophoresis reveals a glucose-response protein of 65 kDa in pancreatic islet cells

    SciTech Connect

    Collins, H.W.; Buettger, C.; Matschinsky, F. )

    1990-07-01

    High-resolution two-dimensional PAGE was used to search for glucose-response proteins in isolated pancreatic islets that were labeled with ({sup 35}S)methionine at ambient glucose concentrations of 0-18 mM. A 65-kDa protein, isoelectric focusing point of approximately 6.6-7.0, was discovered that showed at least a 20-fold stimulation of radiolabeling when glucose in the labeling medium was increased from 3 to 18 mM, in contrast to a 2.5-fold enhancement of label incorporation into total islet proteins. This 65-kDa protein is evident after 30 min of labeling with 18 mM glucose and is preferentially synthesized compared to its nearest neighbors after both 30 and 60 min of labeling. Glucose induction of the 65-kDa protein was virtually blocked by D-mannoheptulose. Glucose induction of this 65-kDa protein is in practically all aspects comparable to glucose induction of insulin and glucokinase in pancreatic beta cells. A working hypothesis is developed proposing that glucose-response proteins or glucospondins are pivotal constituents of pancreatic islet cells and that their discovery and exploration promise new insights into normal and pathological islet cell function.

  6. Development of a Low-cost and High-throughput Polyacrylamide Gel System for Peanut Genotyping with Simple Sequence Repeat (SSR) Markers.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Traditionally, peanut cultivar development has been dominated by conventional breeding methods, which have greatly increased yield and will continue to play an important role in peanut genetic improvement. Applications of MAS (marker-assisted selection) in plant breeding have been shown to increase ...

  7. Development of a Low-cost and High-throughput Polyacrylamide Gel System for Peanut Genotyping with Simple Sequence repeat (SSR) Markers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Traditionally, peanut cultivar development has been dominated by conventional breeding methods, which have greatly increased yield and will continue to play an important role in peanut genetic improvement. Applications of MAS (marker-assisted selection) in plant breeding have been shown to increase ...

  8. Application of NMR spectroscopy and multidimensional imaging to the gelcasting process and in-situ real-time monitoring of cross-linking polyacrylamide gels

    SciTech Connect

    Ahuja, S.; Dieckman, S.L.; Gopalsami, N.

    1995-04-01

    In the gelcasting process, a slurry of ceramic powder in a solution of organic monomers is cast in a mold. The process is different from injection molding in that it separates mold-filling from setting during conversion of the ceramic slurry to a formed green part. In this work, NMR spectroscopy and imaging have been conducted for in-situ monitoring of the gelation process and for mapping the polymerization. {sup 1}H nuclear magnetic resonance spectra have been obtained during polymerization of a premix of soluble reactive methacrylamide (monomer) and N, N`-methylene bisacrylamide (cross-linking molecules). The premix was polymerized by adding ammonium persulfate (initiator) and tetramethyl-ethylene-diamine (accelerator) to form long-chain, cross-linked polymers. The time-varying spin-lattice relaxation times T{sub 1} during polymerization have been studied at 25 and 35{degrees}C, and the variation of spectra and T{sub 1} with respect to extent of polymerization has been determined. To verify homogeneous polymerization, multidimensional NMR imaging was utilized for in-situ monitoring of the process. The intensities from the images are modeled and the correspondence shows a direct extraction of T{sub 1} data from the images.

  9. Qualitative and quantitative determination of R-phycoerythrin from Halymenia floresia (Clemente) C. Agardh by polyacrylamide gel using electrophoretic elution technique.

    PubMed

    Malairaj, Sathuvan; Muthu, Sakthivel; Gopal, Venkatesh Babu; Perumal, Palani; Ramasamy, Rengasamy

    2016-07-08

    R-Phycoerythrin is one of the phycobiliproteins widely found in seaweeds. In this study, we have shown to extract and purify R-Phycoerythrin from the south east cost Indian red seaweed Halymenia floresia. R-Phycoerythrin was extracted in 50mM phosphate buffer (pH 7.0). The preparative native PAGE purification was employed alternative to the chromatography and therefore can be scaled up efficiently. Both the yield and the purity of R-Phycoerythrin are very effective. The purified R-Phycoerythrin showed a single band on the examination by native PAGE electrophrosis. SDS-PAGE analysis showed five bands at 16kDa, 21kDa, 30kDa, 39kDa and 47kDa which corresponds to the α, β and γ', γ and αβ subunits. This preparative method for R-Phycoerythrin purification can offer a reference for R-Phycoerythrin purification from other marine red macro algae.

  10. Analysis of photoaffinity-labeled aryl hydrocarbon receptor heterogeneity by two-dimensional gel electrophoresis

    SciTech Connect

    Perdew, G.H.; Hollenback, C.E. )

    1990-07-03

    The level of charge heterogeneity in the aryl hydrocarbon receptor (AhR) was examined by high-resolution denaturing two-dimensional (2D) gel electrophoresis. Hepa 1c1c7 cell cytosolic fraction was photoaffinity-labeled with 2-azido-3-({sup 125}I)-iodo-7,8-dibromodibenzo-p-dioxin and applied to isoelectric focusing (IEF) tube gels. After optimization of focusing conditions a broad peak of radioactivity was detected in the apparent pI range of 5.2-5.7. IEF tube gels were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by visualization of the radiolabeled AhR by autoradiography; three distinct isoforms were detected. The same 2D electrophoretic isoform pattern was obtained when the AhR from Hepa 1c1c7 was photoaffinity-labeled in cell culture. BP{sup r}Cl cells, a mutant line derived from Hepa 1c1c7 cells, contain an AhR that is unable to bind to DNA. Photoaffinity-labeled BP{sup r}Cl cytosolic fractions were subjected to 2D gel electrophoretic analysis resulting in essentially the same molecular weight and isoform pattern as seen in Hepa 1c1c7 cytosol. This result would suggest that if a mutation is present in the BP{sup r}Cl AhR it has not caused a significant change in its IEF pattern, although a small shift in the pI values was observed. Two-dimensional gel electrophoresis of photoaffinity-labeled cytosolic fractions from HeLa cells, the rat liver tumor cell line McA-RH777, and buffalo rat thymus revealed three isoforms, essentially the same isoform pattern as in Hepa 1c1c7 cells. This would indicate that despite the considerable molecular weight polymorphism between species the level of charge heterogeneity is high conserved.

  11. Automatic DNA Diagnosis for 1D Gel Electrophoresis Images using Bio-image Processing Technique

    PubMed Central

    2015-01-01

    Background DNA gel electrophoresis is a molecular biology technique for separating different sizes of DNA fragments. Applications of DNA gel electrophoresis include DNA fingerprinting (genetic diagnosis), size estimation of DNA, and DNA separation for Southern blotting. Accurate interpretation of DNA banding patterns from electrophoretic images can be laborious and error prone when a large number of bands are interrogated manually. Although many bio-imaging techniques have been proposed, none of them can fully automate the typing of DNA owing to the complexities of migration patterns typically obtained. Results We developed an image-processing tool that automatically calls genotypes from DNA gel electrophoresis images. The image processing workflow comprises three main steps: 1) lane segmentation, 2) extraction of DNA bands and 3) band genotyping classification. The tool was originally intended to facilitate large-scale genotyping analysis of sugarcane cultivars. We tested the proposed tool on 10 gel images (433 cultivars) obtained from polyacrylamide gel electrophoresis (PAGE) of PCR amplicons for detecting intron length polymorphisms (ILP) on one locus of the sugarcanes. These gel images demonstrated many challenges in automated lane/band segmentation in image processing including lane distortion, band deformity, high degree of noise in the background, and bands that are very close together (doublets). Using the proposed bio-imaging workflow, lanes and DNA bands contained within are properly segmented, even for adjacent bands with aberrant migration that cannot be separated by conventional techniques. The software, called GELect, automatically performs genotype calling on each lane by comparing with an all-banding reference, which was created by clustering the existing bands into the non-redundant set of reference bands. The automated genotype calling results were verified by independent manual typing by molecular biologists. Conclusions This work presents an

  12. A 2-dimensional MHD code & survey of the ``buckling'' phenomenon in cylindrical magnetic flux compression experiments

    NASA Astrophysics Data System (ADS)

    Xiao, Bo; Wang, Ganghua; Gu, Zhuowei; Computational Physics Team

    2015-11-01

    We made a 2-dimensional magneto-hydrodynamics Lagrangian code. The code handles two kinds of magnetic configuration, a (x-y) plane with z-direction magnetic field Bz and a (r-z) plane with θ-direction magnetic field Bθ. The solving of the MHD equations is split into a pure dynamical step (i.e., ideal MHD) and a diffusion step. In the diffusion step, the Joule heat is calculated with a numerical scheme based on an specific form of the Joule heat production equation, ∂eJ/∂t = ∇ . (η/μ0 º × (∇ × º)) -∂/∂t (1/2μ0 B2) , where the term ∂/∂t (1/2μ0 B2) is the magnetic field energy variation caused solely by diffusion. This scheme insures the equality of the total Joule heat produced and the total electromagnetic energy lost in the system. Material elastoplasticity is considered in the code. An external circuit is coupled to the magneto-hydrodynamics and a detonation module is also added to enhance the code's ability for simulating magnetically-driven compression experiments. As a first application, the code was utilized to simulate a cylindrical magnetic flux compression experiment. The origin of the ``buckling'' phenomenon observed in the experiment is explored.

  13. Data assimilation technique of 2-dimensional vertical temperature transport model (Case study: Tropical Pacific Ocean)

    NASA Astrophysics Data System (ADS)

    Nurfitri, Suliskania; Putri, Mutiara Rachmat

    2015-09-01

    Data assimilation technique was applied into 2-dimensional vertical temperature transport model to improve temperature results especially at thermocline layer that have large change toward depth. The simple case experiment only applies on baroclinic condition in Tropical Pacific Ocean (2°N and 137°E - 140°W). Model simulation was running for 2 years from January 1st 2011 to December 31st 2012 from surface to 500 m depth and verified to observation data from TAO (Tropical Atmosphere Ocean) at 3 locations, 147°E, 165°E, and 170°W. Data assimilation procedure which applied at 3 locations (156°E, 180°W, and 155°W) using Cressman analysis technique can reduce model's RMSE (Root Mean Square Error) toward depth until 55,7% and toward time until 64,5%. The largest error of model was found at 200 m depth, while the smallest found at the surface and 500 m depth.

  14. Role of surface defects on the formation of the 2-dimensional electron gas at polar interfaces

    NASA Astrophysics Data System (ADS)

    Artacho, Emilio; Aguado-Puente, Pablo

    2014-03-01

    The discovery of a 2-dimensional electron gas (2DEG) at the interface between two insulators, LaAlO3 and SrTiO3, has fuelled a great research activity on this and similar systems in the last years. The electronic reconstruction model, typically invoked to explain the formation of the 2DEG, while being intuitive and successful on predicting fundamental aspects of this phenomenon like the critical thickness of LaAlO3, fails to explain many other experimental observations. Oxygen vacancies, on the other hand, are known to dramatically affect the physical behaviour of this system, but their role at the atomic level is far from well understood. Here we perform ab initio simulations in order to assess whether the formation of oxygen vacancies at the surface of the polar material can account for various recent experimental results that defy the current theoretical understanding of these interfaces. We simulate SrTiO3/LaAlO3 slabs with various concentrations of surface oxygen vacancies and analyze the role of the defects on the formation of the metallic interface, their electrostatic coupling with the 2DEG and the interplay with the different instabilities of the materials involved. Financial support from Spanish MINECO under grant FIS2012-37549-C05-01. Computational resources provided by the Red Espñola de Supercomputación and DIPC.

  15. Substrate induced changes in atomically thin 2-dimensional semiconductors: Fundamentals, engineering, and applications

    NASA Astrophysics Data System (ADS)

    Sun, Yinghui; Wang, Rongming; Liu, Kai

    2017-03-01

    Substrate has great influences on materials syntheses, properties, and applications. The influences are particularly crucial for atomically thin 2-dimensional (2D) semiconductors. Their thicknesses are less than 1 nm; however, the lateral sizes can reach up to several inches or more. Therefore, these materials must be placed onto a variety of substrates before subsequent post-processing techniques for final electronic or optoelectronic devices. Recent studies reveal that substrates have been employed as ways to modulate the optical, electrical, mechanical, and chemical properties of 2D semiconductors. In this review, we summarize recent progress upon the effects of substrates on properties of 2D semiconductors, mostly focused on 2D transition metal dichalcogenides, through viewpoints of both fundamental physics and device applications. First, we discuss various effects of substrates, including interface strain, charge transfer, dielectric screening, and optical interference. Second, we show the modulation of 2D semiconductors by substrate engineering, including novel substrates (patterned substrates, 2D-material substrates, etc.) and active substrates (phase transition materials, ferroelectric materials, flexible substrates, etc.). Last, we present prospectives and challenges in this research field. This review provides a comprehensive understanding of the substrate effects, and may inspire new ideas of novel 2D devices based on substrate engineering.

  16. Carbonic anhydrase-facilitated CO2 absorption with polyacrylamide buffering bead capture

    SciTech Connect

    Dilmore, Robert; Griffith, Craid; Liu, Zhu; Soong, Yee; Hedges, Sheila W.; Koepsel, Richard; Ataai, M

    2009-07-01

    A novel CO2 separation concept is described wherein the enzyme carbonic anhydrase (CA) is used to increase the overall rate Of CO2 absorption after which hydrated CO2 reacts with regenerable amine-bearing polyacrylamide buffering beads (PABB). Following saturation of the material's immobilized tertiary amines, CA-bearing carrier water is separated and recycled to the absorption stage while CO2-loaded material is thermally regenerated. Process application of this concept would involve operation of two or more columns in parallel with thermal regeneration with low-pressure steam taking place after the capacity of a column of amine-bearing polymeric material was exceeded. PABB CO2- bearing capacity was evaluated by thermogravimetric analysis (TGA) for beads of three acrylamido buffering monomer ingredient concentrations: 0 mol/kg bead, 0.857 mol/kg bead, and 2 mol/kg bead. TGA results demonstrate that CO2- bearing capacity increases with increasing PABB buffering concentration and that up to 78% of the theoretical CO2- bearing capacity was realized in prepared PABB samples (0.857 mol/kg recipe). The highest observed CO2-bearing capacity of PABB was 1.37 mol of CO2 per kg dry bead. TGA was also used to assess the regenerability Of CO2-loaded PABB. Preliminary results suggest that CO2 is partially driven from PABB samples at temperatures as low as 55 degrees C, with complete regeneration occurring at 100 degrees C. Other physical characteristics of PABB are discussed. In addition, the effectiveness of bovine carbonic anhydrase for the catalysis Of CO2 dissolution is evaluated. Potential benefits and drawbacks of the proposed process are discussed. Published by Elsevier Ltd.

  17. Ti 3 C 2 T x (MXene)–polyacrylamide nanocomposite films

    DOE PAGES

    Naguib, Michael; Saito, Tomonori; Lai, Sophia; ...

    2016-07-20

    Polymer nanocomposite films are of great interest due to their enhanced properties over base polymers. By incorporating 2D titanium carbide a representative of a new family of 2D materials, MXenes, as nanofillers into a water soluble polyacrylamide (PAM) matrix, the resulting films benefit from the flexibility, robustness, and processability of PAM, as well as the conductivity and mechanical properties of MXene fillers. We report on manufacturing and characterization of MXene-PAM nanocomposite films. Dimethylsulfoxide (DMSO) intercalation in-between the Ti3C2-based MXene layers led to full delamination of the MXene layers and hence a uniform dispersion of hydrophilic MXene nanosheets in aqueous PAMmore » solutions was achieved. Moreover, the polymer composite solutions of up to 75 wt.% MXene loading were sonicated and cast onto large Teflon trays and dried at room temperature to produce shiny black films. The observation of reduced 0002 peaks of Ti3C2Tx phase in X-ray diffraction patterns and TEM images indicate the presence of well dispersed nanoflakes. The as-prepared composite films are flexible and the conductivity was increased significantly to 3 x 10-3 S cm-1 for 6 wt. % MXene-PAM films. With high MXene loading, some non-uniformity between the top and bottom surfaces was observed. This could be due to the segregation of MXene layers in composite films during drying. Finally, the power law dependence of conductivity above the percolation threshold is presented through detailed conductivity measurements.« less

  18. Coagulation of soil suspensions containing nonionic or anionic surfactants using chitosan, polyacrylamide, and polyaluminium chloride.

    PubMed

    Chatterjee, Tania; Chatterjee, Sudipta; Lee, Dae S; Lee, Min W; Woo, Seung Han

    2009-06-01

    Effective coagulation and separation of particles in a soil-washed solution is required for a successful soil washing process. The effectiveness of chitosan (CS), a polycationic biodegradable polymer, as a coagulant was compared to polyacrylamide (PAA) and polyaluminium chloride (PAC) for the coagulation of a soil suspension (5 gL(-1)). The effect of surfactants in the coagulation process was investigated using Triton X-100 (TX-100), a nonionic surfactant, and sodium dodecyl sulfate (SDS), an anionic surfactant. CS (5 mgL(-1)) removed 86% and 63% of the suspended soil in the presence of TX-100 (5 gL(-1)) and SDS (5 gL(-1)), respectively, after 30 min at a pH of 6. The results prove that coagulation in the presence of TX-100 is more effective than with SDS. CS was found to be more efficient compared to PAA and PAC under all coagulation conditions. The optimum concentration of CS required for maximum coagulation of soil suspension was 5 mgL(-1). PAA and PAC could not achieve the same degree soil removal as CS even after increasing their concentrations up to 50 mgL(-1). Maximum levels of 50% and 60% soil removal were achieved using PAA (50 mgL(-1)) and PAC (50 mgL(-1)), respectively, after 30 min from a 5 gL(-1) suspension containing TX-100 (5 gL(-1)). The soil coagulation process was found to decrease with an increase in the pH of the suspension, and maximum coagulation was achieved with an acidic pH.

  19. Mechanical Failure in Colloidal Gels

    NASA Astrophysics Data System (ADS)

    Kodger, Thomas Edward

    When colloidal particles in a dispersion are made attractive, they aggregate into fractal clusters which grow to form a space-spanning network, or gel, even at low volume fractions. These gels are crucial to the rheological behavior of many personal care, food products and dispersion-based paints. The mechanical stability of these products relies on the stability of the colloidal gel network which acts as a scaffold to provide these products with desired mechanical properties and to prevent gravitational sedimentation of the dispersed components. Understanding the mechanical stability of such colloidal gels is thus of crucial importance to predict and control the properties of many soft solids. Once a colloidal gel forms, the heterogeneous structure bonded through weak physical interactions, is immediately subject to body forces, such as gravity, surface forces, such as adhesion to a container walls and shear forces; the interplay of these forces acting on the gel determines its stability. Even in the absence of external stresses, colloidal gels undergo internal rearrangements within the network that may cause the network structure to evolve gradually, in processes known as aging or coarsening or fail catastrophically, in a mechanical instability known as syneresis. Studying gel stability in the laboratory requires model colloidal system which may be tuned to eliminate these body or endogenous forces systematically. Using existing chemistry, I developed several systems to study delayed yielding by eliminating gravitational stresses through density matching and cyclic heating to induce attraction; and to study syneresis by eliminating adhesion to the container walls, altering the contact forces between colloids, and again, inducing gelation through heating. These results elucidate the varied yet concomitant mechanisms by which colloidal gels may locally or globally yield, but then reform due to the nature of the physical, or non-covalent, interactions which form

  20. Adhesive, elastomeric gel impregnating composition

    DOEpatents

    Shaw, David Glenn; Pollard, John Randolph; Brooks, Robert Aubrey

    2002-01-01

    An improved capacitor roll with alternating film and foil layers is impregnated with an adhesive, elastomeric gel composition. The gel composition is a blend of a plasticizer, a polyol, a maleic anhydride that reacts with the polyol to form a polyester, and a catalyst for the reaction. The impregnant composition is introduced to the film and foil layers while still in a liquid form and then pressure is applied to aid with impregnation. The impregnant composition is cured to form the adhesive, elastomeric gel. Pressure is maintained during curing.

  1. Polyoxometalate-based Supramolecular Gel

    PubMed Central

    He, Peilei; Xu, Biao; Liu, Huiling; He, Su; Saleem, Faisal; Wang, Xun

    2013-01-01

    Self-assemblyings of surfactant-encapsulated Wells-Dawson polyoxometalates (SEPs) nanobuilding blocks in butanone and esters yielded supramolecular gels showing thermo and photo responsive properties. The gels can be further polymerized if unsaturated esters were used and subsequently electrospinned into nanowires and non-woven mats. The as-prepared non-woven mats have a Young's modulus as high as 542.55 MPa. It is believed that this supramolecular gel is a good platform for polyoxometalates processing. PMID:23666013

  2. Differentiation between fresh and frozen-thawed sea bass (Dicentrarchus labrax) fillets using two-dimensional gel electrophoresis.

    PubMed

    Ethuin, Pierrette; Marlard, Sylvain; Delosière, Mylène; Carapito, Christine; Delalande, François; Van Dorsselaer, Alain; Dehaut, Alexandre; Lencel, Valérie; Duflos, Guillaume; Grard, Thierry

    2015-06-01

    This study aimed to identify a protein marker that can differentiate between fresh skinless and frozen-thawed sea bass (Dicentrarchus labrax) fillets using the two-dimensional polyacrylamide gel electrophoresis (2-DE) technique. Distinct gel patterns, due to proteins with low molecular weight and low isoelectric points, distinguished fresh fillets from frozen-thawed ones. Frozen-thawed fillets showed two specific protein spots as early as the first day of the study. However, these spots were not observed in fresh fillets until at least 13days of storage between 0 and 4°C, fillets were judged, beyond this period, fish were unfit for human consumption as revealed by complementary studies on fish spoilage indicators namely total volatile basic nitrogen and biogenic amines. Mass spectrometry identified the specific proteins as parvalbumin isoforms. Parvalbumins may thus be useful markers of differentiation between fresh and frozen-thawed sea bass fillets.

  3. Titanium Dioxide Photocatalytic Polymerization of Acrylamide for Gel Electrophoresis (TIPPAGE) of Proteins and Structural Identification by Mass Spectrometry.

    PubMed

    Zhang, Wenyang; Yuan, Zhiwei; Huang, Lulu; Kang, Jie; Jiang, Ruowei; Zhong, Hongying

    2016-02-11

    Polyacrylamide gel electrophoresis (PAGE) coupled with mass spectrometry has been well established for separating, identifying and quantifying protein mixtures from cell lines, tissues or other biological samples. The copolymerization process of acrylamide and bis-acrylamide is the key to mastering this powerful technique. In general, this is a vinyl addition reaction initiated by free radical-generating reagents such as ammonium persulfate (APS) and tetramethylethylenediamine (TEMED) under basic pH and degassing experimental condition. We report herein a photocatalytic polymerization approach that is based on photo-generated hydroxyl radicals with nanoparticles of titanium dioxide. It was shown that the polymerization process is greatly accelerated in acidic condition when ultraviolet light shots on the gel solution containing TiO2 nanoparticles without degassing. This feature makes it very useful in preparing Triton X-100 acid urea (TAU) gel that has been developed for separating basic proteins such as histones and variants in acidic experimental condition. Additionally, the presence of titanium dioxide in the gel not only improves mechanistic property of gels but also changes the migration pattern of different proteins that have different affinities to titanium dioxide.

  4. Titanium Dioxide Photocatalytic Polymerization of Acrylamide for Gel Electrophoresis (TIPPAGE) of Proteins and Structural Identification by Mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Zhang, Wenyang; Yuan, Zhiwei; Huang, Lulu; Kang, Jie; Jiang, Ruowei; Zhong, Hongying

    2016-02-01

    Polyacrylamide gel electrophoresis (PAGE) coupled with mass spectrometry has been well established for separating, identifying and quantifying protein mixtures from cell lines, tissues or other biological samples. The copolymerization process of acrylamide and bis-acrylamide is the key to mastering this powerful technique. In general, this is a vinyl addition reaction initiated by free radical-generating reagents such as ammonium persulfate (APS) and tetramethylethylenediamine (TEMED) under basic pH and degassing experimental condition. We report herein a photocatalytic polymerization approach that is based on photo-generated hydroxyl radicals with nanoparticles of titanium dioxide. It was shown that the polymerization process is greatly accelerated in acidic condition when ultraviolet light shots on the gel solution containing TiO2 nanoparticles without degassing. This feature makes it very useful in preparing Triton X-100 acid urea (TAU) gel that has been developed for separating basic proteins such as histones and variants in acidic experimental condition. Additionally, the presence of titanium dioxide in the gel not only improves mechanistic property of gels but also changes the migration pattern of different proteins that have different affinities to titanium dioxide.

  5. Determination of DNA methylation by COBRA: a comparative study of CGE with LIF detection and conventional gel electrophoresis.

    PubMed

    Goedecke, Simon; Schlosser, Sabrina; Mühlisch, Jörg; Hempel, Georg; Frühwald, Michael C; Wünsch, Bernhard

    2009-09-01

    DNA methylation as an epigenetic modification of the human genome is under emphatic investigation. Several studies have demonstrated a role of DNA methylation in oncogenesis. In conjunction with histone modifications, DNA methylation may cause the formation of heterochromatin and thus mediate the inactivation of gene transcription. It is important to develop methods that allow for an accurate quantification of the amount of DNA methylation in particular DNA regions, to gain information concerning the threshold of methylation levels necessary for gene inactivation. In this article, a CGE method with on-column LIF detection using SYBR Green is compared with a conventional slab-gel electrophoresis. We thus investigate the validity to analyze DNA methylation in the samples of a combined bisulfite restriction analysis. It is demonstrated that CGE is superior to gel electrophoresis in means of linearity, precision, accuracy, automatization (high throughput), and sample consumption. However, gel electrophoresis is easier to perform (simple devices, no PC usage), and the running costs are comparatively low. A further advantage of CGE is the sparse use of toxic compounds (MeOH and SYBR Green), whereas gel electrophoresis is performed in polyacrylamide gels with ethidium bromide staining.

  6. Comparison of gel properties and biochemical characteristics of myofibrillar protein from bighead carp (Aristichthys nobilis) affected by frozen storage and a hydroxyl radical-generation oxidizing system.

    PubMed

    Lu, Han; Zhang, Longteng; Li, Qingzheng; Luo, Yongkang

    2017-05-15

    We wanted to clarify whether gel properties can be affected by in vivo or in vitro myofibrillar protein oxidation and, thus, to provide relevant information and a scientific foundation for the processing of gel products. To accomplish this, we measured the changes in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), total disulfide (SS) content, surface hydrophobicity (So-ANS), carbonyl content, and gel texture and water-holding capacity (WHC) of isolated myofibrillar protein from bighead carp fillets during frozen storage and under different H2O2 concentrations, which were used to represent in vivo and in vitro conditions, respectively. The results indicated that a certain range in content of disulfide crosslinks (0.91mol/10(5)g protein) would promote gel hardness. Mild protein oxidation caused by a certain degree of frozen storage and hydroxyl radicals can promote gel texture and WHC. Based on those results, freezing bighead carp for a certain period can be used to produce gel products.

  7. Sequence-specific nucleic acid mobility using a reversible block copolymer gel matrix and DNA amphiphiles (lipid-DNA) in capillary and microfluidic electrophoretic separations.

    PubMed

    Wagler, Patrick; Minero, Gabriel Antonio S; Tangen, Uwe; de Vries, Jan Willem; Prusty, Deepak; Kwak, Minseok; Herrmann, Andreas; McCaskill, John S

    2015-10-01

    Reversible noncovalent but sequence-dependent attachment of DNA to gels is shown to allow programmable mobility processing of DNA populations. The covalent attachment of DNA oligomers to polyacrylamide gels using acrydite-modified oligonucleotides has enabled sequence-specific mobility assays for DNA in gel electrophoresis: sequences binding to the immobilized DNA are delayed in their migration. Such a system has been used for example to construct complex DNA filters facilitating DNA computations. However, these gels are formed irreversibly and the choice of immobilized sequences is made once off during fabrication. In this work, we demonstrate the reversible self-assembly of gels combined with amphiphilic DNA molecules, which exhibit hydrophobic hydrocarbon chains attached to the nucleobase. This amphiphilic DNA, which we term lipid-DNA, is synthesized in advance and is blended into a block copolymer gel to induce sequence-dependent DNA retention during electrophoresis. Furthermore, we demonstrate and characterize the programmable mobility shift of matching DNA in such reversible gels both in thin films and microchannels using microelectrode arrays. Such sequence selective separation may be employed to select nucleic acid sequences of similar length from a mixture via local electronics, a basic functionality that can be employed in novel electronic chemical cell designs and other DNA information-processing systems.

  8. Dynamics of Swelling and Drying in a Spherical Gel

    NASA Astrophysics Data System (ADS)

    Bertrand, Thibault; Peixinho, Jorge; Mukhopadhyay, Shomeek; MacMinn, Christopher W.

    2016-12-01

    Swelling is a volumetric-growth process in which a porous material expands by spontaneous imbibition of additional pore fluid. Swelling is distinct from other growth processes in that it is inherently poromechanical: local expansion of the pore structure requires that additional fluid be drawn from elsewhere in the material, or into the material from across the boundaries. Here, we study the swelling and subsequent drying of a sphere of hydrogel. We develop a dynamic model based on large-deformation poromechanics and the theory of ideal elastomeric gels, and we compare the predictions of this model with a series of experiments performed with polyacrylamide spheres. We use the model and the experiments to study the complex internal dynamics of swelling and drying, and to highlight the fundamentally transient nature of these strikingly different processes. Although we assume spherical symmetry, the model also provides insight into the transient patterns that form and then vanish during swelling as well as the risk of fracture during drying.

  9. Detection and analysis of protein-protein interactions of organellar and prokaryotic proteomes by blue native and colorless native gel electrophoresis.

    PubMed

    Krause, Frank; Seelert, Holger

    2008-11-01

    Native gels enable the analysis of protein complexes on a proteome-wide scale in a single experiment. The protocols described in this unit are based on separation of protein complexes by blue native polyacrylamide electrophoresis (BN-PAGE), the most versatile native gel system, and the closely related milder colorless native PAGE (CN-PAGE). Both BN-PAGE and CN-PAGE are described on analytical to preparative scales. In addition, methods for subsequent analysis of protein complexes are given, including electroelution from native gels as well as denaturing and native two-dimensional PAGE. Finally, the removal of Coomassie dye from electroeluted proteins is detailed along with a discussion of fundamental considerations for the solubilization of membrane protein complexes from various biological samples, which are exemplified for mitochondria, chloroplasts (thylakoids), and cyanobacteria.

  10. Dynamics of a DNA Gel

    NASA Astrophysics Data System (ADS)

    Adhikari, Ramesh; Bhattacharya, Aniket; Dogariu, Aristide

    We study in silico the properties of a gel consisting of DNA strands (modeled as semi-flexible chains) and linkers of varying flexibility, length, and topology. These linkers are envisioned and modeled as active components with additional attributes so as to mimic properties of a synthetic DNA gel containing motor proteins. We use Brownian dynamics to directly obtain frequency dependent complex shear moduli of the gel. We further carry out force spectroscopy on these computer generated gels and study the relaxation properties as a function of the important parameters of the model, e.g., densities and relative ratios of the DNAs and the linkers, the average life time of a link, etc. Our studies are relevant for designing synthetic bio-materials for both materials and medical applications.

  11. Actuation performances of anisotropic gels

    NASA Astrophysics Data System (ADS)

    Nardinocchi, P.; Teresi, L.

    2016-12-01

    We investigated the actuation performances of anisotropic gels driven by mechanical and chemical stimuli, in terms of both deformation processes and stroke-curves, and distinguished between the fast response of gels before diffusion starts and the asymptotic response attained at the steady state. We also showed as the range of forces that an anisotropic hydrogel can exert when constrained is especially wide; indeed, changing fiber orientation allows us to induce shear as well as transversely isotropic extensions.

  12. Multiple phases of protien gels

    NASA Astrophysics Data System (ADS)

    Annaka, Masahiko; Tanaka, Toyoichi

    1994-03-01

    A multiple phase transition was observed in gels made by covalently cross-linking proteins in either native or denatured state. The enzymatic activity of the gels prepared from native α-chymotrypsin was determined for each of the multiple phases. The reversibility of the swelling degrees and the enzymatic reaction rates upon phase transition suggests that the protein is at a free energy minimum and thus in a phase.

  13. Topical Review: Polymer gel dosimetry

    PubMed Central

    Baldock, C; De Deene, Y; Doran, S; Ibbott, G; Jirasek, A; Lepage, M; McAuley, K B; Oldham, M; Schreiner, L J

    2010-01-01

    Polymer gel dosimeters are fabricated from radiation sensitive chemicals which, upon irradiation, polymerize as a function of the absorbed radiation dose. These gel dosimeters, with the capacity to uniquely record the radiation dose distribution in three-dimensions (3D), have specific advantages when compared to one-dimensional dosimeters, such as ion chambers, and two-dimensional dosimeters, such as film. These advantages are particularly significant in dosimetry situations where steep dose gradients exist such as in intensity-modulated radiation therapy (IMRT) and stereotactic radiosurgery. Polymer gel dosimeters also have specific advantages for brachytherapy dosimetry. Potential dosimetry applications include those for low-energy x-rays, high-linear energy transfer (LET) and proton therapy, radionuclide and boron capture neutron therapy dosimetries. These 3D dosimeters are radiologically soft-tissue equivalent with properties that may be modified depending on the application. The 3D radiation dose distribution in polymer gel dosimeters may be imaged using magnetic resonance imaging (MRI), optical-computerized tomography (optical-CT), x-ray CT or ultrasound. The fundamental science underpinning polymer gel dosimetry is reviewed along with the various evaluation techniques. Clinical dosimetry applications of polymer gel dosimetry are also presented. PMID:20150687

  14. TOPICAL REVIEW: Polymer gel dosimetry

    NASA Astrophysics Data System (ADS)

    Baldock, C.; De Deene, Y.; Doran, S.; Ibbott, G.; Jirasek, A.; Lepage, M.; McAuley, K. B.; Oldham, M.; Schreiner, L. J.

    2010-03-01

    Polymer gel dosimeters are fabricated from radiation sensitive chemicals which, upon irradiation, polymerize as a function of the absorbed radiation dose. These gel dosimeters, with the capacity to uniquely record the radiation dose distribution in three-dimensions (3D), have specific advantages when compared to one-dimensional dosimeters, such as ion chambers, and two-dimensional dosimeters, such as film. These advantages are particularly significant in dosimetry situations where steep dose gradients exist such as in intensity-modulated radiation therapy (IMRT) and stereotactic radiosurgery. Polymer gel dosimeters also have specific advantages for brachytherapy dosimetry. Potential dosimetry applications include those for low-energy x-rays, high-linear energy transfer (LET) and proton therapy, radionuclide and boron capture neutron therapy dosimetries. These 3D dosimeters are radiologically soft-tissue equivalent with properties that may be modified depending on the application. The 3D radiation dose distribution in polymer gel dosimeters may be imaged using magnetic resonance imaging (MRI), optical-computerized tomography (optical-CT), x-ray CT or ultrasound. The fundamental science underpinning polymer gel dosimetry is reviewed along with the various evaluation techniques. Clinical dosimetry applications of polymer gel dosimetry are also presented.

  15. Rheological characterization of hydroxypropylcellulose gels.

    PubMed

    Ramachandran, S; Chen, S; Etzler, F

    1999-02-01

    The present paper describes the rheological properties of hydroxypropylcellulose (HPC) gels formulated in propylene glycol (PG), water, ethanol, and mixtures of these components. The effects of molecular weight, polymer concentration, and solvent composition on the apparent viscosity and flow characteristics have been studied by continuous shear rheometry. The HPC gels are shear thinning and do not exhibit significant yield or hysteresis in their rheograms. The apparent viscosity increases with increasing molecular weight and concentration of the polymer, as expected. Although not so pronounced at lower concentrations (< or = 1.5%), HPC gels tend to become increasingly non-Newtonian with increasing molecular weight at higher polymer concentrations (3%). A mathematical model has been proposed for the prediction of viscosities of HPC gels. There exists a high degree of dependence on molecular interactions between various solvent molecules in the prediction of mixture viscosities in ternary systems. The effects of solvent composition on the viscoelastic behavior of these gels have also been examined by dynamic mechanical analysis. The HPC gels are highly viscoelastic and exhibit greater degrees of elasticity with increased PG content in ternary solvent mixtures with water and ethanol. The study also suggests that dynamic mechanical analysis could prove to be a useful tool in the determination of zero-shear viscosities, viscosities that are representative of most realistic situations.

  16. Application of chitosan/polyacrylamide nanofibres for removal of chromate and phosphate in water

    NASA Astrophysics Data System (ADS)

    Nthumbi, Richard M.; Catherine Ngila, J.; Moodley, Brenda; Kindness, Andrew; Petrik, Leslie

    Water pollution is an intractable environmental problem in South Africa. Management of the water resource is vital in order to address the water scarcity issues. Research on remediation of contaminated water has focused mainly on the removal of heavy metals such as Pb, Cd, Zn, Hg and Cu and neglected other inorganic pollutants. In this work we focus on the removal of anions, namely chromate and phosphate. Chromium is extensively used in the textile, leather and metallurgy industries and contaminates surface water and groundwater when inadequately treated industrial effluents are discharged. Chromium has been associated with irregular sugar metabolism, nosebleeds and ulcers, and it is also carcinogenic. The phosphate ion is an essential micronutrient responsible for healthy plant growth. However, excess phosphate intake stimulates rapid growth of photosynthetic algae and cyanobacteria, resulting in eutrophication. This phenomenon (algal bloom) causes other organisms to die due to reduced oxygen in the water. In order to offer remediation measures, this study reports the use of electrospun nanofibres for the removal of chromate and phosphate anions. Adsorption experiments were carried out using nanofibres electrospun from chitosan and polyacrylamide polymer blends, cross-linked with glutaraldehyde. Quantification of chromium was done using ICP-OES while UV-Vis spectrophotometry was used for the determination of phosphates. Batch adsorption experiments were done to determine optimum adsorption parameters such as pH, contact time, temperature and initial analyte concentration. Removal of the ions using a flow-adsorption technique through a micro-column was performed. The experimental data obtained were analysed using Langmuir and Freundlich models to study the adsorption mechanisms. The nanofibres had an adsorption capacity for Cr(VI) and PO43- of 0.26 mg g-1 and 392 mg g-1, respectively, and removal efficiencies of 93% and 97.4%, in the same order, in synthetic water

  17. Polyacrylamide hydrogels: an effective tool for delivering liquid baits to pest ants (Hymenoptera: Formicidae).

    PubMed

    Buczkowski, Grzegorz; Roper, Elray; Chin, Darren

    2014-04-01

    Ant management in urban and natural areas often relies on toxic baits. Liquid baits are highly attractive to pest ants because they mimic natural food sources such as honeydew and nectar, the principal dietary components of many ants. However, liquid bait use has been limited owing to the lack of bait dispensers that are effective, inexpensive, and easy to service. The current study evaluated the potential of water-storing crystals (polyacrylamide spheres) to effectively deliver liquid thiamethoxam baits to laboratory colonies of Argentine ants, Linepithema humile Mayr. Results of laboratory trials show that bait crystals saturated in 25% sucrose solution containing 0.007% thiamethoxam are highly attractive to Argentine ants and highly effective against all castes and life stages, including workers, queens, and brood. Fresh bait crystals were highly effective and required approximately 2 d to kill all workers and approximately 6 d to achieve complete mortality in queens and brood. Results of bait aging tests show that the crystals lose approximately 70% of moisture in 8 h and the duration of outdoor exposure has a significant effect on moisture loss and subsequently bait acceptance and bait efficacy. A gradual decrease in mortality was observed for all castes and life stages as bait age increased. In general, fresh baits and those aged for < 8 h retained their efficacy and caused substantial mortality. Baits aged longer than 8 h were substantially less attractive and less effective. Horizontal transfer tests examined the transfer of thiamethoxam from live treated donors to live untreated recipients. The results show that donor ants that obtain thiamethoxam by feeding on bait crystals effectively transfer it to untreated recipient ants. The level of secondary mortality depended on the donor:recipient ratio, with approximately 40% recipient worker mortality with the 1:5 ratio and 15% recipient worker mortality with 1:10 or 1:20 ratios. However, no queens died in any

  18. 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.

  19. Electrophoresis for genotyping: temporal thermal gradient gel electrophoresis for profiling of oligonucleotide dissociation.

    PubMed Central

    Day, I N; O'Dell, S D; Cash, I D; Humphries, S E; Weavind, G P

    1995-01-01

    Traditional use of an oligonucleotide probe to determine genotype depends on perfect base pairing to a single-stranded target which is stable to a higher temperature than when imperfect binding occurs due to a mismatch in the target sequence. Bound oligonucleotide is detected at a predetermined single temperature 'snapshot' of the melting profile, allowing the distinction of perfect from imperfect base pairing. In heterozygotes, the presence of the alternative sequence must be verified with a second oligonucleotide complementary to the variant. Here we describe a system of real-time variable temperature electrophoresis during which the oligonucleotide dissociates from its target. In 20% polyacrylamide the target strand has minimal mobility and released oligonucleotide migrates extremely quickly so that the 'freed' rather than the 'bound' is displayed. The full profile of oligonucleotide dissociation during gel electrophoresis is represented along the gel track, and a single oligonucleotide is sufficient to confirm heterozygosity, since the profile displays two separate peaks. Resolution is great, with use of short track lengths enabling analysis of dense arrays of samples. Each gel track can contain a different target or oligonucleotide and the temperature gradient can accommodate oligonucleotides of different melting temperatures. This provides a convenient system to examine the interaction of many different oligonucleotides and target sequences simultaneously and requires no prior knowledge of the mutant sequence(s) nor of oligonucleotide melting temperatures. The application of the technique is described for screening of a hotspot for mutations in the LDL receptor gene in patients with familial hypercholesterolaemia. Images PMID:7630718

  20. Single Element 2-DIMENSIONAL Acousto-Optic Deflectors Design, Fabrication and Implementation for Digital Optical Computing

    NASA Astrophysics Data System (ADS)

    Rosemeier, Jolanta Iwona

    1992-09-01

    With the need to develop very fast computers compared to the conventional digital chip based systems, the future is very bright for optical based signal processing. Attention has turned to a different application of optics utilizing mathematical operations, in which case operations are numerical, sometimes discrete, and often algebraic in nature. Interest has been so vigorous that many view it as a small revolution in optics whereby optical signal processing is beginning to encompass what many frequently describe as optical computing. The term is fully intended to imply close comparison with the operations performed by scientific digital computers. Most present computer intensive problem solving processors rely on a common set of linear equations found in numerical matrix algebra. Recently, considerable research focused on the use of systolic array, which can operate at high speeds with great efficiency. This approach addresses the acousto-optic digital and analog arrays utilizing three dimensional optical interconnect technology. In part I of this dissertation the first single element 2-dimensional (2-D) acousto-optic deflector was designed, fabricated and incorporated into an optical 3 x 3 vector-vector or matrix-matrix multiplier system. This single element deflector is used as a outer-product device. The input vectors are addressed by electronic means and the outer product matrix is displayed as a 2-D array of optical (laser) pixels. In part II of this work a multichannel single element 2-D deflector was designed, fabricated and implemented into a Programmable Logic Array (PLA) optical computing system. This system can be used for: word equality detection, free space optical interconnections, half adder optical system implementation. The PLA system described in this dissertation has capability of word equality detection. The 2-D multichannel deflector that was designed and fabricated is capable of comparing 16 x 16 words every 316 nanoseconds. Each word is 8

  1. Bleach gel: a simple agarose gel for analyzing RNA quality.

    PubMed

    Aranda, Patrick S; LaJoie, Dollie M; Jorcyk, Cheryl L

    2012-01-01

    RNA-based applications requiring high-quality, non-degraded RNA are a foundational element of many research studies. As such, it is paramount that the integrity of experimental RNA is validated prior to cDNA synthesis or other downstream applications. In the absence of expensive equipment such as microfluidic electrophoretic devices, and as an alternative to the costly and time-consuming standard formaldehyde gel, RNA quality can be quickly analyzed by adding small amounts of commercial bleach to TAE buffer-based agarose gels prior to electrophoresis. In the presence of low concentrations of bleach, the secondary structure of RNA is denatured and potential contaminating RNases are destroyed. Because of this, the 'bleach gel' is a functional approach that addresses the need for an inexpensive and safe way to evaluate RNA integrity and will improve the ability of researchers to rapidly analyze RNA quality.

  2. Thixotropic gel for vadose zone remediation

    DOEpatents

    Riha, Brian D.

    2012-07-03

    A thixotropic gel suitable for use in subsurface bioremediation is provided along with a process of using the gel. The thixotropic gel provides a non-migrating injectable substrate that can provide below ground barrier properties. In addition, the gel components provide for a favorable environment in which certain contaminants are preferentially sequestered in the gel and subsequently remediated by either indigenous or introduced microorganisms.

  3. Thixotropic gel for vadose zone remediation

    DOEpatents

    Rhia, Brian D [Augusta, GA

    2011-03-01

    A thixotropic gel suitable for use in subsurface bioremediation is provided along with a process of using the gel. The thixotropic gel provides a non-migrating injectable substrate that can provide below ground barrier properties. In addition, the gel components provide for a favorable environment in which certain contaminants are preferentially sequestered in the gel and subsequently remediated by either indigenous or introduced microorganisms.

  4. Thixotropic gel for vadose zone remediation

    SciTech Connect

    Riha, Brian D.; Looney, Brian B.

    2015-10-27

    A thixotropic gel suitable for use in subsurface bioremediation is provided along with a process of using the gel. The thixotropic gel provides a non-migrating injectable substrate that can provide below ground barrier properties. In addition, the gel components provide for a favorable environment in which certain contaminants are preferentially sequestered in the gel and subsequently remediated by either indigenous or introduced microorganisms.

  5. Microwave assisted synthesis of polyacrylamide grafted starch (St-g-PAM) and its applicability as flocculant for water treatment.

    PubMed

    Mishra, Sumit; Mukul, Ankita; Sen, Gautam; Jha, Usha

    2011-01-01

    Polyacrylamide grafted starch (St-g-PAM) was made by a novel method of synthesis, involving combination of microwave radiation and a chemical free radical initiator (ceric ammonium nitrate) to initiate grafting reaction. This method (microwave assisted synthesis) is quick, highly reliable, reproducible and yields high quality product as compared to the conventional method (which uses a chemical free radical initiator alone to initiate the grafting reaction).The St-g-PAM grades synthesized were characterized by various physicochemical techniques. Further, its application as flocculant for wastewater treatment was investigated.

  6. Deformation of Unentangled Swollen Gels

    NASA Astrophysics Data System (ADS)

    Sariyer, Ozan; Panyukov, Sergey; Rubinstein, Michael

    2014-03-01

    We study the deformation characteristics (Poisson's ratios and stress-strain relations) of unentangled gels swollen and uniaxially or biaxially deformed in excess solvent by considering the balance of osmotic pressure and elastic stress in unconstrained dimensions. Our scaling theory predicts a crossover from theta solvent behavior to marginal solvent behavior upon stretching gels that are in concentrated regime at swelling equilibrium - a phenomenon that was experimentally observed long ago, but not understood theoretically. For gels that are in the semidilute good solvent regime at swelling equilibrium, we predict a crossover to theta solvent behavior upon compression and a crossover to marginal solvent behavior upon stretching. Our theory reproduces the previously known results for equilibrium swelling degree as well as known deformation characteristics in theta and athermal solvents.

  7. Sol-gel derived sorbents

    DOEpatents

    Sigman, Michael E.; Dindal, Amy B.

    2003-11-11

    Described is a method for producing copolymerized sol-gel derived sorbent particles for the production of copolymerized sol-gel derived sorbent material. The method for producing copolymerized sol-gel derived sorbent particles comprises adding a basic solution to an aqueous metal alkoxide mixture for a pH.ltoreq.8 to hydrolyze the metal alkoxides. Then, allowing the mixture to react at room temperature for a precalculated period of time for the mixture to undergo an increased in viscosity to obtain a desired pore size and surface area. The copolymerized mixture is then added to an immiscible, nonpolar solvent that has been heated to a sufficient temperature wherein the copolymerized mixture forms a solid upon the addition. The solid is recovered from the mixture, and is ready for use in an active sampling trap or activated for use in a passive sampling trap.

  8. Copolymers For Capillary Gel Electrophoresis

    SciTech Connect

    Liu, Changsheng; Li, Qingbo

    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.

  9. An investigation of cutting mechanics in 2 dimensional ultrasonic vibration assisted milling toward chip thickness and chip formation

    NASA Astrophysics Data System (ADS)

    Rasidi, I. I.; Rafai, N. H.; Rahim, E. A.; Kamaruddin, S. A.; Ding, H.; Cheng, K.

    2015-12-01

    The purpose of this paper is to investigate the effects of 2 dimensional Ultrasonic Vibration Assisted Milling (UVAM) cutting mechanics, considering tool path trajectory and the effect on the chip thickness. The theoretical modelling of cutting mechanics is focused by considering the trajectory of the tool locus into the workpiece during the machining. The studies found the major advantages of VAM are come from the intermittent tool tip interaction phenomena between cutting tool and workpiece. The reduction of thinning chip thickness formations can be identifying advantages from vibration assisted milling in 2 dimensional. The finding will be discussing the comparison between conventional machining the potential of the advantages toward the chip thickness and chip formation in conclusion.

  10. Supramolecular Construction of Multifluorescent Gels: Interfacial Assembly of Discrete Fluorescent Gels through Multiple Hydrogen Bonding.

    PubMed

    Ji, Xiaofan; Shi, Bingbing; Wang, Hu; Xia, Danyu; Jie, Kecheng; Wu, Zi Liang; Huang, Feihe

    2015-12-22

    Multifluorescent supramolecular gels with complex structures are constructed from discrete fluorescent gels, which serve as the building blocks, through hydrogen bonding interactions at interfaces. The multifluorescent gel can realize rapid healing within only ≈100 s.

  11. Microbubble tunneling in gel phantoms

    PubMed Central

    Caskey, Charles F.; Qin, Shengping; Dayton, Paul A.; Ferrara, Katherine W.

    2009-01-01

    Insonified microbubbles were observed in vessels within a gel with a Young’s modulus similar to that of tissue, demonstrating shape instabilities, liquid jets, and the formation of small tunnels. In this study, tunnel formulation occurred in the direction of the propagating ultrasound wave, where radiation pressure directed the contact of the bubble and gel, facilitating the activity of the liquid jets. Combinations of ultrasonic parameters and microbubble concentrations that are relevant for diagnostic imaging and drug delivery and that lead to tunnel formation were applied and the resulting tunnel formation was quantified. PMID:19425620

  12. Statistical physics of polymer gels

    NASA Astrophysics Data System (ADS)

    Panyukov, Sergei; Rabin, Yitzhak

    1996-05-01

    This work presents a comprehensive analysis of the statistical mechanics of randomly cross-linked polymer gels, starting from a microscopic model of a network made of instantaneously cross-linked Gaussian chains with excluded volume, and ending with the derivation of explicit expressions for the thermodynamic functions and for the density correlation functions which can be tested by experiments. Using replica field theory we calculate the mean field density in replica space and show that this solution contains statistical information about the behavior of individual chains in the network. The average monomer positions change affinely with macroscopic deformation and fluctuations about these positions are limited to length scales of the order of the mesh size. We prove that a given gel has a unique state of microscopic equilibrium which depends on the temperature, the solvent, the average monomer density and the imposed deformation. This state is characterized by the set of the average positions of all the monomers or, equivalently, by a unique inhomogeneous monomer density profile. Gels are thus the only known example of equilibrium solids with no long-range order. We calculate the RPA density correlation functions that describe the statistical properties of small deviations from the average density, due to both static spatial heterogeneities (which characterize the inhomogeneous equilibrium state) and thermal fluctuations (about this equilibrium). We explain how the deformation-induced anisotropy of the inhomogeneous equilibrium density profile is revealed by small angle neutron scattering and light scattering experiments, through the observation of the butterfly effect. We show that all the statistical information about the structure of polymer networks is contained in two parameters whose values are determined by the conditions of synthesis: the density of cross-links and the heterogeneity parameter. We find that the structure of instantaneously cross

  13. Improving Water Quality in Construction Site Runoff: Optimal Mixing Time and Dose for Flocculating Suspended Sediment with Polyacrylamide

    NASA Astrophysics Data System (ADS)

    Zare, A. H.

    2015-12-01

    Sediment, a major water pollutant, can harm ecosystems and water resources. Sediment in construction site runoff can be controlled through flocculation using anionic, linear polyacrylamide (PAM), but there is little information on optimizing these applications. We conducted laboratory experiments to determine the optimal mixing times for varying concentrations of two types of polyacrylamide, APS 705 and FA 920, which are blends of polymers with a range of molecular weights that are anionic and neutral, respectively. These were selected from a variety of PAMs previously screened for flocculation potential. Soil from three active construction sites in the Piedmont region of North Carolina were used in the testing. A mixing speed of 300 revolutions per minute (RPM), the maximum available for the paddle mixer we used, was the most effective at reducing turbidity with PAM. Turbidity was reduced with increased mixing times up to a point, after which little additional benefit was evident. For all three soils tested, turbidity decreased as mixing time reached 1-2 minutes at polymer doses of 1, 5 and 10 mg L-1, with no substantial reduction with further mixing. At a polymer dose of 0.5 mg L-1, however, turbidity tended to increase beyond 5 minutes of mixing time, possibly because excessive shear forces destroyed sparsely linked floc. For both PAMs, 1 mg L-1 and a mixing time of 2-3 min appeared to be sufficient to achieve the most effective turbidity reduction.

  14. Removal and separation of Cu(II) from aqueous solutions using nano-silver chitosan/polyacrylamide membranes.

    PubMed

    Abou El-Reash, Y G; Abdelghany, A M; Elrazak, A Abd

    2016-05-01

    In the present study, adsorption of Cu(II) ions from aqueous solutions was evaluated using new thin adsorptive membranes modified with silver nanoparticles. Membranes were prepared from chitosan/polyacrylamide polymer blend. The variation of adsorption process was investigated in batch sorption mode. Infrared absorption spectra were applied for chemical characterization of the prepared polymer blend. Thermogravimetric analysis showed that addition of polyacrylamide to chitosan increased its thermal stability. The kinetics and thermodynamic parameters of Cu(II) ions adsorption onto the membranes were studied by removal experiments of Cu(II) ions from standard aqueous solutions. The kinetic data fitted to the traditional Lagergren adsorption kinetic equations. Thermodynamic studies indicated endothermic (ΔH°> 0) and spontaneous (ΔG°< 0) adsorption together with entropy generation (ΔS°> 0) at the solid/liquid interface process. Regeneration experiments showed that the newly prepared membranes could be reconditioned without significant loss of its initial properties even after three adsorption-desorption cycles. The results suggest that the prepared composite membranes can be efficiently applied for the adsorptive removal of Cu(II) ions from natural water samples. Antimicrobial activity was tested against two gram negatives, two gram positives and Candida sp. microbes and they showed a remarkable bioactivity indicating the capability of applying such membranes for a dual action.

  15. Sol-gel transition of nanoparticles/polymer mixtures for sustained delivery of exenatide to treat type 2 diabetes mellitus.

    PubMed

    Oh, Keun Sang; Kim, Jae Yeon; Yoon, Byeong Deok; Lee, Minae; Kim, Heejoo; Kim, Michelle; Seo, Jae Hong; Yuk, Soon Hong

    2014-11-01

    The sol-gel transition of nanoparticles (NPs)/polymer mixture in aqueous medium was investigated for the sustained delivery of exenatide to treat type 2 diabetes mellitus. Exenatide-loaded multilayer NPs were prepared using a layer-by-layer approach which utilized the interaction between Pluronics and lipid bilayers as the main driving force for the construction of the multilayer. Pluronic F-127 was the polymer used, and it forms a gel at body temperature. Although the antidiabetic effects of exenatide-loaded multilayer NPs have been demonstrated previously in an animal model, in this work, the attempt was made to demonstrate the extended duration of antidiabetic effects, which was accomplished by localizing the exenatide-loaded NPs in muscular areas in the body through the gelation of Pluronic F-127. Transmittance electron microscopy and dynamic light scattering were used to examine the morphology of the multilayer NPs/polymer mixture. A change in the release pattern of exenatide was observed after gel formation at body temperature, and Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis was performed using native exenatide and a reference biomarker as control to observe whether exenatide extracted from the multilayer NPs and the multilayer NPs/Pluronic F-127 mixture degraded or not. We then observed the antidiabetic effect of exenatide-loaded multilayer NPs/Pluronic F-127 mixture by monitoring blood-glucose levels in db/db mice. In vitro and in vivo correlation was discussed regarding structural variation in the delivery vehicles.

  16. AraUTP-Affi-Gel 10: a novel affinity absorbent for the specific purification of DNA polymerase alpha-primase.

    PubMed

    Izuta, S; Saneyoshi, M

    1988-10-01

    For the specific purification of eukaryotic DNA-dependent DNA polymerase alpha, we prepared two novel affinity resins bearing 5-(E)-(4-aminostyryl) araUTP as a ligand. One of them was araUTP-Sepharose 4B which was coupled directly with the ligand and the other was araUTP-Affi-Gel 10 which was coupled with the ligand through a spacer. No DNA polymerase alpha-primase activity from cherry salmon (Oncorhynchus masou) testes was bound on the araUTP-Sepharose 4B in all cases examined. On the other hand, the araUTP-Affi-Gel 10 retains this enzyme activity when poly(dA) or poly(dA)-oligo(dT)12-18 is present. The retained enzyme activity was sharply eluted around 100-mM KCl concentrations as a single peak, and this fraction showed a specific activity of about 170,000 units/mg as alpha-polymerase activity. The highly purified DNA polymerase alpha-primase isolated using the araUTP-Affi-Gel 10 contained only three polypeptides, which showed Mr values of 120,000, 62,000, and 58,000, respectively, as judged using sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

  17. Nonlinear elasticity of alginate gels

    NASA Astrophysics Data System (ADS)

    Hashemnejad, Seyed Meysam; Kundu, Santanu

    Alginate is a naturally occurring anionic polysaccharide extracted from brown algae. Because of biocompatibility, low toxicity, and simple gelation process, alginate gels are used in biomedical and food applications. Here, we report the rheological behavior of ionically crosslinked alginate gels, which are obtained by in situ gelation of alginates with calcium salts, in between two parallel plates of a rheometer. Strain stiffening behavior was captured using large amplitude oscillatory shear (LAOS) experiments. In addition, negative normal stress was observed for these gels, which has not been reported earlier for any polysaccharide networks. The magnitude of negative normal stress increases with applied strain and can exceed that of the shear stress at large strain. Rheological results fitted with a constitutive model that considers both stretching and bending of chains indicate that nonlinearity is likely related to the stretching of the chains between the crosslink junctions. The results provide an improved understanding of the deformation mechanism of ionically crosslinked alginate gel and the results will be important in developing synthetic extracellular matrix (ECM) from these materials.

  18. Liquid-crystalline physical gels.

    PubMed

    Kato, Takashi; Hirai, Yuki; Nakaso, Suguru; Moriyama, Masaya

    2007-12-01

    Liquid-crystalline (LC) physical gels are a new class of dynamically functional materials consisting of liquid crystals and fibrous aggregates of molecules that are called "gelators". Liquid-crystalline physical gels, which are macroscopically soft solids, exhibit induced or enhanced electro-optical, photochemical, electronic properties due to the combination of two components that form phase-separated structures. In this tutorial review, we describe the materials design and structure-property relationships of the LC physical gels. The introduction of self-assembled fibers into nematic liquid crystals leads to faster responses in twisted nematic (TN) mode and high contrast switching in light scattering mode. Furthermore, the LC physical gels can be exploited as a new type of materials for electro-optical memory. This function is achieved by the control of reversible aggregation processes of gelators under electric fields in nematic liquid crystals. Electronic properties such as hole mobilities are improved by the introduction of fibrous aggregates into triphenylene-based columnar liquid crystals. The incorporation of photochromic azobenzenes or electroactive tetrathiafulvalenes into the chemical structures of gelators leads to the preparation of ordered functional materials.

  19. New frontiers in materials science for art conservation: responsive gels and beyond.

    PubMed

    Carretti, Emiliano; Bonini, Massimo; Dei, Luigi; Berrie, Barbara H; Angelova, Lora V; Baglioni, Piero; Weiss, Richard G

    2010-06-15

    -flowing liquid can be removed gently. (2) Incorporation of magnetic, coated-ferrite nanoparticles into polyacrylamide gels adds functionality to a versatile system comprising oil-in-water microemulsions, aqueous micellar solutions, or xerogels that act as sponges. The ferrite particles allow the use of magnets both to place the gels precisely on a surface and to lift them from it after cleaning. (3) Novel formulations of poly(vinyl alcohol)-borate gels, which accept a range of organic cosolvents, show promise for swelling and dissolving organic coatings. This family of gels can be quite stiff but can be spread. They are non-sticky and have sufficient strength to be removed by peeling or lifting them from a sensitive surface. These three classes of gels are potentially very important soft materials to augment and improve the range of options available for conserving cultural heritage, and their interesting chemical-physical properties open a rich area for future scientific investigation.

  20. Enhanced and selective adsorption of mercury ions on chitosan beads grafted with polyacrylamide via surface-initiated atom transfer radical polymerization.

    PubMed

    Li, Nan; Bai, Renbi; Liu, Changkun

    2005-12-06

    Enhanced and selective removal of mercury ions was achieved with chitosan beads grafted with polyacrylamide (chitosan-g-polyacrylamide) via surface-initiated atom transfer radical polymerization (ATRP). The chitosan-g-polyacrylamide beads were found to have significantly greater adsorption capacities and faster adsorption kinetics for mercury ions than the chitosan beads. At pH 4 and with initial mercury concentrations of 10-200 mg/L, the chitosan-g-polyacrylamide beads can achieve a maximum adsorption capacity of up to 322.6 mg/g (in comparison with 181.8 mg/g for the chitosan beads) and displayed a short adsorption equilibrium time of less than 60 min (compared to more than 15 h for the chitosan beads). Coadsorption experiments with both mercury and lead ions showed that the chitosan-g-polyacrylamide beads had excellent selectivity in the adsorption of mercury ions over lead ions at pH < 6, in contrast to the chitosan beads, which did not show clear selectivity for either of the two metal species. Mechanism study suggested that the enhanced mercury adsorption was due to the many amide groups grafted onto the surfaces of the beads, and the selectivity in mercury adsorption can be attributed to the ability of mercury ions to form covalent bonds with the amide. It was found that adsorbed mercury ions on the chitosan-g-polyacrylamide beads can be effectively desorbed in a perchloric acid solution, and the regenerated beads can be reused almost without any loss of adsorption capacity.

  1. Bleach Gel: A Simple Agarose Gel for Analyzing RNA Quality

    PubMed Central

    Aranda, Patrick S.; LaJoie, Dollie M.; Jorcyk, Cheryl L.

    2013-01-01

    RNA-based applications requiring high quality, non-degraded RNA are a foundational element of many research studies. As such, it is paramount that the integrity of experimental RNA is validated prior to cDNA synthesis or other downstream applications. In the absence of expensive equipment such as microfluidic electrophoretic devices, and as an alternative to the costly and time-consuming standard formaldehyde gel, RNA quality can be quickly analyzed by adding small amounts of commercial bleach to TAE buffer-based agarose gels prior to electrophoresis. In the presence of low concentrations of bleach, the secondary structure of RNA is denatured and potential contaminating RNases are destroyed. Because of this, the ‘bleach gel’ is a functional approach that addresses the need for an inexpensive and safe way to evaluate RNA integrity and will improve the ability of researchers to rapidly analyze RNA quality. PMID:22222980

  2. Clindamycin phosphate 1% gel in acne vulgaris.

    PubMed

    Rizer, R L; Sklar, J L; Whiting, D; Bucko, A; Shavin, J; Jarratt, M

    2001-01-01

    A 12-week study compared Clindagel, a unique water-based gel formulation of clindamycin phosphate 1%, administered once daily, and Cleocin T, a slightly different gel formulation indicated for twice-daily use, in the treatment of acne vulgaris. Clindagel was safe and effective and equivalent to Cleocin T gel, albeit with a better tolerability profile. Clindagel is a viable alternative to Cleocin T gel.

  3. Study of Fricke gel dosimeter response for different gel quality

    NASA Astrophysics Data System (ADS)

    Cavinato, C. C.; Campos, L. L.

    2010-11-01

    The Fricke xylenol gel (FXG) dosimeter has been studied for application in radiotherapy because it is capable of to measure the spatial distribution of radiation doses. The dosimetry is based on the oxidation of ferrous (Fe2+) to ferric (Fe3+) ions radiation induced, related to the radiation dose. The gel material usually employed is the 300 Bloom gelatin, which is imported and very expensive in Brazil. Aiming to analyze the viability of to use a locally produced and low cost gel material, in this work the spectrophotometric responses of FXG solutions prepared using 270 Bloom gelatin commercially available and 300 Bloom gelatin imported were compared. The absorption spectra of solutions prepared with 5% by weight 270 and 300 Bloom gelatins non-irradiated and irradiated with 60Co gamma radiation in the dose range between 0.5 and 100 Gy were analysed, the dose-response curves were evaluated and the useful dose range was established. The obtained results indicate that the FXG solution prepared with 270 Bloom gelatin presents good performance, similar to that presented by the FXG solution prepared with 300 Bloom gelatin and its use can be recommended owing to the low cost and the availability in local market.

  4. DNA electrophoresis in agarose gels: Effects of electric field and gel concentration on the exponential dependence of reciprocal mobility on DNA length

    NASA Astrophysics Data System (ADS)

    Beheshti, Afshin; van Winkle, David; Randolph, Rill

    2002-03-01

    Electrophoresis was performed on double stranded DNA fragments ranging in length from 200 bp to 48502 bp at agarose gel concentrations T = 0.5% - 1.5% and electric fields E = 0.71 V/cm to 5 V/cm. A wide range of electric fields and gel concentrations were used to find what range of conditions work with the new interpolation equation, 1/μ(L) = 1/μl - (1/μl - 1/μ_s)e^-L/γ. The equation fit extremely well (\\chi^2 >= 0.999) to data with E = 2.5 V/cm to 5 V/cm and for lower fields (E < 2.5 V/cm) at low gel concentrations (T = 0.5% and 0.7%). This exponential relation seemed to hold when there is a smooth transition from the Ogston sieving regime to the reptation regime when looking at the “reptation plots” (plotting 3μL/μo vs. L) (Rousseau, J., Drouin, G., and Slater, G. W., Phys Rev Lett. 1997, 79, 1945-1948). For separations of single-stranded DNA in polyacrylamide, similar reptation plots have a region with a negative slope between the Ogston sieving regime and the reptation regime which has been interpreted as the signature of entropic trapping. When separating double-stranded DNA in agarose it was observed that fits deviate from the data when three different slopes are observed in the reptation plots. Failure of the simple exponential relationship between reciprocal mobility and DNA length appears to be the consequence of entropic trapping.

  5. Classification of 2-dimensional array patterns: assembling many small neural networks is better than using a large one.

    PubMed

    Chen, Liang; Xue, Wei; Tokuda, Naoyuki

    2010-08-01

    In many pattern classification/recognition applications of artificial neural networks, an object to be classified is represented by a fixed sized 2-dimensional array of uniform type, which corresponds to the cells of a 2-dimensional grid of the same size. A general neural network structure, called an undistricted neural network, which takes all the elements in the array as inputs could be used for problems such as these. However, a districted neural network can be used to reduce the training complexity. A districted neural network usually consists of two levels of sub-neural networks. Each of the lower level neural networks, called a regional sub-neural network, takes the elements in a region of the array as its inputs and is expected to output a temporary class label, called an individual opinion, based on the partial information of the entire array. The higher level neural network, called an assembling sub-neural network, uses the outputs (opinions) of regional sub-neural networks as inputs, and by consensus derives the label decision for the object. Each of the sub-neural networks can be trained separately and thus the training is less expensive. The regional sub-neural networks can be trained and performed in parallel and independently, therefore a high speed can be achieved. We prove theoretically in this paper, using a simple model, that a districted neural network is actually more stable than an undistricted neural network in noisy environments. We conjecture that the result is valid for all neural networks. This theory is verified by experiments involving gender classification and human face recognition. We conclude that a districted neural network is highly recommended for neural network applications in recognition or classification of 2-dimensional array patterns in highly noisy environments.

  6. Analysis of Dictyostelium discoideum inositol pyrophosphate metabolism by gel electrophoresis.

    PubMed

    Pisani, Francesca; Livermore, Thomas; Rose, Giuseppina; Chubb, Jonathan Robert; Gaspari, Marco; Saiardi, Adolfo

    2014-01-01

    The social amoeba Dictyostelium discoideum was instrumental in the discovery and early characterization of inositol pyrophosphates, a class of molecules possessing highly-energetic pyrophosphate bonds. Inositol pyrophosphates regulate diverse biological processes and are attracting attention due to their ability to control energy metabolism and insulin signalling. However, inositol pyrophosphate research has been hampered by the lack of simple experimental procedures to study them. The recent development of polyacrylamide gel electrophoresis (PAGE) and simple staining to resolve and detect inositol pyrophosphate species has opened new investigative possibilities. This technology is now commonly applied to study in vitro enzymatic reactions. Here we employ PAGE technology to characterize the D. discoideum inositol pyrophosphate metabolism. Surprisingly, only three major bands are detectable after resolving acidic extract on PAGE. We have demonstrated that these three bands correspond to inositol hexakisphosphate (IP₆ or Phytic acid) and its derivative inositol pyrophosphates, IP₇ and IP₈. Biochemical analyses and genetic evidence were used to establish the genuine inositol phosphate nature of these bands. We also identified IP₉ in D. discoideum cells, a molecule so far detected only from in vitro biochemical reactions. Furthermore, we discovered that this amoeba possesses three different inositol pentakisphosphates (IP₅) isomers, which are largely metabolised to inositol pyrophosphates. Comparison of PAGE with traditional Sax-HPLC revealed an underestimation of the cellular abundance of inositol pyrophosphates by traditional methods. In fact our study revealed much higher levels of inositol pyrophosphates in D. discoideum in the vegetative state than previously detected. A three-fold increase in IP₈ was observed during development of D. discoideum a value lower that previously reported. Analysis of inositol pyrophosphate metabolism using ip6k null amoeba

  7. Experimental Investigation of Mechanical and Thermal Properties of Silica Nanoparticle-Reinforced Poly(acrylamide) Nanocomposite Hydrogels.

    PubMed

    Zaragoza, Josergio; Babhadiashar, Nasim; O'Brien, Victor; Chang, Andrew; Blanco, Matthew; Zabalegui, Aitor; Lee, Hohyun; Asuri, Prashanth

    2015-01-01

    Current studies investigating properties of nanoparticle-reinforced polymers have shown that nanocomposites often exhibit improved properties compared to neat polymers. However, over two decades of research, using both experimental studies and modeling analyses, has not fully elucidated the mechanistic underpinnings behind these enhancements. Moreover, few studies have focused on developing an understanding among two or more polymer properties affected by incorporation of nanomaterials. In our study, we investigated the elastic and thermal properties of poly(acrylamide) hydrogels containing silica nanoparticles. Both nanoparticle concentration and size affected hydrogel properties, with similar trends in enhancements observed for elastic modulus and thermal diffusivity. We also observed significantly lower swellability for hydrogel nanocomposites relative to neat hydrogels, consistent with previous work suggesting that nanoparticles can mediate pseudo crosslinking within polymer networks. Collectively, these results indicate the ability to develop next-generation composite materials with enhanced mechanical and thermal properties by increasing the average crosslinking density using nanoparticles.

  8. Flocculation kinetics mechanism and floc formation prepared by poly aluminum chloride coupled with polyacrylamide for ship ballast water.

    PubMed

    Zhou, Zhimin; Liu, Sha; Jia, Linan

    2016-01-01

    The performance of flocculants prepared by poly aluminum chloride (PAC) and polyacrylamide (PAM) on treating ballast water collected at the Dalian new port area, the evaluation depending on the values of reaction parameters, and kinetics mechanism of flocculation were investigated in this study. Accordingly, the flocculants of 0.1 g·L(-1), prepared by mixing PAC of 10% with PAM of 2.0‰, enabled the removal rate of zooplankton and phytoplankton to reach 91% in ballast water at 20 °C. Based on flocculation kinetics mechanism analysis, the efficient vortex size during stirring should be larger than the floc particles, and gradient of fluctuating velocity provide the impetus for turbulence flocculation. The results of this study could be relevant to understanding particle-floc interactions during developmental flocculation, and during application of ballast water treatment.

  9. Microwave initiated synthesis of polyacrylamide grafted carboxymethylstarch (CMS-g-PAM): application as a novel matrix for sustained drug release.

    PubMed

    Sen, Gautam; Pal, Sagar

    2009-07-01

    This paper reports the investigation of microwave initiated synthesized polyacrylamide grafted carboxymethylstarch (CMS-g-PAM) as matrix for sustained drug release. 'In vitro' release of a model drug (5-amino salicylic acid) from CMS-g-PAM matrix has been studied. It is evident that higher the percentage grafting, more sustained is the rate of drug release. Further, the percentage grafting vs. t(50) value (i.e. time taken for release of 50% of the enclosed drug) correlation has been successfully studied for the first time. This correlation will lead to the possibility of a programmable drug release matrix based on grafted polysaccharide. In this matrix, the rate of release of the enclosed drug can be precisely programmed simply by adjustment of percentage grafting during synthesis.

  10. MAG2D: Interactive 2-1/2-dimensional magnetic modeling program (User's Guide and Documentation for Rev. 1)

    SciTech Connect

    Nutter, C.

    1981-04-01

    MAG2D is an interactive computer program used for modeling 2-1/2-dimensional magnetic data. A forward algorithm is used to give the theoretical attraction of magnetic intensity at a station due to a perturbing body given by the initial model. The resultant model can then be adjusted for a better fit by a combination of manual adjustment, one-dimensional automatic search, and Marquardt inversion. MAG2D has an interactive data management system for data manipulation and display built around subroutines to do a forward problem, a one-dimensional direct search and an inversion. These subroutines were originally separate batch-mode programs.

  11. The existence of energy solutions to 2-dimensional non-Lipschitz stochastic Navier-Stokes equations in unbounded domains

    NASA Astrophysics Data System (ADS)

    Taniguchi, Takeshi

    In this paper we consider the existence and uniqueness of weak energy solutions to a stochastic 2-dimensional non-Lipschitz Navier-Stokes equation perturbed by the cylindrical Wiener process W(t) in a bounded or unbounded domain D with the smooth boundary ∂ D or D=R: dX(t)=[-νAX(t)+B(X(t))] dt+f(t,X(t)) dt+g(t,X(t)) dW(t), where A is the Stokes operator and f, g satisfy the non-Lipschitz condition.

  12. Nanocrystal/sol-gel nanocomposites

    DOEpatents

    Klimov, Victor L.; Petruska, Melissa A.

    2010-05-25

    The present invention is directed to a process for preparing a solid composite having colloidal nanocrystals dispersed within a sol-gel matrix, the process including admixing colloidal nanocrystals with an amphiphilic polymer including hydrophilic groups selected from the group consisting of --COOH, --OH, --SO.sub.3H, --NH.sub.2, and --PO.sub.3H.sub.2 within a solvent to form an alcohol-soluble colloidal nanocrystal-polymer complex, admixing the alcohol-soluble colloidal nanocrystal-polymer complex and a sol-gel precursor material, and, forming the solid composite from the admixture. The present invention is also directed to the resultant solid composites and to the alcohol-soluble colloidal nanocrystal-polymer complexes.

  13. Multiscale modeling of polyelectrolyte gels

    NASA Astrophysics Data System (ADS)

    Wallmersperger, Thomas; Wittel, Falk K.; Kröplin, Bernd H.

    2006-03-01

    Electrolyte polymer gels are a very attractive class of actuation materials with remarkable electronic and mechanical properties having a great similarity to biological contractile tissues. They consist of a polymer network with ionizable groups and a liquid phase with mobile ions. Absorption and delivery of solvent lead to a considerably large change of volume. Due to this capability, they can be used as actuators for technical applications, where large swelling and shrinkage is desired. In the present work chemically and electrically stimulated polymer gels in a solution bath are investigated. To describe the different complicated phenomena occurring in these gels adequately, the modeling can be conducted on different scales. Therefore, models based on the statistical theory and porous media theory, as well as a multi-field model and a discrete element formulation are derived. A refinement of the different theories from global macroscopic to microscopic are presented in this paper: The statistical theory is a macroscopic theory capable to describe the global swelling or bending e.g. of a gel film, while the general theory of porous media (TPM) is a macroscopic continuum theory which is based on the theory of mixtures extended by the concept of volume fractions. The TPM is a homogenized model, i.e. all geometrical and physical quantities can be seen as statistical averages of the real quantities. The presented chemo-electro-mechanical multi-field formulation is a mesoscopic theory. It is capable of giving the concentrations and the electric potential in the whole domain. Finally the (micromechanical) discrete element (DE) theory is employed. In this case, the continuum is represented by distributed particles with local interaction relations combined with balance equations for the chemical field. This method is predestined for problems involving large displacements, strains and discontinuities. The presented formulations are compared and conclusions on their

  14. The gel electrophoresis markup language (GelML) from the Proteomics Standards Initiative.

    PubMed

    Gibson, Frank; Hoogland, Christine; Martinez-Bartolomé, Salvador; Medina-Aunon, J Alberto; Albar, Juan Pablo; Babnigg, Gyorgy; Wipat, Anil; Hermjakob, Henning; Almeida, Jonas S; Stanislaus, Romesh; Paton, Norman W; Jones, Andrew R

    2010-09-01

    The Human Proteome Organisation's Proteomics Standards Initiative has developed the GelML (gel electrophoresis markup language) data exchange format for representing gel electrophoresis experiments performed in proteomics investigations. The format closely follows the reporting guidelines for gel electrophoresis, which are part of the Minimum Information About a Proteomics Experiment (MIAPE) set of modules. GelML supports the capture of metadata (such as experimental protocols) and data (such as gel images) resulting from gel electrophoresis so that laboratories can be compliant with the MIAPE Gel Electrophoresis guidelines, while allowing such data sets to be exchanged or downloaded from public repositories. The format is sufficiently flexible to capture data from a broad range of experimental processes, and complements other PSI formats for MS data and the results of protein and peptide identifications to capture entire gel-based proteome workflows. GelML has resulted from the open standardisation process of PSI consisting of both public consultation and anonymous review of the specifications.

  15. One-step synthesis of highly efficient nanocatalysts on the supports with hierarchical pores using porous ionic liquid-water gel.

    PubMed

    Kang, Xinchen; Zhang, Jianling; Shang, Wenting; Wu, Tianbin; Zhang, Peng; Han, Buxing; Wu, Zhonghua; Mo, Guang; Xing, Xueqing

    2014-03-12

    Stable porous ionic liquid-water gel induced by inorganic salts was created for the first time. The porous gel was used to develop a one-step method to synthesize supported metal nanocatalysts. Au/SiO2, Ru/SiO2, Pd/Cu(2-pymo)2 metal-organic framework (Cu-MOF), and Au/polyacrylamide (PAM) were synthesized, in which the supports had hierarchical meso- and macropores, the size of the metal nanocatalysts could be very small (<1 nm), and the size distribution was very narrow even when the metal loading amount was as high as 8 wt %. The catalysts were extremely active, selective, and stable for oxidative esterification of benzyl alcohol to methyl benzoate, benzene hydrogenation to cyclohexane, and oxidation of benzyl alcohol to benzaldehyde because they combined the advantages of the nanocatalysts of small size and hierarchical porosity of the supports. In addition, this method is very simple.

  16. Rapid in vitro labeling procedures for two-dimensional gel fingerprinting

    SciTech Connect

    Lee, Y.F.; Fowlks, E.R.

    1982-01-15

    Improvements of existing in vitro procedures for labeling RNA radioactively, and modifications of the two-dimensional polyacrylamide gel electrophoresis system for making RNA fingerprints are described. These improvements are (a) inactivation of phosphatase with nitric acid at pH 2.0 eliminated the phenol-cholorform extraction step during 5'-end labeling with polynucleotide kinase and (..gamma..-/sup 32/P)ATP; (b) ZnSO/sub 4/ inactivation of R Nase T/sub 1/ results in a highly efficient procedure for 3'-end labeling with T4 ligase and (5'-/sup 32/P)pCp; and (c) a rapid 4-min procedure for variable quantity range of /sup 125/I and RNA results in a qualitative and quantitative sample for high-molecular weight RNA fingerprinting. Thus, these in vitro procedures become rapid and reproducible when combined with two-dimensional gel electrophoresis which eliminates simultaneously labeled impurities. Each labeling procedure is compared, using tobacco mosaic virus, Brome mosaic virus, and polio RNA. A series of Ap-rich oligonucleotides was discovered in the inner genome of Brome mosaic Virus RNA-3.

  17. Mathematical modelling of response of polymer gel dosimeters to brachytherapy radiation

    NASA Astrophysics Data System (ADS)

    Nasr, A. T.; Chain, J. N. M.; Schreiner, L. J.; McAuley, K. B.

    2010-11-01

    A dynamic partial differential equation (PDE) model is used to simulate effects of a single Ir192 brachytherapy seed on the amount and composition of polymer formed during polyacrylamide gel (PAG) dosimetry. Simulations are conducted for a point-source brachytherapy seed placed at the center of a 6%T 50% C anoxic PAG phantom. The seed is removed after one minute, but polymerization is simulated up to a final time of 24 hours. Simulation results indicate that changes occur in both the mass of polymer formed per unit dose and in the crosslink density as a function of the radial distance from the brachytherapy seed. For example, at a distance of 5 mm from the seed, 41 mg of polymer form per Gy of radiation absorbed (after 24 hours), whereas at a larger distance of 5 cm from the seed 75 mg of polymer form per Gy. The polymer that forms near the seed is predicted to have a higher level of crosslinking than the polymer that forms further away. These results suggest potential calibration problems that may occur during brachytherapy dosimetry using polymer gels.

  18. Development and evaluation of in situ gel-forming system for sustained delivery of insulin.

    PubMed

    Dhawan, Sanju; Kapil, Rishi; Kapoor, Deepak N

    2011-03-01

    Phase-sensitive in situ gel-forming controlled release formulations of insulin were prepared using poly(lactide-co-glycolide) and a solvent system consisting of various proportions of benzyl benzoate and benzyl alcohol. The in vitro release samples of formulations were assayed for insulin content by enzyme linked immunosorbent assay. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and circular dichroism spectroscopy studies of released insulin confirmed its conformational stability. The stability of insulin in the formulation was assessed using Fourier transform infrared spectroscopy. Rheological properties of the formulations, assessed under isothermal conditions, showed dilatant behavior of all the formulations. In vivo studies were carried out on the optimized formulations vis-à-vis pure insulin in mice and blood glucose levels were monitored for 15 days. Mean percentage reduction in blood glucose levels was calculated in all the animals and the results analyzed using ANOVA. The studies construed better pharmacodynamic response for the two optimized formulations in controlling the blood glucose levels vis-à-vis routine once-a-day administration of insulin. The subcutaneous tissues, further subjected to scanning electron microscopy studies and histopathological examinations, ascertained the biocompatibility of the formulation.

  19. Theoretical analysis of the kinetics of DNA hybridization with gel-immobilized oligonucleotides.

    PubMed Central

    Livshits, M A; Mirzabekov, A D

    1996-01-01

    A new method of DNA sequencing by hybridization using a microchip containing a set of immobilized oligonucleotides is being developed. A theoretical analysis is presented of the kinetics of DNA hybridization with deoxynucleotide molecules chemically tethered in a polyacrylamide gel layer. The analysis has shown that long-term evolution of the spatial distribution and of the amount of DNA bound in a hybridization cell is governed by "retarded diffusion," i.e., diffusion of the DNA interrupted by repeated association and dissociation with immobile oligonucleotide molecules. Retarded diffusion determines the characteristic time of establishing a final equilibrium state in a cell, i.e., the state with the maximum quantity and a uniform distribution of bound DNA. In the case of cells with the most stable, perfect duplexes, the characteristic time of retarded diffusion (which is proportional to the equilibrium binding constant and to the concentration of binding sites) can be longer than the duration of the real hybridization procedure. This conclusion is indirectly confirmed by the observation of nonuniform fluorescence of labeled DNA in perfect-match hybridization cells (brighter at the edges). For optimal discrimination of perfect duplexes from duplexes with mismatches the hybridization process should be brought to equilibrium under low-temperature nonsaturation conditions for all cells. The kinetic differences between perfect and nonperfect duplexes in the gel allow further improvement in the discrimination through additional washing at low temperature after hybridization. Images FIGURE 1 PMID:8913616

  20. Gel-based nonradioactive single-strand conformational polymorphism and mutation detection: limitations and solutions.

    PubMed

    Gupta, Vibhuti; Arora, Reetakshi; Gochhait, Sailesh; Bairwa, Narendra K; Bamezai, Rameshwar N K

    2014-01-01

    Single-strand conformation polymorphism (SSCP) for screening mutations/single-nucleotide polymorphisms (SNPs) is a simple, cost-effective technique, saving an expensive exercise of sequencing each and every polymerase chain reaction product and assisting in choosing only the amplicons of interest with expected mutations. The principle of detection of small changes in DNA sequences is based on changes in single-strand DNA conformations. The changes in electrophoretic mobility that SSCP detects are sequence dependent. The limitations faced in SSCP range from routine polyacrylamide gel electrophoresis (PAGE) problems to the problems of resolving mutant DNA bands. Both these problems can be solved by controlling PAGE conditions and by varying physical and environmental conditions such as pH, temperature, voltage, gel type and percentage, addition of additives or denaturants, and others. Despite much upgrading of the technology for mutation detection, SSCP remains the method of choice to analyze mutations and SNPs in order to understand genomic variations, both spontaneous and induced, and the genetic basis of diseases.

  1. Quantitative proteomics: assessing the spectrum of in-gel protein detection methods

    PubMed Central

    Gauci, Victoria J.; Wright, Elise P.

    2010-01-01

    Proteomics research relies heavily on visualization methods for detection of proteins separated by polyacrylamide gel electrophoresis. Commonly used staining approaches involve colorimetric dyes such as Coomassie Brilliant Blue, fluorescent dyes including Sypro Ruby, newly developed reactive fluorophores, as well as a plethora of others. The most desired characteristic in selecting one stain over another is sensitivity, but this is far from the only important parameter. This review evaluates protein detection methods in terms of their quantitative attributes, including limit of detection (i.e., sensitivity), linear dynamic range, inter-protein variability, capacity for spot detection after 2D gel electrophoresis, and compatibility with subsequent mass spectrometric analyses. Unfortunately, many of these quantitative criteria are not routinely or consistently addressed by most of the studies published to date. We would urge more rigorous routine characterization of stains and detection methodologies as a critical approach to systematically improving these critically important tools for quantitative proteomics. In addition, substantial improvements in detection technology, particularly over the last decade or so, emphasize the need to consider renewed characterization of existing stains; the quantitative stains we need, or at least the chemistries required for their future development, may well already exist. PMID:21686332

  2. Comparison of two gel filtration chromatographic methods for the purification of Lily symptomless virus.

    PubMed

    Wang, Ruoyu; Wang, Jianhui; Li, Juan; Wang, Yun; Xie, Zhongkui; An, Lizhe

    2007-02-01

    Lily symptomless virus (LSV) occurs frequently in many Lilium species worldwide and often causes developmental abnormalities such as a smaller flower and lower bulb yield. In this study, two moderate and efficient gel filtration chromatography (GFC) methods was compared, these two techniques were, respectively, based on Superdex-200 HR and Sephacryl S-1000 SF. The products purified by the two methods were then characterized by measurements with UV-spectrophotometer, reverse transcriptase (RT)-PCR, transmission electron microscope (TEM), polyacrylamide gel electrophoresis (PAGE), Western blotting and matrix assisted laser desorption-ionisation/time-of-flight mass spectrometry (MALDI-TOF-MS). The final yield of purified LSV by the Superdex-200 HR GFC was 9.4 mg from 50 g of fresh infected tissues of Lanzhou lily. However, from the same amount tissues, only 5.6 mg of LSV were obtained by using Sephacryl S-1000 SF GFC. The Superdex-200 HR method was thus shown to be more suitable for the purification of LSV than the Sephacryl S-1000 SF GFC. The Superdex-200 HR method does not require costly equipment for density centrifugation and ultracentrifugation. Furthermore, it can provide an economical and efficient way to obtain purified products for the preparation of antibodies for serological diagnosis or LSV infection and related investigations.

  3. A tunable isoelectric focusing via moving reaction boundary for two-dimensional gel electrophoresis and proteomics.

    PubMed

    Guo, Chen-Gang; Shang, Zhi; Yan, Jian; Li, Si; Li, Guo-Qing; Liu, Rong-Zhong; Qing, Ying; Fan, Liu-Yin; Xiao, Hua; Cao, Cheng-Xi

    2015-05-01

    Routine native immobilized pH gradient isoelectric focusing (IPG-IEF) and two-dimensional gel electrophoresis (2DE) are still suffering from unfortunate reproducibility, poor resolution (caused by protein precipitation) and instability in characterization of intact protein isoforms and posttranslational modifications. Based on the concept of moving reaction boundary (MRB), we firstly proposed a tunable non-IPG-IEF system to address these issues. By choosing proper pairs of catholyte and anolyte, we could achieve desired cathodic and anodic migrating pH gradients in non-IPG-IEF system, effectively eliminating protein precipitation and uncertainty of quantitation existing in routine IEF and 2DE, and enhancing the resolution and sensitivity of IEF. Then, an adjustable 2DE system was developed by combining non-IPG-IEF with polyacrylamide gel electrophoresis (PAGE). The improved 2DE was evaluated by testing model proteins and colon cancer cell lysates. The experiments revealed that (i) a tunable pH gradient could be designed via MRB; (ii) up to 1.65 fold improvement of resolution was achieved via non-IPG-IEF; (iii) the sensitivity of developed techniques was increased up to 2.7 folds; and (iv) up to about 16.4% more protein spots could be observed via the adjustable 2DE as compared with routine one. The developed techniques might contribute to complex proteome research, especially for screening of biological marker and analysis of extreme acidic/alkaline proteins.

  4. Radiation-induced refraction artifacts in the optical CT readout of polymer gel dosimeters

    SciTech Connect

    Campbell, Warren G.; Jirasek, Andrew; Wells, Derek M.

    2014-11-01

    Purpose: The objective of this work is to demonstrate imaging artifacts that can occur during the optical computed tomography (CT) scanning of polymer gel dosimeters due to radiation-induced refractive index (RI) changes in polyacrylamide gels. Methods: A 1 L cylindrical polyacrylamide gel dosimeter was irradiated with 3 × 3 cm{sup 2} square beams of 6 MV photons. A prototype fan-beam optical CT scanner was used to image the dosimeter. Investigative optical CT scans were performed to examine two types of rayline bending: (i) bending within the plane of the fan-beam and (ii) bending out the plane of the fan-beam. To address structured errors, an iterative Savitzky–Golay (ISG) filtering routine was designed to filter 2D projections in sinogram space. For comparison, 2D projections were alternatively filtered using an adaptive-mean (AM) filter. Results: In-plane rayline bending was most notably observed in optical CT projections where rays of the fan-beam confronted a sustained dose gradient that was perpendicular to their trajectory but within the fan-beam plane. These errors caused distinct streaking artifacts in image reconstructions due to the refraction of higher intensity rays toward more opaque regions of the dosimeter. Out-of-plane rayline bending was observed in slices of the dosimeter that featured dose gradients perpendicular to the plane of the fan-beam. These errors caused widespread, severe overestimations of dose in image reconstructions due to the higher-than-actual opacity that is perceived by the scanner when light is bent off of the detector array. The ISG filtering routine outperformed AM filtering for both in-plane and out-of-plane rayline errors caused by radiation-induced RI changes. For in-plane rayline errors, streaks in an irradiated region (>7 Gy) were as high as 49% for unfiltered data, 14% for AM, and 6% for ISG. For out-of-plane rayline errors, overestimations of dose in a low-dose region (∼50 cGy) were as high as 13 Gy for

  5. Identification of a Zn(2+)-sensitive component of Ehrlich cell plasma membrane redox system by CHAPS-agarose-polyacrylamide electrophoresis and in situ staining of activity.

    PubMed

    Rodríguez-Caso, L; Rodríguez-Agudo, D; del Castillo-Olivares, A; Márquez, J; Núñez de Castro, I; Medina, M A

    1997-01-01

    A procedure based on CHAPS-agarose-polyacrylamide electrophoresis and in situ staining of activity was used to detect a Zn(2+)-sensitive component of Ehrlich cell plasma membrane redox system. The procedure is so powerful that it allows to use crude plasma membrane fractions and can be easily adapted for use in an electrophoretic approach to the purification of this protein.

  6. Accretions of dark matter and dark energy onto (n+2)-dimensional Schwarzschild black hole and Morris-Thorne wormhole

    NASA Astrophysics Data System (ADS)

    Debnath, Ujjal

    2015-12-01

    In this work, we have studied accretion of the dark matter and dark energy onto of (n+2)-dimensional Schwarzschild black hole and Morris-Thorne wormhole. The mass and the rate of change of mass for (n+2)-dimensional Schwarzschild black hole and Morris-Thorne wormhole have been found. We have assumed some candidates of dark energy like holographic dark energy, new agegraphic dark energy, quintessence, tachyon, DBI-essence, etc. The black hole mass and the wormhole mass have been calculated in term of redshift when dark matter and above types of dark energies accrete onto them separately. We have shown that the black hole mass increases and wormhole mass decreases for holographic dark energy, new agegraphic dark energy, quintessence, tachyon accretion and the slope of increasing/decreasing of mass sensitively depends on the dimension. But for DBI-essence accretion, the black hole mass first increases and then decreases and the wormhole mass first decreases and then increases and the slope of increasing/decreasing of mass not sensitively depends on the dimension.

  7. Metal-silica sol-gel materials

    NASA Technical Reports Server (NTRS)

    Stiegman, Albert E. (Inventor)

    2002-01-01

    The present invention relates to a single phase metal-silica sol-gel glass formed by the co-condensation of a transition metal with silicon atoms where the metal atoms are uniformly distributed within the sol-gel glass as individual metal centers. Any transition metal may be used in the sol-gel glasses. The present invention also relates to sensor materials where the sensor material is formed using the single phase metal-silica sol-gel glasses. The sensor materials may be in the form of a thin film or may be attached to an optical fiber. The present invention also relates to a method of sensing chemicals using the chemical sensors by monitoring the chromatic change of the metal-silica sol-gel glass when the chemical binds to the sensor. The present invention also relates to oxidation catalysts where a metal-silica sol-gel glass catalyzes the reaction. The present invention also relates to a method of performing oxidation reactions using the metal-silica sol-gel glasses. The present invention also relates to organopolymer metal-silica sol-gel composites where the pores of the metal-silica sol-gel glasses are filled with an organic polymer polymerized by the sol-gel glass.

  8. Molecular imprinting in sol-gel matrix.

    PubMed

    Gupta, Radha; Kumar, Ashok

    2008-01-01

    Molecular imprinting is a newly developed methodology which provides molecular assemblies of desired structures and properties and is being increasingly used for several applications such as in separation processes, microreactors, immunoassays and antibody mimics, catalysis, artificial enzymes, biosensor recognition elements and bio- and chemo-sensors. The ambient processing conditions and versatility of the sol-gel process makes sol-gel glassy matrix suitable for molecular imprinting. The progress of sol-gel based molecular imprinted polymers (MIPs) for various applications can be seen from the growing number of publications. The main focus of the review is molecular imprinting in sol-gel matrix and applications of molecular imprinted sol-gel derived materials for the development of sensors. Combining sol-gel process with molecular imprinting enables to procure the sensors with greater sensitivity and selectivity necessary for sensing applications. The merits, problems, challenges and factors affecting molecular imprinting in sol-gel matrix have been discussed. Considerable attention has been drawn on recent developments like use of organically modified silane precursors (ORMOSILS) for the synthesis of hybrid molecular imprinted polymers (HMIPs) and applying surface sol-gel process for molecular imprinting. The development of molecular imprinted sol-gel nanotubes for biochemical separation and bio-imprinting is a new advancement and is under progress. Templated xerogels and molecularly imprinted sol-gel films provide a good platform for various sensor applications.

  9. Adapting capillary gel electrophoresis as a sensitive, high-throughput method to accelerate characterization of nucleic acid metabolic enzymes.

    PubMed

    Greenough, Lucia; Schermerhorn, Kelly M; Mazzola, Laurie; Bybee, Joanna; Rivizzigno, Danielle; Cantin, Elizabeth; Slatko, Barton E; Gardner, Andrew F

    2016-01-29

    Detailed biochemical characterization of nucleic acid enzymes is fundamental to understanding nucleic acid metabolism, genome replication and repair. We report the development of a rapid, high-throughput fluorescence capillary gel electrophoresis method as an alternative to traditional polyacrylamide gel electrophoresis to characterize nucleic acid metabolic enzymes. The principles of assay design described here can be applied to nearly any enzyme system that acts on a fluorescently labeled oligonucleotide substrate. Herein, we describe several assays using this core capillary gel electrophoresis methodology to accelerate study of nucleic acid enzymes. First, assays were designed to examine DNA polymerase activities including nucleotide incorporation kinetics, strand displacement synthesis and 3'-5' exonuclease activity. Next, DNA repair activities of DNA ligase, flap endonuclease and RNase H2 were monitored. In addition, a multicolor assay that uses four different fluorescently labeled substrates in a single reaction was implemented to characterize GAN nuclease specificity. Finally, a dual-color fluorescence assay to monitor coupled enzyme reactions during Okazaki fragment maturation is described. These assays serve as a template to guide further technical development for enzyme characterization or nucleoside and non-nucleoside inhibitor screening in a high-throughput manner.

  10. Adapting capillary gel electrophoresis as a sensitive, high-throughput method to accelerate characterization of nucleic acid metabolic enzymes

    PubMed Central

    Greenough, Lucia; Schermerhorn, Kelly M.; Mazzola, Laurie; Bybee, Joanna; Rivizzigno, Danielle; Cantin, Elizabeth; Slatko, Barton E.; Gardner, Andrew F.

    2016-01-01

    Detailed biochemical characterization of nucleic acid enzymes is fundamental to understanding nucleic acid metabolism, genome replication and repair. We report the development of a rapid, high-throughput fluorescence capillary gel electrophoresis method as an alternative to traditional polyacrylamide gel electrophoresis to characterize nucleic acid metabolic enzymes. The principles of assay design described here can be applied to nearly any enzyme system that acts on a fluorescently labeled oligonucleotide substrate. Herein, we describe several assays using this core capillary gel electrophoresis methodology to accelerate study of nucleic acid enzymes. First, assays were designed to examine DNA polymerase activities including nucleotide incorporation kinetics, strand displacement synthesis and 3′-5′ exonuclease activity. Next, DNA repair activities of DNA ligase, flap endonuclease and RNase H2 were monitored. In addition, a multicolor assay that uses four different fluorescently labeled substrates in a single reaction was implemented to characterize GAN nuclease specificity. Finally, a dual-color fluorescence assay to monitor coupled enzyme reactions during Okazaki fragment maturation is described. These assays serve as a template to guide further technical development for enzyme characterization or nucleoside and non-nucleoside inhibitor screening in a high-throughput manner. PMID:26365239

  11. Separation and identification of Musa acuminate Colla (banana) leaf proteins by two-dimensional gel electrophoresis and mass spectrometry.

    PubMed

    Lu, Y; Qi, Y X; Zhang, H; Zhang, H Q; Pu, J J; Xie, Y X

    2013-12-19

    To establish a proteomic reference map of Musa acuminate Colla (banana) leaf, we separated and identified leaf proteins using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and mass spectrometry (MS). Tryptic digests of 44 spots were subjected to peptide mass fingerprinting (PMF) by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS. Three spots that were not identified by MALDI-TOF MS analysis were identified by searching against the NCBInr, SwissProt, and expressed sequence tag (EST) databases. We identified 41 unique proteins. The majority of the identified leaf proteins were found to be involved in energy metabolism. The results indicate that 2D-PAGE is a sensitive and powerful technique for the separation and identification of Musa leaf proteins. A summary of the identified proteins and their putative functions is discussed.

  12. Photorefractive sol-gel materials

    SciTech Connect

    Chaput, F.; Boilot, J.P.; Gacoin, T.; Darracq, B.; Riehl, D.; Canva, M.; Levy, Y.; Brun, A.

    1996-12-31

    The authors report the synthesis and characterization of photorefractive sol-gel materials that possess covalently attached push-pull azobenzene and carbazole moieties. Molecular structural characterization of the modified silane monomers was achieved by {sup 1}H NMR and infra red spectroscopy. The second-order nonlinear optical properties of the organic-inorganic hybrid films prepared from modified silane monomers were evaluated by second-harmonic generation. The stabilized value of the second harmonic coefficient, d{sub 33}, of films poled by corona discharge, at 1,064 nm fundamental wavelength was found to be 107 pm/V. Photorefractivity was clearly displayed from a two beam coupling experiment.

  13. Proteomic Profiling of Rat Thyroarytenoid Muscle

    ERIC Educational Resources Information Center

    Welham, Nathan V.; Marriott, Gerard; Bless, Diane M.

    2006-01-01

    Purpose: Proteomic methodologies offer promise in elucidating the systemwide cellular and molecular processes that characterize normal and diseased thyroarytenoid (TA) muscle. This study examined methodological issues central to the application of 2-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (2D SDS-PAGE) to the study of…

  14. Superoxide dismutase isozyme detection using two-dimensional gel electrophoresis zymograms.

    PubMed

    Niyomploy, Ploypat; Srisomsap, Chantragan; Chokchaichamnankit, Daranee; Vinayavekhin, Nawaporn; Karnchanatat, Aphichart; Sangvanich, Polkit

    2014-03-01

    Superoxide dismutases (SODs) are ubiquitous antioxidant enzymes involved in cell protection from reactive oxygen species. Their antioxidant activities make them of interest to applied biotechnology industries and are usually sourced from plants. SODs are also involved in stress signaling responses in plants, and can be used as indicators of these responses. In this article, a suitable method for the separation of different SOD isoforms using two-dimensional-gel electrophoresis (2D-GE) zymograms is reported. The method was developed with a SOD standard from bovine erythrocytes and later applied to extracts from Stemona tuberosa. The first (non-denaturing isoelectric focusing) and second (denaturing sodium dodecylsulphate-polyacrylamide gel electrophoresis) dimensions of duplicate 2D-GE gels were stained with either Coomassie brilliant blue G-250 for total protein visualization, or SOD activity (zymogram) using riboflavin/nitroblue tetrazolium. For confirmation, putative SOD activity positive spots were subject to trypsin digestion and nano-liquid chromatography tandem mass spectrometry, followed by searching the MASCOT database for potential identification. The method could separate different SOD isoforms from a plant extract and at least partially maintain or allow renaturation to the native forms of the enzyme. Peptide sequencing of the 2D-GE suggested that the SODs were resolved correctly, identifying the control CuZn-SOD from bovine erythrocytes. The two SODs from S. tuberosa tubers were found to be likely homologous of a CuZn-SOD. SOD detection and isoform separation by 2D-GE zymograms was efficient and reliable. The method is likely applicable to SOD detection from plants or other organisms. Moreover, a similar approach could be developed for detection of other important enzymes in the future.

  15. Mucin Agarose Gel Electrophoresis: Western Blotting for High-molecular-weight Glycoproteins.

    PubMed

    Ramsey, Kathryn A; Rushton, Zachary L; Ehre, Camille

    2016-06-14

    Mucins, the heavily-glycosylated proteins lining mucosal surfaces, have evolved as a key component of innate defense by protecting the epithelium against invading pathogens. The main role of these macromolecules is to facilitate particle trapping and clearance while promoting lubrication of the mucosa. During protein synthesis, mucins undergo intense O-glycosylation and multimerization, which dramatically increase the mass and size of these molecules. These post-translational modifications are critical for the viscoelastic properties of mucus. As a result of the complex biochemical and biophysical nature of these molecules, working with mucins provides many challenges that cannot be overcome by conventional protein analysis methods. For instance, their high-molecular-weight prevents electrophoretic migration via regular polyacrylamide gels and their sticky nature causes adhesion to experimental tubing. However, investigating the role of mucins in health (e.g., maintaining mucosal integrity) and disease (e.g., hyperconcentration, mucostasis, cancer) has recently gained interest and mucins are being investigated as a therapeutic target. A better understanding of the production and function of mucin macromolecules may lead to novel pharmaceutical approaches, e.g., inhibitors of mucin granule exocytosis and/or mucolytic agents. Therefore, consistent and reliable protocols to investigate mucin biology are critical for scientific advancement. Here, we describe conventional methods to separate mucin macromolecules by electrophoresis using an agarose gel, transfer protein into nitrocellulose membrane, and detect signal with mucin-specific antibodies as well as infrared fluorescent gel reader. These techniques are widely applicable to determine mucin quantitation, multimerization and to test the effects of pharmacological compounds on mucins.

  16. Active gel model of amoeboid cell motility

    NASA Astrophysics Data System (ADS)

    Callan-Jones, A. C.; Voituriez, R.

    2013-02-01

    We develop a model of amoeboid cell motility based on active gel theory. Modeling the motile apparatus of a eukaryotic cell as a confined layer of finite length of poroelastic active gel permeated by a solvent, we first show that, due to active stress and gel turnover, an initially static and homogeneous layer can undergo a contractile-type instability to a polarized moving state in which the rear is enriched in gel polymer. This agrees qualitatively with motile cells containing an actomyosin-rich uropod at their rear. We find that the gel layer settles into a steadily moving, inhomogeneous state at long times, sustained by a balance between contractility and filament turnover. In addition, our model predicts an optimal value of the gel-substrate adhesion leading to maximum layer speed, in agreement with cell motility assays. The model may be relevant to motility of cells translocating in complex, confining environments that can be mimicked experimentally by cell migration through microchannels.

  17. Automated apparatus for producing gradient gels

    DOEpatents

    Anderson, N.L.

    1983-11-10

    Apparatus for producing a gradient gel which serves as a standard medium for a two-dimensional analysis of proteins, the gel having a density gradient along its height formed by a variation in gel composition, with the apparatus including first and second pumping means each including a plurality of pumps on a common shaft and driven by a stepping motor capable of providing small incremental changes in pump outputs for the gel ingredients, the motors being controlled, by digital signals from a digital computer, a hollow form or cassette for receiving the gel composition, means for transferring the gel composition including a filler tube extending near the bottom of the cassette, adjustable horizontal and vertical arms for automatically removing and relocating the filler tube in the next cassette, and a digital computer programmed to automatically control the stepping motors, arm movements, and associated sensing operations involving the filling operation.

  18. Automated apparatus for producing gradient gels

    DOEpatents

    Anderson, Norman L.

    1986-01-01

    Apparatus for producing a gradient gel which serves as a standard medium for a two-dimensional analysis of proteins, the gel having a density gradient along its height formed by a variation in gel composition, with the apparatus including first and second pumping means each including a plurality of pumps on a common shaft and driven by a stepping motor capable of providing small incremental changes in pump outputs for the gel ingredients, the motors being controlled, by digital signals from a digital computer, a hollow form or cassette for receiving the gel composition, means for transferring the gel composition including a filler tube extending near the bottom of the cassette, adjustable horizontal and vertical arms for automatically removing and relocating the filler tube in the next cassette, and a digital computer programmed to automatically control the stepping motors, arm movements, and associated sensing operations involving the filling operation.

  19. Self-Pumping Active Gel

    NASA Astrophysics Data System (ADS)

    Wu, Kun-Ta; Hishamunda, Jean Bernard; Fraden, Seth; Dogic, Zvonimir

    Isotropic active gels are the network which is consist of cross-linked building blocks and the structure of which changes randomly and isotropically with time. Dogic et. al. show that pairs of anti-parallel microtubules form extensile bundles, which merge, extend, and buckle. In an unconfined system, the dynamics of these bundles causes spontaneous turbulent-like flow driven by motion of microscopic molecular motors. We found that confining these active gels in a millimeter sized toroids causes a transition into a new dynamical state characterized by circulation currents persisting for hours until ATP is depleted. We show how toroid dimensions impact the properties of self-organized circular currents, how directions of circulation can be designed by engineering ratchet-shaped boundaries, and how circulations of connected toroids can be either synchronized or antisynchronized. Furthermore, we demonstrate that the flow rate in the circulation is independent of curvature and length of flow path. The flow rate persists for centimeters without decay, disregarding conventional pipe flow resistance. Such findings pave the path to self-pumping pipe transport and performing physical work with biological system.

  20. Gel trapping of dense colloids.

    PubMed

    Laxton, Peter B; Berg, John C

    2005-05-01

    Phase density differences in sols, foams, or emulsions often lead to sedimentation or creaming, causing problems for materials where spatial uniformity over extended periods of time is essential. The problem may be addressed through the use of rheology modifiers in the continuous phase. Weak polymer gels have found use for this purpose in the food industry where they appear to be capable of trapping dispersoid particles in a three-dimensional matrix while displaying water-like viscosities at low shear. Attempts to predict sedimentation stability in terms of particle properties (size, shape, density difference) and gel yield stress have led to qualitative success for suspensions of large particles. The effect of particle size, however, in particular the case in which colloidal dimensions are approached, has not been investigated. The present work seeks to determine useful stability criteria for colloidal dispersions in terms of readily accessible viscoelastic descriptors. Results are reported for systems consisting of 12 microm poly(methyl methacrylate) (PMMA) spheres dispersed in aqueous gellan gum. Monovalent salt concentration is varied to control rheological properties, and sedimentation/centrifugation experiments are performed to determine dispersion stability. Necessary conditions for stability consist of a minimum yield stress together with a value of tan delta less than unity.