Sample records for zwitterionic detergent chaps

  1. Comparative study of the interaction of CHAPS and Triton X-100 with the erythrocyte membrane.

    PubMed

    Rodi, P M; Bocco Gianello, M D; Corregido, M C; Gennaro, A M

    2014-03-01

    The zwitterionic detergent CHAPS, a derivative of the bile salts, is widely used in membrane protein solubilization. It is a "facial" detergent, having a hydrophilic side and a hydrophobic back. The objective of this work is to characterize the interaction of CHAPS with a cell membrane. To this aim, erythrocytes were incubated with a wide range of detergent concentrations in order to determine CHAPS partition behavior, and its effects on membrane lipid order, hemolytic effects, and the solubilization of membrane phospholipids and cholesterol. The results were compared with those obtained with the nonionic detergent Triton X-100. It was found that CHAPS has a low affinity for the erythrocyte membrane (partition coefficient K=0.06mM(-1)), and at sub-hemolytic concentrations it causes little effect on membrane lipid order. CHAPS hemolysis and phospholipid solubilization are closely correlated. On the other side, binding of Triton X-100 disorders the membrane at all levels, and has independent mechanisms for hemolysis and solubilization. Differential behavior was observed in the solubilization of phospholipids and cholesterol. Thus, the detergent resistant membranes (DRM) obtained with the two detergents will have different composition. The behaviors of the two detergents are related to the differences in their molecular structures, suggesting that CHAPS does not penetrate the lipid bilayer but binds in a flat position on the erythrocyte surface, both in intact and cholesterol depleted erythrocytes. A relevant result for Triton X-100 is that hemolysis is not directly correlated with the solubilization of membrane lipids, as it is usually assumed. Copyright © 2013 Elsevier B.V. All rights reserved.

  2. Stabilization of photosystem II reaction centers: influence of bile salt detergents and low pH.

    PubMed

    Gall, B; Scheer, H

    1998-07-17

    Rapid deterioration of samples is a major obstacle in research on the isolated reaction center of photosystem II. Its stability was tested systematically using a wide range of detergents, varying pH and temperature. Stability and activity did not depend on ionic properties of detergents or on critical micellar concentration. However, both were significantly increased by bile salt detergents in the dark as well as in the light. Low pH (5.5) and low temperature further improved stability. The results suggest that in particular the zwitterionic bile salt detergent, CHAPS, in pH 5.5 buffers is a very useful detergent and even superior to dodecylmaltoside for work with photosystem II reaction centers.

  3. Efficient production and purification of functional bacteriorhodopsin with a wheat-germ cell-free system and a combination of Fos-choline and CHAPS detergents.

    PubMed

    Genji, Takahisa; Nozawa, Akira; Tozawa, Yuzuru

    2010-10-01

    Cell-free translation is one potential approach to the production of functional transmembrane proteins. We have now examined various detergents as supplements to a wheat-germ cell-free system in order to optimize the production and subsequent purification of a functional model transmembrane protein, bacteriorhodopsin. We found that Fos-choline and CHAPS detergents counteracted each other's inhibitory effects on cell-free translation activity and thereby allowed the efficient production and subsequent purification of functional bacteriorhodopsin in high yield. Copyright © 2010 Elsevier Inc. All rights reserved.

  4. Dependence of the product chain-length on detergents for long-chain E-polyprenyl diphosphate synthases

    PubMed Central

    Pan, Jian-Jung; Ramamoorthy, Gurusankar; Poulter, C. Dale

    2013-01-01

    Long-chain E-polyprenyl diphosphate synthases (E-PDS) catalyze repetitive addition of isopentenyl diphosphate (IPP) to the growing prenyl chain of an allylic diphosphate. The polyprenyl diphosphate products are required for the biosynthesis of ubiquinones and menaquinones required for electron transport during oxidative phosphorylation to generate ATP. In vitro, the long-chain PDSs require addition of phospholipids or detergents to the assay buffer to enhance product release and maintain efficient turnover. During preliminary assays of product chain-length with anionic, zwitterionic, and non-ionic detergents, we discovered considerable variability. Examination of a series of non-ionic PEG detergents with several long-chain E-PDSs from different organisms revealed that in vitro incubations with nonaethylene glycol monododecyl ether or Triton X-100 typically gave chain lengths that corresponded to those of the isoprenoid moieties in respiratory quinones synthesized in vivo. In contrast incubations in buffer with n-butanol, CHAPS, DMSO, n-octyl-β-glucopyranoside, or β-cyclodextrin or in buffer without detergent typically proceeded more slowly and gave a broad range of chain lengths. PMID:23802587

  5. Identification of detergent-resistant plasma membrane microdomains in dictyostelium: enrichment of signal transduction proteins.

    PubMed

    Xiao, Z; Devreotes, P N

    1997-05-01

    Unlike most other cellular proteins, the chemoattractant receptor, cAR1, of Dictyostelium is resistant to extraction by the zwitterionic detergent, CHAPS. We exploited this property to isolate a subcellular fraction highly enriched in cAR1 by flotation of CHAPS lysates of cells in sucrose density gradients. Immunogold electron microscopy studies revealed a homogeneous preparation of membrane bilayer sheets. This preparation, designated CHAPS-insoluble floating fraction (CHIEF), also contained a defined set of 20 other proteins and a single uncharged lipid. Cell surface biotinylation and preembedding immunoelectron microscopy both confirmed the plasma membrane origin of this preparation. The cell surface phosphodiesterase (PDE) and a downstream effector of cAR1, adenylate cyclase (ACA), were specifically localized in these structures, whereas the cell adhesion molecule gp80, most of the major cell surface membrane proteins, cytoskeletal components, the actin-binding integral membrane protein ponticulin, and G-protein alpha- and beta-subunits were absent. Overall, CHIFF represents about 3-5% of cell externally exposed membrane proteins. All of these results indicate that CHIFF is derived from specialized microdomains of the plasma membrane. The method of isolation is analogous to that of caveolae. However, we were unable to detect distinct caveolae-like structures on the cell surface associated with cAR1, which showed a diffuse staining profile. The discovery of CHIFF facilitates the purification of cAR1 and related signaling proteins and the biochemical characterization of receptor-mediated processes such as G-protein activation and desensitization. It also has important implications for the "fluid mosaic" model of the plasma membrane structures.

  6. Crystal structure of the EnvZ periplasmic domain with CHAPS.

    PubMed

    Hwang, Eunha; Cheong, Hae-Kap; Kim, Sang-Yoon; Kwon, Ohsuk; Blain, Katherine Y; Choe, Senyon; Yeo, Kwon Joo; Jung, Yong Woo; Jeon, Young Ho; Cheong, Chaejoon

    2017-05-01

    Bacteria sense and respond to osmolarity through the EnvZ-OmpR two-component system. The structure of the periplasmic sensor domain of EnvZ (EnvZ-PD) is not available yet. Here, we present the crystal structure of EnvZ-PD in the presence of CHAPS detergent. The structure of EnvZ-PD shows similar folding topology to the PDC domains of PhoQ, DcuS, and CitA, but distinct orientations of helices and β-hairpin structures. The CD and NMR spectra of EnvZ-PD in the presence of cholate, a major component of bile salts, are similar to those with CHAPS. Chemical cross-linking shows that the dimerization of EnvZ-PD is significantly inhibited by the CHAPS and cholate. Together with β-galactosidase assay, these results suggest that bile salts may affect the EnvZ structure and function in Escherichia coli. © 2017 Federation of European Biochemical Societies.

  7. Impact of purification conditions and history on A 2A adenosine receptor activity: The role of CHAPS and lipids

    DOE PAGES

    Naranjo, Andrea N.; McNeely, Patrick M.; Katsaras, John; ...

    2016-05-27

    The adenosine A 2A receptor (A 2AR) is a much-studied class A G protein-coupled receptor (GPCR). For biophysical studies, A 2AR is commonly purified in a detergent mixture of dodecylmaltoside (DDM), 3-(3-cholamidopropyl) dimethylammoniopropane sulfonate (CHAPS), and cholesteryl hemisuccinate (CHS). Here we studied the effects of CHAPS on the ligand binding activity and stability of wild type, full-length human A 2AR. We also tested the cholesterol requirement for maintaining the active conformation of the receptor when solubilized in detergent micelles. To this end, the receptor was purified using DDM, DDM/CHAPS, or the short hydrocarbon chain lipid 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC, di-6:0PC). After solubilizationmore » in DDM, DDM/CHAPS, or DHPC micelles, although A 2AR was found to retain its native-like fold, its binding ability was significantly compromised compared to DDM or DDM/CHAPS with CHS. It therefore appears that although cholesterol is not needed for A 2AR to retain a native-like, α-helical conformation, it may be a critical component for high affinity ligand binding. Further, this result suggests that the conformational differences between the active and inactive protein may be so subtle that commonly used spectroscopic methods are unable to differentiate between the two forms, highlighting the need for activity measurements. Furthermore, the studies presented in this paper also underline the importance of the protein’s purification history; i.e., detergents that interact with the protein during purification affect the ligand binding properties of the receptor in an irreversible manner.« less

  8. Impact of purification conditions and history on A 2A adenosine receptor activity: The role of CHAPS and lipids

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

    Naranjo, Andrea N.; McNeely, Patrick M.; Katsaras, John

    The adenosine A 2A receptor (A 2AR) is a much-studied class A G protein-coupled receptor (GPCR). For biophysical studies, A 2AR is commonly purified in a detergent mixture of dodecylmaltoside (DDM), 3-(3-cholamidopropyl) dimethylammoniopropane sulfonate (CHAPS), and cholesteryl hemisuccinate (CHS). Here we studied the effects of CHAPS on the ligand binding activity and stability of wild type, full-length human A 2AR. We also tested the cholesterol requirement for maintaining the active conformation of the receptor when solubilized in detergent micelles. To this end, the receptor was purified using DDM, DDM/CHAPS, or the short hydrocarbon chain lipid 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC, di-6:0PC). After solubilizationmore » in DDM, DDM/CHAPS, or DHPC micelles, although A 2AR was found to retain its native-like fold, its binding ability was significantly compromised compared to DDM or DDM/CHAPS with CHS. It therefore appears that although cholesterol is not needed for A 2AR to retain a native-like, α-helical conformation, it may be a critical component for high affinity ligand binding. Further, this result suggests that the conformational differences between the active and inactive protein may be so subtle that commonly used spectroscopic methods are unable to differentiate between the two forms, highlighting the need for activity measurements. Furthermore, the studies presented in this paper also underline the importance of the protein’s purification history; i.e., detergents that interact with the protein during purification affect the ligand binding properties of the receptor in an irreversible manner.« less

  9. Identification of detergent-resistant plasma membrane microdomains in dictyostelium: enrichment of signal transduction proteins.

    PubMed Central

    Xiao, Z; Devreotes, P N

    1997-01-01

    Unlike most other cellular proteins, the chemoattractant receptor, cAR1, of Dictyostelium is resistant to extraction by the zwitterionic detergent, CHAPS. We exploited this property to isolate a subcellular fraction highly enriched in cAR1 by flotation of CHAPS lysates of cells in sucrose density gradients. Immunogold electron microscopy studies revealed a homogeneous preparation of membrane bilayer sheets. This preparation, designated CHAPS-insoluble floating fraction (CHIEF), also contained a defined set of 20 other proteins and a single uncharged lipid. Cell surface biotinylation and preembedding immunoelectron microscopy both confirmed the plasma membrane origin of this preparation. The cell surface phosphodiesterase (PDE) and a downstream effector of cAR1, adenylate cyclase (ACA), were specifically localized in these structures, whereas the cell adhesion molecule gp80, most of the major cell surface membrane proteins, cytoskeletal components, the actin-binding integral membrane protein ponticulin, and G-protein alpha- and beta-subunits were absent. Overall, CHIFF represents about 3-5% of cell externally exposed membrane proteins. All of these results indicate that CHIFF is derived from specialized microdomains of the plasma membrane. The method of isolation is analogous to that of caveolae. However, we were unable to detect distinct caveolae-like structures on the cell surface associated with cAR1, which showed a diffuse staining profile. The discovery of CHIFF facilitates the purification of cAR1 and related signaling proteins and the biochemical characterization of receptor-mediated processes such as G-protein activation and desensitization. It also has important implications for the "fluid mosaic" model of the plasma membrane structures. Images PMID:9168471

  10. Soaping the NMDA receptor: Various types of detergents influence differently [(3)H]MK-801 binding to rat brain membranes.

    PubMed

    Berger, Michael L

    2016-01-01

    Membranes prepared from rat brain were treated with increasing concentrations of cationic, neutral, anionic and zwitterionic surfactants. Potent inactivation of [(3)H]MK-801 binding to NMDA receptors (NRs) was provided by the cation cetyl pyridinium (IC50 25 μM) and the neutral digitonin (IC50 37 μM). A 2 h incubation of rat brain membranes at 24°C with 100 μM of the neutral Triton X-100 resulted in about 50% reversible inhibition (without inactivation). Reversible inhibition was also effected by the anion deoxycholate (IC50 700 μM), and by the zwitterions N-lauryl sulfobetaine (12-SB(±), 400 μM) and CHAPS (1.5 mM), with inactivation at higher concentrations. Keeping the NR cation channel in the closed state significantly protected against inactivation by cations and by 12-SB(±), but not by the other detergents. Inactivation depended differentially on the amount of the membranes, on the duration of the treatment, and on the temperature. Varying the amount of membranes by a factor 8 yielded for cetyl trimethylammonium (16-NMe3(+)) IC50s of inactivation from 10 to 80 μM, while for deoxycholate the IC50 of inactivation was 1.2 mM for all tissue quantities. Some compounds inactivated within a few min (16-NMe3(+), digitonin, CHAPS), while inactivation by others took at least half an hour (Triton X-100, deoxycholate, 12-SB(±)). These last 3 ones also exhibited the steepest temperature dependence. Knowledge about the influence of various parameters is helpful in selecting appropriate conditions allowing the treatment of brain membranes with amphiphiles without risking irreversible inactivation. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Evaluation of eco-friendly zwitterionic detergents for enveloped virus inactivation.

    PubMed

    Conley, Lynn; Tao, Yinying; Henry, Alexis; Koepf, Edward; Cecchini, Douglas; Pieracci, John; Ghose, Sanchayita

    2017-04-01

    Inclusion of a detergent in protein biotherapeutic purification processes is a simple and very robust method for inactivating enveloped viruses. The detergent Triton X-100 has been used for many years and is part of the production process of several commercial therapeutic proteins. However, recent ecological studies have suggested that Triton X-100 and its break-down products can potentially behave as endocrine disrupters in aquatic organisms, raising concerns from an environmental impact perspective. As such, discharge of Triton X-100 into the waste water treatment plants is regulated in some jurisdictions, and alternative detergents for viral inactivation are required. In this work, we report on the identification and evaluation of more eco-friendly detergents as viable replacements for Triton X-100. Five detergent candidates with low to moderate environmental impact were initially identified and evaluated with respect to protein stability, followed by proof-of-concept virus inactivation studies using a model enveloped virus. From the set of candidates lauryldimethylamine N-oxide (LDAO) was identified as the most promising detergent due to its low ecotoxicity, robust anti-viral activity (LRV >4 at validation set-point conditions with X-MuLX), and absence of any negative impact on protein function. This detergent exhibited effective and robust virus inactivation in a broad range of protein concentrations, solution conductivities, pHs, and in several different cell culture fluid matrices. The only process parameter which correlated with reduced virus inactivation potency was LDAO concentration, and then only when the concentration was reduced to below the detergent's critical micelle concentration (CMC). Additionally, this work also demonstrated that LDAO was cleared to below detectable levels after Protein A affinity chromatography, making it suitable for use in a platform process that utilizes this chromatographic mode for protein capture. All these findings

  12. Sodium hydroxide based non-detergent decellularizing solution for rat lung.

    PubMed

    Sengyoku, Hideyori; Tsuchiya, Tomoshi; Obata, Tomohiro; Doi, Ryoichiro; Hashimoto, Yasumasa; Ishii, Mitsutoshi; Sakai, Hiromi; Matsuo, Naoto; Taniguchi, Daisuke; Suematsu, Takashi; Lawn, Murray; Matsumoto, Keitaro; Miyazaki, Takuro; Nagayasu, Takeshi

    2018-06-11

    Lung transplantation is the last option for the treatment of end stage chronic lung disorders. Because the shortage of donor lung organs represents the main hurdle, lung regeneration has been considered to overcome this hurdle. Recellularization of decellularized organ scaffold is a promising option for organ regeneration. Although detergents are ordinarily used for decellularization, other approaches are possible. Here we used high alkaline (pH12) sodium hydroxide (NaOH)-PBS solution without detergents for lung decellularization and compared the efficacy on DNA elimination and ECM preservation with detergent based decellularization solutions CHAPS and SDS. Immunohistochemical image analysis showed that cell components were removed by NaOH solution as well as other detergents. A Collagen and GAG assay showed that the collagen reduction of the NaOH group was comparable to that of the CHAPS and SDS groups. However, DNA reduction was more significant in the NaOH group than in other groups (p < 0.0001). The recellularization of HUVEC revealed cell attachment was not inferior to that of the SDS group. Ex vivo functional analysis showed 100% oxygen ventilation increased oxygen partial pressure as artificial hemoglobin vesicle-PBS solution passed through regenerated lungs in the SDS or NaOH group. It was concluded that the NaOH-PBS based decellularization solution was comparable to ordinal decellularizaton solutions and competitive in cost effectiveness and residues in the decellularized scaffold negligible, thus providing another potential option to detergent for future clinical usage.

  13. Membrane protein stability can be compromised by detergent interactions with the extramembranous soluble domains

    PubMed Central

    Yang, Zhengrong; Wang, Chi; Zhou, Qingxian; An, Jianli; Hildebrandt, Ellen; Aleksandrov, Luba A; Kappes, John C; DeLucas, Lawrence J; Riordan, John R; Urbatsch, Ina L; Hunt, John F; Brouillette, Christie G

    2014-01-01

    Detergent interaction with extramembranous soluble domains (ESDs) is not commonly considered an important determinant of integral membrane protein (IMP) behavior during purification and crystallization, even though ESDs contribute to the stability of many IMPs. Here we demonstrate that some generally nondenaturing detergents critically destabilize a model ESD, the first nucleotide-binding domain (NBD1) from the human cystic fibrosis transmembrane conductance regulator (CFTR), a model IMP. Notably, the detergents show equivalent trends in their influence on the stability of isolated NBD1 and full-length CFTR. We used differential scanning calorimetry (DSC) and circular dichroism (CD) spectroscopy to monitor changes in NBD1 stability and secondary structure, respectively, during titration with a series of detergents. Their effective harshness in these assays mirrors that widely accepted for their interaction with IMPs, i.e., anionic > zwitterionic > nonionic. It is noteworthy that including lipids or nonionic detergents is shown to mitigate detergent harshness, as will limiting contact time. We infer three thermodynamic mechanisms from the observed thermal destabilization by monomer or micelle: (i) binding to the unfolded state with no change in the native structure (all detergent classes); (ii) native state binding that alters thermodynamic properties and perhaps conformation (nonionic detergents); and (iii) detergent binding that directly leads to denaturation of the native state (anionic and zwitterionic). These results demonstrate that the accepted model for the harshness of detergents applies to their interaction with an ESD. It is concluded that destabilization of extramembranous soluble domains by specific detergents will influence the stability of some IMPs during purification. PMID:24652590

  14. Activation of pollen tube callose synthase by detergents. Evidence for different mechanisms of action.

    PubMed Central

    Li, H; Bacic, A; Read, S M

    1997-01-01

    In pollen tubes of Nicotiana alata, a membrane-bound, Ca(2+)-independent callose synthase (CalS) is responsible for the biosynthesis of the (1,3)-beta-glucan backbone of callose, the main cell wall component. Digitonin increases CalS activity 3- to 4-fold over a wide range of concentrations, increasing the maximum initial velocity without altering the Michaelis constant for UDP-glucose. The CalS activity that requires digitonin for assay (the latent CalS activity) is not inhibited by the membrane-impermeant, active site-directed reagent UDP-pyridoxal when the reaction is conducted in the absence of digitonin. This is consistent with digitonin increasing CalS activity by the permeabilization of membrane vesicles. A second group of detergents, including 3-[(3-cholamidopropyl)dimethylammonio]-1-propane-sulfonate (CHAPS), Zwittergent 3-16, and 1-alpha-lysolecithin, activate pollen tube CalS 10- to 15-fold, but only over a narrow range of concentrations just below their respective critical micellar concentrations. This activation could not be attributed to any particular chemical feature of these detergents. CHAPS increases maximum initial velocity and decreases the Michaelis constant for UDP-glucose and activates CalS even in the presence of permeabilizing concentrations of digitonin. Inhibition studies with UDP-pyridoxal indicate that activation by CHAPS occurs by recruitment of previously inactive CalS molecules to the pool of active enzyme. The activation of pollen tube CalS by these detergents therefore resembles activation of the enzyme by trypsin. PMID:9276948

  15. A Comparative Study of Rat Lung Decellularization by Chemical Detergents for Lung Tissue Engineering

    PubMed Central

    Tebyanian, Hamid; Karami, Ali; Motavallian, Ebrahim; Aslani, Jafar; Samadikuchaksaraei, Ali; Arjmand, Babak; Nourani, Mohammad Reza

    2017-01-01

    BACKGROUND: Lung disease is the most common cause of death in the world. The last stage of pulmonary diseases is lung transplantation. Limitation and shortage of donor organs cause to appear tissue engineering field. Decellularization is a hope for producing intact ECM in the development of engineered organs. AIM: The goal of the decellularization process is to remove cellular and nuclear material while retaining lung three-dimensional and molecular proteins. Different concentration of detergents was used for finding the best approach in lung decellularization. MATERIAL AND METHODS: In this study, three-time approaches (24, 48 and 96 h) with four detergents (CHAPS, SDS, SDC and Triton X-100) were used for decellularizing rat lungs for maintaining of three-dimensional lung architecture and ECM protein composition which have significant roles in differentiation and migration of stem cells. This comparative study determined that variable decellularization approaches can cause significantly different effects on decellularized lungs. RESULTS: Results showed that destruction was increased with increasing the detergent concentration. Single detergent showed a significant reduction in maintaining of three-dimensional of lung and ECM proteins (Collagen and Elastin). But, the best methods were mixed detergents of SDC and CHAPS in low concentration in 48 and 96 h decellularization. CONCLUSION: Decellularized lung tissue can be used in the laboratory to study various aspects of pulmonary biology and physiology and also, these results can be used in the continued improvement of engineered lung tissue. PMID:29362610

  16. Removal of detergents from proteins and peptides in a spin-column format.

    PubMed

    Antharavally, Babu S

    2012-08-01

    To enable downstream analysis, it is critical to remove unbound detergents from protein and peptide samples. This unit describes the use of a high-performance resin that offers exceptional detergent removal for proteins and peptides. The easy-to-use spin format significantly improves results over the standard drip column and batch methodologies, with >95% removal of 1% to 5% detergents, including SDS, sodium deoxycholate, CHAPS, Triton X-100, Triton X-114, NP-40, Brij-35, octyl glucoside, octyl thioglucoside, and lauryl maltoside, with high recovery of proteins and peptides. Detergent removal efficiency is evaluated using colorimetric methods and mass spectrometry (MS). BSA tryptic peptides have been successfully analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and matrix-assisted laser desorption/ionization (MALDI)-MS for identification of protein, following detergent removal using the resin. Advantages of this method include speed (less than 15 min), efficient detergent removal, and high recovery of proteins and peptides. © 2012 by John Wiley & Sons, Inc.

  17. Differential antigenic protein recovery from Taenia solium cyst tissues using several detergents.

    PubMed

    Navarrete-Perea, José; Orozco-Ramírez, Rodrigo; Moguel, Bárbara; Sciutto, Edda; Bobes, Raúl J; Laclette, Juan P

    2015-07-01

    Human and porcine cysticercosis is caused by the larval stage of the flatworm Taenia solium (Cestoda). The protein extracts of T. solium cysts are complex mixtures including cyst's and host proteins. Little is known about the influence of using different detergents in the efficiency of solubilization-extraction of these proteins, including relevant antigens. Here, we describe the use of CHAPS, ASB-14 and Triton X-100, alone or in combination in the extraction buffers, as a strategy to notably increase the recovery of proteins that are usually left aside in insoluble fractions of cysts. Using buffer with CHAPS alone, 315 protein spots were detected through 2D-PAGE. A total of 255 and 258 spots were detected using buffers with Triton X-100 or ASB-14, respectively. More protein spots were detected when detergents were combined, i.e., 2% CHAPS, 1% Triton X-100 and 1% ASB-14 allowed detection of up to 368 spots. Our results indicated that insoluble fractions of T. solium cysts were rich in antigens, including several glycoproteins that were sensitive to metaperiodate treatment. Host proteins, a common component in protein extracts of cysts, were present in larger amounts in soluble than insoluble fractions of cysts proteins. Finally, antigens present in the insoluble fraction were more appropriate as a source of antigens for diagnostic procedures. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Electrodynamics of Lipid Membrane Interactions in the Presence of Zwitterionic Buffers

    PubMed Central

    Koerner, Megan M.; Palacio, Luis A.; Wright, Johnnie W.; Schweitzer, Kelly S.; Ray, Bruce D.; Petrache, Horia I.

    2011-01-01

    Due to thermal motion and molecular polarizability, electrical interactions in biological systems have a dynamic character. Zwitterions are dipolar molecules that typically are highly polarizable and exhibit both a positive and a negative charge depending on the pH of the solution. We use multilamellar structures of common lipids to identify and quantify the effects of zwitterionic buffers that go beyond the control of pH. We use the fact that the repeat spacing of multilamellar lipid bilayers is a sensitive and accurate indicator of the force balance between membranes. We show that common buffers can in fact charge up neutral membranes. However, this electrostatic effect is not immediately recognized because of the concomitant modification of dispersion (van der Waals) forces. We show that although surface charging can be weak, electrostatic forces are significant even at large distances because of reduced ionic screening and reduced van der Waals attraction. The zwitterionic interactions that we identify are expected to be relevant for interfacial biological processes involving lipid bilayers, and for a wide range of biomaterials, including amino acids, detergents, and pharmaceutical drugs. An appreciation of zwitterionic electrodynamic character can lead to a better understanding of molecular interactions in biological systems and in soft materials in general. PMID:21767488

  19. Detergent effects on enzyme activity and solubilization of lipid bilayer membranes.

    PubMed

    Womack, M D; Kendall, D A; MacDonald, R C

    1983-09-07

    Over 50 detergents were tested to establish which would be most effective in releasing proteins from membrane-bounded compartments without denaturing them. Various concentrations each of detergent were tested for two activities: (1) solubilization of egg phospholipid liposomes as measured by reduction of turbidity and (2) effect of detergent concentration on the activities of soluble, hydrolytic enzymes. Those detergents most effective in solubilizing 0.2% lipid and least detrimental to enzymes were five pure, synthetic compounds recently introduced: CHAPS, CHAPSO, Zwittergents 310 and 312, and octylglucoside. Industrial detergents were generally much inferior, insofar as they solubilized membranes inefficiently and/or inactivated certain hydrolytic enzymes readily. The five detergents were characterized by (a) an unusually high critical micelle concentration and (b) a preference for forming mixed micelles with lipids instead of forming pure micelles, as indicated by an ability to solubilize lipid at concentrations of detergent significantly below the critical micelle concentration. This characteristic permits solubilization of high concentrations of membrane below the critical micelle concentration of the detergent so that protein denaturation is minimized. A generally applicable guideline that emerged from this study is that detergents should be used at approximately their critical micelle concentration which should not be exceeded by the concentration of membrane. Similar considerations should apply to the use of detergents in purifying and reconstituting intrinsic membrane proteins.

  20. A survey of detergents for the purification of stable, active human cystic fibrosis transmembrane conductance regulator (CFTR).

    PubMed

    Hildebrandt, Ellen; Zhang, Qinghai; Cant, Natasha; Ding, Haitao; Dai, Qun; Peng, Lingling; Fu, Yu; DeLucas, Lawrence J; Ford, Robert; Kappes, John C; Urbatsch, Ina L

    2014-11-01

    Structural knowledge of the cystic fibrosis transmembrane conductance regulator (CFTR) requires developing methods to purify and stabilize this aggregation-prone membrane protein above 1mg/ml. Starting with green fluorescent protein- and epitope-tagged human CFTR produced in mammalian cells known to properly fold and process CFTR, we devised a rapid tandem affinity purification scheme to minimize CFTR exposure to detergent in order to preserve its ATPase function. We compared a panel of detergents, including widely used detergents (maltosides, neopentyl glycols (MNG), C12E8, lysolipids, Chaps) and innovative detergents (branched alkylmaltosides, facial amphiphiles) for CFTR purification, function, monodispersity and stability. ATPase activity after reconstitution into proteoliposomes was 2-3 times higher when CFTR was purified using facial amphiphiles. ATPase activity was also demonstrated in purified CFTR samples without detergent removal using a novel lipid supplementation assay. By electron microscopy, negatively stained CFTR samples were monodisperse at low concentration, and size exclusion chromatography showed a predominance of monomer even after CFTR concentration above 1mg/ml. Rates of CFTR aggregation quantified in an electrophoretic mobility shift assay showed that detergents which best preserved reconstituted ATPase activity also supported the greatest stability, with CFTR monomer half-lives of 6-9days in MNG or Chaps, and 12-17days in facial amphiphile. Cryoelectron microscopy of concentrated CFTR in MNG or facial amphiphile confirmed mostly monomeric protein, producing low resolution reconstructions in conformity with similar proteins. These protocols can be used to generate samples of pure, functional, stable CFTR at concentrations amenable to biophysical characterization. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. A Survey of Detergents for the Purification of Stable, Active Human Cystic Fibrosis Transmembrane Conductance Regulator (CFTR)

    PubMed Central

    Hildebrandt, Ellen; Zhang, Qinghai; Cant, Natasha; Ding, Haitao; Dai, Qun; Peng, Lingling; Fu, Yu; DeLucas, Lawrence J.; Ford, Robert; Kappes, John C.; Urbatsch, Ina L.

    2014-01-01

    Structural knowledge of the cystic fibrosis transmembrane conductance regulator (CFTR) requires developing methods to purify and stabilize this aggregation-prone membrane protein above 1 mg/ml. Starting with green fluorescent protein- and epitope-tagged human CFTR produced in mammalian cells known to properly fold and process CFTR, we devised a rapid tandem affinity purification scheme to minimize CFTR exposure to detergent in order to preserve its ATPase function. We compared a panel of detergents, including widely used detergents (maltosides, neopentyl gycols (MNG), C12E8, lysolipids, Chaps) and innovative detergents (branched alkylmaltosides, facial amphiphiles) for CFTR purification, function, monodispersity and stability. ATPase activity after reconstitution into proteoliposomes was 2–3 times higher when CFTR was purified using facial amphiphiles. ATPase activity was also demonstrated in purified CFTR samples without detergent removal using a novel lipid supplementation assay. By electron microscopy, negatively stained CFTR samples were monodisperse at low concentration, and size exclusion chromatography showed a predominance of monomer even after CFTR concentration above 1 mg/ml. Rates of CFTR aggregation quantified in an electrophoretic mobility shift assay showed that detergents which best preserved reconstituted ATPase activity also supported the greatest stability, with CFTR monomer half-lives of 6–9 days in MNG or Chaps, and 12–17 days in facial amphiphile. Cryoelectron microscopy of concentrated CFTR in MNG or facial amphiphile confirmed mostly monomeric protein, producing low resolution reconstructions in conformity with similar proteins. These protocols can be used to generate samples of pure, functional, stable CFTR at concentrations amenable to biophysical characterization. PMID:25065669

  2. Size And Shape of Detergent Micelles Determined By Small-Angle X-Ray Scattering

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

    Lipfert, Jan; Columbus, Linda; Chu, Vincent B.

    2009-04-29

    We present a systematic analysis of the aggregation number and shape of micelles formed by nine detergents commonly used in the study of membrane proteins. Small-angle X-ray scattering measurements are reported for glucosides with 8 and 9 alkyl carbons (OG/NG), maltosides and phosphocholines with 10 and 12 alkyl carbons (DM/DDM and FC-10/FC-12), 1,2-dihexanoyl-sn-glycero-phosphocholine (DHPC), 1-palmitoyl-2-hydroxy-sn-glycero-3-[phospho-rac-(1-glycerol)] (LPPG), and 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS). The SAXS intensities are well described by two-component ellipsoid models, with a dense outer shell corresponding to the detergent head groups and a less electron dense hydrophobic core. These models provide an intermediate resolution view of micelle size and shape.more » In addition, we show that Guinier analysis of the forward scattering intensity can be used to obtain an independent and model-free measurement of the micelle aggregation number and radius of gyration. This approach has the advantage of being easily generalizable to protein-detergent complexes, where simple geometric models are inapplicable. Furthermore, we have discovered that the position of the second maximum in the scattering intensity provides a direct measurement of the characteristic head group-head group spacing across the micelle core. Our results for the micellar aggregation numbers and dimensions agree favorably with literature values as far as they are available. We de novo determine the shape of FC-10, FC-12, DM, LPPG, and CHAPS micelles and the aggregation numbers of FC-10 and OG to be ca. 50 and 250, respectively. Combined, these data provide a comprehensive view of the determinants of micelle formation and serve as a starting point to correlate detergent properties with detergent-protein interactions.« less

  3. Impact of Detergents on Membrane Protein Complex Isolation.

    PubMed

    Lee, Yu-Chen; Bååth, Jenny Arnling; Bastle, Ryan M; Bhattacharjee, Sonali; Cantoria, Mary Jo; Dornan, Mark; Gamero-Estevez, Enrique; Ford, Lenzie; Halova, Lenka; Kernan, Jennifer; Kürten, Charlotte; Li, Siran; Martinez, Jerahme; Sachan, Nalani; Sarr, Medoune; Shan, Xiwei; Subramanian, Nandhitha; Rivera, Keith; Pappin, Darryl; Lin, Sue-Hwa

    2018-01-05

    Detergents play an essential role during the isolation of membrane protein complexes. Inappropriate use of detergents may affect the native fold of the membrane proteins, their binding to antibodies, or their interaction with partner proteins. Here we used cadherin-11 (Cad11) as an example to examine the impact of detergents on membrane protein complex isolation. We found that mAb 1A5 could immunoprecipitate Cad11 when membranes were solubilized by dodecyl maltoside (DDM) but not by octylglucoside, suggesting that octylglucoside interferes with Cad11-mAb 1A5 interaction. Furthermore, we compared the effects of Brij-35, Triton X-100, cholate, CHAPSO, Zwittergent 3-12, Deoxy BIG CHAP, and digitonin on Cad11 solubilization and immunoprecipitation. We found that all detergents except Brij-35 could solubilize Cad11 from the membrane. Upon immunoprecipitation, we found that β-catenin, a known cadherin-interacting protein, was present in Cad11 immune complex among the detergents tested except Brij-35. However, the association of p120 catenin with Cad11 varied depending on the detergents used. Using isobaric tag for relative and absolute quantitation (iTRAQ) to determine the relative levels of proteins in Cad11 immune complexes, we found that DDM and Triton X-100 were more efficient than cholate in solubilization and immunoprecipitation of Cad11 and resulted in the identification of both canonical and new candidate Cad11-interacting proteins.

  4. Effect of Detergents on the Thermal Behavior of Elastin-like Polypeptides

    PubMed Central

    Thapa, Arjun; Han, Wei; Simons, Robin H.; Chilkoti, Ashutosh; Chi, Eva Y.; López, Gabriel P.

    2012-01-01

    Elastin-like polypeptide (ELP) fusions have been designed to allow large scale, non-chromatographic purification of many soluble proteins using the inverse transition cycling (ITC) method; however, the sensitivity of the aqueous lower critical solubility phase transition temperature (Tt) of ELPs to the addition of cosolutes, including detergents, may be a potential hindrance in purification of proteins with surface hydrophobicity in such a manner. To identify detergents that are known to solubilize such proteins (e.g., membrane proteins) and that have little effect on the Tt of the ELP, we screened a number of detergents with respect to their effects on the Tt and secondary structures of a model ELP (denoted here as ELP180). We found that mild detergents (e.g., DDM, Triton-X100, and CHAPS) do not alter the phase transition behavior or structure (as probed by circular dichroism) of ELP180. This result is in contrast to previous studies that showed a strong effect of other detergents (e.g., SDS) on the Ttof ELPs. Our results clearly indicate that mild detergents do not preclude ITC-based separation of ELPs, and thus that ELP fusions may prove to be useful in the purification of detergent-solubilized recombinant hydrophobic proteins, including membrane proteins, which are otherwise notoriously difficult to extract and purify by conventional separation methods (e.g., chromatography). PMID:23097230

  5. Visualization of Detergent Solubilization of Membranes: Implications for the Isolation of Rafts

    PubMed Central

    Garner, Ashley E.; Smith, D. Alastair; Hooper, Nigel M.

    2008-01-01

    Although different detergents can give rise to detergent-resistant membranes of different composition, it is unclear whether this represents domain heterogeneity in the original membrane. We compared the mechanism of action of five detergents on supported lipid bilayers composed of equimolar sphingomyelin, cholesterol, and dioleoylphosphatidylcholine imaged by atomic force microscopy, and on raft and nonraft marker proteins in live cells imaged by confocal microscopy. There was a marked correlation between the detergent solubilization of the cell membrane and that of the supported lipid bilayers. In both systems Triton X-100 and CHAPS (3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate) distinguished between the nonraft liquid-disordered (ld) and raft liquid ordered (lo) lipid phases by selectively solubilizing the ld phase. A higher concentration of Lubrol was required, and not all the ld phase was solubilized. The solubilization by Brij 96 occurred by a two-stage mechanism that initially resulted in the solubilization of some ld phase and then progressed to the solubilization of both ld and lo phases simultaneously. Octyl glucoside simultaneously solubilized both lo and ld phases. These data show that the mechanism of membrane solubilization is unique to an individual detergent. Our observations have significant implications for using different detergents to isolate membrane rafts from biological systems. PMID:17933878

  6. Visualization of detergent solubilization of membranes: implications for the isolation of rafts.

    PubMed

    Garner, Ashley E; Smith, D Alastair; Hooper, Nigel M

    2008-02-15

    Although different detergents can give rise to detergent-resistant membranes of different composition, it is unclear whether this represents domain heterogeneity in the original membrane. We compared the mechanism of action of five detergents on supported lipid bilayers composed of equimolar sphingomyelin, cholesterol, and dioleoylphosphatidylcholine imaged by atomic force microscopy, and on raft and nonraft marker proteins in live cells imaged by confocal microscopy. There was a marked correlation between the detergent solubilization of the cell membrane and that of the supported lipid bilayers. In both systems Triton X-100 and CHAPS (3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate) distinguished between the nonraft liquid-disordered (l(d)) and raft liquid ordered (l(o)) lipid phases by selectively solubilizing the l(d) phase. A higher concentration of Lubrol was required, and not all the l(d) phase was solubilized. The solubilization by Brij 96 occurred by a two-stage mechanism that initially resulted in the solubilization of some l(d) phase and then progressed to the solubilization of both l(d) and l(o) phases simultaneously. Octyl glucoside simultaneously solubilized both l(o) and l(d) phases. These data show that the mechanism of membrane solubilization is unique to an individual detergent. Our observations have significant implications for using different detergents to isolate membrane rafts from biological systems.

  7. Effect of surfactants and natural detergents on phosphatidylcholine synthesis in photoreceptor membranes.

    PubMed

    Roque, M E; Castagnet, P I; Giusto, N M

    2001-07-01

    The synthesis of phosphatidylcholine (PC) in rod outer segments (ROS) catalysed by lysophosphatidylcholine acyltransferase and phosphatidylethanolamine N-methyltransferase (PE N-MTase) was studied and the effects of natural (FA and lysophospholipids) and synthetic (Triton X-100, deoxycholate and CHAPS) surfactants was evaluated. In all experimental conditions used, incorporation of labelled oleate into lysophosphatidylcholine (lysoPC) was at least 40 times greater than oleate incorporation into any other lysophospholipid. Acylation of lysoPC was slightly affected by Triton X-100 and was totally inhibited in the presence of 10 mM sodium deoxycholate (NaDOC) or CHAPS. Below their critical micelle concentration (cmc) Triton X-100 and NaDOC stimulated acylation of all ROS lysophospholipids analysed. The activity of PE N-MTase was stimulated at detergent concentrations below the cmc and inhibited at concentrations above the cmc for all three detergents tested. The effect of FA with differing degree of unsaturation on PC synthesis was evaluated. Oleic acid (10 microM) inhibited methyl group incorporation into total PC, whereas from 100 microM onward, the methylating activity increased with preferential synthesis of PC. Docosahexaenoic acid, in turn, inhibited PE N-MTase activity at every concentration tested. These results suggest that PC synthesis in ROS membranes is modified by bioregulators and surfactants altering the physico-chemical state of the membrane.

  8. Conjugated polymer zwitterions and solar cells comprising conjugated polymer zwitterions

    DOEpatents

    Emrick, Todd; Russell, Thomas; Page, Zachariah; Liu, Yao

    2018-06-05

    A conjugated polymer zwitterion includes repeating units having structure (I), (II), or a combination thereof ##STR00001## wherein Ar is independently at each occurrence a divalent substituted or unsubstituted C3-30 arylene or heteroarylene group; L is independently at each occurrence a divalent C1-16 alkylene group, C6-30arylene or heteroarylene group, or alkylene oxide group; and R1 is independently at each occurrence a zwitterion. A polymer solar cell including the conjugated polymer zwitterion is also disclosed.

  9. Structure and Dynamics of Dinucleosomes Assessed by Atomic Force Microscopy

    DOE PAGES

    Filenko, Nina A.; Palets, Dmytro B.; Lyubchenko, Yuri L.

    2012-01-01

    Dynamics of nucleosomes and their interactions are important for understanding the mechanism of chromatin assembly. Internucleosomal interaction is required for the formation of higher-order chromatin structures. Although H1 histone is critically involved in the process of chromatin assembly, direct internucleosomal interactions contribute to this process as well. To characterize the interactions of nucleosomes within the nucleosome array, we designed a dinucleosome and performed direct AFM imaging. The analysis of the AFM data showed dinucleosomes are very dynamic systems, enabling the nucleosomes to move in a broad range along the DNA template. Di-nucleosomes in close proximity were observed, but their populationmore » was low. The use of the zwitterionic detergent, CHAPS, increased the dynamic range of the di-nucleosome, facilitating the formation of tight di-nucleosomes. The role of CHAPS and similar natural products in chromatin structure and dynamics is also discussed.« less

  10. Additive and Synergistic Membrane Permeabilization by Antimicrobial (Lipo)Peptides and Detergents

    PubMed Central

    Patel, Hiren; Huynh, Quang; Bärlehner, Dominik; Heerklotz, Heiko

    2014-01-01

    Certain antibiotic peptides are thought to permeabilize membranes of pathogens by effects that are also observed for simple detergents, such as membrane thinning and disordering, asymmetric bilayer expansion, toroidal pore formation, and micellization. Here we test the hypothesis that such peptides act additively with detergents when applied in parallel. Additivity is defined analogously to a fractional inhibitory concentration index of unity, and the extent and mechanism of leakage is measured by the fluorescence lifetime-based vesicle leakage assay using calcein-loaded vesicles. Good additivity was found for the concerted action of magainin 2, the fungicidal lipopeptide class of surfactins from Bacillus subtilis QST713, and the detergent octyl glucoside, respectively, with the detergent C12EO8. Synergistic or superadditive action was observed for fengycins from B. subtilis, as well as the detergent CHAPS, when combined with C12EO8. The results illustrate two mechanisms of synergistic action: First, maximal leakage requires an optimum degree of heterogeneity in the system that may be achieved by mixing a graded with an all-or-none permeabilizer. (The optimal perturbation should be focused to certain defect structures, yet not to the extent that some vesicles are not affected at all.) Second, a cosurfactant may enhance the bioavailability of a poorly soluble peptide. The results are important for understanding the concerted action of membrane-permeabilizing compounds in biology as well as for optimizing formulations of such antimicrobials for medical applications or crop protection. PMID:24853740

  11. Micro-scale NMR Screening of New Detergents for Membrane Protein Structural Biology

    PubMed Central

    Zhang, Qinghai; Horst, Reto; Geralt, Michael; Ma, Xingquan; Hong, Wen-Xu; Finn, M. G.; Stevens, Raymond C.; Wüthrich, Kurt

    2008-01-01

    The rate limiting step in biophysical characterization of membrane proteins is often the availability of suitable amounts of protein material. It was therefore of interest to demonstrate that micro-coil nuclear magnetic resonance (NMR) technology can be used to screen microscale quantities of membrane proteins for proper folding in samples destined for structural studies. Micoscale NMR was then used to screen a series of newly designed zwitterionic phosphocholine detergents for their ability to reconstitute membrane proteins, using the previously well characterized β-barrel E.coli outer membrane protein OmpX as a test case. Fold screening was thus achieved with μg-amounts of uniformly 2H,15N-labeld OmpX and affordable amounts of the detergents, and prescreening with SDS-gel electrophoresis ensured efficient selection of the targets for NMR studies. A systematic approach to optimize the phosphocholine motif for membrane protein refolding led to the identification of two new detergents, 138-Fos and 179-Fos, that yield 2D [15N,1H]-TROSY correlation NMR spectra of natively folded reconstituted OmpX. PMID:18479092

  12. Production of functional bacteriorhodopsin by an Escherichia coli cell-free protein synthesis system supplemented with steroid detergent and lipid.

    PubMed

    Shimono, Kazumi; Goto, Mie; Kikukawa, Takashi; Miyauchi, Seiji; Shirouzu, Mikako; Kamo, Naoki; Yokoyama, Shigeyuki

    2009-10-01

    Cell-free expression has become a highly promising tool for the efficient production of membrane proteins. In this study, we used a dialysis-based Escherichia coli cell-free system for the production of a membrane protein actively integrated into liposomes. The membrane protein was the light-driven proton pump bacteriorhodopsin, consisting of seven transmembrane alpha-helices. The cell-free expression system in the dialysis mode was supplemented with a combination of a detergent and a natural lipid, phosphatidylcholine from egg yolk, in only the reaction mixture. By examining a variety of detergents, we found that the combination of a steroid detergent (digitonin, cholate, or CHAPS) and egg phosphatidylcholine yielded a large amount (0.3-0.7 mg/mL reaction mixture) of the fully functional bacteriorhodopsin. We also analyzed the process of functional expression in our system. The synthesized polypeptide was well protected from aggregation by the detergent-lipid mixed micelles and/or lipid disks, and was integrated into liposomes upon detergent removal by dialysis. This approach might be useful for the high yield production of functional membrane proteins.

  13. Integrated antimicrobial and nonfouling zwitterionic polymers.

    PubMed

    Mi, Luo; Jiang, Shaoyi

    2014-02-10

    Zwitterionic polymers are generally viewed as a new class of nonfouling materials. Unlike their poly(ethylene glycol) (PEG) counterparts, zwitterionic polymers have a broader chemical diversity and greater freedom for molecular design. In this Minireview, we highlight recent microbiological applications of zwitterionic polymers and their derivatives, with an emphasis on several unique molecular strategies to integrate antimicrobial and nonfouling properties. We will also discuss our insights into the bacterial nonfouling performance of zwitterionic polymers and one example of engineering zwitterionic polymer derivatives for antimicrobial wound-dressing applications. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Solubilization of adenylyl cyclase from human myometrium in a alphas-coupled form.

    PubMed

    Bajo, Ana M; Prieto, Juan C; Valenzuela, Pedro; Martinez, Pilar; Guijarro, Luis G

    2003-08-01

    Adenylyl cyclase (AC) was extracted from human myometrium with either non-ionic (Lubrol-PX or Triton X-100) or zwitterionic (3-[3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, CHAPS) detergents. The soluble enzyme was stimulated by forskolin, a hydrophobic activator, in the presence of Mg2+ indicating that the catalytic subunit had not been damaged after solubilization. The enzyme was also activated by 5'-guanylyl imidodiphosphate (Gpp(NH)p) showing that the catalytic unit was not separated from stimulatory guanine nucleotide binding protein (Gs) during the extraction. Both activators showed different effects on the stimulatory efficacy and potency of AC activity solobulized with detergents. Gel filtration of Lubrol-PX and CHAPS extracts over a Sepharose CL-2B column partially resolved AC and its complexes. The chromatographic profile for Lubrol-solubilized AC presented a main peak of about 200 kDa whereas CHAPS-solubilized AC showed a dominant peak of about 1100 kDa. The heterodisperse peaks obtained revealed that the catalytic AC subunit was not separated from Gs proteins after gel filtration, and that AC could be associated with other cellular proteins. When Lubrol extract was submitted to anionic-exchange chromatography, the enzyme was purified about 7.5 fold (enzymatic activity of 48.1 pmol/min/mg of protein). The catalytic subunit was co-eluted with both AC-activating proteins Galphas large (52.2 kDa) and Galphas small (48.7 kDa). This is the first demonstration of the stable physical association of AC with both alphas subunits of G proteins in human myometrium.

  15. The impact of detergents on the tissue decellularization process: a ToF-SIMS study

    PubMed Central

    White, Lisa J; Taylor, Adam J; Faulk, Denver M; Keane, Tim J; Saldin, Lindsey T; Reing, Janet E; Swinehart, Ilea T; Turner, Neill J; Ratner, Buddy D

    2017-01-01

    Biologic scaffolds are derived from mammalian tissues, which must be decellularized to remove cellular antigens that would otherwise incite an adverse immune response. Although widely used clinically, the optimum balance between cell removal and the disruption of matrix architecture and surface ligand landscape remains a considerable challenge. Here we describe the use of time of flight secondary ion mass spectroscopy (ToF-SIMS) to provide sensitive, molecular specific, localized analysis of detergent decellularized biologic scaffolds. We detected residual detergent fragments, specifically from Triton X-100, sodium deoxycholate and sodium dodecyl sulphate (SDS) in decellularized scaffolds; increased SDS concentrations from 0.1% to 1.0% increased both the intensity of SDS fragments and adverse cell outcomes. We also identified cellular remnants, by detecting phosphate and phosphocholine ions in PAA and CHAPS decellularized scaffolds. The present study demonstrates ToF-SIMS is not only a powerful tool for characterization of biologic scaffold surface molecular functionality, but also enables sensitive assessment of decellularization efficacy. PMID:27993639

  16. Zwitterionic materials for antifouling membrane surface construction.

    PubMed

    He, Mingrui; Gao, Kang; Zhou, Linjie; Jiao, Zhiwei; Wu, Mengyuan; Cao, Jialin; You, Xinda; Cai, Ziyi; Su, Yanlei; Jiang, Zhongyi

    2016-08-01

    Membrane separation processes are often perplexed by severe and ubiquitous membrane fouling. Zwitterionic materials, keeping electric neutrality with equivalent positive and negative charged groups, are well known for their superior antifouling properties and have been broadly utilized to construct antifouling surfaces for medical devices, biosensors and marine coatings applications. In recent years, zwitterionic materials have been more and more frequently utilized for constructing antifouling membrane surfaces. In this review, the antifouling mechanisms of zwitterionic materials as well as their biomimetic prototypes in cell membranes will be discussed, followed by the survey of common approaches to incorporate zwitterionic materials onto membrane surfaces including surface grafting, surface segregation, biomimetic adhesion, surface coating and so on. The potential applications of these antifouling membranes are also embedded. Finally, we will present a brief perspective on the future development of zwitterionic materials modified antifouling membranes. Membrane fouling is a severe problem hampering the application of membrane separation technology. The properties of membrane surfaces play a critical role in membrane fouling and antifouling behavior/performance. Antifouling membrane surface construction has evolved as a hot research issue for the development of membrane processes. Zwitterionic modification of membrane surfaces has been recognized as an effective strategy to resist membrane fouling. This review summarizes the antifouling mechanisms of zwitterionic materials inspired by cell membranes as well as the popular approaches to incorporate them onto membrane surfaces. It can help form a comprehensive knowledge about the principles and methods of modifying membrane surfaces with zwitterionic materials. Finally, we propose the possible future research directions of zwitterionic materials modified antifouling membranes. Copyright © 2016 Acta Materialia Inc

  17. Cell Nucleus-Targeting Zwitterionic Carbon Dots.

    PubMed

    Jung, Yun Kyung; Shin, Eeseul; Kim, Byeong-Su

    2015-12-22

    An innovative nucleus-targeting zwitterionic carbon dot (CD) vehicle has been developed for anticancer drug delivery and optical monitoring. The zwitterionic functional groups of the CDs introduced by a simple one-step synthesis using β-alanine as a passivating and zwitterionic ligand allow cytoplasmic uptake and subsequent nuclear translocation of the CDs. Moreover, multicolor fluorescence improves the accuracy of the CDs as an optical code. The CD-based drug delivery system constructed by non-covalent grafting of doxorubicin, exhibits superior antitumor efficacy owing to enhanced nuclear delivery in vitro and tumor accumulation in vivo, resulting in highly effective tumor growth inhibition. Since the zwitterionic CDs are highly biocompatible and effectively translocated into the nucleus, it provides a compelling solution to a multifunctional nanoparticle for substantially enhanced nuclear uptake of drugs and optical monitoring of translocation.

  18. The impact of detergents on the tissue decellularization process: A ToF-SIMS study.

    PubMed

    White, Lisa J; Taylor, Adam J; Faulk, Denver M; Keane, Timothy J; Saldin, Lindsey T; Reing, Janet E; Swinehart, Ilea T; Turner, Neill J; Ratner, Buddy D; Badylak, Stephen F

    2017-03-01

    Biologic scaffolds are derived from mammalian tissues, which must be decellularized to remove cellular antigens that would otherwise incite an adverse immune response. Although widely used clinically, the optimum balance between cell removal and the disruption of matrix architecture and surface ligand landscape remains a considerable challenge. Here we describe the use of time of flight secondary ion mass spectroscopy (ToF-SIMS) to provide sensitive, molecular specific, localized analysis of detergent decellularized biologic scaffolds. We detected residual detergent fragments, specifically from Triton X-100, sodium deoxycholate and sodium dodecyl sulphate (SDS) in decellularized scaffolds; increased SDS concentrations from 0.1% to 1.0% increased both the intensity of SDS fragments and adverse cell outcomes. We also identified cellular remnants, by detecting phosphate and phosphocholine ions in PAA and CHAPS decellularized scaffolds. The present study demonstrates ToF-SIMS is not only a powerful tool for characterization of biologic scaffold surface molecular functionality, but also enables sensitive assessment of decellularization efficacy. We report here on the use of a highly sensitive analytical technique, time of flight secondary ion mass spectroscopy (ToF-SIMS) to characterize detergent decellularized scaffolds. ToF-SIMS detected cellular remnants and residual detergent fragments; increased intensity of the detergent fragments correlated with adverse cell matrix interactions. This study demonstrates the importance of maintaining a balance between cell removal and detergent disruption of matrix architecture and matrix surface ligand landscape. This study also demonstrates the power of ToF-SIMS for the characterization of decellularized scaffolds and capability for assessment of decellularization efficacy. Future use of biologic scaffolds in clinical tissue reconstruction will benefit from the fundamental results described in this work. Copyright © 2016 Acta

  19. Cell Nucleus-Targeting Zwitterionic Carbon Dots

    PubMed Central

    Jung, Yun Kyung; Shin, Eeseul; Kim, Byeong-Su

    2015-01-01

    An innovative nucleus-targeting zwitterionic carbon dot (CD) vehicle has been developed for anticancer drug delivery and optical monitoring. The zwitterionic functional groups of the CDs introduced by a simple one-step synthesis using β-alanine as a passivating and zwitterionic ligand allow cytoplasmic uptake and subsequent nuclear translocation of the CDs. Moreover, multicolor fluorescence improves the accuracy of the CDs as an optical code. The CD-based drug delivery system constructed by non-covalent grafting of doxorubicin, exhibits superior antitumor efficacy owing to enhanced nuclear delivery in vitro and tumor accumulation in vivo, resulting in highly effective tumor growth inhibition. Since the zwitterionic CDs are highly biocompatible and effectively translocated into the nucleus, it provides a compelling solution to a multifunctional nanoparticle for substantially enhanced nuclear uptake of drugs and optical monitoring of translocation. PMID:26689549

  20. Detergent Isolation Stabilizes and Activates the Shigella Type III Secretion System Translocator Protein IpaC.

    PubMed

    Bernard, Abram R; Duarte, Shari M; Kumar, Prashant; Dickenson, Nicholas E

    2016-07-01

    Shigella rely on a type III secretion system as the primary virulence factor for invasion and colonization of human hosts. Although there are an estimated 90 million Shigella infections, annually responsible for more than 100,000 deaths worldwide, challenges isolating and stabilizing many type III secretion system proteins have prevented a full understanding of the Shigella invasion mechanism and additionally slowed progress toward a much needed Shigella vaccine. Here, we show that the non-denaturing zwitterionic detergent N, N-dimethyldodecylamine N-oxide (LDAO) and non-ionic detergent n-octyl-oligo-oxyethylene efficiently isolated the hydrophobic Shigella translocator protein IpaC from the co-purified IpaC/IpgC chaperone-bound complex. Both detergents resulted in monomeric IpaC that exhibits strong membrane binding and lysis characteristics while the chaperone-bound complex does not, suggesting that the stabilizing detergents provide a means of following IpaC "activation" in vitro. Additionally, biophysical characterization found that LDAO provides significant thermal and temporal stability to IpaC, protecting it for several days at room temperature and brief exposure to temperatures reaching 90°C. In summary, this work identified and characterized conditions that provide stable, membrane active IpaC, providing insight into key interactions with membranes and laying a strong foundation for future vaccine formulation studies taking advantage of the native immunogenicity of IpaC and the stability provided by LDAO. Copyright © 2016 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

  1. Solubilization and characterization of haloperidol-sensitive (+)-( sup 3 H)SKF-10,047 binding sites (sigma sites) from rat liver membranes

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

    McCann, D.J.; Su, T.P.

    1991-05-01

    The zwitterionic detergent 3-((3-cholamidopropyl)dimethylamino)-1-propanesulfonate (CHAPS) produced optimal solubilization of (+)-({sup 3}H)SKF-10,047 binding sites from rat liver membranes at a concentration of 0.2%, well below the critical micellular concentration of the detergent. The pharmacological selectivity of the liver (+)-({sup 3}H)SKF-10,047 binding sites corresponds to that of sigma sites from rat and guinea pig brain. When the affinities of 18 different drugs at (+)-({sup 3}H)SKF-10,047 binding sites in membranes and solubilized preparations were compared, a correlation coefficient of 0.99 and a slope of 1.03 were obtained, indicating that the pharmacological selectivity of rat liver sigma sites is retained after solubilization. In addition,more » the binding of 20 nM ({sup 3}H)progesterone to solubilized rat liver preparations was found to exhibit a pharmacological selectivity appropriate for sigma sites. A stimulatory effect of phenytoin on (+)-({sup 3}H)SKF-10,047 binding to sigma sites persisted after solubilization. When the solubilized preparation was subjected to molecular sizing chromatography, a single peak exhibiting specific (+)-({sup 3}H)SKF-10,047 binding was obtained. The binding activity of this peak was stimulated symmetrically when assays were performed in the presence of 300 microM phenytoin. The molecular weight of the CHAPS-solubilized sigma site complex was estimated to be 450,000 daltons. After solubilization with CHAPS, rat liver sigma sites were enriched to 12 pmol/mg of protein. The present results demonstrate a successful solubilization of sigma sites from rat liver membranes and provide direct evidence that the gonadal steroid progesterone binds to sigma sites. The results also suggest that the anticonvulsant phenytoin binds to an associated allosteric site on the sigma site complex.« less

  2. Correlating antimicrobial activity and model membrane leakage induced by nylon-3 polymers and detergents

    PubMed Central

    Hovakeemian, Sara G.; Liu, Runhui; Gellman, Samuel H.; Heerklotz, Heiko

    2015-01-01

    Most antimicrobial peptides act upon target microorganisms by permeabilizing their membranes. The mode of action is often assessed by vesicle leakage experiments that use model membranes, with the assumption that biological activity arises from permeabilization of the lipid bilayer. The current work aims to extend the interpretation of vesicle leakage results and examine the correlation between vesicle leakage and antimicrobial activity. To this end, we used a lifetime-based leakage assay with calcein-loaded vesicles to study the membrane permeabilizing properties of a novel antifungal polymer poly-NM, two of its analogs, and a series of detergents. In conjunction, the biological activities of these compounds against Candida albicans were assessed and correlated with data from vesicle leakage. Poly-NM induces all-or-none leakage in polar yeast lipid vesicles at the polymer’s MIC, 3 μg/mL. At this and higher concentrations, complete leakage after an initial lag time was observed. Concerted activity tests imply that this polymer acts independently of the detergent octyl glucoside (OG) for both vesicle leakage and activity against C. albicans spheroplasts. In addition, Poly-NM was found to have negligible activity against zwitterionic vesicles and red blood cells. Our results provide a consistent, detailed picture of the mode of action of Poly-NM: this polymer induces membrane leakage by electrostatic lipid clustering. In contrast, Poly-MM:CO, a nylon-3 polymer comprised of both cationic and hydrophobic segments, seems to act by a different mechanism that involves membrane asymmetry stress. Vesicle leakage for this polymer is transient (limited to <100%) and graded, non-specific among zwitterionic and polar yeast lipid vesicles, additive with detergent action, and correlates poorly with biological activity. Based on these results, we conclude that comprehensive leakage experiments can provide a detailed description of the mode of action of membrane permeabilizing

  3. Correlating antimicrobial activity and model membrane leakage induced by nylon-3 polymers and detergents.

    PubMed

    Hovakeemian, Sara G; Liu, Runhui; Gellman, Samuel H; Heerklotz, Heiko

    2015-09-14

    Most antimicrobial peptides act upon target microorganisms by permeabilizing their membranes. The mode of action is often assessed by vesicle leakage experiments that use model membranes, with the assumption that biological activity correlates with the permeabilization of the lipid bilayer. The current work aims to extend the interpretation of vesicle leakage results and examine the correlation between vesicle leakage and antimicrobial activity. To this end, we used a lifetime-based leakage assay with calcein-loaded vesicles to study the membrane permeabilizing properties of a novel antifungal polymer poly-NM, two of its analogs, and a series of detergents. In conjunction, the biological activities of these compounds against Candida albicans were assessed and correlated with data from vesicle leakage. Poly-NM induces all-or-none leakage in polar yeast lipid vesicles at the polymer's MIC, 3 μg mL(-1). At this and higher concentrations, complete leakage after an initial lag time was observed. Concerted activity tests imply that this polymer acts independently of the detergent octyl glucoside (OG) for both vesicle leakage and activity against C. albicans spheroplasts. In addition, poly-NM was found to have negligible activity against zwitterionic vesicles and red blood cells. Our results provide a consistent, detailed picture of the mode of action of poly-NM: this polymer induces membrane leakage by electrostatic lipid clustering. In contrast, poly-MM:CO, a nylon-3 polymer comprised of both cationic and hydrophobic segments, seems to act by a different mechanism that involves membrane asymmetry stress. Vesicle leakage for this polymer is transient (limited to <100%) and graded, non-specific among zwitterionic and polar yeast lipid vesicles, additive with detergent action, and correlates poorly with biological activity. Based on these results, we conclude that comprehensive leakage experiments can provide a detailed description of the mode of action of membrane

  4. Total Protein Extraction for Metaproteomics Analysis of Methane Producing Biofilm: The Effects of Detergents

    PubMed Central

    Huang, Hung-Jen; Chen, Wei-Yu; Wu, Jer-Horng

    2014-01-01

    Protein recovery is crucial for shotgun metaproteomics to study the in situ functionality of microbial populations from complex biofilms but still poorly addressed by far. To fill this knowledge gap, we systematically evaluated the sample preparation with extraction buffers comprising four detergents for the metaproteomics analysis of a terephthalate-degrading methanogenic biofilm using an on-line two-dimensional liquid chromatography tandem mass spectrometry (2D-LC-MS/MS) system. Totally, 1018 non-repeated proteins were identified with the four treatments. On the whole, each treatment could recover the biofilm proteins with specific distributions of molecular weight, hydrophobicity, and isoelectric point. The extraction buffers containing zwitterionic and anionic detergents were found to harvest the proteins with better efficiency and quality, allowing identification up to 76.2% of total identified proteins with the LC-MS/MS analysis. According to the annotation with a relevant metagenomic database, we further observed different taxonomic profiles of bacterial and archaeal members and discriminable patterns of the functional expression among the extraction buffers used. Overall, the finding of the present study provides first insight to the effect of the detergents on the characteristics of extractable proteins from biofilm and the developed protocol combined with nano 2D-LC/MS/MS analysis can improve the metaproteomics studies on microbial functionality of biofilms in the wastewater treatment systems. PMID:24914765

  5. Biochemical and physical characterisation of urinary nanovesicles following CHAPS treatment.

    PubMed

    Musante, Luca; Saraswat, Mayank; Duriez, Elodie; Byrne, Barry; Ravidà, Alessandra; Domon, Bruno; Holthofer, Harry

    2012-01-01

    Urinary exosomes represent a precious source of potential biomarkers for disease biology. Currently, the methods for vesicle isolation are severely restricted by the tendency of vesicle entrapment, e.g. by the abundant Tamm-Horsfall protein (THP) polymers. Treatment by reducing agents such as dithiothreitol (DTT) releases entrapped vesicles, thus increasing the final yield. However, this harsh treatment can cause remodelling of all those proteins which feature extra-vesicular domains stabilized by internal disulfide bridges and have detrimental effects on their biological activity. In order to optimize exosomal yield, we explore two vesicle treatment protocols - dithiothreitol (DTT) and 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic (CHAPS) - applied to the differential centrifugation protocol for exosomal vesicle isolation. The results show that CHAPS treatment does not affect vesicle morphology or exosomal marker distribution, thus eliminating most of THP interference. Moreover, the recovery and preservation of catalytic activity of two trans-membrane proteases, dipeptidyl peptidase IV and nephrilysin, was examined and found to be clearly superior after CHAPS treatment compared to DTT. Finally, proteomic profiling by mass spectrometry (MS) revealed that 76.2% of proteins recovered by CHAPS are common to those seen for DTT treatment, which illustrates underlining similarities between the two approaches. In conclusion, we provide a major improvement to currently-utilized urinary vesicle isolation strategies to allow recovery of urinary vesicles without the deleterious interference of abundant urinary proteins, while preserving typical protein folding and, consequently, the precious biological activity of urinary proteins which serve as valuable biomarkers.

  6. CHAP III- CHARRING ABLATOR PROGRAM FOR ADVANCED INVESTIGATION OF THERMAL PROTECTION SYSTEMS FOR ENTRY

    NASA Technical Reports Server (NTRS)

    Stroud, C. W.

    1994-01-01

    The transient response of a thermal protection material to heat applied to the surface can be calculated using the CHAP III computer program. CHAP III can be used to analyze pyrolysis gas chemical kinetics in detail and examine pyrolysis reactions-indepth. The analysis includes the deposition of solid products produced by chemical reactions in the gas phase. CHAP III uses a modelling technique which can approximate a wide range of ablation problems. The energy equation used in CHAP III incorporates pyrolysis (both solid and gas reactions), convection, conduction, storage, work, kinetic energy, and viscous dissipation. The chemically reacting components of the solid are allowed to vary as a function of position and time. CHAP III employs a finite difference method to approximate the energy equations. Input values include specific heat, thermal conductivity, thermocouple locations, enthalpy, heating rates, and a description of the chemical reactions expected. The output tabulates the temperature at locations throughout the ablator, gas flow within the solid, density of the solid, weight of pyrolysis gases, and rate of carbon deposition. A sample case is included, which analyzes an ablator material containing several pyrolysis reactions subjected to an environment typical of entry at lunar return velocity. CHAP III is written in FORTRAN IV for batch execution and has been implemented on a CDC CYBER 170 series computer operating under NOS with a central memory requirement of approximately 102K (octal) of 60 bit words. This program was developed in 1985.

  7. User's manual for the Composite HTGR Analysis Program (CHAP-1)

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

    Gilbert, J.S.; Secker, P.A. Jr.; Vigil, J.C.

    1977-03-01

    CHAP-1 is the first release version of an HTGR overall plant simulation program with both steady-state and transient solution capabilities. It consists of a model-independent systems analysis program and a collection of linked modules, each representing one or more components of the HTGR plant. Detailed instructions on the operation of the code and detailed descriptions of the HTGR model are provided. Information is also provided to allow the user to easily incorporate additional component modules, to modify or replace existing modules, or to incorporate a completely new simulation model into the CHAP systems analysis framework.

  8. Experimenting with Detergents

    ERIC Educational Resources Information Center

    Mitchell, Gail; Phillips, Donald B.

    1977-01-01

    Lists materials and procedures for experimenting with detergents. Included are methods for determination of the densities of dry detergents, ph values of detergent solutions, and a discussion of the ability of detergents to remove iodine stains from cloth. (CS)

  9. Stabilization of Functional Recombinant Cannabinoid Receptor CB2 in Detergent Micelles and Lipid Bilayers

    PubMed Central

    Vukoti, Krishna; Kimura, Tomohiro; Macke, Laura; Gawrisch, Klaus; Yeliseev, Alexei

    2012-01-01

    Elucidation of the molecular mechanisms of activation of G protein-coupled receptors (GPCRs) is among the most challenging tasks for modern membrane biology. For studies by high resolution analytical methods, these integral membrane receptors have to be expressed in large quantities, solubilized from cell membranes and purified in detergent micelles, which may result in a severe destabilization and a loss of function. Here, we report insights into differential effects of detergents, lipids and cannabinoid ligands on stability of the recombinant cannabinoid receptor CB2, and provide guidelines for preparation and handling of the fully functional receptor suitable for a wide array of downstream applications. While we previously described the expression in Escherichia coli, purification and liposome-reconstitution of multi-milligram quantities of CB2, here we report an efficient stabilization of the recombinant receptor in micelles - crucial for functional and structural characterization. The effects of detergents, lipids and specific ligands on structural stability of CB2 were assessed by studying activation of G proteins by the purified receptor reconstituted into liposomes. Functional structure of the ligand binding pocket of the receptor was confirmed by binding of 2H-labeled ligand measured by solid-state NMR. We demonstrate that a concerted action of an anionic cholesterol derivative, cholesteryl hemisuccinate (CHS) and high affinity cannabinoid ligands CP-55,940 or SR-144,528 are required for efficient stabilization of the functional fold of CB2 in dodecyl maltoside (DDM)/CHAPS detergent solutions. Similar to CHS, the negatively charged phospholipids with the serine headgroup (PS) exerted significant stabilizing effects in micelles while uncharged phospholipids were not effective. The purified CB2 reconstituted into lipid bilayers retained functionality for up to several weeks enabling high resolution structural studies of this GPCR at physiologically relevant

  10. Status of the Topside Vary-Chap Ionospheric Model

    NASA Astrophysics Data System (ADS)

    Reinisch, Bodo; Nsumei, Patrick; Huang, Xueqin; Bilitza, Dieter

    Status of the Topside Vary-Chap Ionospheric Model The general alpha-Chapman function for a multi-constituent gas which includes a continuously varying scale height and was therefore dubbed the Vary-Chap function, can present the topside electron density profiles in analytical form. The Vary-Chap profile is defined by the scale height function H(h) and the height and density of the F2 layer peak. By expressing 80,000 ISIS-2 measured topside density profiles as Vary-Chap functions we derived 80,000 scale height functions, which form the basis for the topside density profile modeling. The normalized scale height profiles Hn = H(h)/Hm were grouped according to season, MLAT, and MLT for each 50 km height bin from 200 km to 1400 km, and the median, lower, and upper quartiles for each bin were calculated. Hm is the scale height at the F2 layer peak. The resulting Hn functions are modeled in terms of hyperbolic tangent functions using 5 parameters that are determined by multivariate least squares, including the transition height hT where the scale height gradient has a maximum. These normalized scale height functions, representing the model of the topside electron density profiles from hmF2 to 1,400 km altitude, are independent of hmF2 and NmF2 and can therefore be directly used for the topside Ne profile in IRI. Similarly, this model can extend measured bottomside profiles to the topside, replacing the simple alpha-Chapman function with constant scale height that is currently used for construction of the topside profile in the Digisondes / ARTIST of the Global Ionospheric Radio Observatory (GIRO). It turns out that Hm(top) calculated from the topside profiles is generally several times larger than Hm(bot) derived from the bottomside profiles. This follows necessarily from the difference in the definition of the scale height functions for the topside and bottomside profiles. The diurnal variations of the ratio Hm(top) / Hm(bot) has been determined for different latitudes

  11. The solubilisation of boar sperm membranes by different detergents - a microscopic, MALDI-TOF MS, 31P NMR and PAGE study on membrane lysis, extraction efficiency, lipid and protein composition

    PubMed Central

    2009-01-01

    Background Detergents are often used to isolate proteins, lipids as well as "detergent-resistant membrane domains" (DRMs) from cells. Different detergents affect different membrane structures according to their physico-chemical properties. However, the effects of different detergents on membrane lysis of boar spermatozoa and the lipid composition of DRMs prepared from the affected sperm membranes have not been investigated so far. Results Spermatozoa were treated with the selected detergents Pluronic F-127, sodium cholate, CHAPS, Tween 20, Triton X-100 and Brij 96V. Different patterns of membrane disintegration were observed by light and electron microscopy. In accordance with microscopic data, different amounts of lipids and proteins were released from the cells by the different detergents. The biochemical methods to assay the phosphorus and cholesterol contents as well as 31P NMR to determine the phospholipids were not influenced by the presence of detergents since comparable amounts of lipids were detected in the organic extracts from whole cell suspensions after exposure to each detergent. However, matrix-assisted laser desorption and ionization time-of-flight mass spectrometry applied to identify phospholipids was essentially disturbed by the presence of detergents which exerted particular suppression effects on signal intensities. After separation of the membrane fractions released by detergents on a sucrose gradient only Triton X-100 and sodium cholate produced sharp turbid DRM bands. Only membrane solubilisation by Triton X-100 leads to an enrichment of cholesterol, sphingomyelin, phosphatidylinositol and phosphatidylethanolamine in a visible DRM band accompanied by a selective accumulation of proteins. Conclusion The boar sperm membranes are solubilised to a different extent by the used detergents. Particularly, the very unique DRMs isolated after Triton X-100 exposure are interesting candidates for further studies regarding the architecture of sperm. PMID

  12. Detergent-compatible bacterial amylases.

    PubMed

    Niyonzima, Francois N; More, Sunil S

    2014-10-01

    Proteases, lipases, amylases, and cellulases are enzymes used in detergent formulation to improve the detergency. The amylases are specifically supplemented to the detergent to digest starchy stains. Most of the solid and liquid detergents that are currently manufactured contain alkaline enzymes. The advantages of using alkaline enzymes in the detergent formulation are that they aid in removing tough stains and the process is environmentally friendly since they reduce the use of toxic detergent ingredients. Amylases active at low temperature are preferred as the energy consumption gets reduced, and the whole process becomes cost-effective. Most microbial alkaline amylases are used as detergent ingredients. Various reviews report on the production, purification, characterization, and application of amylases in different industry sectors, but there is no specific review on bacterial or fungal alkaline amylases or detergent-compatible amylases. In this mini-review, an overview on the production and property studies of the detergent bacterial amylases is given, and the stability and compatibility of the alkaline bacterial amylases in the presence of the detergents and the detergent components are highlighted.

  13. Free flow cell electrophoresis using zwitterionic buffer

    NASA Technical Reports Server (NTRS)

    Rodkey, R. Scott

    1990-01-01

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

  14. A zwitterionic macro-crosslinker for durable non-fouling coatings.

    PubMed

    Wang, Wei; Lu, Yang; Xie, Jinbing; Zhu, Hui; Cao, Zhiqiang

    2016-03-28

    A novel zwitterionic macro-crosslinker was developed and applied to fabricate durable non-fouling coatings on a polyurethane substrate. The zwitterionic macro-crosslinker coating exhibited superior durability over the traditional brush polymer coating and was able to retain its non-fouling property even after weeks of shearing in flowing liquid.

  15. Facilitated preparation of bioconjugatable zwitterionic quantum dots using dual-lipid encapsulation.

    PubMed

    Shrake, Robert; Demillo, Violeta G; Ahmadiantehrani, Mojtaba; Zhu, Xiaoshan; Publicover, Nelson G; Hunter, Kenneth W

    2015-01-01

    Zwitterionic quantum dots prepared through incorporated zwitterionic ligands on quantum dot surfaces, are being paid significant attention in biomedical applications because of their excellent colloidal stability across a wide pH and ionic strength range, antifouling surface, good biocompatibility, etc. In this work, we report a dual-lipid encapsulation approach to prepare bioconjugatable zwitterionic quantum dots using amidosulfobetaine-16 lipids, dipalmitoyl-sn-glycero-3-phosphoethanolamine lipids with functional head groups, and CuInS2/ZnS quantum dots in a tetrahydrofuran/methanol/water solvent system with sonication. Amidosulfobetaine-16 is a zwitterionic lipid and dipalmitoyl-sn-glycero-3-phosphoethanolamine, with its functional head, provides bioconjugation capability. Under sonication, tetrahydrofuran/methanol containing amidosulfobetaine-16, dipalmitoyl-sn-glycero-3-phosphoethanolamine, and hydrophobic quantum dots are dispersed in water to form droplets. Highly water-soluble tetrahydrofuran/methanol in droplets is further displaced by water, which induces the lipid self-assembling on hydrophobic surface of quantum dots and thus forms water soluble zwitterionic quantum dots. The prepared zwitterionic quantum dots maintain colloidal stability in aqueous solutions with high salinity and over a wide pH range. They are also able to be conjugated with biomolecules for bioassay with minimal nonspecific binding. Copyright © 2014 Elsevier Inc. All rights reserved.

  16. Laundry detergents: an overview.

    PubMed

    Bajpai, Divya; Tyagi, V K

    2007-01-01

    Nowadays laundry detergents are becoming increasingly popular as they can be metered automatically into the washing machine, impart softness, antistaticness, resiliency to fabrics, mild to eyes and skins and shows good dispersibility in water. Because it is consumed when it is used, the sale of laundry detergent is a rather large business. There are many different kinds or brands of laundry detergent sold, many of them claiming some special qualities as selling points. A Laundry detergent composition is a formulated mixture of raw materials that can be classified into different types based on their properties and function in the final product. The different classes of raw materials are surfactants, builders, bleaching agents, enzymes, and minors which remove dirt, stain, and soil from surfaces or textiles gave them pleasant feel and odour. The physico-chemical properties of surfactants make them suitable for laundry purposes. Laundry detergent has traditionally been a powdered or granular solid, but the use of liquid laundry detergents has gradually increased over the years, and these days use of liquid detergent equals or even exceeds use of solid detergent. This review paper describes the history, composition, types, mechanism, consumption, environmental effects and consumption of laundry detergents.

  17. Expression and solubilization of insect cell-based rabies virus glycoprotein and assessment of its immunogenicity and protective efficacy in mice.

    PubMed

    Ramya, R; Mohana Subramanian, B; Sivakumar, V; Senthilkumar, R L; Sambasiva Rao, K R S; Srinivasan, V A

    2011-10-01

    Rabies is a fatal zoonotic disease of serious public health and economic significance worldwide. The rabies virus glycoprotein (RVG) has been the major target for subunit vaccine development, since it harbors domains responsible for induction of virus-neutralizing antibodies, infectivity, and neurovirulence. The glycoprotein (G) was cloned using the baculovirus expression vector system (BEVS) and expressed in Spodoptera frugiperda (Sf-9) cells. In order to obtain a soluble form of G suitable for experimentation in mice, 18 different combinations of buffers and detergents were evaluated for their ability to solubilize the insect cell membrane-associated G. The combination that involved 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) detergent in lysis buffer 1, formulated with Tris, NaCl, 10% dimethyl sulfoxide (DMSO), and EDTA, gave the highest yield of soluble G, as evidenced by the experimental data. Subsequently, several other parameters, such as the concentration of CHAPS and the duration and temperature of the treatment for the effective solubilization of G, were optimized. The CHAPS detergent, buffered at a concentration of 0.4% to 0.7% (wt/vol) at room temperature (23 to 25°C) for 30 min to 1 h using buffer 1, containing 10% DMSO, resulted in consistently high yields. The G solubilized using CHAPS detergent was found to be immunogenic when tested in mice, as evidenced by high virus-neutralizing antibody titers in sera and 100% protection upon virulent intracerebral challenge with the challenge virus standard (CVS) strain of rabies virus. The results of the mice study indicated that G solubilized with CHAPS detergent retained the immunologically relevant domains in the native conformation, thereby paving the way for producing a cell-free and efficacious subunit vaccine.

  18. Modulation of surgical fibrosis by microbial zwitterionic polysaccharides

    NASA Astrophysics Data System (ADS)

    Ruiz-Perez, Begonia; Chung, Doo R.; Sharpe, Arlene H.; Yagita, Hideo; Kalka-Moll, Wiltrud M.; Sayegh, Mohamed H.; Kasper, Dennis L.; Tzianabos, Arthur O.

    2005-11-01

    Bacterial carbohydrates have long been considered T cell-independent antigens that primarily induce humoral immune responses. Recently, it has been demonstrated that bacterial capsules that possess a zwitterionic charge motif can activate CD4+ T cells after processing and presentation by antigen-presenting cells. Here we show that these zwitterionic polysaccharides can prevent T helper 1-mediated fibrosis by signaling for the release of IL-10 from CD4+ T cells in vivo. IL-10 production by these T cells and their ability to prevent fibrosis is controlled by the inducible costimulator (ICOS)-ICOS ligand pathway. These data demonstrate that the interaction of the zwitterionic polysaccharides with T cells results in modulation of surgical fibrosis in vivo and suggest a previously undescribed approach to "harnessing" T cell function to prevent inflammatory tissue disorders in humans. IL-10 | microbial polysaccharides | inducible costimulator

  19. Smart zwitterionic membranes with on/off behavior for protein transport.

    PubMed

    Su, Yanlei; Zheng, Lili; Li, Chao; Jiang, Zhongyi

    2008-09-25

    Poly(acrylonitrile) (PAN)-based zwitterionic membranes, composed of PAN and poly( N, N-dimethyl- N-methacryloxyethyl- N-(3-sulfopropyl) copolymer, are electrolyte-sensitive smart membranes. The hydrophilicity was increased and protein adsorption was remarkably decreased for the membranes in response to environmental stimuli. FTIR spectroscopic analysis directly provided molecular-level observation of the enhanced dissociation and hydration of zwitterionic sulfobetaine dipoles at higher electrolyte concentrations. The smart PAN-based zwitterionic membranes can close or open channels for protein transport under different NaCl concentrations. The electrolyte-sensitive switch of on/off behavior for protein transport is reversible.

  20. Detergent-mediated protein aggregation

    PubMed Central

    Neale, Chris; Ghanei, Hamed; Holyoake, John; Bishop, Russell E.; Privé, Gilbert G.; Pomès, Régis

    2016-01-01

    Because detergents are commonly used to solvate membrane proteins for structural evaluation, much attention has been devoted to assessing the conformational bias imparted by detergent micelles in comparison to the native environment of the lipid bilayer. Here, we conduct six 500-ns simulations of a system with >600,000 atoms to investigate the spontaneous self assembly of dodecylphosphocholine detergent around multiple molecules of the integral membrane protein PagP. This detergent formed equatorial micelles in which acyl chains surround the protein’s hydrophobic belt, confirming existing models of the detergent solvation of membrane proteins. In addition, unexpectedly, the extracellular and periplasmic apical surfaces of PagP interacted with the headgroups of detergents in other micelles 85 and 60% of the time, respectively, forming complexes that were stable for hundreds of nanoseconds. In some cases, an apical surface of one molecule of PagP interacted with an equatorial micelle surrounding another molecule of PagP. In other cases, the apical surfaces of two molecules of PagP simultaneously bound a neat detergent micelle. In these ways, detergents mediated the non-specific aggregation of folded PagP. These simulation results are consistent with dynamic light scattering experiments, which show that, at detergent concentrations ≥600 mM, PagP induces the formation of large scattering species that are likely to contain many copies of the PagP protein. Together, these simulation and experimental results point to a potentially generic mechanism of detergent-mediated protein aggregation. PMID:23466535

  1. Sulfobetaine as a zwitterionic mediator for 3D hydroxyapatite mineralization

    PubMed Central

    Liu, Pingsheng; Song, Jie

    2013-01-01

    Both positively and negatively charged residues play pivotal roles in recruiting precursor ions or ion clusters, and lowering interfacial energy in natural biomineralization process. Synergistic utilization of opposite charges, however, has rarely been implemented in the design of cytocompatible synthetic scaffolds promoting hydroxyapatite (HA)-mineralization and osteointegration. We report the use of cytocompatible zwitterionic sulfobetaine ligands to enable 3-dimensional in vitro mineralization of HA across covalently crosslinked hydrogels. The overall charge-neutral zwitterionic hydrogel effectively recruited oppositely charged precursor ions while overcame excessive swelling exhibited by anionic and cationic hydrogels under physiological conditions, resulting in denser and structurally well-integrated mineralized composites. Further controls over the size, content, and spatial distribution of the mineral domains within the zwitterionic hydrogel are accomplished by facile adjustments of hydrogel crosslinking densities and the supersaturation rate governing heterogeneous mineral nucleation and growth. These findings should inspire many creative uses of zwitterionic polymers and polymer coatings for skeletal tissue repair and regeneration. PMID:23332320

  2. Antifouling enhancement of polyimide membrane by grafting DEDA-PS zwitterions.

    PubMed

    Zhang, Dong Yan; Xiong, Shu; Shi, Yu Sheng; Zhu, Jun; Hu, Qiao Li; Liu, Jie; Wang, Yan

    2018-05-01

    In order to improve the water flux and antifouling property of polyimide (PI) membrane, zwitterions are grafted on PI membrane surface via a two-step modification route by reactions with N,N-diethylethylenediamine (DEDA) and 1,3-propane sultone (PS) sequentially. The reaction mechanism and physicochemical properties of membranes are confirmed via various characterization techniques. The anti-biofouling performance of the zwitterion-grafted PI membranes is evaluated by bacterial suspension immersion tests in Escherichia coli (E. coli) and staphylococcus aureus (S. aureus) solutions. The antifouling property is assessed via the filtration test using the bovine serum albumin (BSA) and dodecyl trimethyl ammonium bromide (DTAB) aqueous feed solutions. The effect of the reaction time with DEDA in the zwitterion-grafted process on the antifouling property is further investigated systematically. The results show that both the anti-biofouling and antifouling performances of zwitterion-grafted PI membranes are significantly improved. Copyright © 2018 Elsevier Ltd. All rights reserved.

  3. Sulfobetaine as a zwitterionic mediator for 3D hydroxyapatite mineralization.

    PubMed

    Liu, Pingsheng; Song, Jie

    2013-03-01

    Both positively and negatively charged residues play pivotal roles in recruiting precursor ions or ion clusters, and lowering interfacial energy in natural biomineralization process. Synergistic utilization of opposite charges, however, has rarely been implemented in the design of cytocompatible synthetic scaffolds promoting hydroxyapatite (HA)-mineralization and osteointegration. We report the use of cytocompatible zwitterionic sulfobetaine ligands to enable 3-dimensional in vitro mineralization of HA across covalently crosslinked hydrogels. The overall charge-neutral zwitterionic hydrogel effectively recruited oppositely charged precursor ions while overcame excessive swelling exhibited by anionic and cationic hydrogels under physiological conditions, resulting in denser and structurally well-integrated mineralized composites. Further controls over the size, content, and spatial distribution of the mineral domains within the zwitterionic hydrogel are accomplished by facile adjustments of hydrogel crosslinking densities and the supersaturation rate governing heterogeneous mineral nucleation and growth. These findings should inspire many creative uses of zwitterionic polymers and polymer coatings for skeletal tissue repair and regeneration. Copyright © 2012 Elsevier Ltd. All rights reserved.

  4. Stakeholders' Perspectives towards the Use of the Comprehensive Health Assessment Program (CHAP) for Adults with Intellectual Disabilities in Manitoba.

    PubMed

    Shooshtari, Shahin; Temple, Beverley; Waldman, Celeste; Abraham, Sneha; Ouellette-Kuntz, Héléne; Lennox, Nicholas

    2017-07-01

    No standardized tool is used in Canada for comprehensive health assessments of adults with intellectual disabilities. This study was conducted to determine the feasibility of implementing the Comprehensive Health Assessment Program (CHAP) in Manitoba, Canada. This was a qualitative study using a purposive sample of physicians, nurse practitioners, support workers and families. Data were collected through individual interviews and focus groups and were analysed using content analysis. Use of the CHAP was perceived as beneficial for persons with intellectual disabilities. Improved continuity of care was of the reported benefits. Six barriers for the future implementation of the CHAP were identified including the time required to complete the CHAP, and the perceived lack of physicians' willingness to do comprehensive assessments. The future implementation of the CHAP was strongly supported. For its successful implementation, training of healthcare professionals and support staff and change in regulations and policies were recommended. © 2016 John Wiley & Sons Ltd.

  5. AN EXACT METHOD FOR RELATING ZWITTERIONIC MICROSCOPIC TO MACROSCOPIC ACIDITY CONSTANTS

    EPA Science Inventory

    Zwitterions are aqueous solvated molecules simultaneously possessing one negatively and one positively charged site. Although electroneutral, the environmental interaction of zwitterions with other ionic species is likely to differ significantly from the behavior of comparable e...

  6. New zwitterionic monoterpene indole alkaloids from Uncaria rhynchophylla.

    PubMed

    Guo, Qiang; Yang, Hongshuai; Liu, Xinyu; Si, Xiali; Liang, Hong; Tu, Pengfei; Zhang, Qingying

    2018-01-31

    Four new zwitterionic monoterpene indole alkaloids, rhynchophyllioniums A-D (1-4), together with eight known alkaloids (5-12), were isolated from the hook-bearing stems of Uncaria rhynchophylla. Their structures were elucidated by extensive spectroscopic data analysis of MS, 1D and 2D NMR, and ECD, and the zwitterionic forms and absolute configurations of 1 and 2 were unambiguously confirmed by single crystal X-ray diffraction analysis. All the isolates, including the monoterpene indole alkaloids with free C-22 carboxyl group and those with C-22 carboxyl methyl ester, were proved to be naturally coexisting in the herb by LC-MS analysis. This is the first report of monoterpene indole alkaloids that exist in the form of zwitterion. Additionally, the cytotoxic activities of all isolates against A549, HepG2, and MCF-7 cell lines are reported. Copyright © 2018 Elsevier B.V. All rights reserved.

  7. Prevention of bacterial adhesion to zwitterionic biocompatible mesoporous glasses.

    PubMed

    Sánchez-Salcedo, Sandra; García, Ana; Vallet-Regí, María

    2017-07-15

    Novel materials, based on Mesoporous Bioactive Glasses (MBGs) in the ternary system SiO 2 -CaO-P 2 O 5 , decorated with (3-aminopropyl)triethoxysilane (APTES) and subsequently with amino acid Lysine (Lys), by post-grafting method on the external surface of the glasses (named MBG-NH 2 and MBG-Lys), are reported. The surface functionalization with organic groups did not damage the mesoporous network and their structural and textural properties were also preserved despite the high solubility of MBG matrices. The incorporation of Lys confers a zwitterionic nature to these MBG materials due to the presence of adjacent amine and carboxylic groups in the external surface. At physiologic pH, this coexistence of basic amine and carboxilic acid groups from anchored Lys provided zero surface charge named zwitterionic effect. This behaviour could give rise to potential applications of antibacterial adhesion. Therefore, in order to assess the influence of zwitterionic nature in in vitro bacterial adhesion, studies were carried out with Staphylococcus aureus. It was demonstrated that the efficient interaction of these zwitterionic pairs onto the MBG surfaces reduced bacterial adhesion up to 99.9% compared to bare MBGs. In order to test the suitability of zwitterionic MBGs materials as bone grafts, their cytocompatibility was investigated in vitro with MC3T3-E1 preosteoblasts. These findings suggested that the proposed surface functionalization strategy provided MBG materials with notable antibacterial adhesion properties, hence making these materials promising candidates for local bone infection therapy. The present research work is focused in finding a preventive treatment of bone infection based on Mesoporous Bioactive Glasses (MBGs) with antibacterial adhesion properties obtained by zwitterionic surface modification. MBGs exhibit unique nanostructural, textural and bioactive characteristics. The novelty and originality of this manuscript is based on the design and

  8. [Determination of the detergent efficiency of the detergents disinfecting for surfaces].

    PubMed

    Santucci, R; Kuntzmann, X; Mesli, N; Meunier, O

    2009-01-01

    Since the disinfecting activity of disinfectants is evaluated by standards, the intrinsic detergent activity is not easily quantifiable and no standard have been suggested yet. Beyond the physicochemical parameters like wettability or foaming presented by the manufacturers, it appears necessary to us to objectively measure the real effect of the detergent agent. The objective of our work is to propose a simple, fast and reproducible method to evaluate detersive activity of the disinfecting detergents. We measured three factors (total amount of extracted bacteria, extraction efficiency and slope of extracting curve) by using Rodac prints technique on two different supports (PVC, stainless steel) that have been contaminated by either E. coli or S. aureus. An increasing mark from 1 to 6 is given to each of these factors in case of statistically differences. The three factors allowed us to calculate a "Specific Index of detersion" (SI) for each germ/support couple (3 to 18). Addition of the marks given to each couple for each disinfecting detergent allowed to calculate a "Globally Index of detersion" (GI) (9 to 72). We tested 4 commercialised disinfecting detergents: Surfanios, Aniosurf, Major C100 and Ecodiol. All detergents may be classified according to their effectiveness on a bacterium/support couple (value of the SI). This enlights a specific spectrum for each disinfecting detergents. As a result, Ecodiol seems to be the most effective deterging agent on 3 of the 4 germ/support couples (S. aureus/PVC, E. coli/PVC and E. coli/stainless steel), whereas Aniosurf is most effective on the S. aureus/stainless steel couple. The GI is very useful to choose the best compromise between activities for all situations. GI rankings of the tested agents were as follows: water < Aniosurf < Surfanios < neutralizing < Major C100 < Ecodiol. This experimental model will be used to test and compare the intrinsic detergent activities of other commercialised products which are usually

  9. Use of a crossed high alcohol preferring (cHAP) mouse model with the NIAAA-model of chronic-binge ethanol intake to study liver injury.

    PubMed

    Thompson, Kyle J; Nazari, Shayan S; Jacobs, W Carl; Grahame, Nicholas J; McKillop, Iain H

    2017-11-01

    This study sought to compare mice bred to preferentially consume high amounts of alcohol (crossed-high alcohol preferring, cHAP) to c57BL/6 (C57) mice using a chronic-binge ethanol ingestion model to induce alcoholic liver disease (ALD). Male C57 and cHAP mice were randomized to a Lieber-DeCarli control (LDC) diet, Lieber-DeCarli 5% (v/v) ethanol (LDE) diet or free-choice between 10% (v/v) ethanol in drinking water (EtOH-DW) and DW. After 4 weeks mice were gavaged with either 9 g/kg maltose-dextrin (LDC+MD) or 5 g/kg EtOH (LDE+Binge, EtOH-DW+Binge). Nine hours later tissue and serum were collected and analyzed. cHAP mice on EtOH-DW consumed significantly more ethanol than cHAP or C57 mice maintained on LDE. However, cHAP and C57 mice on the LDE+Binge regiment had greater hepatosteatosis and overall degree of liver injury compared to EtOH-DW+Binge. Changes in pro-inflammatory gene expression was more pronounced in cHAP mice than C57 mice. Analysis of liver enzymes revealed a robust induction of CYP2E1 in C57 and cHAP mice maintained on EtOH-DW+Binge or LDE+Binge. However, while C57 mice exhibited higher basal hepatic glutathione than cHAP mice, these mice appeared more susceptible to oxidative stress following LDE+Binge than cHAP counterparts. Despite cHAP mice consuming more total ethanol prior to gavage when maintained on EtOH-DW, LDE followed by gavage created a more severe model of ALD in both C57 and cHAP mice. These data suggest factors other than total amount of alcohol consumed affect degree of ALD development in the chronic-binge model in cHAP mice. cHAP mice voluntarily consume high amounts of ethanol and exhibited hepatic injury when subject to chronic-binge ethanol feeding with the Lieber-DeCarli diet. However, hepatic injury was reduced in cHAP mice in a chronic-binge model following voluntary high ethanol consumption in drinking water. © The Author 2017. Medical Council on Alcohol and Oxford University Press. All rights reserved.

  10. Carica papaya lipase: a naturally immobilized enzyme with interesting biochemical properties.

    PubMed

    Abdelkafi, Slim; Barouh, Nathalie; Fouquet, Benjamin; Fendri, Imen; Pina, Michel; Scheirlinckx, Frantz; Villeneuve, Pierre; Carrière, Frédéric

    2011-03-01

    Triacylglycerol (TAG) lipases have been thoroughly characterized in mammals and microorganisms, whereas very little is known about plant TAG lipases. The lipolytic activity occurring in all the laticies is known to be associated with sedimentable particles, and all attempts to solubilize the lipolytic activity of Carica papaya latex have been unsuccessful so far. However, some of the biochemical properties of the lipase from Carica papaya latex (CPL) were determined from the insoluble fraction of the latex. The activity was optimum at a temperature of 37°C and a pH of 9.0, and the specific activities of CPL were found to be 2,000 ± 185 and 256 ± 8 U/g when tributyrin and olive oil were used as substrates, respectively. CPL was found to be active in the absence of any detergent, whereas many lipases require detergent to prevent the occurrence of interfacial denaturation. CPL was inactive in the presence of micellar concentrations of Triton X-100, sodium dodecyl sulfate (SDS) and tetradecyl trimethylammonium bromide (TTAB), and still showed high levels of activity in the presence of sodium taurodeoxycholate (NaTDC) and the zwitterionic Chaps detergent. The effects of various proteases on the lipolytic activity of CPL were studied, and CPL was found to be resistant to treatment with various enzymes, except in the presence of trypsin. All these properties suggest that CPL may be a good candidate for various biotechnological applications.

  11. Surface zwitterionization: Effective method for preventing oral bacterial biofilm formation on hydroxyapatite surfaces

    NASA Astrophysics Data System (ADS)

    Lee, Myoungjin; Kim, Heejin; Seo, Jiae; Kang, Minji; Kang, Sunah; Jang, Joomyung; Lee, Yan; Seo, Ji-Hun

    2018-01-01

    In this study, we conducted surface zwitterionization of hydroxyapatite (HA) surfaces by immersing them in the zwitterionic polymer solutions to provide anti-bacterial properties to the HA surface. Three different monomers containing various zwitterionic groups, i.e., phosphorylcholine (PC), sulfobetaine (SB), and carboxybetaine (CB), were copolymerized with the methacrylic monomer containing a Ca2+-binding moiety, using the free radical polymerization method. As a control, functionalization of the copolymer containing the Ca2+-binding moiety was synthesized using a hydroxy group. The stable immobilization of the zwitterionic functional groups was confirmed by water contact angle analysis and X-ray photoelectron spectroscopy (XPS) measurement conducted after the sonication process. The zwitterionized HA surface showed significantly decreased protein adsorption, whereas the hydroxyl group-coated HA surface showed limited efficacy. The anti-bacterial adhesion property was confirmed by conducting Streptococcus mutans (S. mutans) adhesion tests for 6 h and 24 h. When furanone C-30, a representative anti-quorum sensing molecule for S. mutans, was used, only a small amount of bacteria adhered after 6 h and the population did not increase after 24 h. In contrast, zwitterionized HA surfaces showed almost no bacterial adhesion after 6 h and the effect was retained for 24 h, resulting in the lowest level of oral bacterial adhesion. These results confirm that surface zwitterionization is a promising method to effectively prevent oral bacterial adhesion on HA-based materials.

  12. Molecular level studies on interfacial hydration of zwitterionic and other antifouling polymers in situ.

    PubMed

    Leng, Chuan; Sun, Shuwen; Zhang, Kexin; Jiang, Shaoyi; Chen, Zhan

    2016-08-01

    Antifouling polymers have wide applications in biomedical engineering and marine industry. Recently, zwitterionic materials have been reported as promising candidates for antifouling applications, while strong hydration is believed to be the key antifouling mechanism. Zwitterionic materials can be designed with various molecular structures, which affect their hydration and antifouling performance. Although strong hydration has been proposed to occur at the material surfaces, probing the solid material/water interfaces is challenging with traditional analytical techniques. Here in this review, we will review our studies on surface hydration of zwitterionic materials and other antifouling materials by using sum frequency generation (SFG) vibrational spectroscopy, which provides molecular understanding of the water structures at various material surfaces. The materials studied include zwitterionic polymer brushes with different molecular structures, amphiphilic polymers with zwitterionic groups, uncharged hydrophilic polymer brushes, amphiphilic polypeptoids, and widely used antifouling material poly(ethylene glycol). We will compare the differences among zwitterionic materials with various molecular structures as well as the differences between antifouling materials and fouling surfaces of control samples. We will also discuss the effects of pH and biological molecules like proteins on the surface hydration of the zwitterionic materials. Using SFG spectroscopy, we have measured the hydration layers of antifouling materials and found that strong hydrogen bonds are key to the formation of strong hydration layers preventing protein fouling at the polymer interfaces. Antifouling polymers have wide applications in biomedical engineering and marine industry. Recently, zwitterionic materials have been reported as promising candidates for antifouling applications, while strong hydration is believed to be the key antifouling mechanism. However, zwitterionic materials can be

  13. Molecular design of high performance zwitterionic liquids for enhanced heavy-oil recovery processes.

    PubMed

    Martínez-Magadán, J M; Cartas-Rosado, A R; Oviedo-Roa, R; Cisneros-Dévora, R; Pons-Jiménez, M; Hernández-Altamirano, R; Zamudio-Rivera, L S

    2018-03-01

    Branched gemini zwitterionic liquids, which contain two zwitterionic moieties of linked quaternary-ammonium and carboxylate groups, are proposed as chemicals to be applied in the Enhanced Oil Recovery (EOR) from fractured carbonate reservoirs. The zwitterionic moieties are bridged between them through an alkyl chain containing 12 ether groups, and each zwitterionic moiety has attached a long alkyl tail including a CC double bond. A theoretical molecular mechanism over which EOR could rest, consisting on both the disaggregation of heavy oil and the reservoir-rock wettability alteration, was suggested. Results show that chemicals can both reduce the viscosity and remove heavy-oil molecules from the rock surface. Copyright © 2018. Published by Elsevier Inc.

  14. Study of the nanoporous CHAP photoluminiscence for developing the precise methods of early caries detection

    NASA Astrophysics Data System (ADS)

    Goloshchapov, D.; Seredin, P.; Minakov, D.; Domashevskaya, E.

    2018-02-01

    This paper deals with the luminescence characteristics of an analogue of the mineral component of dental enamel of the nanocrystalline B-type carbonate-substituted hydroxyapatite (CHAP) with 3D defects (i.e. nanopores of ∼2-5 nm) on the nanocrystalline surface. The laser-induced luminescence of the synthesized CHAP samples was in the range of ∼515 nm (∼2.4 eV) and is due to CO3 groups replacing the PO4 group. It was found that the intensity of the luminescence of the CHAP is caused by structurally incorporated CO3 groups in the HAP structure. Furthermore, the intensity of the luminescence also decreases as the number of the above intracentre defects (CO3) in the apatite structure declines. These results are potentially promising for developing the foundations for precise methods for the early detection of caries in human solid dental tissue.

  15. Zwitteration: Coating Surfaces with Zwitterionic Functionality to Reduce Nonspecific Adsorption

    PubMed Central

    2015-01-01

    Coating surfaces with thin or thick films of zwitterionic material is an effective way to reduce or eliminate nonspecific adsorption to the solid/liquid interface. This review tracks the various approaches to zwitteration, such as monolayer assemblies and polymeric brush coatings, on micro- to macroscopic surfaces. A critical summary of the mechanisms responsible for antifouling shows how zwitterions are ideally suited to this task. PMID:24754399

  16. A zwitterionic gel electrolyte for efficient solid-state supercapacitors

    PubMed Central

    Peng, Xu; Liu, Huili; Yin, Qin; Wu, Junchi; Chen, Pengzuo; Zhang, Guangzhao; Liu, Guangming; Wu, Changzheng; Xie, Yi

    2016-01-01

    Gel electrolytes have attracted increasing attention for solid-state supercapacitors. An ideal gel electrolyte usually requires a combination of advantages of high ion migration rate, reasonable mechanical strength and robust water retention ability at the solid state for ensuring excellent work durability. Here we report a zwitterionic gel electrolyte that successfully brings the synergic advantages of robust water retention ability and ion migration channels, manifesting in superior electrochemical performance. When applying the zwitterionic gel electrolyte, our graphene-based solid-state supercapacitor reaches a volume capacitance of 300.8 F cm−3 at 0.8 A cm−3 with a rate capacity of only 14.9% capacitance loss as the current density increases from 0.8 to 20 A cm−3, representing the best value among the previously reported graphene-based solid-state supercapacitors, to the best of our knowledge. We anticipate that zwitterionic gel electrolyte may be developed as a gel electrolyte in solid-state supercapacitors. PMID:27225484

  17. Zwitterionic Group VIII transition metal initiators supported by olefin ligands

    DOEpatents

    Bazan, Guillermo C [Goleta, CA; Chen, Yaofeng [Shanghai, CN

    2011-10-25

    A zwitterionic Group VIII transition metal complex containing the simple and relatively small 3-(arylimino)-but-1-en-2-olato ligand that catalyzes the formation of polypropylene and high molecular weight polyethylene. A novel feature of this catalyst is that the active species is stabilized by a chelated olefin adduct. The present invention also provides methods of polymerizing olefin monomers using zwitterionic catalysts, particularly polypropylene and high molecular weight polyethylene.

  18. CHAP-2 heat-transfer analysis of the Fort St. Vrain reactor core

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

    Kotas, J.F.; Stroh, K.R.

    1983-01-01

    The Los Alamos National Laboratory is developing the Composite High-Temperature Gas-Cooled Reactor Analysis Program (CHAP) to provide advanced best-estimate predictions of postulated accidents in gas-cooled reactor plants. The CHAP-2 reactor-core model uses the finite-element method to initialize a two-dimensional temperature map of the Fort St. Vrain (FSV) core and its top and bottom reflectors. The code generates a finite-element mesh, initializes noding and boundary conditions, and solves the nonlinear Laplace heat equation using temperature-dependent thermal conductivities, variable coolant-channel-convection heat-transfer coefficients, and specified internal fuel and moderator heat-generation rates. This paper discusses this method and analyzes an FSV reactor-core accident thatmore » simulates a control-rod withdrawal at full power.« less

  19. Cardiovascular Health Awareness Program (CHAP): a community cluster-randomised trial among elderly Canadians.

    PubMed

    Kaczorowski, Janusz; Chambers, Larry W; Karwalajtys, Tina; Dolovich, Lisa; Farrell, Barbara; McDonough, Beatrice; Sebaldt, Rolf; Levitt, Cheryl; Hogg, William; Thabane, Lehana; Tu, Karen; Goeree, Ron; Paterson, J Michael; Shubair, Mamdouh; Gierman, Tracy; Sullivan, Shannon; Carter, Megan

    2008-06-01

    High blood pressure is an important and modifiable cardiovascular disease risk factor that remains under-detected and under-treated. Community-level interventions that address high blood pressure and other modifiable risk factors are a promising strategy to improve cardiovascular health in populations. The present study is a community cluster-randomised trial testing the effectiveness of CHAP (Cardiovascular Health Awareness Program) on the cardiovascular health of older adults. Thirty-nine mid-sized communities in Ontario, Canada were stratified by geographic location and size of the population aged >or=65 years and randomly allocated to receive CHAP or no intervention. In CHAP communities, residents aged >or=65 years were invited to attend cardiovascular risk assessment sessions held in pharmacies over 10 weeks in Fall, 2006. Sessions included blood pressure measurement and feedback to family physicians. Trained volunteers delivered the program with support from pharmacists, community nurses and local organisations. The primary outcome measure is the relative change in the mean annual rate of hospital admission for acute myocardial infarction, congestive heart failure and stroke (composite end-point) among residents aged >or=65 years in intervention and control communities, using routinely collected, population-based administrative health data. This paper highlights considerations in design, implementation and evaluation of a large-scale, community-wide cardiovascular health promotion initiative.

  20. Universal Coatings Based on Zwitterionic-Dopamine Copolymer Microgels.

    PubMed

    Vatankhah-Varnosfaderani, Mohammad; Hu, Xiaobo; Li, Qiaoxi; Adelnia, Hossein; Ina, Maria; Sheiko, Sergei S

    2018-06-05

    Multifunctional coatings that adhere to chemically distinct substrates are vital in many industries, including automotive, aerospace, shipbuilding, construction, petrochemical, biomedical, and pharmaceutical. We design well-defined, nearly monodisperse microgels that integrate hydrophobic dopamine methacrylamide monomers and hydrophilic zwitterionic monomers. The dopamine functionalities operate as both intraparticle cross-linkers and interfacial binders, respectively providing mechanical strength of the coatings and their strong adhesion to different substrates. In tandem, the zwitterionic moieties enable surface hydration to empower antifouling and antifogging properties. Drop-casting of microgel suspensions in ambient as well as humid environments facilitates rapid film formation and tunable roughness through regulation of cross-linking density and deposition conditions.

  1. Facile surface modification of glass with zwitterionic polymers for improving the blood compatibility

    NASA Astrophysics Data System (ADS)

    Zhang, Lingling; Chen, Xiaojuan; Liu, Pingsheng; Wang, Jing; Zhu, Haomiao; Li, Li

    2018-06-01

    A facile procedure to modify glass film with zwitterionic polymers for improving the blood compatibility was introduced. The glass slides were first silanized with 3-methacryloxypropyltrimethoxysilane (MPT) to generate methacrylate groups on the surface. Then, N, N’-dimethyl-N-methacryloxyethyl-N-(3-sulfopropyl) ammonium (DMMSA), a sulfobetaine zwitterionic monomer, was polymerized on the silanized glass substrates by free-radical polymerization in order to graft the zwitterionic polymers onto the substrates. X-ray Photoelectron Spectroscopy (XPS), water contact angle, scanning electron microscope (SEM) and atomic force microscopy (AFM) were utilized to analyze the surface properties of the grafted glass. The blood compatibility of the grafted glass was verified by whole blood contacting and platelet adhesion experiments in vitro. The results showed that the zwitterionic polymers were successfully grafted on the glass surface, and consequently significantly inhibited the platelet adhesion and whole blood cell attachment.

  2. Stable isotope (C, O) and monovalent cation fractionation upon synthesis of carbonate-bearing hydroxyl apatite (CHAP) via calcite transformation

    NASA Astrophysics Data System (ADS)

    Böttcher, Michael E.; Schmiedinger, Iris; Wacker, Ulrike; Conrad, Anika C.; Grathoff, Georg; Schmidt, Burkhard; Bahlo, Rainer; Gehlken, Peer-L.; Fiebig, Jens

    2016-04-01

    Carbonate-bearing hydroxyl-apatite (CHAP) is of fundamental and applied interest to the (bio)geochemical, paleontological, medical and material science communities, since it forms the basic mineral phase in human and animal teeth and bones. In addition, it is found in non-biogenic phosphate deposits. The stable isotope and foreign element composition of biogenic CHAP is widely used to estimate the formation conditions. This requires careful experimental calibration under well-defined boundary conditions. Within the DFG project EXCALIBOR, synthesis of carbonate-bearing hydroxyapatite was conducted via the transformation of synthetic calcite powder in aqueous solution as a function of time, pH, and temperature using batch-type experiments. The aqueous solution was analyzed for the carbon isotope composition of dissolved inorganic carbonate (gas irmMS), the oxygen isotope composition of water (LCRDS), and the cationic composition. The solid was characterized by powder X-ray diffraction, micro Raman and FTIR spectroscopy, SEM-EDX, elemental analysis (EA, ICP-OES) and gas irmMS. Temperature was found to significantly impact the transformation rate of calcite to CHAP. Upon complete transformation, CHAP was found to contain up to 5% dwt carbonate, depending on the solution composition (e.g., pH), both incorporated on the A and B type position of the crystal lattice. The oxygen isotope fractionation between water and CHAP decreased with increasing temperature with a tentative slope shallower than those reported in the literature for apatite, calcite or aragonite. In addition, the presence of dissolved NH4+, K+ or Na+ in aqueous solution led to partial incorporation into the CHAP lattice. How these distortions of the crystal lattice may impact stable isotope discrimination is subject of future investigations.

  3. Affinity Chromatography in Nonionic Detergent Solutions

    NASA Astrophysics Data System (ADS)

    Robinson, Jack B.; Strottmann, James M.; Wick, Donald G.; Stellwagen, Earle

    1980-10-01

    Anionic dye affinity chromatography is commonly unproductive in the presence of nonionic detergents used to extract particulate proteins. Using lactate dehydrogenase as a model protein, Cibacron blue F3GA as a model dye, and Triton X-100 as a model detergent, we find that the dye is encapsulated in nonionic detergent micelles, rendering the dye incapable of ligation with the enzyme. However, the dye can be liberated from the micelles without altering the nonionic detergent concentration by addition of an anionic detergent, such as deoxycholate or sodium dodecyl sulfate, forming mixed anionic/nonionic micelles that displace the anionic dye. Encapsulation of the anionic detergents prevents their activity as protein denaturants. These observations have been successfully translated to the dye affinity chromatography of a detergent extract of brain particulate cyclic nucleotide phosphodiesterase.

  4. Characterization of Structure, Dynamics, and Detergent Interactions of the Anti-HIV Chemokine Variant 5P12-RANTES

    PubMed Central

    Wiktor, Maciej; Hartley, Oliver; Grzesiek, Stephan

    2013-01-01

    RANTES (CCL5) is a chemokine that recruits immune cells to inflammatory sites by interacting with the G-protein coupled receptor CCR5, which is also the primary coreceptor used together with CD4 by HIV to enter and infect target cells. Ligands of CCR5, including chemokines and chemokine analogs, are capable of blocking HIV entry, and studies of their structures and interactions with CCR5 will be key to understanding and optimizing HIV inhibition. The RANTES derivative 5P12-RANTES is a highly potent HIV entry inhibitor that is being developed as a topical HIV prevention agent (microbicide). We have characterized the structure and dynamics of 5P12-RANTES by solution NMR. With the exception of the nine flexible N-terminal residues, 5P12-RANTES has the same structure as wild-type RANTES but unlike the wild-type, does not dimerize via its N-terminus. To prepare the ground for interaction studies with detergent-solubilized CCR5, we have also investigated the interaction of RANTES and 5P12-RANTES with various commonly used detergents. Both RANTES variants are stable in Cymal-5, DHPC, Anzergent-3-12, dodecyltrimethylammonium chloride, and a DDM/CHAPS/CHS mixture. Fos-Cholines, dodecyldimethylglycine, and sodium dodecyl-sulfate denature both RANTES variants at low pH, whereas at neutral pH the stability is considerably higher. The onset of Fos-Choline-12-induced denaturation and the denatured state were characterized by circular dichroism and NMR. The detergent interaction starts below the critical micelle concentration at a well-defined mixed hydrophobic/positive surface region of the chemokine, which overlaps with the dimer interface. An increase of Fos-Choline-12 concentration above the critical micelle concentration causes a transition to a denatured state with a high α-helical content. PMID:24314089

  5. Crystal engineering of a zwitterionic drug to neutral cocrystals: a general solution for floxacins.

    PubMed

    Gunnam, Anilkumar; Suresh, Kuthuru; Ganduri, Ramesh; Nangia, Ashwini

    2016-10-18

    The transformation of zwitterionic Sparfloxacin (SPX) to the neutral form is achieved by cocrystallization. Neutral forms of drugs are important for higher membrane permeability, while zwitterions are more soluble in water. The twin advantages of higher solubility/dissolution rate and good stability of neutral SPX are achieved in a molecular cocrystal compared to its zwitterionic SPX hydrate. The amine-phenol supramolecular synthon drives cocrystal formation, with the paraben ester acting as a "proton migrator" for the ionic to neutral transformation.

  6. Microscopic insight into the DNA condensation process of a zwitterion-functionalized polycation.

    PubMed

    Sun, Hui; Zhou, Li; Chen, Xiaolu; Han, Xia; Wang, Rui; Liu, Honglai

    2016-11-01

    Zwitterion-functionalized polycations are ideal gene carriers with long circulation, high cellular uptaking and low cell viability. However, the trade-off between the DNA condensation efficiency and the cell viability must be addressed. The purpose of this study is to provide a microscopic insight into the DNA condensation process and to explore the effect of a zwitterionic block of zwitterion-functionalized polycation, which is of great significance in designing novel gene delivery systems. Poly[2-(dimethylamino)ethyl methacrylate-b-(sulfobetaine methacrylate)] (PDMAEMA-b-PSBMA) copolymers were synthesized and used as the model systems. Different from the conventional concept that the PSBMA zwitterionic block act only as the "stealthy" groups, the subtle differences in physical and colloidal characteristics between the polycation/DNA polyplexes show that the PSBMA segment is capable of wrapping DNA attributed to the quaternary ammonium cations, without compromising the DNA condensation capability. On the other hand, the incorporation of PSBMA block reduces the surface charge of the polyplexes, which substantially result in the inefficient transfection and the reduced cytotoxicity. © 2016 Wiley Periodicals, Inc.

  7. Imidazoline fuel detergents

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

    Bonazza, B. R.; Holtz, H. D.

    1981-01-27

    A detergent additive imidazoline prepared by reacting carboxylic acid with polyamine is combined into fuel for an internal combustion engine or lubricating oil as a composition suitable for reducing deposits in an internal combustion engine. In an embodiment of the invention, the imidazoline is further combined with a sulfonic acid to obtain a fuel detergent of improved operability.

  8. Harmful effect of detergents on lipase.

    PubMed

    Fatima, Sadaf; Ajmal, Rehan; Badr, Gamal; Khan, Rizwan H

    2014-11-01

    In order to study effects of detergents at molecular level, we have done activity measurements of wheat germ lipase in increasing concentration of some commercial detergents. Conformational changes in protein structure using circular dichroism and fluorescence spectroscopy were studied in increasing concentration of sodium dodecyl sulfate. Our study proves that detergents may lead to loss of enzymatic activity and structure of plant enzymes. Since detergents are common source of pollution in water bodies and the water from these resources can be used in fields, our study may prove helpful in creating awareness about harmful action of detergents.

  9. Efficient Preparation of Super Antifouling PVDF Ultrafiltration Membrane with One Step Fabricated Zwitterionic Surface.

    PubMed

    Zhao, Xinzhen; He, Chunju

    2015-08-19

    On the basis of the excellent fouling resistance of zwitterionic materials, the super antifouling polyvinylidene fluoride (PVDF) membrane was efficiently prepared though one-step sulfonation of PVDF and polyaniline blend membrane in situ. The self-doped sulfonated polyaniline (SPANI) was generated as a novel zwitterionic polymer to improve the antifouling property of PVDF ultrafiltration membrane used in sewage treatment. Surface attenuated total reflection Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, surface zeta potential, and water contact angle demonstrated the successful fabrication of zwitterionic interface by convenient sulfonation modification. The static adsorption fouling test showed the quantified adsorption mass of bovine serum albumin (BSA) pollutant on the PVDF/SPANI membrane surface decreases to 3(±2) μg/cm(2), and the water flux recovery ratio (FRR) values were no less than 95% for the three model pollutants of BSA, sodium alginate (SA), and humic acid (HA), which were corresponding hydrophobic, hydrophilic, and natural pollutants in sewage, respectively. This Research Article demonstrated the antifouling advantages of zwitterionic SPANI and aimed to provide a simple method for the large scale preparation of zwitterionic antifouling ultrafiltration membranes.

  10. A novel approach for synthesis of zwitterionic polyurethane coating with protein resistance.

    PubMed

    Wang, Chunhua; Ma, Chunfeng; Mu, Changdao; Lin, Wei

    2014-11-04

    We have developed a novel approach to introduce zwitterions into polyurethane for the preparation of antibiofouling coating. First, the thiol-ene click reaction between 2-(dimethylamino)ethyl methacrylate (DMAEMA) and 3-mercapto-1,2-propanediol (TPG) is used to synthesize dihydroxy-terminated DMAEMA (DMA(OH)2) under UV catalysis. The product has been proved by gel permeation chromatography (GPC), Fourier transform infrared spectrum (FT-IR), proton nuclear magnetic resonance ((1)H NMR), and high resolution mass spectrometry (HRMS). DMA(OH)2 is then incorporated into polyurethane as side groups by polyaddition with diisocyanate and further reacts with 1,3-propane sultone to obtain the zwitterionic polyurethanes. The presence of sulfobetaine zwitterions side groups has been demonstrated by FT-IR and X-ray photoelectron spectroscopy (XPS). Thermal analysis indicates that the thermal stability is decreased with the increasing content of zwitterionions. The antibiofouling property of polyurethanes has been investigated by the measurement of adsorption of fibrinogen, bovine serum albumin (BSA), and lysozyme on the polyurethanes surface using quartz crystal microbalance with dissipation (QCM-D). The results show that the polyurethane coatings exhibit effective nonspecific protein resistance at higher content of zwitterionic side groups.

  11. Zwitterionic versus canonical amino acids over the various defects in zeolites: A two-layer ONIOM calculation

    PubMed Central

    Yang, Gang; Zhou, Lijun

    2014-01-01

    Defects are often considered as the active sites for chemical reactions. Here a variety of defects in zeolites are used to stabilize zwitterionic glycine that is not self-stable in gas phase; in addition, effects of acidic strengths and zeolite channels on zwitterionic stabilization are demonstrated. Glycine zwitterions can be stabilized by all these defects and energetically prefer to canonical structures over Al and Ga Lewis acidic sites rather than Ti Lewis acidic site, silanol and titanol hydroxyls. For titanol (Ti-OH), glycine interacts with framework Ti and hydroxyl sites competitively, and the former with Lewis acidity predominates. The transformations from canonical to zwitterionic glycine are obviously more facile over Al and Ga Lewis acidic sites than over Ti Lewis acidic site, titanol and silanol hydroxyls. Charge transfers that generally increase with adsorption energies are found to largely decide the zwitterionic stabilization effects. Zeolite channels play a significant role during the stabilization process. In absence of zeolite channels, canonical structures predominate for all defects; glycine zwitterions remain stable over Al and Ga Lewis acidic sites and only with synergy of H-bonding interactions can exist over Ti Lewis acidic site, while automatically transform to canonical structures over silanol and titanol hydroxyls. PMID:25307449

  12. Catalytic copolymerization of CO and ethylene with a charge neutral palladium(II) zwitterion.

    PubMed

    Lu, Connie C; Peters, Jonas C

    2002-05-15

    The synthesis of a zwitterionic Pd(II) complex supported by an anionic bis(phosphino)borate ligand, Ph(2)B(CH(2)PPh(2))(2) (abbreviated as [Ph(2)BP(2)]), is reported. The new complex, [Ph(2)BP(2)]PdMe(THF), is active for CO and ethylene copolymerization. The copolymerization activity and polyketone molecular weight for the neutral, zwitterionic system are compared with those for the cationic systems [R(2)E(CH(2)PPh(2))(2)PdMe(THF)][B(C(6)F(5))(4)] where ER(2) = SiPh(2) and CH(2). Surprisingly, the more electron rich zwitterionic system is a catalyst of activity comparable to that of the more conventional cationic systems.

  13. Water-soluble polymers bearing phosphorylcholine group and other zwitterionic groups for carrying DNA derivatives.

    PubMed

    Lin, Xiaojie; Ishihara, Kazuhiko

    2014-01-01

    Water-soluble polymers with equal positive and negative charges in the same monomer unit, such as the phosphorylcholine group and other zwitterionic groups, exhibit promising potential in gene delivery with appreciable transfection efficiency, compared with the traditional poly(ethylene glycol)-based polycation-gene complexes. These zwitterionic polymers with various architectural structures and properties have been synthesized by various polymerization methods, such as conventional radical polymerization, atom-transfer radical-polymerization, reversible addition-fragmentation chain-transfer polymerization, and nitroxide-mediated radical polymerization. These techniques have been used to efficiently facilitate gene therapy by fabrication of non-viral vectors with high cytocompatibility, large gene-carrying capacity, effective cell-membrane permeability, and in vivo gene-loading/releasing functionality. Zwitterionic polymer-based gene delivery vectors systems can be categorized into soluble-polymer/gene mixing, molecular self-assembly, and polymer-gene conjugation systems. This review describes the preparation and characterization of various zwitterionic polymer-based gene delivery vectors, specifically water-soluble phospholipid polymers for carrying gene derivatives.

  14. Design of the Chicago Health and Aging Project (CHAP).

    PubMed

    Bienias, Julia L; Beckett, Laurel A; Bennett, David A; Wilson, Robert S; Evans, Denis A

    2003-10-01

    The design of the Chicago Health and Aging Project (CHAP) is described. CHAP is a longitudinal population study of common chronic health problems of older persons, especially of risk factors for incident Alzheimer's disease, in a biracial neighborhood of the south side of Chicago. Special attention is given to three unusual design features of the study. One feature is that clinical evaluation for Alzheimer's disease is confined to a stratified random sample of all participants. This feature results in substantial cost savings and substantially less bias than screening approaches but has the disadvantages of adding analytic complexity and requiring the use of indirect means to identify a disease-free cohort for the development of incident Alzheimer's disease. The second unusual feature is efficiently combining in analyses the successive independent multiple samples that are drawn, one from each data collection cycle. The third unusual feature is entering successive age cohorts of community residents into the study as they attain 65 years of age. This has the advantages of enhancing direct investigation of the effect of age on the action of risk factors for Alzheimer's disease and direct examination of cohort effects. The interaction of these features is described, especially as they pertain to a study in which data are collected in successive waves. The results from these waves must be combined for effective analysis of the relation among risk factors and incident disease.

  15. Zwitterionic PEG-PC Hydrogels Modulate the Foreign Body Response in a Modulus-Dependent Manner.

    PubMed

    Jansen, Lauren E; Amer, Luke D; Chen, Esther Y-T; Nguyen, Thuy V; Saleh, Leila S; Emrick, Todd; Liu, Wendy F; Bryant, Stephanie J; Peyton, Shelly R

    2018-05-15

    Reducing the foreign body response (FBR) to implanted biomaterials will enhance their performance in tissue engineering. Poly(ethylene glycol) (PEG) hydrogels are increasingly popular for this application due to their low cost, ease of use, and the ability to tune their compliance via molecular weight and cross-linking densities. PEG hydrogels can elicit chronic inflammation in vivo, but recent evidence has suggested that extremely hydrophilic, zwitterionic materials and particles can evade the immune system. To combine the advantages of PEG-based hydrogels with the hydrophilicity of zwitterions, we synthesized hydrogels with comonomers PEG and the zwitterion phosphorylcholine (PC). Recent evidence suggests that stiff hydrogels elicit increased immune cell adhesion to hydrogels, which we attempted to reduce by increasing hydrogel hydrophilicity. Surprisingly, hydrogels with the highest amount of zwitterionic comonomer elicited the highest FBR. Lowering the hydrogel modulus (165 to 3 kPa), or PC content (20 to 0 wt %), mitigated this effect. A high density of macrophages was found at the surface of implants associated with a high FBR, and mass spectrometry analysis of the proteins adsorbed to these gels implicated extracellular matrix, immune response, and cell adhesion protein categories as drivers of macrophage recruitment. Overall, we show that modulus regulates macrophage adhesion to zwitterionic-PEG hydrogels, and demonstrate that chemical modifications to hydrogels should be studied in parallel with their physical properties to optimize implant design.

  16. LysK CHAP endopeptidase domain is required for lysis of live staphylococcal cells.

    USDA-ARS?s Scientific Manuscript database

    LysK is a staphylococcal bacteriophage endolysin composed of three domains, an N-terminal cysteine, histidine-dependent amidohydrolases/peptidases (CHAP) endopeptidase domain (cleaves between D-alanine of the stem peptide and glycine of the cross-bridge peptide) a mid-protein amidase 2 domain (N-ace...

  17. Design and preparation of bi-functionalized short-chain modified zwitterionic nanoparticles.

    PubMed

    Hu, Fenglin; Chen, Kaimin; Xu, Hong; Gu, Hongchen

    2018-05-01

    An ideal nanomaterial for use in the bio-medical field should have a distinctive surface capable of effectively preventing nonspecific protein adsorption and identifying target bio-molecules. Recently, the short-chain zwitterion strategy has been suggested as a simple and novel approach to create outstanding anti-fouling surfaces. In this paper, the carboxyl end group of short-chain zwitterion-coated silica nanoparticles (SiO 2 -ZWS) was found to be difficult to functionalize via a conventional EDC/NHS strategy due to its rapid hydrolysis side-reactions. Hence, a series of bi-functionalized silica nanoparticles (SiO 2 -ZWS/COOH) were designed and prepared by controlling the molar ratio of 3-aminopropyltriethoxysilane (APTES) to short-chain zwitterionic organosiloxane (ZWS) in order to achieve above goal. The synthesized SiO 2 -ZWS/COOH had similar excellent anti-fouling properties compared with SiO 2 -ZWS, even in 50% fetal bovine serum characterized by DLS and turbidimetric titration. Subsequently, SiO 2 -ZWS/COOH 5/1 was chosen as a representative and then demonstrated higher detection signal intensity and more superior signal-to-noise ratios compare with the pure SiO 2 -COOH when they were used as a bio-carrier for chemiluminescence enzyme immunoassay (CLEIA). These unique bi-functionalized silica nanoparticles have many potential applications in the diagnostic and therapeutic fields. Reducing nonspecific protein adsorption and enhancing the immobilized efficiency of specific bio-probes are two of the most important issues for bio-carriers, particularly for a nanoparticle based bio-carrier. Herein, we designed and prepared a bi-functional nanoparticle with anti-fouling property and bio conjugation capacity for further bioassay by improving the short-chain zwitterionic modification strategy we have proposed previously. The heterogeneous surface of this nanoparticle showed effective anti-fouling properties both in model protein solutions and fetal bovine serum

  18. Zwitterionic modification of polyurethane membranes for enhancing the anti-fouling property.

    PubMed

    Liu, Peiming; Huang, Tao; Liu, Pingsheng; Shi, Shufeng; Chen, Qiang; Li, Li; Shen, Jian

    2016-10-15

    Polyurethane (PU) is a biopolymer that has been commonly used for biomedical applications. However, the biofouling phenomenon on the hydrophobic PU surface is one of the crucial issues that embarrassing its applications. Here, we report a facile & efficient approach to improve the anti-biofouling ability of the PU substrates. Active residues were firstly generated on the PU surface by using the low temperature air-plasma treatment, promoting the immobilization of the atom transfer radical polymerization (ATRP) initiators on the surface. Then, three types of zwitterionic polymer brushes, as well as PEG brushes, have been fabricated on the PU substrates through surface-initiated ATRP (SI-ATRP). Robust surface characterizations that capable of revealing the surface chemistry (including X-ray photoelectron spectroscopy (XPS) and wettability tests), and antifouling evaluations of the PU substrates (protein adsorption, platelet adhesion, and cell adhesion measurements) were performed. Results showed that three types of zwitterionic brushes have been successful grafted on the PU surface, respectively. And the three types of zwitterionic brushes, in general, significantly inhibited the protein adsorption, the platelet adhesion, and the cell adhesion on the PU surface, endowing a significantly improved anti-fouling ability to the PU substrates. Furthermore, we found that this facial zwitterionic surface modification did not compromise the mechanical property of the PU substrates. This strategy could be easily exploited to PU-based biomaterials to improve their performance in many applications. Copyright © 2016 Elsevier Inc. All rights reserved.

  19. Antifouling Thin-Film Composite Membranes by Controlled Architecture of Zwitterionic Polymer Brush Layer.

    PubMed

    Liu, Caihong; Lee, Jongho; Ma, Jun; Elimelech, Menachem

    2017-02-21

    In this study, we demonstrate a highly antifouling thin-film composite (TFC) membrane by grafting a zwitterionic polymer brush via atom-transfer radical-polymerization (ATRP), a controlled, environmentally benign chemical process. Initiator molecules for polymerization were immobilized on the membrane surface by bioinspired catechol chemistry, leading to the grafting of a dense zwitterionic polymer brush layer. Surface characterization revealed that the modified membrane exhibits reduced surface roughness, enhanced hydrophilicity, and lower surface charge. Chemical force microscopy demonstrated that the modified membrane displayed foulant-membrane interaction forces that were 1 order of magnitude smaller than those of the pristine TFC membrane. The excellent fouling resistance imparted by the zwitterionic brush layer was further demonstrated by significantly reduced adsorption of proteins and bacteria. In addition, forward osmosis fouling experiments with a feed solution containing a mixture of organic foulants (bovine-serum albumin, alginate, and natural organic matter) indicated that the modified membrane exhibited significantly lower water flux decline compared to the pristine TFC membrane. The controlled architecture of the zwitterionic polymer brush via ATRP has the potential for a facile antifouling modification of a wide range of water treatment membranes without compromising intrinsic transport properties.

  20. Identification of detergents for forensic fiber analysis.

    PubMed

    Heider, Emily C; Mujumdar, Nirvani; Campiglia, Andres D

    2016-11-01

    Trace fibers are an important form of trace evidence, and identification of exogenous substances on textile fibers provides valuable information about the origin of the fiber. Laundering textiles can provide a unique fluorescent spectral signature of the whitening agent in the detergent that adsorbs to the fiber. Using fluorescence microscopy, the spectral characteristics of seven detergents adsorbed to single fibers drawn from laundered textiles were investigated, and principal component analysis of clusters was used to characterize the type of detergent on the fiber. On dyed nylon fibers, spectra from eight different detergent pairs could be resolved and washed validation fibers correctly classified. On dyed acrylic fibers, five different detergent pairs could be resolved and identified. Identification of the detergent type may prove useful in matching a trace fiber to its bulk specimen of origin.

  1. Concentrated liquid detergent pod ingestion in children.

    PubMed

    Sidhu, Natasha; Jaeger, Matthew W

    2014-12-01

    Concentrated liquid detergent pods are an emerging public health hazard, especially in pediatric patients. Ingestion is a more common route of exposure for liquid detergent pods compared with non-pod detergents and it tends to be associated with more severe adverse effects. We present 3 cases that demonstrate the varied clinical symptoms resulting from detergent pod ingestion. These cases not only demonstrate findings such as gastrointestinal and respiratory symptoms but also show more rare neurological symptoms. The cases highlight the dangers of concentrated liquid detergent pod ingestion. To help prevent further life-threatening injuries, there is a need for more consumer information and provider knowledge about the potential adverse complications.

  2. Solubilization of glycoproteins of envelope viruses by detergents

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

    Berezin, V.E.; Zaides, V.M.; Artamsnov, A.F.

    1986-11-20

    The action of a number of known ionic and nonionic detergents, as well as the new nonionic detergent MESK, on envelope viruses was investigated. It was shown that the nonionic detergents MESK, Triton X-100, and octyl-..beta..-D-glucopyranoside selectively solubilize the outer glycoproteins of the virus particles. The nonionic detergent MESK has the mildest action. Using MESK, purified glycoproteins of influenza, parainfluenza, Venezuelan equine encephalomyelitis, vesicular stomatitis, rabies, and herpes viruses were obtained. The procedure for obtaining glycoproteins includes incubation of the virus suspension with the detergent MESK, removal of subvirus structures by centrifuging, and purification of glycoproteins from detergents by dialysis.more » Isolated glycoproteins retain a native structure and biological activity and possess high immunogenicity. The detergent MESK is promising for laboratory tests and with respect to the production of subunit vaccines.« less

  3. Zwitterion formation in titan ice analogs: reaction between HC3N and NH3.

    PubMed

    Couturier-Tamburelli, Isabelle; Sessouma, Bintou; Chiavassa, Thierry; Piétri, Nathalie

    2012-11-08

    A zwitterion is formed in the laboratory at low temperatures in the solid phase from the thermal reaction of HC(3)N and NH(3). We report for the first time its infrared spectrum. We study its reaction using Fourier transform infrared spectroscopy. Its reaction rate is estimated to be k(T) = 2.9 × 10(5) exp(-2.3 ± 0.1 (kJ mol(-1))/RT). Calculations using density functional theory (B3LYP/6-31g**) are used to characterize all the species (complexes, zwitterions, and transition states) and are in good agreement with the infrared spectra. The structure of the zwitterion is determined planar and it is characterized by a N-C bond around 1.5 Å.

  4. Detergent-compatible proteases: microbial production, properties, and stain removal analysis.

    PubMed

    Niyonzima, Francois Niyongabo; More, Sunil

    2015-01-01

    Proteases are one of the most important commercial enzymes used in various industrial domains such as detergent and leather industries. The alkaline proteases as well as other detergent-compatible enzymes such as lipases and amylases serve now as the key components in detergent formulations. They break down various stains during fabric washing. The search for detergent-compatible proteases with better properties is a continuous exercise. The current trend is to use detergent-compatible proteases that are stable over a wide temperature range. Although the proteases showing stability at elevated pH have the capacity to be used in detergent formulations, their usage can be significant if they are also stable and compatible with detergent and detergent ingredients, and also able to remove protein stains. Despite the existence of some reviews on alkaline proteases, there is no specification for the use of alkaline proteases as detergent additives. The present review describes the detergent-compatible proteases tested as detergent additives. An overview was provided for screening, optimization, purification, and properties of detergent compatible proteases, with an emphasis on the stability and compatibility of the alkaline proteases with the detergent and detergent compounds, as well as stain removal examination methods.

  5. Detergent-Specific Membrane Protein Crystallization Screens

    NASA Technical Reports Server (NTRS)

    Wiener, Michael

    2007-01-01

    A suite of reagents has been developed for three-dimensional crystallization of integral membranes present in solution as protein-detergent complexes (PDCs). The compositions of these reagents have been determined in part by proximity to the phase boundaries (lower consolute boundaries) of the detergents present in the PDCs. The acquisition of some of the requisite phase-boundary data and the preliminary design of several of the detergent- specific screens was supported by a NASA contract. At the time of expiration of the contract, a partial set of preliminary screens had been developed. This work has since been extended under non-NASA sponsorship, leading to near completion of a set of 20 to 30 different and unique detergent- specific 96-condition screens.

  6. Coproduction of detergent compatible bacterial enzymes and stain removal evaluation.

    PubMed

    Niyonzima, Francois N; More, Sunil S

    2015-10-01

    Most of the detergents that are presently produced contain the detergent compatible enzymes to improve and accelerate the washing performance by removing tough stains. The process is environment friendly as the use of enzymes in the detergent formulation reduces the utilization of toxic detergent constituents. The current trend is to use the detergent compatible enzymes that are active at low and ambient temperature in order to save energy and maintain fabric quality. As the detergent compatible bacterial enzymes are used together in the detergent formulation, it is important to co-produce the detergent enzymes in a single fermentation medium as the enzyme stability is assured, and production cost gets reduced enormously. The review reports on the production, purification, characterization and application of detergent compatible amylases, lipases, and proteases are available. However, there is no specific review or minireview on the concomitant production of detergent compatible amylases, lipases, and proteases. In this minireview, the coproduction of detergent compatible enzymes by bacterial species, enzyme stability towards detergents and detergent components, and stain release analysis were discussed. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. 40 CFR 80.163 - Detergent certification options.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 16 2010-07-01 2010-07-01 false Detergent certification options. 80... options. To be used to satisfy the detergency requirements under § 80.161(a), a detergent additive must be certified in accordance with the requirements of one or more of the options and suboptions described in this...

  8. Synthesis of mesoporous nano-hydroxyapatite by using zwitterions surfactant

    EPA Science Inventory

    Mesoporous nano-hydroxyapatite (mn-HAP) was successfully synthesized via a novel micelle-templating method using lauryl dimethylaminoacetic acid as zwitterionic surfactant. The systematic use of such a surfactant in combination with microwave energy inputenables the precise contr...

  9. Physiological stability and renal clearance of ultrasmall zwitterionic gold nanoparticles: Ligand length matters

    NASA Astrophysics Data System (ADS)

    Ning, Xuhui; Peng, Chuanqi; Li, Eric S.; Xu, Jing; Vinluan, Rodrigo D.; Yu, Mengxiao; Zheng, Jie

    2017-05-01

    Efficient renal clearance has been observed from ultrasmall zwitterionic glutathione-coated gold nanoparticles (GS-AuNPs), which have broad preclinical applications in cancer diagnosis and kidney functional imaging. However, origin of such efficient renal clearance is still not clear. Herein, we conducted head-to-head comparison on physiological stability and renal clearance of two zwitterionic luminescent AuNPs coated with cysteine and glycine-cysteine (Cys-AuNPs and Gly-Cys-AuNPs), respectively. While both of them exhibited similar surface charges and the same core sizes, additional glycine slightly increased the hydrodynamic diameter of the AuNPs by 0.4 nm but significantly enhanced physiological stability of the AuNPs as well as altered their clearance pathways. These studies indicate that the ligand length, in addition to surface charges and size, also plays a key role in the physiological stability and renal clearance of ultrasmall zwitterionic inorganic NPs.

  10. Pediatric exposure to laundry detergent pods.

    PubMed

    Valdez, Amanda L; Casavant, Marcel J; Spiller, Henry A; Chounthirath, Thiphalak; Xiang, Huiyun; Smith, Gary A

    2014-12-01

    Laundry detergent pods are a new product in the US marketplace. This study investigates the epidemiologic characteristics and outcomes of laundry detergent pod exposures among young children in the United States. Using data from the National Poison Data System, exposures to laundry detergent pods among children younger than 6 years of age during 2012-2013 were investigated. There were 17 230 children younger than 6 years exposed to laundry detergent pods in 2012-2013. From March 2012 to April 2013, the monthly number of exposures increased by 645.3%, followed by a 25.1% decrease from April to December 2013. Children younger than 3 years accounted for 73.5% of cases. The major route of exposure was ingestion, accounting for 79.7% of cases. Among exposed children, 4.4% were hospitalized and 7.5% experienced a moderate or major medical outcome. A spectrum of clinical effects from minor to serious was seen with ingestion and ocular exposures. There were 102 patients (0.6%) exposed to a detergent pod via ingestion, aspiration, or a combination of routes, including ingestion, who required tracheal intubation. There was 1 confirmed death. Laundry detergent pods pose a serious poisoning risk to young children. This nationwide study underscores the need for increased efforts to prevent exposure of young children to these products, which may include improvements in product packaging and labeling, development of a voluntary product safety standard, and public education. Product constituent reformulation is another potential strategy to mitigate the severity of clinical effects of laundry detergent pod exposure. Copyright © 2014 by the American Academy of Pediatrics.

  11. Experimental Analysis of Detergency Phenomena and Investigation of a Next-generation Detergency System.

    PubMed

    Gotoh, Keiko

    2017-01-01

    The detergency of products, mainly textiles, was evaluated using various experimental systems and discussed from the viewpoint of interfacial phenomena. The detergency phenomena observed for geometrically simple model systems were explained in terms of the total potential energy of interaction between the soil and the substrate, which was calculated as the sum of the electrical double layer, Lifshitzvan der Waals, and acid-base interactions using electrokinetic potentials and surface free energy components. Cleaning experiments using artificially soiled fabrics were performed using electro-osmotic flow and ultrasound as mechanical actions for soil removal, and the results were compared with those obtained with mechanical actions commonly used in textile washing. Simultaneous hydrophilization of the substrate and soil by an atmospheric pressure plasma jet remarkably improved the detergency in aqueous solutions. The application of the atmospheric pressure plasma jet to anti-fouling textiles was also proposed.

  12. Improved detergent-based recovery of polyhydroxyalkanoates (PHAs).

    PubMed

    Yang, Yung-Hun; Brigham, Christopher; Willis, Laura; Rha, ChoKyun; Sinskey, Anthony

    2011-05-01

    Extracting polyhydroxyalkanoate (PHA) polymer from bacterial cells often involves harsh conditions, including use of environmentally harmful solvents. We evaluated different detergents under various conditions to extract PHA from Ralstonia eutropha and Escherichia coli cells. Most detergents tested recovered highly pure PHA polymer from cells in amounts that depended on the percentage of polymer present in the cell. Detergents such as linear alkylbenzene sulfonic acid (LAS-99) produced a high yield of high purity polymer, and less detergent was needed compared to the amount of SDS to produce comparable yields. LAS-99 also has the advantage of being biodegradable and environmentally safe. Chemical extraction of PHA with detergents could potentially minimize or eliminate the need to use harsh organic solvents, thus making industrial PHA production a cleaner technology process. © Springer Science+Business Media B.V. 2011

  13. Designing Mixed Detergent Micelles for Uniform Neutron Contrast

    DOE PAGES

    Oliver, Ryan C.; Pingali, Sai Venkatesh; Urban, Volker S.

    2017-09-29

    Micelle-forming detergents provide an amphipathic environment that mimics lipid bilayers and are important tools used to solubilize and stabilize membrane proteins in solution for in vitro structural investigations. Small-angle neutron scattering (SANS) performed at the neutron contrast match point of detergent molecules allows observing the scattering signal from membrane proteins unobstructed by contributions from the detergent. However, we show here that even for a perfectly average-contrast matched detergent there arises significant core-shell scattering from the contrast difference between aliphatic detergent tails and hydrophilic head groups. This residual signal at the average detergent contrast match point interferes with interpreting structural datamore » of membrane proteins. This complication is often made worse by the presence of excess empty (protein-free) micelles. Here, we present an approach for the rational design of mixed micelles containing a deuterated detergent analog, which eliminates neutron contrast between core and shell, and allows the micelle scattering to be fully contrast matched to unambiguously resolve membrane protein structure using solution SANS.« less

  14. Designing Mixed Detergent Micelles for Uniform Neutron Contrast

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

    Oliver, Ryan C.; Pingali, Sai Venkatesh; Urban, Volker S.

    Micelle-forming detergents provide an amphipathic environment that mimics lipid bilayers and are important tools used to solubilize and stabilize membrane proteins in solution for in vitro structural investigations. Small-angle neutron scattering (SANS) performed at the neutron contrast match point of detergent molecules allows observing the scattering signal from membrane proteins unobstructed by contributions from the detergent. However, we show here that even for a perfectly average-contrast matched detergent there arises significant core-shell scattering from the contrast difference between aliphatic detergent tails and hydrophilic head groups. This residual signal at the average detergent contrast match point interferes with interpreting structural datamore » of membrane proteins. This complication is often made worse by the presence of excess empty (protein-free) micelles. Here, we present an approach for the rational design of mixed micelles containing a deuterated detergent analog, which eliminates neutron contrast between core and shell, and allows the micelle scattering to be fully contrast matched to unambiguously resolve membrane protein structure using solution SANS.« less

  15. Superior Antifouling Performance of a Zwitterionic Peptide Compared to an Amphiphilic, Non-Ionic Peptide.

    PubMed

    Ye, Huijun; Wang, Libing; Huang, Renliang; Su, Rongxin; Liu, Boshi; Qi, Wei; He, Zhimin

    2015-10-14

    The aim of this study was to explore the influence of amphiphilic and zwitterionic structures on the resistance of protein adsorption to peptide self-assembled monolayers (SAMs) and gain insight into the associated antifouling mechanism. Two kinds of cysteine-terminated heptapeptides were studied. One peptide had alternating hydrophobic and hydrophilic residues with an amphiphilic sequence of CYSYSYS. The other peptide (CRERERE) was zwitterionic. Both peptides were covalently attached onto gold substrates via gold-thiol bond formation. Surface plasmon resonance analysis results showed that both peptide SAMs had ultralow or low protein adsorption amounts of 1.97-11.78 ng/cm2 in the presence of single proteins. The zwitterionic peptide showed relatively higher antifouling ability with single proteins and natural complex protein media. We performed molecular dynamics simulations to understand their respective antifouling behaviors. The results indicated that strong surface hydration of peptide SAMs contributes to fouling resistance by impeding interactions with proteins. Compared to the CYSYSYS peptide, more water molecules were predicted to form hydrogen-bonding interactions with the zwitterionic CRERERE peptide, which is in agreement with the antifouling test results. These findings reveal a clear relation between peptide structures and resistance to protein adsorption, facilitating the development of novel peptide-containing antifouling materials.

  16. Developmental assessment of the Fort St. Vrain version of the Composite HTGR Analysis Program (CHAP-2)

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

    Stroh, K.R.

    1980-01-01

    The Composite HTGR Analysis Program (CHAP) consists of a model-independent systems analysis mainframe named LASAN and model-dependent linked code modules, each representing a component, subsystem, or phenomenon of an HTGR plant. The Fort St. Vrain (FSV) version (CHAP-2) includes 21 coded modules that model the neutron kinetics and thermal response of the core; the thermal-hydraulics of the reactor primary coolant system, secondary steam supply system, and balance-of-plant; the actions of the control system and plant protection system; the response of the reactor building; and the relative hazard resulting from fuel particle failure. FSV steady-state and transient plant data are beingmore » used to partially verify the component modeling and dynamic smulation techniques used to predict plant response to postulated accident sequences.« less

  17. 40 CFR 80.141 - Interim detergent gasoline program.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ...) “Carburetor Cleanliness Test Procedure, State-of-the-Art Summary, Report: 1973-1981”, Coordinating Research... ultimate consumer; (ii) All additized post-refinery component (PRC); and (iii) All detergent additives sold... who manufacture, supply, or transfer detergent additives or detergent-additized post-refinery...

  18. Crystal structure of zwitterionic bisimidazolium sulfonates

    NASA Astrophysics Data System (ADS)

    Kohmoto, Shigeo; Okuyama, Shinpei; Yokota, Nobuyuki; Takahashi, Masahiro; Kishikawa, Keiki; Masu, Hyuma; Azumaya, Isao

    2012-05-01

    Crystal structures of three zwitterionic bisimidazolium salts 1-3 in which imidazolium sulfonate moieties were connected with aromatic linkers, p-xylylene, 4,4'-dimethylenebiphenyl, and phenylene, respectively, were examined. The latter two were obtained as hydrates. An S-shaped molecular structure in which the sulfonate moiety was placed on the imidazolium ring was observed for 1. A helical array of hydrated water molecules was obtained for 2 while a linear array of hydrated water molecules was observed for 3.

  19. Purification and characterization of a membrane-associated 3,3',5-triiodo-L-thyronine binding protein from a human carcinoma cell line.

    PubMed Central

    Cheng, S Y; Hasumura, S; Willingham, M C; Pastan, I

    1986-01-01

    A membrane-associated binding protein for 3,3',5-triiodo-L-thyronine (T3) was purified to apparent homogeneity from A431 human epidermoid carcinoma cells. A431 cells were specifically labeled with the N-bromoacetyl derivative of T3 labeled with 125I at the 3' position (BrAc[125I]T3) and were extracted with 3-[3-(cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), a zwitterionic detergent. The solubilized BrAc[125I]T3-labeled protein was successively purified by chromatography on Sephadex G-200 and QAE-Sephadex followed by NaDod-SO4/PAGE. Approximately 0.2 mg of purified protein was obtained from 2.5 X 10(9) cells, which represents a 3000-fold purification. The membrane-associated T3 binding protein is an acidic protein with a pI of 5.1 and an apparent molecular mass of 55,000 daltons determined by NaDodSO4/PAGE. Polyclonal antibodies against the 55-kDa protein were prepared and used in indirect immunofluorescence to show that the 55-kDa protein was mainly found in the nuclear envelope and endoplasmic reticulum. Images PMID:3006034

  20. Movement and fate of detergents in groundwater: a field study

    USGS Publications Warehouse

    Thurman, E.M.; Barber, L.B.; LeBlanc, D.

    1986-01-01

    The major cations, anions, and detergents in a plume of contaminated groundwater at Otis Air Base on Cape Cod (Mass., U.S.A.) have moved approximately 3.5 km down gradient from the disposal beds. We hypothesize that the detergents form two distinct plumes, which consist of alkyl benzene sulfonates (ABS) detergents and linear alkyl sulfonates (LAS) and sodium dodecyl sulfate (NaLS) detergents. The ABS detergents were deposited from approximately 1940 through 1965, when ABS detergents were banned. From 1965 to the present, LAS and NaLS detergents were in the sewage. The ABS detergents appear to be transported in the aquifer at the same rate as the specific conductance (major cations and anions) and boron, which are currently used as conservative tracers of the plume of contaminated groundwater. There appears to be little or no biological degradation of the ABS detergents in the aquifer, based on their concentration in the plume. On the other hand, the LAS and NaLS detergents have degraded rapidly and have been detected only 0.6 km down gradient. The roleof the detergents in the transport of other organic compounds in the plume is nuclear. There is a separation of the ABS detergent plume and the volatile organic compound plume; however, the time of entry of the detergents and the volatile organic compounds is unknown. Therefore, it is not possible to conclude on the interaction of these two classes of compounds. ?? 1986.

  1. The self-assembling zwitterionic form of L-phenylalanine at neutral pH.

    PubMed

    Mossou, Estelle; Teixeira, Susana C M; Mitchell, Edward P; Mason, Sax A; Adler-Abramovich, Lihi; Gazit, Ehud; Forsyth, V Trevor

    2014-03-01

    The title zwitterion (2S)-2-azaniumyl-1-hydroxy-3-phenylpropan-1-olate, C9H11NO2, also known as L-phenylalanine, was characterized using synchrotron X-rays. It crystallized in the monoclinic space group P21 with four molecules in the asymmetric unit. The 0.62 Å resolution structure is assumed to be closely related to the fibrillar form of phenylalanine, as observed by electron microscopy and electron diffraction. The structure exists in a zwitterionic form in which π-π stacking and hydrogen-bonding interactions are believed to form the basis of the self-assembling properties.

  2. Soaps and Detergents--A 'Social' Treatment.

    ERIC Educational Resources Information Center

    Rust, S. C.

    1979-01-01

    Describes how social aspects of science can be incorporated into teaching soap and detergents in British secondary chemistry and general science courses. Historical background to the use and production of soap and the development of detergents are also presented. (HM)

  3. Detergents as selective inhibitors and inactivators of enzymes.

    PubMed

    Vincenzini, M T; Favilli, F; Stio, M; Vanni, P; Treves, C

    1985-01-01

    In order to study the detergent-enzyme interaction and to clarify whether such an interaction produces specific or non-specific effects, we investigated the action of natural and synthetic detergents on enzymatic systems of different levels of complexity (crystalline enzymes, crude homogenates, organ preparations, organisms in toto i.e. rats and germinating seeds). The enzyme-detergent interaction was examined both as a time-independent phenomenon (inhibition) and as a time-dependent phenomenon (inactivation). In in vitro experiments a clear inhibition of pyridine-dependent dehydrogenases by long-chain anionic detergents was found. Cationic detergents have their greatest effect on lipase, LDH, MDH and ICDH from rat liver homogenates. At low concentrations SDS inactivates all the dehydrogenase enzymes studied. With high concentrations (10 mM) of SDS and dodecyltrimethylammonium bromide (C12), there was a sharp and non-specific decrease of enzymatic activities. In the in vivo studies, rats were given detergents to drink; the cationic detergent (C12) was far more effective than SDS with enzymes from both intestine and liver homogenates. SDS and C12 do not seem to interfere with enzyme activities at the beginning of the germination of Pinus pinea and Triticum durum seeds. However a marked reduction of activities does occur at the respective maximum germination times of these seeds. The nonionic detergent is ineffective both as inhibitor and as inactivator.

  4. Comparative studies on ecotoxicology of synthetic detergents.

    PubMed

    Lal, H; Misra, V; Viswanathan, P N; Krishna Murti, C R

    1983-12-01

    To predict the comparative toxicological response of synthetic detergents on aquatic ecosystems, the effects of various concentrations of neutralized alkyl benzene sulfonate were studied. The median tolerance limit at 48 hr, 95% confidence limit, slope function, presumable harmless concentration, and rate of survival of different species of aquatic fauna such as water fleas (Daphnia magna), mosquito larvae (Culex pipiens), slug worms (Tubifex rivulorum), snails (Lymnaea vulgaris), tadpoles (Rana cyanophlyctis), and fish fingerlings (Cirrhina mrigala) were followed at 0, 24, 48, 72, and 96 hr. Any effect on quality of the water was also tested after the addition of various concentrations of detergents. The results showed that water fleas are more susceptible to detergent toxicity than fish fingerlings, tadpoles, slug worms, snails, and mosquito larvae. Behavioral changes were also observed as an index for detergent toxicity. The relative toxicity of the detergents to various species is discussed in relation to selective ecotoxicological response.

  5. Zwitterionic Antifouling Coatings for the Purification of High-Salinity Shale Gas Produced Water.

    PubMed

    Yang, Rong; Goktekin, Esma; Gleason, Karen K

    2015-11-03

    Fouling refers to the undesirable attachment of organic molecules and microorganisms to submerged surfaces. It is an obstacle to the purification of shale gas produced water and is currently without an effective solution due to the highly contaminated nature of produced water. Here, we demonstrate the direct vapor application of a robust zwitterionic coating to a variety of substrates. The coating remains unprecedentedly hydrophilic, smooth, and effectively antifouling in extremely high salinity solutions (with salt concentration of 200,000 ppm). The fouling resistance is assessed rapidly and quantitatively with a molecular force spectroscopy-based method and corroborated using quartz crystal microbalance system with dissipation monitoring. Grazing angle attenuated total reflectance Fourier transform infrared is used in combination with X-ray photoelectron spectroscopy, atomic force microscope, and in situ spectroscopic ellipsometry to lend insight into the underlying mechanism for the exceptional stability and effectiveness of the zwitterionic coating under high-salinity conditions. A unique coating architecture, where the surface is concentrated with mobile zwitterionic moieties while the bulk is cross-linked to enhance coating durability, was discovered to be the origin of its stable fouling resistance under high salinity. Combined with previously reported exceptional stability in highly oxidative environments and strong fouling resistance to oil and grease, the zwitterionic surface here has the potential to enable low-cost, membrane-based techniques for the purification of produced water and to eventually balance the favorable economics and the concerning environmental impacts of the hydraulic fracturing industry.

  6. Intermolecular detergent-membrane protein noes for the characterization of the dynamics of membrane protein-detergent complexes.

    PubMed

    Eichmann, Cédric; Orts, Julien; Tzitzilonis, Christos; Vögeli, Beat; Smrt, Sean; Lorieau, Justin; Riek, Roland

    2014-12-11

    The interaction between membrane proteins and lipids or lipid mimetics such as detergents is key for the three-dimensional structure and dynamics of membrane proteins. In NMR-based structural studies of membrane proteins, qualitative analysis of intermolecular nuclear Overhauser enhancements (NOEs) or paramagnetic resonance enhancement are used in general to identify the transmembrane segments of a membrane protein. Here, we employed a quantitative characterization of intermolecular NOEs between (1)H of the detergent and (1)H(N) of (2)H-perdeuterated, (15)N-labeled α-helical membrane protein-detergent complexes following the exact NOE (eNOE) approach. Structural considerations suggest that these intermolecular NOEs should show a helical-wheel-type behavior along a transmembrane helix or a membrane-attached helix within a membrane protein as experimentally demonstrated for the complete influenza hemagglutinin fusion domain HAfp23. The partial absence of such a NOE pattern along the amino acid sequence as shown for a truncated variant of HAfp23 and for the Escherichia coli inner membrane protein YidH indicates the presence of large tertiary structure fluctuations such as an opening between helices or the presence of large rotational dynamics of the helices. Detergent-protein NOEs thus appear to be a straightforward probe for a qualitative characterization of structural and dynamical properties of membrane proteins embedded in detergent micelles.

  7. Concentrating membrane proteins using ultrafiltration without concentrating detergents.

    PubMed

    Feroz, Hasin; Vandervelden, Craig; Ikwuagwu, Bon; Ferlez, Bryan; Baker, Carol S; Lugar, Daniel J; Grzelakowski, Mariusz; Golbeck, John H; Zydney, Andrew L; Kumar, Manish

    2016-10-01

    Membrane proteins (MPs) are of rapidly growing interest in the design of pharmaceutical products, novel sensors, and synthetic membranes. Ultrafiltration (UF) using commercially available centrifugal concentrators is typically employed for laboratory-scale concentration of low-yield MPs, but its use is accompanied by a concomitant increase in concentration of detergent micelles. We present a detailed analysis of the hydrodynamic processes that control detergent passage during ultrafiltration of MPs and propose methods to optimize detergent passage during protein concentration in larger-scale membrane processes. Experiments were conducted using nonionic detergents, octyl-β-D glucoside (OG), and decyl-β-D maltoside (DM) with the bacterial water channel protein, Aquaporin Z (AqpZ) and the light driven chloride pump, halorhodopsin (HR), respectively. The observed sieving coefficient (So ), a measure of detergent passage, was evaluated in both stirred cell and centrifugal systems. So for DM and OG increased with increasing filtrate flux and decreasing shear rates in the stirred cell, that is, with increasing concentration polarization (CP). Similar effects were observed during filtration of MP-detergent (MPD) micelles. However, lower transmission was observed in the centrifugal system for both detergent and MPD systems. This is attributed to free convection-induced shear and hence reduced CP along the membrane surface during centrifugal UF. Thus to concentrate MPs without retention of detergent, design of UF systems that promote CP is required. Biotechnol. Bioeng. 2016;113: 2122-2130. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  8. Greatly decreased susceptibility of nonmetabolizing cells towards detergents.

    PubMed Central

    Komor, E; Weber, H; Tanner, W

    1979-01-01

    The addition of different detergents to Chlorella cells that had previously accumulated 6-deoxyglucose causes a rapid release of the hexose analogue into the medium. This effect is independent of the nature of the detergent and is observed only when the cells are in an energized state. Thus, in the presence of the uncoupler p-trifluoromethoxycarbonylcyanide phenylhydrazone or of inhibitors such as N-ethylmaleimide, the cells show a greatly reduced susceptibility towards detergents. Similarly, the detergent-induced loss of accumulated alpha-aminoisobutyric acid from Saccharomyces cerevisiae and of potassium from Escherichia coli is also strongly affected by the energy state of the cells. The differential susceptibility of energized and nonenergized cells was observed at all detergent concentrations tested. Measurements of substrate efflux at different concentrations of Triton indicated that only Triton monomers are responsible for the increase in permeability. The absorption of [14C]Triton X-100 by Chlorella and the binding of detergent to the cells were measured in the presence of metabolic inhibitors. Again, nonenergized cells bound a significantly lower amount of Triton X-100. The amphiphilic antibiotic nystatin produced effects on cell permeability similar to those of detergents, whereas toluene, which is apolar, gave opposite results. PMID:377284

  9. 40 CFR 80.161 - Detergent additive certification program.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... requirements. For a detergent additive package to be certified as eligible for use by detergent blenders in...: (i) The compositional data required under § 79.21(a) of this chapter shall include the information... chapter shall be reported to EPA in units of gallons of detergent additive package per 1000 gallons of...

  10. 40 CFR 80.161 - Detergent additive certification program.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... requirements. For a detergent additive package to be certified as eligible for use by detergent blenders in...: (i) The compositional data required under § 79.21(a) of this chapter shall include the information... chapter shall be reported to EPA in units of gallons of detergent additive package per 1000 gallons of...

  11. 40 CFR 80.161 - Detergent additive certification program.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... requirements. For a detergent additive package to be certified as eligible for use by detergent blenders in...: (i) The compositional data required under § 79.21(a) of this chapter shall include the information... chapter shall be reported to EPA in units of gallons of detergent additive package per 1000 gallons of...

  12. Detergent Screening and Purification of the Human Liver ABC Transporters BSEP (ABCB11) and MDR3 (ABCB4) Expressed in the Yeast Pichia pastoris

    PubMed Central

    Stindt, Jan; Smits, Sander H. J.; Schmitt, Lutz

    2013-01-01

    The human liver ATP-binding cassette (ABC) transporters bile salt export pump (BSEP/ABCB11) and the multidrug resistance protein 3 (MDR3/ABCB4) fulfill the translocation of bile salts and phosphatidylcholine across the apical membrane of hepatocytes. In concert with ABCG5/G8, these two transporters are responsible for the formation of bile and mutations within these transporters can lead to severe hereditary diseases. In this study, we report the heterologous overexpression and purification of human BSEP and MDR3 as well as the expression of the corresponding C-terminal GFP-fusion proteins in the yeast Pichia pastoris. Confocal laser scanning microscopy revealed that BSEP-GFP and MDR3-GFP are localized in the plasma membrane of P. pastoris. Furthermore, we demonstrate the first purification of human BSEP and MDR3 yielding ∼1 mg and ∼6 mg per 100 g of wet cell weight, respectively. By screening over 100 detergents using a dot blot technique, we found that only zwitterionic, lipid-like detergents such as Fos-cholines or Cyclofos were able to extract both transporters in sufficient amounts for subsequent functional analysis. For MDR3, fluorescence-detection size exclusion chromatography (FSEC) screens revealed that increasing the acyl chain length of Fos-Cholines improved monodispersity. BSEP purified in n-dodecyl-β-D-maltoside or Cymal-5 after solubilization with Fos-choline 16 from P. pastoris membranes showed binding to ATP-agarose. Furthermore, detergent-solubilized and purified MDR3 showed a substrate-inducible ATPase activity upon addition of phosphatidylcholine lipids. These results form the basis for further biochemical analysis of human BSEP and MDR3 to elucidate the function of these clinically relevant ABC transporters. PMID:23593265

  13. Modification of silicone elastomer with zwitterionic silane for durable antifouling properties.

    PubMed

    Yeh, Shiou-Bang; Chen, Chien-Sheng; Chen, Wen-Yih; Huang, Chun-Jen

    2014-09-30

    Biofouling on medical devices generally causes adverse complications, such as thrombosis, infection, and pathogenic calcification. Silicone is a widely used material for medical applications. Its surface modification typically encounters undesirable "hydrophobic recovery", leading to deterioration of surface engineering. In this study, we developed a stable superhydrophilic zwitterionic interface on polydimethylsiloxane (PDMS) elastomer by covalent silanization of sulfobetaine silane (SBSi) to resist nonspecific adsorption of bacteria, proteins, and lipids. SBSi is a zwitterionic organosilane assembly, enabling resisting surface reconstruction by forming a cross-linked network and polar segregation. Surface elemental composition was confirmed by X-ray photoelectron spectroscopy (XPS), and the long-term stability of modification was accessed using a contact angle goniometer. The biofouling tests were carried out by exposing substrates to bacterial, protein, and lipid solutions, revealing the excellent bioinertness of SBSi-tailored PDMS, even after 30 day storage in ambient. For the real-world application, we modified commercially available silicone hydrogel contact lenses with developed zwitterionic silane, presenting its antibacterial adhesion property. Moreover, the cytotoxicity of SBSi was accessed with NIH-3T3 fibroblast by the MTT assay, showing negligible cytotoxicity up to a concentration of 5 mM. Consequently, the strategy of surface engineering in this work can effectively retard the "hydrophobic recovery" occurrence and can be applied to other silicone-based medical devices in a facile way.

  14. Dissociation, aggregation of sesame alpha-globulin in nonionic detergent solution.

    PubMed

    Lakshmi, T S; Nandi, P K

    1978-10-01

    Nonionic detergents Triton X-100 and Brij 36T induce dissociation and aggregation of the protein sesame alpha-globulin above the critical micelle concentrations (cmc) of the detergents. Spectrophotometric titration in Triton shows no change in the pKInt value of the tyrosyl groups at 1x10-3 M detergent where both dissociation and aggregation of the protein are observed. Fluorescence measurement does not indicate any change in the environment of the tryptophan groups of the protein in Brij. Viscosity measurements show no major conformational change of the protein in the detergent solution. Binding measurements suggest that perhaps micelles of the detergent predominantly bind to the protein. The detergent micelles preferentially bind to the exposed hydrophobic surfaces of the protein subunits. The association of the protein detergent complex through electrostatic interaction is probably responsible for the formation of the aggregates.

  15. Efficient CO2 capture by tertiary amine-functionalized ionic liquids through Li+-stabilized zwitterionic adduct formation

    PubMed Central

    Yang, Zhen-Zhen

    2014-01-01

    Summary Highly efficient CO2 absorption was realized through formation of zwitterionic adducts, combining synthetic strategies to ionic liquids (ILs) and coordination. The essence of our strategy is to make use of multidentate cation coordination between Li+ and an organic base. Also PEG-functionalized organic bases were employed to enhance the CO2-philicity. The ILs were reacted with CO2 to form the zwitterionic adduct. Coordination effects between various lithium salts and neutral ligands, as well as the CO2 capacity of the chelated ILs obtained were investigated. For example, the CO2 capacity of PEG150MeBu2N increased steadily from 0.10 to 0.66 (mol CO2 absorbed per mol of base) through the formation of zwitterionic adducts being stabilized by Li+. PMID:25246955

  16. Synthesis of zwitterionic salts of pyridinium-Meldrum acid and barbiturate through unique four-component reactions.

    PubMed

    Wang, Qi-Fang; Hui, Li; Hou, Hong; Yan, Chao-Guo

    2010-03-08

    An efficient synthetic procedure for the preparation of the unusual charge-separated pyridinium-Meldrum acid and N,N-dimethylbarbiturate acid zwitterionic salts was developed though a unique one-pot four-component reaction involving pyridine, aromatic aldehyde, Meldrum acid or N,N-dimethylbarbituric acid, and p-nitrobenzyl bromide in acetonitrile. By varying combinations of four components involving nitrogen-containing heterocycles, we conveniently established reactive alpha-halomethylene compounds, aldehydes and beta-dicarbonyl compounds a library of zwitterionic salts.

  17. Characterization of the protease activity of detergents: laboratory practicals for studying the protease profile and activity of various commercial detergents.

    PubMed

    Valls, Cristina; Pujadas, Gerard; Garcia-Vallve, Santi; Mulero, Miquel

    2011-07-01

    Detergent enzymes account for about 30% of the total worldwide production of enzymes and are one of the largest and most successful applications of modern industrial biotechnology. Proteases can improve the wash performance of household, industrial, and institutional laundry detergents used to remove protein-based stains such as blood, grass, body fluids, and food soils. This article describes two easy and cheap laboratory exercises to study the presence, profile, and basic enzymology of detergent proteases. These laboratory practicals are based on the determination of the detergent protease activity of various commercial detergents using the N-succinyl-L-alanyl-L-alanyl-L-prolyl-L-phenylalanine p-nitroanilide method and the bovine serum albumin degradation capacity. Students are also required to elucidate the enzymatic subtype of detergent proteases by studying the inhibitory potential of several types of protease inhibitors revealed by the same experimental methodology. Additionally, the results of the exercises can be used to provide additional insights on elementary enzymology by studying the influence of several important parameters on protease activity such as temperature (in this article) and the influence of pH and effects of surfactants and oxidizers (proposed). Students also develop laboratory skills, problem-solving capacities, and the ability to write a laboratory report. The exercises are mainly designed for an advanced undergraduate project in the biochemistry and biotechnology sciences. Globally, these laboratory practicals show students the biotechnological applications of proteases in the detergent industry and also reinforce important enzymology concepts. Copyright © 2010 Wiley Periodicals, Inc.

  18. Advances in protease engineering for laundry detergents.

    PubMed

    Vojcic, Ljubica; Pitzler, Christian; Körfer, Georgette; Jakob, Felix; Ronny Martinez; Maurer, Karl-Heinz; Schwaneberg, Ulrich

    2015-12-25

    Proteases are essential ingredients in modern laundry detergents. Over the past 30 years, subtilisin proteases employed in the laundry detergent industry have been engineered by directed evolution and rational design to tailor their properties towards industrial demands. This comprehensive review discusses recent success stories in subtilisin protease engineering. Advances in protease engineering for laundry detergents comprise simultaneous improvement of thermal resistance and activity at low temperatures, a rational strategy to modulate pH profiles, and a general hypothesis for how to increase promiscuous activity towards the production of peroxycarboxylic acids as mild bleaching agents. The three protease engineering campaigns presented provide in-depth analysis of protease properties and have identified principles that can be applied to improve or generate enzyme variants for industrial applications beyond laundry detergents. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Detergent-Fearing Milk.

    ERIC Educational Resources Information Center

    Hill, Diane

    1997-01-01

    Describes an activity that demonstrates among the following: diffusion; cohesion and adhesion; properties of surface tension which include wicking, hydrophilic, and hydrophobic molecular behaviors; and break up of fat clusters by liquid dishwashing detergent. (DDR)

  20. Is chronic detergent ingestion harmful to the gut?

    PubMed Central

    Mercurius-Taylor, L A; Jayaraj, A P; Clark, C G

    1984-01-01

    Synthetic detergents are used in large quantities as household and industrial cleaners. Because of the common practice of leaving dishes washed in detergent solutions to dry without rinsing these compounds are ingested. We have calculated that an adult takes in about 1 mg/kg detergent a day and babies can be administered between seven and 10 mg/kg a day. Rats were fed a dose of 100 mg/kg a day in a pilot experiment and gross abnormalities were found in the gastrointestinal tract, the most striking being subtotal villous atrophy of the small bowel mucosa and glandular atrophy in the colon. These changes were not reversible 12 weeks after cessation of detergent administration. Images PMID:6722057

  1. Biodegradability, toxicity and mutagenicity of detergents: Integrated experimental evaluations.

    PubMed

    Pedrazzani, Roberta; Ceretti, Elisabetta; Zerbini, Ilaria; Casale, Rosario; Gozio, Eleonora; Bertanza, Giorgio; Gelatti, Umberto; Donato, Francesco; Feretti, Donatella

    2012-10-01

    The widespread use of detergents has raised concern with regard to the environmental pollution caused by their active ingredients, which are biorefractory, toxic and persistent. Since detergents are complex mixtures of different substances, in which synergistic effects may occur, we aimed to assess the mutagenicity of different detergent formulations, taking into account aquatic toxicity and ready biodegradability. We performed a ready biodegradability test (OECD 301 F), Daphnia magna and Vibrio fischeri toxicity tests, and mutagenicity tests (Salmonella/microsome test, Allium cepa test and comet assay). Six detergent formulations were examined, 3 pre-manufacture and 3 commercially available. All detergents presented ready biodegradability. EC50 values varied for all products, according to the marker organism used, but were always higher than the more stringent value considered for aquatic toxicity assessment (V. fischeri 10-60 mg/L; D. magna 25-300 mg/L; A. cepa 250-2000 mg/L). None of the detergents caused mutations in bacteria. However, one commercial ecolabelled product induced an increase in micronucleus frequency in A. cepa root cells. All pre-manufacture detergents and one commercial one, which gave negative results in the Ames and A. cepa tests, induced DNA damage in human leukocytes. A more accurate evaluation of the environmental impact of complex mixtures such as detergents requires a battery of tests to describe degradation, as well as toxicological and mutagenic features. Copyright © 2012 Elsevier Inc. All rights reserved.

  2. Correlative microscopy of detergent granules.

    PubMed

    van Dalen, G; Nootenboom, P; Heussen, P C M

    2011-03-01

    The microstructure of detergent products for textile cleaning determines to a large extent the physical properties of these products. Correlative microscopy was used to reveal the microstructure by reconciling images obtained by scanning electron microscopy with energy dispersive X-ray analysis, X-ray microtomography and Fourier transform infrared microscopy. These techniques were applied on the same location of a subsample of a spray-dried detergent base powder embedded in polyacrylate. In this way, the three-dimensional internal and external structure of detergent granules could be investigated from milli to nano scale with detailed spatial information about the components present. This will generate knowledge how to design optimal microstructures for laundry products to obtain product properties demanded by the market. This method is also very useful for other powder systems used in a large variety of industries (e.g. for pharmaceutical, food, ceramic and metal industries). © 2010 The Authors Journal of Microscopy © 2010 The Royal Microscopical Society.

  3. hERG blocking potential of acids and zwitterions characterized by three thresholds for acidity, size and reactivity.

    PubMed

    Nikolov, Nikolai G; Dybdahl, Marianne; Jónsdóttir, Svava Ó; Wedebye, Eva B

    2014-11-01

    Ionization is a key factor in hERG K(+) channel blocking, and acids and zwitterions are known to be less probable hERG blockers than bases and neutral compounds. However, a considerable number of acidic compounds block hERG, and the physico-chemical attributes which discriminate acidic blockers from acidic non-blockers have not been fully elucidated. We propose a rule for prediction of hERG blocking by acids and zwitterionic ampholytes based on thresholds for only three descriptors related to acidity, size and reactivity. The training set of 153 acids and zwitterionic ampholytes was predicted with a concordance of 91% by a decision tree based on the rule. Two external validations were performed with sets of 35 and 48 observations, respectively, both showing concordances of 91%. In addition, a global QSAR model of hERG blocking was constructed based on a large diverse training set of 1374 chemicals covering all ionization classes, externally validated showing high predictivity and compared to the decision tree. The decision tree was found to be superior for the acids and zwitterionic ampholytes classes. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. Multidentate zwitterionic chitosan oligosaccharide modified gold nanoparticles: stability, biocompatibility and cell interactions

    NASA Astrophysics Data System (ADS)

    Liu, Xiangsheng; Huang, Haoyuan; Liu, Gongyan; Zhou, Wenbo; Chen, Yangjun; Jin, Qiao; Ji, Jian

    2013-04-01

    Surface engineering of nanoparticles plays an essential role in their colloidal stability, biocompatibility and interaction with biosystems. In this study, a novel multidentate zwitterionic biopolymer derivative is obtained from conjugating dithiolane lipoic acid and zwitterionic acryloyloxyethyl phosphorylcholine to the chitosan oligosaccharide backbone. Gold nanoparticles (AuNPs) modified by this polymer exhibit remarkable colloidal stabilities under extreme conditions including high salt conditions, wide pH range and serum or plasma containing media. The AuNPs also show strong resistance to competition from dithiothreitol (as high as 1.5 M). Moreover, the modified AuNPs demonstrate low cytotoxicity investigated by both MTT and LDH assays, and good hemocompatibility evaluated by hemolysis of human red blood cells. In addition, the intracellular fate of AuNPs was investigated by ICP-MS and TEM. It showed that the AuNPs are uptaken by cells in a concentration dependent manner, and they can escape from endosomes/lysosomes to cytosol and tend to accumulate around the nucleus after 24 h incubation but few of them are excreted out of the cells. Gold nanorods are also stabilized by this ligand, which demonstrates robust dispersion stability and excellent hemocompatibility. This kind of multidentate zwitterionic chitosan derivative could be widely used for stabilizing other inorganic nanoparticles, which will greatly improve their performance in a variety of bio-related applications.Surface engineering of nanoparticles plays an essential role in their colloidal stability, biocompatibility and interaction with biosystems. In this study, a novel multidentate zwitterionic biopolymer derivative is obtained from conjugating dithiolane lipoic acid and zwitterionic acryloyloxyethyl phosphorylcholine to the chitosan oligosaccharide backbone. Gold nanoparticles (AuNPs) modified by this polymer exhibit remarkable colloidal stabilities under extreme conditions including high salt

  5. Effect of dodecyl maltoside detergent on rhodopsin stability and function.

    PubMed

    Ramon, Eva; Marron, Jordi; del Valle, Luis; Bosch, Laia; Andrés, Anna; Manyosa, Joan; Garriga, Pere

    2003-12-01

    Detergent-solubilized bovine rhodopsin produces mixed detergent/lipid/protein micelles. The effect of dodecyl maltoside detergent on the thermal stability of dark-state rhodopsin, and upon formation of the different intermediates after rhodopsin photobleaching (metarhodopsin II and metarhodopsin III), and upon transducin activation has been studied. No significant effect is observed for the thermal stability of dark-state rhodopsin in the range of detergent concentrations studied, but a decrease in the stability of metarhodopsin II and an increase in metarhodopsin III formation is observed with decreasing detergent concentrations. The transducin activation process is also affected by the presence of detergent indicating that this process is dependent on the lipid micro-environment and membrane fluidity, and this stresses the importance of the native lipid environment in rhodopsin normal function.

  6. Improved antifouling performance of ultrafiltration membrane via preparing novel zwitterionic polyimide

    NASA Astrophysics Data System (ADS)

    Huang, Haitao; Yu, Jiayu; Guo, Hanxiang; Shen, Yibo; Yang, Fan; Wang, Han; Liu, Rong; Liu, Yang

    2018-01-01

    On the basis of the outstanding fouling resistance of zwitterionic polymers, an antifouling ultrafiltration membrane was fabricated through phase inversion induced by immersion precipitation method, directly using the novel zwitterionic polyimide (Z-PI), which was synthesized via a two-step procedure including polycondensation and quaternary amination reaction, as membrane material. The chemical structure and composition of the obtained polymer were confirmed by using FTIR, 1H NMR and XPS analysis, and its thermal stability was thoroughly characterized by TGA measurement, respectively. The introduction of zwitterionic groups into polyimide could effectively increase membrane pore size, porosity and wettability, and convert the membrane surface from hydrophobic to highly hydrophilic. As a result, Z-PI membrane displayed significantly improved water permeability compared with that of the reference polyimide (R-PI) membrane without having an obvious compromise in protein rejection. According to the static adsorption and dynamic cycle ultrafiltration experiments of bovine serum albumin (BSA) solution, Z-PI membrane exhibited better fouling resistant ability, especially irreversible fouling resistant ability, suggesting superior antifouling property and long-term performance stability. Moreover, Z-PI membrane had a water flux recovery ratio of 93.7% after three cycle of BSA solution filtration, whereas only about 68.5% was obtained for the control R-PI membrane. These findings demonstrated the advantages of Z-PI membrane material and aimed to provide a facile and scalable method for the large-scale preparation of low fouling ultrafiltration membranes for potential applications.

  7. Enzymatic detergent formulation containing amylase from Aspergillus niger: a comparative study with commercial detergent formulations.

    PubMed

    Mitidieri, Sydnei; Souza Martinelli, Anne Helene; Schrank, Augusto; Vainstein, Marilene Henning

    2006-07-01

    There is a wide range of biotechnological applications for amylases, including the textile, pharmaceutical, food and laundry industries. Hydrolytic enzymes are 100% biodegradable and enzymatic detergents can achieve effective cleaning with lukewarm water. Microorganisms and culture media were tested for amylase production and the best producer was Aspergillus niger L119 (3.9 U ml(-1) +/- 0.2) in submerged culture and its amylase demonstrated excellent activity at 50-55 degrees C and pH 4.0, remaining stable at 53 degrees C for up to 200 h. In order to establish the potential uses of this enzyme in detergents, different formulations were tested using the A. niger amylase extract. Enzyme activity was compared with three commercial formulations. The detergents are used in hospitals to clean surgical and endoscopy equipment. The presence of amylase in the formulation is because of its action within hospital drainage system, whether or not it has any function in cleaning the equipment.

  8. Quantum chemical characterization of zwitterionic structures: Supramolecular complexes for modifying the wettability of oil-water-limestone system.

    PubMed

    Lopez-Chavez, Ernesto; Garcia-Quiroz, Alberto; Gonzalez-Garcia, Gerardo; Orozco-Duran, Gabriela E; Zamudio-Rivera, Luis S; Martinez-Magadan, José M; Buenrostro-Gonzalez, Eduardo; Hernandez-Altamirano, Raul

    2014-06-01

    In this work, we present a quantum chemical study pertaining to some supramolecular complexes acting as wettability modifiers of oil-water-limestone system. The complexes studied are derived from zwitterionic liquids of the types N'-alkyl-bis, N-alquenil, N-cycloalkyl, N-amyl-bis-beta amino acid or salts acting as sparkling agents. We studied two molecules of zwitterionic liquids (ZL10 and ZL13), HOMO and LUMO levels, and the energy gap between them, were calculated, as well as the electron affinity (EA) and ionization potential (IP), chemical potential, chemical hardness, chemical electrophilicity index and selectivity descriptors such Fukui indices. In this work, electrochemical comparison was realized with cocamidopropyl betaine (CPB), which is a structure zwitterionic liquid type, nowadays widely applied in enhanced recovery processes. Copyright © 2014 Elsevier Inc. All rights reserved.

  9. Non-chapped, vertically well aligned titanium dioxide nanotubes fabricated by electrochemical etching

    NASA Astrophysics Data System (ADS)

    Loan Nguyen, Thu; Dieu Thuy Ung, Thi; Liem Nguyen, Quang

    2014-06-01

    This paper reports on the fabrication of non-chapped, vertically well aligned titanium dioxide nanotubes (TONTs) by using electrochemical etching method and further heat treatment. Very highly ordered metallic titanium nanotubes (TNTs) were formed by directly anodizing titanium foil at room temperature in an electrolyte composed of ammonium fluoride (NH4F), ethylene glycol (EG), and water. The morphology of as-formed TNTs is greatly dependent on the applied voltage, NH4F content and etching time. Particularly, we have found two interesting points related to the formation of TNTs: (i) the smooth surface without chaps of the largely etched area was dependent on the crystalline orientation of the titanium foil; and (ii) by increasing the anodizing potential from 15 V to 20 V, the internal diameter of TNT was increased from about 50 nm to 60 nm and the tube density decreased from 403 tubes μm-2 down to 339 tubes μm-2, respectively. For the anodizing duration from 1 h to 5 h, the internal diameter of each TNT was increased from ˜30 nm to 60 nm and the tube density decreased from 496 tubes μm-2 down to 403 tubes μm-2. After annealing at 400 °C in open air for 1 h, the TNTs were transformed into TONTs in anatase structure; further annealing at 600 °C showed the structural transformation from anatase to rutile as determined by Raman scattering spectroscopy.

  10. NMR spectroscopic and analytical ultracentrifuge analysis of membrane protein detergent complexes.

    PubMed

    Maslennikov, Innokentiy; Kefala, Georgia; Johnson, Casey; Riek, Roland; Choe, Senyon; Kwiatkowski, Witek

    2007-11-08

    Structural studies of integral membrane proteins (IMPs) are hampered by inherent difficulties in their heterologous expression and in the purification of solubilized protein-detergent complexes (PDCs). The choice and concentrations of detergents used in an IMP preparation play a critical role in protein homogeneity and are thus important for successful crystallization. Seeking an effective and standardized means applicable to genomic approaches for the characterization of PDCs, we chose 1D-NMR spectroscopic analysis to monitor the detergent content throughout their purification: protein extraction, detergent exchange, and sample concentration. We demonstrate that a single NMR measurement combined with a SDS-PAGE of a detergent extracted sample provides a useful gauge of the detergent's extraction potential for a given protein. Furthermore, careful monitoring of the detergent content during the process of IMP production allows for a high level of reproducibility. We also show that in many cases a simple sedimentation velocity measurement provides sufficient data to estimate both the oligomeric state and the detergent-to-protein ratio in PDCs, as well as to evaluate the homogeneity of the samples prior to crystallization screening. The techniques presented here facilitate the screening and selection of the extraction detergent, as well as help to maintain reproducibility in the detergent exchange and PDC concentration procedures. Such reproducibility is particularly important for the optimization of initial crystallization conditions, for which multiple purifications are routinely required.

  11. Synergistic effects on enantioselectivity of zwitterionic chiral stationary phases for separations of chiral acids, bases, and amino acids by HPLC.

    PubMed

    Hoffmann, Christian V; Pell, Reinhard; Lämmerhofer, Michael; Lindner, Wolfgang

    2008-11-15

    In an attempt to overcome the limited applicability scope of earlier proposed Cinchona alkaloid-based chiral weak anion exchangers (WAX) and recently reported aminosulfonic acid-based chiral strong cation exchangers (SCX), which are conceptionally restricted to oppositely charged solutes, their individual chiral selector (SO) subunits have been fused in a combinatorial synthesis approach into single, now zwitterionic, chiral SO motifs. The corresponding zwitterionic ion-exchange-type chiral stationary phases (CSPs) in fact combined the applicability spectra of the parent chiral ion exchangers allowing for enantioseparations of chiral acids and amine-type solutes in liquid chromatography using polar organic mode with largely rivaling separation factors as compared to the parent WAX and SCX CSPs. Furthermore, the application spectrum could be remarkably expanded to various zwitterionic analytes such as alpha- and beta-amino acids and peptides. A set of structurally related yet different CSPs consisting of either a quinine or quinidine alkaloid moiety as anion-exchange subunit and various chiral or achiral amino acids as cation-exchange subunits enabled us to derive structure-enantioselectivity relationships, which clearly provided strong unequivocal evidence for synergistic effects of the two oppositely charged ion-exchange subunits being involved in molecular recognition of zwitterionic analytes by zwitterionic SOs driven by double ionic coordination.

  12. [On health regulation of synthetic detergents].

    PubMed

    Frolova, A D; Sidorin, G I; Lukovnikova, L V; Skhodkina, N I; D'iakova, L I; Shaposhnikova, E S

    1999-01-01

    The authors present materials on hygienic regulation of such synthetic detergents as "Losk", "Dixan" in air of workplace and populated area, demonstrate results concerning evaluation of "Losk" effects. Discussion covers ways to evaluate chemical load caused by simultaneous emission of synthetic detergents from various environmental objects. For testing reliability of hygienic regulation for complex emission exemplified by "Losk", the authors suggest a model for express evaluation of complex exposure to chemicals.

  13. Impact of novel histone deacetylase inhibitors, CHAP31 and FR901228 (FK228), on adenovirus-mediated transgene expression.

    PubMed

    Taura, Kojiro; Yamamoto, Yuzo; Nakajima, Akio; Hata, Koichiro; Uchinami, Hiroshi; Yonezawa, Kei; Hatano, Etsuro; Nishino, Norikazu; Yamaoka, Yoshio

    2004-05-01

    Histone deacetylase inhibitors (HDIs) are known to enhance adenovirus (Ad)-mediated transgene expression. Recently, novel HDIs, including cyclic hydroxamic-acid-containing peptide 31 (CHAP31) and FR901228 (FK228), have been developed. The effects of these two novel HDIs on Ad-transduced or endogenous gene expression were investigated. Acetylation of core histones and the expression of the coxsackie and adenovirus receptor (CAR) in HDI-treated cells were examined using Western blot and a quantitative reverse transcription polymerase chain reaction (TaqMan RT-PCR), respectively. Their in vivo effect on adenoviral gene expression was investigated in BALB/c mice. Both compounds enhanced and prolonged Ad-mediated beta-galactosidase expression more effectively than did trichostatin A, a classic HDI. The same effect was observed in Ad-transduced heat shock protein 72 (HSP72), but not in hyperthermia-induced endogenous expression of HSP72, suggesting that the effect is specific for transduced gene expression. Hyperacetylation of core histones induced by HDIs was considered responsible for the augmentative effects of gene expression. Intravenous administration of either CHAP31 or FR901228 enhanced beta-galactosidase expression in mice infected with AdLacZ. CHAP31 and FR901228 amplified Ad-mediated transgene expression. The enhancement of transgene expression by HDIs may result in fewer vector doses for necessary gene expression, helping to alleviate disadvantages caused by Ad vectors. This could be a useful tool in overcoming current limitations of gene therapy using adenovirus vectors. Copyright 2004 John Wiley & Sons, Ltd.

  14. Synthetic detergents: 100 years of history.

    PubMed

    Kogawa, Ana Carolina; Cernic, Beatriz Gamberini; do Couto, Leandro Giovanni Domingos; Salgado, Hérida Regina Nunes

    2017-09-01

    In the year 2016 the synthetic detergents complete 100 years and in this story they evolved as cleaners. They are already part of the routine of thousands of people worldwide. For a higher power of cleaning of the detergent, today, are added phosphates, the main responsible for environmental problems. After 100 years of synthetic detergents, the effect of the combination of various cleaners on the environment is a gap. Legislation and guidelines about the other components of the formula of cleaners still missing. Even the term biodegradable can be best placed on the diversity of products currently entitled biodegradable. A lot attitudes can still be taken to continuously improve the relationship between the parties involved, animals, plants, waters and men, so that in another 100 years continues to exist this interaction with the environment without destroying it. The marketing used by synthetic detergent companies evolved a lot over the years and showed maturity to deal with changes in theories and strategies for promotion and even with the constant social reform that its consumer lived, accompanying them intelligently to be able to capture their changing needs and desires, and so assemble the best way to connect to them. This paper focuses on the subject synthetic detergents as well as (i) types and applications, (ii) threats, (iii) sustainability, (iv) legislation, (v) packaging and (vi) marketing strategies.

  15. Mitigation of Biofilm Development on Thin-Film Composite Membranes Functionalized with Zwitterionic Polymers and Silver Nanoparticles.

    PubMed

    Liu, Caihong; Faria, Andreia F; Ma, Jun; Elimelech, Menachem

    2017-01-03

    We demonstrate the functionalization of thin-film composite membranes with zwitterionic polymers and silver nanoparticles (AgNPs) for combating biofouling. Combining hydrophilic zwitterionic polymer brushes and biocidal AgNPs endows the membrane with dual functionality: antiadhesion and bacterial inactivation. An atom transfer radical polymerization (ATRP) reaction is used to graft zwitterionic poly(sulfobetaine methacrylate) (PSBMA) brushes to the membrane surface, while AgNPs are synthesized in situ through chemical reduction of silver. Two different membrane architectures (Ag-PSBMA and PSBMA-Ag TFC) are developed according to the sequence AgNPs, and PSBMA brushes are grafted on the membrane surface. A static adhesion assay shows that both modified membranes significantly reduced the adsorption of proteins, which served as a model organic foulant. However, improved antimicrobial activity is observed for PSBMA-Ag TFC (i.e., AgNPs on top of the polymer brush) in comparison to the Ag-PSBMA TFC membrane (i.e., polymer brush on top of AgNPs), indicating that architecture of the antifouling layer is an important factor in the design of zwitterion-silver membranes. Confocal laser scanning microscopy (CLSM) imaging indicated that PSBMA-Ag TFC membranes effectively inhibit biofilm formation under dynamic cross-flow membrane biofouling tests. Finally, we demonstrate the regeneration of AgNPs on the membrane after depletion of silver from the surface of the PSBMA-Ag TFC membrane.

  16. NMR spectroscopic and analytical ultracentrifuge analysis of membrane protein detergent complexes

    PubMed Central

    Maslennikov, Innokentiy; Kefala, Georgia; Johnson, Casey; Riek, Roland; Choe, Senyon; Kwiatkowski, Witek

    2007-01-01

    Background Structural studies of integral membrane proteins (IMPs) are hampered by inherent difficulties in their heterologous expression and in the purification of solubilized protein-detergent complexes (PDCs). The choice and concentrations of detergents used in an IMP preparation play a critical role in protein homogeneity and are thus important for successful crystallization. Results Seeking an effective and standardized means applicable to genomic approaches for the characterization of PDCs, we chose 1D-NMR spectroscopic analysis to monitor the detergent content throughout their purification: protein extraction, detergent exchange, and sample concentration. We demonstrate that a single NMR measurement combined with a SDS-PAGE of a detergent extracted sample provides a useful gauge of the detergent's extraction potential for a given protein. Furthermore, careful monitoring of the detergent content during the process of IMP production allows for a high level of reproducibility. We also show that in many cases a simple sedimentation velocity measurement provides sufficient data to estimate both the oligomeric state and the detergent-to-protein ratio in PDCs, as well as to evaluate the homogeneity of the samples prior to crystallization screening. Conclusion The techniques presented here facilitate the screening and selection of the extraction detergent, as well as help to maintain reproducibility in the detergent exchange and PDC concentration procedures. Such reproducibility is particularly important for the optimization of initial crystallization conditions, for which multiple purifications are routinely required. PMID:17988403

  17. Laundry detergent capsules and pediatric poisoning.

    PubMed

    Bonney, Asha G; Mazor, Suzan; Goldman, Ran D

    2013-12-01

    A 4-year-old girl was brought into the emergency department vomiting after having had ingested a laundry detergent capsule (LDC) from under the sink at her house. What is the risk of LDC poisoning? What can be done to treat these children? Laundry detergent capsules are relatively new to supermarket shelves in North America, and there has been an emergence of case reports in the literature describing LDC poisoning, which is worse than poisoning from other laundry detergents. Very little is known about the mechanisms causing these severe reactions, which include airway compromise and esophageal perforation, but the attractive appearance of these capsules and easy access at home has governments and health officials concerned about an increase in poisoning. No residual problems have been associated with these cases to date; however, further research is needed to assess long-term effects.

  18. Effects of wastewater sludge and its detergents on the stability of rotavirus

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

    Ward, R.L.; Ashley, C.S.

    1980-06-01

    Wastewater sludge reduced the heat required to inactivate rotavirus SA-11, and ionic detergents were identified as the sludge components responsible for this effect. A similar result was found previously with reovirus. The quantitative effects of individual ionic detergents on rotavirus and reovirus were very different, and rotavirus was found to be extremely sensitive to several of these detergents. However, neither virus was destabilized by nonionic detergents. On the contrary, rotavirus was stabilized by a nonionic detergent against the potent destabilizing effects of the ionic detergent sodium dodecyl sulfate. The destabilizing effects of both cationic and anionic detergents on rotavirus weremore » greatly altered by changes in the pH of the medium.« less

  19. Effect of ionic detergents, nonionic detergents, and chaotropic agents on polyphenol oxidase activity from dormant saffron (Crocus sativus L.) corms.

    PubMed

    Saeidian, Shahriar; Keyhani, Ezzatollah; Keyhani, Jacqueline

    2007-05-02

    Polyphenol oxidase (PPO; EC 1.14.18.1) catalyzes the hydroxylation of monophenols to o-diphenols (cresolase activity) and the oxidation of o-diphenols to o-quinones (catecholase activity), leading to browning in plants and produce. Further interest in the enzyme has been triggered by the active role that it plays in plant defense systems. PPO can be found in latent forms and is activated in vitro by various agents including urea, detergents, and proteases. The activation of PPO from several sources by sodium dodecyl sulfate (SDS) has been extensively investigated, but reports on the effect of other detergents or on the differential effect of detergents on each of PPO's activities are scarce. In addition, investigations on the enzyme in other plant parts besides fruits and vegetables are also scarce. Here, the effect of various detergents and chaotropic agents on PPO from dormant saffron (Crocus sativus L.) corm extract was investigated. SDS and sarkosyl activated the cresolase activity, while only SDS activated the catecholase activity. All other detergents tested, in milli- or micromolar concentrations, inhibited the cresolase activity but barely affected the catecholase activity. In contrast, urea and guanidine-HCl drastically inhibited the catecholase activity but moderately inhibited the cresolase activity. The same effects were obtained on the partially purified enzyme. Results identified a PPO, present in dormant corms, which was activated only by anionic detergents and was inhibited by other reputed activating agents such as urea. Results also emphasized the differences in structure and accessibility of the active sites for cresolase and catecholase activities.

  20. Effect of Detergent on Electrical Properties of Squid Axon Membrane

    PubMed Central

    Kishimoto, Uichiro; Adelman, William J.

    1964-01-01

    The effects of detergents on squid giant axon action and resting potentials as well as membrane conductances in the voltage clamp have been studied. Anionic detergents (sodium lauryl sulfate, 0.1 to 1.0 mM; dimethyl benzene sulfonate, 1 to 20 mM, pH 7.6) cause a temporary increase and a later decrease of action potential height and the value of the resting potential. Cationic detergent (cetyl trimethyl ammonium chloride, 6 x 10-5 M or more, pH 7.6) generally brings about immediate and irreversible decreases in the action and resting potentials. Non-ionic detergent (tween 80, 0.1 M, pH 7.6) causes a slight reversible reduction of action potential height without affecting the value of the resting potential. Both anionic and cationic detergents generally decrease the sodium and potassium conductances irreversibly. The effect of non-ionic detergent is to decrease the sodium conductance reversibly, leaving the potassium conductance almost unchanged. PMID:14158665

  1. Molecular size of the gamma-aminobutyric acidA receptor purified from mammalian cerebral cortex.

    PubMed

    Mamalaki, C; Barnard, E A; Stephenson, F A

    1989-01-01

    The hydrodynamic behaviour of both the soluble and purified gamma-aminobutyric acidA (GABAA) receptor of bovine or rat cerebral cortex has been investigated in solution in Triton X-100 or in 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulphonate (CHAPS). In all the hydrodynamic separations made, it was found that the binding activities for GABA, benzodiazepine, and (where detectable) t-butylbicyclophosphorothionate comigrated. Conditions were established for gel exclusion chromatography and for sucrose density gradient velocity sedimentation that maintain the GABAA receptor in a nonaggregated form. Using these conditions, the molecular weight of the bovine GABAA receptor in the above-mentioned detergents was calculated using the H2O/2H2O method. A value of Mr 230,000-240,000 was calculated for the bovine pure GABAA receptor purified in sodium deoxycholate/Triton X-100 media. A value of Mr 284,000-290,000 was calculated for the nonaggregated bovine or rat cortex receptor in CHAPS, but the Stokes radius is smaller in the latter than in the former medium and the detergent binding in CHAPS is underestimated. Thus the deduced Mr, 240,000, is the best estimate by this method.

  2. Hemocompatibility improvement of poly(ethylene terephthalate) via self-polymerization of dopamine and covalent graft of zwitterions.

    PubMed

    Cai, Xianmei; Yuan, Jiang; Chen, Shuangchun; Li, Pengfei; Li, Li; Shen, Jian

    2014-03-01

    Poly (ethylene terephthalate) (PET) has been widely adopted as a scaffold biomaterial, but further hemocompatibility improvement is still needed for wide biomedical applications. Inspired by the composition of adhesive proteins in mussels, we propose to use self-polymerized dopamine to form a surface-adherent polydopamine layer onto PET sheet, followed by Michael addition with N,N-dimethylethylenediamine (DMDA) to build tertiary amine, and final zwitterions(sulfobetaine and carboxybetaine) construction through ring-opening reaction. Physicochemical properties of substrates were demonstrated by water contact angle measurement, attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) and X-ray photoelectron spectroscopy (XPS). The hemocompatibility was evaluated by platelet adhesion, hemolytic, and protein adsorption. The results showed that the zwitterions immobilized PET endowed with improved resistance to nonspecific protein adsorption and platelet adhesion as well as nonhemolytic. The zwitterions with desirable hemocompatibility can be readily tailored to catheter for various biomedical applications. Copyright © 2013 Elsevier B.V. All rights reserved.

  3. Polysulfone and polyacrylate-based zwitterionic coatings for the prevention and easy removal of marine biofouling

    DOE PAGES

    Hibbs, Michael R.; Hernandez-Sanchez, Bernadette A.; Daniels, Justin; ...

    2015-09-07

    A series of polysulfone and polyacrylate-based zwitterionic coatings were prepared on epoxy-primed aluminum substrata and characterized for their antifouling (AF) and fouling-release (FR) properties towards marine bacteria, microalgae and barnacles. The zwitterionic polymer coatings provided minimal resistance against bacterial biofilm retention and microalgal cell attachment, but facilitated good removal of attached microbial biomass by exposure to water-jet apparatus generated hydrodynamic shearing forces. Increasing the ion content of the coatings improved the AF properties, but required a stronger adhesive bond to the epoxy-primed aluminum substratum to prevent coating swelling and dissolution. Grafted poly(sulfobetaine) (gpSBMA), the most promising zwitterionic coating identified frommore » microfouling evaluations, enabled the removal of four out of five barnacles reattached to its surface without incurring damage to their baseplates. As a result, this significant result indicated that gpSBMA relied predominately on its surface chemistry for its FR properties since it was very thin (~1–2 µm) relative to commercial coating standards (>200 µm).« less

  4. Effects of Lubrol detergents on adenylate cyclases.

    PubMed

    Bär, H P; Kulshrestha, S

    1975-04-01

    The nonionic detergent Lubrol WX showed diverse, concentration-dependent effects onbasal and stimulated adenylate cyclases. Above concentrations of 0.001-0.01% Lubrol WX, the basal activity of cyclase from Ehrlich ascites cells was inhibed about 50%, and that from rat fat cells was doubled. In both cases, hormonal sensitivity was lost at 0.01%. These effects were reversed upon dilution of the detergent. It is suggested that solubilization of adenylate cyclases at such low concentrations of Lubrol should be attempted since it is conceivable that loss of hormone sensitivity may then be reversible. Different Lubrol-type detergents may also offer centain advantages, since Lubrol PX effects were not identical with those of Lubrol WX.

  5. Anaerobic Biodegradation of Detergent Surfactants

    PubMed Central

    Merrettig-Bruns, Ute; Jelen, Erich

    2009-01-01

    Detergent surfactants can be found in wastewater in relevant concentrations. Most of them are known as ready degradable under aerobic conditions, as required by European legislation. Far fewer surfactants have been tested so far for biodegradability under anaerobic conditions. The natural environment is predominantly aerobic, but there are some environmental compartments such as river sediments, sub-surface soil layer and anaerobic sludge digesters of wastewater treatment plants which have strictly anaerobic conditions. This review gives an overview on anaerobic biodegradation processes, the methods for testing anaerobic biodegradability, and the anaerobic biodegradability of different detergent surfactant types (anionic, nonionic, cationic, amphoteric surfactants).

  6. Irritancy potential of 17 detergents used commonly by the Indian household.

    PubMed

    Austoria, A J; Lakshmi, Chembolli; Srinivas, C R; Anand, C V; Mathew, A C

    2010-01-01

    Detergents are used by almost every household in the developed and developing world. Soap and most detergents are anionic surfactants and attack the horny layer of the skin and increase its permeability with little or no inflammatory change and may result in hand eczema, which is very distressing and incapacitating. To evaluate the irritant potential of common household detergents (laundry and dish wash) used by the Indian population using a 24-hour patch test and to convincingly educate the patients on the detergents less likely to cause irritation in the particular individual. Seventeen commonly used detergents found in Indian market were included in the study, of which, 12 were laundry detergents (powders--seven, bar soap--five) and five were dish wash detergents (powder--one, liquid--one, bar soap--three). The irritant potential of the 17 detergents were evaluated in 30 volunteers. Thirty microliters of each of the detergent bar solutions, distilled water (negative control), and 20% SDS (positive control) were applied to Finn chambers with a micropipette and occluded for 24 hours. Erythema, scaling, and edema were graded in comparison to the reaction at the negative control site (distilled water) for each volunteer separately. The scoring of erythema/dryness and wrinkling on a 0-4 point scale and edema on another 0-4 point scale was based on the Draize scale. The pH of each of the detergent solutions was determined using litmus papers (Indikrom papers from Qualigens fine chemicals). The difference between detergents (F value) was significant for erythema/dryness and wrinkling (F = 3.374; p = 0.000), but not significant for edema (F = 1.297; p = 0.194). [Table 2] lists the means for erythema/dryness and wrinkling, and edema. The F value of the totals of the means for erythema/dryness and wrinkling and edema was significant (F = 2.495; p = 0.001). The pH of all the detergents was found to be alkaline except Pril utensil cleaner which tested acidic (pH 6). The

  7. Laundry detergent capsules and pediatric poisoning

    PubMed Central

    Bonney, Asha G.; Mazor, Suzan; Goldman, Ran D.

    2013-01-01

    Abstract Question A 4-year-old girl was brought into the emergency department vomiting after having had ingested a laundry detergent capsule (LDC) from under the sink at her house. What is the risk of LDC poisoning? What can be done to treat these children? Answer Laundry detergent capsules are relatively new to supermarket shelves in North America, and there has been an emergence of case reports in the literature describing LDC poisoning, which is worse than poisoning from other laundry detergents. Very little is known about the mechanisms causing these severe reactions, which include airway compromise and esophageal perforation, but the attractive appearance of these capsules and easy access at home has governments and health officials concerned about an increase in poisoning. No residual problems have been associated with these cases to date; however, further research is needed to assess long-term effects. PMID:24336541

  8. Detergents compared with each other and with antiseptics as skin 'degerming' agents.

    PubMed Central

    Lilly, H. A.; Lowbury, E. J.; Wilkins, M. D.

    1979-01-01

    Three detergent preparations (bar soap, 'Hibiscrub' base and 'LIC 76'), TWO ANTISEPTic preparations (0.5% chlorhexidine in 95% ethyl alcohol and an alcohol jelly, 'Alcogel'), and one antiseptic-detergent solution (4% chlorhexidine gluconate in a detergent base, 'Hibiscrub') were compared for their effectiveness, on a single use, in reducing the yield of bacteria from the hands of volunteers. The antiseptic and antiseptic--detergent preparations were more effective than the detergents, with a mean reduction in yield of skin bacteria of 96.0% after use of alcoholic chlorhexidine and of 81.2% after use of Hibiscrub. One of the detergents, LIC 76, appeared more effective than the others, causing a mean reduction in the yield of skin bacteria of 41.5%, compared with reductions of 4.6% by the Hibiscrub detergent base and an increase of 3.2% with bar soap; unlike the other detergents, LIC 76 was found to have appreciable bacteristatic and bactericidal properties. PMID:762408

  9. Correlating Detergent Fiber Analysis and Dietary Fiber Analysis Data for Corn Stover

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

    Wolfrum, E. J.; Lorenz, A. J.; deLeon, N.

    There exist large amounts of detergent fiber analysis data [neutral detergent fiber (NDF), acid detergent fiber (ADF), acid detergent lignin (ADL)] for many different potential cellulosic ethanol feedstocks, since these techniques are widely used for the analysis of forages. Researchers working in the area of cellulosic ethanol are interested in the structural carbohydrates in a feedstock (principally glucan and xylan), which are typically determined by acid hydrolysis of the structural fraction after multiple extractions of the biomass. These so-called dietary fiber analysis methods are significantly more involved than detergent fiber analysis methods. The purpose of this study was to determinemore » whether it is feasible to correlate detergent fiber analysis values to glucan and xylan content determined by dietary fiber analysis methods for corn stover. In the detergent fiber analysis literature cellulose is often estimated as the difference between ADF and ADL, while hemicellulose is often estimated as the difference between NDF and ADF. Examination of a corn stover dataset containing both detergent fiber analysis data and dietary fiber analysis data predicted using near infrared spectroscopy shows that correlations between structural glucan measured using dietary fiber techniques and cellulose estimated using detergent techniques, and between structural xylan measured using dietary fiber techniques and hemicellulose estimated using detergent techniques are high, but are driven largely by the underlying correlation between total extractives measured by fiber analysis and NDF/ADF. That is, detergent analysis data is correlated to dietary fiber analysis data for structural carbohydrates, but only indirectly; the main correlation is between detergent analysis data and solvent extraction data produced during the dietary fiber analysis procedure.« less

  10. Control of occupational asthma and allergy in the detergent industry.

    PubMed

    Sarlo, Katherine

    2003-05-01

    To provide an overview of how a comprehensive preclinical, clinical, and industrial hygiene program has been successfully used to control allergy and asthma to enzymes used in the detergent industry. The author performed a PubMed and ToxLine search of English-language articles with the keywords enzymes, occupational allergy, occupational asthma, detergent, and detergent industry from January 1, 1995, to January 1, 2002. Scientific meeting abstracts, books, and industry association papers on allergy and asthma in the detergent industry were also reviewed. In addition, the practical experience of one major detergent company was included in the review. All published work on this topic was reviewed, and the work that discussed the key highlights of control of occupational allergy and asthma to enzymes used in the detergent industry was selected for this review. The detergent industry has developed guidelines for the safety assessment of enzymes, control of exposure to enzymes, and medical surveillance of enzyme-exposed workers. Because of these guidelines, occupational allergy and asthma to enzymes used in the detergent industry have become uncommon events. Cases of disease have been documented in some manufacturing sites that have had poor adherence to the guidelines. Those manufacturing sites that have adhered to the guidelines have had few cases of allergy and asthma to enzymes among exposed workers. A review of medical data from these sites has shown that workers who have developed IgE antibody to enzymes can continue to work with enzymes and remain symptom free. Occupational allergy and asthma to enzymes used in the detergent industry have been successfully controlled via the use of preclinical, clinical, and industrial hygiene safety programs designed to minimize sensitization to enzymes and development of disease. The basic principles of these programs can be applied to other industries where occupational allergy and asthma to proteins are common.

  11. Evaluation of a New Lipase from Staphylococcus sp. for Detergent Additive Capability

    PubMed Central

    Chauhan, Mamta; Chauhan, Rajinder Singh; Garlapati, Vijay Kumar

    2013-01-01

    Lipases are the enzymes of choice for laundry detergent industries owing to their triglyceride removing ability from the soiled fabric which eventually reduces the usage of phosphate-based chemical cleansers in the detergent formulation. In the present study, a partially purified bacterial lipase from Staphylococcus arlettae JPBW-1 isolated from the rock salt mine has been assessed for its triglyceride removing ability by developing a presoak solution so as to use lipase as an additive in laundry detergent formulations. The effects of selected surfactants, commercial detergents, and oxidizing agents on lipase stability were studied in a preliminary evaluation for its further usage in the industrial environment. Partially purified lipase has shown good stability in presence of surfactants, commercial detergents, and oxidizing agents. Washing efficiency has been found to be enhanced while using lipase with 0.5% nonionic detergent than the anioinic detergent. The wash performance using 0.5% wheel with 40 U lipase at 40°C in 45 min results in maximum oil removal (62%) from the soiled cotton fabric. Hence, the present study opens the new era in enzyme-based detergent sector for formulation of chemical-free detergent using alkaline bacterial lipase. PMID:24106703

  12. Evaluation of a new lipase from Staphylococcus sp. for detergent additive capability.

    PubMed

    Chauhan, Mamta; Chauhan, Rajinder Singh; Garlapati, Vijay Kumar

    2013-01-01

    Lipases are the enzymes of choice for laundry detergent industries owing to their triglyceride removing ability from the soiled fabric which eventually reduces the usage of phosphate-based chemical cleansers in the detergent formulation. In the present study, a partially purified bacterial lipase from Staphylococcus arlettae JPBW-1 isolated from the rock salt mine has been assessed for its triglyceride removing ability by developing a presoak solution so as to use lipase as an additive in laundry detergent formulations. The effects of selected surfactants, commercial detergents, and oxidizing agents on lipase stability were studied in a preliminary evaluation for its further usage in the industrial environment. Partially purified lipase has shown good stability in presence of surfactants, commercial detergents, and oxidizing agents. Washing efficiency has been found to be enhanced while using lipase with 0.5% nonionic detergent than the anioinic detergent. The wash performance using 0.5% wheel with 40 U lipase at 40°C in 45 min results in maximum oil removal (62%) from the soiled cotton fabric. Hence, the present study opens the new era in enzyme-based detergent sector for formulation of chemical-free detergent using alkaline bacterial lipase.

  13. Highly Selective Enrichment of Glycopeptides Based on Zwitterionically Functionalized Soluble Nanopolymers

    NASA Astrophysics Data System (ADS)

    Cao, Weiqian; Huang, Jiangming; Jiang, Biyun; Gao, Xing; Yang, Pengyuan

    2016-07-01

    Efficient glycopeptides enrichment prior to mass spectrometry analysis is essential for glycoproteome study. ZIC-HILIC (zwitterionic hydrophilic interaction liquid chromatography) based glycopeptides enrichment approaches have been attracting more attention for several benefits like easy operating, high enrichment specificity and intact glycopeptide retained. In this study, Poly (amidoamine) dendrimer (PAMAM) was adopted for the synthesis of zwitterionically functionalized (ZICF) materials for glycopeptide enrichment. The multiple branched structure and good solubility of ZICF-PAMAM enables a sufficient interaction with glycopeptides. The ZICF-PAMAM combined with the FASP-mode enrichment strategy exhibits more superior performance compared with the existing methods. It has the minimum detectable concentration of femtomolar level and high recovery rate of over 90.01%, and can efficiently enrich glycopeptides from complex biological samples even for merely 0.1 μL human serum. The remarkable glycopeptides enrichment capacity of ZICF-PAMAM highlights the potential application in in-depth glycoproteome research, which may open up new opportunities for the development of glycoproteomics.

  14. Highly Selective Enrichment of Glycopeptides Based on Zwitterionically Functionalized Soluble Nanopolymers.

    PubMed

    Cao, Weiqian; Huang, Jiangming; Jiang, Biyun; Gao, Xing; Yang, Pengyuan

    2016-07-14

    Efficient glycopeptides enrichment prior to mass spectrometry analysis is essential for glycoproteome study. ZIC-HILIC (zwitterionic hydrophilic interaction liquid chromatography) based glycopeptides enrichment approaches have been attracting more attention for several benefits like easy operating, high enrichment specificity and intact glycopeptide retained. In this study, Poly (amidoamine) dendrimer (PAMAM) was adopted for the synthesis of zwitterionically functionalized (ZICF) materials for glycopeptide enrichment. The multiple branched structure and good solubility of ZICF-PAMAM enables a sufficient interaction with glycopeptides. The ZICF-PAMAM combined with the FASP-mode enrichment strategy exhibits more superior performance compared with the existing methods. It has the minimum detectable concentration of femtomolar level and high recovery rate of over 90.01%, and can efficiently enrich glycopeptides from complex biological samples even for merely 0.1 μL human serum. The remarkable glycopeptides enrichment capacity of ZICF-PAMAM highlights the potential application in in-depth glycoproteome research, which may open up new opportunities for the development of glycoproteomics.

  15. Characterization of the Protease Activity of Detergents: Laboratory Practicals for Studying the Protease Profile and Activity of Various Commercial Detergents

    ERIC Educational Resources Information Center

    Valls, Cristina; Pujadas, Gerard; Garcia-Vallve, Santi; Mulero, Miquel

    2011-01-01

    Detergent enzymes account for about 30% of the total worldwide production of enzymes and are one of the largest and most successful applications of modern industrial biotechnology. Proteases can improve the wash performance of household, industrial, and institutional laundry detergents used to remove protein-based stains such as blood, grass, body…

  16. Hollow Fiber Membrane Modification with Functional Zwitterionic Macromolecules for Improved Thromboresistance in Artificial Lungs

    PubMed Central

    Ye, Sang-Ho; Arazawa, David T.; Zhu, Yang; Shankarraman, Venkat; Malkin, Alexander D.; Kimmel, Jeremy D.; Gamble, Lara J.; Ishihara, Kazuhiko; Federspiel, William J.; Wagner, William R.

    2015-01-01

    Respiratory assist devices seek optimized performance in terms of gas transfer efficiency and thromboresistance to minimize device size and reduce complications associated with inadequate blood biocompatibility. The exchange of gas with blood occurs at the surface of the hollow fiber membranes (HFMs) used in these devices. In this study, three zwitterionic macromolecules were attached to HFM surfaces to putatively improve thromboresistance: (1) carboxyl-functionalized zwitterionic phosphorylcholine (PC) and (2) sulfobetaine (SB) macromolecules (mPC or mSB-COOH) prepared by a simple thiol-ene radical polymerization and (3) a low-molecular weight sulfobetaine (SB)-co-methacrylic acid (MA) block copolymer (SBMAb-COOH) prepared by reversible addition–fragmentation chain transfer (RAFT) polymerization. Each macromolecule type was covalently immobilized on an aminated commercial HFM (Celg-A) by a condensation reaction, and HFM surface composition changes were analyzed by X-ray photoelectron spectroscopy. Thrombotic deposition on the HFMs was investigated after contact with ovine blood in vitro. The removal of CO2 by the HFMs was also evaluated using a model respiratory assistance device. The HFMs conjugated with zwitterionic macromolecules (Celg-mPC, Celg-mSB, and Celg-SBMAb) showed expected increases in phosphorus or sulfur surface content. Celg-mPC and Celg-SBMAb experienced rates of platelet deposition significantly lower than those of unmodified (Celg-A, >95% reduction) and heparin-coated (>88% reduction) control HFMs. Smaller reductions were seen with Celg-mSB. The CO2 removal rate for Celg-SBMAb HFMs remained comparable to that of Celg-A. In contrast, the rate of removal of CO2 for heparin-coated HFMs was significantly reduced. The results demonstrate a promising approach to modifying HFMs using zwitterionic macromolecules for artificial lung devices with improved thromboresistance without degradation of gas transfer. PMID:25669307

  17. Diesel fuel detergent additive performance and assessment

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

    Vincent, M.W.; Papachristos, M.J.; Williams, D.

    Diesel fuel detergent additives are increasingly linked with high quality automotive diesel fuels. Both in Europe and in the USA, field problems associated with fuel injector coking or fouling have been experienced. In Europe indirect injection (IDI) light duty engines used in passenger cars were affected, while in the USA, a direct injection (DI) engine in heavy duty truck applications experienced field problems. In both cases, a fuel additive detergent performance test has evolved using an engine linked with the original field problem, although engine design modifications employed by the manufacturers have ensured improved operation in service. Increasing awareness ofmore » the potential for injector nozzle coking to cause deterioration in engine performance is coupled with a need to meet ever more stringent exhaust emissions legislation. These two requirements indicate that the use of detergency additives will continue to be associated with high quality diesel fuels. The paper examines detergency performance evaluated in a range of IDI and DI engines and correlates performance in the two most widely recognised test engines, namely the Peugeot 1.9 litre IDI, and Cummins L10 DI engines. 17 refs., 18 figs., 5 tabs.« less

  18. Surface charge control for zwitterionic polymer brushes: Tailoring surface properties to antifouling applications.

    PubMed

    Guo, Shanshan; Jańczewski, Dominik; Zhu, Xiaoying; Quintana, Robert; He, Tao; Neoh, Koon Gee

    2015-08-15

    Electrostatic interactions play an important role in adhesion phenomena particularly for biomacromolecules and microorganisms. Zero charge valence of zwitterions has been claimed as the key to their antifouling properties. However, due to the differences in the relative strength of their acid and base components, zwitterionic materials may not be charge neutral in aqueous environments. Thus, their charge on surfaces should be further adjusted for a specific pH environment, e.g. physiological pH typical in biomedical applications. Surface zeta potential for thin polymeric films composed of polysulfobetaine methacrylate (pSBMA) brushes is controlled through copolymerizing zwitterionic SBMA and cationic methacryloyloxyethyltrimethyl ammonium chloride (METAC) via surface-initiated atom transfer polymerization. Surface properties including zeta potential, roughness, free energy and thickness are measured and the antifouling performance of these surfaces is assessed. The zeta potential of pSBMA brushes is -40 mV across a broad pH range. By adding 2% METAC, the zeta potential of pSBMA can be tuned to zero at physiological pH while minimally affecting other physicochemical properties including dry brush thickness, surface free energy and surface roughness. Surfaces with zero and negative zeta potential best resist fouling by bovine serum albumin, Escherichia coli and Staphylococcus aureus. Surfaces with zero zeta potential also reduce fouling by lysozyme more effectively than surfaces with negative and positive zeta potential. Copyright © 2015 Elsevier Inc. All rights reserved.

  19. Detergent Disposal into Our Environmentand Its Impact on Marine Microbes

    NASA Astrophysics Data System (ADS)

    Effendi, I.; Nedi, S.; Ellizal; Nursyirwani; Feliatra; Fikar; Tanjung; Pakpahan, R.; Pratama

    2017-12-01

    Detergents figure in an extensive array of industrial and home cleaning applications, released into the flow of wastewater coming from the home, can far-reaching environmental impacts. Microorganisms are crucial to nutrient recycling in ecosystems as they act as decomposers, pathogen, antibiotic producer, biodegradation of pollutants etc. The research is aimed to examine effect detergent disposal to bacterial population growth in marine environment both in vitro and in situ condition. Seawater samples were collected from Sungai Kayu Ara Village, and Dumai River estuary, Siak Regency and Dumai City, Riau Province. Experimental method with complete randomized design (RAL) 2 (two) factors; a detergent brand (a1: ATTACK, a2; RINSO and a3; SURF) and b concentration of detergent concentration with 5 (five) concentration level, b1 (0%) as control, b2 (0.3%), b3 (0.6%), b4 (0.9%) and b5 (1.2%) wass applied. The study showed that there was an effect of detergent addition, periode of exposure, and doses to the growth of bacterial population both in vitro and in situ conditions. The higher levels of detergent in the water column and the longer contamination duration, causing more and more depressed bacterial populations. It is suggested to run a further research on identification, and growth optimatioan of the species capable of degrading detergent.

  20. The relative abundance of dimethylsulfoniopropionate (DMSP) among other zwitterions in branching coral at Heron Island, southern Great Barrier Reef.

    PubMed

    Swan, Hilton B; Deschaseaux, Elisabeth S M; Jones, Graham B; Eyre, Bradley D

    2017-07-01

    Dimethylsulfoniopropionate (DMSP) and eleven other target zwitterions were quantified in the branch tips of six Acropora species and Stylophora pistillata hard coral growing on the reef flat surrounding Heron Island in the southern Great Barrier Reef (GBR), Australia. Hydrophilic interaction liquid chromatography mass spectrometry (HILIC-MS) was used for sample analysis with isotope dilution MS applied to quantify DMSP. The concentration of DMSP was ten times greater in A. aspera than A. valida, with this difference being maintained throughout the spring, summer and winter seasons. In contrast, glycine betaine was present in significantly higher concentrations in these species during the summer than the winter. Exposure of branch tips of A. aspera to air and hypo-saline seawater for up to 1 h did not alter the concentrations of DMSP present in the coral when compared with control samples. DMSP was the most abundant target zwitterion in the six Acropora species examined, ranging from 44-78% of all target zwitterions in A. millepora and A. aspera, respectively. In contrast, DMSP only accounted for 7% in S. pistillata, with glycine betaine and stachydrine collectively accounting for 88% of all target zwitterions in this species. The abundance of DMSP in the six Acropora species examined points to Acropora coral being an important source for the biogeochemical cycling of sulfur throughout the GBR, since this reef-building branching coral dominates the coral cover of the GBR. Graphical Abstract HILIC-MS extracted ion chromatogram showing zwitterionic metabolites from the branching coral Acropora isopora.

  1. Surveillance of paediatric exposures to liquid laundry detergent pods in Italy

    PubMed Central

    Settimi, Laura; Giordano, Felice; Lauria, Laura; Celentano, Anna; Sesana, Fabrizio; Davanzo, Franca

    2018-01-01

    Objective To analyse paediatric exposures to pod and traditional laundry detergents in Italy and changes in exposure trends. Methods Analyses of a series of patients aged <5 years and exposed to laundry detergents between September 2010 and June 2015, identified by the National Poison Control in Milan. Results In comparison with patients exposed to traditional laundry detergents (n=1150), a higher proportion of those exposed to pods (n=1649) were managed in hospital (68% vs 42%), had clinical effects (75% vs 22%) and moderate/high severity outcomes (13% vs <1%). Exposure rates were stable over time for traditional detergents (average 0.65 cases/day), but an abrupt decline in major company pods was seen in December 2012, 4 months after the introduction of opaque outer packaging (from 1.03 to 0.36 cases/day and from 1.88 to 0.86 cases/million units sold). The odds of clinical effects was higher for exposure to pods than for traditional detergents (OR=10.8; 95% CI 9.0 to 12.9). Among patients exposed to pods, the odds of moderate/high severity outcomes was four times higher for children aged <1 years than for the other age groups (OR=3.9; 95% CI 2.2 to 7.0). Ten children exposed to laundry detergent pods had high severity outcomes while no children exposed to traditional laundry detergents developed high severity effects. Conclusions The study confirms that exposure to laundry detergent pods is more dangerous than exposure to traditional detergents. In Italy, 4 months after the introduction of opaque outer packaging by a major company, product-specific exposure rates decreased sharply, suggesting that reducing visibility of laundry detergent pods may be an effective preventive measure. Further efforts are needed to improve safety. PMID:28188147

  2. Biochemical characterization of detergent-resistant membranes: a systematic approach

    PubMed Central

    Babiychuk, Eduard B.; Draeger, Annette

    2006-01-01

    Lateral segregation of cholesterol- and sphingomyelin-rich rafts and glycerophospholipid-containing non-raft microdomains has been proposed to play a role in a variety of biological processes. The most compelling evidence for membrane segregation is based on the observation that extraction with non-ionic detergents leads to solubilization of a subset of membrane components only. However, one decade later, a large body of inconsistent detergent-extraction data is threatening the very concept of membrane segregation. We have assessed the validity of the existing paradigms and we show the following. (i) The localization of a membrane component within a particular fraction of a sucrose gradient cannot be taken as a yardstick for its solubility: a variable localization of the DRMs (detergent-resistant membranes) in sucrose gradients is the result of complex associations between the membrane skeleton and the lipid bilayer. (ii) DRMs of variable composition can be generated by using a single detergent, the increasing concentration of which gradually extracts one protein/lipid after another. Therefore any extraction pattern obtained by a single concentration experiment is bound to be ‘investigator-specific’. It follows that comparison of DRMs obtained by different detergents in a single concentration experiment is prone to misinterpretations. (iii) Depletion of cholesterol has a graded effect on membrane solubility. (iv) Differences in detergent solubility of the members of the annexin protein family arise from their association with chemically different membrane compartments; however, these cannot be attributed to the ‘brick-like’ raft-building blocks of fixed size and chemical composition. Our findings demonstrate a need for critical re-evaluation of the accumulated detergent-extraction data. PMID:16608442

  3. Biochemical characterization of detergent-resistant membranes: a systematic approach.

    PubMed

    Babiychuk, Eduard B; Draeger, Annette

    2006-08-01

    Lateral segregation of cholesterol- and sphingomyelin-rich rafts and glycerophospholipid-containing non-raft microdomains has been proposed to play a role in a variety of biological processes. The most compelling evidence for membrane segregation is based on the observation that extraction with non-ionic detergents leads to solubilization of a subset of membrane components only. However, one decade later, a large body of inconsistent detergent-extraction data is threatening the very concept of membrane segregation. We have assessed the validity of the existing paradigms and we show the following. (i) The localization of a membrane component within a particular fraction of a sucrose gradient cannot be taken as a yardstick for its solubility: a variable localization of the DRMs (detergent-resistant membranes) in sucrose gradients is the result of complex associations between the membrane skeleton and the lipid bilayer. (ii) DRMs of variable composition can be generated by using a single detergent, the increasing concentration of which gradually extracts one protein/lipid after another. Therefore any extraction pattern obtained by a single concentration experiment is bound to be 'investigator-specific'. It follows that comparison of DRMs obtained by different detergents in a single concentration experiment is prone to misinterpretations. (iii) Depletion of cholesterol has a graded effect on membrane solubility. (iv) Differences in detergent solubility of the members of the annexin protein family arise from their association with chemically different membrane compartments; however, these cannot be attributed to the 'brick-like' raft-building blocks of fixed size and chemical composition. Our findings demonstrate a need for critical re-evaluation of the accumulated detergent-extraction data.

  4. Matrix composition and mechanics of decellularized lung scaffolds.

    PubMed

    Petersen, Thomas H; Calle, Elizabeth A; Colehour, Maegen B; Niklason, Laura E

    2012-01-01

    The utility of decellularized native tissues for tissue engineering has been widely demonstrated. Here, we examine the production of decellularized lung scaffolds from native rodent lung using two different techniques, principally defined by use of either the detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) or sodium dodecyl sulfate (SDS). All viable cellular material is removed, including at least 99% of DNA. Histochemical staining and mechanical testing indicate that collagen and elastin are retained in the decellularized matrices with CHAPS-based decellularization, while SDS-based decellularization leads to loss of collagen and decline in mechanical strength. Quantitative assays confirm that most collagen is retained with CHAPS treatment but that about 80% of collagen is lost with SDS treatment. In contrast, for both detergent methods, at least 60% of elastin content is lost along with about 95% of native proteoglycan content. Mechanical testing of the decellularized scaffolds indicates that they are mechanically similar to native lung using CHAPS decellularization, including retained tensile strength and elastic behavior, demonstrating the importance of collagen and elastin in lung mechanics. With SDS decellularization, the mechanical integrity of scaffolds is significantly diminished with some loss of elastic function as well. Finally, a simple theoretical model of peripheral lung matrix mechanics is consonant with our experimental findings. This work demonstrates the feasibility of producing a decellularized lung scaffold that can be used to study lung matrix biology and mechanics, independent of the effects of cellular components. Copyright © 2011 S. Karger AG, Basel.

  5. 21 CFR 740.17 - Foaming detergent bath products.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 7 2014-04-01 2014-04-01 false Foaming detergent bath products. 740.17 Section 740.17 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) COSMETICS COSMETIC PRODUCT WARNING STATEMENTS Warning Statements § 740.17 Foaming detergent bath products...

  6. 21 CFR 740.17 - Foaming detergent bath products.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 7 2013-04-01 2013-04-01 false Foaming detergent bath products. 740.17 Section 740.17 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) COSMETICS COSMETIC PRODUCT WARNING STATEMENTS Warning Statements § 740.17 Foaming detergent bath products...

  7. 21 CFR 740.17 - Foaming detergent bath products.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 7 2011-04-01 2010-04-01 true Foaming detergent bath products. 740.17 Section 740.17 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) COSMETICS COSMETIC PRODUCT WARNING STATEMENTS Warning Statements § 740.17 Foaming detergent bath products...

  8. 21 CFR 740.17 - Foaming detergent bath products.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 7 2010-04-01 2010-04-01 false Foaming detergent bath products. 740.17 Section 740.17 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) COSMETICS COSMETIC PRODUCT WARNING STATEMENTS Warning Statements § 740.17 Foaming detergent bath products...

  9. 21 CFR 740.17 - Foaming detergent bath products.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 7 2012-04-01 2012-04-01 false Foaming detergent bath products. 740.17 Section 740.17 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) COSMETICS COSMETIC PRODUCT WARNING STATEMENTS Warning Statements § 740.17 Foaming detergent bath products...

  10. Surface modification of silicon wafer by grafting zwitterionic polymers to improve its antifouling property

    NASA Astrophysics Data System (ADS)

    Sun, Yunlong; Chen, Changlin; Xu, Heng; Lei, Kun; Xu, Guanzhe; Zhao, Li; Lang, Meidong

    2017-10-01

    Silicon (111) wafer was modified by triethoxyvinylsilane containing double bond as an intermedium, and then P4VP (polymer 4-vinyl pyridine) brush was "grafted" onto the surface of silicon wafer containing reactive double bonds by adopting the "grafting from" way and Si-P4VP substrate (silicon wafer grafted by P4VP) was obtained. Finally, P4VP brush of Si-P4VP substrate was modified by 1,3-propanesulfonate fully to obtain P4VP-psl brush (zwitterionic polypyridinium salt) and the functional Si-P4VP-psl substrate (silicon wafer grafted by zwitterionic polypyridinium salt based on polymer 4-vinyl pyridine) was obtained successfully. The antifouling property of the silicon wafer, the Si-P4VP substrate and the Si-P4VP-psl substrate was investigated by using bovine serum albumin, mononuclear macrophages (RAW 264.7) and Escherichia coli (E. coli) ATTC25922 as model bacterium. The results showed that compared with the blank sample-silicon wafer, the Si-P4VP-psl substrate had excellent anti-adhesion ability against bovine serum albumin, cells and bacterium, due to zwitterionic P4VP-psl brush (polymer 4-vinyl pyridine salt) having special functionality like antifouling ability on biomaterial field.

  11. Erythrocyte hemolysis by detergents.

    PubMed

    Chernitsky, E A; Senkovich, O A

    1997-01-01

    The numbers of Triton X-100 and sodium dodecyl sulfate molecules required to form respective pores were estimated from the relationship between the detergent concentrations and the rates of fast and slow hemolysis components. It has been found that the slow hemolysis component evoked by Triton X-100 is related to the existence of two different pores. It is shown that the fast hemolysis component induced by sodium dodecyl sulfate is associated with the modification of phosphatidylcholine which determines the break in the Arrhenius plots of the hemolysis rate within the range of 20 degrees C. The shape of hemolysis kinetic curves and the dependence of hemolytic parameters on the detergent concentration and temperature are discussed based on the concept of hemolysis caused by the formation of pores in various membrane lipid regions and by releasing vesicles from erythrocytes.

  12. 40 CFR 80.161 - Detergent additive certification program.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... the subject detergent additive in each test fuel, and the corresponding test results (percent flow... penalties in §§ 80.169 and 80.172. (iii) If both parties submit the required information, EPA will evaluate... customers who use the disqualified detergent. Failure to do so may subject the certifier to liabilities for...

  13. Efficient and Tunable Three-Dimensional Functionalization of Fully Zwitterionic Antifouling Surface Coatings.

    PubMed

    Lange, Stefanie C; van Andel, Esther; Smulders, Maarten M J; Zuilhof, Han

    2016-10-11

    To enhance the sensitivity and selectivity of surface-based (bio)sensors, it is of crucial importance to diminish background signals that arise from the nonspecific binding of biomolecules, so-called biofouling. Zwitterionic polymer brushes have been shown to be excellent antifouling materials. However, for sensing purposes, antifouling does not suffice but needs to be combined with the possibility to efficiently modify the brush with recognition units. So far this has been achieved only at the expense of either antifouling properties or binding capacity. Herein we present a conceptually new approach by integrating both characteristics into a single tailor-made monomer: a novel sulfobetaine-based zwitterionic monomer equipped with a clickable azide moiety. Copolymerization of this monomer with a well-established standard sulfobetaine monomer results in highly antifouling surface coatings with a large yet tunable number of clickable groups present throughout the entire brush. Subsequent functionalization of the azido brushes via widely used strain-promoted alkyne azide click reactions yields fully zwitterionic 3D-functionalized coatings with a recognition unit of choice that can be tailored for any specific application. Here we show a proof of principle with biotin-functionalized brushes on Si 3 N 4 that combine excellent antifouling properties with specific avidin binding from a protein mixture. The signal-to-noise ratio is significantly improved over that of traditional chain-end modification of sulfobetaine polymer brushes, even if the azide content is lowered to 1%. This therefore offers a viable approach to the development of biosensors with greatly enhanced performance on any surface.

  14. Modelling zwitterions in solution: 3-fluoro-γ-aminobutyric acid (3F-GABA).

    PubMed

    Cao, Jie; Bjornsson, Ragnar; Bühl, Michael; Thiel, Walter; van Mourik, Tanja

    2012-01-02

    The conformations and relative stabilities of folded and extended 3-fluoro-γ-aminobutyric acid (3F-GABA) conformers were studied using explicit solvation models. Geometry optimisations in the gas phase with one or two explicit water molecules favour folded and neutral structures containing intramolecular NH···O-C hydrogen bonds. With three or five explicit water molecules zwitterionic minima are obtained, with folded structures being preferred over extended conformers. The stability of folded versus extended zwitterionic conformers increases on going from a PCM continuum solvation model to the microsolvated complexes, though extended structures become less disfavoured with the inclusion of more water molecules. Full explicit solvation was studied with a hybrid quantum-mechanical/molecular-mechanical (QM/MM) scheme and molecular dynamics simulations, including more than 6000 TIP3P water molecules. According to free energies obtained from thermodynamic integration at the PM3/MM level and corrected for B3LYP/MM total energies, the fully extended conformer is more stable than folded ones by about -4.5 kJ mol(-1). B3LYP-computed (3)J(F,H) NMR spin-spin coupling constants, averaged over PM3/MM-MD trajectories, agree best with experiment for this fully extended form, in accordance with the original NMR analysis. The seeming discrepancy between static PCM calculations and experiment noted previously is now resolved. That the inexpensive semiempirical PM3 method performs so well for this archetypical zwitterion is encouraging for further QM/MM studies of biomolecular systems. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. A New Approach to Non-Coordinating Anions: Lewis Acid Enhancement of Porphyrin Metal Centers in a Zwitterionic Metal$-$Organic Framework

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

    Johnson, Jacob A.; Petersen, Brenna M.; Kormos, Attila

    Here, we describe a new strategy to generate non-coordinating anions using zwitterionic metal–organic frameworks (MOFs). By assembly of anionic inorganic secondary building blocks (SBUs) ([In(CO 2) 4] $-$) with cationic metalloporphyrin-based organic linkers, we prepared zwitterionic MOFs in which the complete internal charge separation effectively prevents the potential binding of the counteranion to the cationic metal center. We demonstrate the enhanced Lewis acidity of Mn III- and Fe III-porphyrins in the zwitterionic MOFs in three representative electrocyclization reactions: [2 + 1] cycloisomerization of enynes, [3 + 2] cycloaddition of aziridines and alkenes, and [4 + 2] hetero-Diels–Alder cycloaddition of aldehydesmore » with dienes. Lastly, this work paves a new way to design functional MOFs for tunable chemical catalysis.« less

  16. The Effect of Detergents on the Morphology and Immunomodulatory Activity of Malassezia furfur

    PubMed Central

    Kim, Su-Han; Ko, Hyun-Chang; Kwon, Kyung-Sool; Oh, Chang-Keun

    2009-01-01

    Background Several workers have found that Malassezia are capable of suppressing cytokine release and downregulating the phagocytic function of monocytes. But lipid-depleted Malassezia furfur (M. furfur) extracts have also been shown to induce increased production of TNF-α, IL-6 and IL-1β in monocytes. We thought that the detergents in shampoos or soaps could change the composition of the lipid in the M. furfur cell wall. Objective We studied whether detergents affect the morphology of M. furfur and if the inflammatory cytokine profiles change in the monocytes treated with detergent-treated M. furfur. Methods Commonly used detergents such as sodium lauryl sulfate, ammonium lauryl sulfate and tween-80 were respectively added to the modified Leeming-Notman's media. M. furfur was cultivated in each media (detergent-added or untreated). Thereafter, the surface morphology of the yeast was evaluated by scanning and transmission electron microscopy. The cytokine profiles of monocytes, which were treated by M. furfur with or without detergents, were also evaluated. Results The detergent-treated M. furfur were similar to the lipid-extracted form of M. furfur on the electron microscopic study, with a recessed, withered surface and with thinner and rather electron transparent cell walls than the detergent-untreated M. furfur. The levels of TNF-α were higher in monocytes treated with detergent-treated Malassezia than that in the monocytes treated with the detergent-untreated Malassezia (p<0.05). Conclusion According to the findings in this study, it could be inferred that the detergents in shampoos or soaps affect the lipid layers of the Malassezia cell wall and these lipid-extracted Malassezia induce or aggravate some inflammatory conditions. But to correlate the relationship between detergents and Malassezia-associated diseases, in vivo experiments that will focus on short-term contact with detergents in real life conditions should be done. PMID:20523770

  17. The Effect of Detergents on the Morphology and Immunomodulatory Activity of Malassezia furfur.

    PubMed

    Kim, Su-Han; Ko, Hyun-Chang; Kim, Moon-Bum; Kwon, Kyung-Sool; Oh, Chang-Keun

    2009-05-01

    Several workers have found that Malassezia are capable of suppressing cytokine release and downregulating the phagocytic function of monocytes. But lipid-depleted Malassezia furfur (M. furfur) extracts have also been shown to induce increased production of TNF-alpha, IL-6 and IL-1beta in monocytes. We thought that the detergents in shampoos or soaps could change the composition of the lipid in the M. furfur cell wall. We studied whether detergents affect the morphology of M. furfur and if the inflammatory cytokine profiles change in the monocytes treated with detergent-treated M. furfur. Commonly used detergents such as sodium lauryl sulfate, ammonium lauryl sulfate and tween-80 were respectively added to the modified Leeming-Notman's media. M. furfur was cultivated in each media (detergent-added or untreated). Thereafter, the surface morphology of the yeast was evaluated by scanning and transmission electron microscopy. The cytokine profiles of monocytes, which were treated by M. furfur with or without detergents, were also evaluated. The detergent-treated M. furfur were similar to the lipid-extracted form of M. furfur on the electron microscopic study, with a recessed, withered surface and with thinner and rather electron transparent cell walls than the detergent-untreated M. furfur. The levels of TNF-alpha were higher in monocytes treated with detergent-treated Malassezia than that in the monocytes treated with the detergent-untreated Malassezia (p<0.05). According to the findings in this study, it could be inferred that the detergents in shampoos or soaps affect the lipid layers of the Malassezia cell wall and these lipid-extracted Malassezia induce or aggravate some inflammatory conditions. But to correlate the relationship between detergents and Malassezia-associated diseases, in vivo experiments that will focus on short-term contact with detergents in real life conditions should be done.

  18. Synthesis and Properties of Dodecyl Trehaloside Detergents for Membrane Protein Studies

    PubMed Central

    Tao, Houchao; Fu, Yu; Thompson, Aaron; Lee, Sung Chang; Mahoney, Nicholas; Stevens, Raymond C.; Zhang, Qinghai

    2012-01-01

    Sugar-based detergents, mostly derived from maltose or glucose, prevail in the extraction, solubilization, stabilization and crystallization of membrane proteins. Inspired by the broad use of trehalose for protecting biological macromolecules and lipid bilayer structures, we synthesized new trehaloside detergents for potential applications in membrane protein research. We devised an efficient synthesis of four dodecyl trehalosides, each with the 12-carboned alkyl chain attached to different hydroxyl groups of trehalose, thus presenting a structurally diverse but related family of detergents. The detergent physical properties, including solubility, hydrophobicity, critical micelle concentration (CMC) and size of micelles, were evaluated and compared with the most popular maltoside analog, β- D-dodecylmaltoside (DDM), which varied from each other due to distinct molecular geometries and possible polar group interactions in resulting micelles. Crystals of 2-dodecyl trehaloside (2-DDTre) were also obtained in methanol, and the crystal packing revealed multiple H-bonded interactions among adjacent trehalose groups. The few trehaloside detergents were tested for the solubilization and stabilization of the nociceptin/orphanin FQ peptide receptor (ORL1) and MsbA, which belong to the G-protein coupled receptor (GPCR) and ATP-binding cassette transporter families, respectively. Our results demonstrated the utility of trehaloside detergents as membrane protein solubilization reagents with the optimal detergents being protein dependent. Continuing development and investigations of trehaloside detergents are attractive given their interesting and unique chemical-physical properties and potential interactions with membrane lipids. PMID:22780816

  19. Soaps and detergents: understanding their composition and effect.

    PubMed

    Kirsner, R S; Froelich, C W

    1998-03-01

    Soaps have been used for thousands of years as part of religious ceremonies and daily life. Derived from fatty acids or triglycerides (fats or oils) into their alkali derivatives through a process called saponification, soaps are important for healthcare professionals in preventing the spread of disease. Partly due to their alkaline nature, soaps are limited by their irritancy to the skin and their tendency to form insoluble and inactive salts when combined with either hard water or sea water. Therefore, soap alternatives or synthetic detergents have been developed. Detergents are classified into four groups: anionic, cationic, amphoteric, and non-ionic. These four groups are based on the hydrophilic qualities and surfactants they possess. Each group has characteristics that pertain to its main uses, irritancy, and toxicity. Understanding soaps and detergents may assist clinicians in making intelligent choices when using these agents on their patients as either skin cleansers or wound cleansers. Understanding the characteristics of soaps and detergents is especially important when dealing with at-risk patients such as the elderly.

  20. Bypassing Protein Corona Issue on Active Targeting: Zwitterionic Coatings Dictate Specific Interactions of Targeting Moieties and Cell Receptors.

    PubMed

    Safavi-Sohi, Reihaneh; Maghari, Shokoofeh; Raoufi, Mohammad; Jalali, Seyed Amir; Hajipour, Mohammad J; Ghassempour, Alireza; Mahmoudi, Morteza

    2016-09-07

    Surface functionalization strategies for targeting nanoparticles (NP) to specific organs, cells, or organelles, is the foundation for new applications of nanomedicine to drug delivery and biomedical imaging. Interaction of NPs with biological media leads to the formation of a biomolecular layer at the surface of NPs so-called as "protein corona". This corona layer can shield active molecules at the surface of NPs and cause mistargeting or unintended scavenging by the liver, kidney, or spleen. To overcome this corona issue, we have designed biotin-cysteine conjugated silica NPs (biotin was employed as a targeting molecule and cysteine was used as a zwitterionic ligand) to inhibit corona-induced mistargeting and thus significantly enhance the active targeting capability of NPs in complex biological media. To probe the targeting yield of our engineered NPs, we employed both modified silicon wafer substrates with streptavidin (i.e., biotin receptor) to simulate a target and a cell-based model platform using tumor cell lines that overexpress biotin receptors. In both cases, after incubation with human plasma (thus forming a protein corona), cellular uptake/substrate attachment of the targeted NPs with zwitterionic coatings were significantly higher than the same NPs without zwitterionic coating. Our results demonstrated that NPs with a zwitterionic surface can considerably facilitate targeting yield of NPs and provide a promising new type of nanocarriers in biological applications.

  1. Bioinspired Zwitterionic Surface Coatings with Robust Photostability and Fouling Resistance.

    PubMed

    Huang, Chun-Jen; Chu, Sz-Hau; Wang, Lin-Chuan; Li, Chien-Hung; Lee, T Randall

    2015-10-28

    Great care has been paid to the biointerface between a bulk material and the biological environment, which plays a key role in the optimized performance of medical devices. In this work, we report a new superhydrophilic adsorbate, called L-cysteine betaine (Cys-b), having branched zwitterionic groups that give rise to surfaces and nanoparticles with enhanced chemical stability, biofouling resistance, and inertness to environmental changes. Cys-b was synthesized from the amphoteric sulfur-containing amino acid, L-cysteine (Cys), by quaternization of its amino group. Gold surfaces modified with Cys-b exhibited prominent repellence against the nonspecific adsorption of proteins, bacteria, and fibroblast cells. In addition, Cys-b existed in zwitterionic form over a wide pH range (i.e., pH 3.4 to 10.8), and showed excellent suppression in photoinduced oxidation on gold substrates. Furthermore, the modification of hollow Ag@Au nanoshells with Cys-b gave rise to nanoparticles with excellent colloidal stability and resistance to coordinative interaction with Cu(2+). Taken together, the unique features of Cys-b offer a new nanoscale coating for use in a wide spectrum of applications.

  2. All-atom molecular dynamics simulation of a photosystem I/detergent complex

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

    Harris, Bradley J.; Cheng, Xiaolin; Frymier, Paul

    2014-09-18

    All-atom molecular dynamics (MD) simulation was used to investigate the solution structure and dynamics of the photosynthetic pigment protein complex photosystem I (PSI) from Thermosynechococcus elongatus embedded in a toroidal belt of n-dodecyl-β-d-maltoside (DDM) detergent. Evaluation of root-mean-square deviations (RMSDs) relative to the known crystal structure show that the protein complex surrounded by DDM molecules is stable during the 200 ns simulation time, and root-mean-square fluctuation (RMSF) analysis indicates that regions of high local mobility correspond to solvent-exposed regions such as turns in the transmembrane α-helices and flexible loops on the stromal and lumenal faces. Comparing the protein detergent complexmore » to a pure detergent micelle, the detergent surrounding the PSI trimer is found to be less densely packed but with more ordered detergent tails, contrary to what is seen in most lipid bilayer models. We also investigated any functional implications for the observed conformational dynamics and protein detergent interactions, discovering interesting structural changes in the psaL subunits associated with maintaining the trimeric structure of the protein. Moreover, we find that the docking of soluble electron mediators such as cytochrome c 6 and ferredoxin to PSI is not significantly impacted by the solubilization of PSI in detergent.« less

  3. All-atom molecular dynamics simulation of a photosystem i/detergent complex.

    PubMed

    Harris, Bradley J; Cheng, Xiaolin; Frymier, Paul

    2014-10-09

    All-atom molecular dynamics (MD) simulation was used to investigate the solution structure and dynamics of the photosynthetic pigment-protein complex photosystem I (PSI) from Thermosynechococcus elongatus embedded in a toroidal belt of n-dodecyl-β-d-maltoside (DDM) detergent. Evaluation of root-mean-square deviations (RMSDs) relative to the known crystal structure show that the protein complex surrounded by DDM molecules is stable during the 200 ns simulation time, and root-mean-square fluctuation (RMSF) analysis indicates that regions of high local mobility correspond to solvent-exposed regions such as turns in the transmembrane α-helices and flexible loops on the stromal and lumenal faces. Comparing the protein-detergent complex to a pure detergent micelle, the detergent surrounding the PSI trimer is found to be less densely packed but with more ordered detergent tails, contrary to what is seen in most lipid bilayer models. We also investigated any functional implications for the observed conformational dynamics and protein-detergent interactions, discovering interesting structural changes in the psaL subunits associated with maintaining the trimeric structure of the protein. Importantly, we find that the docking of soluble electron mediators such as cytochrome c6 and ferredoxin to PSI is not significantly impacted by the solubilization of PSI in detergent.

  4. Preservatives and fragrances in selected consumer-available cosmetics and detergents.

    PubMed

    Yazar, Kerem; Johnsson, Stina; Lind, Marie-Louise; Boman, Anders; Lidén, Carola

    2011-05-01

    Preservatives and fragrances are important and frequent skin sensitizers, found in a wide range of products intended for personal and occupational use. To examine the use of preservatives and fragrances in certain cosmetics and detergents on the market. The product types studied were shampoos, hair conditioners, liquid soaps, wet tissues, washing-up liquids, and multi-purpose cleaners. Ingredient labels of 204 cosmetic products and ingredient data sheets of 97 detergents, available on company websites, were examined. The preservatives most frequently identified were phenoxyethanol, methylparaben, sodium benzoate, propylparaben, and methylchloroisothiazolinone/methylisothiazolinone. Parabens were found in 44% of cosmetics and 9% of detergents; formaldehyde-releasers in 25% of cosmetics and 8% of detergents; and isothiazolinones in 23% of cosmetics and 28% of detergents. The fragrances most frequently identified were linalool, limonene, hexyl cinnamal, butylphenyl methylpropional, and citronellol. Eighty-eight per cent of the products contained fragrances, and any of the 26 fragrances requiring labelling were found in half of the cosmetics and one-third of the detergents. Several preservatives and fragrances with well-known skin-sensitizing potential were common in the examined product types. Such products may be used several times a day by consumers and workers. © 2010 John Wiley & Sons A/S.

  5. METHOD FOR THE MEASUREMENT OF SITE-SPECIFIC TAUTOMERIC AND ZWITTERIONIC MICROSPECIES EQUILIBRIUM CONSTANTS

    EPA Science Inventory

    We describe a method for the individual measurement of simultaneously occurring, unimolecular, site-specific “microequilibrium” constants as in, for example, prototropic tautomerism and zwitterionic equilibria. Our method represents an elaboration of that of Nygren et al. (Anal. ...

  6. The influence of detergents and active components of detergent on bioproduction of organic matters and enzymatic activity of some species of fungi.

    PubMed

    Stojanović, Jelica; Stojanović, Marina; Iles, Deana; Mijusković, Zoran

    2004-01-01

    Detergent (Merix, "Merima " Krusevac) applied in concentration of 1% vol. showed specific influence on the bioproduction of some 15 different amino acids and on the enzyme activity of the species of fungi A. niger, A. alternata and T. roseum. Detergent has significantly stimulated the production of 15 analyzed amino acids of the fungi species A. niger. The same applied concentration of detergent has decreased or considerably decreased the production of some 14 of totally 15 analyzed amino acids of investigated fungi species A. alternata and T. roseum. The enzyme activity of the fungi A. niger was more intensive in relation to the species A. alternata and T. roseum during the experimental period or in some phases of the experimental period. The detergent component, ethoxyled oleyl-cetyl alcohol, in concentration of 0.01%, 0.1% and 1% showed an inhibitory effect, or significant inhibitory effect on the enzyme activity of the examined species of fungi (A. niger, A. alternata and T. roseum).

  7. Cellular localization and detergent dependent oligomerization of rice allene oxide synthase-1.

    PubMed

    Yoeun, Sereyvath; Kim, Jeong-Il; Han, Oksoo

    2015-01-01

    Allene oxide synthase-1 from Oryza sativa (OsAOS1) localizes to the chloroplast, but lacks a putative chloroplast targeting sequence typically found in dicot AOS. Here, kinetic parameters and the oligomerization state/subunit composition of OsAOS1 were characterized in vitro in the absence or presence of detergent micelles. The catalytic efficiency (k(cat)/K(m)) of OsAOS1 reached a maximum near the critical micelle concentration for polyoxyethylene 10 tridecyl ether. Native gel analysis showed that OsAOS1 exists as a multimer in the absence of detergent micelles. The multimeric form of OsAOS1 was stably cross-linked in the absence of detergents, while only monomeric OsAOS1 was detected in the presence of detergent micelles. Gel filtration analysis indicated that the oligomeric state of OsAOS1 depends strongly on the detergents and that the monomer becomes the predominant form in the presence of detergent micelles. These data suggest that the detergent-dependent oligomeric state of OsAOS1 is an important factor for the regulation of its catalytic efficiency.

  8. A robust method to screen detergents for membrane protein stabilization, revisited.

    PubMed

    Champeil, Philippe; Orlowski, Stéphane; Babin, Simon; Lund, Sten; le Maire, Marc; Møller, Jesper; Lenoir, Guillaume; Montigny, Cédric

    2016-10-15

    This report is a follow up of our previous paper (Lund, Orlowski, de Foresta, Champeil, le Maire and Møller (1989), J Biol Chem 264:4907-4915) showing that solubilization in detergent of a membrane protein may interfere with its long-term stability, and proposing a protocol to reveal the kinetics of such irreversible inactivation. We here clarify the fact that when various detergents are tested for their effects, special attention has of course to be paid to their critical micelle concentration. We also investigate the effects of a few more detergents, some of which have been recently advertised in the literature, and emphasize the role of lipids together with detergents. Among these detergents, lauryl maltose neopentyl glycol (LMNG) exerts a remarkable ability, even higher than that of β-dodecylmaltoside (DDM), to protect our test enzyme, the paradigmatic P-type ATPase SERCA1a from sarcoplasmic reticulum. Performing such experiments for one's favourite protein probably remains useful in pre-screening assays testing various detergents. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. Synchronized Regulation of Different Zwitterionic Metabolites in the Osmoadaption of Phytoplankton

    PubMed Central

    Gebser, Björn; Pohnert, Georg

    2013-01-01

    The ability to adapt to different seawater salinities is essential for cosmopolitan marine phytoplankton living in very diverse habitats. In this study, we examined the role of small zwitterionic metabolites in the osmoadaption of two common microalgae species Emiliania huxleyi and Prorocentrum minimum. By cultivation of the algae under salinities between 16‰ and 38‰ and subsequent analysis of dimethylsulfoniopropionate (DMSP), glycine betaine (GBT), gonyol, homarine, trigonelline, dimethylsulfonioacetate, trimethylammonium propionate, and trimethylammonium butyrate using HPLC-MS, we could reveal two fundamentally different osmoadaption mechanisms. While E. huxleyi responded with cell size reduction and a nearly constant ratio between the major metabolites DMSP, GBT and homarine to increasing salinity, osmolyte composition of P. minimum changed dramatically. In this alga DMSP concentration remained nearly constant at 18.6 mM between 20‰ and 32‰ but the amount of GBT and dimethylsulfonioacetate increased from 4% to 30% of total investigated osmolytes. Direct quantification of zwitterionic metabolites via LC-MS is a powerful tool to unravel the complex osmoadaption and regulation mechanisms of marine phytoplankton. PMID:23774888

  10. Reaction of iminopropadienones with amines--formation of zwitterionic intermediates, ketenes, and ketenimines.

    PubMed

    Veedu, Rakesh Naduvile; Kokas, Okanya J; Couturier-Tamburelli, Isabelle; Koch, Rainer; Aycard, Jean-Pierre; Borget, Fabien; Wentrup, Curt

    2008-10-09

    Five aryliminopropadienones 4a- d have been synthesized by flash vacuum thermolysis (FVT) by using two different precursors in each case. These compounds were deposited at 50 K at a pressure of ca. 10(-6) mbar together with three different nucleophiles, namely, trimethylamine (TMA), dimethylamine (DMA), and diethylamine (DEA), in order to study their reactions as neat solids during warm-up by FTIR spectroscopy. The reaction with TMA showed that a zwitterionic species (5 and/or 6) was formed in all the cases. With DMA and DEA, an alpha-oxoketenimine and/or an imidoylketene (7 and 8 or 9 and 10) was formed as the final product. In addition, several bands were observed, which can be assigned to zwitterionic intermediates (11 or 12). Optimized structures and vibrational spectra for all products were calculated at the B3LYP/6-31G(d) level of theory by using the polarizable continuum model (epsilon = 5).

  11. Light-Induced Gelling in a Micellar Fluid Based on a Zwitterionic Surfactant.

    NASA Astrophysics Data System (ADS)

    Kumar, Rakesh; Raghavan, Srinivasa

    2007-03-01

    Fluids with photoresponsive rheological properties (i.e. photorheological or PR fluids) can be useful in a range of applications, such as in dampers, sensors, and valves for microfluidic or MEMS devices. Previously, we have demonstrated a cationic surfactant-based PR fluid whose viscosity can be rapidly decreased by UV irradiation. This viscosity decrease was not reversible. Here, we describe a different formulation based on a zwitterionic surfactant that shows a rapid increase in viscosity (gelling) upon exposure to UV radiation. The formulation consists of the zwitterionic surfactant and a photosensitive cinnamic acid derivative. Initially, the viscosity of the fluid is low indicating the presence of small micelles. Upon UV irradiation, the cinnamic acid derivative is photoisomerized from trans to cis. In turn, the small micelles transform into long wormlike micelles, thus increasing the solution viscosity by more than five orders of magnitude. Small angle neutron scattering (SANS) data confirms the dramatic increase in micelle length. Possible reasons for such changes in micelle dimensions will be discussed.

  12. Controllable self-assembly of sodium caseinate with a zwitterionic vitamin-derived bolaamphiphile.

    PubMed

    Sun, Li-Hui; Sun, Yu-Long; Yang, Li-Jun; Zhang, Jian; Chen, Zhong-Xiu

    2013-11-06

    The control of self-assembly of sodium caseinate (SC) including the formation of mixed layers, microspheres, or nanoparticles is highly relevant to the microstructure of food and the design of promising drug delivery systems. In this paper, we designed a structure-switchable zwitterionic bolaamphiphile, 1,12-diaminododecanediorotate (DDO), from orotic acid, which has special binding sites and can guide the self-assembly of SC. Complexation between SC and DDO was investigated using dynamic light scattering, transmission electron microscopy, differential scanning calorimetry, and fluorescence spectra measurements. Monomeric DDO was bound to the negatively charged sites on the SC micelle and made the structure of SC more compact with decreased electrostatic repulsion between the head groups. Vesicular DDO led to reassociation of vesicles with enlarged size via preferable hydrophobic interactions. Moreover, the aggregation between SC and DDO was found to be temperature-dependent and reversible. This research provides an effective way to control the reversible self-assembly of SC by the zwitterionic vitamin-derived bolaamphiphile.

  13. Mechanisms of anionic detergent-induced hemolysis.

    PubMed

    Chernitsky, E; Senkovich, O

    1998-09-01

    The effect of osmotic protectors (sucrose and polyethylene glycols) and of a decrease in the detergent concentration at different points of hemolysis of human erythrocytes by sodium dodecyl sulphate on the shape of kinetic curves of hemolysis were studied. It is shown that slow detergent-induced hemolysis follows the colloid-osmotic mechanisms. Evidence is provided that rapid hemolysis by sodium dodecyl sulphate is caused by opening of large pores sufficient for the release of hemoglobin molecules rather than by the colloid-osmotic mechanism, and that the kinetics of hemolysis is mainly determined by time dependence of the opening probability of these pores.

  14. Availability of manufacturers' information on efficacy and compatibility of detergents used for cleaning dental instruments.

    PubMed

    Calvert, G; Murray, C A; Smith, A J; Hurrell, D

    2012-05-25

    To review physico-chemical data supplied for commercially available detergents marketed for manual and/or ultrasonic cleansing of reusable dental instruments. Manufacturers/suppliers of commercially available detergents for manual or ultrasonic cleaning of dental instruments within primary dental care were invited to supply product information. A structured questionnaire requested details on a range of physical and chemical properties for each detergent. Seventeen detergent manufacturers/suppliers, encompassing 31 commercially available detergents were identified. Ten of the 17 manufacturers provided information on 23 (74%) of the detergent formulations. Nine detergents were of neutral pH, ten mild alkalis (pH 7.5-10.5) and four strong alkalis (pH >10.5). Sixteen detergents were recommended for ultrasonic and manual cleaning, four stated ultrasonic use and three manual only. Ten detergents cited enzymatic activity as their main mode of action, but only six manufacturers provided detailed information. Four detergents recommended by manufacturers as suitable for manual washing had a strong alkaline pH (>10.5), presenting chemical hazards to users. Two strong alkaline detergents did not warn users of potential adverse effects of such alkaline solutions (corrosion) upon aluminium containing instruments. Only one detergent had investigated the potential toxicity of detergent residuals remaining on instruments after reprocessing. It has proven challenging to collate physico-chemical data on detergents suitable for use in manual and/or ultrasonic cleaning of dental instruments in general dental practice. Standardisation of information on the nature and efficacy of dental detergents in a readily accessible form would be beneficial to dental practice.

  15. Zwitterion radicals and anion radicals from electron transfer and solvent condensation with the fingerprint developing agent ninhydrin.

    PubMed

    Schertz, T D; Reiter, R C; Stevenson, C D

    2001-11-16

    Ninhydrin (the fingerprint developing agent) spontaneously dehydrates in liquid ammonia and in hexamethylphosphoramide (HMPA) to form indantrione, which has a sufficiently large solution electron affinity to extract an electron from the solvent (HMPA) to produce the indantrione anion radical. In liquid NH(3), the presence of trace amounts of amide ion causes the spontaneous formation of an anion radical condensation product, wherein the no. 2 carbon (originally a carbonyl carbon) becomes substituted with -NH(2) and -OH groups. In HMPA, the indantrione anion radical spontaneously forms condensation products with the HMPA to produce a variety of zwitterionic radicals, wherein the no. 2 carbon becomes directly attached to a nitrogen of the HMPA. The mechanisms for the formation of the zwitterionic paramagnetic condensation products are analogous to that observed in the reaction of ninhydrin with amino acids to yield Ruhemann's Purple, the contrast product in fingerprint development. The formation of anion and zwitterionic radical condensation products from ninhydrin and nitrogen-containing solvents may represent an example of a host of analogous polyketone-solvent reactions.

  16. Using temperature-responsive zwitterionic surfactant to enhance the enzymatic hydrolysis of lignocelluloses and recover cellulase by cooling.

    PubMed

    Cai, Cheng; Pang, Yuxia; Zhan, Xuejuan; Zeng, Meijun; Lou, Hongming; Qian, Yong; Yang, Dongjie; Qiu, Xueqing

    2017-11-01

    Some zwitterionic surfactants exhibit upper critical solution temperature (UCST) in aqueous solutions. For the zwitterionic surfactant solution mixed with cellulase, when its temperature is below UCST, the cellulase can be recovered by coprecipitation with zwitterionic surfactant. In this work, 3-(Hexadecyldimethylammonio) propanesulfonate (SB3-16) was selected to enhance the enzymatic hydrolysis of lignocelluloses and recover the cellulase. After adding 2mmol/L of SB3-16, the enzymatic digestibility of eucalyptus pretreated by dilute acid (Eu-DA) and by sulfite (Eu-SPORL) increased from 27.9% and 35.1% to 72.6% and 89.7%, respectively. The results showed that SB3-16 could reduce the non-productive adsorption of cellulase on hydrophobic interface, while it did not significantly inhibit the activity of cellulase. For the solution contained 1wt% SB3-16 and 200mg protein/L CTec2 cellulase, 55.2% of protein could be recovered by cooling. The filter paper activity of the recovered cellulase was 1.93FPU/mg protein, which was 95.8% of its initial activity. Copyright © 2017. Published by Elsevier Ltd.

  17. Monovalent Ions and Water Dipoles in Contact with Dipolar Zwitterionic Lipid Headgroups-Theory and MD Simulations

    PubMed Central

    Velikonja, Aljaž; Perutkova, Šarka; Gongadze, Ekaterina; Kramar, Peter; Polak, Andraž; Maček-Lebar, Alenka; Iglič, Aleš

    2013-01-01

    The lipid bilayer is a basic building block of biological membranes and can be pictured as a barrier separating two compartments filled with electrolyte solution. Artificial planar lipid bilayers are therefore commonly used as model systems to study the physical and electrical properties of the cell membranes in contact with electrolyte solution. Among them the glycerol-based polar phospholipids which have dipolar, but electrically neutral head groups, are most frequently used in formation of artificial lipid bilayers. In this work the electrical properties of the lipid layer composed of zwitterionic lipids with non-zero dipole moments are studied theoretically. In the model, the zwitterionic lipid bilayer is assumed to be in contact with aqueous solution of monovalent salt ions. The orientational ordering of water, resulting in spatial variation of permittivity, is explicitly taken into account. It is shown that due to saturation effect in orientational ordering of water dipoles the relative permittivity in the zwitterionic headgroup region is decreased, while the corresponding electric potential becomes strongly negative. Some of the predictions of the presented mean-field theoretical consideration are critically evaluated using the results of molecular dynamics (MD) simulation. PMID:23434651

  18. Detergents: Friends not foes for high-performance membrane proteomics toward precision medicine.

    PubMed

    Zhang, Xi

    2017-02-01

    Precision medicine, particularly therapeutics, emphasizes the atomic-precise, dynamic, and systems visualization of human membrane proteins and their endogenous modifiers. For years, bottom-up proteomics has grappled with removing and avoiding detergents, yet faltered at the therapeutic-pivotal membrane proteins, which have been tackled by classical approaches and are known for decades refractory to single-phase aqueous or organic denaturants. Hydrophobicity and aggregation commonly challenge tissue and cell lysates, biofluids, and enriched samples. Frequently, expected membrane proteins and peptides are not identified by shotgun bottom-up proteomics, let alone robust quantitation. This review argues the cause of this proteomic crisis is not detergents per se, but the choice of detergents. Recently, inclusion of compatible detergents for membrane protein extraction and digestion has revealed stark improvements in both quantitative and structural proteomics. This review analyzes detergent properties behind recent proteomic advances, and proposes that rational use of detergents may reconcile outstanding membrane proteomics dilemmas, enabling ultradeep coverage and minimal artifacts for robust protein and endogenous PTM measurements. The simplicity of detergent tools confers bottom-up membrane proteomics the sophistication toward precision medicine. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Quantitative fabrication, performance optimization and comparison of PEG and zwitterionic polymer antifouling coatings.

    PubMed

    Xing, Cheng-Mei; Meng, Fan-Ning; Quan, Miao; Ding, Kai; Dang, Yuan; Gong, Yong-Kuan

    2017-09-01

    A versatile fabrication and performance optimization strategy of PEG and zwitterionic polymer coatings is developed on the sensor chip of surface plasma resonance (SPR) instrument. A random copolymer bearing phosphorylcholine zwitterion and active ester side chains (PMEN) and carboxylic PEG coatings with comparable thicknesses were deposited on SPR sensor chips via amidation coupling on the precoated polydopamine (PDA) intermediate layer. The PMEN coating showed much stronger resistance to bovine serum albumin (BSA) adsorption than PEG coating at very thin thickness (∼1nm). However, the BSA resistant efficacy of PEG coating could exceed that of PMEN due to stronger steric repelling effect when the thickness increased to 1.5∼3.3nm. Interestingly, both the PEG and PMEN thick coatings (≈3.6nm) showed ultralow fouling by BSA and bovine plasma fibrinogen (Fg). Moreover, changes in the PEG end group from -OH to -COOH, protein adsorption amount could increase by 10-fold. Importantly, the optimized PMEN and PEG-OH coatings were easily duplicated on other substrates due to universal adhesion of the PDA layer, showed excellent resistance to platelet, bacteria and proteins, and no significant difference in the antifouling performances was observed. These detailed results can explain the reported discrepancy in performances between PEG and zwitterionic polymer coatings by thickness. This facile and substrate-independent coating strategy may benefit the design and manufacture of advanced antifouling biomedical devices and long circulating nanocarriers. Prevention of biofouling is one of the biggest challenges for all biomedical applications. However, it is very difficult to fabricate a highly hydrophilic antifouling coating on inert materials or large devices. In this study, PEG and zwitterion polymers, the most widely investigated polymers with best antifouling performance, are conveniently immobilized on different kinds of substrates from their aqueous solutions by

  20. Harmful Effects of Synthetic Surface-Active Detergents against Atopic Dermatitis.

    PubMed

    Deguchi, Hajime; Aoyama, Riho; Takahashi, Hideaki; Isobe, Yoshinari; Tsutsumi, Yutaka

    2015-01-01

    We report herein two cases of intractable atopic dermatitis successfully treated by simply avoiding the contact with surface-active detergents in the daily life and living. The detergents were closely related to the exacerbation and remission of the disease. Steroid ointment was no longer used. We discuss that the removal of horny layer lipids by surface-active detergents accelerates the transepidermal water loss and disturbs the barrier function of the epidermis and thus is intimately involved in the pathogenesis of atopic dermatitis.

  1. Ecotoxicity evaluation of a liquid detergent using the automatic biotest ECOTOX.

    PubMed

    Azizullah, Azizullah; Richter, Peter; Ullah, Waheed; Ali, Imran; Häder, Donat-Peter

    2013-08-01

    Synthetic detergents are common pollutants reaching aquatic environments in different ways after usage at homes, institutions and industries. In this study a liquid detergent, used for dish washing, was evaluated for its toxicity during long- and short-term tests using the automatic biotest ECOTOX. Different parameters of Euglena gracilis like motility, swimming velocity, gravitactic orientation, cell compactness and cell growth were used as end points. In short-term experiments, the maximum adverse effects on motility, velocity, cell shape and gravitaxis were observed after 1 h of exposure. With further increase in exposure time to the detergent a slight recovery of these parameters was observed. In long-term experiments, the detergent caused severe disturbances to E. gracilis. Motility, cell growth and cell compactness (shape) with EC50 values of 0.064, 0.18 and 2.05 %, respectively, were found as the most sensitive parameters to detergent stress. There was a slight positive effect on gravitactic orientation at the lowest two concentrations; at higher concentrations of the detergent cells orientation was highly impaired giving EC50 values of 1.75 and 2.52 % for upward swimming and r-value, respectively.

  2. Stakeholders' Perspectives towards the Use of the Comprehensive Health Assessment Program (CHAP) for Adults with Intellectual Disabilities in Manitoba

    ERIC Educational Resources Information Center

    Shooshtari, Shahin; Temple, Beverley; Waldman, Celeste; Abraham, Sneha; Ouellette-Kuntz, Héléne; Lennox, Nicholas

    2017-01-01

    Background: No standardized tool is used in Canada for comprehensive health assessments of adults with intellectual disabilities. This study was conducted to determine the feasibility of implementing the Comprehensive Health Assessment Program (CHAP) in Manitoba, Canada. Method: This was a qualitative study using a purposive sample of physicians,…

  3. A METHOD FOR THE MEASUREMENT OF SITE-SPECIFIC TAUTOMERIC AND ZWITTERIONIC MICROSPECIES EQUILIBRIUM CONSTANTS

    EPA Science Inventory

    We describe a method for the individual measurement of simultaneously occurring, unimolecular, site-specific "microequilibrium" constants as in, for example, prototropic tautomerism and zwitterionic equilibria. Our method represents an elaboration of that of Nygren et al. (Anal. ...

  4. Mixing and Matching Detergents for Membrane Protein NMR Structure Determination

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

    Columbus, Linda; Lipfert, Jan; Jambunathan, Kalyani

    2009-10-21

    One major obstacle to membrane protein structure determination is the selection of a detergent micelle that mimics the native lipid bilayer. Currently, detergents are selected by exhaustive screening because the effects of protein-detergent interactions on protein structure are poorly understood. In this study, the structure and dynamics of an integral membrane protein in different detergents is investigated by nuclear magnetic resonance (NMR) and electron paramagnetic resonance (EPR) spectroscopy and small-angle X-ray scattering (SAXS). The results suggest that matching of the micelle dimensions to the protein's hydrophobic surface avoids exchange processes that reduce the completeness of the NMR observations. Based onmore » these dimensions, several mixed micelles were designed that improved the completeness of NMR observations. These findings provide a basis for the rational design of mixed micelles that may advance membrane protein structure determination by NMR.« less

  5. Impact of detergent systems on bacterial survival on laundered fabrics.

    PubMed Central

    Jaska, J M; Fredell, D L

    1980-01-01

    The survival of Staphylococcus aureus was determined from inoculated swatches laundered in either a phosphate or a phosphate-substitute detergent. In a Plackett-Burman design study, the independent variables of detergent type, concentration, and variation, wash water temperature, soil load, cycle time, and water hardness were assigned high and low values. Wash water temperatures of 27, 38, 49, and 60 degrees C were employed. Viable bacteria were recovered from macerated swatches. Statistical analysis disclosed that there was no practical difference in the ability of phosphate or phosphate-substitute detergents to reduce the level of S. aureus on the laundered swatches in this controlled design. Analysis did reveal that water temperature was the most significant independent variables. The remaining variables did not appear to have any practical significance upon bacterial reduction. This bacteriological study did not evaluate other essential detergent properties. PMID:7377775

  6. Murein Lytic Enzyme TgaA of Bifidobacterium bifidum MIMBb75 Modulates Dendritic Cell Maturation through Its Cysteine- and Histidine-Dependent Amidohydrolase/Peptidase (CHAP) Amidase Domain

    PubMed Central

    Zanoni, Ivan; Balzaretti, Silvia; Miriani, Matteo; Taverniti, Valentina; De Noni, Ivano; Presti, Ilaria; Stuknyte, Milda; Scarafoni, Alessio; Arioli, Stefania; Iametti, Stefania; Bonomi, Francesco; Mora, Diego; Karp, Matti; Granucci, Francesca

    2014-01-01

    Bifidobacteria are Gram-positive inhabitants of the human gastrointestinal tract that have evolved close interaction with their host and especially with the host's immune system. The molecular mechanisms underlying such interactions, however, are largely unidentified. In this study, we investigated the immunomodulatory potential of Bifidobacterium bifidum MIMBb75, a bacterium of human intestinal origin commercially used as a probiotic. Particularly, we focused our attention on TgaA, a protein expressed on the outer surface of MIMBb75's cells and homologous to other known bacterial immunoactive proteins. TgaA is a peptidoglycan lytic enzyme containing two active domains: lytic murein transglycosylase (LT) and cysteine- and histidine-dependent amidohydrolase/peptidase (CHAP). We ran immunological experiments stimulating dendritic cells (DCs) with the B. bifidum MIMBb75 and TgaA, with the result that both the bacterium and the protein activated DCs and triggered interleukin-2 (IL-2) production. In addition, we observed that the heterologous expression of TgaA in Bifidobacterium longum transferred to the bacterium the ability to induce IL-2. Subsequently, immunological experiments performed using two purified recombinant proteins corresponding to the single domains LT and CHAP demonstrated that the CHAP domain is the immune-reactive region of TgaA. Finally, we also showed that TgaA-dependent activation of DCs requires the protein CD14, marginally involves TRIF, and is independent of Toll-like receptor 4 (TLR4) and MyD88. In conclusion, our study suggests that the bacterial CHAP domain is a novel microbe-associated molecular pattern actively participating in the cross talk mechanisms between bifidobacteria and the host's immune system. PMID:24814791

  7. Chromatographic behavior of zwitterionic enalapril-exploring the conditions for lipophilicity assessment.

    PubMed

    Gikas, Spyros; Tsopelas, Fotios; Giaginis, Costas; Dimitrakopoulos, John; Livadara, Theodora; Archontaki, Helen; Tsantili-Kakoulidou, Anna

    2008-11-04

    The chromatographic behavior of enalapril was investigated under different stationary and mobile phase conditions in an effort to unravel interferences in the underlying retention mechanism, which would affect its relation to octanol-water partitioning. Extrapolated retention factors, logk(w), were used as relevant chromatographic indices. The retention/pH profile was established and the peak split phenomenon, associated with cis/trans interconversion, was also monitored as a function of pH. The pH at maximum retention and minimum peak split occurrence was chosen for further investigation, so that the presence of zwitterionic structure was guaranteed and any effect of cis/trans interconversion could be ignored. Retention of zwitterionic enalapril was found to be very sensitive to mobile phase conditions in regard to organic modifier as well to the aqueous component. The use of morpholine-propanesulfonic acid (MOPS) as buffer and the presence of n-octanol as mobile phase additive proved critical factors for maximum suppression of secondary interactions. Nevertheless, the corresponding extrapolated retention factor was considerably larger than octanol-water logD value at the isoelectric point. However, logk(w) could be successfully converted to logD by means of a calibration equation established for ionized acidic compounds.

  8. 40 CFR 417.160 - Applicability; description of the manufacture of liquid detergents subcategory.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... manufacture of liquid detergents subcategory. 417.160 Section 417.160 Protection of Environment ENVIRONMENTAL... CATEGORY Manufacture of Liquid Detergents Subcategory § 417.160 Applicability; description of the manufacture of liquid detergents subcategory. The provisions of this subpart are applicable to discharges...

  9. 76 FR 9013 - Agency Information Collection Activities; Proposed Collection; Comment Request; Detergent Gasoline

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-16

    ... Activities; Proposed Collection; Comment Request; Detergent Gasoline AGENCY: Environmental Protection Agency... this action are those who (1) Manufacture gasoline, post-refinery component, or detergent additives, (2) blend detergent additives into gasoline or post-refinery component, or (3) transport or receive a...

  10. Effect of synthetic detergents on germination of fern spores

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

    Devi, Y.; Devi, S.

    Synthetic detergents constitute one of the most important water pollutants by contaminating the lakes and rivers through domestic and industrial use. Considerable information is now available for the adverse effects of detergents an aquatic fauna including fish, algae, and higher aquatic plants. Marked inhibition of germination in orchids and brinjals and of seedlings growth in raddish suggest that rapidly growing systems could be sensitive to detergent polluted water. The present study of the effect of linear alkyl benzene sulphonate on germination of the spores of a fern, Diplazium esculentum aims at the understanding of the effects of water pollution onmore » pteridophytes and the development of spore germination assay for phytoxicity evaluation.« less

  11. Use of anionic denaturing detergents to purify insoluble proteins after overexpression

    PubMed Central

    2012-01-01

    Background Many proteins form insoluble protein aggregates, called “inclusion bodies”, when overexpressed in E. coli. This is the biggest obstacle in biotechnology. Ever since the reversible denaturation of proteins by chaotropic agents such as urea or guanidinium hydrochloride had been shown, these compounds were predominantly used to dissolve inclusion bodies. Other denaturants exist but have received much less attention in protein purification. While the anionic, denaturing detergent sodiumdodecylsulphate (SDS) is used extensively in analytical SDS-PAGE, it has rarely been used in preparative purification. Results Here we present a simple and versatile method to purify insoluble, hexahistidine-tagged proteins under denaturing conditions. It is based on dissolution of overexpressing bacterial cells in a buffer containing sodiumdodecylsulfate (SDS) and whole-lysate denaturation of proteins. The excess of detergent is removed by cooling and centrifugation prior to affinity purification. Host- and overexpressed proteins do not co-precipitate with SDS and the residual concentration of detergent is compatible with affinity purification on Ni/NTA resin. We show that SDS can be replaced with another ionic detergent, Sarkosyl, during purification. Key advantages over denaturing purification in urea or guanidinium are speed, ease of use, low cost of denaturant and the compatibility of buffers with automated FPLC. Conclusion Ionic, denaturing detergents are useful in breaking the solubility barrier, a major obstacle in biotechnology. The method we present yields detergent-denatured protein. Methods to refold proteins from a detergent denatured state are known and therefore we propose that the procedure presented herein will be of general application in biotechnology. PMID:23231964

  12. Determination and residual characteristic of alkylphenols in household food detergents of Taiwan.

    PubMed

    Pan, Yi-Ping; Tsai, Shih-Wei

    2009-07-01

    The non-ionic surfactants are mostly composed of alkylphenols for the ingredients of synthetic food detergents. Due to the ability to mimic hormones, it has been noticed that the exposures of alkylphenols might cause a variety of adverse effects. To assess the associate risks from possible exposures, concentrations of alkylphenols, including 4-nonylphenol (4-NP), technical nonylphenol isomers (t-NP(S)), and 4-tert-octylphenol (4-t-OP), in household food detergents of Taiwan were determined. Gas chromatography with mass spectrometer (GC/MS) was used to analyze alkylphenols in samples. The Taguchi experimental design was utilized to study the possible factors that might affect the residual characteristics of alkylphenols from detergents on dishware and fruits. By the analysis of variance, the orders of importance of different parameters were determined. The results showed that the concentrations of alkylphenols in food detergents ranged from 1.71 x 10(-5) to 2.13 x 10(-3) (APs/detergent, mgg(-1)). For residual characteristics, the cleaning temperature was found to be the only significant factor that will affect the 4-t-OP left on the dishware, while the concentrations of detergents used will affect the left of t-NPs and 4-NP on dishware as well. On the other hand, the varieties of fruits, the concentrations of detergents, and the concentrations of alkylphenols were found to have significant effects for the t-NPs left on fruits. As for the exposure assessments, the maximum dose of APs exposures from the use of household food detergents in Taiwan was also estimated in the study.

  13. Chronic toxicity of a laundry detergent to the freshwater flagellate Euglena gracilis.

    PubMed

    Azizullah, Azizullah; Richter, Peter; Jamil, Muhammad; Häder, Donat-Peter

    2012-10-01

    Chronic toxicity of the common laundry detergent Ariel on the freshwater alga Euglena gracilis was investigated by growing the alga in a medium containing the detergent for 7 days. Cell density, motility, swimming velocity, gravitactic orientation, cell shape, photosynthesis and concentration of light-harvesting pigments were used as end point parameters for the assessment of toxicity. Cell density was significantly reduced at a concentration of 1 mg l(-1) or above. Among the other tested parameters, with the exception of cell shape, gravitaxis and chlorophyll b, all were adversely affected by the detergent at concentrations exceeding 1 mg l(-1). It is concluded that long-term (7-days) exposure to the detergent caused significant toxicity to E. gracilis. Furthermore, long-term tests with E. gracilis can be used as sensitive indicator for the toxicity assessment of laundry detergents in aquatic environments.

  14. Mechanism of the inhibitory effect of zwitterionic drugs (levofloxacin and grepafloxacin) on carnitine transporter (OCTN2) in Caco-2 cells.

    PubMed

    Hirano, Takeshi; Yasuda, Satoru; Osaka, Yuki; Kobayashi, Masaki; Itagaki, Shirou; Iseki, Ken

    2006-11-01

    L-Carnitine plays an important role in lipid metabolism by facilitating the transport of long-chain fatty acids across the mitochondrial inner membrane followed by fatty acid beta-oxidation. It is known that L-carnitine exists as a zwitterion and that member of the OCTN family play an important role in its transport. The aims of this study were to characterize L-carnitine transport in the intestine by using Caco-2 cells and to elucidate the effects of levofloxacin (LVFX) and grepafloxacin (GPFX), which are zwitterionic drugs, on L-carnitine uptake. Kinetic analysis showed that the half-saturation Na+ concentration, Hill coefficient and Km value of L-carnitine uptake in Caco-2 cells were 10.3 +/- 4.5 mM, 1.09 and 8.0 +/- 1.0 microM, respectively, suggesting that OCTN2 mainly transports L-carnitine. LVFX and GPFX have two pKa values and the existence ratio of their zwitterionic forms is higher under a neutral condition than under an acidic condition. Experiments on the inhibitory effect of LVFX and GPFX on L-carnitine uptake showed that LVFX and GPFX inhibited L-carnitine uptake more strongly at pH 7.4 than at pH 5.5. It was concluded that the zwitterionic form of drugs plays an important role in inhibition of OCTN2 function.

  15. [Evaluation of the mutagenicity of detergents by tests on bacteria, plant cells and human leucocytes.].

    PubMed

    Feretti, Donatella; Pedrazzani, Roberta; Ceretti, Elisabetta; Zerbini, Ilaria; Gozio, Eleonora; Belotti, Caterina; Alias, Carlotta; Donato, Francesco; Gelatti, Umberto

    2009-01-01

    The aim of this study was to evaluate the mutagenicity of several traditional detergents and that of newer more biodegradable detergents, by using a bacterial test (Ames test), a plant cell test (Allium cepa micronuclei test) and a human leucocyte test (Comet test). All tests were conducted using a wide range of doses (1-2000 mg/l). None of the examined detergents induced mutations in S.typhimurium. One traditional detergent showed a genotoxic effect with the A. cepa test, while all newer detergents and one traditional detergent were shown by the Comet test to be capable of inducing DNA damage.

  16. A high-throughput differential filtration assay to screen and select detergents for membrane proteins

    PubMed Central

    Vergis, James M.; Purdy, Michael D.; Wiener, Michael C.

    2015-01-01

    Structural studies on integral membrane proteins are routinely performed on protein–detergent complexes (PDCs) consisting of purified protein solubilized in a particular detergent. Of all the membrane protein crystal structures solved to date, a subset of only four detergents has been used in more than half of these structures. Unfortunately, many membrane proteins are not well behaved in these four detergents and/or fail to yield well-diffracting crystals. Identification of detergents that maintain the solubility and stability of a membrane protein is a critical step and can be a lengthy and “protein-expensive” process. We have developed an assay that characterizes the stability and size of membrane proteins exchanged into a panel of 94 commercially available and chemically diverse detergents. This differential filtration assay (DFA), using a set of filtered microplates, requires sub-milligram quantities of purified protein and small quantities of detergents and other reagents and is performed in its entirety in several hours. PMID:20667442

  17. Recombinant Human Melatonin Receptor MT1 Isolated in Mixed Detergents Shows Pharmacology Similar to That in Mammalian Cell Membranes

    PubMed Central

    Logez, Christel; Berger, Sylvie; Legros, Céline; Banères, Jean-Louis; Cohen, William; Delagrange, Philippe; Nosjean, Olivier; Boutin, Jean A.; Ferry, Gilles

    2014-01-01

    The human melatonin MT1 receptor—belonging to the large family of G protein-coupled receptors (GPCRs)—plays a key role in circadian rhythm regulation and is notably involved in sleep disorders and depression. Structural and functional information at the molecular level are highly desired for fine characterization of this receptor; however, adequate techniques for isolating soluble MT1 material suitable for biochemical and biophysical studies remain lacking. Here we describe the evaluation of a panel of constructs and host systems for the production of recombinant human MT1 receptors, and the screening of different conditions for their solubilization and purification. Our findings resulted in the establishment of an original strategy using a mixture of Fos14 and CHAPS detergents to extract and purify a recombinant human MT1 from Pichia pastoris membranes. This procedure enabled the recovery of relatively pure, monomeric and ligand-binding active MT1 receptor in the near-milligram range. A comparative study based on extensive ligand-binding characterization highlighted a very close correlation between the pharmacological profiles of MT1 purified from yeast and the same receptor present in mammalian cell membranes. The high quality of the purified MT1 was further confirmed by its ability to activate its cognate Gαi protein partner when reconstituted in lipid discs, thus opening novel paths to investigate this receptor by biochemical and biophysical approaches. PMID:24959712

  18. Recombinant human melatonin receptor MT1 isolated in mixed detergents shows pharmacology similar to that in mammalian cell membranes.

    PubMed

    Logez, Christel; Berger, Sylvie; Legros, Céline; Banères, Jean-Louis; Cohen, William; Delagrange, Philippe; Nosjean, Olivier; Boutin, Jean A; Ferry, Gilles; Simonin, Frédéric; Wagner, Renaud

    2014-01-01

    The human melatonin MT1 receptor-belonging to the large family of G protein-coupled receptors (GPCRs)-plays a key role in circadian rhythm regulation and is notably involved in sleep disorders and depression. Structural and functional information at the molecular level are highly desired for fine characterization of this receptor; however, adequate techniques for isolating soluble MT1 material suitable for biochemical and biophysical studies remain lacking. Here we describe the evaluation of a panel of constructs and host systems for the production of recombinant human MT1 receptors, and the screening of different conditions for their solubilization and purification. Our findings resulted in the establishment of an original strategy using a mixture of Fos14 and CHAPS detergents to extract and purify a recombinant human MT1 from Pichia pastoris membranes. This procedure enabled the recovery of relatively pure, monomeric and ligand-binding active MT1 receptor in the near-milligram range. A comparative study based on extensive ligand-binding characterization highlighted a very close correlation between the pharmacological profiles of MT1 purified from yeast and the same receptor present in mammalian cell membranes. The high quality of the purified MT1 was further confirmed by its ability to activate its cognate Gαi protein partner when reconstituted in lipid discs, thus opening novel paths to investigate this receptor by biochemical and biophysical approaches.

  19. Detergent enhances binding of a secreted HLA-A2 molecule to solid phase peptides.

    PubMed

    Tussey, L G; Frelinger, J A

    1991-11-01

    We have constructed a secreted analogue (sA2) of the human class I molecule HLA-A2. sA2 was affinity purified both in the presence and absence of detergent and the effects of detergent on the magnitude and specificity of A2 binding to solid phase peptides tested. sA2 purified in the presence of detergent and detergent-solubilized A2 are shown to function comparably in the binding of the synthetic peptide M.Y + 57-68, a known T-cell epitope derived from the influenza A matrix protein. The molecules binding to M.Y + 57-68 typically represent 8% to 10% of the added protein. In contrast, less than 1% of sA2 protein purified in the absence of detergent binds M.Y + 57-68. This reduced binding is not due to a change in the affinity of sA2 for M.Y + 57-68. Addition of detergent at various stages of the purification and iodination procedures indicates that the longer the sA2 molecules are exposed to detergent the better they bind. However, the concentration of detergent during the actual binding assay does not appear to be critical. We also find that while the sA2-detergent and the sA2-no detergent molecules differ in the extent to which they bind various peptides, they do not differ in their patterns of binding. We conclude that detergent probably does not influence the specificity of class I/peptide binding but does increase the number of sA2 molecules that can participate in the binding of peptide either by generating and stabilizing "empty" sA2 molecules or by stabilizing a structure that is more amenable to binding peptide.

  20. Modification of Silicone Elastomer Surfaces with Zwitterionic Polymers: Short-Term Fouling Resistance and Triggered Biofouling Release.

    PubMed

    Shivapooja, Phanindhar; Yu, Qian; Orihuela, Beatriz; Mays, Robin; Rittschof, Daniel; Genzer, Jan; López, Gabriel P

    2015-11-25

    We present a method for dual-mode-management of biofouling by modifying surface of silicone elastomers with zwitterionic polymeric grafts. Poly(sulfobetaine methacrylate) was grafted from poly(vinylmethylsiloxane) elastomer substrates using thiol-ene click chemistry and surface-initiated, controlled radical polymerization. These surfaces exhibited both fouling resistance and triggered fouling-release functionality. The zwitterionic polymers exhibited fouling resistance over short-term (∼hours) exposure to bacteria and barnacle cyprids. The biofilms that eventually accumulated over prolonged-exposure (∼days) were easily detached by applying mechanical strain to the elastomer substrate. Such dual-functional surfaces may be useful in developing environmentally and biologically friendly coatings for biofouling management on marine, industrial, and biomedical equipment because they can obviate the use of toxic compounds.

  1. Development of Low Fouling and High Fouling-release Zwitterionic Marine Coatings

    DTIC Science & Technology

    2016-01-04

    amount, pigment , crosslinker type and catalyst amount. The details are shown below, 1. PDMS coating with zwitterionic copolymer incorporated. PDMS...onto aluminum panels. The coating can be cured within 3.5 hours. 2. PDMS coating with pigment incorporated. Various amount of the pigment cadmium...stirred vigorously until all the solid were dispersed uniformly. The obtained viscous liquid was then mixed with 3 g PDMS crosslinker, and applied onto

  2. [Response of the algae Gymnodinium kovalevskii (Dinophyta) to exposure to synthetic detergents and distillation].

    PubMed

    Aĭzdaĭcher, N A

    2000-01-01

    The effects of synthetic detergents and combined effects of synthetic detergents and water freshening on growth characteristics of the alga Gymnodinium kovalevskii (Dinophyta) were studied. Low concentrations of synthetic detergents (0.1 and 1.0 mg/l) stimulated the algal growth. Elevated concentrations inhibited cell division, affected their motility and induced morphological changes. Contamination with synthetic detergents adversely affected the adaptation plasticity of algae with respect to salinity.

  3. Detergent-free purification of ABC (ATP-binding-cassette) transporters.

    PubMed

    Gulati, Sonali; Jamshad, Mohammed; Knowles, Timothy J; Morrison, Kerrie A; Downing, Rebecca; Cant, Natasha; Collins, Richard; Koenderink, Jan B; Ford, Robert C; Overduin, Michael; Kerr, Ian D; Dafforn, Timothy R; Rothnie, Alice J

    2014-07-15

    ABC (ATP-binding-cassette) transporters carry out many vital functions and are involved in numerous diseases, but study of the structure and function of these proteins is often hampered by their large size and membrane location. Membrane protein purification usually utilizes detergents to solubilize the protein from the membrane, effectively removing it from its native lipid environment. Subsequently, lipids have to be added back and detergent removed to reconstitute the protein into a lipid bilayer. In the present study, we present the application of a new methodology for the extraction and purification of ABC transporters without the use of detergent, instead, using a copolymer, SMA (polystyrene-co-maleic acid). SMA inserts into a bilayer and assembles into discrete particles, essentially solubilizing the membrane into small discs of bilayer encircled by a polymer, termed SMALPs (SMA lipid particles). We show that this polymer can extract several eukaryotic ABC transporters, P-glycoprotein (ABCB1), MRP1 (multidrug-resistance protein 1; ABCC1), MRP4 (ABCC4), ABCG2 and CFTR (cystic fibrosis transmembrane conductance regulator; ABCC7), from a range of different expression systems. The SMALP-encapsulated ABC transporters can be purified by affinity chromatography, and are able to bind ligands comparably with those in native membranes or detergent micelles. A greater degree of purity and enhanced stability is seen compared with detergent solubilization. The present study demonstrates that eukaryotic ABC transporters can be extracted and purified without ever being removed from their lipid bilayer environment, opening up a wide range of possibilities for the future study of their structure and function.

  4. Corps Helicopter Attack Planning System (CHAPS). Positional Handbook. Appendix A. Messages. Appendix B. Statespace Construction Sample Session

    DTIC Science & Technology

    1989-10-01

    REVIEW MENU PROGRAM (S) CHAPS PURPOSE AND OVERVIEV The Do Review menu allows the user to select which missions to perform detailed analysis on and...input files must be resident on the computer you are running SUPR on. Any interface or file transfer programs must be successfully executed prior to... COMPUTER PROGRAM WAS DEVELOPED BY SYSTEMS CONTROL TECHNOLOGY FOR THE DEPUTY CHIEF OF STAFF/OPERATIONS,HQ USAFE. THE USE OF THE COMPUTER PROGRAM IS

  5. Summary of: availability of manufacturers' information on efficacy and compatibility of detergents used for cleaning dental instruments.

    PubMed

    Barker, Chris

    2012-05-25

    To review physico-chemical data supplied for commercially available detergents marketed for manual and/or ultrasonic cleansing of reusable dental instruments. Manufacturers/suppliers of commercially available detergents for manual or ultrasonic cleaning of dental instruments within primary dental care were invited to supply product information. A structured questionnaire requested details on a range of physical and chemical properties for each detergent. Seventeen detergent manufacturers/suppliers, encompassing 31 commercially available detergents were identified. Ten of the 17 manufacturers provided information on 23 (74%) of the detergent formulations. Nine detergents were of neutral pH, ten mild alkalis (pH 7.5-10.5) and four strong alkalis (pH >10.5). Sixteen detergents were recommended for ultrasonic and manual cleaning, four stated ultrasonic use and three manual only. Ten detergents cited enzymatic activity as their main mode of action, but only six manufacturers provided detailed information. Four detergents recommended by manufacturers as suitable for manual washing had a strong alkaline pH (>10.5), presenting chemical hazards to users. Two strong alkaline detergents did not warn users of potential adverse effects of such alkaline solutions (corrosion) upon aluminium containing instruments. Only one detergent had investigated the potential toxicity of detergent residuals remaining on instruments after reprocessing. It has proven challenging to collate physico-chemical data on detergents suitable for use in manual and/or ultrasonic cleaning of dental instruments in general dental practice. Standardisation of information on the nature and efficacy of dental detergents in a readily accessible form would be beneficial to dental practice.

  6. Detergent solubilization of the EGF receptor from A431 cells

    NASA Technical Reports Server (NTRS)

    Dayanidhi, R.; Rintoul, D. A.; Spooner, B. S. (Principal Investigator)

    1993-01-01

    Functional reconstitution of purified preparations of human epidermal growth factor receptor (EGFR) requires dissociation of the protein from its plasma membrane lipid environment. Solubilization of membrane proteins in this manner requires the use of detergents, which are known to disrupt plasma membrane lipid/protein interactions. We have investigated the ability of three nonionic detergents to solubilize the human EGFR selectively, and have also analyzed the effect of these various treatments on the intrinsic tyrosyl kinase activity of the receptor. The nonionic detergent known as n-octyl glucoside (n-octyl beta-D-glucopyranoside) was found to give the best combination of selectivity, yield, and maintenance of enzymatic activity of the human EGFR.

  7. Opposite behavior of two isozymes when refolding in the presence of non-ionic detergents.

    PubMed Central

    Doñate, F.; Artigues, A.; Iriarte, A.; Martinez-Carrion, M.

    1998-01-01

    GroEL has a greater affinity for the mitochondrial isozyme (mAAT) of aspartate aminotransferase than for its cytosolic counterpart (cAAT) (Mattingly JR Jr, Iriarte A, Martinez-Carrion M, 1995, J Biol Chem 270:1138-1148), two proteins that share a high degree of sequence similarity and an almost identical spatial structure. The effect of detergents on the refolding of these large, dimeric isozymes parallels this difference in behavior. The presence of non-ionic detergents such as Triton X-100 or lubrol at concentrations above their critical micelle concentration (CMC) interferes with reactivation of mAAT unfolded in guanidinium chloride but increases the yield of cAAT refolding at low temperatures. The inhibitory effect of detergents on the reactivation of mAAT decreases progressively as the addition of detergents is delayed after starting the refolding reaction. The rate of disappearance of the species with affinity for binding detergents coincides with the slowest of the two rate-limiting steps detected in the refolding pathway of mAAT. Limited proteolysis studies indicate that the overall structure of the detergent-bound mAAT resembles that of the protein in a complex with GroEL. The mAAT folding intermediates trapped in the presence of detergents can resume reactivation either upon dilution of the detergent below its CMC or by adding beta-cyclodextrin. Thus, isolation of otherwise transient productive folding intermediates for further characterization is possible through the use of detergents. PMID:10082379

  8. Opposite behavior of two isozymes when refolding in the presence of non-ionic detergents.

    PubMed

    Doñate, F; Artigues, A; Iriarte, A; Martinez-Carrion, M

    1998-08-01

    GroEL has a greater affinity for the mitochondrial isozyme (mAAT) of aspartate aminotransferase than for its cytosolic counterpart (cAAT) (Mattingly JR Jr, Iriarte A, Martinez-Carrion M, 1995, J Biol Chem 270:1138-1148), two proteins that share a high degree of sequence similarity and an almost identical spatial structure. The effect of detergents on the refolding of these large, dimeric isozymes parallels this difference in behavior. The presence of non-ionic detergents such as Triton X-100 or lubrol at concentrations above their critical micelle concentration (CMC) interferes with reactivation of mAAT unfolded in guanidinium chloride but increases the yield of cAAT refolding at low temperatures. The inhibitory effect of detergents on the reactivation of mAAT decreases progressively as the addition of detergents is delayed after starting the refolding reaction. The rate of disappearance of the species with affinity for binding detergents coincides with the slowest of the two rate-limiting steps detected in the refolding pathway of mAAT. Limited proteolysis studies indicate that the overall structure of the detergent-bound mAAT resembles that of the protein in a complex with GroEL. The mAAT folding intermediates trapped in the presence of detergents can resume reactivation either upon dilution of the detergent below its CMC or by adding beta-cyclodextrin. Thus, isolation of otherwise transient productive folding intermediates for further characterization is possible through the use of detergents.

  9. Silica Nanoparticles Functionalized with Zwitterionic Sulfobetaine Siloxane for Application as a Versatile Antifouling Coating System.

    PubMed

    Knowles, Brianna R; Wagner, Pawel; Maclaughlin, Shane; Higgins, Michael J; Molino, Paul J

    2017-06-07

    The growing need to develop surfaces able to effectively resist biological fouling has resulted in the widespread investigation of nanomaterials with potential antifouling properties. However, the preparation of effective antifouling coatings is limited by the availability of reactive surface functional groups and our ability to carefully control and organize chemistries at a materials' interface. Here, we present two methods of preparing hydrophilic low-fouling surface coatings through reaction of silica-nanoparticle suspensions and predeposited silica-nanoparticle films with zwitterionic sulfobetaine (SB). Silica-nanoparticle suspensions were functionalized with SB across three pH conditions and deposited as thin films via a simple spin-coating process to generate hydrophilic antifouling coatings. In addition, coatings of predeposited silica nanoparticles were surface functionalized via exposure to zwitterionic solutions. Quartz crystal microgravimetry with dissipation monitoring was employed as a high throughput technique for monitoring and optimizing reaction to the silica-nanoparticle surfaces. Functionalization of nanoparticle films was rapid and could be achieved over a wide pH range and at low zwitterion concentrations. All functionalized particle surfaces presented a high degree of wettability and resulted in large reductions in adsorption of bovine serum albumin protein. Particle coatings also showed a reduction in adhesion of fungal spores (Epicoccum nigrum) and bacteria (Escherichia coli) by up to 87 and 96%, respectively. These results indicate the potential for functionalized nanosilicas to be further developed as versatile fouling-resistant coatings for widespread coating applications.

  10. AN ALTERNATIVE METHOD FOR RELATING MACROSCOPIC TO MICROSCOPIC ACIDITY CONSTANTS WITH ZWITTERIONIC SPECIES

    EPA Science Inventory

    Using the notation of Adams (1916. JACS, 38:1503), zwitterionic microscopic acidity constants defined by: ka = [H+] [+H3NRCOO-]/ [+H3NRCOOH]; kb = [H+] [H2NRCOOH]/ [+H3NRCOOH]; kc = [H+] [H2NRCOO-]/ [+H3NRCOO-]; and kd = [H+] [H2NRCOO-]/ [H2NRCOOH] are historically related to th...

  11. Detergent-associated Solution Conformations of Helical and Beta-barrel Membrane Proteins

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

    Mo, Yiming; Lee, Byung-Kwon; Ankner, John Francis

    2008-01-01

    Membrane proteins present major challenges for structural biology. In particular, the production of suitable crystals for high-resolution structural determination continues to be a significant roadblock for developing an atomic-level understanding of these vital cellular systems. The use of detergents for extracting membrane proteins from the native membrane for either crystallization or reconstitution into model lipid membranes for further study is assumed to leave the protein with the proper fold with a belt of detergent encompassing the membrane-spanning segments of the structure. Small-angle X-ray scattering was used to probe the detergent-associated solution conformations of three membrane proteins, namely bacteriorhodopsin (BR), themore » Ste2p G-protein coupled receptor from Saccharomyces cerevisiae, and the Escherichia coli porin OmpF. The results demonstrate that, contrary to the traditional model of a detergent-associated membrane protein, the helical proteins BR and Ste2p are not in the expected, compact conformation and associated with detergent micelles, while the ?-barrel OmpF is indeed embedded in a disk-like micelle in a properly folded state. The comparison provided by the BR and Ste2p, both members of the 7TM family of helical membrane proteins, further suggests that the interhelical interactions between the transmembrane helices of the two proteins differ, such that BR, like other rhodopsins, can properly refold to crystallize, while Ste2p continues to prove resistant to crystallization from an initially detergent-associated state.« less

  12. Detergent-associated solution conformations of helical and beta-barrel membrane proteins.

    PubMed

    Mo, Yiming; Lee, Byung-Kwon; Ankner, John F; Becker, Jeffrey M; Heller, William T

    2008-10-23

    Membrane proteins present major challenges for structural biology. In particular, the production of suitable crystals for high-resolution structural determination continues to be a significant roadblock for developing an atomic-level understanding of these vital cellular systems. The use of detergents for extracting membrane proteins from the native membrane for either crystallization or reconstitution into model lipid membranes for further study is assumed to leave the protein with the proper fold with a belt of detergent encompassing the membrane-spanning segments of the structure. Small-angle X-ray scattering was used to probe the detergent-associated solution conformations of three membrane proteins, namely bacteriorhodopsin (BR), the Ste2p G-protein coupled receptor from Saccharomyces cerevisiae, and the Escherichia coli porin OmpF. The results demonstrate that, contrary to the traditional model of a detergent-associated membrane protein, the helical proteins BR and Ste2p are not in the expected, compact conformation and associated with detergent micelles, while the beta-barrel OmpF is indeed embedded in a disk-like micelle in a properly folded state. The comparison provided by the BR and Ste2p, both members of the 7TM family of helical membrane proteins, further suggests that the interhelical interactions between the transmembrane helices of the two proteins differ, such that BR, like other rhodopsins, can properly refold to crystallize, while Ste2p continues to prove resistant to crystallization from an initially detergent-associated state.

  13. Neutral vs zwitterionic glycine forms at the water/silica interface: structure, energies, and vibrational features from B3LYP periodic simulations.

    PubMed

    Rimola, Albert; Civalleri, Bartolomeo; Ugliengo, Piero

    2008-12-16

    B3LYP periodic calculations with a triple-xi-polarized Gaussian basis set have been used to study adsorption of glycine on a hydroxylated silica surface (2.2 OH/nm2) model derived from the (001) surface of edingtonite. The simulation envisages glycine adsorbed either as a gas-phase molecule or when microsolvated by up to five H20 molecules. Both neutral and zwitterionic forms of glycine have been considered and their structural, energetic, and spectroscopic vibrational features compared internally and with experiments. As a gas phase glycine sticks in its neutral form at the silica surface, the zwitterion being highly unstable and with transition-state character. When glycine is microsolvated at the silica interface, two H20 molecules render the zwitterion population comparable to that of the neutral form whereas with four H2O molecules the neutral glycine population is wiped out in favor of the zwitterion. With four H20 molecules the most stable structure shows no direct contact between glycine and the silica surface, H20 acting as a mediator via H-bond interactions. The B3LYP energies and structural data were also supported by comparing the scaled harmonic vibrational features with literature FTIR data of glycine adsorbed on an amorphous silica surface either from the gas phase or in water solution.

  14. Refractive index-based determination of detergent concentration and its application to the study of membrane proteins

    PubMed Central

    Strop, Pavel; Brunger, Axel T.

    2005-01-01

    The concentration of detergent in membrane protein preparations can have a critical role on protein stability, function, and the potential for crystallization. Unfortunately, dialysis or protein concentration can lead to an unknown amount of detergent in the final membrane protein preparations. Here we present a method for the determination of detergent concentration based on refractive index of the detergent solution. This method was applied to quantitate the amount of detergent remaining in solution after concentration in various concentrators. We found that the ability of the tested detergents to pass through the molecular weight cutoff membrane correlates well with detergent micelle size. Therefore, the micelle size can be used as a rough guide to estimate the retention of a given detergent in various molecular weight cutoff concentrators. The refractive index method is exceptionally informative when coupled with size exclusion chromatography and light scattering, and can be used to determine the oligomeric state of the membrane protein, the size of a protein-associated micelle, as well as the amount and size of the unbound detergent micelle. PMID:16046633

  15. Refractive index-based determination of detergent concentration and its application to the study of membrane proteins.

    PubMed

    Strop, Pavel; Brunger, Axel T

    2005-08-01

    The concentration of detergent in membrane protein preparations can have a critical role on protein stability, function, and the potential for crystallization. Unfortunately, dialysis or protein concentration can lead to an unknown amount of detergent in the final membrane protein preparations. Here we present a method for the determination of detergent concentration based on refractive index of the detergent solution. This method was applied to quantitate the amount of detergent remaining in solution after concentration in various concentrators. We found that the ability of the tested detergents to pass through the molecular weight cutoff membrane correlates well with detergent micelle size. Therefore, the micelle size can be used as a rough guide to estimate the retention of a given detergent in various molecular weight cutoff concentrators. The refractive index method is exceptionally informative when coupled with size exclusion chromatography and light scattering, and can be used to determine the oligomeric state of the membrane protein, the size of a protein-associated micelle, as well as the amount and size of the unbound detergent micelle.

  16. Development of zwitterionic chromophores for electro-optic applications

    NASA Astrophysics Data System (ADS)

    Xiong, Ying

    In order to unlock the full potential of the zwitterionic NLO chromophores for electro-optic (EO) applications, a new series of PeQDM chromophores with large first hyperpolarizabilities (beta0 ˜ 600 x 10-30 esu) have been designed and synthesized. A large EO coefficient (r33) of 110 pm/V at 1550 nm has been realized with a 5 wt% (corresponding to 3.8 wt% core content) chromophore doped polymer. The EO study of guest-host polymers reveals that dipolar dye aggregation in a less polar medium is responsible for a low chromophore loading and low EO activity. Modification of NLO chromophore by attaching large dendrons can effectively increase the chromophore loading in a host and improve the poling efficiency. Crosslinkable NLO polymers have also been prepared to improve the temporal stability of the poled noncentrosymmetric order. The following are some important highlights from this thesis work. (1) A series of thermally stable zwitterionic chromophores (PeQDM) with large first hyperpolarizabilities (beta up to -1797 x 10-30 esu) are synthesized in good yields (˜ 50%). The charge-separated ground state is evident by a negative solvatochromism. X-ray crystallographic data further confirms the zwitterionic nature and demonstrates a face-to-face anti-parallel H-aggregation of two monomers due to strong electrostatic interactions between the dipoles. (2) PeQDM chromophores are also NIR fluorescent (lambdaPL ˜ 840-870 nm in solution) and labile to acid, making them potential candidates for NIR pH sensor applications. (3) The hydroxyl-containing PeQDM chromophores are modified with ES-dendron, which exhibit good solubility in solvents and polymers. Self-forming films can be prepared by direct casting or spin-coating of two dendrons modified chromophores (ES-PeQDM-2 and ES-PeQDM-3), in which the chromophore core contents reach 14.9 and 16.9 wt%, respectively. Compared to ES-PeQDM-2 with two dendrons only at the donor part (r33 = 0 pm/V), ES-PeQDM-3 with the bulky ES

  17. Deuterated detergents for structural and functional studies of membrane proteins: Properties, chemical synthesis and applications.

    PubMed

    Hiruma-Shimizu, Kazumi; Shimizu, Hiroki; Thompson, Gary S; Kalverda, Arnout P; Patching, Simon G

    2015-01-01

    Detergents are amphiphilic compounds that have crucial roles in the extraction, purification and stabilization of integral membrane proteins and in experimental studies of their structure and function. One technique that is highly dependent on detergents for solubilization of membrane proteins is solution-state NMR spectroscopy, where detergent micelles often serve as the best membrane mimetic for achieving particle sizes that tumble fast enough to produce high-resolution and high-sensitivity spectra, although not necessarily the best mimetic for a biomembrane. For achieving the best quality NMR spectra, detergents with partial or complete deuteration can be used, which eliminate interfering proton signals coming from the detergent itself and also eliminate potential proton relaxation pathways and strong dipole-dipole interactions that contribute line broadening effects. Deuterated detergents have also been used to solubilize membrane proteins for other experimental techniques including small angle neutron scattering and single-crystal neutron diffraction and for studying membrane proteins immobilized on gold electrodes. This is a review of the properties, chemical synthesis and applications of detergents that are currently commercially available and/or that have been synthesized with partial or complete deuteration. Specifically, the detergents are sodium dodecyl sulphate (SDS), lauryldimethylamine-oxide (LDAO), n-octyl-β-D-glucoside (β-OG), n-dodecyl-β-D-maltoside (DDM) and fos-cholines including dodecylphosphocholine (DPC). The review also considers effects of deuteration, detergent screening and guidelines for detergent selection. Although deuterated detergents are relatively expensive and not always commercially available due to challenges associated with their chemical synthesis, they will continue to play important roles in structural and functional studies of membrane proteins, especially using solution-state NMR.

  18. 40 CFR 417.150 - Applicability; description of the manufacture of spray dried detergents subcategory.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... manufacture of spray dried detergents subcategory. 417.150 Section 417.150 Protection of Environment... POINT SOURCE CATEGORY Manufacture of Spray Dried Detergents Subcategory § 417.150 Applicability; description of the manufacture of spray dried detergents subcategory. The provisions of this subpart are...

  19. 40 CFR 417.180 - Applicability; description of the manufacture of drum dried detergents subcategory.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... manufacture of drum dried detergents subcategory. 417.180 Section 417.180 Protection of Environment... POINT SOURCE CATEGORY Manufacture of Drum Dried Detergents Subcategory § 417.180 Applicability; description of the manufacture of drum dried detergents subcategory. The provisions of this subpart are...

  20. Combining Fullerenes and Zwitterions in non-Conjugated Polymer Interlayers to Raise Solar Cell Efficiency.

    PubMed

    Liu, Yao; Sheri, Madhu; Cole, Marcus D; Emrick, Todd; Russell, Thomas P

    2018-06-12

    Polymer zwitterions were synthesized by nucleophilic ring-opening of 3,3'-(but-2-ene-1,4-diyl)bis(1,2-oxathiolane 2,2-dioxide) (a bis-sultone) with functional perylene diimide (PDI) or fullerene monomers. Integration of these polymers into solar cell devices as cathode interlayers boosted efficiencies of fullerene-based organic photovoltaics (OPVs) from 2.75% to 10.74%, and of non-fullerene-based OPVs from 4.25% to 10.10%, demonstrating the versatility of these interlayer materials in OPVs. The fullerene-containing polymer zwitterion (C60-PZ) showed a higher interfacial dipole (∆) value and electron mobility than its PDI counterpart (PDI-PZ), affording solar cells with high efficiency. The power of PDI-PZ and C60-PZ to improve electron injection and extraction processes when positioned between metal electrodes and organic semiconductors highlights their promise to overcome energy barriers at the hard-soft materials interface of organic electronics. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Comparative lipidomics and proteomics analysis of platelet lipid rafts using different detergents.

    PubMed

    Rabani, Vahideh; Davani, Siamak; Gambert-Nicot, Ségolène; Meneveau, Nicolas; Montange, Damien

    2016-11-01

    Lipid rafts play a pivotal role in physiological functions of platelets. Their isolation using nonionic mild detergents is considered as the gold standard method, but there is no consensual detergent for lipid raft studies. We aimed to investigate which detergent is the most suitable for lipid raft isolation from platelet membrane, based on lipidomics and proteomics analysis. Platelets were obtained from healthy donors. Twelve sucrose fractions were extracted by three different detergents, namely Brij 35, Lubrol WX, and Triton X100, at 0.05% and 1%. After lipidomics analysis and determination of fractions enriched in cholesterol (Ch) and sphingomyelin (SM), proteomics analysis was performed. Lipid rafts were mainly observed in 1-4 fractions, and non-rafts were distributed on 5-12 fractions. Considering the concentration of Ch and SM, Lubrol WX 1% and Triton X100 1% were more suitable detergents as they were able to isolate lipid raft fractions that were more enriched than non-raft fractions. By proteomics analysis, overall, 822 proteins were identified in platelet membrane. Lipid raft fractions isolated with Lubrol WX 0.05% and Triton X100 1% contained mainly plasma membrane proteins. However, only Lubrol WX 0.05 and 1% and Triton X100 1% were able to extract non-denaturing proteins with more than 10 transmembrane domains. Our results suggest that Triton X100 1% is the most suitable detergent for global lipid and protein studies on platelet plasma membrane. However, the detergent should be adapted if investigation of an association between specific proteins and lipid rafts is planned.

  2. Characterization of Protein Detergent Complexes by NMR, Light Scattering, and Analytical Ultracentrifugation

    PubMed Central

    Maslennikov, Innokentiy; Krupa, Martin; Dickson, Christopher; Esquivies, Luis; Blain, Katherine; Kefala, Georgia; Choe, Senyon; Kwiatkowski, Witek

    2009-01-01

    Bottlenecks in expression, solubilization, purification and crystallization hamper the structural study of integral membrane proteins (IMPs). Successful crystallization is critically dependent on the purity, stability and oligomeric homogeneity of an IMP sample. These characteristics are in turn strongly influenced by the type and concentration of the detergents used in IMP preparation. By utilizing the techniques and analytical tools we earlier developed for the characterization of protein-detergent complexes (PDCs) (Maslennikov et al., 2007), we demonstrate that for successful protein extraction from E. coli membrane fractions, the solubilizing detergent associates preferentially to IMPs rather than to membrane lipids. Notably, this result is contrary to the generally accepted mechanism of detergent-mediated IMP solubilization. We find that for one particular member of the family of proteins studied (E. coli receptor kinases, which is purified in mixed multimeric states and oligomerizes through its transmembrane region), the protein oligomeric composition is largely unaffected by a 10-fold increase in protein concentration, by alteration of micelle properties through addition of other detergents to the PDC sample, or by a 20-fold variation in the detergent concentration used for solubilization of the IMP from the membrane. We observed that the conditions used for expression of the IMP, which impact protein density in the membrane, has the greatest influence on the IMP oligomeric structure. Finally, we argue that for concentrating PDCs smaller than 30 kDa, stirred concentration cells are less prone to over-concentration of detergent and are therefore more effective than centrifugal ultrafiltration devices. PMID:19214777

  3. Microwave-assisted micellar extraction of organic and inorganic iodines using zwitterionic surfactants.

    PubMed

    Wang, Shu-Ling; Yi, Ling; Ye, Li-Hong; Cao, Jun; Du, Li-Jing; Peng, Li-Qing; Xu, Jing-Jing; Zhang, Qi-Dong

    2017-08-04

    Zwitterionic surfactant, used as extractant in microwave-assisted extraction (MAE) was investigated for the first time to extract organic and inorganic iodines from kelp samples. Optimized conditions for the MAE were 200W of microwave irradiation power, 100°C of extraction temperature, 10min of microwave irradiation time, 1g of sample, and 20mL of solvent volume. Ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF/MS) was used for the quantitative and qualitative analyses of the iodines. Under the optimum experimental conditions, KI, MIT and DIT were identified in kelp samples, the limits of detection of these analytes were ranged between 3.39 and 6.31ng/mL. The recoveries for spiked samples obtained from different areas were all higher than 92.48%. Compared with the ultrasound-assisted extraction, the proposed method is faster and more effective. Thus, the combination of zwitterionic surfactant-MAE and UHPLC-Q-TOF/MS made up a simple, rapid and effective approach for extraction and determination of iodine compounds in complex seaweed materials. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Detergent-dependent kinetics of truncated Plasmodium falciparum dihydroorotate dehydrogenase.

    PubMed

    Malmquist, Nicholas A; Baldwin, Jeffrey; Phillips, Margaret A

    2007-04-27

    The survival of the malaria parasite Plasmodium falciparum is dependent upon the de novo biosynthesis of pyrimidines. P. falciparum dihydroorotate dehydrogenase (PfDHODH) catalyzes the fourth step in this pathway in an FMN-dependent reaction. The full-length enzyme is associated with the inner mitochondrial membrane, where ubiquinone (CoQ) serves as the terminal electron acceptor. The lipophilic nature of the co-substrate suggests that electron transfer to CoQ occurs at the two-dimensional lipid-solution interface. Here we show that PfDHODH associates with liposomes even in the absence of the N-terminal transmembrane-spanning domain. The association of a series of ubiquinone substrates with detergent micelles was studied by isothermal titration calorimetry, and the data reveal that CoQ analogs with long decyl (CoQ(D)) or geranyl (CoQ(2)) tails partition into detergent micelles, whereas that with a short prenyl tail (CoQ(1)) remains in solution. PfDHODH-catalyzed reduction of CoQ(D) and CoQ(2), but not CoQ(1), is stimulated as detergent concentrations (Tween 80 or Triton X-100) are increased up to their critical micelle concentrations, beyond which activity declines. Steady-state kinetic data acquired for the reaction with CoQ(D) and CoQ(2) in substrate-detergent mixed micelles fit well to a surface dilution kinetic model. In contrast, the data for CoQ(1) as a substrate were well described by solution steady-state kinetics. Our results suggest that the partitioning of lipophilic ubiquinone analogues into detergent micelles needs to be an important consideration in the kinetic analysis of enzymes that utilize these substrates.

  5. Respiratory symptoms and pulmonary function impairment among detergent plant workers in Jos, Northern Nigeria.

    PubMed

    Babashani, M; Iliyasu, Z; Ukoli, C O

    2008-01-01

    The industrial process of detergent production could be deleterious to lung function. This study describes respiratory symptoms and ventilatory function impairment among detergent workers in Jos, Northern Nigeria. Two hundred detergent plant workers and controls were studied for the presence of respiratory symptoms and ventilatory function impairment using the MRC questionnaire and Spirometry. A significantly higher proportion of exposed detergent workers 178 (87.0%) reported respiratory symptoms compared to 52 (26.0%) controls [OR=23; 95% CI=12.9-41.3] (P<0.001). Commonest symptoms include rhinitis (57.5% versus 11.0%) and cough (48.5% versus 15%). Symptoms were most prevalent in the packaging section. FEV1, FVC and PEFR were significantly reduced among exposed detergent workers. Similarly, the predicted values of PEFR, FVC and FEV1, were significantly reduced among smokers (P<0. 001). Respiratory symptoms are highly prevalent among detergent workers. This was associated with impaired pulmonary function. Protective equipment and periodic lung function tests could reduce these effects.

  6. Effects of household detergent on anaerobic fermentation of kitchen wastewater from food waste disposer.

    PubMed

    Lee, K H; Park, K Y; Khanal, S K; Lee, J W

    2013-01-15

    This study examines the effects of household detergent on anaerobic methane fermentation of wastewater from food waste disposers (FWDs). Anaerobic toxicity assay (ATA) demonstrated that methane production substantially decreased at a higher detergent concentration. The Gompertz three-parameter model fitted well with the ATA results, and both the extent of methane production (M) and methane production rate (R(m)) obtained from the model were strongly affected by the concentration of the detergent. The 50% inhibitory concentration (IC(50)) of the detergent was 603 mg/L based on R(m). Results from fatty acid methyl esters (FAMEs) analysis of microbial culture revealed that deterioration of methane fermentation was attributed to impaired structure of anaerobic microbial membrane due to detergent. This study suggests that wastewater from FWD could be used for methane production, but it is necessary to reduce the concentration of detergent prior to anaerobic fermentation. Copyright © 2012 Elsevier B.V. All rights reserved.

  7. Evaluation of Liquid Detergents and Methods Used for Airfield Rubber Removal

    DTIC Science & Technology

    2012-07-31

    friction. Each detergent is applied to the pavement surface in controlled 50’ x 30’ patches. There were three patches placed for each detergent with...Measurements ..........................................................................................................16 6.3. Periodic Pavement Wetting...19 Figure 14. Pre-Cleaning Micro -Texture Data Graph (GT View Data

  8. Amino acid-based zwitterionic polymers: antifouling properties and low cytotoxicity.

    PubMed

    Li, Wenchen; Liu, Qingsheng; Liu, Lingyun

    2014-01-01

    A group of five amino acid containing zwitterionic vinyl monomers, based on serine, lysine, ornithine, glutamic acid, and aspartic acid, respectively, were proposed and developed for potential antifouling applications. Their polymer brushes were grafted on gold chips by surface-initiated photoiniferter-mediated polymerization. We then compared their performance in resisting protein adsorption from full human serum and plasma. All five polymers can reduce protein adsorption by more than 90% compared to the unmodified gold. The ornithine-based and aspartic acid-based poly(methacrylamide) can most strongly resist protein adsorption from serum and plasma, compared to the other three. The ability of surfaces to suppress bacterial adhesion is another criterion in evaluating antifouling properties of materials. Our results show that the five polymer-grafted surfaces can significantly suppress Escherichia coli K12 adhesion to 99% compared to the bare gold surface. The zwitterionic structure of amino acids, with homogenously distributed and balanced positive and negative charges, is responsible for the outstanding antifouling properties. Considering multiple potential applications (e.g. medical devices and drug delivery) of the antifouling materials, we further systematically evaluated the cytotoxicity of both monomers and polymer nanogels for all five materials at various concentrations. Very low cytotoxicity was observed for all tested amino acid-based monomers and nanogels, which is comparable or even lower than the traditional and some newly developed antifouling materials, which might be related to the biomimetic nature of amino acids.

  9. Detection of antibiotics in goat's milk: effect of detergents on the response of microbial inhibitor tests.

    PubMed

    Romero, Tamara; Beltrán, María Carmen; Althaus, Rafael Lisandro; Molina, María Pilar

    2014-08-01

    The aim of the study was to evaluate the interference of acid and alkaline detergents employed in the cleaning of milking equipment of caprine dairy farms on the performance of microbial tests used in antibiotic control (BRT MRL, Delvotest MCS, and Eclipse 100). Eight concentrations of commercial detergents, five acid (0-0.25%) and five alkaline (0-1%) were add to antimicrobial-free goat's milk to evaluate the detergent effect on the response of microbial inhibitor tests. To evaluate the effect of detergents on the detection capability of microbial tests two detergents at 0.5 ml/l (one acid and one basic) and eight concentrations of four β-lactam antibiotics (ampicillin, amoxicillin, cloxacillin and benzylpenicillin) were used. Milk without detergents was used as control. The spiked samples were analysed twelve times by three microbial tests. The results showed that the presence of acid detergents did not affect the response of microbial tests for any of the concentrations tested. However, at concentrations equal to or greater than 2 ml/l alkaline detergents positive results were found in microbial tests (16.7-100%). The detection limits of the screening tests for penicillins were not modified substantially by the presence of detergents. In general, the presence of acid and alkaline detergents in goat's milk did not produce a great interference in the microbial tests, only high concentrations of detergents could cause non-compliant results, but these concentrations are difficult to find in practice if proper cleaning procedures are applied in goat dairy farms.

  10. Ingestion of Laundry Detergent Packets in Children.

    PubMed

    Shah, Lindsey Wilson

    2016-08-01

    Ingestion of laundry detergent packets is an important threat to young children. Because of their developmental stage, toddlers are prone to place these small, colorful packets in their mouths. The packets can easily burst, sending a large volume of viscous, alkaline liquid throughout the oropharynx. Ingestion causes major toxic effects, including depression of the central nervous system, metabolic acidosis, respiratory distress, and dysphagia. Critical care nurses should anticipate these clinical effects and facilitate prompt intervention. Increased understanding of the risks and clinical effects of ingestion of laundry detergent packets will better prepare critical care nurses to provide care for these children. (Critical Care Nurse 2016; 36[4]:70-75). ©2016 American Association of Critical-Care Nurses.

  11. The safety of synthetic zeolites used in detergents.

    PubMed

    Fruijtier-Pölloth, Claudia

    2009-01-01

    Synthetic zeolites are replacing phosphates as builders in laundry detergents; workers and consumers may, therefore, increasingly be exposed to these materials and it is important to assess their safety. This article puts mechanistic, toxicological and exposure data into context for a safety assessment. Zeolites are hygroscopic compounds with ion-exchanging properties. They may partially decompose under acidic conditions such as in the stomach releasing sodium ions, silicic acid and aluminum salts. The intact molecule is not bioavailable after oral intake or exposure through the dermal and inhalational routes. Under current conditions of manufacture and use, no systemic toxicity is to be expected from neither the intact molecule nor the degradation products; a significant effect on the bioavailability of other compounds is not likely. Zeolites may cause local irritation. It is, therefore, important to minimise occupational exposure. The co-operation of detergent manufacturers with the manufacturers of washing machines is necessary to find the right balance between environmental aspects such as energy and water savings and the occurrence of detergent residues on textiles due to insufficient rinsing.

  12. Effects of a detergent micelle environment on P-glycoprotein (ABCB1)-ligand interactions

    PubMed Central

    Shukla, Suneet; Abel, Biebele; Chufan, Eduardo E.; Ambudkar, Suresh V.

    2017-01-01

    P-glycoprotein (P-gp) is a multidrug transporter that uses energy from ATP hydrolysis to export many structurally dissimilar hydrophobic and amphipathic compounds, including anticancer drugs from cells. Several structural studies on purified P-gp have been reported, but only limited and sometimes conflicting information is available on ligand interactions with the isolated transporter in a dodecyl-maltoside detergent environment. In this report we compared the biochemical properties of P-gp in native membranes, detergent micelles, and when reconstituted in artificial membranes. We found that the modulators zosuquidar, tariquidar, and elacridar stimulated the ATPase activity of purified human or mouse P-gp in a detergent micelle environment. In contrast, these drugs inhibited ATPase activity in native membranes or in proteoliposomes, with IC50 values in the 10–40 nm range. Similarly, a 30–150-fold decrease in the apparent affinity for verapamil and cyclic peptide inhibitor QZ59-SSS was observed in detergent micelles compared with native or artificial membranes. Together, these findings demonstrate that the high-affinity site is inaccessible because of either a conformational change or binding of detergent at the binding site in a detergent micelle environment. The ligands bind to a low-affinity site, resulting in altered modulation of P-gp ATPase activity. We, therefore, recommend studying structural and functional aspects of ligand interactions with purified P-gp and other ATP-binding cassette transporters that transport amphipathic or hydrophobic substrates in a detergent-free native or artificial membrane environment. PMID:28283574

  13. Manifestation of cryptic fibroblast tissue factor occurs at detergent concentrations which dissolve the plasma membrane.

    PubMed

    Carson, S D

    1996-04-01

    Cultured fibroblasts treated with increasing concentrations of detergents expressed only encrypted levels of tissue factor activity (measured by fX activation in the presence of fVIIa), characteristic of undamaged cells, until each detergent reached a critical concentration at which the cryptic tissue factor activity was manifested. Beyond the narrow ranges of concentrations over which the detergents stimulated tissue factor activity, the detergents were inhibitory. Studies with Triton X-100 and octyl glucoside revealed that manifestation of tissue factor activity coincided with breakdown of the plasma membrane. The magnitude of the increased tissue factor activity differed among detergents, with octyl glucoside giving the largest response. The tissue factor that was active after Triton X-100 treatment remained mostly associated with the insoluble cell residue, whereas the concentration of octyl glucoside which stimulated activity released tissue factor activity into the supernatant. Radiolabeled antibody against human tissue factor was used to show that a small percentage of the total accessible tissue factor remained in the insoluble fraction after treatment with either non-ionic detergent. Chromatographic analysis of lipids extracted from cells treated with detergents and dansyl chloride showed dansyl-reactivity of phosphatidylserine on intact cells, and solubilization of membrane lipids at sublytic concentrations of detergents. These findings reveal that there is a critical level of detergent-induced membrane damage at which tissue factor activity is maximally expressed, in essentially an all-or-none manner. The results are consistent with a major role for phospholipid asymmetry in regulation of tissue factor specific activity, but require either maintenance of asymmetry during sublytic detergent perturbation of the plasma membrane or additional control mechanisms.

  14. Influence of detergents on water drift in cooling towers

    NASA Astrophysics Data System (ADS)

    Vitkovicova, Rut

    An influence of detergents on the water drift from the cooling tower was experimentally investigated. For this experimental measurements was used a model cooling tower, especially an experimental aerodynamic line, which is specially designed for the measurement and monitoring of processes taking place around the eliminators of the liquid phase. The effect of different concentrations of detergent in the cooling water on the drift of water droplets from a commonly used type eliminator was observed with visualization methods.

  15. Fluorinated diglucose detergents for membrane-protein extraction.

    PubMed

    Boussambe, Gildas Nyame Mendendy; Guillet, Pierre; Mahler, Florian; Marconnet, Anaïs; Vargas, Carolyn; Cornut, Damien; Soulié, Marine; Ebel, Christine; Le Roy, Aline; Jawhari, Anass; Bonneté, Françoise; Keller, Sandro; Durand, Grégory

    2018-05-29

    Fluorinated surfactants have scarcely been explored for the direct extraction of proteins from membranes because fluorination is believed to abrogate detergency. However, we have recently shown that a commercially available fluorinated surfactant readily solubilizes lipid membranes, thereby suggesting that fluorination per se does not interfere with detergent activity. In this work, we developed new fluorinated surfactants that exhibit detergency in terms of both lipid-vesicle solubilization and membrane-protein extraction. The compounds made and tested contain two glucose moieties as polar headgroup, a hydrogenated thioether linker, and a perfluorinated alkyl tail with either 4, 6, or 8 carbon atoms. The physicochemical properties of the micelles formed by the three fluorinated surfactants were evaluated by NMR spectroscopy, surface tensiometry, isothermal titration calorimetry, dynamic light scattering, small-angle X-ray scattering, and analytical ultracentrifugation. At 25°C, micellization was mainly entropy-driven, and the CMC values were found to decrease with chain length of the fluorinated tail, whereas the aggregation number increased with chain length. Remarkably, all three surfactants were found to solubilize lipid vesicles and extract a broad range of proteins from Escherichiacoli membranes. These findings demonstrate, for the first time, that nonionic fluorinated surfactants could be further exploited for the direct extraction and solubilization of membrane proteins. Copyright © 2018. Published by Elsevier Inc.

  16. Laundry pod and non-pod detergent related emergency department visits occurring in children in the USA.

    PubMed

    Swain, Thomas A; McGwin, Gerald; Griffin, Russell

    2016-12-01

    Previous studies have reported that children are at risk of severe injuries from exposure to laundry detergent pods. For the first time, this study sought to compare demographic and exposure characteristics and risk among children exposed to pod and non-pod laundry detergents presenting to emergency departments (EDs). Data from the National Electronic Injury Surveillance System (NEISS) from 2012-2014 were used. All observations with injuries involving laundry detergent (NEISS code 0949) were included in this study. The χ 2 test was used for bivariate analysis and logistic regression was used to determine the OR and 95% CI of hospitalisation for pod related versus non-pod laundry detergent exposures. From 2012-2014, there were an estimated 26 062 non-pod and 9814 pod laundry detergent related exposures among those aged 18 years and younger. For pod detergent, children aged 0-5 years had the most injuries. Poisoning (71.3%) was the most common diagnosis for pod detergent while contact dermatitis (72.2%) was most common for non-pod detergent. Hospitalisation occurred in 12.5% of pod detergent cases and just 3.0% of non-pod cases. Compared with non-pod detergent, those exposed to pod detergent were 4 times as likely to be hospitalised (OR 4.02; 95% CI 1.96 to 8.24). A greater effort should be made to appropriately educate the public about the dangers of laundry detergents, specifically pods, so a safe home environment can be established. While new regulations such as childproof containers, opaque packaging, and less appealing and colourful pods could reduce the number of pod related ED visits for children, caregivers should store detergents, along with other chemicals, in a secure location where children cannot easily access them. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.

  17. Hemocompatible control of sulfobetaine-grafted polypropylene fibrous membranes in human whole blood via plasma-induced surface zwitterionization.

    PubMed

    Chen, Sheng-Han; Chang, Yung; Lee, Kueir-Rarn; Wei, Ta-Chin; Higuchi, Akon; Ho, Feng-Ming; Tsou, Chia-Chun; Ho, Hsin-Tsung; Lai, Juin-Yih

    2012-12-21

    In this work, the hemocompatibility of zwitterionic polypropylene (PP) fibrous membranes with varying grafting coverage of poly(sulfobetaine methacrylate) (PSBMA) via plasma-induced surface polymerization was studied. Charge neutrality of PSBMA-grafted layers on PP membrane surfaces was controlled by the low-pressure and atmospheric plasma treatment in this study. The effects of grafting composition, surface hydrophilicity, and hydration capability on blood compatibility of the membranes were determined. Protein adsorption onto the different PSBMA-grafted PP membranes from human fibrinogen solutions was measured by enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies. Blood platelet adhesion and plasma clotting time measurements from a recalcified platelet-rich plasma solution were used to determine if platelet activation depends on the charge bias of the grafted PSBMA layer. The charge bias of PSBMA layer deviated from the electrical balance of positively and negatively charged moieties can be well-controlled via atmospheric plasma-induced interfacial zwitterionization and was further tested with human whole blood. The optimized PSBMA surface graft layer in overall charge neutrality has a high hydration capability and keeps its original blood-inert property of antifouling, anticoagulant, and antithrmbogenic activities when it comes into contact with human blood. This work suggests that the hemocompatible nature of grafted PSBMA polymers by controlling grafting quality via atmospheric plasma treatment gives a great potential in the surface zwitterionization of hydrophobic membranes for use in human whole blood.

  18. Effect of soaps and detergents on epidermal barrier function.

    PubMed

    Wolf, Ronni; Parish, Lawrence Charles

    2012-01-01

    The past decade has witnessed an explosion of new impartial information about the complex interaction of the skin with topically applied substances, including soaps and detergents. Despite of all these new data, our knowledge on the exact pathomechanism and molecular events leading to detergent-induced barrier dysfunction remains incomplete and the answers continue to elude us. The longtime prevailing opinion which contends that the damaging effect of soaps and detergents is related to their property to extract and remove useful intercellular lipids has mostly been abandoned. Although this effect might be involved in the damaging effect, it is definitely not the sole mechanism, nor, indeed, is it even the main one. Skin proteins damage, the interaction with keratins and their denaturation, swelling of cell membranes and collagen fibers, cytotoxicity expressed with cellular lysis are other important mechanisms. One proposed mechanism is that an initial stratum corneum hyper-hydration results from a continuous disruption of the secondary and tertiary structures of keratin protein by surfactants, exposing new water-binding sites, thereby increasing the hydration of the membrane. Following evaporation of excess water, the denatured keratin possesses a decreased water-binding capacity and decreased ability to function as a barrier. Recent studies have also emphasized the effects of detergents on lipid synthesis, on lipid-metabolizing enzymes and on keratinocyte differentiation. Copyright © 2012 Elsevier Inc. All rights reserved.

  19. Effects of detergents on ribosomal precursor subunits of Bacillus megaterium.

    PubMed

    Body, A; Brownstein, B H

    1978-01-01

    Cell extracts prepared by osmotic lysis of protoplasts were analyzed by sucrose gradient sedimentation. In the absence of detergents, ribosomal precursor particles were found in a gradient fraction which sedimented faster than mature 50S subunits and in two other fractions coincident with mature 50S and 30S ribosomal subunits. Phospholipid, an indicator of membrane, was shown to be associated with only the fastest-sedimenting ribosomal precursor particle fraction. After the extracts were treated with detergents, all phospholipid was found at the top of the gradients. Brij 58, Triton X-100, and Nonidet P-40 did not cause a change in the sedimentation values of precursors; however, the detergents deoxycholate or LOC (Amway Corp.) disrupted the fastest-sedimenting precursor and converted the ribosomal precursor subunits which sedimented at the 50S and 30S positions to five different classes of more slowly sedimenting particles. Earlier reports on the in vivo assembly of ribosomal subunits have shown that several stages of ribosomal precursor subunits exist, and, in the presence of the detergents deoxycholate and LOC, which had been used to prepare cell extracts, the precursors sedimented more slowly. Our data are consistent with the hypothesis that those detergents selectively modify the structure of ribosomal precursors and lend further support to the hypothesis that the in vivo ribosomal precursor subunits have 50S and 30S sedimentation values. In addition, these data support the idea that the ribosomal precursor particles found in the fast-sedimenting fraction may constitute a unique precursor fraction.

  20. Effects of Detergents on Ribosomal Precursor Subunits of Bacillus megaterium

    PubMed Central

    Body, Barbara A.; Brownstein, Bernard H.

    1978-01-01

    Cell extracts prepared by osmotic lysis of protoplasts were analyzed by sucrose gradient sedimentation. In the absence of detergents, ribosomal precursor particles were found in a gradient fraction which sedimented faster than mature 50S subunits and in two other fractions coincident with mature 50S and 30S ribosomal subunits. Phospholipid, an indicator of membrane, was shown to be associated with only the fastest-sedimenting ribosomal precursor particle fraction. After the extracts were treated with detergents, all phospholipid was found at the top of the gradients. Brij 58, Triton X-100, and Nonidet P-40 did not cause a change in the sedimentation values of precursors; however, the detergents deoxycholate or LOC (Amway Corp.) disrupted the fastest-sedimenting precursor and converted the ribosomal precursor subunits which sedimented at the 50S and 30S positions to five different classes of more slowly sedimenting particles. Earlier reports on the in vivo assembly of ribosomal subunits have shown that several stages of ribosomal precursor subunits exist, and, in the presence of the detergents deoxycholate and LOC, which had been used to prepare cell extracts, the precursors sedimented more slowly. Our data are consistent with the hypothesis that those detergents selectively modify the structure of ribosomal precursors and lend further support to the hypothesis that the in vivo ribosomal precursor subunits have 50S and 30S sedimentation values. In addition, these data support the idea that the ribosomal precursor particles found in the fast-sedimenting fraction may constitute a unique precursor fraction. PMID:412833

  1. Membrane proteins, detergents and crystals: what is the state of the art?

    PubMed Central

    Loll, Patrick J.

    2014-01-01

    At the time when the first membrane-protein crystal structure was determined, crystallization of these molecules was widely perceived as extremely arduous. Today, that perception has changed drastically, and the process is regarded as routine (or nearly so). On the occasion of the International Year of Crystallography 2014, this review presents a snapshot of the current state of the art, with an emphasis on the role of detergents in this process. A survey of membrane-protein crystal structures published since 2012 reveals that the direct crystallization of protein–detergent complexes remains the dominant method­ology; in addition, lipidic mesophases have proven immensely useful, particularly in specific niches, and bicelles, while perhaps undervalued, have provided important contributions as well. Evolving trends include the addition of lipids to protein–detergent complexes and the gradual incorporation of new detergents into the standard repertoire. Stability has emerged as a critical parameter controlling how a membrane protein behaves in the presence of detergent, and efforts to enhance stability are discussed. Finally, although discovery-based screening approaches continue to dwarf mechanistic efforts to unravel crystallization, recent technical advances offer hope that future experiments might incorporate the rational manipulation of crystallization behaviors. PMID:25484203

  2. 40 CFR 80.156 - Liability for violations of the interim detergent program controls and prohibitions.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ..., or detergent-additized post-refinery component satisfied relevant requirements when it left their... detergent satisfied all relevant requirements when it left the detergent manufacturer's control; and (B... manufactured. (2) The test results must accurately establish that, when it left the manufacturer's control, the...

  3. Interactions of Polyethylenimines with Zwitterionic and Anionic Lipid Membranes.

    PubMed

    Kwolek, Urszula; Jamróz, Dorota; Janiczek, Małgorzata; Nowakowska, Maria; Wydro, Paweł; Kepczynski, Mariusz

    2016-05-17

    Interactions between polyethylenimines (PEIs) and phospholipid membranes are of fundamental importance for various biophysical applications of these polymers such as gene delivery. Despite investigations into the nature of these interactions, their molecular basis remains poorly understood. In this article, we combined experimental methods and atomistic molecular dynamics (MD) simulations to obtain comprehensive insight into the effect of linear and branched PEIs on zwitterionic and anionic bilayers used as simple models of mammalian cellular membranes. Our results show that PEIs adsorb only partially on the surface of zwitterionic membranes by forming hydrogen bonds to the lipid headgroups, whereas a large part of the polymer chains dangles freely in the aqueous phase. In contrast, PEIs readily adhere to and insert into the anionic membrane. The attraction of the polymer chains to the membrane is due to electrostatic interactions as well as hydrogen bonding between the amine groups of PEI and the phosphate groups of lipids. These interactions were found to induce a substantial reorganization of the bilayer in the polymer vicinity due to the reorientation of lipid molecules. The lipid headgroups were pulled toward the center of the membrane, which can facilitate transmembrane translocations of anionic lipids. Furthermore, the PEI-lipid interactions affect the stability of liposomal dispersions, but we did not see any evidence of disruption of the vesicular structures into small fragments at polymer concentrations typically used in gene therapy. Our results provide a detailed molecular-level description of the lipid organization in the membrane in the presence of polycations that can be useful in understanding their mechanisms of in vitro and in vivo cytotoxicity.

  4. Cloud Condensation Nuclei in Cumulus Humilis - Selected Case Study During the CHAPS Campaign

    NASA Astrophysics Data System (ADS)

    Yu, X.; Berg, L. K.; Berkowitz, C. M.; Alexander, M. L.; Lee, Y.; Laskin, A.; Ogren, J. A.; Andrews, B.

    2009-12-01

    The Cumulus Humilis Aerosol Processing Study (CHAPS) provided a unique opportunity to study aerosol and cloud processing. Clouds play an active role in the processing and cycling of atmospheric constituents. Gases and particles can partition to cloud droplets by absorption and condensation as well as activation and pact scavenging. The Department of Energy (DOE) G-1 aircraft was used as one of the main platforms in CHAPS. Flight tracks were designed and implemented to characterize freshly emitted aerosols on cloud top and cloud base as well as with cloud, i.e., cumulus humilis (or fair-weather cumulus), in the vicinity of Oklahoma City. Measurements of interstitial aerosols and residuals of activated condensation cloud nuclei were conducted simultaneously. The interstitial aerosols were determined downstream of an isokinetic inlet; and the activated particles downstream of a counter-flow virtual impactor (CVI). The sampling line to the Aerodyne Aerosol Mass Spectrometer was switched between the isokinetic inlet and the CVI to allow characterization of interstitial particles out of clouds in contrast to particles activated in clouds. Trace gases including ozone, carbon monoxide, sulfur dioxide, and a series of volatile organic compounds (VOCs) were also measured as were key meteorological state parameters including liquid water content, cloud drop size, and dew point temperature were measured. This work will focus on studying CCN properties in cumulus humilis. Several approaches will be taken. The first is single particle analysis of particles collected by the Time-Resolved Aerosol Sampler (TRAC) by SEM/TEM coupled with EDX. We will specifically look into differences in particle properties such as chemical composition and morphology between activated and interstitial ones. The second analysis will link in situ measurements with the snap shots observations by TRAC. For instance, by looking into the characteristic m/z obtained by AMS vs. CO or isoprene, one can gain

  5. Photodetachment of Zwitterions: Probing Intramolecular Coulomb Repulsion and Attraction in the Gas Phase Using Pyridinium Dicarboxylate Anions

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

    Wang, Xue B.; Dacres, J E.; Yang, Xin

    2003-10-23

    Zwitterions are critically important in many biological transformations and are used in numerous chemical processes. The consequences of electrostatic effects on reactivity and physical properties, however, are largely unknown. In this work, we report the results of negative ion photoelectron spectra of nine isomeric pyridinium dicarboxylate zwitterions and three nonzwitterionic methoxycarbonylpyridine carboxylate isomers (-O(2)CPyrCO(2)CH(3)). Information about the intramolecular electrostatic interactions was directly obtained from the photoelectron spectra. The adiabatic and vertical detachment energies were measured and understood in terms of intramolecular Coulombic forces. Calculations at the B3LYP and CCSD(T) level were performed and compared to the experimental electron binding energies.more » Structures, relative stabilities, and the electron detachment sites also were obtained from the calculations.« less

  6. 40 CFR 417.170 - Applicability; description of the manufacture of detergents by dry blending subcategory.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... manufacture of detergents by dry blending subcategory. 417.170 Section 417.170 Protection of Environment... POINT SOURCE CATEGORY Manufacture of Detergents by Dry Blending Subcategory § 417.170 Applicability; description of the manufacture of detergents by dry blending subcategory. The provisions of this subpart are...

  7. 40 CFR 417.190 - Applicability; description of the manufacture of detergent bars and cakes subcategory.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... manufacture of detergent bars and cakes subcategory. 417.190 Section 417.190 Protection of Environment... POINT SOURCE CATEGORY Manufacture of Detergent Bars and Cakes Subcategory § 417.190 Applicability; description of the manufacture of detergent bars and cakes subcategory. The provisions of this subpart are...

  8. The Complementary Health Approaches for Pain Survey (CHAPS): Validity testing and characteristics of a rural population with pain

    PubMed Central

    2018-01-01

    Objectives Little is known about patterns and correlates of Complementary Health Approaches (CHAs) in chronic pain populations, particularly in rural, underserved communities. This article details the development and implementation of a new survey instrument designed to address this gap, the Complementary Health Approaches for Pain Survey (CHAPS). Design Following pilot-testing using pre-specified criteria to assess quality and comprehension in our target population, and after feedback regarding face-validity from content experts and stakeholders, the final cross-sectional self-report survey required 10–12 minutes to complete. It contained 69 demographic, lifestyle and health-related factors, and utilized a Transtheoretical Model (TTM) underpinning to assess short- and long-term use of 12 CHAs for pain management. Twenty additional items on pain severity, feelings, clinical outcomes, and activities were assessed using the Short-Form Global Pain Scale (SF-GPS); Internal reliability was assessed using Cronbach’s alpha. Settings/location Investigators conducted consecutive sampling in four West Virginia pain management and rheumatology practices. Participants 301 Appalachian adult patients seeking conventional care for pain management. Results Response rates were high (88% ± 4.1%). High quality and comprehension deemed the CHAPS an appropriate measurement tool in a rural population with pain. Missing data were unrelated to patient characteristics. Participants predominantly experienced chronic pain (93%), had five or more health conditions (56%, Mean = 5.4±3.1), were white (92%), female (57%), and middle-aged (Mean = 55.6 (SD = 13.6) years). Over 40% were disabled (43%) and/or obese (44%, Mean BMI = 33.4±31.5). Additionally, 44% used opioids, 31% used other prescription medications, and 66% used at least one CHA for pain, with 48% using CHAs for greater than 6 months. There was high internal reliability of the SF-GPS (alpha = .93) and satisfactory internal

  9. Detergent alkaline proteases: enzymatic properties, genes, and crystal structures.

    PubMed

    Saeki, Katsuhisa; Ozaki, Katsuya; Kobayashi, Tohru; Ito, Susumu

    2007-06-01

    Subtilisin-like serine proteases from bacilli have been used in various industrial fields worldwide, particularly in the production of laundry and automatic dishwashing detergents. They belong to family A of the subtilase superfamily, which is composed of three clans, namely, true subtilisins, high-alkaline proteases, and intracellular proteases. We succeeded in the large-scale production of a high-alkaline protease (M-protease) from alkaliphilic Bacillus clausii KSM-K16, and the enzyme has been introduced into compact heavy-duty laundry detergents. We have also succeeded in the industrial-scale production of a new alkaline protease, KP-43, which was originally resistant to chemical oxidants and to surfactants, produced by alkaliphilic Bacillus sp. strain KSM-KP43 and have incorporated it into laundry detergents. KP-43 and related proteases form a new clan, oxidatively stable proteases, in subtilase family A. In this review, we describe the enzymatic properties, gene sequences, and crystal structures of M-protease, KP-43, and related enzymes.

  10. Structure formation in binary mixtures of lipids and detergents: self-assembly and vesicle division.

    PubMed

    Noguchi, Hiroshi

    2013-01-14

    Self-assembly dynamics in binary surfactant mixtures and structure changes of lipid vesicles induced by detergent solution are studied using coarse-grained molecular simulations. Disk-shaped micelles, the bicelles, are stabilized by detergents surrounding the rim of a bilayer disk of lipids. The self-assembled bicelles are considerably smaller than bicelles formed from vesicle rupture, and their size is determined by the concentrations of lipids and detergents and the interactions between the two species. The detergent-adsorption induces spontaneous curvature of the vesicle bilayer and results in vesicle division into two vesicles or vesicle rupture into worm-like micelles. The division occurs mainly via the inverse pathway of the modified stalk model. For large spontaneous curvature of the monolayers of the detergents, a pore is often opened, thereby leading to vesicle division or worm-like micelle formation.

  11. Zwitterionic sulfobetaine polymer-immobilized surface by simple tyrosinase-mediated grafting for enhanced antifouling property.

    PubMed

    Kwon, Ho Joon; Lee, Yunki; Phuong, Le Thi; Seon, Gyeung Mi; Kim, Eunsuk; Park, Jong Chul; Yoon, Hyunjin; Park, Ki Dong

    2017-10-01

    Introducing antifouling property to biomaterial surfaces has been considered an effective method for preventing the failure of implanted devices. In order to achieve this, the immobilization of zwitterions on biomaterial surfaces has been proven to be an excellent way of improving anti-adhesive potency. In this study, poly(sulfobetaine-co-tyramine), a tyramine-conjugated sulfobetaine polymer, was synthesized and simply grafted onto the surface of polyurethane via a tyrosinase-mediated reaction. Surface characterization by water contact angle measurements, X-ray photoelectron spectroscopy and atomic force microscopy demonstrated that the zwitterionic polymer was successfully introduced onto the surface of polyurethane and remained stable for 7days. In vitro studies revealed that poly(sulfobetaine-co-tyramine)-coated surfaces dramatically reduced the adhesion of fibrinogen, platelets, fibroblasts, and S. aureus by over 90% in comparison with bare surfaces. These results proved that polyurethane surfaces grafted with poly(sulfobetaine-co-tyramine) via a tyrosinase-catalyzed reaction could be promising candidates for an implantable medical device with excellent bioinert abilities. Antifouling surface modification is one of the key strategy to prevent the thrombus formation or infection which occurs on the surface of biomaterial after transplantation. Although there are many methods to modify the surface have been reported, necessity of simple modification technique still exists to apply for practical applications. The purpose of this study is to modify the biomaterial's surface by simply immobilizing antifouling zwitterion polymer via enzyme tyrosinase-mediated reaction which could modify versatile substrates in mild aqueous condition within fast time period. After modification, pSBTA grafted surface becomes resistant to various biological factors including proteins, cells, and bacterias. This approach appears to be a promising method to impart antifouling property on

  12. Enhancing MRI of liver metastases with a zwitterionized biodegradable dendritic contrast agent.

    PubMed

    Zhou, Xiaoxuan; Ye, Mingzhou; Han, Yuxin; Tang, Jianbin; Qian, Yue; Hu, Hongjie; Shen, Youqing

    2017-07-25

    Metastasis is the main reason for cancer-associated mortality, and accurate diagnostic imaging of metastases is critical for the clinical administration and tailoring personalized treatments for metastatic tumors. However, magnetic resonance imaging of metastases in the liver is impeded by its low sensitivity because the currently used contrast agents accumulate in hepatocytes and Kupffer cells instead of cancer cells. Herein, a 4 th generation zwitterionized biodegradable dendritic contrast agent (DCA) with a size of ca. 9 nm and a longitudinal relaxivity of 15.7 mM -1 s -1 in terms of Gd was synthesized and used to enhance the MRI of liver metastasis. The DCA could remarkably enhance the MRI of metastasized tumors in the liver, because it could simultaneously reduce the background signal in the liver by avoiding uptake by hepatocytes and Kupffer cells through the zwitterionization and increase the signal in tumors through the enhanced permeability and retention effect. Moreover, in contrast to non-biodegradable DCA, this DCA showed minimal long-term Gd 3+ retention in all organs and tissues because it could be degraded into small fragments. The significant capability of enhancing the MRI of metastases in the liver plus its excellent biodegradability made this DCA a promising CA for metastatic tumor imaging.

  13. Detergents modify proteinase K resistance of PrPSc in different transmissible spongiform encephalopathies (TSEs)

    PubMed Central

    Breyer, Johanna; Wemheuer, Wiebke M.; Wrede, Arne; Graham, Catherine; Benestad, Sylvie L.; Brenig, Bertram; Richt, Jürgen A.; Schulz-Schaeffer, Walter J.

    2012-01-01

    Prion diseases are diagnosed by the detection of their proteinase K-resistant prion protein fragment (PrPSc). Various biochemical protocols use different detergents for the tissue preparation. We found that the resistance of PrPSc against proteinase K may vary strongly with the detergent used. In our study, we investigated the influence of the most commonly used detergents on eight different TSE agents derived from different species and distinct prion disease forms. For a high throughput we used a membrane adsorbtion assay to detect small amounts of prion aggregates, as well as Western blotting. Tissue lysates were prepared using DOC, SLS, SDS or Triton X-100 in different concentrations and these were digested with various amounts of proteinase K. Detergents are able to enhance or diminish the detectability of PrPSc after proteinase K digestion. Depending on the kind of detergent, its concentration - but also on the host species that developed the TSE and the disease form or prion type - the detectability of PrPSc can be very different. The results obtained here may be helpful during the development or improvement of a PrPSc detection method and they point towards a detergent effect that can be additionally used for decontamination purposes. A plausible explanation for the detergent effects described in this article could be an interaction with the lipids associated with PrPSc that may stabilize the aggregates. PMID:22226540

  14. 40 CFR 80.156 - Liability for violations of the interim detergent program controls and prohibitions.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... base gasoline component, the detergent component, or the detergent-additized post-refinery component of... component of any post-refinery component or gasoline in the storage tank containing gasoline found to be in... evidence, that the gasoline or detergent carrier caused the violation. (2) Post-refinery component non...

  15. False-positive pregnancy test after transfusion of solvent/detergent-treated plasma.

    PubMed

    Jilma-Stohlawetz, Petra; Wreford-Bush, Tim; Mills, Francesca; Davidson, Fiona; Kursten, Friedrich W; Jilma, Bernd; Birchall, Janet

    2017-12-01

    The transmission of pathogens, antibodies, and proteins is a possible consequence of blood product transfusion. A female patient had an unexpected positive serum β-human chorionic gonadotropin result, indicative of pregnancy, after she had received a transfusion with 1 unit of platelet concentrate, 4 units of red blood cells, and 4 units of pooled solvent/detergent-treated plasma (Octaplas). To investigate the possibility of passive transfusion of β-human chorionic gonadotropin from the plasma transfusion, one additional unit from the same batch was thawed and analyzed. To validate the β-human chorionic gonadotropin assay for use in solvent/detergent-treated plasma and to investigate any interference in the assay, dilution experiments were performed using the implicated plasma batch diluted with male and non-pregnant female sera. Also, plasma from a known pregnant woman was diluted with Octaplas (tested negative for β-human chorionic gonadotropin) and with a male serum to validate the assay for use in solvent/detergent-treated plasma. The implicated solvent/detergent-treated plasma had a mean β-human chorionic gonadotropin level of 91.5 mIU/mL. Results from the dilution experiments revealed an excellent correlation (r > 0.99) between β-human chorionic gonadotropin measurement in solvent/detergent-treated plasma and male serum and no over or under recovery of the expected results. Further measurements of β-human chorionic gonadotropin levels in the female recipient revealed an estimated half-life of 6 hours. This case demonstrates the importance of considering the possibility of passive transmission of analytes to a patient from the transfusion of blood products. Furthermore, the measurement of β-human chorionic gonadotropin is valid in solvent/detergent-treated plasma using a Roche Cobas analyzer. © 2017 AABB.

  16. Occupational asthma caused by cellulase and lipase in the detergent industry.

    PubMed

    Brant, A; Hole, A; Cannon, J; Helm, J; Swales, C; Welch, J; Taylor, A Newman; Cullinan, P

    2004-09-01

    Three employees from two different detergent companies were investigated for occupational asthma, using skin prick tests, serum specific IgE, and specific bronchial challenge. Two were challenged with lipase and one with cellulase. All three cases had immunological evidence of sensitisation to the detergent enzymes with which they worked. Bronchial challenge in each provoked a reproducible dual asthmatic response, which reproduced their work related symptoms. These are the first reported cases of occupational asthma attributable to cellulase and lipase in the detergent industry. Four of the most common enzymes used in this industry have now been reported to cause occupational asthma; continued vigilance and caution are needed when working with these or other enzymes.

  17. Efficacy of detergent and water versus bleach for disinfection of direct contact ophthalmic lenses.

    PubMed

    Abbey, Ashkan M; Gregori, Ninel Z; Surapaneni, Krishna; Miller, Darlene

    2014-06-01

    Although manufacturers recommend cleaning ophthalmic lenses with detergent and water and then with a specific disinfectant, disinfectants are rarely used in ophthalmic practices. The aim of this pilot study was to evaluate the efficacy of detergent and water versus that of bleach, a recommended disinfectant, to eliminate common ocular bacteria and viruses from ophthalmic lenses. Three bacterial strains (Staphylococcus epidermidis, Corynebacterium straitum, and methicillin-resistant Staphylococcus aureus and 2 viral strains (adenovirus and herpes simplex virus [HSV] type-1) were individually inoculated onto 20 gonioscopy and laser lenses. The lenses were washed with detergent and water and then disinfected with 10% bleach. All the lenses were cultured after inoculation, after washing with detergent and water, and after disinfecting with the bleach. Bacterial cultures in thioglycollate broth were observed for 3 weeks, and viral cultures were observed for 2 weeks. The presence of viruses was also detected by multiplex polymerase chain reaction (PCR). All 20 lenses inoculated with S. epidermidis, C. straitum, adenovirus, and HSV-1 showed growth after inoculation but no growth after washing with detergent/water and after disinfecting with the bleach. All lenses showed positive HSV and adenovirus PCR results after inoculation and negative PCR results after washing with detergent/water and after disinfecting with bleach. All methicillin-resistant S. aureus-contaminated lenses showed growth after inoculation and no growth after washing with detergent and water. However, 1 lens showed positive growth after disinfecting with bleach. Cleaning with detergent and water seemed to effectively eliminate bacteria and viruses from the surface of contaminated ophthalmic lenses. Further studies are warranted to design practical disinfection protocols that minimize lens damage.

  18. Characterization of the Activity and Stability of Amylase from Saliva and Detergent: Laboratory Practicals for Studying the Activity and Stability of Amylase from Saliva and Various Commercial Detergents

    ERIC Educational Resources Information Center

    Valls, Cristina; Rojas, Cristina; Pujadas, Gerard; Garcia-Vallve, Santi; Mulero, Miquel

    2012-01-01

    This article presents two integrated laboratory exercises intended to show students the role of [alpha]-amylases (AAMYs) in saliva and detergents. These laboratory practicals are based on the determination of the enzymatic activity of amylase from saliva and different detergents using the Phadebas test (quantitative) and the Lugol test…

  19. Conjugated Polymer Zwitterions: Efficient Interlayer Materials in Organic Electronics.

    PubMed

    Liu, Yao; Duzhko, Volodimyr V; Page, Zachariah A; Emrick, Todd; Russell, Thomas P

    2016-11-15

    Conjugated polymer zwitterions (CPZs) are neutral, hydrophilic, polymer semiconductors. The pendent zwitterions, viewed as side chain dipoles, impart solubility in polar solvents for solution processing, and open opportunities as interfacial components of optoelectronic devices, for example, between metal electrodes and organic semiconductor active layers. Such interlayers are crucial for defining the performance of organic electronic devices, e.g., field-effect transistors (OFETs), light-emitting diodes (OLEDs), and photovoltaics (OPVs), all of which consist of multilayer structures. The interlayers reduce the Schottky barrier height and thus improve charge injection in OFETs and OLEDs. In OPVs, the interlayers serve to increase the built-in electric potential difference (V bi ) across the active layer, ensuring efficient extraction of photogenerated charge carriers. In general, polar and even charged electronically active polymers have gained recognition for their ability to modify metal/semiconductor interfaces to the benefit of organic electronics. While conjugated polyelectrolytes (CPEs) as interlayer materials are well-documented, open questions remain about the role of mobile counterions in CPE-containing devices. CPZs possess the processing advantages of CPEs, but as neutral molecules lack any potential complications associated with counterions. The electronic implications of CPZs on metal electrodes stem from the orientation of the zwitterion dipole moment in close proximity to the metal surface, and the resultant surface-induced polarization. This generates an interfacial dipole (Δ) at the CPZ/metal interface, altering the work function of the electrode, as confirmed by ultraviolet photoelectron spectroscopy (UPS), and improving device performance. An ideal cathode interlayer would reduce electrode work function, have orthogonal processability to the active layer, exhibit good film forming properties (i.e., wettability/uniformity), prevent exciton

  20. Submicrometer Emitter ESI Tips for Native Mass Spectrometry of Membrane Proteins in Ionic and Nonionic Detergents

    NASA Astrophysics Data System (ADS)

    Susa, Anna C.; Lippens, Jennifer L.; Xia, Zijie; Loo, Joseph A.; Campuzano, Iain D. G.; Williams, Evan R.

    2018-01-01

    Native mass spectrometry (native-MS) of membrane proteins typically requires a detergent screening protocol, protein solubilization in the preferred detergent, followed by protein liberation from the micelle by collisional activation. Here, submicrometer nano-ESI emitter tips are used for native-MS of membrane proteins solubilized in both nonionic and ionic detergent solutions. With the submicrometer nano-ESI emitter tips, resolved charge-state distributions of membrane protein ions are obtained from a 150 mM NaCl, 25 mM Tris-HCl with 1.1% octyl glucoside solution. The relative abundances of NaCl and detergent cluster ions at high m / z are significantly reduced with the submicrometer emitters compared with larger nano-ESI emitters that are commonly used. This technique is beneficial for significantly decreasing the abundances (by two to three orders of magnitude compared with the larger tip size: 1.6 μm) of detergent cluster ions formed from aqueous ammonium acetate solutions containing detergents that can overlap with the membrane protein ion signal. Resolved charge-state distributions of membrane protein ions from aqueous ammonium acetate solutions containing ionic detergents were obtained with the submicrometer nano-ESI emitters; this is the first report of native-MS of membrane proteins solubilized by ionic detergents. [Figure not available: see fulltext.

  1. Submicrometer Emitter ESI Tips for Native Mass Spectrometry of Membrane Proteins in Ionic and Nonionic Detergents.

    PubMed

    Susa, Anna C; Lippens, Jennifer L; Xia, Zijie; Loo, Joseph A; Campuzano, Iain D G; Williams, Evan R

    2018-01-01

    Native mass spectrometry (native-MS) of membrane proteins typically requires a detergent screening protocol, protein solubilization in the preferred detergent, followed by protein liberation from the micelle by collisional activation. Here, submicrometer nano-ESI emitter tips are used for native-MS of membrane proteins solubilized in both nonionic and ionic detergent solutions. With the submicrometer nano-ESI emitter tips, resolved charge-state distributions of membrane protein ions are obtained from a 150 mM NaCl, 25 mM Tris-HCl with 1.1% octyl glucoside solution. The relative abundances of NaCl and detergent cluster ions at high m /z are significantly reduced with the submicrometer emitters compared with larger nano-ESI emitters that are commonly used. This technique is beneficial for significantly decreasing the abundances (by two to three orders of magnitude compared with the larger tip size: 1.6 μm) of detergent cluster ions formed from aqueous ammonium acetate solutions containing detergents that can overlap with the membrane protein ion signal. Resolved charge-state distributions of membrane protein ions from aqueous ammonium acetate solutions containing ionic detergents were obtained with the submicrometer nano-ESI emitters; this is the first report of native-MS of membrane proteins solubilized by ionic detergents. Graphical Abstract.

  2. Detergent-Mediated Formation of β-Hematin: Heme Crystallization Promoted by Detergents Implicates Nanostructure Formation for Use as a Biological Mimic

    PubMed Central

    2016-01-01

    Hemozoin is a unique biomineral that results from the sequestration of toxic free heme liberated as a consequence of hemoglobin degradation in the malaria parasite. Synthetic neutral lipid droplets (SNLDs) and phospholipids were previously shown to support the rapid formation of β-hematin, abiological hemozoin, under physiologically relevant pH and temperature, though the mechanism by which heme crystallization occurs remains unclear. Detergents are particularly interesting as a template because they are amphiphilic molecules that spontaneously organize into nanostructures and have been previously shown to mediate β-hematin formation. Here, 11 detergents were investigated to elucidate the physicochemical properties that best recapitulate crystal formation in the parasite. A strong correlation between the detergent’s molecular structure and the corresponding kinetics of β-hematin formation was observed, where higher molecular weight polar chains promoted faster reactions. The larger hydrophilic chains correlated to the detergent’s ability to rapidly sequester heme into the lipophilic core, allowing for crystal nucleation to occur. The data presented here suggest that detergent nanostructures promote β-hematin formation in a similar manner to SNLDs and phospholipids. Through understanding mediator properties that promote optimal crystal formation, we are able to establish an in vitro assay to probe this drug target pathway. PMID:27175104

  3. Zwitterionic sulfobetaine-grafted poly(vinylidene fluoride) membrane with highly effective blood compatibility via atmospheric plasma-induced surface copolymerization.

    PubMed

    Chang, Yung; Chang, Wan-Ju; Shih, Yu-Ju; Wei, Ta-Chin; Hsiue, Ging-Ho

    2011-04-01

    Development of nonfouling membranes to prevent nonspecific protein adsorption and platelet adhesion is critical for many biomedical applications. It is always a challenge to control the surface graft copolymerization of a highly polar monomer from the highly hydrophobic surface of a fluoropolymer membrane. In this work, the blood compatibility of poly(vinylidene fluoride) (PVDF) membranes with surface-grafted electrically neutral zwitterionic poly(sulfobetaine methacrylate) (PSBMA), from atmospheric plasma-induced surface copolymerization, was studied. The effect of surface composition and graft morphology, electrical neutrality, hydrophilicity and hydration capability on blood compatibility of the membranes were determined. Blood compatibility of the zwitterionic PVDF membranes was systematically evaluated by plasma protein adsorption, platelet adhesion, plasma-clotting time, and blood cell hemolysis. It was found that the nonfouling nature and hydration capability of grafted PSBMA polymers can be effectively controlled by regulating the grafting coverage and charge balance of the PSBMA layer on the PVDF membrane surface. Even a slight charge bias in the grafted zwitterionic PSBMA layer can induce electrostatic interactions between proteins and the membrane surfaces, leading to surface protein adsorption, platelet activation, plasma clotting and blood cell hemolysis. Thus, the optimized PSBMA surface graft layer in overall charge neutrality has a high hydration capability and the best antifouling, anticoagulant, and antihemolytic activities when comes into contact with human blood. © 2011 American Chemical Society

  4. Properties of Zeolite A Obtained from Powdered Laundry Detergent: An Undergraduate Experiment.

    ERIC Educational Resources Information Center

    Smoot, Alison L.; Lindquist, David A.

    1997-01-01

    Presents experiments that introduce students to the myriad properties of zeolites using the sodium form of zeolite A (Na-A) from laundry detergent. Experiments include extracting Na-A from detergent, water softening properties, desiccant properties, ion-exchange properties, and Zeolite HA as a dehydration catalyst. (JRH)

  5. Study on OELs for enzyme-containing detergent in China.

    PubMed

    Zhang, X D; Liang, Y X; Lee, C S; Jin, T Y

    2004-01-01

    This study is aimed at setting occupational exposure levels for total detergent dust and enzymes in detergent industries. The study population consisted of 795 workers from four enzyme-containing detergent manufacturing plants (A1, A2, B1 and B2), and 156 control workers from an electronic assembly factory. Work environment monitoring was conducted using high volume of air sampler fro measuring the concentration of total dust (mg/m3), and analyzing the level of enzyme (ng/m3) by ELISA method. A standard questionnaires, pulmonary function test, and skin prick test are used to assess health effects. The levels of detergent total dust varied from 0.2 mg/m3 to 12.54 mg/m3. For enzyme levels, in A1, B1 and B2, the concentration ranged from non-detectable to 9.92 ng/m3 and in A2, the concentration was analyzed by enzyme activity methods and was expressed as Gu/m3 (1 Gu/m3 = 16 ng/m3). The concentration is between 0.16-31.36 ng/m3. Non-specific irritation rates in exposed workers were significantly higher than that in controls. Based on the data collected from A1, B1 and control plants, 95% benchmark dose lower bound were calculated as 1.17 mg/m3. The difference of pulmonary function between exposed workers and controls is not significant. The results of SPT showed that neither Savinase- nor Alcalase-induced sensitization was found in controls. The prevalence rates of sensitization for Savinase and Alcalase were ranged between 3.2% and 31% in all enzyme-containing detergent manufacturers investigated. No case of occupational asthma was observed. For total dust, 1 mg/m3 is suggested as permissible concentration-time weighted average (PC-TWA), and 2 mg/m3 as permissible concentration-short term exposure limit (PC-STEL). For the enzyme Subtilisins, 15 ng/m3 is suggested as PC-TWA, and 30 ng/m3 as PC-STEL.

  6. Systematic analysis of protein–detergent complexes applying dynamic light scattering to optimize solutions for crystallization trials

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

    Meyer, Arne; Dierks, Karsten; XtalConcepts, Marlowring 19, 22525 Hamburg

    Application of in situ dynamic light scattering to solutions of protein–detergent complexes permits characterization of these complexes in samples as small as 2 µl in volume. Detergents are widely used for the isolation and solubilization of membrane proteins to support crystallization and structure determination. Detergents are amphiphilic molecules that form micelles once the characteristic critical micelle concentration (CMC) is achieved and can solubilize membrane proteins by the formation of micelles around them. The results are presented of a study of micelle formation observed by in situ dynamic light-scattering (DLS) analyses performed on selected detergent solutions using a newly designed advancedmore » hardware device. DLS was initially applied in situ to detergent samples with a total volume of approximately 2 µl. When measured with DLS, pure detergents show a monodisperse radial distribution in water at concentrations exceeding the CMC. A series of all-transn-alkyl-β-d-maltopyranosides, from n-hexyl to n-tetradecyl, were used in the investigations. The results obtained verify that the application of DLS in situ is capable of distinguishing differences in the hydrodynamic radii of micelles formed by detergents differing in length by only a single CH{sub 2} group in their aliphatic tails. Subsequently, DLS was applied to investigate the distribution of hydrodynamic radii of membrane proteins and selected water-insoluble proteins in presence of detergent micelles. The results confirm that stable protein–detergent complexes were prepared for (i) bacteriorhodopsin and (ii) FetA in complex with a ligand as examples of transmembrane proteins. A fusion of maltose-binding protein and the Duck hepatitis B virus X protein was added to this investigation as an example of a non-membrane-associated protein with low water solubility. The increased solubility of this protein in the presence of detergent could be monitored, as well as the progress of

  7. High-Melting Lipid Mixtures and the Origin of Detergent-Resistant Membranes Studied with Temperature-Solubilization Diagrams

    PubMed Central

    Sot, Jesús; Manni, Marco M.; Viguera, Ana R.; Castañeda, Verónica; Cano, Ainara; Alonso, Cristina; Gil, David; Valle, Mikel; Alonso, Alicia; Goñi, Félix M.

    2014-01-01

    The origin of resistance to detergent solubilization in certain membranes, or membrane components, is not clearly understood. We have studied the solubilization by Triton X-100 of binary mixtures composed of egg sphingomyelin (SM) and either ceramide, diacylglycerol, or cholesterol. Solubilization has been assayed in the 4–50°C range, and the results are summarized in a novel, to our knowledge, form of plots, that we have called temperature-solubilization diagrams. Despite using a large detergent excess (lipid/detergent 1:20 mol ratio) and extended solubilization times (24–48 h) certain mixtures were not amenable to Triton X-100 solubilization at one or more temperatures. DSC of all the lipid mixtures, and of all the lipid + detergent mixtures revealed that detergent resistance was associated with the presence of gel domains at the assay temperature. Once the system melted down, solubilization could occur. In general adding high-melting lipids limited the solubilization, whereas the addition of low-melting lipids promoted it. Lipidomic analysis of Madin-Darby canine kidney cell membranes and of the corresponding detergent-resistant fraction indicated a large enrichment of the nonsolubilized components in saturated diacylglycerol and ceramide. SM-cholesterol mixtures were special in that detergent solubilization was accompanied, for certain temperatures and compositions, by an independent phenomenon of reassembly of the partially solubilized lipid bilayers. The temperature at which lysis and reassembly prevailed was ∼25°C, thus for some SM-cholesterol mixtures solubilization occurred both above and below 25°C, but not at that temperature. These observations can be at the origin of the detergent resistance effects observed with cell membranes, and they also mean that cholesterol-containing detergent-resistant membrane remnants cannot correspond to structures existing in the native membrane before detergent addition. PMID:25517149

  8. [Status of the upper respiratory tract of workers in the synthetic detergent industry].

    PubMed

    Artamonova, V G; Barsukov, A F; Galzhiev, A S

    1991-01-01

    Otorhinolaryngologic examinations and functional studies were performed among 99 workers engaged in the production of synthetic detergents. The studies revealed disorders in the barrier and decontamination functions of the nasal ciliary epithelium caused by synthetic detergent dusts.

  9. Efficacy of Detergent and Water Versus Bleach for the Disinfection of Direct Contact Ophthalmic Lenses

    PubMed Central

    Abbey, Ashkan M.; Gregori, Ninel Z.; Surapaneni, Krishna; Miller, Darlene

    2014-01-01

    Purpose While manufacturers recommend cleaning ophthalmic lenses with detergent and water and then a specific disinfectant, disinfectants are rarely used in ophthalmic practices. The aim of this pilot study was to evaluate the efficacy of detergent and water versus bleach, a recommended disinfectant, to eliminate common ocular bacteria and viruses from ophthalmic lenses. Methods Three bacterial strains (Staphylococcus epidermidis, Corynebacterium straitum, and methicillin-resistant Staphylococcus aureus (MRSA) and two viral strains (adenovirus and herpes simplex virus (HSV) type-1) were individually inoculated to 20 gonioscopy and laser lenses. Lenses were washed with detergent and water and then disinfected with 10% bleach. All lenses were cultured after inoculation, after detergent and water, and after the bleach. Bacterial cultures in thioglycollate broth were observed for 3 weeks and viral cultures for 2 weeks. The presence of viruses was also detected by multiplex polymerase chain reaction (PCR). Results All 20 lenses inoculated with Staphylococcus epidermidis, Corynebacterium straitum, adenovirus, and HSV-1 showed growth after inoculation, but no growth after detergent/water and after the bleach. All lenses showed positive HSV and adenovirus PCR after inoculation and negative PCR after detergent/water and after bleach. All MRSA contaminated lenses showed growth after inoculation and no growth after detergent and water. However, one lens showed positive growth after bleach. Conclusions Cleaning with detergent and water appeared to effectively eliminate bacteria and viruses from the surface of contaminated ophthalmic lenses. Further studies are warranted to design practical disinfection protocols that minimize lens damage. PMID:24747806

  10. Self-Assembling Peptide Detergents Stabilize Isolated Photosystem Ion a Dry Surface for an Extended Time

    PubMed Central

    Kiley, Patrick; Zhao, Xiaojun; Vaughn, Michael; Baldo, Marc A; Bruce, Barry D

    2005-01-01

    We used a class of designed peptide detergents to stabilize photosystem I (PS-I) upon extended drying under N2 on a gold-coated-Ni-NTA glass surface. PS-I is a chlorophyll-containing membrane protein complex that is the primary reducer of ferredoxin and the electron acceptor of plastocyanin. We isolated the complex from the thylakoids of spinach chloroplasts using a chemical detergent. The chlorophyll molecules associated with the PS-I complex provide an intrinsic steady-state emission spectrum between 650 and 800 nm at −196.15 °C that reflects the organization of the pigment-protein interactions. In the absence of detergents, a large blue shift of the fluorescence maxima from approximately 735 nm to approximately 685 nm indicates a disruption in light-harvesting subunit organization, thus revealing chlorophyll−protein interactions. The commonly used membrane protein-stabilizing detergents, N-dodecyl-β-D-maltoside and N-octyl-β-D-glucoside, only partially stabilized the approximately 735-nm complex with approximately 685-nm spectroscopic shift. However, prior to drying, addition of the peptide detergent acetyl- AAAAAAK at increasing concentration significantly stabilized the PS-I complex. Moreover, in the presence of acetyl- AAAAAAK, the PS-I complex is stable in a dried form at room temperature for at least 3 wk. Another peptide detergent, acetyl-VVVVVVD, also stabilized the complex but to a lesser extent. These observations suggest that the peptide detergents may effectively stabilize membrane proteins in the solid-state. These designed peptide detergents may facilitate the study of diverse types of membrane proteins. PMID:15954800

  11. Aerosol Chemical Composition and its Effects on Cloud-Aerosol Interactions during the 2007 CHAPS Experiment

    NASA Astrophysics Data System (ADS)

    Lee, Y.; Alexander, L.; Newburn, M.; Jayne, J.; Hubbe, J.; Springston, S.; Senum, G.; Andrews, B.; Ogren, J.; Kleinman, L.; Daum, P.; Berg, L.; Berkowitz, C.

    2007-12-01

    Chemical composition of submicron aerosol particles was determined using an Aerodyne Time-of-Flight Aerosol Mass Spectrometer (AMS) outfitted on the DOE G-1 aircraft during the Cumulus Humilis Aerosol Processing Study (CHAPS) conducted in Oklahoma City area in June 2007. The primary objective of CHAPS was to investigate the effects of urban emissions on cloud aerosol interactions as a function of processing of the emissions. Aerosol composition was typically determined at three different altitudes: below, in, and above cloud, in both upwind and downwind regions of the urban area. Aerosols were sampled from an isokinetic inlet with an upper size cut-off of ~1.5 micrometer. During cloud passages, the AMS also sampled particles that were dried from cloud droplets collected using a counter-flow virtual impactor (CVI) sampler. The aerosol mass concentrations were typically below 10 microgram per cubic meter, and were dominated by organics and sulfate. Ammonium was often less than required for complete neutralization of sulfate. Aerosol nitrate levels were very low. We noted that nitrate levels were significantly enhanced in cloud droplets compared to aerosols, most likely resulting from dissolution of gaseous nitric acid. Organic to sulfate ratios appeared to be lower in cloud droplets than in aerosols, suggesting cloud condensation nuclei properties of aerosol particles might be affected by loading and nature of the organic components in aerosols. In-cloud formation of sulfate was considered unimportant because of the very low SO2 concentration in the region. A detailed examination of the sources of the aerosol organic components (based on hydrocarbons determined using a proton transfer reaction mass spectrometer) and their effects on cloud formation as a function of atmospheric processing (based on the degree of oxidation of the organic components) will be presented.

  12. Sublethal detergent concentrations increase metabolization of recalcitrant polyphosphonates by the cyanobacterium Spirulina platensis.

    PubMed

    Forlani, Giuseppe; Bertazzini, Michele; Giberti, Samuele; Wieczorek, Dorota; Kafarski, Paweł; Lipok, Jacek

    2013-05-01

    As a consequence of increasing industrial applications, thousand tons of polyphosphonates are introduced every year into the environment. The inherent stability of the C-P bond results in a prolonged half-life. Moreover, low uptake rates limit further their microbial metabolization. To assess whether low detergent concentrations were able to increase polyphosphonate utilization by the cyanobacterium Spirulina platensis, tolerance limits to the exposure to various detergents were determined by measuring the growth rate in the presence of graded levels below the critical micellar concentration. Then, the amount of hexamethylenediamine-N,N,N',N'-tetrakis(methylphosphonic acid) that is metabolized in the absence or in the presence of sublethal detergent concentrations was quantified by (31)P NMR analysis on either P-starved or P-fed cyanobacterial cultures. The strain tolerated the presence of detergents in the order: nonionic > anionic > cationic. When added to the culture medium at the highest concentrations showing no detrimental effects upon cell viability, detergents either improved or decreased polyphosphonate utilization, the anionic sodium dodecyl sulfate being the most beneficial. Metabolization was not lower in P-fed cells--a result that strengthens the possibility of using, in the future, this strain for bioremediation purposes.

  13. Effect of carbon spacer length on zwitterionic carboxybetaines.

    PubMed

    Shao, Qing; Jiang, Shaoyi

    2013-02-07

    Zwitterionic carboxybetaines (CBs) are ubiquitous in nature and considered promising materials for biological and chemical applications. A thorough understanding of the effect of carbon spacer length (CSL) on molecular properties is important. In this work, using molecular dynamics simulation and quantum chemical calculation, we investigated the effect of CSL on the molecular properties of CB molecules. The hydration number, structure, and dynamics of carboxylic and trimethyl ammonium groups were investigated and found to present different behaviors in regards to the variation of CSL. The simulation results with partial charges developed from quantum chemical calculations were compared with those with partial charges from the OPLS all atom (OPLSAA) force field. The hydration free energy of CB molecules and CB-Na(+) association was also studied as a function of CSL.

  14. Effect of synthetic detergents on the swelling and the ATPase of mitochondria isolated from rat liver.

    PubMed

    WITTER, R F; MINK, W

    1958-01-25

    A study was made of the effects of various types of detergents on the swelling of isolated mitochondria and on mitochondrial ATPases which are activated by Mg or DNP respectively. The rate of swelling was measured in the Beckman spectrophotometer by following the decrease in turbidity of dilute suspensions of these organelles. It was found that non-ionic detergents containing a nonyl phenoxy side chain or anionic detergents caused swelling of the mitochondria and activation of Mg-ATPase. On the other hand, cationic detergents promoted the clumping of mitochondria and did not activate Mg-ATPase. DNP-ATPase was inhibited by all of the detergents tested. It would appear from these observations that the inhibition of DNP-ATPase is not related to a gross change in the morphology of the organelles; in contrast, the activation of Mg-ATPase definitely is correlated with swelling of the isolated mitochondria. These data also suggest that the ionic detergents combine with charged sites on the protein moiety of the lipoprotein in the mitochondrial surface, whereas the non-ionic detergents form inclusion compounds with the lipide moiety, thereby altering the mitochondrial structure and permeability.

  15. The variable detergent sensitivity of proteases that are utilized for recombinant protein affinity tag removal

    PubMed Central

    Vergis, James M.; Wiener, Michael C.

    2011-01-01

    Recombinant proteins typically include one or more affinity tags to facilitate purification and/or detection. Expression constructs with affinity tags often include an engineered protease site for tag removal. Like other enzymes, the activities of proteases can be affected by buffer conditions. The buffers used for integral membrane proteins contain detergents, which are required to maintain protein solubility. We examined the detergent sensitivity of six commonly-used proteases (Enterokinase, Factor Xa, Human Rhinovirus 3C Protease, SUMOstar, Tobacco Etch Virus Protease, and Thrombin) by use of a panel of ninety-four individual detergents. Thrombin activity was insensitive to the entire panel of detergents, thus suggesting it as the optimal choice for use with membrane proteins. Enterokinase and Factor Xa were only affected by a small number of detergents, making them good choices as well. PMID:21539919

  16. Bioadhesive control of plasma proteins and blood cells from umbilical cord blood onto the interface grafted with zwitterionic polymer brushes.

    PubMed

    Chang, Yu; Chang, Yung; Higuchi, Akon; Shih, Yu-Ju; Li, Pei-Tsz; Chen, Wen-Yih; Tsai, Eing-Mei; Hsiue, Ging-Ho

    2012-03-06

    In this work, bioadhesive behavior of plasma proteins and blood cells from umbilical cord blood (UCB) onto zwitterionic poly(sulfobetaine methacrylate) (polySBMA) polymer brushes was studied. The surface coverage of polySBMA brushes on a hydrophobic polystyrene (PS) well plate with surface grafting weights ranging from 0.02 mg/cm(2) to 0.69 mg/cm(2) can be effectively controlled using the ozone pretreatment and thermal-induced radical graft-polymerization. The chemical composition, grafting structure, surface hydrophilicity, and hydration capability of prepared polySBMA brushes were determined to illustrate the correlations between grafting properties and blood compatibility of zwitterionic-grafted surfaces in contact with human UCB. The protein adsorption of fibrinogen in single-protein solutions and at complex medium of 100% UCB plasma onto different polySBMA brushes with different grafting coverage was measured by enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies. The grafting density of the zwitterionic brushes greatly affects the PS surface, thus controlling the adsorption of fibrinogen, the adhesion of platelets, and the preservation of hematopoietic stem and progenitor cells (HSPCs) in UCB. The results showed that PS surfaces grafted with polySBMA brushes possess controllable hydration properties through the binding of water molecules, regulating the bioadhesive and bioinert characteristics of plasma proteins and blood platelets in UCB. Interestingly, it was found that the polySBMA brushes with an optimized grafting weight of approximately 0.1 mg/cm(2) at physiologic temperatures show significant hydrated chain flexibility and balanced hydrophilicity to provide the best preservation capacity for HSPCs stored in 100% UCB solution for 2 weeks. This work suggests that, through controlling grafting structures, the hemocompatible nature of grafted zwitterionic polymer brushes makes them well suited to the molecular design of regulated

  17. Rhodopsin photoactivation dynamics revealed by quasi-elastic neutron scattering

    DOE PAGES

    Bhowmik, Debsindhu; Shrestha, Utsab; Perera, Suchithranga M.d.c.; ...

    2015-01-27

    Rhodopsin is a G-protein-coupled receptor (GPCR) responsible for vision under dim light conditions. During rhodopsin photoactivation, the chromophore retinal undergoes cis-trans isomerization, and subsequently dissociates from the protein yielding the opsin apoprotein [1]. What are the changes in protein dynamics that occur during the rhodopsin photoactivation process? Here, we studied the microscopic dynamics of the dark-state rhodopsin and the ligand-free opsin using quasi-elastic neutron scattering (QENS). The QENS technique tracks the individual hydrogen atom motions in the protein molecules, because the neutron scattering cross-section of hydrogen is much higher than other atoms [2-4]. We used protein (rhodopsin/opsin) samples with CHAPSmore » detergent hydrated with heavy water. The solvent signal is suppressed due to the heavy water, so that only the signals from proteins and detergents are detected. The activation of proteins is confirmed at low temperatures up to 300 K by the mean-square displacement (MSD) analysis. Our QENS experiments conducted at temperatures ranging from 220 K to 300 K clearly indicate that the protein dynamic behavior increases with temperature. The relaxation time for the ligand-bound protein rhodopsin was longer compared to opsin, which can be correlated with the photoactivation. Moreover, the protein dynamics are orders of magnitude slower than the accompanying CHAPS detergent, which forms a band around the protein molecule in the micelle. Unlike the protein, the CHAPS detergent manifests localized motions that are the same as in the bulk empty micelles. Furthermore QENS provides unique understanding of the key dynamics involved in the activation of the GPCR involved in the visual process.« less

  18. Perturbations of Native Membrane Protein Structure in Alkyl Phosphocholine Detergents: A Critical Assessment of NMR and Biophysical Studies

    PubMed Central

    2018-01-01

    Membrane proteins perform a host of vital cellular functions. Deciphering the molecular mechanisms whereby they fulfill these functions requires detailed biophysical and structural investigations. Detergents have proven pivotal to extract the protein from its native surroundings. Yet, they provide a milieu that departs significantly from that of the biological membrane, to the extent that the structure, the dynamics, and the interactions of membrane proteins in detergents may considerably vary, as compared to the native environment. Understanding the impact of detergents on membrane proteins is, therefore, crucial to assess the biological relevance of results obtained in detergents. Here, we review the strengths and weaknesses of alkyl phosphocholines (or foscholines), the most widely used detergent in solution-NMR studies of membrane proteins. While this class of detergents is often successful for membrane protein solubilization, a growing list of examples points to destabilizing and denaturing properties, in particular for α-helical membrane proteins. Our comprehensive analysis stresses the importance of stringent controls when working with this class of detergents and when analyzing the structure and dynamics of membrane proteins in alkyl phosphocholine detergents. PMID:29488756

  19. Phospholipase A2 activity-dependent and -independent fusogenic activity of Naja nigricollis CMS-9 on zwitterionic and anionic phospholipid vesicles.

    PubMed

    Chiou, Yi-Ling; Chen, Ying-Jung; Lin, Shinne-Ren; Chang, Long-Sen

    2011-11-01

    CMS-9, a phospholipase A(2) (PLA(2)) from Naja nigricollis venom, induced the death of human breast cancer MCF-7 cells accompanied with the formation of cell clumps without clear boundaries between cells. Annexin V-FITC staining indicated that abundant phosphatidylserine appeared on the outer membrane of MCF-7 cell clumps, implying the possibility that CMS-9 may promote membrane fusion via anionic phospholipids. To validate this proposition, fusogenic activity of CMS-9 on vesicles composed of zwitterionic phospholipid alone or a combination of zwitterionic and anionic phospholipids was examined. Although CMS-9-induced fusion of zwitterionic phospholipid vesicles depended on PLA(2) activity, CMS-9-induced fusion of vesicles containing anionic phospholipids could occur without the involvement of PLA(2) activity. Membrane-damaging activity of CMS-9 was associated with its fusogenicity. Moreover, CMS-9 induced differently membrane leakage and membrane fusion of vesicles with different compositions. Membrane fluidity and binding capability with phospholipid vesicles were not related to the fusogenicity of CMS-9. However, membrane-bound conformation and mode of CMS-9 depended on phospholipid compositions. Collectively, our data suggest that PLA(2) activity-dependent and -independent fusogenicity of CMS-9 are closely related to its membrane-bound modes and targeted membrane compositions. Copyright © 2011 Elsevier Ltd. All rights reserved.

  20. Pigmented and hyperkeratotic napkin dermatitis: a liquid detergent irritant dermatitis.

    PubMed

    Patrizi, A; Neri, I; Marzaduri, S; Fiorillo, L

    1996-01-01

    Napkin or diaper dermatitis (DD) is an inflammatory cutaneous eruption limited to the diaper area and common in the first 2 years of life. A number of clinical variants of DD have been identified. We report a new variant of DD characterized by papyraceous skin, brownish discoloration and predilection for the depth of folds. 15 infants and toddlers affected by this peculiar type of DD were evaluated regarding duration, localization, morphology and evolution of their dermatosis. This variant of DD was mainly confined to the depth of inguinal and gluteal folds and invariably associated with severe xerosis with papyraceous and glazed skin. The patients were healthy and asymptomatic and all laboratory investigations performed were normal. All patients were frequently changed and thoroughly washed with synthetic detergents with acid pH. DD improved rapidly with reduced frequency of washing and discontinuation of liquid detergents. We conclude that this condition is a type of irritant contact dermatitis from excessive use of lipid acid detergents.

  1. Development and reduction of hypertension and oxidative stress among detergent industry workers.

    PubMed

    Boojar, Massod M A; Goodarzi, Faranak; Boojar, Manochehr M A

    2004-12-01

    Hypertension status and oxidative stress parameters were assessed in 291 workers (hypertensive workers were divided into three grades, non-equivalently) at two detergent production plants, one of which included enzymes in the detergent (n=138) and another which did not (n=153), and 45 control workers in another industry three times (at the time of employment, 7 yrs later at the time of installation of a filter system, and about 3 yrs later). Malondialdehyde (MDA) was measured by high-performance liquid chromatography, antioxidant enzymes and lipid status by ultraviolet-visible spectrophotometry, trace elements by atomic absorption spectroscopy, and blood pressure using an oscilometric device. Prior to filter system installation, enzyme-exposed workers had significantly higher MDA, antioxidant enzyme activities, and prevalence of hypertension, compared with controls. The filter system reduced airborne detergent and enzyme dusts, resulting in a decreased prevalence of hypertension and a significant improvement in workers' oxidative stress indicators. Alterations in antioxidant status may result from the cumulative effect of high levels of detergent and enzyme in airborne dust in the workplace.

  2. The Effect Of Additional Detergent In Crude Palm Oil In The Process Of Separation Stearin

    NASA Astrophysics Data System (ADS)

    Rezekyah Hasibuan, Vina; aini, Nur; Febriyanti; Ayubi Pane, Salahudin Al

    2018-03-01

    This study aims to find out how much stearin is formed from the addition of detergent and to understand the process of separation of crude olein with crude stearin from raw material of crude palm oil (CPO). Using a detergent fractionation system, detergent fractionation is a continuous crystallization of oil with controlled cooling and the separation of fractions by weight or centrifuge after supplementing surfactant.

  3. Waste prevention in liquid detergent distribution: a comparison based on life cycle assessment.

    PubMed

    Nessi, Simone; Rigamonti, Lucia; Grosso, Mario

    2014-11-15

    The distribution of liquid detergents through self-dispensing systems has been adopted in some Italian retail stores over the last few years. By enabling the consumer to refill several times the same container, it is proposed as a less waste-generating and more environmentally friendly alternative to the traditional distribution with single-use plastic containers. For this reason, its implementation is encouraged by the national waste prevention programme recently adopted in Italy. In order to assess such claims, a life cycle assessment was carried out to evaluate whether detergent distribution through self-dispensing systems actually allows to achieve the expected reduction in waste generation and environmental impacts. The focus was on the distribution within the large-scale retail trade and on the categories of laundry detergents, fabric softeners and hand dishwashing detergents. For each of them, a set of baseline single-use scenarios were compared with two alternative waste prevention scenarios, where the detergent is distributed through self-dispensing systems. Beyond waste generation, also the Cumulative Energy Demand and thirteen midpoint-level potential impact indicators were calculated for the comparison. Results showed that a reduction in waste generation up to 98% can be achieved, depending on the category of detergent, on the baseline scenario of comparison and on the number of times the refillable container is used. A progressive reduction in the energy demand and in most of the potential impacts was also observed, starting from a minimum number of uses of the refillable container. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Physical-chemical features of non-detergent sulfobetaines active as protein-folding helpers.

    PubMed

    Expert-Bezançon, Nicole; Rabilloud, Thierry; Vuillard, Laurent; Goldberg, Michel E

    2003-01-01

    Some non-detergent sulfobetaines had been shown to prevent aggregation and improve the yield of active proteins when added to the buffer during in vitro protein renaturation. With the aim of designing more efficient folding helpers, a series of non-detergent sulfobetaines have been synthesized and their efficiency in improving the renaturation of a variety of proteins (E. coli tryptophan synthase and beta-D-galactosidase, hen lysozyme, bovine serum albumin, a monoclonal antibody) have been investigated. Attempts to correlate the structure of each sulfobetaines with its effect on folding revealed some molecular features that appear important in helping renaturation. This enabled us to design and synthesize new non-detergent sulfobetaines that act as potent folding helpers.

  5. Cooperative Effects of Zwitterionic-Ionic Surfactant Mixtures on the Interfacial Water Structure Revealed by Sum Frequency Generation Vibrational Spectroscopy.

    PubMed

    Pan, Xuecong; Yang, Fangyuan; Chen, Shunli; Zhu, Xuefeng; Wang, Chuanyi

    2018-05-08

    Cooperative effects of a series of equimolar binary zwitterionic-ionic surfactant mixtures on the interfacial water structure at the air-water interfaces have been studied by sum frequency generation vibrational spectroscopy (SFG-VS). For zwitterionic surfactant palmityl sulfobetaine (SNC 16 ), anionic surfactant sodium hexadecyl sulfate (SHS), and cationic surfactant cetyltrimethylammonium bromide (CTAB) with the same length of alkyl chain, significantly enhanced ordering of interfacial water molecules was observed for the zwitterionic-anionic surfactant mixtures SNC 16 -SHS, indicating that SNC 16 interacts more strongly with SHS than with CTAB because of the strong headgroup-headgroup electrostatic attraction for SNC 16 -SHS. Meanwhile, the SFG amplitude ratio of methyl and methylene symmetric stretching modes was used to verify the stronger interaction between SNC 16 and SHS. The conformational order indicator increased from 0.64 for SNC 16 to 7.17 for SNC 16 -SHS but only 0.94 for SNC 16 -CTAB. In addition, another anionic surfactant sodium dodecyl sulfate (SDS) was introduced to study the influence of chain-chain interaction. Decreased SFG amplitude of interfacial water molecules for SNC 16 -SDS was observed. Therefore, both the headgroup-headgroup electrostatic interaction and chain-chain van der Waals attractive interaction of the surfactants play an important role in enhancing the ordering of interfacial water molecules. The results provided experimental and theoretical bases for practical applications of the surfactants.

  6. A zwitterionic squaraine dye with a large Stokes shift for in vivo and site-selective protein sensing.

    PubMed

    Xu, Yongqian; Liu, Qin; Li, Xiaopeng; Wesdemiotis, Chrys; Pang, Yi

    2012-11-28

    A novel squaraine dye (SQ) exhibits improved fluorescence response toward protein detection by incorporation of a zwitterionic structure. With the aid of a dansylamide (DNSA) substituent, the new probe (DNSA-SQ) exhibits remarkable selectivity in binding to site I (a specific substructure in protein).

  7. Laundry Detergency of Solid Non-Particulate Soil Using Microemulsion-Based Formulation.

    PubMed

    Chanwattanakit, Jarussri; Chavadej, Sumaeth

    2018-02-01

    Laundry detergency of solid non-particulate soil on polyester and cotton was investigated using a microemulsion-based formulation, consisting of an anionic extended surfactant (C 12,13 -4PO-SO 4 Na) and sodium mono-and di-methyl naphthalene sulfonate (SMDNS) as the hydrophilic linker, to provide a Winsor Type III microemulsion with an ultralow interfacial tension (IFT). In this work, methyl palmitate (palmitic acid methyl ester) having a melting point around 30°C, was used as a model solid non-particulate (waxy) soil. A total surfactant concentration of 0.35 wt% of the selected formulation (4:0.65 weight ratio of C 12,13 -4PO-SO 4 Na:SMDNS) with 5.3 wt% NaCl was able to form a middle phase microemulsion at a high temperature (40°C),which provided the highest oil removal level with the lowest oil redeposition and the lowest IFT, and was much higher than that with a commercial detergent or de-ionized water. Most of the detached oil, whether in liquid or solid state, was in an unsolubilized form. Hence, the dispersion stability of the detached oil droplets or solidified oil particles that resulted from the surfactant adsorption played an important role in the oil redeposition. For an oily detergency, the lower the system IFT, the higher the oil removal whereas for a waxy (non-particulate) soil detergency, the lower the contact angle, the higher the solidified oil removal. For a liquefied oil, the detergency mechanism was roll up and emulsification with dispersion stability, while that for the waxy soil (solid oil) was the detachment by wettability with dispersion stability.

  8. Foam Properties and Detergent Abilities of the Saponins from Camellia oleifera

    PubMed Central

    Chen, Yu-Fen; Yang, Chao-Hsun; Chang, Ming-Shiang; Ciou, Yong-Ping; Huang, Yu-Chun

    2010-01-01

    The defatted seed meal of Camellia oleifera has been used as a natural detergent and its extract is commercially utilized as a foam-stabilizing and emulsifying agent. The goal of this study was to investigate the foam properties and detergent ability of the saponins from the defatted seed meal of C. oleifera. The crude saponin content in the defatted seed meal of C. oleifera was 8.34 and the total saponins content in the crude saponins extract was 39.5% (w/w). The foaming power of the 0.5 crude saponins extract solution from defatted seed meal of C. oleifera was 37.1 of 0.5 SLS solution and 51.3% to that of 0.5% Tween 80 solution. The R5 value of 86.0% represents good foam stability of the crude saponins extracted from the defatted seed meal of the plant. With the reduction of water surface tension from 72 mN/m to 50.0 mN/m, the 0.5% crude saponins extract solution has wetting ability. The sebum-removal experiment indicated that the crude saponins extract has moderate detergency. The detergent abilities of the saponins from C. oleifera and Sapindus mukorossi were also compared. PMID:21151446

  9. Jatropha oil derived sophorolipids: production and characterization as laundry detergent additive.

    PubMed

    Joshi-Navare, Kasturi; Khanvilkar, Poonam; Prabhune, Asmita

    2013-01-01

    Sophorolipids (SLs) are glycolipidic biosurfactants suitable for various biological and physicochemical applications. The nonedible Jatropha oil has been checked as the alternative raw material for SL synthesis using C. bombicola (ATCC22214). This is useful towards lowering the SL production cost. Through optimization of fermentation parameters and use of resting cell method, the yield 15.25 g/L could be achieved for Jatropha oil derived SL (SLJO) with 1% v/v oil feeding. The synthesized SL displayed good surfactant property. It reduced the surface tension of distilled water from 70.7 mN/m to 33.5 mN/m with the Critical Micelle Concentration (CMC) value of 9.5 mg/L. Keeping the prospective use of the SL in mind, the physicochemical properties were checked along with emulsion stability under temperature, pH stress, and in hard water. Also antibacterial action and stain removal capability in comparison with commercial detergent was demonstrated. SLJO enhanced the detergent performance. Based on the results, it can be said that SLs have utility as fabric cleaner with advantageous properties such as skin friendly nature, antibacterial action, and biodegradability. Therefore SLs are potential green molecules to replace synthetic surfactants in detergents so as to reduce harm caused to environment through detergent usage.

  10. Designer lipid-like peptides: a class of detergents for studying functional olfactory receptors using commercial cell-free systems.

    PubMed

    Corin, Karolina; Baaske, Philipp; Ravel, Deepali B; Song, Junyao; Brown, Emily; Wang, Xiaoqiang; Wienken, Christoph J; Jerabek-Willemsen, Moran; Duhr, Stefan; Luo, Yuan; Braun, Dieter; Zhang, Shuguang

    2011-01-01

    A crucial bottleneck in membrane protein studies, particularly G-protein coupled receptors, is the notorious difficulty of finding an optimal detergent that can solubilize them and maintain their stability and function. Here we report rapid production of 12 unique mammalian olfactory receptors using short designer lipid-like peptides as detergents. The peptides were able to solubilize and stabilize each receptor. Circular dichroism showed that the purified olfactory receptors had alpha-helical secondary structures. Microscale thermophoresis suggested that the receptors were functional and bound their odorants. Blot intensity measurements indicated that milligram quantities of each olfactory receptor could be produced with at least one peptide detergent. The peptide detergents' capability was comparable to that of the detergent Brij-35. The ability of 10 peptide detergents to functionally solubilize 12 olfactory receptors demonstrates their usefulness as a new class of detergents for olfactory receptors, and possibly other G-protein coupled receptors and membrane proteins.

  11. Designer Lipid-Like Peptides: A Class of Detergents for Studying Functional Olfactory Receptors Using Commercial Cell-Free Systems

    PubMed Central

    Corin, Karolina; Baaske, Philipp; Ravel, Deepali B.; Song, Junyao; Brown, Emily; Wang, Xiaoqiang; Wienken, Christoph J.; Jerabek-Willemsen, Moran; Duhr, Stefan; Luo, Yuan; Braun, Dieter; Zhang, Shuguang

    2011-01-01

    A crucial bottleneck in membrane protein studies, particularly G-protein coupled receptors, is the notorious difficulty of finding an optimal detergent that can solubilize them and maintain their stability and function. Here we report rapid production of 12 unique mammalian olfactory receptors using short designer lipid-like peptides as detergents. The peptides were able to solubilize and stabilize each receptor. Circular dichroism showed that the purified olfactory receptors had alpha-helical secondary structures. Microscale thermophoresis suggested that the receptors were functional and bound their odorants. Blot intensity measurements indicated that milligram quantities of each olfactory receptor could be produced with at least one peptide detergent. The peptide detergents' capability was comparable to that of the detergent Brij-35. The ability of 10 peptide detergents to functionally solubilize 12 olfactory receptors demonstrates their usefulness as a new class of detergents for olfactory receptors, and possibly other G-protein coupled receptors and membrane proteins. PMID:22132066

  12. A new hydrogen peroxide--based medical-device detergent with germicidal properties: comparison with enzymatic cleaners.

    PubMed

    Alfa, M J; Jackson, M

    2001-06-01

    The objective of this study was to evaluate the efficacy of the cleaning and bacterial killing ability of a new non-enzyme-based formulation (killing detergent solution [KDS]) compared with commercially available enzymatic detergents that included Metrizyme (Metrex Research Division of Sybron Canada Ltd. Morrisburg, Ontario) and Gzyme (Germiphene Corp, Brantford, Ontario). KDS is a hydrogen peroxide-based detergent formulation that combines cleaning efficacy with the ability to kill microorganisms. The KDS formulation helps ensure the protection of the health care worker from infectious risk during the soaking and cleaning stages of medical device reprocessing and reduces the bioburden on devices before sterilization/disinfection. Test organisms that included Enterococcus faecalis, Salmonella choleraesuis, Staphylococcus aureus, and Pseudomonas aeruginosa were suspended in artificial test soil (ATS-B; patent submitted), inoculated at 10(6) colonyforming units per carrier and dried overnight before detergent exposure. The ATS-B mimics the blood, protein, carbohydrate, and endotoxin levels of patient-used medical devices. Plastic lumen carriers and a flexible colonoscope were used for surface and simulated-use testing, respectively. The results for the microbial challenge dried onto polyvinyl chloride (PVC) carriers demonstrated that the ability of KDS to remove protein, blood, carbohydrate, and endotoxin from surface test carriers was as effective as the enzyme detergents that were evaluated. Furthermore, KDS was able to effect approximately a 5-Log(10) reduction in microbial loads with a 3-minute exposure at room temperature, whereas none of the other detergents were as effective. In simulated-use testing of a soiled colonoscope, KDS was significantly better at ensuring microbial killing compared with Gzyme and Metrizyme and was equivalent to the enzymatic detergents in cleaning ability. In summary the KDS has excellent microbial-killing ability in 3-minute

  13. Action on ileal smooth muscle of synthetic detergents and pardaxin.

    PubMed

    Primor, N

    1986-01-01

    Pardaxin (PX), a toxic and repellent substance isolated from the Red Sea flatfish, causes a sharp ball-like profile of drop of saline placed on a hydrophobic film to turn into a flattened one. This effect results with a decrease of the contact angle (theta) from 96 degrees to a maximum of 42 degrees at 10(-4) M of PX. The action of sodium dodecyl sulphate (SDS), a synthetic anionic detergent, benzalkonium chloride (BAC) cationic detergent and pardaxin (PX) a toxic protein with detergent properties, were studied in the ileal guinea-pig longitudinal smooth muscle preparation. SDS (4 X 10(-4) M) and PX (5 X 10(-6) M) diminished the muscle contractile response to field stimulation (0.1 Hz, 1 msec) and to acetylcholine (Ach) and to histamine and elicited a prolonged (4-6 min) TTX-insensitive muscle contraction. The dose dependence of muscle contraction to SDS and PX was found to be sigmoidal and occurred over a narrow range of concentrations. The SDS- but not PX-induced muscle contraction could be reduced by diphenhydramine (H1 antihistamine). BAC (10(-5)-10(-4) M) suppressed the muscle's contractile response to electrical stimulation (0.1 Hz, 1 msec), to Ach, histamine and 5-hydroxytryptamine but did not produce muscle contraction. PX at concentrations higher than 5 X 10(-6) M is a potent detergent and at this concentration shares several pharmacological similarities with SDS.

  14. Identification and absolute quantification of enzymes in laundry detergents by liquid chromatography tandem mass spectrometry.

    PubMed

    Gaubert, Alexandra; Jeudy, Jérémy; Rougemont, Blandine; Bordes, Claire; Lemoine, Jérôme; Casabianca, Hervé; Salvador, Arnaud

    2016-07-01

    In a stricter legislative context, greener detergent formulations are developed. In this way, synthetic surfactants are frequently replaced by bio-sourced surfactants and/or used at lower concentrations in combination with enzymes. In this paper, a LC-MS/MS method was developed for the identification and quantification of enzymes in laundry detergents. Prior to the LC-MS/MS analyses, a specific sample preparation protocol was developed due to matrix complexity (high surfactant percentages). Then for each enzyme family mainly used in detergent formulations (protease, amylase, cellulase, and lipase), specific peptides were identified on a high resolution platform. A LC-MS/MS method was then developed in selected reaction monitoring (SRM) MS mode for the light and corresponding heavy peptides. The method was linear on the peptide concentration ranges 25-1000 ng/mL for protease, lipase, and cellulase; 50-1000 ng/mL for amylase; and 5-1000 ng/mL for cellulase in both water and laundry detergent matrices. The application of the developed analytical strategy to real commercial laundry detergents enabled enzyme identification and absolute quantification. For the first time, identification and absolute quantification of enzymes in laundry detergent was realized by LC-MS/MS in a single run. Graphical Abstract Identification and quantification of enzymes by LC-MS/MS.

  15. Effects of Detergents on Catalytic Activity of Human Endometase/Matrilysin-2, a Putative Cancer Biomarker†

    PubMed Central

    Park, Hyun I.; Lee, Seakwoo; Ullah, Asad; Cao, Qiang; Sang, Qing-Xiang Amy

    2009-01-01

    Matrix metalloproteinases (MMPs) are a family of hydrolytic enzymes that play significant roles in development, morphogenesis, inflammation, and cancer invasion. Endometase (matrilysin 2 or MMP-26) is a putative early biomarker for human carcinomas. The effects of the ionic and nonionic detergents on catalytic activity of endometase were investigated. The hydrolytic activity of endometase was detergent concentration-dependent exhibiting a bell-shaped curve with its maximum activity near the critical micelle concentration (CMC) of nonionic detergents tested. The effect of Brij-35 on human gelatinase B (MMP-9), matirilysin (MMP-7), and membrane-type 1 MMP (MT1-MMP) was further explored. Their maximum catalysis was observed near the CMC of Brij-35 (~90 μM). Their IC50 values were above the CMC. The inhibition mechanism of MMP-7, MMP-9, and MT1-MMP by Brij-35 was mixed-type as determined by Dixon’s plot, however, that of endometase was non-competitive with a Ki value of 240 μM. The catalytic activities of MMPs are influenced by detergents. Monomer of detergents may activate and stabilize MMPs to enhance catalysis, but micelle of detergents may sequester enzyme and block substrate binding site to impede catalysis. Under physiological conditions lipid or membrane microenvironment may regulate enzymatic activity. PMID:19818727

  16. Insolubility and redistribution of GPI-anchored proteins at the cell surface after detergent treatment.

    PubMed Central

    Mayor, S; Maxfield, F R

    1995-01-01

    A diverse set of cell surface eukaryotic proteins including receptors, enzymes, and adhesion molecules have a glycosylphosphoinositol-lipid (GPI) modification at the carboxy-terminal end that serves as their sole means of membrane anchoring. These GPI-anchored proteins are poorly solubilized in nonionic detergent such as Triton X-100. In addition these detergent-insoluble complexes from plasma membranes are significantly enriched in several cytoplasmic proteins including nonreceptor-type tyrosine kinases and caveolin/VIP-21, a component of the striated coat of caveolae. These observations have suggested that the detergent-insoluble complexes represent purified caveolar membrane preparations. However, we have recently shown by immunofluorescence and electron microscopy that GPI-anchored proteins are diffusely distributed at the cell surface but may be enriched in caveolae only after cross-linking. Although caveolae occupy only a small fraction of the cell surface (< 4%), almost all of the GPI-anchored protein at the cell surface becomes incorporated into detergent-insoluble low-density complexes. In this paper we show that upon detergent treatment the GPI-anchored proteins are redistributed into a significantly more clustered distribution in the remaining membranous structures. These results show that GPI-anchored proteins are intrinsically detergent-insoluble in the milieu of the plasma membrane, and their co-purification with caveolin is not reflective of their native distribution. These results also indicate that the association of caveolae, GPI-anchored proteins, and signalling proteins must be critically re-examined. Images PMID:7579703

  17. Disinfection of the Skin with Detergent Preparations of Irgasan DP 300 and Other Antiseptics

    PubMed Central

    Lilly, H. A.; Lowbury, E. J. L.

    1974-01-01

    An evaluation of the relative effectiveness of 2% hexachlorophane and 0·75% Irgasan DP 300 bar soaps in disinfection of the hands showed that the former caused a significantly larger reduction in natural skin bacteria than the latter after one handwash and after six handwashings, three on each of two successive days. Repeated use of Irgasan DP 300 bar soap caused a significantly greater reduction in skin flora than repeated handwashings with unmedicated bar soap, but a single handwash gave no significant reduction in skin flora compared with a single use of the unmedicated soap. In a comparison of a 4% chlorhexidine detergent solution a 3% hexachlorophane detergent cream and a 2% Irgasan DP 300 detergent solution the 4% chlorhexidine detergent gave the largest mean reduction in skin bacteria after one handwash and after six handwashings and 2% Irgasan DP 300 a poor and erratic reduction after a single handwash. After six handwashings all three preparations gave large reductions in skin bacteria. The 2% Irgasan preparation showed some residual activity on the skin after handwashing though less than that with chlorhexidine and with hexachlorophane-chlorocresol detergent preparations. PMID:4609556

  18. Molecular assessment of collagen denaturation in decellularized tissues using a collagen hybridizing peptide.

    PubMed

    Hwang, Jeongmin; San, Boi Hoa; Turner, Neill J; White, Lisa J; Faulk, Denver M; Badylak, Stephen F; Li, Yang; Yu, S Michael

    2017-04-15

    Decellularized extracellular matrix (ECM) derived from tissues and organs are emerging as important scaffold materials for regenerative medicine. Many believe that preservation of the native ECM structure during decellularization is highly desirable. However, because effective techniques to assess the structural damage in ECM are lacking, the disruptive effects of a decellularization method and the impact of the associated structural damage upon the scaffold's regenerative capacity are often debated. Using a novel collagen hybridizing peptide (CHP) that specifically binds to unfolded collagen chains, we investigated the molecular denaturation of collagen in the ECM decellularized by four commonly used cell-removing detergents: sodium dodecyl sulfate (SDS), 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), sodium deoxycholate (SD), and Triton X-100. Staining of the detergent-treated porcine ligament and urinary bladder matrix with carboxyfluorescein-labeled CHP demonstrated that SDS and Triton X-100 denature the triple helical collagen molecule while CHAPS and SD do not, although second harmonic generation imaging and transmission electron microscopy (TEM) revealed that all four detergents disrupt collagen fibrils. Our findings from the CHP staining were further confirmed by the circular dichroism spectra of intact triple helical collagen molecules in CHAPS and SD solutions, and the TEM images of CHP-conjugated gold nanoparticles binding only to the SDS and Triton X-100 treated collagen fibrils. CHP is a powerful new tool for direct and reliable measurement of denatured collagen molecules in decellularized tissues. It is expected to have wide applications in the development and standardization of the tissue/organ decellularization technology. Preservation of the native ECM structure in decellularized tissues is highly desirable, since denaturation of ECM molecules (e.g., collagen) during decellularization can strongly influence the cellular response

  19. Molecular assessment of collagen denaturation in decellularized tissues using a collagen hybridizing peptide

    PubMed Central

    Hwang, Jeongmin; San, Boi Hoa; Turner, Neill J.; White, Lisa J.; Faulk, Denver M.; Badylak, Stephen F.; Li, Yang; Yu, S. Michael

    2017-01-01

    Decellularized extracellular matrix (ECM) derived from tissues and organs are emerging as important scaffold materials for regenerative medicine. Many believe that preservation of the native ECM structure during decellularization is highly desirable. However, because effective techniques to assess the structural damage in ECM are lacking, the disruptive effects of a decellularization method and the impact of the associated structural damage upon the scaffold’s regenerative capacity are often debated. Using a novel collagen hybridizing peptide (CHP) that specifically binds to unfolded collagen chains, we investigated the molecular denaturation of collagen in the ECM decellularized by four commonly used cellremoving detergents: sodium dodecyl sulfate (SDS), 3-[(3-cholamidopropyl)dimethylammonio]-1-propa nesulfonate (CHAPS), sodium deoxycholate (SD), and Triton X-100. Staining of the detergent-treated porcine ligament and urinary bladder matrix with carboxyfluorescein-labeled CHP demonstrated that SDS and Triton X-100 denature the triple helical collagen molecule while CHAPS and SD do not, although second harmonic generation imaging and transmission electron microscopy (TEM) revealed that all four detergents disrupt collagen fibrils. Our findings from the CHP staining were further confirmed by the circular dichroism spectra of intact triple helical collagen molecules in CHAPS and SD solutions, and the TEM images of CHP-conjugated gold nanoparticles binding only to the SDS and Triton X-100 treated collagen fibrils. CHP is a powerful new tool for direct and reliable measurement of denatured collagen molecules in decellularized tissues. It is expected to have wide applications in the development and standardization of the tissue/organ decellularization technology. Statement of Significance Preservation of the native ECM structure in decellularized tissues is highly desirable, since denaturation of ECM molecules (e.g., collagen) during decellularization can strongly

  20. Influence of Zwitterions on Thermomechanical Properties and Morphology of Acrylic Copolymers: Implications for Electroactive Applications

    DTIC Science & Technology

    2011-09-30

    transducers from branched sulfonated polysulfones.7 The mechanical strength of the membranes drastically decreased upon introduction of ionic liquids to... liquids ,8 and zwitterionomers maintained mechanical strengths upon swelling with 10 wt % ionic liquid . Zwitterions are chargedmolecules that contain...water purification5 to biotechnology.6 A unique combination of physical properties of ionomeric membranes is the ionic con- ductivity of lowmolar mass

  1. Dispersing Zwitterions into Comb Polymers for Nonviral Transfection: Experiments and Molecular Simulation.

    PubMed

    Ghobadi, Ahmadreza F; Letteri, Rachel; Parelkar, Sangram S; Zhao, Yue; Chan-Seng, Delphine; Emrick, Todd; Jayaraman, Arthi

    2016-02-08

    Polymer-based gene delivery vehicles benefit from the presence of hydrophilic groups that mitigate the inherent toxicity of polycations and that provide tunable polymer-DNA binding strength and stable complexes (polyplexes). However, hydrophilic groups screen charge, and as such can reduce cell uptake and transfection efficiency. We report the effect of embedding zwitterionic sulfobetaine (SB) groups in cationic comb polymers, using a combination of experiments and molecular simulations. Ring-opening metathesis polymerization (ROMP) produced comb polymers with tetralysine (K4) and SB pendent groups. Dynamic light scattering, zeta potential measurements, and fluorescence-based experiments, together with coarse-grained molecular dynamics simulations, described the effect of SB groups on the size, shape, surface charge, composition, and DNA binding strength of polyplexes formed using these comb polymers. Experiments and simulations showed that increasing SB composition in the comb polymers decreased polymer-DNA binding strength, while simulations indicated that the SB groups distributed throughout the polyplex. This allows polyplexes to maintain a positive surface charge and provide high levels of gene expression in live cells. Notably, comb polymers with nearly 50 mol % SB form polyplexes that exhibit positive surface charge similarly as polyplexes formed from purely cationic comb polymers, indicating the ability to introduce an appreciable amount of SB functionality without screening surface charge. This integrated simulation-experimental study demonstrates the effectiveness of incorporating zwitterions in polyplexes, while guiding the design of new and effective gene delivery vectors.

  2. Probing the weak interaction of proteins with neutral and zwitterionic antifouling polymers.

    PubMed

    Wu, Jiang; Zhao, Chao; Hu, Rundong; Lin, Weifeng; Wang, Qiuming; Zhao, Jun; Bilinovich, Stephanie M; Leeper, Thomas C; Li, Lingyan; Cheung, Harry M; Chen, Shengfu; Zheng, Jie

    2014-02-01

    Protein-polymer interactions are of great interest in a wide range of scientific and technological applications. Neutral poly(ethylene glycol) (PEG) and zwitterionic poly(sulfobetaine methacrylate) (pSBMA) are two well-known nonfouling materials that exhibit strong surface resistance to proteins. However, it still remains unclear or unexplored how PEG and pSBMA interact with proteins in solution. In this work, we examine the interactions between two model proteins (bovine serum albumin and lysozyme) and two typical antifouling polymers of PEG and pSBMA in aqueous solution using fluorescence spectroscopy, atomic force microscopy and nuclear magnetic resonance. The effect of protein:polymer mass ratios on the interactions is also examined. Collective data clearly demonstrate the existence of weak hydrophobic interactions between PEG and proteins, while there are no detectable interactions between pSBMA and proteins. The elimination of protein interaction with pSBMA could be due to an enhanced surface hydration of zwitterionic groups in pSBMA. New evidence is given to demonstrate the interactions between PEG and proteins, which are often neglected in the literature because the PEG-protein interactions are weak and reversible, as well as the structural change caused by hydrophobic interaction. This work provides a better fundamental understanding of the intrinsic structure-activity relationship of polymers underlying polymer-protein interactions, which are important for designing new biomaterials for biosensor, medical diagnostics and drug delivery applications. Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  3. Evaluation of an ODS column modified with zwitterionic/nonionic mixed surfactants and its application to direct injection determination of inorganic anions.

    PubMed

    Hasegawa, Takuya; Umemura, Tomonari; Koide, Akira; Chiba, Koichi; Ueki, Yuji; Tsunoda, Kin-ichi; Haraguchi, Hiroki

    2005-08-01

    An octadecylsilica (ODS) column modified with zwitterionic/nonionic mixed surfactants was evaluated for the direct injection determination of inorganic anions in biological fluids by ion chromatography. A zwitterionic surfactant (sulfobetaine-type) and a nonionic surfactant (polyoxyethylene-type) were used for a stationary-phase modification. When aqueous electrolyte solutions with concentrations of sub-mM to several mM were used as a mobile phase, the zwitterionic surfactant coated on the ODS surface exhibited unique separation selectivity for ionic species, while the nonionic surfactant coated on the ODS might have formed a hydrophilic network over the ODS surface and restricted matrix proteins from adsorbing on the stationary phase. Consequently, the mixed surfactant-modified column system allowed an efficient ion chromatographic separation of inorganic anions as well as a size-exclusive removal of column-fouling proteins. This separation system was applied to the direct injection determination of UV-absorbing anions in human saliva. The detection limits for nitrite, nitrate, iodide and thiocyanate were 3.1, 2.7, 4.5 and 25 microM, respectively, with UV detection at 210 nm (injection volume; 20 microl), and their relative standard deviations for 5 replicate measurements of saliva samples spiked with 100 microM each of those anions were 1.4, 0.9, 2.2 and 5.5%, respectively.

  4. Tonoplast of Beta vulgaris L. contains detergent-resistant membrane microdomains.

    PubMed

    Ozolina, Natalia V; Nesterkina, Irina S; Kolesnikova, Ekaterina V; Salyaev, Ryurik K; Nurminsky, Vadim N; Rakevich, Alexander L; Martynovich, Evgueni F; Chernyshov, Michael Yu

    2013-03-01

    The experiments conducted on tonoplast of Beta vulgaris L. roots were performed to identify detergent-resistant lipid-protein microdomains (DRMs, interpreted as lipid rafts).The presence of DRMs can be found when dynamic clustering of sphingolipids, sterols, saturated fatty acids is registered, and the insolubility of these microdomains in nonionic detergents at low temperatures is proven. The elucidation of tonoplast microdomains has been based on results obtained with the aid of high-speed centrifuging in the sucrose gradient. The experiments have shown that tonoplast microdomains are rich in sphingolipids, free sterols and saturated fatty acids (such a lipid content is also typical of lipid-protein microdomains of other membranes), while only few phospholipids are present in tonoplast microdomains. The presence of microdomains has been confirmed by fluorescence and confocal microscopy using filipin and Laurdan as fluorescent probes. The experiments with Laurdan have shown that tonoplast microdomains are characterized by a high order compared to characteristics of the rest of the tonoplast. Thus, the presence of detergent-resistant lipid-protein microdomains in the tonoplast has been demonstrated.

  5. Ligand-induced association of surface immunoglobulin with the detergent insoluble cytoskeleton may involve an 89K protein

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

    Gupta, S.K.; Woda, B.

    1986-03-01

    Membrane immunoglobulin of B-lymphocytes is thought to play an important role in antigen recognition and cellular activation. Binding of cross-linking ligands to surface immunoglobulin (SIg) on intact cells converts it to a detergent insoluble state, and this conversion is associated with the transmission of a mitogenic signal. Insolubilized membrane proteins may be solubilized by incubating the detergent insoluble cytoskeletons in buffers which convert F-actin to G-actin ((Buffer 1), 0.34M sucrose, 0.5mM ATP, 0.5mM Dithiothrietol and lmM EDTA). Immunoprecipitation of SIg from the detergent soluble fraction of /sup 35/S-methionine labeled non ligand treated rat B-cells results in the co-isolation of anmore » 89K protein and a 44K protein, presumably actin. The 89K protein is not associated with the fraction of endogenous detergent insoluble SIg. On treatment of rat B cells with cross-linking ligand (anti-Ig) the 89K protein becomes detergent insoluble along with most of the SIg and co-isolates with SIg on immunoprecipitation of the detergent insoluble, buffer l solubilized fraction. The migration of the SIg-associated 89K protein from the detergent soluble fraction to the detergent insoluble fraction after ligand treatment, suggests that this protein might be involved in linking SIg to the underlying cytoskeleton and could be involved in the transmission of a mitogenic signal.« less

  6. Laundry detergent and possible nonaccidental injury.

    PubMed

    Howieson, Alan J; Harley, Oliver J H; Tiernan, Eunan P

    2007-06-01

    Nonaccidental injury is always a concern when children present with unusual injuries. The case of a child who presented with a partial thickness burn secondary to prolonged contact with a liquid biological laundry detergent is described. Initially there was some doubt as to whether the agent in question could cause this injury but a small experiment on a volunteer confirmed it was possible.

  7. Characterization of detergent compatible protease from halophilic Virgibacillus sp. CD6.

    PubMed

    Lam, Ming Quan; Nik Mut, Nik Nurhidayu; Thevarajoo, Suganthi; Chen, Sye Jinn; Selvaratnam, Chitra; Hussin, Huszalina; Jamaluddin, Haryati; Chong, Chun Shiong

    2018-02-01

    A halophilic bacterium, Virgibacillus sp. strain CD6, was isolated from salted fish and its extracellular protease was characterized. Protease production was found to be highest when yeast extract was used as nitrogen source for growth. The protease exhibited stability at wide range of salt concentration (0-12.5%, w/v), temperatures (20-60 °C), and pH (4-10) with maximum activity at 10.0% (w/v) NaCl, 60 °C, pH 7 and 10, indicating its polyextremophilicity. The protease activity was enhanced in the presence of Mg 2+ , Mn 2+ , Cd 2+ , and Al 3+ (107-122% relative activity), and with retention of activity > 80% for all of other metal ions examined (K + , Ca 2+ , Cu 2+ , Co 2+ , Ni 2+ , Zn 2+ , and Fe 3+ ). Both PMSF and EDTA inhibited protease activity, denoting serine protease and metalloprotease properties, respectively. High stability (> 70%) was demonstrated in the presence of organic solvents and detergent constituents, and the extracellular protease from strain CD6 was also found to be compatible in commercial detergents. Proteinaceous stain removal efficacy revealed that crude protease of strain CD6 could significantly enhance the performance of commercial detergent. The protease from Virgibacillus sp. strain CD6 could serve as a promising alternative for various applications, especially in detergent industry.

  8. Structure of water at zwitterionic copolymer film-liquid water interfaces as examined by the sum frequency generation method.

    PubMed

    Kondo, Takuya; Nomura, Kouji; Gemmei-Ide, Makoto; Kitano, Hiromi; Noguchi, Hidenori; Uosaki, Kohei; Saruwatari, Yoshiyuki

    2014-01-01

    A copolymer film composed of zwitterionic carboxymethylbetaine (CMB) and n-butyl methacrylate (BMA), Poly(CMB-r-BMA), was cast on a flat plane of an octadecyltrichlorosilane (ODS)-modified fused quartz prism with a semi-cylindrical shape. CH stretching of the polymer film and O-H stretching of water at the surface of the film were examined using the sum frequency generation (SFG) technique. The C-H stretching band of the cast film, indicating a gauche defect of the film, was affected by the contact medium including dry nitrogen, water vapor-saturated nitrogen and liquid water. In contrast, the C-H stretching of an octadecyl group introduced onto the quartz prism for stable attachment of the cast film was not significantly changed by the contact medium. The O-H stretching band indicated that water molecules at the surface of the Poly(CMB-r-BMA) film in contact with liquid water were not greatly oriented in comparison with those at the surfaces of a bare prism, an ODS SAM-modified prism, and a prism covered with a PolyBMA film or a copolymer film of BMA and methacrylic acid or 2-(dimethylamino)ethyl methacrylate. A similar small perturbation of the structure of water was previously observed in the vicinity of water-soluble zwitterionic polymers and zwitterionic copolymer films using Raman and attenuated total reflection infrared spectroscopies, respectively. A distinct effect of charge neutralization to diminish the perturbation of the structure of interfacial water around polymer materials was suggested. Copyright © 2013 Elsevier B.V. All rights reserved.

  9. 40 CFR 417.150 - Applicability; description of the manufacture of spray dried detergents subcategory.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 29 2011-07-01 2009-07-01 true Applicability; description of the manufacture of spray dried detergents subcategory. 417.150 Section 417.150 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) EFFLUENT GUIDELINES AND STANDARDS SOAP AND DETERGENT MANUFACTURING...

  10. 40 CFR 417.150 - Applicability; description of the manufacture of spray dried detergents subcategory.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 30 2012-07-01 2012-07-01 false Applicability; description of the manufacture of spray dried detergents subcategory. 417.150 Section 417.150 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) EFFLUENT GUIDELINES AND STANDARDS SOAP AND DETERGENT MANUFACTURING...

  11. 40 CFR 417.150 - Applicability; description of the manufacture of spray dried detergents subcategory.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 29 2014-07-01 2012-07-01 true Applicability; description of the manufacture of spray dried detergents subcategory. 417.150 Section 417.150 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) EFFLUENT GUIDELINES AND STANDARDS SOAP AND DETERGENT MANUFACTURING...

  12. Novel sulfonated polyimide/zwitterionic polymer-functionalized graphene oxide hybrid membranes for vanadium redox flow battery

    NASA Astrophysics Data System (ADS)

    Cao, Li; Kong, Lei; Kong, Lingqian; Zhang, Xingxiang; Shi, Haifeng

    2015-12-01

    Hybrid membranes (SPI/ZGO) composed of sulfonated polyimide (SPI) and zwitterionic polymer-functionalized graphene oxide (ZGO) are fabricated via a solution-casting method for vanadium redox flow battery (VRB). Successful preparation of ZGO fillers and SPI/ZGO hybrid membranes are demonstrated by FT-IR, XPS and SEM, indicating that ZGO fillers is homogeneously dispersed into SPI matrix. Through controlling the interfacial interaction between SPI matrix and ZGO fillers, the physicochemical properties, e.g., vanadium ion barrier and proton transport pathway, of hybrid membranes are tuned via the zwitterionic acid-base interaction in the hybrid membrane, showing a high ion selectivity and good stability with the incorporated ZGO fillers. SPI/ZGO-4 hybrid membrane proves a higher cell efficiencies (CE: 92-98%, EE: 65-79%) than commercial Nafion 117 membrane (CE: 89-94%, EE: 59-70%) for VRB application at 30-80 mA cm-2. The assembled VRB with SPI/ZGO-4 membrane presents a stable cycling charge-discharge performance over 280 times, which demonstrates its excellent chemical stability under the strong acidic and oxidizing conditions. SPI/ZGO hybrid membranes show a brilliant perspective for VRB application.

  13. PH-dependence of detergent-induced hemolysis and vesiculation of erythrocytes.

    PubMed

    Chernitsky, E A; Rozin, V V; Senkovich, O A

    2001-01-01

    The influence of pH of the medium on the parameters of detergent-induced fast hemolysis and vesiculation of human erythrocytes was studied. In the range of pH 6.3-7.2 neither the extent nor the rate of the vesiculation induced by 25 microM sodium dodecyl sulfate (SDS) changed. However, a decrease of pH from 8.0 to 5.8 strongly modified both the extent and the rate of the hemolysis induced by SDS. Within the range of pH 8.0-6.4, the effect can be ascribed to the increase of the positive charge of the membrane. This could lead to the accumulation of the membrane-bound anion detergent and, hence, to the change of the hemolysis parameters. Non-charged detergent Triton X-100 did not display any pH-dependence. At pH between 6.4 and 5.8 the extent and rate of hemolysis changed in a complicated manner. The kinetic curves of hemolysis could be approximated by a single exponential within the pH range between 8.0 and 7.2. Upon further reduction of pH, a second exponential component, with a larger time constant, appeared in the kinetic curves. At 5.8 < pH < 7.2, the contribution of the "fast" hemolysis dropped virtually to zero, with pK about 6.0. This points to a structural transition of the membrane, possibly involving histidine. We suggest that the parameters of the detergent-induced hemolysis are sensitive to the changes of the charge and structural state of erythrocyte membrane.

  14. Characterization of the activity and stability of amylase from saliva and detergent: laboratory practicals for studying the activity and stability of amylase from saliva and various commercial detergents.

    PubMed

    Valls, Cristina; Rojas, Cristina; Pujadas, Gerard; Garcia-Vallve, Santi; Mulero, Miquel

    2012-07-01

    This article presents two integrated laboratory exercises intended to show students the role of α-amylases (AAMYs) in saliva and detergents. These laboratory practicals are based on the determination of the enzymatic activity of amylase from saliva and different detergents using the Phadebas test (quantitative) and the Lugol test (qualitative) under different conditions (e.g. variations in temperature and alkalinity). This work also proposes the study of enzyme stability in the presence of several surfactants and oxidizing agents using the same technical approach. The proposed laboratory exercises promote the understanding of the physiological function of this enzyme and the biotechnological applications of AAMYs in the detergent industry. The exercises also promote the understanding that the enzymatic stability and performance are dependent on the organism of origin, and if necessary, these properties could be modified by genetic engineering. In addition, this article reinforces the development of laboratory skills, problem-solving capabilities, and the ability to write a laboratory report. The exercises are proposed primarily as an undergraduate project for advanced students in the biochemical and biotechnological sciences. These laboratory practicals are complementary to the previously published BAMBED article (Biochemistry and Molecular Biology Education Vol. 39, No. 4, pp. 280-290, 2011) on detergent proteases. Copyright © 2012 Wiley Periodicals, Inc.

  15. Anti-Fouling Double-Skinned Forward Osmosis Membrane with Zwitterionic Brush for Oily Wastewater Treatment.

    PubMed

    Ong, Chi Siang; Al-Anzi, Bader; Lau, Woei Jye; Goh, Pei Sean; Lai, Gwo Sung; Ismail, Ahmad Fauzi; Ong, Yue Seong

    2017-07-31

    Despite its attractive features for energy saving separation, the performance of forward osmosis (FO) has been restricted by internal concentration polarization and fast fouling propensity that occur in the membrane sublayer. These problems have significantly affected the membrane performance when treating highly contaminated oily wastewater. In this study, a novel double-skinned FO membrane with excellent anti-fouling properties has been developed for emulsified oil-water treatment. The double-skinned FO membrane comprises a fully porous sublayer sandwiched between a highly dense polyamide (PA) layer for salt rejection and a fairly loose dense bottom zwitterionic layer for emulsified oil particle removal. The top dense PA layer was synthesized via interfacial polymerization meanwhile the bottom layer was made up of a zwitterionic polyelectrolyte brush - (poly(3-(N-2-methacryloxyethyl-N,N-dimethyl) ammonatopropanesultone), abbreviated as PMAPS layer. The resultant double-skinned membrane exhibited a high water flux of 13.7 ± 0.3 L/m 2 .h and reverse salt transport of 1.6 ± 0.2 g/m 2 .h under FO mode using 2 M NaCl as the draw solution and emulsified oily solution as the feed. The double-skinned membrane outperforms the single-skinned membrane with much lower fouling propensity for emulsified oil-water separation.

  16. Electrophoretic separations in poly(dimethylsiloxane) microchips using a mixture of ionic and zwitterionic surfactants

    PubMed Central

    Guan, Qian; Noblitt, Scott D.; Henry, Charles S.

    2012-01-01

    The use of mixtures of ionic and zwitterionic surfactants in poly(dimethylsiloxane) (PDMS) microchips is reported. The effect of surfactant concentration on EOF was studied for a single anionic surfactant (sodium dodecyl sulfate, SDS), a single zwitterionic surfactant (N-tetradecylammonium-N,N-dimethyl-3-ammonio-1-propanesulfonate, TDAPS), and a mixed SDS/TDAPS surfactant system. SDS increased the EOF as reported previously while TDAPS showed an initial increase in EOF followed by a reduction at higher concentrations. When TDAPS was added to a solution containing SDS, the EOF decreased in a concentration dependent manner. The EOF for all three surfactant systems followed expected pH trends, with increasing EOF at higher pH. The mixed surfactant system allowed tuning of the EOF across a range of pH and concentration conditions. After establishing the EOF behavior, the adsorption/desorption kinetics were measured and showed a slower adsorption/desorption rate for TDAPS than SDS. Finally, the separation and electrochemical detection of model catecholamines in buffer and reduced glutathione (GSH) in red blood cell lysate using the mixed surfactant system were explored. The mixed surfactant system provided shorter analysis times and/or improved resolution when compared to the single surfactant systems. PMID:22222982

  17. Detergent Stabilized Nanopore Formation Kinetics of an Anthrax Protein

    NASA Astrophysics Data System (ADS)

    Peterson, Kelby

    2015-03-01

    This summer research project funded through the Society of Physics Students Internship Program and The National Institute of Standards and Technology focused on optimization of pore formation of Protective Antigen protein secreted by Bacillus Anthraces. This experiment analyzes the use of N-tetradecylphosphocholine (FOS-14 Detergent) to stabilize the water soluble protein, protective antigen protein (PA63) to regulate the kinetics of pore formation in a model bilayer lipid membrane. The FOS-14 Detergent was tested under various conditions to understand its impact on the protein pore formation. The optimization of this channel insertion is critical in preparing samples of oriented for neutron reflectometry that provide new data to increase the understanding of the protein's structure.

  18. Tetrakis(1-imidazolyl) borate (BIM4) based zwitterionic and related molecules used as electron injection layers

    DOEpatents

    Li, Huaping; Xu, Yunhua; Bazan, Guillermo C

    2013-02-05

    Tetrakis(1-imidazolyl)borate (BIm4) based zwitterionic and/or related molecules for the fabrication of PLEDs is provided. Device performances with these materials approaches that of devices with Ba/Al cathodes for which the cathode contact is ohmic. Methods of producing such materials, and electron injection layers and devices containing these materials are also provided.

  19. Efficacy of detergents and fresh produce disinfectants against microorganisms associated with mixed raw vegetables.

    PubMed

    Samadi, Nasrin; Abadian, Neda; Bakhtiari, Donya; Fazeli, Mohammad Reza; Jamalifar, Hossein

    2009-07-01

    Efficacy of commercial detergent and disinfectants to eliminate microorganisms associated with fresh vegetables eaten raw in Iran, including radish, parsley, basil, coriander (cilantro), Allium porrum (leek), and peppermint were studied. The raw vegetables were subjected to a triple wash treatment of washing in tap water for mud removal, washing in water containing a detergent (dishwashing liquid) or disinfectant individually, and rinsing in tap water. The population of total mesophilic microbes on the surface of untreated vegetables ranged from 10(5) to 10(6) CFU/g. Washing in tap water or treatment with detergent (333 ppm for 10 min) or benzalkonium chloride (92 ppm for 15 min) reduced the total microbial count, most probable number (MPN) of coliforms, MPN of fecal coliforms, and MPN of fecal streptococci by about 1.2 to 2.3 log. No significant differences in microbial populations were found on vegetables after decontamination with tap water, detergent, or benzalkonium chloride (P > 0.05). Treatments with peracetic acid (100 ppm for 15 min) and hydrogen peroxide (133 ppm for 30 min) reduced the total mesophilic microbial counts by about 2.8 log. The microbial reductions with calcium hypochlorite (300 ppm for 15 min) and combined hydrogen peroxide and silver ion (133 ppm for 30 min) were significantly higher than those obtained after rinsing in tap water or after detergent or benzalkonium chloride wash (P < 0.05). Pretreatment with detergent slightly enhanced the efficacy of all decontamination treatments, but results were not significantly different from those obtained after individual application of disinfectants.

  20. Protein unfolding in detergents: effect of micelle structure, ionic strength, pH, and temperature.

    PubMed Central

    Otzen, Daniel E

    2002-01-01

    The 101-residue monomeric protein S6 unfolds in the anionic detergent sodium dodecyl sulfate (SDS) above the critical micelle concentration, with unfolding rates varying according to two different modes. Our group has proposed that spherical micelles lead to saturation kinetics in unfolding (mode 1), while cylindrical micelles prevalent at higher SDS concentrations induce a power-law dependent increase in the unfolding rate (mode 2). Here I investigate in more detail how micellar properties affect protein unfolding. High NaCl concentrations, which induce cylindrical micelles, favor mode 2. This is consistent with our model, though other effects such as electrostatic screening cannot be discounted. Furthermore, unfolding does not occur in mode 2 in the cationic detergent LTAB, which is unable to form cylindrical micelles. A strong retardation of unfolding occurs at higher LTAB concentrations, possibly due to the formation of dead-end protein-detergent complexes. A similar, albeit much weaker, effect is seen in SDS in the absence of salt. Chymotrypsin inhibitor 2 exhibits the same modes of unfolding in SDS as S6, indicating that this type of protein unfolding is not specific for S6. The unfolding process in mode 1 has an activation barrier similar in magnitude to that in water, while the activation barrier in mode 2 is strongly concentration-dependent. The strong pH-dependence of unfolding in SDS and LTAB suggests that the rate of unfolding in anionic detergent is modulated by repulsion between detergent headgroups and anionic side chains, while cationic side chains modulate unfolding rates in cationic detergents. PMID:12324439

  1. Insight into the Structure of Light Harvesting Complex II and its Stabilization in Detergent Solution

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

    Cardoso, Mateus B; Smolensky, Dmitriy; Heller, William T

    2009-01-01

    The structure of spinach light-harvesting complex II (LHC II), stabilized in a solution of the detergent n-octyl-{beta}-d-glucoside (BOG), was investigated by small-angle neutron scattering (SANS). Physicochemical characterization of the isolated complex indicated that it was pure (>95%) and also in its native trimeric state. SANS with contrast variation was used to investigate the properties of the protein-detergent complex at three different H{sub 2}O/D{sub 2}O contrast match points, enabling the scattering properties of the protein and detergent to be investigated independently. The topological shape of LHC II, determined using ab initio shape restoration methods from the SANS data at the contrastmore » match point of BOG, was consistent with the X-ray crystallographic structure of LHC II (Liu et al. Nature 2004 428, 287-292). The interactions of the protein and detergent were investigated at the contrast match point for the protein and also in 100% D{sub 2}O. The data suggested that BOG micelle structure was altered by its interaction with LHC II, but large aggregate structures were not formed. Indirect Fourier transform analysis of the LHC II/BOG scattering curves showed that the increase in the maximum dimension of the protein-detergent complex was consistent with the presence of a monolayer of detergent surrounding the protein. A model of the LHC II/BOG complex was generated to interpret the measurements made in 100% D{sub 2}O. This model adequately reproduced the overall size of the LHC II/BOG complex, but demonstrated that the detergent does not have a highly regular shape that surrounds the hydrophobic periphery of LHC II. In addition to demonstrating that natively structured LHC II can be produced for functional characterization and for use in artificial solar energy applications, the analysis and modeling approaches described here can be used for characterizing detergent-associated {alpha}-helical transmembrane proteins.« less

  2. Development of an Efficient Protein Extraction Method Compatible with LC-MS/MS for Proteome Mapping in Two Australian Seagrasses Zostera muelleri and Posidonia australis.

    PubMed

    Jiang, Zhijian; Kumar, Manoj; Padula, Matthew P; Pernice, Mathieu; Kahlke, Tim; Kim, Mikael; Ralph, Peter J

    2017-01-01

    The availability of the first complete genome sequence of the marine flowering plant Zostera marina (commonly known as seagrass) in early 2016, is expected to significantly raise the impact of seagrass proteomics. Seagrasses are marine ecosystem engineers that are currently declining worldwide at an alarming rate due to both natural and anthropogenic disturbances. Seagrasses (especially species of the genus Zostera ) are compromised for proteomic studies primarily due to the lack of efficient protein extraction methods because of their recalcitrant cell wall which is rich in complex polysaccharides and a high abundance of secondary metabolites in their cells. In the present study, three protein extraction methods that are commonly used in plant proteomics i.e., phenol (P); trichloroacetic acid/acetone/SDS/phenol (TASP); and borax/polyvinyl-polypyrrolidone/phenol (BPP) extraction, were evaluated quantitatively and qualitatively based on two dimensional isoelectric focusing (2D-IEF) maps and LC-MS/MS analysis using the two most abundant Australian seagrass species, namely Zostera muelleri and Posidonia australis . All three tested methods produced high quality protein extracts with excellent 2D-IEF maps in P. australis . However, the BPP method produces better results in Z. muelleri compared to TASP and P. Therefore, we further modified the BPP method (M-BPP) by homogenizing the tissue in a modified protein extraction buffer containing both ionic and non-ionic detergents (0.5% SDS; 1.5% Triton X-100), 2% PVPP and protease inhibitors. Further, the extracted proteins were solubilized in 0.5% of zwitterionic detergent (C7BzO) instead of 4% CHAPS. This slight modification to the BPP method resulted in a higher protein yield, and good quality 2-DE maps with a higher number of protein spots in both the tested seagrasses. Further, the M-BPP method was successfully utilized in western-blot analysis of phosphoenolpyruvate carboxylase (PEPC-a key enzyme for carbon metabolism

  3. Isomeric Detergent Comparison for Membrane Protein Stability: Importance of Inter-Alkyl-Chain Distance and Alkyl Chain Length

    PubMed Central

    Cho, Kyung Ho; Hariharan, Parameswaran; Mortensen, Jonas S.; Du, Yang; Nielsen, Anne K.; Byrne, Bernadette; Kobilka, Brian K.; Loland, Claus J.; Guan, Lan

    2017-01-01

    Membrane proteins encapsulated by detergent micelles are widely used for structural study. Because of their amphipathic property, detergents have the ability to maintain protein solubility and stability in an aqueous medium. However, conventional detergents have serious limitations in their scope and utility, particularly for eukaryotic membrane proteins and membrane protein complexes. Thus, a number of new agents have been devised; some have made significant contributions to membrane protein structural studies. However, few detergent design principles are available. In this study, we prepared meta and ortho isomers of the previously reported para-substituted xylene-linked maltoside amphiphiles (XMAs), along with alkyl chain-length variation. The isomeric XMAs were assessed with three membrane proteins, and the meta isomer with a C12 alkyl chain was most effective at maintaining solubility/stability of the membrane proteins. We propose that interplay between the hydrophile–lipophile balance (HLB) and alkyl chain length is of central importance for high detergent efficacy. In addition, differences in inter-alkyl-chain distance between the isomers influence the ability of the detergents to stabilise membrane proteins. PMID:27981750

  4. Detection and quantification of anionic detergent (lissapol) in milk using attenuated total reflectance-Fourier Transform Infrared spectroscopy.

    PubMed

    Jaiswal, Pranita; Jha, Shyam Narayan; Kaur, Jaspreet; Borah, Anjan

    2017-04-15

    Adulteration of milk to gain economic benefit is rampant. Addition of detergent in milk can cause food poisoning and other complications. Fourier Transform Infrared spectroscopy was evaluated as rapid method for detection and quantification of anionic detergent (lissapol) in milk. Spectra of pure and artificially adulterated milk (0.2-2.0% detergent) samples revealed clear differences in wavenumber range of 4000-500cm -1 . The apparent variations observed in region of 1600-995 and 3040-2851cm -1 corresponds to absorption frequencies of common constituents of detergent (linear alkyl benzene sulphonate). Principal component analysis showed discrete clustering of samples based on level of detergent (p⩽0.05) in milk. The classification efficiency for test samples were recorded to be >93% using Soft Independent Modelling of Class Analogy approach. Maximum coefficient of determination for prediction of detergent was 0.94 for calibration and 0.93 for validation, using partial least square regression in wavenumber combination of 1086-1056, 1343-1333, 1507-1456, 3040-2851cm -1 . Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Chiral separation and quantitation of cetirizine and hydroxyzine by maltodextrin-mediated CE in human plasma: effect of zwitterionic property of cetirizine on enantioseparation.

    PubMed

    Nojavan, Saeed; Fakhari, Ali Reza

    2011-03-01

    In the present study, a very simple CE method for chiral separation and quantitation of zwitterionic cetirizine (CTZ), as the main metabolite of hydroxyzine (HZ), and HZ has been developed. In addition, the effect of zwitterionic property of CTZ on enantioseparation was investigated. Maltodextrin, a linear polysaccharide, as a chiral selector was used and several parameters affecting the separation such as pH of BGE, concentration of chiral selector and applied voltage were studied. The best BGE conditions for CTZ and HZ enantiomers were optimized as 75 mM sodium phosphate solution at pH of 2.0, containing 5% w/v maltodextrin. Results showed that, compared to HZ, pH of BGE was an effective parameter in enantioseparation of CTZ due to the zwitterionic property of CTZ. The linear range of the method was over 30-1200 ng/mL for all enantiomers of CTZ and HZ. The quantification and detection limits (S/N=3) of all enantiomers were 30 and 10 ng/mL, respectively. The method was used to quantitative enantioseparation of CTZ and HZ in spiked human plasma. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Synthetic detergents induced-biochemical and histological changes in skin of guinea pigs.

    PubMed

    Agarwal, C; Mathur, A K; Gupta, B N; Singh, A; Shanker, R

    1990-06-01

    The linear alkylbenzene sulphonate (LAS) based synthetic detergents-induced decrease in lipid peroxydation and increase in histamine content in exposed skin of guinea pigs in a dose-dependent manner. Histopathological alterations of exposed skin included moderate degree of hyperkeratinization at lower concentration but necrosis, scarring, sloughing as well as discontinuity of epidermis at higher concentrations. The results shows that the contact of skin with detergents causes dermal toxicity.

  7. Optimisation of saponin extraction conditions with Camellia sinensis var. assamica seed and its application for a natural detergent.

    PubMed

    Gong, Wanying; Huang, Yewei; Ji, Aibing; Peng, Wenshu; Liu, Cong; Zeng, Yin; Yang, Ruijuan; Yan, Liang; Wang, Xuanjun; Sheng, Jun

    2018-04-01

    Camellia sinensis var. assamica seed cake (a by-product of tea-seed oil) is an abundant resource with poor utilisation. C. sinensis var. assamica seed saponin (CSS) is one kind of non-ionic surfactant. In this study, the CSS extraction conditions were optimised by response surface methodology (RSM) and then the CSS detergent was developed. Additionally, the safety and decontamination ability of the developed detergent were evaluated. The optimised extraction conditions were including the extracting temperature of 40.04 °C, extraction time of 4.97 h, ethanol concentration of 64.11% and liquid-solid ratio of 14.57:1 mL g -1 . The formula of the CSS detergent was as follows: 20% crude CSS, 0.3% oxidised tea polyphenols (OTPs), 0.2% nisin, 0.3% sodium dehydroacetate, 0.7% sodium alginate and 0.5% sodium polyacrylate. The LD 50 of the CSS detergent exceeds 14 g kg -1 in mice, indicating the detergent was non-toxic. Both of the emulsifying and the pesticide residues removal abilities of the CSS detergent were significantly stronger than the commercial detergent. A natural tea seed saponin detergent with good safety and decontamination ability was successfully developed. This can make better use of the tea seed cake, thereby creating added value in the tea seed oil industry. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  8. Zwitterion mediator/quenchers. Coulombic minimization of the back-reaction in photocatalysis

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

    Brugger, P.A.; Graetzel, M.; Guarr, T.

    1982-03-18

    Zwitterionic analogues of methyl viologen, N,N'-bis(carboxyethyl)-4,4'-bipyridyl (CEB), and N,N'-bis(sulfonatopropyl)-4,4'-bipyridyl (SPB) have been studied as electron-transfer quenchers of excited-state photocatalysts. The molecules are formally neutral when oxidized but become negatively charged when reduced. This charge buildup minimizes undesirable back-recombination of the photogenerated redox products. This Coulombic effect is demonstrated both by direct flash photolysis measurements of back-recombination and by catalytic studies of water reduction. Results are compared for the zwitterion quenchers and methyl viologen, reacting with soluble anionic porphyrins and with micelle-associated reactants. For the anionic porphyrin system zinc tetrakis (sulfonatophenyl)porphyrin(4-) the rates of back-recombination are as follows: ZnTSPP/sup 3 -/more » + MV/sup +/...-->.. ZnTSPP/sup 4 -/ + MV/sup 2 +/ (k), k = 2 x 10/sup 9/M/sup -1/s/sup -1/; ZnTSPP/sup 3 -/ + CEB/sup -/. ..-->.. ZnTSPP/sup 4 -/ + CEB +- (k), k = 6 x 10/sup 8/ M/sup -1/ s/sup -1/. In a corresponding catalytic system incorporating ZnTPPS as the photoacceptor, CEB or methyl viologen as the quencher, and N-phenylglycine as the electron donor, a >2-fold enhancement of catalysis is observed by replacing MV/sup 2 +/ with CEB. These events can be greatly accentuated by increasing the charge on the components, via micellar localization. Thus, by using an amphiphilic derivative of Ru(bpy)/sub 3//sup 2 +/ as photoactive donor solubilized in anionic micelles and SPB as electron acceptor, one can achieve 200-fold reduction in the rate of the back-reaction.« less

  9. 40 CFR 180.1022 - Iodine-detergent complex; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Iodine-detergent complex; exemption... FOOD Exemptions From Tolerances § 180.1022 Iodine-detergent complex; exemption from the requirement of a tolerance. The aqueous solution of hydriodic acid and elemental iodine, including one or both of...

  10. Detergent protease exposure and respiratory disease: case-referent analysis of a retrospective cohort.

    PubMed

    Brant, A; Upchurch, S; van Tongeren, M; Zekveld, C; Helm, J; Barnes, F; Newman Taylor, A J; Cullinan, P

    2009-11-01

    To examine the relationship between protease exposure and respiratory disease in a cohort of detergent enzyme manufacturers. Case-referent analysis of a cohort of employees working in a European detergent factory between 1989 and 2002. Cases with new lower or upper respiratory disease were ascertained by examination of occupational health records and matched to referents on date of first employment. Personal exposures to airborne detergent protease were estimated, using a job exposure matrix, from >12,000 measurements taken in the factory during the period of study. We found clear, monotonic relationships between estimated protease exposure and both lower and upper respiratory disease. After control for age, sex and smoking, the odds ratio of lower respiratory disease was significantly elevated (1.98, 95% CI 1.04 to 3.79) in those employees working in jobs in the highest quartile of protease exposure (geometric mean 7.9 ng x m(-3)). For employees with upper respiratory disease, the risk was significantly elevated at a lower level of estimated protease exposure (geometric mean 2.3 ng x m(-3)). These findings provide strong evidence of an association between detergent enzyme exposure and the development of respiratory disease in an occupational setting. Using the routinely collected information on specific sensitisation and the close attention to workplace exposures that are characteristic of this industry, it should be possible to derive meaningful occupational exposure standards for most detergent enzymes.

  11. Role of Detergents in Conformational Exchange of a G Protein-coupled Receptor*

    PubMed Central

    Chung, Ka Young; Kim, Tae Hun; Manglik, Aashish; Alvares, Rohan; Kobilka, Brian K.; Prosser, R. Scott

    2012-01-01

    The G protein-coupled β2-adrenoreceptor (β2AR) signals through the heterotrimeric G proteins Gs and Gi and β-arrestin. As such, the energy landscape of β2AR-excited state conformers is expected to be complex. Upon tagging Cys-265 of β2AR with a trifluoromethyl probe, 19F NMR was used to assess conformations and possible equilibria between states. Here, we report key differences in β2AR conformational dynamics associated with the detergents used to stabilize the receptor. In dodecyl maltoside (DDM) micelles, the spectra are well represented by a single Lorentzian line that shifts progressively downfield with activation by appropriate ligand. The results are consistent with interconversion between two or more states on a time scale faster than the greatest difference in ligand-dependent chemical shift (i.e. >100 Hz). Given that high detergent off-rates of DDM monomers may facilitate conformational exchange between functional states of β2AR, we utilized the recently developed maltose-neopentyl glycol (MNG-3) diacyl detergent. In MNG-3 micelles, spectra indicated at least three distinct states, the relative populations of which depended on ligand, whereas no ligand-dependent shifts were observed, consistent with the slow exchange limit. Thus, detergent has a profound effect on the equilibrium kinetics between functional states. MNG-3, which has a critical micelle concentration in the nanomolar regime, exhibits an off-rate that is 4 orders of magnitude lower than that of DDM. High detergent off-rates are more likely to facilitate conformational exchange between distinct functional states associated with the G protein-coupled receptor. PMID:22893704

  12. Role of detergents in conformational exchange of a G protein-coupled receptor.

    PubMed

    Chung, Ka Young; Kim, Tae Hun; Manglik, Aashish; Alvares, Rohan; Kobilka, Brian K; Prosser, R Scott

    2012-10-19

    The G protein-coupled β(2)-adrenoreceptor (β(2)AR) signals through the heterotrimeric G proteins G(s) and G(i) and β-arrestin. As such, the energy landscape of β(2)AR-excited state conformers is expected to be complex. Upon tagging Cys-265 of β(2)AR with a trifluoromethyl probe, (19)F NMR was used to assess conformations and possible equilibria between states. Here, we report key differences in β(2)AR conformational dynamics associated with the detergents used to stabilize the receptor. In dodecyl maltoside (DDM) micelles, the spectra are well represented by a single Lorentzian line that shifts progressively downfield with activation by appropriate ligand. The results are consistent with interconversion between two or more states on a time scale faster than the greatest difference in ligand-dependent chemical shift (i.e. >100 Hz). Given that high detergent off-rates of DDM monomers may facilitate conformational exchange between functional states of β(2)AR, we utilized the recently developed maltose-neopentyl glycol (MNG-3) diacyl detergent. In MNG-3 micelles, spectra indicated at least three distinct states, the relative populations of which depended on ligand, whereas no ligand-dependent shifts were observed, consistent with the slow exchange limit. Thus, detergent has a profound effect on the equilibrium kinetics between functional states. MNG-3, which has a critical micelle concentration in the nanomolar regime, exhibits an off-rate that is 4 orders of magnitude lower than that of DDM. High detergent off-rates are more likely to facilitate conformational exchange between distinct functional states associated with the G protein-coupled receptor.

  13. New Vary-Chap Profile of the Topside Ionosphere Electron Density Distribution for use with the IRI Model and the GIRO Real-Time Data

    NASA Technical Reports Server (NTRS)

    Nsumei, Patrick; Reinisch, Bodo W.; Huang, Xueqin; Bilitza, Dieter

    2012-01-01

    A new Vary-Chap function is introduced for the empirical modeling of the electron density N(h) profile in the topside ionosphere that uses a shape function S(h) in the generalized Chapman function. The Vary-Chap profile extends the bottomside profile that is specified by the IRI model or measured by the Global Ionospheric Radio Observatory (GIRO) to the altitude of the ISIS-2 satellite. Some 80,000 topside profiles, measured by the topside sounder on the ISIS-2 satellite were analyzed, and the shape function S(h) was calculated for each profile. A parameterized function S*(h), composed of two sub-functions S1(h) and S2(h), is fitted to the measured S(h) profile using three free parameters. At altitudes just above the F2 layer peak height hmF2, the shape function S1 controls S(h), and at greater altitudes S2 controls S(h). The height of the intersection of S1 and S2 is defined as the transition height h(sub T) indicating the transition from an O(+) to an H(+)-dominated profile shape. The observed transition heights range from approx.500 km to 800 km.

  14. Ion-Specific Modulation of Interfacial Interaction Potentials between Solid Substrates and Cell-Sized Particles Mediated via Zwitterionic, Super-Hydrophilic Poly(sulfobetaine) Brushes.

    PubMed

    Higaki, Yuji; Fröhlich, Benjamin; Yamamoto, Akihisa; Murakami, Ryo; Kaneko, Makoto; Takahara, Atsushi; Tanaka, Motomu

    2017-02-16

    Zwitterionic polymer brushes draw increasing attention not only because of their superhydrophilic, self-cleaning capability but also due to their excellent antifouling capacity. We investigated the ion-specific modulation of the interfacial interaction potential via densely packed, uniform poly(sulfobetaine) brushes. The vertical Brownian motion of a cell-sized latex particle was monitored by microinterferometry, yielding the effective interfacial interaction potentials V(Δh) and the autocorrelation function of height fluctuation. The potential curvature V″(Δh) exhibited a monotonic increase according to the increase in monovalent salt concentrations, implying the sharpening of the potential confinement. An opposite tendency was observed in CaCl 2 solutions, suggesting that the ion specific modulation cannot be explained by the classical Hofmeister series. When the particle fluctuation was monitored in the presence of free sulfobetaine molecules, the increase in [sulfobetaine] resulted in a distinct increase in hydrodynamic friction. This was never observed in all the other salt solutions, suggesting the interference of zwitterionic pairing of sulfobetaine side chains by the intercalation of sulfobetaine molecules into the brush layer. Furthermore, poly(sulfobetaine) brushes exhibited a very low V″(Δh) and hydrodynamic friction to human erythrocytes, which seems to explain the excellent blood repellency of zwitterionic polymer materials.

  15. 40 CFR 80.141 - Interim detergent gasoline program.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... considered acceptable for demonstration of IVD control performance. Examples of acceptable test procedures... carburetor deposits. Examples of acceptable test procedures for demonstration of carburetor deposit control... ultimate consumer; (ii) All additized post-refinery component (PRC); and (iii) All detergent additives sold...

  16. 40 CFR 80.141 - Interim detergent gasoline program.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... considered acceptable for demonstration of IVD control performance. Examples of acceptable test procedures... carburetor deposits. Examples of acceptable test procedures for demonstration of carburetor deposit control... ultimate consumer; (ii) All additized post-refinery component (PRC); and (iii) All detergent additives sold...

  17. 40 CFR 80.141 - Interim detergent gasoline program.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... considered acceptable for demonstration of IVD control performance. Examples of acceptable test procedures... carburetor deposits. Examples of acceptable test procedures for demonstration of carburetor deposit control... ultimate consumer; (ii) All additized post-refinery component (PRC); and (iii) All detergent additives sold...

  18. 40 CFR 80.141 - Interim detergent gasoline program.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... considered acceptable for demonstration of IVD control performance. Examples of acceptable test procedures... carburetor deposits. Examples of acceptable test procedures for demonstration of carburetor deposit control... ultimate consumer; (ii) All additized post-refinery component (PRC); and (iii) All detergent additives sold...

  19. Detergent-dispersant additives based on high-molecular-weight alkylphenols

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

    Kulieva, K.N.; Namazova, I.I.; Ismailova, N.D.

    1988-09-01

    This article describes the synthesis and investigation of Mannich bases produced for alkylphenols, obtained in turn from ethylene oligomers. These oligomers are the still bottoms from distillation products of high-temperature oligomerization of ethylene in the presence of triethylaluminum. Two narrow cuts obtained from the distillation of oligomer fraction were used to study the influence of ethylene oligomer molecular weight on the properties of the additives. The additives were blended in DS-11 oil to evaluate their detergency-dispersancy and other properties. Comparison blends were made with succinimide additives based on the same ethylene oligomers. The Mannich bases give improvements in the oxidationmore » resistance, anticorrosion properties, and detergency-dispersancy of the DS-11 diesel oil.« less

  20. Occupational asthma and allergy in the detergent industry: new developments.

    PubMed

    Sarlo, Katherine; Kirchner, Donald B

    2002-04-01

    This review highlights the latest developments in the control of enzyme-induced occupational asthma and allergy (rhinitis and conjunctivitis) in the detergent industry. The industry has developed guidelines for the safe handling of enzymes in order to reduce the risk of occupational allergy and asthma. Those manufacturing facilities that follow all of the guidelines enjoy very low or no cases of asthma and allergy among workers exposed to enzymes. The key to the success of the management of enzyme-induced allergy and asthma is prospective surveillance for the development of enzyme-specific IgE antibody before the onset of allergic symptoms. This allows for continuing interventions to reduce exposures, so as to minimize or eliminate those associated with symptoms. Workers with IgE to enzymes can still continue to work in the industry symptom-free for their entire career. This indicates that exposures needed to induce sensitization are different and probably lower than exposures needed to elicit enzyme allergic symptoms. The experience of the detergent enzyme industry in controlling occupational allergens can be applied to other industries. The detergent enzyme story can be viewed as a model for the control of type 1 protein allergens in the workplace.

  1. Investigation of the photophysical and photochemical properties of peripherally tetra-substituted water-soluble zwitterionic and cationic zinc(ii) phthalocyanines.

    PubMed

    Çolak, Senem; Durmuş, Mahmut; Yıldız, Salih Zeki

    2016-06-21

    In this study, 4-{4-[N-((3-dimethylamino)propyl)amide]phenoxy}phthalonitrile () and its zinc(ii) phthalocyanine derivative () were synthesized for the first time. 4-(N-((3-Dimethylamino)propyl)amide)phenoxy substituted zinc(ii) phthalocyanine () was converted to its water-soluble sulfobetaine (), betaine () and N-oxide () containing zwitterionic and quaternized cationic () derivatives. All newly synthesized compounds () were characterized by the combination of UV-vis, FT-IR, (1)H NMR, mass spectroscopy techniques and elemental analysis. The photophysical (fluorescence quantum yields and lifetimes) and photochemical (singlet oxygen quantum yields) properties were investigated in DMSO for all the synthesized zinc(ii) phthalocyanines () and in both DMSO and aqueous solutions for zwitterionic and cationic phthalocyanines () for the specification of their capability as photosensitizers in photodynamic therapy (PDT). The binding behavior of water soluble phthalocyanines () to the bovine serum albumin protein was also examined for the determination of their transportation ability in the blood stream.

  2. Isomeric Detergent Comparison for Membrane Protein Stability: Importance of Inter-Alkyl-Chain Distance and Alkyl Chain Length.

    PubMed

    Cho, Kyung Ho; Hariharan, Parameswaran; Mortensen, Jonas S; Du, Yang; Nielsen, Anne K; Byrne, Bernadette; Kobilka, Brian K; Loland, Claus J; Guan, Lan; Chae, Pil Seok

    2016-12-14

    Membrane proteins encapsulated by detergent micelles are widely used for structural study. Because of their amphipathic property, detergents have the ability to maintain protein solubility and stability in an aqueous medium. However, conventional detergents have serious limitations in their scope and utility, particularly for eukaryotic membrane proteins and membrane protein complexes. Thus, a number of new agents have been devised; some have made significant contributions to membrane protein structural studies. However, few detergent design principles are available. In this study, we prepared meta and ortho isomers of the previously reported para-substituted xylene-linked maltoside amphiphiles (XMAs), along with alkyl chain-length variation. The isomeric XMAs were assessed with three membrane proteins, and the meta isomer with a C 12 alkyl chain was most effective at maintaining solubility/stability of the membrane proteins. We propose that interplay between the hydrophile-lipophile balance (HLB) and alkyl chain length is of central importance for high detergent efficacy. In addition, differences in inter-alkyl-chain distance between the isomers influence the ability of the detergents to stabilise membrane proteins. © 2016 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Recovery and redispersion of gold nanoparticles using the self-assembly of a pH sensitive zwitterionic amphiphile.

    PubMed

    Morita-Imura, Clara; Imura, Yoshiro; Kawai, Takeshi; Shindo, Hitoshi

    2014-11-04

    The pH-responsive self-assembly of zwitterionic amphiphile C16CA was expanded to the recovery of gold (Au) nanoparticles for environmentally friendly chemistry applications. Multilayered lamellae at pH ∼ 4 were successfully incorporated into nanoparticles by dispersion. Redispersion of nanoparticles was achieved under basic conditions by the transition of self-assembly.

  4. Effect of Detergents on Galactoside Binding by Melibiose Permeases.

    PubMed

    Amin, Anowarul; Hariharan, Parameswaran; Chae, Pil Seok; Guan, Lan

    2015-09-29

    The effect of various detergents on the stability and function of the melibiose permeases of Escherichia coli (MelBEc) and Salmonella typhimurium (MelBSt) was studied. In n-dodecyl-β-d-maltoside (DDM) or n-undecyl-β-d-maltoside (UDM), WT MelBSt binds melibiose with an affinity similar to that in the membrane. However, with WT MelBEc or MelBSt mutants (Arg141 → Cys, Arg295 → Cys, or Arg363 → Cys), galactoside binding is not detected in these detergents, but binding to the phosphotransferase protein IIA(Glc) is maintained. In the amphiphiles lauryl maltose neopentyl glycol (MNG-3) or glyco-diosgenin (GDN), galactoside binding with all of the MelB proteins is observed, with slightly reduced affinities. MelBSt is more thermostable than MelBEc, and the thermostability of either MelB is largely increased in MNG-3 or GDN. Therefore, the functional defect with DDM or UDM likely results from the relative instability of the sensitive MelB proteins, and stability, as well as galactoside binding, is retained in MNG-3 or GDN. Furthermore, isothermal titration calorimetry of melibiose binding with MelBSt shows that the favorable entropic contribution to the binding free energy is decreased in MNG-3, indicating that the conformational dynamics of MelB is restricted in this detergent.

  5. Effect of detergents on galactoside binding by melibiose permeases

    PubMed Central

    Amin, Anowarul; Hariharan, Parameswaran; Chae, Pil Seok; Guan, Lan

    2015-01-01

    The effect of various detergents on the stability and function of melibiose permeases of Escherichia coli (MelBEc) or Salmonella typhimurium (MelBSt) were studied. In n-dodecyl-β-d-maltoside (DDM) or n-undecyl-β-d-maltoside (UDM), WT MelBSt binds melibiose with an affinity similar to that in the membrane. However, with WT MelBEc or MelBSt mutants (Arg141→Cys, Arg295→Cys or Arg363→Cys), galactoside binding is not detected in these detergents, but binding to the phosphotransferase protein IIAGlc is maintained. In the amphiphiles lauryl maltose neopentyl glycol (MNG-3) or glyco-diosgenin (GDN), galactoside binding with all the MelB proteins is observed, with slightly reduced affinities. MelBSt is more thermostable than MelBEc, and the thermostability of either MelB is largely increased in MNG-3 or GDN. Therefore, the functional defect with DDM or UDM likely results from relative instability of the sensitive MelB proteins, and stability, as well as galactoside binding, is retained in MNG-3 or GDN. Furthermore, isothermal titration calorimetry of melibiose binding with MelBSt shows that the favorable entropic contribution to the binding free energy is decreased in MNG-3, indicating that the conformational dynamics of MelB is restricted in this detergent. PMID:26352464

  6. Differential effects of non-ionic detergents on microsomal and sarcolemmal adenylate cyclase in cardiac muscle

    PubMed Central

    Sulakhe, Prakash V.; Narayanan, Njanoor

    1978-01-01

    1. About 4 and 23% of the homogenate adenylate cyclase activity was recovered in the microsomal and sarcolemmal fractions isolated from guinea-pig heart ventricles. 2. Cardiac microsomal adenylate cyclase activity [basal as well as p[NH]ppG (guanyl-5′-yl imidodiphosphate)- and NaF-stimulated] was increased over 2-fold in the presence of Lubrol-PX (0.01–0.1%). 3. The sarcolemmal enzyme, however, showed concentration-dependent inhibition caused by the detergent under all assay conditions, except when p[NH]ppG was included in the assay. In the latter case, the detergent (0.01–0.02%) caused a modest increase (30–45%) in enzyme activity. 4. Another non-ionic detergent, Triton X-100, also stimulated the microsomal cyclase and inhibited the sarcolemmal enzyme. 5. With either membrane fraction, Lubrol-PX solubilized the enzyme when the detergent/membrane protein ratio was 2.5 (μmol of detergent/mg of protein). 6. The findings with homogenate and a washed particulate fraction resembled those obtained with sarcolemma, and those with isolated sarcoplasmic reticulum resembled those with microsomal preparations. 7. p[NH]ppG, and to some extent NaF, protected the detergent-induced inactivation of the enzyme observed at higher detergent concentrations (0.5% Lubrol-PX and 0.05–0.5% Triton X-100). 8. In the absence of detergents, p[NH]ppG increased the basal enzyme activity about 2-fold in microsomal fractions, but did not appreciably stimulate the sarcolemmal enzyme. Isoproterenol, on the other hand, increased the sarcolemmal enzyme activity (>2-fold) in the presence of p[NH]ppG and caused only moderate stimulation (31%) of the microsomal enzyme under these conditions. 9. These findings support the view that, although the bulk of adenylate cyclase resides in heart sarcolemma (plasma membrane), the microsomal activity cannot be accounted for solely by contamination of the microsomal fraction with sarcolemma, as has been suggested by others [Besch, Jones & Watanabe (1976

  7. Toxicity assessment of a common laundry detergent using the freshwater flagellate Euglena gracilis.

    PubMed

    Azizullah, Azizullah; Richter, Peter; Häder, Donat-Peter

    2011-09-01

    Synthetic detergents are among the commonly used chemicals in everyday life. Detergents, reaching aquatic environments through domestic and municipal wastewater, can cause many different effects in aquatic organisms. The present study was aimed at the toxicity evaluation of a commonly used laundry detergent, Ariel, using the freshwater flagellate Euglena gracilis as a biotest organism. Different parameters of the flagellate like motility, swimming velocity, cell shape, gravitactic orientation, photosynthesis and concentration of light harvesting pigments were used as end points for the toxicity assessment. No Observed Effect Concentration (NOEC) and EC(50) values were calculated for the end point parameters at four different incubation times, i.e. 0, 6, 24 and 72 h. After 72 h incubation, swimming velocity of the cells was found to be the most sensitive parameter giving NOEC and EC(50) values of 10.8 and 34 mg L(-1), respectively. After 72 h exposure to the detergent, chlorophyll a and total carotenoids were significantly decreased in cultures treated with Ariel at concentrations of 50 mg L(-1) and above while chlorophyll b significantly decreased at concentrations above 750 mg L(-1). The maximum inhibitory effect on the quantum yield of photosystem II was observed after 24 h exposure and thereafter a recovery trend was observed. Motility, gravitaxis and cell shape were strongly impaired immediately upon exposure to the detergent, but with increasing exposure time these parameters showed acclimatization to the stress and thus the NOEC values obtained after 72 h were higher than those immediately after exposure. Copyright © 2011 Elsevier Ltd. All rights reserved.

  8. Lipid domains in zwitterionic-anionic lipid mixtures induced by combined effect of monovalent and divalent ions

    NASA Astrophysics Data System (ADS)

    Xu, Hongcheng; Ganesan, Sai; Matysiak, Silvina

    Lipid domain formation is an important process for many cellular processes. In experiment, the effects of Ba2+, Sr2+, Ca2+ and Mg2+ in inducing lateral phase separation in the binary phosphatidylcholine-phosphatidylserine (PC-PS) bilayer are quite different, of which the molecular mechanism remains to be understood. We have explored the effect of monovalent (MI) and divalent (MII) cationic radii on lipid domain formation in mixed zwitterionic-anionic lipid bilayers. We propose a mechanism for the formation of divalent-cation-induced lipid domains based on MD simulations with our Water-Explicit Polarizable MEMbrane (WEPMEM) coarse-grained model, which uses PC as the model for zwitterionic and PS for anionic lipids. Lipid aggregation only occurs with limited range of monovalent and divalent ion sizes in agreement with experimental observations. More ordering and closer packing of the lipids are noted within the domains, which correlate with bilayer thickness, curvature and lipid asymmetry. The results of the simulations reveal that the lipid domain consists of MII-mediated anionic lipid dimer/trimer complexes bridged by monovalent ions MI and provide a stereochemical insight in understanding the experimentally observed calcium-induced phase separation.

  9. Highly Specific Binding on Antifouling Zwitterionic Polymer-Coated Microbeads as Measured by Flow Cytometry.

    PubMed

    van Andel, Esther; de Bus, Ian; Tijhaar, Edwin J; Smulders, Maarten M J; Savelkoul, Huub F J; Zuilhof, Han

    2017-11-08

    Micron- and nano-sized particles are extensively used in various biomedical applications. However, their performance is often drastically hampered by the nonspecific adsorption of biomolecules, a process called biofouling, which can cause false-positive and false-negative outcomes in diagnostic tests. Although antifouling coatings have been extensively studied on flat surfaces, their use on micro- and nanoparticles remains largely unexplored, despite the widespread experimental (specifically, clinical) uncertainties that arise because of biofouling. Here, we describe the preparation of magnetic micron-sized beads coated with zwitterionic sulfobetaine polymer brushes that display strong antifouling characteristics. These coated beads can then be equipped with recognition elements of choice, to enable the specific binding of target molecules. First, we present a proof of principle with biotin-functionalized beads that are able to specifically bind fluorescently labeled streptavidin from a complex mixture of serum proteins. Moreover, we show the versatility of the method by demonstrating that it is also possible to functionalize the beads with mannose moieties to specifically bind the carbohydrate-binding protein concanavalin A. Flow cytometry was used to show that thus-modified beads only bind specifically targeted proteins, with minimal/near-zero nonspecific protein adsorption from other proteins that are present. These antifouling zwitterionic polymer-coated beads, therefore, provide a significant advancement for the many bead-based diagnostic and other biosensing applications that require stringent antifouling conditions.

  10. Highly Specific Binding on Antifouling Zwitterionic Polymer-Coated Microbeads as Measured by Flow Cytometry

    PubMed Central

    2017-01-01

    Micron- and nano-sized particles are extensively used in various biomedical applications. However, their performance is often drastically hampered by the nonspecific adsorption of biomolecules, a process called biofouling, which can cause false-positive and false-negative outcomes in diagnostic tests. Although antifouling coatings have been extensively studied on flat surfaces, their use on micro- and nanoparticles remains largely unexplored, despite the widespread experimental (specifically, clinical) uncertainties that arise because of biofouling. Here, we describe the preparation of magnetic micron-sized beads coated with zwitterionic sulfobetaine polymer brushes that display strong antifouling characteristics. These coated beads can then be equipped with recognition elements of choice, to enable the specific binding of target molecules. First, we present a proof of principle with biotin-functionalized beads that are able to specifically bind fluorescently labeled streptavidin from a complex mixture of serum proteins. Moreover, we show the versatility of the method by demonstrating that it is also possible to functionalize the beads with mannose moieties to specifically bind the carbohydrate-binding protein concanavalin A. Flow cytometry was used to show that thus-modified beads only bind specifically targeted proteins, with minimal/near-zero nonspecific protein adsorption from other proteins that are present. These antifouling zwitterionic polymer-coated beads, therefore, provide a significant advancement for the many bead-based diagnostic and other biosensing applications that require stringent antifouling conditions. PMID:29064669

  11. Detergent Optimized Membrane Protein Reconstitution in Liposomes for Solid State NMR

    PubMed Central

    2015-01-01

    For small helical membrane proteins, their structures are highly sensitive to their environment, and solid state NMR is a structural technique that can characterize these membrane proteins in native-like lipid bilayers and proteoliposomes. To date, a systematic method by which to evaluate the effect of the solubilizing detergent on proteoliposome preparations for solid state NMR of membrane proteins has not been presented in the literature. A set of experiments are presented aimed at determining the conditions most amenable to dialysis mediated reconstitution sample preparation. A membrane protein from M. tuberculosis is used to illustrate the method. The results show that a detergent that stabilizes the most protein is not always ideal and sometimes cannot be removed by dialysis. By focusing on the lipid and protein binding properties of the detergent, proteoliposome preparations can be readily produced, which provide double the signal-to-noise ratios for both the oriented sample and magic angle spinning solid state NMR. The method will allow more membrane protein drug targets to be structurally characterized in lipid bilayer environments. PMID:24665863

  12. Effects of age, season, gender and urban-rural status on time-activity: CanadianHuman Activity Pattern Survey 2 (CHAPS 2).

    PubMed

    Matz, Carlyn J; Stieb, David M; Davis, Karelyn; Egyed, Marika; Rose, Andreas; Chou, Benedito; Brion, Orly

    2014-02-19

    Estimation of population exposure is a main component of human health risk assessment for environmental contaminants. Population-level exposure assessments require time-activity pattern distributions in relation to microenvironments where people spend their time. Societal trends may have influenced time-activity patterns since previous Canadian data were collected 15 years ago. The Canadian Human Activity Pattern Survey 2 (CHAPS 2) was a national survey conducted in 2010-2011 to collect time-activity information from Canadians of all ages. Five urban and two rural locations were sampled using telephone surveys. Infants and children, key groups in risk assessment activities, were over-sampled. Survey participants (n = 5,011) provided time-activity information in 24-hour recall diaries and responded to supplemental questionnaires concerning potential exposures to specific pollutants, dwelling characteristics, and socio-economic factors. Results indicated that a majority of the time was spent indoors (88.9%), most of which was indoors at home, with limited time spent outdoors (5.8%) or in a vehicle (5.3%). Season, age, gender and rurality were significant predictors of time activity patterns. Compared to earlier data, adults reported spending more time indoors at home and adolescents reported spending less time outdoors, which could be indicative of broader societal trends. These findings have potentially important implications for assessment of exposure and risk. The CHAPS 2 data also provide much larger sample sizes to allow for improved precision and are more representative of infants, children and rural residents.

  13. Effects of Age, Season, Gender and Urban-Rural Status on Time-Activity: Canadian Human Activity Pattern Survey 2 (CHAPS 2)

    PubMed Central

    Matz, Carlyn J.; Stieb, David M.; Davis, Karelyn; Egyed, Marika; Rose, Andreas; Chou, Benedito; Brion, Orly

    2014-01-01

    Estimation of population exposure is a main component of human health risk assessment for environmental contaminants. Population-level exposure assessments require time-activity pattern distributions in relation to microenvironments where people spend their time. Societal trends may have influenced time-activity patterns since previous Canadian data were collected 15 years ago. The Canadian Human Activity Pattern Survey 2 (CHAPS 2) was a national survey conducted in 2010–2011 to collect time-activity information from Canadians of all ages. Five urban and two rural locations were sampled using telephone surveys. Infants and children, key groups in risk assessment activities, were over-sampled. Survey participants (n = 5,011) provided time-activity information in 24-hour recall diaries and responded to supplemental questionnaires concerning potential exposures to specific pollutants, dwelling characteristics, and socio-economic factors. Results indicated that a majority of the time was spent indoors (88.9%), most of which was indoors at home, with limited time spent outdoors (5.8%) or in a vehicle (5.3%). Season, age, gender and rurality were significant predictors of time activity patterns. Compared to earlier data, adults reported spending more time indoors at home and adolescents reported spending less time outdoors, which could be indicative of broader societal trends. These findings have potentially important implications for assessment of exposure and risk. The CHAPS 2 data also provide much larger sample sizes to allow for improved precision and are more representative of infants, children and rural residents. PMID:24557523

  14. Poly(zwitterionic liquids) functionalized polypyrrole/graphene oxide nanosheets for electrochemically detecting dopamine at low concentration.

    PubMed

    Mao, Hui; Liang, Jiachen; Ji, Chunguang; Zhang, Haifeng; Pei, Qi; Zhang, Yuyang; Zhang, Yu; Hisaeda, Yoshio; Song, Xi-Ming

    2016-08-01

    Poly(3-(1-vinylimidazolium-3-yl)propane-1-sulfonate) (PVIPS), a novel kind of poly(zwitterionic liquids) (PZILs) containing both imidazolium cation and sulfonate anion, was successfully modified on the surface of polypyrrole/graphene oxide nanosheets (PPy/GO) by covalent bonding. The obtained novel PZILs functionalized PPy/GO nanosheets (PVIPS/PPy/GO) modified glassy carbon electrode (GCE) presented the excellent electrochemical catalytic activity towards dopamine (DA) with high stability, sensitivity, selectivity and wide linear range (40-1220nM), especially having a lower detection limit (17.3nM). The excellent analytical performance is attributed to the strongly negative charges on the surface of modified GCE in aqueous solution, which is different from conventional poly(ionic liquids) modified GCE. DA cations could be quickly enriched on the electrode surface by electrostatic interaction in solution due to the existence of SO3(-) groups with negative charge at the end of pendant groups in zwitterionic PVIPS, resulting in a change of the electrons transmission mode in the oxidation of DA, that is, from a typical diffusion-controlled process at conventional poly(1-vinyl-3-ethylimidazole bromide) (PVEIB)/PPy/GO modified GCE to a typical surface-controlled process. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. 40 CFR 80.169 - Liability for violations of the detergent certification program controls and prohibitions.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...-additized PRC satisfied relevant requirements when it left the party's control; and (iii) That the party has... satisfied all relevant requirements when it left the detergent manufacturer's control. (B) Written blending... was manufactured. (2) To establish that, when it left the manufacturer's control, the detergent...

  16. Enhancing the specificity of polymerase chain reaction by graphene oxide through surface modification: zwitterionic polymer is superior to other polymers with different charges.

    PubMed

    Zhong, Yong; Huang, Lihong; Zhang, Zhisen; Xiong, Yunjing; Sun, Liping; Weng, Jian

    Graphene oxides (GOs) with different surface characteristics, such as size, reduction degree and charge, are prepared, and their effects on the specificity of polymerase chain reaction (PCR) are investigated. In this study, we demonstrate that GO with a large size and high reduction degree is superior to small and nonreduced GO in enhancing the specificity of PCR. Negatively charged polyacrylic acid (PAA), positively charged polyacrylamide (PAM), neutral polyethylene glycol (PEG) and zwitterionic polymer poly(sulfobetaine) (pSB) are used to modify GO. The PCR specificity-enhancing ability increases in the following order: GO-PAA < GO-PAM < GO-PEG < GO-pSB. Thus, zwitterionic polymer-modified GO is superior to other GO derivatives with different charges in enhancing the specificity of PCR. GO derivatives are also successfully used to enhance the specificity of PCR for the amplification of human mitochondrial DNA using blood genomic DNA as template. Molecular dynamics simulations and molecular docking are performed to elucidate the interaction between the polymers and Pfu DNA polymerase. Our data demonstrate that the size, reduction degree and surface charge of GO affect the specificity of PCR. Based on our results, zwitterionic polymer-modified GO may be used as an efficient additive for enhancing the specificity of PCR.

  17. Memprot: a program to model the detergent corona around a membrane protein based on SEC–SAXS data

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

    Pérez, Javier, E-mail: javier.perez@synchrotron-soleil.fr; Koutsioubas, Alexandros; Synchrotron SOLEIL, L’Orme des Merisiers, BP 48, Saint-Aubin, 91192 Gif-sur-Yvette

    Systematic SAXS simulations have been analysed over a wide range of parameters in order to better understand the detergent corona around a membrane protein. The application of small-angle X-ray scattering (SAXS) to structural investigations of transmembrane proteins in detergent solution has been hampered by two main inherent hurdles. On the one hand, the formation of a detergent corona around the hydrophobic region of the protein strongly modifies the scattering curve of the protein. On the other hand, free micelles of detergent without a precisely known concentration coexist with the protein–detergent complex in solution, therefore adding an uncontrolled signal. To gainmore » robust structural information on such systems from SAXS data, in previous work, advantage was taken of the online combination of size-exclusion chromatography (SEC) and SAXS, and the detergent corona around aquaporin-0, a membrane protein of known structure, could be modelled. A precise geometrical model of the corona, shaped as an elliptical torus, was determined. Here, in order to better understand the correlations between the corona model parameters and to discuss the uniqueness of the model, this work was revisited by analyzing systematic SAXS simulations over a wide range of parameters of the torus.« less

  18. Proteolytic Enzymes in Detergents: Evidence of Their Presence through Activity Measurements Based on Electrophoresis

    ERIC Educational Resources Information Center

    Saperas, Nuria; Fonfria-Subiros, Elsa

    2011-01-01

    This laboratory exercise uses a problem-based approach to expose students to some basic concepts relating to proteins and enzymes. One of the main applications of enzymes at the industrial level is their use in the detergent market. The students examine a detergent sample to ascertain whether proteolytic enzymes are a component and, if so, which…

  19. Technical note: improved methodology for analyses of acid detergent fiber and acid detergent lignin.

    PubMed

    Raffrenato, E; Van Amburgh, M E

    2011-07-01

    The objective of this study was to evaluate the methodology of the acid detergent lignin (ADL) assay in an effort to evaluate particle loss, improve repeatability, and decrease variation within and among samples. The original ADL method relied on asbestos as a filtering aid, but that was removed in 1989 with the mandate from the Environmental Protection Agency to eliminate asbestos in the environment. Furthermore, recent work on fiber methodology indicated that pore size in the Gooch sintered glass crucible (40-60 μm) was too large to trap all of the small particles associated with neutral detergent fiber (NDF) and acid detergent fiber (ADF). Thus, any loss of ADF could potentially result in a loss of ADL. Sixty forages including conventional and brown midrib corn silages, alfalfa silages and hays, mature grasses, early vegetative grasses, and 9 feces samples, were analyzed sequentially for ADF and ADL as outlined in the 1973 procedure of Van Soest except for the use of the asbestos fiber. A glass microfiber filter with a 1.5-μm pore size was chosen as a filtering aid because it met the criteria required by the assay: glass, heat resistant, acid resistant, chemically inert, and hydrophobic. To compare with the current ADF and ADL assays, the assays were conducted with either no filter or the glass filter inserted into crucibles, rinsed with acetone, and then according to the 1973 procedure of Van Soest. The samples analyzed covered a range from 18.11 to 55.79% ADF and from 0.96 to 9.94% ADL on a dry matter (DM) basis. With the use of the filter, the mean ADF values increased 4.2% and mean ADL values increased 18.9%. Overall, both ADF and ADL values were greater with the use of the glass microfiber filter than without, indicating that as the type of sample analyzed changed, use of the Gooch crucible without the filtering aid results in particle loss. The adoption of the use of a small pore size (1.5 μm) glass microfiber filter to improve filtration and recovery

  20. [Exposure to liquid detergent capsules: a study of the cases reported to the Paris Poison Center, 2011-2012].

    PubMed

    Villa, A; Médernach, C; Arropetian, N; Lagrange, F; Langrand, J; Garnier, R

    2014-06-01

    To evaluate the toxicity of liquid detergent capsules for children. Analysis of 684 consecutive cases from the Paris Poison Center (2011-2012). Most enquiries (97 %) concerned children 5 years of age or younger. The main circumstances of exposure were ingestion alone (72.4 %) or together with eye or skin contact (7.5 % and 7.3 %, respectively). The effects observed were generally due to the irritating properties of concentrated detergents: minor digestive disturbances (particularly vomiting in nearly 50 % of cases) after ingestion and conjunctivitis and/or keratitis after eye contact. The main complications were 24 cases of keratitis and one case of pulmonary toxicity after ingestion. A rash was observed in nine patients; it was delayed in two. The effects observed with liquid detergent capsules were very similar to those resulting from exposure to other detergents. However, exposure to these agents are very frequent and often results in eye contact, which may be responsible for keratitis, and after ingestion detergent inhalation is a possible complication. All cases with eye symptoms or cough after liquid detergent capsule exposure deserve prompt medical examination and assistance. Greater awareness of both health professionals and consumers on the dangers and risks of these laundry detergent pods is required for better treatment of exposure accidents and for their prevention. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  1. Improvement of the solubilization of proteins in two-dimensional electrophoresis with immobilized pH gradients

    PubMed Central

    Rabilloud, Thierry; Adessi, C.; Giraudel, A.; Lunardi, J.

    2007-01-01

    Summary We have carried out the separation of sparingly-soluble (membrane and nuclear) proteins by high resolution two-dimensional electrophoresis. IEF with immobilized pH gradients leads to severe quantitative losses of proteins in the resulting 2-D map, although the resolution is usually kept high. We therefore tried to improve the solubility of proteins in this technique, by using denaturing cocktails containing various detergents and chaotropes. Best results were obtained by using a denaturing solution containing urea, thiourea, and detergents (both nonionic and zwitterionic). The usefulness of thiourea-containing denaturing mixtures are shown in this article on several models including microsomal and nuclear proteins and on tubulin, a protein highly prone to aggregation. PMID:9150907

  2. Solubilization of active opiate receptors.

    PubMed Central

    Simonds, W F; Koski, G; Streaty, R A; Hjelmeland, L M; Klee, W A

    1980-01-01

    Receptors that reversibly bind opiates and opioid peptides have been solubilized from brain and neuroblastoma-glioma hybrid cell NG108-15 membranes. Active receptors are specifically solubilized with a new type of detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, which is a zwitterionic derivative of cholic acid. The solubilized receptor complexes behave as large molecules with a Stokes radius of 70 A and contain protein as an essential constituent. PMID:6254034

  3. A new high-performance thin layer chromatography-based assay of detergents and surfactants commonly used in membrane protein studies.

    PubMed

    Barret, Laurie-Anne; Polidori, Ange; Bonneté, Françoise; Bernard-Savary, Pierre; Jungas, Colette

    2013-03-15

    The hydrophobic nature of membrane proteins (MPs) necessitates the use of detergents for their extraction, solubilization and purification. Because the concentration of amphiphiles is crucial in the crystallization process, detergent quantification is essential to routine analysis. Here we describe a quantitative high-performance thin-layer chromatography (HPTLC) method we developed for the detection of small quantities of detergent bound to solubilized MPs. After optimization of aqueous deposit conditions, we show that most detergents widely used in membrane protein crystallography display distinctive mobilities in a mixture of dichloromethane, methanol and acetic acid 32:7.6:0.4 (v/v/v). Migration and derivatization conditions were optimized with n-dodecyl-β-D-maltoside (DDM), the most popular detergent for membrane protein crystallization. A linear calibration curve very well fits our data from 0.1 to 1.6 μg of DDM in water with a limit of detection of 0.05 μg. This limit of detection is the best achieved to date for a routine detergent assay, being not modified by the addition of NaCl, commonly used in protein buffers. With these chromatographic conditions, no prior treatment is required to assess the quantities of detergent bound to purified MPs, thus enabling the quantification of close structure detergents via a single procedure. This HPTLC method, which is fast and requires low sample volume, is fully suitable for routine measurements. Copyright © 2013 Elsevier B.V. All rights reserved.

  4. Thermostable, alkaline and detergent-tolerant lipase from a newly isolated thermophilic Bacillus stearothermophilus.

    PubMed

    Ben Bacha, Abir; Moubayed, Nadine M S; Abid, Islam

    2015-04-01

    Lipases are the enzymes of choice for laundry detergent industries, owing to their triglyceride removing ability from the soiled fabric, which eventually reduces the usage of phosphate-based chemical cleansers in the detergent formulation. In this study, a novel thermo-alkaline lipase-producing strain identified as Bacillus stearothermophilus was isolated from the soil samples of olive oil mill. Enhanced lipase production was observed at 55 degrees C, pH 11 and after 48 h of incubation. Among the substrates tested, xylose (a carbon source), peptone (a nitrogen source) and olive oil at a concentration of 1% were suitable substrates for enhancing lipase production. MgSO4 and Tween-80 were suitable substrates for maximizing lipase production. The enzyme was purified to homogeneity by a single CM-Sephadex column chromatography and revealed molecular mass of 67 kDa. The enzyme (BL1) was active over a wide range of pH from 9.0 to 13.0, with an optimum at pH 11.0, exhibited maximal activity at 55 degreesC and retained more than 70% of its activity after incubation at 70 degrees C or pH 13 for 0.5 h or 24 h, respectively. The enzyme hydrolyzed both short and long-chain triacylglycerols at comparable rates. BL1 was studied in a preliminary evaluation for use in detergent formulation solutions. This novel lipase showed extreme stability towards non-ionic and anionic surfactants after pre-incubation for 1 h at 40 degrees C, and good stability towards oxidizing agents. Additionally, the enzyme showed excellent stability and compatibility with various commercial detergents, suggesting its potential as an additive in detergent formulations.

  5. Enhanced gene delivery to the lung using biodegradable polyunsaturated cationic phosphatidylcholine-detergent conjugates.

    PubMed

    Pierrat, Philippe; Kereselidze, Dimitri; Lux, Marie; Lebeau, Luc; Pons, Françoise

    2016-09-10

    Lung diseases are among the more representative causes of mortality and morbidity worldwide and gene therapy is considered as a promising therapeutic approach for their treatment. However the design of efficient nucleic acid carriers for airway administration still is a challenge and there is a pressing need for new developments in this field. Herein, new synthetic DNA carriers based on the conjugation of a phospholipid and C12E4, a nonionic detergent, are developed. DNA complexes with phosphatidylcholine-detergent conjugates are administered in mouse airways, and transgene expression and inflammatory activity as an index of toxicity are investigated as a function of time, DNA dose, and presence of helper and stealth lipids. Introduction of a biodegradable linker between the phosphatidylcholine and detergent moieties significantly attenuates the severity of inflammatory response that characterizes cationic lipid-mediated gene transfer. Concurrent introduction of polyunsaturated fatty acid chains in the carrier scaffold improves transgene expression and further reduces airway inflammation. Finally, the biodegradable phosphatidylcholine-detergent conjugates favorably compare to GL67A, the gold standard for DNA delivery to the airway that is currently under clinical evaluation. Our findings indicate that the lipid formulations described herein may have great potential as nucleic acid carriers for gene therapy. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Differential detergent resistance of the apical and basolateral NPPases: relationship with polarized targeting.

    PubMed

    Delaunay, Jean-Louis; Breton, Michelyne; Goding, James W; Trugnan, Germain; Maurice, Michèle

    2007-03-15

    Targeting of glycosylphosphatidylinositol-anchored proteins to the apical surface of epithelial cells involves clustering in Triton X-100-resistant membrane microdomains or rafts. The role of these microdomains in sorting transmembrane proteins is more questionable because, unlike glycosylphosphatidylinositol-anchored proteins, apical transmembrane proteins are rather soluble in Triton X-100. They are, however, resistant to milder detergents such as Lubrol WX or Tween 20. It has been proposed that specific membrane microdomains, defined by resistance to these detergents, would carry transmembrane proteins to the apical surface. We have used MDCK cells stably transfected with the apical and basolateral pyrophosphatases/phosphodiesterases, NPP3 and NPP1, to examine the relationship between detergent resistance and apical targeting. The apically expressed wild-type NPP3 was insoluble in Lubrol WX whereas wild-type NPP1, which is expressed basolaterally, was essentially soluble. By using tail mutants and chimeric constructs that combine the cytoplasmic, transmembrane and extracellular domains of NPP1 and NPP3, we show that there is not a strict correlation between detergent resistance and apical targeting. Lubrol resistance is an intrinsic property of NPP3, which is acquired early during the biosynthetic process irrespective of its final destination, and depends on positively charged residues in its cytoplasmic tail.

  7. Wheat germ cell-free expression: Two detergents with a low critical micelle concentration allow for production of soluble HCV membrane proteins.

    PubMed

    Fogeron, Marie-Laure; Badillo, Aurélie; Jirasko, Vlastimil; Gouttenoire, Jérôme; Paul, David; Lancien, Loick; Moradpour, Darius; Bartenschlager, Ralf; Meier, Beat H; Penin, François; Böckmann, Anja

    2015-01-01

    Membrane proteins are notoriously difficult to express in a soluble form. Here, we use wheat germ cell-free expression in the presence of various detergents to produce the non-structural membrane proteins 2, 4B and 5A of the hepatitis C virus (HCV). We show that lauryl maltose neopentyl glycol (MNG-3) and dodecyl octaethylene glycol ether (C12E8) detergents can yield essentially soluble membrane proteins at detergent concentrations that do not inhibit the cell-free reaction. This finding can be explained by the low critical micelle concentration (CMC) of these detergents, which keeps the monomer concentrations low while at the same time providing the necessary excess of detergent concentration above CMC required for full target protein solubilization. We estimate that a tenfold excess of detergent micelles with respect to the protein concentration is sufficient for solubilization, a number that we propose as a guideline for detergent screening assays. Copyright © 2014 Elsevier Inc. All rights reserved.

  8. Diester-containing Zwitterionic Gemini Surfactants with Different Spacer and Its Impact on Micellization Properties and Viscosity of Aqueous Micellar Solution.

    PubMed

    Patil, Sachin Vasant; Patil, Sanyukta Arun; Pratap, Amit Prabhakar

    2016-09-01

    A series of diester containing zwitterionic gemini surfactants, N,N-dimethyl-N-alkyl-2-[[hydroxy (alkoxy) phosphinyl]oxy]-alkylammonium designated as C8(-)-S-Cn(+), S = 2 and 3, n = 12, 14 and 16, were synthesized and characterized by instrumental techniques namely FT-IR, (1)H NMR, (13)C NMR, (31)P NMR and Mass spectral studies. These new gemini surfactants further investigated for their various surfactant properties. The critical micelle concentration (cmc) and the effectiveness of surface tension reduction (Πcmc) were determined as a function of surfactant concentration by means of surface tension measurement. Micellization and viscosity properties were investigated by surface tension, electrical conductivity, dye micellization and rheology techniques. The findings of the aqueous surfactant system obtained were impacted by polarity, size and the nature of zwitterions as the surface. The thermodynamic and viscosity properties of these surfactants found to be based on the structures of gemini surfactants.

  9. Removal of detergents from protein extracts using activated charcoal prior to immunological analysis.

    PubMed

    Malhas, Ashraf N; Abuknesha, Ramadan A; Price, Robert G

    2002-06-01

    The use of dextran-coated activated charcoal (DCC) powder to absorb solubilising detergents from cell lysates is described. Normal embryonic epithelial cells were lysed in the presence of sodium dodecyl sulphate (SDS). The detergent was then absorbed with DCC to facilitate analysis of polycystin-1 with antibody-based methods. Polycystin-1 is a membrane protein that is involved in the pathogenesis of autosomal dominant polycystic kidney disease (ADPKD). The adverse effect of SDS on antibody-polycystin-1 binding was studied and the improvement resulting from its removal demonstrated using enzyme-linked immunosorbent assays (ELISAs). The results indicate that DCC can be used in a simple manner to remove highly reactive membrane-solubilising reagents from protein mixtures prior to immunological analysis. This procedure may be relevant to a variety of other techniques that are normally affected by detergents.

  10. Design, Synthesis, and Characterization of Novel Zwitterionic Lipids for Drug and siRNA Delivery Applications

    NASA Astrophysics Data System (ADS)

    Walsh, Colin L.

    Lipid-based nanoparticles have long been used to deliver biologically active molecules such as drugs, proteins, peptides, DNA, and siRNA in vivo. Liposomes and lipoplexes alter the biodistribution, pharmacokinetics, and cellular uptake of their encapsulated or associated cargo. This can increase drug efficacy while reducing toxicity, resulting in an increased therapeutic index and better clinical outcomes. Unlike small molecule drugs, which passively diffuse through lipid membranes, nucleic acids and proteins require an active, carrier mediated escape mechanism to reach their site of action. As such, the therapeutic application and drug properties dictate the required biophysical characteristics of the lipid nanoparticle. These carrier properties depend on the structure and biophysical characteristics of the lipids and other components used to formulate them. This dissertation presents a series of studies related to the development of novel synthetic lipids for use in drug delivery systems. First, we developed a novel class of zwitterionic lipids with head groups containing a cationic amine and anionic carboxylate and ester-linked oleic acid tails. These lipids exhibit structure-dependent, pH-responsive biophysical properties, and may be useful components for next-generation drug delivery systems. Second, we extended the idea of amine/carboxylate containing zwitterionic head groups and synthesized a series of acetate terminated diacyl lipids containing a quaternary amine. These lipids have an inverted headgroup orientation compared to naturally occurring zwitterionic lipids, and show interesting salt-dependent biophysical properties. Third, we synthesized and characterized a focused library of ionizable lysine-based lipids, which contain a lysine head group linked to a long-chain dialkylamine. A focused library was synthesized to determine the impact of hydrophobic fluidity, lipid net charge, and lipid pKa on the biophysical and siRNA transfection characteristics

  11. Applying thermosettable zwitterionic copolymers as general fouling-resistant and thermal-tolerant biomaterial interfaces.

    PubMed

    Chou, Ying-Nien; Chang, Yung; Wen, Ten-Chin

    2015-05-20

    We introduced a thermosettable zwitterionic copolymer to design a high temperature tolerance biomaterial as a general antifouling polymer interface. The original synthetic fouling-resistant copolymer, poly(vinylpyrrolidone)-co-poly(sulfobetaine methacrylate) (poly(VP-co-SBMA)), is both thermal-tolerant and fouling-resistant, and the antifouling stability of copolymer coated interfaces can be effectively controlled by regulating the VP/SBMA composition ratio. We studied poly(VP-co-SBMA) copolymer gels and networks with a focus on their general resistance to protein, cell, and bacterial bioadhesion, as influenced by the thermosetting process. Interestingly, we found that the shape of the poly(VP-co-SBMA) copolymer material can be set at a high annealing temperature of 200 °C while maintaining good antifouling properties. However, while the zwitterionic PSBMA polymer gels were bioinert as expected, control of the fouling resistance of the PSBMA polymer networks was lost in the high temperature annealing process. A poly(VP-co-SBMA) copolymer network composed of PSBMA segments at 32 mol % showed reduced fibrinogen adsorption, tissue cell adhesion, and bacterial attachment, but a relatively higher PSBMA content of 61 mol % was required to optimize resistance to platelet adhesion and erythrocyte attachment to confer hemocompatibility to human blood. We suggest that poly(VP-co-SBMA) copolymers capable of retaining stable fouling resistance after high temperature shaping have a potential application as thermosettable materials in a bioinert interface for medical devices, such as the thermosettable coating on a stainless steel blood-compatible metal stent investigated in this study.

  12. Detergents enhance EspB secretion from Escherichia coli strains harboring the locus for the enterocyte effacement (LEE) gene.

    PubMed

    Nakasone, Noboru; Toma, Claudia; Higa, Naomi; Koizumi, Yukiko; Ogura, Yasunori; Suzuki, Toshihiko

    2011-02-01

    The effects of detergents (cholic acid, deoxycholic acid, Triton X-100, and Nonidet P-40) on the secretion of EspB from the locus for enterocyte effacement (LEE) gene-positive Escherichia coli strains were examined. Clinical isolates of eight EPEC strains and seven STEC strains were used to detect EspB after they had been cultivated in Luria-Bertani (LB) broth containing one of the detergents. When the bacteria were cultured in LB broth supplemented with one of the detergents, the amount of EspB produced was increased by 2-32-fold depending on the detergent and the strain used. EspB was detected in all strains when they were cultured in LB broth containing all of the detergents. The results obtained in this study can be applied to immunological diagnostic methods for detecting EspB and also to the production of EspB for research purposes. © 2010 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  13. Nonionic and zwitterionic forms of glycylglycylarginine as a part of spider silk protein: Spectroscopic and theoretical study

    NASA Astrophysics Data System (ADS)

    Arı, Hatice; Özpozan, Talat

    2016-01-01

    Glycylglycylarginine as a part of GGX motif of spider silk spidroin in nonionic (non-GGR) and zwitterionic (zwt-GGR) forms have been examined from theoretical and spectroscopic aspects. The most stable conformational isomers of non-GGR and zwt-GGR were obtained through relaxed scan using the DFT/B3LYP with 6-31G(d) basis set. Nonionic and zwitterionic forms of 310-helix structures of GGR have also been calculated and compared with the most stable conformers obtained as a result of conformer analysis of isolated three peptide structures. This comparison should give an idea about the stability contribution of intermolecular interactions between the 310-helix structured peptide chains. O3LYP and B3PW91 hybrid functionals beside B3LYP have also been used for further calculations of geometry optimization, vibrational analysis, Natural Bond Orbital (NBO) analysis, HOMO-LUMO analysis and hydrogen bonding analysis. Normal Mode Analysis was carried through Potential Energy Distribution (PED) calculations by means of VEDA4 program package. IR and Raman spectra of GGR have also been used to relate the spectroscopic data obtained to electronic and structural features.

  14. Nonionic and zwitterionic forms of glycylglycylarginine as a part of spider silk protein: Spectroscopic and theoretical study.

    PubMed

    Arı, Hatice; Özpozan, Talat

    2016-01-05

    Glycylglycylarginine as a part of GGX motif of spider silk spidroin in nonionic (non-GGR) and zwitterionic (zwt-GGR) forms have been examined from theoretical and spectroscopic aspects. The most stable conformational isomers of non-GGR and zwt-GGR were obtained through relaxed scan using the DFT/B3LYP with 6-31G(d) basis set. Nonionic and zwitterionic forms of 310-helix structures of GGR have also been calculated and compared with the most stable conformers obtained as a result of conformer analysis of isolated three peptide structures. This comparison should give an idea about the stability contribution of intermolecular interactions between the 310-helix structured peptide chains. O3LYP and B3PW91 hybrid functionals beside B3LYP have also been used for further calculations of geometry optimization, vibrational analysis, Natural Bond Orbital (NBO) analysis, HOMO-LUMO analysis and hydrogen bonding analysis. Normal Mode Analysis was carried through Potential Energy Distribution (PED) calculations by means of VEDA4 program package. IR and Raman spectra of GGR have also been used to relate the spectroscopic data obtained to electronic and structural features. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Structural, quantum chemical, vibrational and thermal studies of a hydrogen bonded zwitterionic co-crystal (nicotinic acid: pyrogallol)

    NASA Astrophysics Data System (ADS)

    Prabha, E. Arockia Jeya Yasmi; Kumar, S. Suresh; Athimoolam, S.; Sridhar, B.

    2017-02-01

    In the present work, a new co-crystal of nicotinic acid with pyrogallol (NICPY) has been grown in the zwitterionic form and the corresponding structural, vibrational, thermal, solubility and anti-cancer characteristics have been reported. The single crystal X-ray diffraction analysis confirms that the structural molecular packing of the crystal stabilized through N-H⋯O and O-H⋯O hydrogen bond. The stabilization energy of the hydrogen bond motifs were calculated in the solid state. Vibrational spectral studies such as Fourier transform-infrared (FT-IR) and FT-Raman were adopted to understand the zwitterionic co-crystalline nature of the compound, which has been compared with theoretically calculated vibrational frequencies. The thermal stability of the grown co-crystal was analyzed by TG/DTA study. The solubility of the NICPY co-crystal was investigated in water at different temperature and compared with that of the nicotinic acid, which is the parent compound of NICPY co-crystal. The grown crystals were treated with human cervical cancer cell line (HeLa) to analyze the cytotoxicity of NICPY crystals and compared with the parent compound, which shows that NICPY has moderate activity against human cervical cancer cell line.

  16. Photodetachment of Zwitterions: Probing Intramolecular Coulomb Repulsion and Attraction in the Gas Phase Using Mono- Decarboxylated Pyridinium Dicarboxylates. Implications on the Mechanism of Orotidine 5' -Monophosphate Decarboxylase

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

    Wang, Xue B.; Dacres, J E.; Yang, Xin

    2003-11-11

    Negative ion photoelectron spectra resulting from the decarboxylation of nine zwitterionic pyridinium dicarboxylates (D(x,y)) are reported. Structural assignments are made on the basis of analogy to the spectra of related species, labeling experiments with C-13- or H-2-containing substrates, independent syntheses, and comparison to density functional theory and ab initio (B3LYP and CCSD(T), respectively) results. In some cases, an acid-catalyzed isomerization of the D(xy)-CO2 ions was found to take place. Adiabatic detachment energies of the resulting zwitterionic ions were measured and are well reproduced by theory. The relative stabilities of the D(x,Y)- CO2 decarboxylation products are largely determined by their intramolecularmore » electrostatic interactions, which are directly probed by the photoelectron spectra and were analyzed in terms of the resulting Coulombic forces. Expulsion of carbon dioxide from the D(x,y) ions was also used as an electrostatic model to probe the mechanism of the enzyme-catalyzed conversion of orotidine 5'-monophosphate (OMP) to uridine 5'-monophosphate (UMP). It was found that the loss of CO2 from these zwitterions; and from oxygen-protonated OMP is retarded by the presence of an additional anionic group. This suggests that the formation of a zwitterion intermediate in the enzyme-catalyzed transformation of OMP to UMP may have less of an energetic impact than commonly thought and could be a''red herring''. If so, the electrostatic stress mechanism proposed by Larsen et al. and Pai, Guo, and co-workers maybe followed.« less

  17. Development of USEtox characterisation factors for dishwasher detergents using data made available under REACH.

    PubMed

    Igos, Elorri; Moeller, Ruth; Benetto, Enrico; Biwer, Arno; Guiton, Mélanie; Dieumegard, Philippe

    2014-04-01

    Because of the more and more stringent regulations and customer demand, dishwasher detergent manufacturers are constantly improving the composition of the products towards better environmental performances. In order to quantify the pros and cons of these changes on the lifecycle of detergents, as compared to conventional products, the use of Life Cycle Assessment (LCA) is a meaningful opportunity. However, the application of the methodology is hampered by the lack of Characterisation Factors (CFs) relative to the specific chemical substances included in the detergents composition, which cannot be included in the impact assessment of the effluent discharge. In this study we have tackled this problem, taking advantage of the specific case of three dishwasher detergents produced by the Chemolux/McBride group: phosphate-based, eco-labelled and phosphate-free formulations. Nine CFs for freshwater ecotoxicity and seven CFs for human toxicity have been developed, using the USEtox methodology and data made available under the REACH regulation. As a result, the dishwasher effluent composition could be characterised by more than 95% for freshwater ecotoxicity whereas for human toxicity the percentage was less than 36%, due to the lack of adequate and reliable repeated dose toxicity studies. The main contributing substances to freshwater ecotoxicity were found to be sodium percarbonate and sodium triphosphate, the latter confirming the pertinence of phosphates banning in the detergent industry. Regarding human toxicity, zinc shows the highest contribution. Further comparison to previous studies and sensitivity analysis substantiated the robustness of these conclusions. Copyright © 2013 Elsevier Ltd. All rights reserved.

  18. Characterization of detergent-solubilized sarcoplasmic reticulum Ca2+-ATPase by high-performance liquid chromatography.

    PubMed

    Andersen, J P; Vilsen, B; Nielsen, H; Møller, J V

    1986-10-21

    Sarcoplasmic reticulum Ca2+-ATPase solubilized by the nonionic detergent octaethylene glycol monododecyl ether was studied by molecular sieve high-performance liquid chromatography (HPLC) and analytical ultracentrifugation. Significant irreversible aggregation of soluble Ca2+-ATPase occurred within a few hours in the presence of less than or equal to 50 microM Ca2+. The aggregates were inactive and were primarily held together by hydrophobic forces. In the absence of reducing agent, secondary formation of disulfide bonds occurred. The stability of the inactive dimer upon dilution permitted unambiguous assignment of its elution position and sedimentation coefficient. At high Ca2+ concentration (500 microM), monomeric Ca2+-ATPase was stable for several hours. Reversible self-association induced by variation in protein, detergent, and lipid concentrations was studied by large-zone HPLC. The association constant for dimerization of active Ca2+-ATPase was found to be 10(5)-10(6) M-1 depending on the detergent concentration. More detergent was bound to monomeric than to dimeric Ca2+-ATPase, even above the critical micellar concentration of the detergent. Binding of Ca2+ and vanadate as well as ATP-dependent phosphorylation was studied in monomeric and in reversibly associated dimeric preparations. In both forms, two high-affinity Ca2+ binding sites per phosphorylation site existed. The delipidated monomer purified by HPLC was able to form ADP-insensitive phosphoenzyme and to bind ATP and vanadate simultaneously. These results suggest that formation of Ca2+-ATPase oligomers in the membrane is governed by nonspecific forces (low affinity) and that each polypeptide chain constitutes a functional unit.

  19. Characterization of Hydrophobic Peptides in the Presence of Detergent by Photoionization Mass Spectrometry

    PubMed Central

    Bagag, Aïcha; Jault, Jean-Michel; Sidahmed-Adrar, Nazha; Réfrégiers, Matthieu; Giuliani, Alexandre; Le Naour, François

    2013-01-01

    The characterization of membrane proteins is still challenging. The major issue is the high hydrophobicity of membrane proteins that necessitates the use of detergents for their extraction and solubilization. The very poor compatibility of mass spectrometry with detergents remains a tremendous obstacle in studies of membrane proteins. Here, we investigated the potential of atmospheric pressure photoionization (APPI) for mass spectrometry study of membrane proteins. This work was focused on the tetraspanin CD9 and the multidrug transporter BmrA. A set of peptides from CD9, exhibiting a broad range of hydropathicity, was investigated using APPI as compared to electrospray ionization (ESI). Mass spectrometry experiments revealed that the most hydrophobic peptides were hardly ionized by ESI whereas all peptides, including the highly hydrophobic one that corresponds to the full sequence of the first transmembrane domain of CD9, were easily ionized by APPI. The native protein BmrA purified in the presence of the non-ionic detergent beta-D-dodecyl maltoside (DDM) was digested in-solution using trypsin. The resulting peptides were investigated by flow injection analysis of the mixture followed by mass spectrometry. Upon ESI, only detergent ions were detected and the ionic signals from the peptides were totally suppressed. In contrast, APPI allowed many peptides distributed along the sequence of the protein to be detected. Furthermore, the parent ion corresponding to the first transmembrane domain of the protein BmrA was detected under APPI conditions. Careful examination of the APPI mass spectrum revealed a-, b-, c- and y- fragment ions generated by in-source fragmentation. Those fragment ions allowed unambiguous structural characterization of the transmembrane domain. In conclusion, APPI–MS appears as a versatile method allowing the ionization and fragmentation of hydrophobic peptides in the presence of detergent. PMID:24236085

  20. 40 CFR 80.161 - Detergent additive certification program.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... corresponding test results (percent flow restriction demonstrated in the PFID test and milligrams of deposit per... penalties in §§ 80.169 and 80.172. (iii) If both parties submit the required information, EPA will evaluate... customers who use the disqualified detergent. Failure to do so may subject the certifier to liabilities for...

  1. Screening of detergents for solubilization, purification and crystallization of membrane proteins: a case study on succinate:ubiquinone oxidoreductase from Escherichia coli.

    PubMed

    Shimizu, Hironari; Nihei, Coh-ichi; Inaoka, Daniel Ken; Mogi, Tatushi; Kita, Kiyoshi; Harada, Shigeharu

    2008-09-01

    Succinate:ubiquinone oxidoreductase (SQR) was solubilized and purified from Escherichia coli inner membranes using several different detergents. The number of phospholipid molecules bound to the SQR molecule varied greatly depending on the detergent combination that was used for the solubilization and purification. Crystallization conditions were screened for SQR that had been solubilized and purified using 2.5%(w/v) sucrose monolaurate and 0.5%(w/v) Lubrol PX, respectively, and two different crystal forms were obtained in the presence of detergent mixtures composed of n-alkyl-oligoethylene glycol monoether and n-alkyl-maltoside. Crystallization took place before detergent phase separation occurred and the type of detergent mixture affected the crystal form.

  2. A newly high alkaline lipase: an ideal choice for application in detergent formulations

    PubMed Central

    2011-01-01

    Background Bacterial lipases received much attention for their substrate specificity and their ability to function in extreme environments (pH, temperature...). Many staphylococci produced lipases which were released into the culture medium. Reports of thermostable lipases from Staphylococcus sp. and active in alkaline conditions are not previously described. Results A newly soil-isolated Staphylococcus sp. strain ESW secretes an induced lipase in the culture medium. The effects of temperature, pH and various components in a detergent on the activity and stability of Staphylococcus sp. lipase (SL1) were studied in a preliminary evaluation for use in detergent formulation solutions. The enzyme was highly active over a wide range of pH from 9.0 to 13.0, with an optimum at pH 12.0. The relative activity at pH 13.0 was about 60% of that obtained at pH 12.0. It exhibited maximal activity at 60°C. This novel lipase, showed extreme stability towards non-ionic and anionic surfactants after pre-incubation for 1 h at 40°C, and relative stability towards oxidizing agents. Additionally, the crude enzyme showed excellent stability and compatibility with various commercial solid and liquid detergents. Conclusions These properties added to the high activity in high alkaline pH make this novel lipase an ideal choice for application in detergent formulations. PMID:22123072

  3. Quantitative Determination of NTA and Other Chelating Agents in Detergents by Potentiometric Titration with Copper Ion Selective Electrode.

    PubMed

    Ito, Sana; Morita, Masaki

    2016-01-01

    Quantitative analysis of nitrilotriacetate (NTA) in detergents by titration with Cu 2+ solution using a copper ion selective electrode was achieved. This method tolerates a wide range of pH and ingredients in detergents. In addition to NTA, other chelating agents, having relatively lower stability constants toward Cu 2+ , were also qualified with sufficient accuracy by this analytical method for model detergent formulations. The titration process was automated by automatic titrating systems available commercially.

  4. Isolation of a Moderately Stable but Sensitive Zwitterionic Diazonium Tetrazolyl-1,2,3-triazolate.

    PubMed

    Klapötke, Thomas M; Krumm, Burkhard; Pflüger, Carolin

    2016-07-15

    An unexpected formation of a diazonium compound was observed by nitration of an amino substituted triazolyl tetrazole with mixed acid. The crystal structure determination revealed a zwitterionic diazonium tetrazolyl-1,2,3-triazolate, whose constitution was supported by NMR and vibrational spectroscopic analysis. The thermal stability and sensitivity toward impact and friction were determined. In contrast, diazotriazoles are rather unstable and are mainly handled in solution and/or low temperatures, which is not the case for this diazonium tetrazolyl-1,2,3-triazolate, being stable at ambient temperature.

  5. Outcomes Related to the Use of Frozen Plasma or Pooled Solvent/Detergent-Treated Plasma in Critically Ill Children.

    PubMed

    Camazine, Maraya N; Karam, Oliver; Colvin, Ryan; Leteurtre, Stephane; Demaret, Pierre; Tucci, Marisa; Muszynski, Jennifer A; Stanworth, Simon; Spinella, Philip C

    2017-05-01

    To determine if the use of fresh frozen plasma/frozen plasma 24 hours compared to solvent detergent plasma is associated with international normalized ratio reduction or ICU mortality in critically ill children. This is an a priori secondary analysis of a prospective, observational study. Study groups were defined as those transfused with either fresh frozen plasma/frozen plasma 24 hours or solvent detergent plasma. Outcomes were international normalized ratio reduction and ICU mortality. Multivariable logistic regression was used to determine independent associations. One hundred one PICUs in 21 countries. All critically ill children admitted to a participating unit were included if they received at least one plasma unit during six predefined 1-week (Monday to Friday) periods. All children were exclusively transfused with either fresh frozen plasma/frozen plasma 24 hours or solvent detergent plasma. None. There were 443 patients enrolled in the study. Twenty-four patients (5%) were excluded because no plasma type was recorded; the remaining 419 patients were analyzed. Fresh frozen plasma/frozen plasma 24 hours group included 357 patients, and the solvent detergent plasma group included 62 patients. The median (interquartile range) age and weight were 1 year (0.2-6.4) and 9.4 kg (4.0-21.1), respectively. There was no difference in reason for admission, severity of illness score, pretransfusion international normalized ratio, or lactate values; however, there was a difference in primary indication for plasma transfusion (p < 0.001). There was no difference in median (interquartile range) international normalized ratio reduction, between fresh frozen plasma/frozen plasma 24 hours and solvent detergent plasma study groups, -0.2 (-0.4 to 0) and -0.2 (-0.3 to 0), respectively (p = 0.80). ICU mortality was lower in the solvent detergent plasma versus fresh frozen plasma/frozen plasma 24 hours groups, 14.5% versus 29.1%%, respectively (p = 0.02). Upon adjusted analysis

  6. Screening of detergents for solubilization, purification and crystallization of membrane proteins: a case study on succinate:ubiquinone oxidoreductase from Escherichia coli

    PubMed Central

    Shimizu, Hironari; Nihei, Coh-ichi; Inaoka, Daniel Ken; Mogi, Tatushi; Kita, Kiyoshi; Harada, Shigeharu

    2008-01-01

    Succinate:ubiquinone oxidoreductase (SQR) was solubilized and purified from Escherichia coli inner membranes using several different detergents. The number of phospholipid molecules bound to the SQR molecule varied greatly depending on the detergent combination that was used for the solubilization and purification. Crystallization conditions were screened for SQR that had been solubilized and purified using 2.5%(w/v) sucrose monolaurate and 0.5%(w/v) Lubrol PX, respectively, and two different crystal forms were obtained in the presence of detergent mixtures composed of n-alkyl-oligoethylene glycol monoether and n-alkyl-maltoside. Crystallization took place before detergent phase separation occurred and the type of detergent mixture affected the crystal form. PMID:18765923

  7. Conformational thermostabilization of the β1-adrenergic receptor in a detergent-resistant form

    PubMed Central

    Serrano-Vega, Maria J.; Magnani, Francesca; Shibata, Yoko; Tate, Christopher G.

    2008-01-01

    There are ≈350 non-odorant G protein-coupled receptors (GPCRs) encoded by the human genome, many of which are predicted to be potential therapeutic targets, but there are only two structures available to represent the whole of the family. We hypothesized that improving the detergent stability of these receptors and simultaneously locking them into one preferred conformation will greatly improve the chances of crystallization. We developed a generic strategy for the isolation of detergent-solubilized thermostable mutants of a GPCR, the β1-adrenergic receptor. The most stable mutant receptor, βAR-m23, contained six point mutations that led to an apparent Tm 21°C higher than the native protein, and, in the presence of bound antagonist, βAR-m23 was as stable as bovine rhodopsin. In addition, βAR-m23 was significantly more stable in a wide range of detergents ideal for crystallization and was preferentially in an antagonist conformation in the absence of ligand. PMID:18192400

  8. Bile salts stimulate mucin secretion by cultured dog gallbladder epithelial cells independent of their detergent effect.

    PubMed

    Klinkspoor, J H; Yoshida, T; Lee, S P

    1998-05-15

    1. Bile salts stimulate mucin secretion by the gallbladder epithelium. We have investigated whether this stimulatory effect is due to a detergent effect of bile salts. 2. The bile salts taurocholic acid (TC) and tauroursodeoxycholic acid (TUDC) and the detergents Triton X-100 (12.5-400 microM) and Tween-20 (0.1-3.2 mM) were applied to monolayers of cultured dog gallbladder epithelial cells. Mucin secretion was studied by measuring the secretion of [3H]N-acetyl-d-glucosamine-labelled glycoproteins. We also attempted to alter the fluidity of the apical membrane of the cells through extraction of cholesterol with beta-cyclodextrin (2.5-15 mM). The effect on TUDC-induced mucin secretion was studied. Cell viability was assessed by measuring lactate dehydrogenase (LDH) leakage or 51Cr release. 3. In contrast with the bile salts, the detergents were not able to cause an increase in mucin secretion without causing concomitant cell lysis. Concentrations of detergent that increased mucin release (>100 microM Triton X-100, >0.8 mM Tween-20), caused increased LDH release. Incubation with beta-cyclodextrin resulted in effective extraction of cholesterol without causing an increase in 51Cr release. However, no effect of the presumed altered membrane fluidity on TUDC (10 mM)-induced mucin secretion was observed. 4. The stimulatory effect of bile salts on mucin secretion by gallbladder epithelial cells is not affected by the fluidity of the apical membrane of the cells and also cannot be mimicked by other detergents. We conclude that the ability of bile salts to cause mucin secretion by the gallbladder epithelium is not determined by their detergent properties.

  9. Solution structure of detergent micelles at conditions relevant to membrane protein crystallization.

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

    Littrell, K.; Thiyagarajan, P.; Tiede, D.

    1999-07-02

    In this study small angle neutron scattering was used to characterize the formation of micelles in aqueous solutions of the detergents DMG and SPC as a function of detergent concentration and ionic strength of the solvent. The effects on the micelle structure of the additives glycerol and PEG, alone as well as in combination typical for actual membrane protein crystallization, were also explored. This research suggests that the micelles are cigar-like in form at the concentrations studied. The size of the micelles was observed to increase with increasing ionic strength but decrease with the addition of glycerol or PEG.

  10. Maximizing detergent stability and functional expression of a GPCR by exhaustive recombination and evolution.

    PubMed

    Schlinkmann, Karola M; Hillenbrand, Matthias; Rittner, Alexander; Künz, Madeleine; Strohner, Ralf; Plückthun, Andreas

    2012-09-21

    To identify structural features in a G-protein-coupled receptor (GPCR) crucial for biosynthesis, stability in the membrane and stability in detergent micelles, we developed an evolutionary approach using expression in the inner membrane of Escherichia coli. From the analysis of 800,000 sequences of the rat neurotensin receptor 1, in which every amino acid had been varied to all 64 codons, we uncovered several "shift" positions, where the selected population focuses on a residue different from wild type. Here, we employed in vitro DNA recombination and a comprehensive synthetic binary library made by the Slonomics® technology, allowing us to uncover additive and synergistic effects in the structure that maximize both detergent stability and functional expression. We identified variants with >25,000 functional molecules per E. coli cell, a 50-fold increase over wild type, and observed strong coevolution of detergent stability. We arrived at receptor variants highly stable in short-chain detergents, much more so than those found by alanine scanning on the same receptor. These evolved GPCRs continue to be able to signal through the G-protein. We discuss the structural reasons for these improvements achieved through directed evolution. Copyright © 2012 Elsevier Ltd. All rights reserved.

  11. Preparation and characterization of novel zwitterionic poly(arylene ether sulfone) ultrafiltration membrane with good thermostability and excellent antifouling properties

    NASA Astrophysics Data System (ADS)

    Rong, Guolong; Zhou, Di; Han, Xiaocui; Pang, Jinhui

    2018-01-01

    Zwitterionic poly(arylene ether sulfone) (PAES-NS) was synthesized via copolymerization by using a bisphenol monomer with a pyridine group. The chemical structures of the copolymers were confirmed by using Fourier transform infrared (FTIR) and 1H nuclear magnetic resonance (NMR) spectroscopy; the copolymers showed good thermal stability. A series of polyphenysulfone (PPSU)/PAES-NS blend ultrafiltration (UF) membranes was prepared via conventional immersion precipitation phase inversion methods The morphologies of the modified membranes were investigated by scanning electron microscopy (SEM). The surface hydrophilicity of the UF membranes was studied by water contact angle measurement, indicating that the zwitterionic group increased the membrane hydrophilicity. UF of solvated model pollutants using the membranes showed a significant reduction of the irreversible adsorption of the foulants, illustrating the excellent anti-fouling properties of the membrane. The water flux of the PAES-NS membrane was significantly enhanced, being almost three times higher than that of the pristine PPSU membrane, with retention of a high rejection level. After three UF cycles, the water flux recovery of the PAES-NS membrane was as high as 96%.

  12. Impact of wash cycle time, temperature and detergent formulation on the hygiene effectiveness of domestic laundering.

    PubMed

    Honisch, M; Stamminger, R; Bockmühl, D P

    2014-12-01

    Investigation of the effect of temperature and duration of the laundering process with and without activated oxygen bleach (AOB)-containing detergent on the hygienic effectiveness of laundering. Cotton test swatches were contaminated with Staphylococcus aureus, Enterococcus hirae, Pseudomonas aeruginosa, Candida albicans and Trichophyton mentagrophytes and were washed in a household washing machine using temperatures between 20 and 60°C and different wash cycle times. The logarithmic microbial reduction factor and cross-contamination (i.e. transfer from contaminated to sterile swatches) were used to indicate the hygienic effectiveness of the washing process. For all tested micro-organisms, the temperature needed for decontamination depended on washing time and detergent type. Hygiene effectiveness of laundering was enhanced by inclusion of AOB even at lowest temperatures, except for C. albicans, which was virtually unaffected by AOB. The use of AOB-containing detergents as well as high washing temperatures reduced cross-contamination to sterile swatches included in the load. Depending on the type of organism, longer wash cycle times or the use of AOB-containing detergents can be used to enhance the hygiene effectiveness of laundering. The study demonstrates that it is possible to compensate for the loss of hygiene effectiveness of laundering at lower temperatures using detergents with activated oxygen bleach or by extending the wash cycle time. © 2014 The Society for Applied Microbiology.

  13. Stability of thermostable alkaline protease from Bacillus licheniformis RP1 in commercial solid laundry detergent formulations.

    PubMed

    Sellami-Kamoun, Alya; Haddar, Anissa; Ali, Nedra El-Hadj; Ghorbel-Frikha, Basma; Kanoun, Safia; Nasri, Moncef

    2008-01-01

    The stability of crude extracellular protease produced by Bacillus licheniformis RP1, isolated from polluted water, in various solid laundry detergents was investigated. The enzyme had an optimum pH and temperature at pH 10.0-11.0 and 65-70 degrees C. Enzyme activity was inhibited by PMSF, suggesting that the preparation contains a serine-protease. The alkaline protease showed extreme stability towards non-ionic (5% Tween 20% and 5% Triton X-100) and anionic (0.5% SDS) surfactants, which retained 100% and above 73%, respectively, of its initial activity after preincubation 60 min at 40 degrees C. The RP1 protease showed excellent stability and compatibility with a wide range of commercial solid detergents at temperatures from 40 to 50 degrees C, suggesting its further application in detergent industry. The enzyme retained 95% of its initial activity with Ariel followed by Axion (94%) then Dixan (93.5%) after preincubation 60 min at 40 degrees C in the presence of 7 mg/ml of detergents. In the presence of Nadhif and New Det, the enzyme retained about 83.5% of the original activity. The effects of additives such as maltodextrin, sucrose and PEG 4000 on the stability of the enzyme during spray-drying and during subsequent storage in New Det detergent were also examined. All additives tested enhanced stability of the enzyme.

  14. An impedimetric chemical sensor for determination of detergents residues.

    PubMed

    Bratov, Andrey; Abramova, Natalia; Ipatov, Andrey; Merlos, Angel

    2013-03-15

    A new impedimetric sensor based on an interdigitated electrode array with electrode digits located at the bottom of microcapillaries formed in silicon dioxide is presented. Microcapillaries are opened at the top, so that in contact with an electrolyte solution the ac current flows close to the surface of the capillary wall from one electrode to another and is significantly affected by changes in the surface conductance at the SiO2/electrolyte interface. Adsorption of detergents on the sensor surface affects the charge distribution in the electrical double layer and thus the surface conductance. These changes are registered by measuring impedance. Effect of surface adsorption of ionic and non-ionic surfactants on the sensor impedance is studied. The sensor is shown to be able to measure commercial detergents residues in a tap water starting from 5 ppm even in solutions with high electrolyte conductivity. Copyright © 2012 Elsevier B.V. All rights reserved.

  15. 21 CFR 701.20 - Detergent substances, other than soap, intended for use in cleansing the body.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 7 2013-04-01 2013-04-01 false Detergent substances, other than soap, intended... Ingredients § 701.20 Detergent substances, other than soap, intended for use in cleansing the body. (a) In its definition of the term cosmetic, the Federal Food, Drug, and Cosmetic Act specifically excludes soap. The...

  16. 21 CFR 701.20 - Detergent substances, other than soap, intended for use in cleansing the body.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 7 2012-04-01 2012-04-01 false Detergent substances, other than soap, intended... Ingredients § 701.20 Detergent substances, other than soap, intended for use in cleansing the body. (a) In its definition of the term cosmetic, the Federal Food, Drug, and Cosmetic Act specifically excludes soap. The...

  17. 21 CFR 701.20 - Detergent substances, other than soap, intended for use in cleansing the body.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 7 2014-04-01 2014-04-01 false Detergent substances, other than soap, intended... Ingredients § 701.20 Detergent substances, other than soap, intended for use in cleansing the body. (a) In its definition of the term cosmetic, the Federal Food, Drug, and Cosmetic Act specifically excludes soap. The...

  18. Versatile antifouling polyethersulfone filtration membranes modified via surface grafting of zwitterionic polymers from a reactive amphiphilic copolymer additive.

    PubMed

    Zhao, Yi-Fan; Zhang, Pei-Bin; Sun, Jian; Liu, Cui-Jing; Yi, Zhuan; Zhu, Li-Ping; Xu, You-Yi

    2015-06-15

    Here we describe the development of versatile antifouling polyethersulfone (PES) filtration membranes modified via surface grafting of zwitterionic polymers from a reactive amphiphilic copolymer additive. Amphiphilic polyethersulfone-block-poly(2-hydroxyethyl methacrylate) (PES-b-PHEMA) was beforehand designed and used as the blending additive of PES membranes prepared by phase inversion technique. The surface enriched PHEMA blocks on membrane surface acted as an anchor to immobilize the initiating site. Poly(sulfobetaine methacrylate) (PSBMA) were subsequently grafted onto the PES blend membranes by surface-initiated atom transfer radical polymerization (SI-ATRP). The analysis of surface chemistry confirmed the successful grafting of zwitterionic PSBMA brushes on PES membrane surface. The resulted PES-g-PSBMA membranes were capable of separating proteins from protein solution and oil from oil/water emulsion efficiently. Furthermore, the modified membranes showed high hydrophilicity and strongly antifouling properties due to the incorporation of well-defined PSBMA layer. In addition, the PES-g-PSBMA membranes exhibited excellent blood compatibility and durability during the washing process. The developed antifouling PES membranes are versatile and can find their applications in protein filtration, blood purification and oil/water separation, etc. Copyright © 2015 Elsevier Inc. All rights reserved.

  19. Toxicity following laundry detergent pod ingestion.

    PubMed

    Schneir, Aaron B; Rentmeester, Landen; Clark, Richard F; Cantrell, F Lee

    2013-06-01

    Laundry detergent pods (LDPs) have only recently become available in the United States, and there has been increasing concern regarding pediatric ingestions of them. We describe a 15-month-old female infant who ingested an LDP and had a depressed level of consciousness, metabolic acidosis, pulmonary toxicity, and swallowing difficulties. It is currently unclear what the exact etiologic agent(s) is responsible for the toxicity associated with LDPs. The case demonstrates the potential for significant toxicity following the ingestion of an LDP. Clearly, measures should be taken to avoid ingestions of these products.

  20. Effects of Lipid-Analog Detergent Solubilization on the Functionality and Lipidic Cubic Phase Mobility of the Torpedo californica Nicotinic Acetylcholine Receptor

    PubMed Central

    Padilla-Morales, Luis F.; Morales-Pérez, Claudio L.; De La Cruz-Rivera, Pamela C.; Asmar-Rovira, Guillermo; Báez-Pagán, Carlos A.

    2011-01-01

    Over the past three decades, the Torpedo californica nicotinic acetylcholine receptor (nAChR) has been one of the most extensively studied membrane protein systems. However, the effects of detergent solubilization on nAChR stability and function are poorly understood. The use of lipid-analog detergents for nAChR solubilization has been shown to preserve receptor stability and functionality. The present study used lipid-analog detergents from phospholipid-analog and cholesterol-analog detergent families for solubilization and affinity purification of the receptor and probed nAChR ion channel function using planar lipid bilayers (PLBs) and stability using analytical size exclusion chromatography (A-SEC) in the detergent-solubilized state. We also examined receptor mobility on the lipidic cubic phase (LCP) by measuring the nAChR mobile fraction and diffusion coefficient through fluorescence recovery after photobleaching (FRAP) experiments using lipid-analog and non-lipid-analog detergents. Our results show that it is possible to isolate stable and functional nAChRs using lipid-analog detergents, with characteristic ion channel currents in PLBs and minimal aggregation as observed in A-SEC. Furthermore, fractional mobility and diffusion coefficient values observed in FRAP experiments were similar to the values observed for these parameters in the recently LCP-crystallized β2-adrenergic receptor. The overall results show that phospholipid-analog detergents with 16 carbon acyl-chains support nAChR stability, functionality and LCP mobility. PMID:21922299

  1. Determining the Residual Characteristics of Alkylphenols, Arsenic, and Lead as well as Assessing the Exposures of 1,4-Dioxane from Household Food Detergents.

    PubMed

    Lin, Wan-Ting; Chen, Wun-Ling; Cheng, Wei-Chih; Chang, Hui-Chuan; Tsai, Shih-Wei

    2017-07-01

    Food detergents are commonly used in households. The main components of detergents include surfactants and water. It has been found that certain compounds, which may cause health concerns, appear in food detergents, such as alkylphenols, arsenic, and lead. After applying food detergents is when people may be exposed to various levels of these chemicals when there are residues. In this study, the Taguchi experimental design was performed to determine the possible factors that might affect the residual characteristics of food detergent on dishware, fruits, and vegetables. The results showed that the variety of detergent was found to be the most significant factor affecting the residue amounts of arsenic (62.9%) and lead (71.6%) on fruits and vegetables, whereas the concentration of detergent used affected the amount of lead residue only (10.5%). On the other hand, dishware material, the concentration of analytes, immersion time, and type of surfactant contributed to arsenic residues on dishware, whereas technical nonylphenol isomer residues on dishware increased as the concentration of spiked analyte increased. In addition, the occurrence of 1,4-dioxane, a possible human carcinogen, in household food detergents in Taiwan was also determined in this research by solid-phase microextraction and GC-MS. Among the 80 detergent samples, 71 contained different concentrations ranges of 1,4-dioxane, from 0.03 to ~3.73 µg/g. In the exposure assessment, it was estimated that the maximum amounts of 1,4-dioxane in contact with the skin from the use of household food detergent in Taiwan was 0.015 µg/kg/day.

  2. Development of the volunteer peer educator role in a community Cardiovascular Health Awareness Program (CHAP): a process evaluation in two communities.

    PubMed

    Karwalajtys, Tina; McDonough, Beatrice; Hall, Heather; Guirguis-Younger, Manal; Chambers, Larry W; Kaczorowski, Janusz; Lohfeld, Lynne; Hutchison, Brian

    2009-08-01

    Volunteers can support the delivery and sustainability of programs promoting chronic disease awareness to improve health at the community level. This paper describes the development of the peer education component of the Cardiovascular Health Awareness Program (CHAP) and assessment of the volunteer peer educator role in a community-wide demonstration project in two mid-sized Ontario communities. A case study approach was used incorporating process learning, a volunteer survey and debriefing discussions with volunteers. A post-program questionnaire was administered to 48 volunteers. Five debriefing discussions were conducted with 27 volunteers using a semi-structured interview guide. Discussions were audio-recorded and transcribed. Analysis used an editing approach to identify themes, taking into account the community-specific context. Volunteers reported an overall positive experience and identified rewarding aspects of their involvement. They felt well prepared but appreciated ongoing training and support and requested more refresher training. Understanding of program objectives increased volunteer satisfaction. Volunteers continued to develop their role during the program; however, organizational and logistical factors sometimes limited skill acquisition and contributions. The prospect of greater involvement in providing tailored health education resources addressing modifiable risk factors was acceptable to most volunteers. Continued refinement of strategies to recruit, train, retain and support volunteers strengthened the peer education component of CHAP. The experience and contributions of volunteers were influenced by the wider context of program delivery. Process evaluation allowed program planners to anticipate challenges, strengthen support for volunteer activities, and expand the peer educator role. This learning can inform similar peer-led health promotion initiatives.

  3. Arsenic in detergents: Possible danger and pollution hazard

    USGS Publications Warehouse

    Angino, E.E.; Magnuson, L.M.; Waugh, T.C.; Galle, O.K.; Bredfeldt, J.

    1970-01-01

    Arsenic at a concentration of 10 to 70 parts per million has been detected in several common presoaks and household detergents. Arsenic values of 2 to 8 parts per billion have been measured in the Kansas River. These concentrations are close to the amount (10 parts per billion) recommended by the United States Public Health Service as a drinking-water standard.

  4. Preparation and evaluation of a silica-based 1-alkyl-3-(propyl-3-sulfonate) imidazolium zwitterionic stationary phase for high-performance liquid chromatography.

    PubMed

    Qiu, Hongdeng; Jiang, Qiong; Wei, Zheng; Wang, Xusheng; Liu, Xia; Jiang, Shengxiang

    2007-09-07

    A new zwitterionic stationary phase based on silica bonded with 1-alkyl-3-(propyl-3-sulfonate) imidazolium was synthesized and characterized in this paper. The materials have been confirmed and evaluated by elemental analysis, thermogravimetric analysis and X-ray photoelectron spectroscopy. Potassium and calcium were separated simultaneously with several common inorganic anions including an iodate, chloride, bromide, nitrate and iodide on the phase. The effects of the concentration, organic solvent and pH of the eluent on the separation of anions were studied. Operated in the anion-exchange mode, this new stationary phase shows considerable promise for the separation of anions. Bases, vitamins and three imidazolium ionic liquids with different alkyl chains are also separated successfully on this column. The stationary phase has multiple retention mechanisms, such as anion-exchange, electrostatic attraction and repulsion interactions, and hydrophobic interaction between the zwitterionic stationary phase and specimens.

  5. Laundry detergent "pod" ingestions: a case series and discussion of recent literature.

    PubMed

    Beuhler, Michael C; Gala, Payal K; Wolfe, Heather A; Meaney, Peter A; Henretig, Fred M

    2013-06-01

    The objectives of this study were to present and explore the clinical presentation of the increasingly common pediatric exposure to the widely available single-use laundry packets or "laundry pods." This is a case report of 4 pediatric patients with significant toxicity due to laundry pod detergent exposure and a review of the available literature including abstract-only publications. An unexpectedly severe clinical pattern was noted; 3 of the 4 children required intubation for management, airway injury was noted in 1 of them, and 2 of them had hospital courses of at least 1 week. The literature suggests that laundry pod exposures are associated with increased morbidity compared to traditional laundry detergent exposures. To date, no specific contaminant or component has been identified as being responsible for the injury, although some evidence points to the surfactant component. A different approach to the triage and management of pediatric exposures to laundry detergent pod ingestions is required compared with nonpod ingestions. Although the exact cause is not known, practitioners should be vigilant for rapid onset of neurological impairment and inability to protect the airway in addition to its caustic effects.

  6. Metabolism of Some Anionic Tallow-based Detergents by Sewage Microorganisms1

    PubMed Central

    Cordon, Theone C.; Maurer, Elmer W.; Nuñez-Ponzoa, M. V.; Stirton, A. J.

    1968-01-01

    A method in which the test detergent was the sole source of carbon was used to study the metabolism of several tallow-based detergents. These were tallow alcohol sulfates, long-chain ether alcohol sulfates, and esters of α-sulfo fatty acids. Sodium p-(1-methylundecyl)benzenesulfonate (LAS) was used as a reference material. The alcohol sulfates were the most rapidly and completely metabolized (96 to 99%), and one ether alcohol sulfate was 94% degraded. The other compounds were metabolized to the extent of 61 to 87%; LAS was 80% degraded. Except for the alcohol sulfates, loss of methylene blue activity (MBAS) occurred long before the chemical oxygen demand (COD) values had reached a minimum; with the alcohol sulfates, MBAS and COD decreased simultaneously. PMID:5636472

  7. GDP-GTP exchange processes of G{alpha}i1 protein are accelerated/decelerated depending on the type and the concentration of added detergents.

    PubMed

    Kubota, Makoto; Tanaka, Takeshi; Kohno, Toshiyuki; Wakamatsu, Kaori

    2009-12-01

    Although detergents have been widely used in G-protein studies to increase solubility and stability of the protein, we noticed that detergents modulate the nucleotide-binding properties of G-proteins. Hence, we analysed the effects of detergents on guanine nucleotide exchange reactions of Galpha(i1). Lubrol PX, a non-ionic detergent, which has been widely used in nucleotide dissociation/binding assays, was found to accelerate both GDP dissociation and GTPgammaS binding from/to Galpha in parallel at above its critical micelle concentration (cmc). Sodium cholate, an anionic detergent, which have been used to extract G-proteins from animal tissues, decelerated and accelerated GDP dissociation below and above its cmc, respectively. Surprisingly, micellar cholate decelerated GTPgammaS binding, and the binding rate constant was decreased by three orders of magnitude in the presence of 2% cholate. These results demonstrate that the guanine nucleotide exchange reactions of Galpha(i1) are drastically modulated by detergents differently depending on the type and the state (monomeric or micellar) of the detergents and that dissociation of GDP from Galpha(i1) does not necessarily lead to immediate binding of GTP to Galpha(i1) in some cases. These effects of detergents on G-proteins must be taken into account in G-protein experiments.

  8. Fatty acid profiles from the plasma membrane and detergent resistant membranes of two plant species.

    PubMed

    Carmona-Salazar, Laura; El Hafidi, Mohammed; Gutiérrez-Nájera, Nora; Noyola-Martínez, Liliana; González-Solís, Ariadna; Gavilanes-Ruíz, Marina

    2015-01-01

    It is essential to establish the composition of the plant plasma membrane in order to understand its organization and behavior under continually changing environments. Knowledge of the lipid phase, in particular the fatty acid (FA) complex repertoire, is important since FAs determine many of the physical-chemical membrane properties. FAs are constituents of the membrane glycerolipid and sphingolipid backbones and can also be linked to some sterols. In addition, FAs are components of complex lipids that can constitute membrane micro-domains, and the use of detergent-resistant membranes is a common approach to study their composition. The diversity and cellular allocation of the membrane lipids containing FAs are very diverse and the approaches to analyze them provide only general information. In this work, a detailed FA analysis was performed using highly purified plasma membranes from bean leaves and germinating maize embryos and their respective detergent-resistant membrane preparations. The analyses showed the presence of a significant amount of very long chain FAs (containing 28C, 30C and 32C), in both plasma membrane preparations from bean and maize, that have not been previously reported. Herein is demonstrated that a significant enrichment of very long chain saturated FAs and saturated FAs can occur in detergent-resistant membrane preparations, as compared to the plasma membranes from both plant species. Considering that a thorough analysis of FAs is rarely performed in purified plasma membranes and detergent-resistant membranes, this work provides qualitative and quantitative evidence on the contributions of the length and saturation of FAs to the organization of the plant plasma membrane and detergent-resistant membranes. Copyright © 2014 Elsevier Ltd. All rights reserved.

  9. Inclusion of detergent in a cleaning regime and effect on microbial load in livestock housing

    PubMed Central

    Hancox, L. R.; Le Bon, M.; Dodd, C. E. R.; Mellits, K. H.

    2013-01-01

    Determining effective cleaning and disinfection regimes of livestock housing is vital to improving the health of resident animals and reducing zoonotic disease. A cleaning regime consisting of scraping, soaking with or without detergent (treatment and control), pressure washing, disinfection and natural drying was applied to multiple pig pens. After each cleaning stage, samples were taken from different materials and enumerated for total aerobic count (TAC) and Enterobacteriaceae (ENT). Soaking with detergent (Blast-Off, Biolink) caused significantly greater reductions of TAC and ENT on metal, and TAC on concrete, compared with control. Disinfection effect (Virkon S, DuPont) was not significantly associated with prior detergent treatment. Disinfection significantly reduced TAC and ENT on concrete and stock board but not on metal. Twenty-four hours after disinfection TAC and ENT on metal and stock board were significantly reduced, but no significant reductions occurred in the subsequent 96 hours. Counts on concrete did not significantly reduce during the entire drying period (120 hours). Detergent and disinfectant have varying bactericidal effects according to the surface and bacterial target; however, both can significantly reduce microbial numbers so should be used during cleaning, with a minimum drying period of 24 hours, to lower bacterial counts effectively. PMID:23839725

  10. Inclusion of detergent in a cleaning regime and effect on microbial load in livestock housing.

    PubMed

    Hancox, L R; Le Bon, M; Dodd, C E R; Mellits, K H

    Determining effective cleaning and disinfection regimes of livestock housing is vital to improving the health of resident animals and reducing zoonotic disease. A cleaning regime consisting of scraping, soaking with or without detergent (treatment and control), pressure washing, disinfection and natural drying was applied to multiple pig pens. After each cleaning stage, samples were taken from different materials and enumerated for total aerobic count (TAC) and Enterobacteriaceae (ENT). Soaking with detergent (Blast-Off, Biolink) caused significantly greater reductions of TAC and ENT on metal, and TAC on concrete, compared with control. Disinfection effect (Virkon S, DuPont) was not significantly associated with prior detergent treatment. Disinfection significantly reduced TAC and ENT on concrete and stock board but not on metal. Twenty-four hours after disinfection TAC and ENT on metal and stock board were significantly reduced, but no significant reductions occurred in the subsequent 96 hours. Counts on concrete did not significantly reduce during the entire drying period (120 hours). Detergent and disinfectant have varying bactericidal effects according to the surface and bacterial target; however, both can significantly reduce microbial numbers so should be used during cleaning, with a minimum drying period of 24 hours, to lower bacterial counts effectively.

  11. Preclinical manufacture of anti-HER2 liposome-inserting, scFv-PEG-lipid conjugate. 2. Conjugate micelle identity, purity, stability, and potency analysis.

    PubMed

    Nellis, David F; Giardina, Steven L; Janini, George M; Shenoy, Shilpa R; Marks, James D; Tsai, Richard; Drummond, Daryl C; Hong, Keelung; Park, John W; Ouellette, Thomas F; Perkins, Shelley C; Kirpotin, Dmitri B

    2005-01-01

    Analytical methods optimized for micellar F5cys-MP-PEG(2000)-DPSE protein-lipopolymer conjugate are presented. The apparent micelle molecular weight, determined by size exclusion chromatography, ranged from 330 to 960 kDa. The F5cys antibody and conjugate melting points, determined by differential scanning calorimetry, were near 82 degrees C. Traditional methods for characterizing monodisperse protein species were inapplicable to conjugate analysis. The isoelectric point of F5cys (9.2) and the conjugate (8.9) were determined by capillary isoelectric focusing (cIEF) after addition of the zwitterionic detergent CHAPS to the buffer. Conjugate incubation with phospholipase B selectively removed DSPE lipid groups and dispersed the conjugate prior to separation by chromatographic methods. Alternatively, adding 2-propanol (29.4 vol %) and n-butanol (4.5 vol %) to buffers for salt-gradient cation exchange chromatography provided gentler, nonenzymatic dispersion, resulting in well-resolved peaks. This method was used to assess stability, identify contaminants, establish lot-to-lot comparability, and determine the average chromatographic purity (93%) for conjugate lots, described previously. The F5cys amino acid content was confirmed after conjugation. The expected conjugate avidity for immobilized HER-2/neu was measured by bimolecular interaction analysis (BIAcore). Mock therapeutic assemblies were made by conjugate insertion into preformed doxorubicin-encapsulating liposomes for antibody-directed uptake of doxorubicin by HER2-overexpressing cancer cells in vitro. Together these developed assays established that the manufacturing method as described in the first part of this study consistently produced F5cys-MP-PEG(2000)-DSPE having sufficient purity, stability, and functionality for use in preclinical toxicology investigations.

  12. Affinity purification of angiotensin type 2 receptors from N1E-115 cells: evidence for agonist-induced formation of multimeric complexes.

    PubMed

    Siemens, I R; Yee, D K; Reagan, L P; Fluharty, S J

    1994-01-01

    The murine neuroblastoma N1E-115 cell line possesses type 1 and type 2 angiotensin II (AngII) receptor subtypes. In vitro differentiation of these cells substantially increases the density of the AT2-receptor subtype, whereas the density of the AT1 receptors remains unchanged. In the present study, we report that the zwitterionic detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) selectively solubilized AT2 receptors from N1E-115 cell membranes and that these receptors could be purified further to near homogeneity by affinity chromatography. More specifically, the presence of an agonist (AngII) during affinity purification of AT2 receptors resulted in the elution of high (110-kDa) and low (66-kDa) molecular mass proteins as determined by gel electrophoresis under nonreducing conditions. In contrast, when the nonselective antagonist Sar1,Ile8-AngII was used during purification, only the lower 66-kDa protein was observed. Affinity purification in the presence of the peptide and nonpeptide AT2-receptor antagonists CGP42112A and PD123319 also resulted in elution of the same 66-kDa protein, but unlike that in the presence of Sar1,Ile8-AngII, some of the high molecular weight site was observed as well. On the other hand, Losartan, an AT1-receptor antagonist, was completely ineffective in eluting any AngII receptors from the affinity column, further confirming their AT2 identity. After agonist elution, the 110-kDa band dissociated into two low molecular mass bands of 66 kDa and 54 kDa when sodium dodecyl sulfate-gel electrophoresis was run under reducing conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

  13. Biodegradation of a keratin waste and the concomitant production of detergent stable serine proteases from Paecilomyces lilacinus.

    PubMed

    Cavello, I A; Cavalitto, S F; Hours, R A

    2012-07-01

    Paecilomyces lilacinus (LPS 876) efficiently degraded keratin in chicken feather during submerged cultivation producing extracellular proteases. Characterization of crude protease activity was done including its compatibility in commercial detergents. Optimum pH and temperature were 10.0 and 60 °C, respectively. Protease activity was enhanced by Ca²⁺ but was strongly inhibited by PMSF and by Hg²⁺ suggesting the presence of thiol-dependent serine proteases. The crude protease showed extreme stability toward non-ionic (Tween 20, Tween 85, and Triton X-100) and anionic (SDS) surfactants, and relative stability toward oxidizing agent (H₂O₂ and sodium perborate). In addition, it showed excellent stability and compatibility with various solid and liquid commercial detergents from 30 to 50 °C. The enzyme preparation retained more than 95% of its initial activity with solid detergents (Ariel™ and Drive™) and 97% of its original activity with a liquid detergent (Ace™) after pre-incubation at 40 °C. The protective effect of polyols (propylene glycol, PEG 4000, and glycerol) on the heat inactivation was also examined and the best results were obtained with glycerol from 50 to 60 °C. Considering its promising properties, P. lilacinus enzymatic preparation may be considered as a candidate for use in biotechnological processes (i.e., as detergent additive) and in the processing of keratinous wastes.

  14. Biochemical process of low level radioactive liquid simulation waste containing detergent

    NASA Astrophysics Data System (ADS)

    Kundari, Noor Anis; Putra, Sugili; Mukaromah, Umi

    2015-12-01

    Research of biochemical process of low level radioactive liquid waste containing detergent has been done. Thse organic liquid wastes are generated in nuclear facilities such as from laundry. The wastes that are cotegorized as hazard and poison materials are also radioactive. It must be treated properly by detoxification of the hazard and decontamination of the radionuclides to ensure that the disposal of the waste meets the requirement of standard quality of water. This research was intended to determine decontamination factor and separation efficiensies, its kinetics law, and to produce a supernatant that ensured the environmental quality standard. The radioactive element in the waste was thorium with activity of 5.10-5 Ci/m3. The radioactive liquid waste which were generated in simulation plant contains detergents that was further processed by aerobic biochemical process using SGB 103 bacteria in a batch reactor equipped with aerators. Two different concentration of samples were processed and analyzed for 212 hours and 183 hours respectively at a room temperature. The product of this process is a liquid phase called as supernatant and solid phase material called sludge. The chemical oxygen demand (COD), biological oxygen demand (BOD), suspended solid (SS), and its alpha activity were analyzed. The results show that the decontamination factor and the separation efficiency of the lower concentration samples are higher compared to the samples with high concentration. Regarding the decontamination factor, the result for 212 hours processing of waste with detergent concentration of 1.496 g/L was 3.496 times, whereas at the detergent concentration of 0.748 g/L was 15.305 times for 183 hours processing. In case of the separation efficiency, the results for both samples were 71.396% and 93.465% respectively. The Bacterial growth kinetics equation follow Monod's model and the decreasing of COD and BOD were first order with the rate constant of 0.01 hour-1.

  15. Biochemical process of low level radioactive liquid simulation waste containing detergent

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

    Kundari, Noor Anis, E-mail: nooranis@batan.go.id; Putra, Sugili; Mukaromah, Umi

    Research of biochemical process of low level radioactive liquid waste containing detergent has been done. Thse organic liquid wastes are generated in nuclear facilities such as from laundry. The wastes that are cotegorized as hazard and poison materials are also radioactive. It must be treated properly by detoxification of the hazard and decontamination of the radionuclides to ensure that the disposal of the waste meets the requirement of standard quality of water. This research was intended to determine decontamination factor and separation efficiensies, its kinetics law, and to produce a supernatant that ensured the environmental quality standard. The radioactive elementmore » in the waste was thorium with activity of 5.10{sup −5} Ci/m{sup 3}. The radioactive liquid waste which were generated in simulation plant contains detergents that was further processed by aerobic biochemical process using SGB 103 bacteria in a batch reactor equipped with aerators. Two different concentration of samples were processed and analyzed for 212 hours and 183 hours respectively at a room temperature. The product of this process is a liquid phase called as supernatant and solid phase material called sludge. The chemical oxygen demand (COD), biological oxygen demand (BOD), suspended solid (SS), and its alpha activity were analyzed. The results show that the decontamination factor and the separation efficiency of the lower concentration samples are higher compared to the samples with high concentration. Regarding the decontamination factor, the result for 212 hours processing of waste with detergent concentration of 1.496 g/L was 3.496 times, whereas at the detergent concentration of 0.748 g/L was 15.305 times for 183 hours processing. In case of the separation efficiency, the results for both samples were 71.396% and 93.465% respectively. The Bacterial growth kinetics equation follow Monod’s model and the decreasing of COD and BOD were first order with the rate constant of

  16. Structural analysis of zwitterionic liquids vs. homologous ionic liquids

    NASA Astrophysics Data System (ADS)

    Wu, Boning; Kuroda, Kosuke; Takahashi, Kenji; Castner, Edward W.

    2018-05-01

    Zwitterionic liquids (Zw-ILs) have been developed that are homologous to monovalent ionic liquids (ILs) and show great promise for controlled dissolution of cellulosic biomass. Using both high energy X-ray scattering and atomistic molecular simulations, this article compares the bulk liquid structural properties for novel Zw-ILs with their homologous ILs. It is shown that the significant localization of the charges on Zw-ILs leads to charge ordering similar to that observed for conventional ionic liquids with monovalent anions and cations. A low-intensity first sharp diffraction peak in the liquid structure factor S(q) is observed for both the Zw-IL and the IL. This is unexpected since both the Zw-IL and IL have a 2-(2-methoxyethoxy)ethyl (diether) functional group on the cationic imidazolium ring and ether functional groups are known to suppress this peak. Detailed analyses show that this intermediate range order in the liquid structure arises for slightly different reasons in the Zw-IL vs. the IL. For the Zw-IL, the ether tails in the liquid are shown to aggregate into nanoscale domains.

  17. Laundry detergent pod ingestions: is there a need for endoscopy?

    PubMed

    Smith, Erika; Liebelt, Erica; Nogueira, Jan

    2014-09-01

    Laundry detergent pod (LDP) exposures in children have resulted in several referrals to the emergency department. Signs and symptoms can include gastrointestinal symptoms (vomiting, drooling), neurological symptoms (depressed sensorium), or metabolic changes (lactic acidosis). There is limited literature on esophageal injury following LDP ingestions. We reviewed three cases of pediatric LDP ingestions that underwent an upper endoscopy in a tertiary care pediatric hospital. All of our patients were younger than 3 years old. The upper endoscopies revealed superficial esophageal erosions in two patients and erythema in the other. None of the patients had oral burns. Two of them developed swallowing dysfunction. Follow-up upper GI studies were normal. Our three patients ingested laundry detergent pods and all of them developed some degree of esophageal injury despite the absence of oral erythema, ulcers, or swelling. A review of literature suggests LDP exposures are more severe than non-pod detergents. Reasons as to why this may be remain unclear, although investigation into the ingredients and mode of delivery may help us to better understand. In a literature review, no esophageal strictures have been reported after LDP ingestion. We reviewed esophageal injury classification systems in an attempt to predict who may be at greatest risk for stricture based on initial findings. Our case series demonstrates it is hard to predict esophageal injury based on signs and symptoms. Based on a literature review, long-term esophageal stricture is unlikely, but if gastrointestinal symptoms persist, it is reasonable to evaluate with an upper endoscopy. Larger studies are needed.

  18. Active Detergent-solubilized H+,K+-ATPase Is a Monomer*

    PubMed Central

    Dach, Ingrid; Olesen, Claus; Signor, Luca; Nissen, Poul; le Maire, Marc; Møller, Jesper V.; Ebel, Christine

    2012-01-01

    The H+,K+-ATPase pumps protons or hydronium ions and is responsible for the acidification of the gastric fluid. It is made up of an α-catalytic and a β-glycosylated subunit. The relation between cation translocation and the organization of the protein in the membrane are not well understood. We describe here how pure and functionally active pig gastric H+,K+-ATPase with an apparent Stokes radius of 6.3 nm can be obtained after solubilization with the non-ionic detergent C12E8, followed by exchange of C12E8 with Tween 20 on a Superose 6 column. Mass spectroscopy indicates that the β-subunit bears an excess mass of 9 kDa attributable to glycosylation. From chemical analysis, there are 0.25 g of phospholipids and around 0.024 g of cholesterol bound per g of protein. Analytical ultracentrifugation shows one main complex, sedimenting at s20,w = 7.2 ± 0.1 S, together with minor amounts of irreversibly aggregated material. From these data, a buoyant molecular mass is calculated, corresponding to an H+,K+-ATPase α,β-protomer of 147.3 kDa. Complementary sedimentation velocity with deuterated water gives a picture of an α,β-protomer with 0.9–1.4 g/g of bound detergent and lipids and a reasonable frictional ratio of 1.5, corresponding to a Stokes radius of 7.1 nm. An α2,β2 dimer is rejected by the data. Light scattering coupled to gel filtration confirms the monomeric state of solubilized H+,K+-ATPase. Thus, α,β H+,K+-ATPase is active at least in detergent and may plausibly function as a monomer, as has been established for other P-type ATPases, Ca2+-ATPase and Na+,K+-ATPase. PMID:23055529

  19. Respiratory symptoms and peak expiratory flow rates in workers of a Nigerian soap and detergent industry.

    PubMed

    Bamidele, J O

    2002-01-01

    This comparative cross-sectional study was carried out to assess the respiratory symptoms and peak expiratory flow rates of the factory(study group) and office(control group) workers in a soap and detergent industry in Ilorin in relation to the occupational hazards of chemical fumes and detergent dust in the industry. Upper respiratory tract infections were found in 67.5% and 10.6% of the study group and control group respectively. The study shows that the factory workers experienced hazards (e.g. chemical fumes and detergent dust) at work more than the office workers. Personal protective devices such as boots, face masks, gloves, earmuffs and goggles were not consistently used since they were inadequate in supply, worn out and of substandard qualities. The general reduction in the mean values of peak expiratory flow rate in the factory workers than in the office workers as observed in this study may possibly, be a pointer to the effect of industrial exposure to chemical fumes and detergent dust over the years. There is the need to follow up these workers in order to detect early any possible disease and complications that may arise.

  20. Proteomic determination of widespread detergent-insolubility including Abeta but not tau early in the pathogenesis of Alzheimer's disease.

    PubMed

    Woltjer, Randall L; Cimino, P J; Boutté, Angela M; Schantz, Aimee M; Montine, Kathleen S; Larson, Eric B; Bird, Thomas; Quinn, Joseph F; Zhang, Jing; Montine, Thomas J

    2005-11-01

    Biochemical characterization of the major detergent-insoluble proteins that comprise hallmark histopathologic lesions initiated the molecular era of Alzheimer's disease (AD) research. Here, we reinvestigated detergent-insoluble proteins in AD using modern proteomic techniques. Using liquid chromatography (LC)-mass spectrometry (MS)-MS-based proteomics, we robustly identified 125 proteins in the detergent-insoluble fraction of late-onset AD (LOAD) temporal cortex that included several proteins critical to Abeta production, components of synaptic scaffolding, and products of genes linked to an increased risk of LOAD; we verified 15 of 15 of these proteins by Western blot. Following multiple analyses, we estimated that these represent ~80% of detergent-insoluble proteins in LOAD detectable by our method. Abeta, tau, and 7 of 8 other newly identified detergent-insoluble proteins were disproportionately increased in temporal cortex from patients with LOAD and AD derived from mutations in PSEN1 and PSEN2; all of these except tau were elevated in individuals with prodromal dementia, while none except Abeta were elevated in aged APPswe mice. These results are consistent with the amyloid hypothesis of AD and extend it to include widespread protein insolubility, not exclusively Abeta insolubility, early in AD pathogenesis even before the onset of clinical dementia.

  1. Cholesterol depletion modulates detergent resistant fraction of human serotonin(1A) receptors.

    PubMed

    Sahu, Santosh Kumar; Saxena, Roopali; Chattopadhyay, Amitabha

    2012-11-01

    Insolubility of membrane components in non-ionic detergents such as Triton X-100 at low temperature is a widely used biochemical criterion to identify, isolate and characterize membrane domains. In this work, we monitored the detergent insolubility of the serotonin(1A) receptor in CHO cell membranes and its modulation by membrane cholesterol. The serotonin(1A) receptor is an important member of the G-protein coupled receptor family. It is implicated in the generation and modulation of various cognitive, behavioral and developmental functions and serves as a drug target. Our results show that a significant fraction (∼28%) of the serotonin(1A) receptor resides in detergent-resistant membranes (DRMs). Interestingly, the fraction of the serotonin(1A) receptor in DRMs exhibits a reduction upon membrane cholesterol depletion. In addition, we show that contents of DRM markers such as flotillin-1, caveolin-1 and GM₁ are altered in DRMs upon cholesterol depletion. These results assume significance since the function of the serotonin(1A) receptor has previously been shown to be affected by membrane lipids, specifically cholesterol. Our results are relevant in the context of membrane organization of the serotonin(1A) receptor in particular, and G-protein coupled receptors in general.

  2. A comparison of decontamination effects of commercially available detergents in rats pre-exposed to topical sulphur mustard.

    PubMed

    Misik, Jan; Jost, Petr; Pavlikova, Ruzena; Vodakova, Eva; Cabal, Jiri; Kuca, Kamil

    2013-06-01

    The genotoxic vesicant sulphur mustard [bis-2-(chloroethyl)sulphide] is a chemical warfare agent which is easily available due to its relatively simple synthesis. Thus, sulphur mustard is a potential agent for mass contamination. In this study, we focused on sulphur mustard toxicity and decontamination in a rat model using commercially available detergent mixtures for dermal decontamination. Male Wistar rats were percutaneously treated with sulphur mustard and subjected to wet decontamination 2 min postexposure. Commercially produced detergents Neodekont™, Argos™, Dermogel™ and FloraFree™ were tested for their decontamination efficacy against an exposed group and their protective ratios determined. The results showed that all tested detergent solutions produced an increase in the median lethal dose [LD(50) = 9.83 (5.87-13.63) mg·kg(-1)] in comparison to controls, which led to increased survival of experimental animals. In general, all tested detergents provided modest decontamination efficacy (PR = 2.0-5.7). The highest protective ratio (5.7) was consistently achieved with Argos™. Accordingly, Argos™ should be considered in further investigation of mass casualty decontamination.

  3. Stabilization of the predominant disease-causing aldolase variant (A149P) with zwitterionic osmolytes.

    PubMed

    Stopa, Jack D; Chandani, Sushil; Tolan, Dean R

    2011-02-08

    Hereditary fructose intolerance (HFI) is a disease of carbohydrate metabolism that can result in hyperuricemia, hypoglycemia, liver and kidney failure, coma, and death. Currently, the only treatment for HFI is a strict fructose-free diet. HFI arises from aldolase B deficiency, and the most predominant HFI mutation is an alanine to proline substitution at position 149 (A149P). The resulting aldolase B with the A149P substitution (AP-aldolase) has activity that is <100-fold that of the wild type. The X-ray crystal structure of AP-aldolase at both 4 and 18 °C reveals disordered adjacent loops of the (α/β)(8) fold centered around the substitution, which leads to a dimeric structure as opposed to the wild-type tetramer. The effects of osmolytes were tested for restoration of structure and function. An initial screen of osmolytes (glycerol, sucrose, polyethylene glycol, 2,4-methylpentanediol, glutamic acid, arginine, glycine, proline, betaine, sarcosine, and trimethylamine N-oxide) reveals that glycine, along with similarly structured compounds, betaine and sarcosine, protects AP-aldolase structure and activity from thermal inactivation. The concentration and functional moieties required for thermal protection show a zwitterion requirement. The effects of osmolytes in restoring structure and function of AP-aldolase are described. Testing of zwitterionic osmolytes of increasing size and decreasing fractional polar surface area suggests that osmolyte-mediated AP-aldolase stabilization occurs neither primarily through excluded volume effects nor through transfer free energy effects. These data suggest that AP-aldolase is stabilized by binding to the native structure, and they provide a foundation for developing stabilizing compounds for potential therapeutics for HFI.

  4. Preliminary characterization of Thy-1.1 and Ag-B antigens from rat tissues solubilized in detergents

    PubMed Central

    Letarte-Muirhead, Michelle; Acton, Ronald T.; Williams, Alan F.

    1974-01-01

    1. A radioactive binding assay for Thy-1.1 alloantigen which functions in the presence of detergents was established by using glutaraldehyde-fixed thymocytes as target cells. Thy-1.1 activity in detergent extracts was then assayed by measuring inhibition of the binding assay. 2. Solubilization of Thy-1.1 from whole thymocytes, and their membranes by a large number of non-ionic detergents and deoxycholate was studied. In the same extracts Ag-B(4) histocompatibility antigenic activities were measured. With the exception of Nonidet P-40, the detergents did not affect the antigenicity of Thy-1.1, but only Lubrol-PX and deoxycholate gave effective solubilization as measured by activity remaining in the supernatant after centrifugation at 200000g for 40min. With Ag-B(4) antigen, Triton X-100, Triton X-67 and Nonidet P-40 gave effective solubilization as well as Lubrol-PX and deoxycholate. Solubilization of Thy-1.1 activity from leukaemia cells and a brain homogenate was also studied, but none of the non-ionic detergents gave satisfactory results with these tissues. 3. Extracts from thymocyte membranes were further examined by gel filtration and sucrose gradient centrifugation. The Thy-1.1 activity behaved as a single component in deoxycholate with a density similar to that of a globular protein, but in Lubrol-PX the antigen was contained in a low-density complex. In Lubrol-PX extracts Ag-B(4) was also found in aggregates not observed in deoxycholate. 4. The s20,w values for Thy-1.1 and Ag-B(4) antigens in deoxycholate were 2.4 and 4.4, and v̄ values were 0.70 and 0.75 respectively. The Stokes radius observed for Thy-1.1 was 3.1nm and for Ag-B(4) 5.3nm. By using these values the molecular weights for the antigen–detergent complexes were calculated to be 28000 for Thy-1.1 and 100000 for Ag-B(4). PMID:4219284

  5. Effect of detergents on the H(+)-ATPase activity of inside-out and right-side-out plant plasma membrane vesicles.

    PubMed

    Palmgren, M G; Sommarin, M; Ulvskov, P; Larsson, C

    1990-01-29

    In search for a detergent to be used to assess the sidedness of plant plasma membrane vesicles by enzyme latency we tested the effect of 42 detergents on the ATPase activity of right-side-out and inside-out plasma membrane vesicles from sugar beet leaves. Most of the detergents seemed to inactivate the ATPase in addition to disrupting the permeability barrier to ATP. There were two main exceptions, namely long chain polyoxyethylene acyl ethers, such as detergents of the Brij series and Lubrol WX, and long chain lysophospholipids. These two types of detergents permeabilized the membranes at low concentrations and did not inhibit the ATPase at higher concentrations. Unmasking of latent active sites seemed to explain the activation of the plasma membrane H(+)-ATPase produced by long chain polyoxyethylene acyl ethers. These detergents should therefore be ideal for determination of vesicle orientation based on ATPase latency. By contrast, long chain lysophospholipids were found to be highly specific activators of the enzyme. In addition, long chain fatty acids were found to strongly inhibit ATP-dependent proton accumulation in the vesicles without inhibiting ATP hydrolysis. This uncoupling effect of the fatty acids could be abolished by the addition of fatty acid-free bovine serum albumin (BSA). Similarly, the proton transport capacity of ageing vesicles could be restored by addition of BSA. The latter findings may explain why isolated plasma membranes so often exhibit increased permeability to protons on ageing.

  6. Development of an Efficient Protein Extraction Method Compatible with LC-MS/MS for Proteome Mapping in Two Australian Seagrasses Zostera muelleri and Posidonia australis

    PubMed Central

    Jiang, Zhijian; Kumar, Manoj; Padula, Matthew P.; Pernice, Mathieu; Kahlke, Tim; Kim, Mikael; Ralph, Peter J.

    2017-01-01

    The availability of the first complete genome sequence of the marine flowering plant Zostera marina (commonly known as seagrass) in early 2016, is expected to significantly raise the impact of seagrass proteomics. Seagrasses are marine ecosystem engineers that are currently declining worldwide at an alarming rate due to both natural and anthropogenic disturbances. Seagrasses (especially species of the genus Zostera) are compromised for proteomic studies primarily due to the lack of efficient protein extraction methods because of their recalcitrant cell wall which is rich in complex polysaccharides and a high abundance of secondary metabolites in their cells. In the present study, three protein extraction methods that are commonly used in plant proteomics i.e., phenol (P); trichloroacetic acid/acetone/SDS/phenol (TASP); and borax/polyvinyl-polypyrrolidone/phenol (BPP) extraction, were evaluated quantitatively and qualitatively based on two dimensional isoelectric focusing (2D-IEF) maps and LC-MS/MS analysis using the two most abundant Australian seagrass species, namely Zostera muelleri and Posidonia australis. All three tested methods produced high quality protein extracts with excellent 2D-IEF maps in P. australis. However, the BPP method produces better results in Z. muelleri compared to TASP and P. Therefore, we further modified the BPP method (M-BPP) by homogenizing the tissue in a modified protein extraction buffer containing both ionic and non-ionic detergents (0.5% SDS; 1.5% Triton X-100), 2% PVPP and protease inhibitors. Further, the extracted proteins were solubilized in 0.5% of zwitterionic detergent (C7BzO) instead of 4% CHAPS. This slight modification to the BPP method resulted in a higher protein yield, and good quality 2-DE maps with a higher number of protein spots in both the tested seagrasses. Further, the M-BPP method was successfully utilized in western-blot analysis of phosphoenolpyruvate carboxylase (PEPC—a key enzyme for carbon metabolism

  7. Functionalizable and ultra stable nanoparticles coated with zwitterionic poly(carboxybetaine) in undiluted blood serum.

    PubMed

    Yang, Wei; Zhang, Lei; Wang, Shanlin; White, Andrew D; Jiang, Shaoyi

    2009-10-01

    A new surface chemistry presenting an abundance of functional groups for ligand immobilization in an ultra-low fouling background all in one material for nanoparticles was introduced. This surface platform, as demonstrated by zwitterionic poly(carboxybetaine acrylamide) (polyCBAA) coated nanoparticles, is not only ultra stable in undiluted human blood serum, but also can be conjugated to biomolecules conveniently and effectively. Thus, this surface chemistry is ideal to create multi-functional nanoparticles for targeted delivery and diagnostics. In addition, this work clearly shows that 10% blood serum commonly used to evaluate the stability of nanoparticles is insufficient and a new evaluation criterion with undiluted blood serum is recommended.

  8. Production of lipase and protease from an indigenous Pseudomonas aeruginosa strain and their evaluation as detergent additives: compatibility study with detergent ingredients and washing performance.

    PubMed

    Grbavčić, Sanja; Bezbradica, Dejan; Izrael-Živković, Lidija; Avramović, Nataša; Milosavić, Nenad; Karadžić, Ivanka; Knežević-Jugović, Zorica

    2011-12-01

    An indigenous Pseudomonas aeruginosa strain has been studied for lipase and protease activities for their potential application in detergents. Produced enzymes were investigated in order to assess their compatibility with several surfactants, oxidizing agents and commercial detergents. The crude lipase appeared to retain high activity and stability in the presence of several surfactants and oxidizing agents and it was insusceptible to proteolysis. Lutensol® XP80 and Triton® X-100 strongly activated the lipase for a long period (up to 40 and 30% against the control after 1h) while the protease activity was enhanced by the addition of Triton® WR1339 and Tween® 80. The washing performance of the investigated surfactants was significantly improved with the addition of the crude enzyme preparation. Studies were further undertaken to improve enzymes production. The optimization of fermentation conditions led to an 8-fold increase of lipase production, while the production of protease was enhanced by 60%. Copyright © 2011 Elsevier Ltd. All rights reserved.

  9. Fourier transform infrared spectroscopic study of the interactions of a strongly antimicrobial but weakly hemolytic analogue of gramicidin S with lipid micelles and lipid bilayer membranes.

    PubMed

    Lewis, Ruthven N A H; Kiricsi, Monika; Prenner, Elmar J; Hodges, Robert S; McElhaney, Ronald N

    2003-01-21

    Cyclo[VKLdKVdYPLKVKLdYP] (GS14dK(4)), a synthetic tetradecameric ring-size analogue of the naturally occurring antimicrobial peptide gramicidin S (GS), retains the strong antimicrobial activity of GS but is 15-20 times less hemolytic. To characterize its interaction with lipid membranes and to understand the molecular basis of its capacity to lyse bacterial cells, in preference to erythrocytes, we have investigated the interactions of GS14dK(4) with detergent micelles and with lipid bilayer model membranes by Fourier transform infrared spectroscopy and compared our results with those of a similar study of GS [Lewis, R. N. A. H., et al. (1999) Biochemistry 38, 15193-15203]. In both aqueous and organic solvent solutions, GS14dK(4) adopts a beta-sheet conformation that is somewhat distorted and more sensitive to the polarity of its environment than GS. Like GS, GS14dK(4) is completely or partially excluded from gel-state lipid bilayers but interacts strongly with liquid-crystalline lipid bilayers and detergent micelle, and interacts more strongly with more fluid liquid-crystalline lipid systems. However, its interactions are more strongly influenced by membrane lipid order and fluidity, and unlike GS, it is essentially excluded from cholesterol-containing phospholipid bilayers. Also, GS14dK(4) is excluded from cationic lipid bilayers, but partitions more strongly and/or penetrates more deeply into anionic lipid bilayers than into those composed of either zwitterionic or nonionic lipids. Anionic lipids also facilitate GS14dK(4) interactions with multicomponent lipid bilayers which are predominantly zwitterionic or nonionic. Although GS14dK(4) generally penetrates and/or partitions into zwitterionic or uncharged lipid bilayers less strongly than does GS, its greater size and altered distribution of positive charges make it intrinsically more perturbing with regard to membrane organization once associated with lipid bilayers. This fact, combined with its relatively

  10. Calibration transfer of a Raman spectroscopic quantification method from at-line to in-line assessment of liquid detergent compositions.

    PubMed

    Brouckaert, D; Uyttersprot, J-S; Broeckx, W; De Beer, T

    2017-06-08

    The industrial production of liquid detergent compositions entails delicate balance of ingredients and process steps. In order to assure high quality and productivity in the manufacturing line, process analytical technology tools such as Raman spectroscopy are to be implemented. Marked chemical specificity, negligible water interference and high robustness are ascribed to this process analytical technique. Previously, at-line calibration models have been developed for determining the concentration levels of the being studied liquid detergents main ingredients from Raman spectra. A strategy is now proposed to transfer such at-line developed regression models to an in-line set-up, allowing real-time dosing control of the liquid detergent composition under production. To mimic in-line manufacturing conditions, liquid detergent compositions are created in a five-liter vessel with an overhead mixer. Raman spectra are continuously acquired by pumping the detergent under production via plastic tubing towards a Raman superhead probe, which is incorporated into a metal frame with a sapphire window facing the detergent fluid. Two at-line developed partial least squares (PLS) models are aimed at transferring, predicting the concentration of surfactant 1 and polymer 2 in the examined liquid detergent composition. A univariate slope/bias correction (SBC) is investigated, next to three well-acknowledged multivariate transformation methods: direct, piecewise and double-window piecewise direct standardization. Transfer is considered successful when the magnitude of the validation sets root mean square error of prediction (RMSEP) is similar to or smaller than the corresponding at-line prediction error. The transferred model offering the most promising outcome is further subjected to an exhaustive statistical evaluation, in order to appraise the applicability of the suggested calibration transfer method. Interval hypothesis tests are thereby performed for method comparison. It is

  11. Rat brain gamma-secretase activity is highly influenced by detergents.

    PubMed

    Frånberg, Jenny; Welander, Hedvig; Aoki, Mikio; Winblad, Bengt; Tjernberg, Lars O; Frykman, Susanne

    2007-06-26

    Gamma-secretase is important for the development of Alzheimer's disease, since it is a crucial enzyme for the generation of the pathogenic amyloid beta-peptide (Abeta). Most data on gamma-secretase is derived from studies in cell lines overexpressing gamma-secretase components or amyloid precursor protein (APP), and since gamma-secretase is a transmembrane protein complex, detergents have been frequently used to facilitate the studies. However, no extensive comparison of the influence of different detergents at different concentrations on gamma-secretase activity in preparations from brain has been made. Here, we establish the optimal conditions for gamma-secretase activity in rat brain, using an activity assay detecting endogenous production of the APP intracellular domain, which is generated when gamma-secretase cleaves the APP C-terminal fragments. We performed a subcellular fractionation and noted the highest gamma-secretase activity in the 100000g pellet and that the optimal pH was around 7. We found that gamma-secretase was active for at least 16 h at 37 degrees C and that the endogenous substrate levels were sufficient for activity measurements. The highest activity was obtained in 0.4% CHAPSO, which is slightly below the critical micelle concentration (0.5%) for this detergent, but the complex was not solubilized efficiently at this concentration. On the other hand, 1% CHAPSO solubilized a substantial amount of the gamma-secretase components, but the activity was low. The activity was fully restored by diluting the sample to 0.4% CHAPSO. Therefore, using 1% CHAPSO for solubilization and subsequently diluting the sample to 0.4% is an appropriate procedure for obtaining a soluble, highly active gamma-secretase from rat brain.

  12. Fluorophore Absorption Size Exclusion Chromatography (FA-SEC): An Alternative Method for High-Throughput Detergent Screening of Membrane Proteins.

    PubMed

    Lin, Sung-Yao; Sun, Xing-Han; Hsiao, Yu-Hsuan; Chang, Shao-En; Li, Guan-Syun; Hu, Nien-Jen

    2016-01-01

    Membrane proteins play key roles in many fundamental functions in cells including ATP synthesis, ion and molecule transporter, cell signalling and enzymatic reactions, accounting for ~30% genes of whole genomes. However, the hydrophobic nature of membrane proteins frequently hampers the progress of structure determination. Detergent screening is the critical step in obtaining stable detergent-solubilized membrane proteins and well-diffracting protein crystals. Fluorescence Detection Size Exclusion Chromatography (FSEC) has been developed to monitor the extraction efficiency and monodispersity of membrane proteins in detergent micelles. By tracing the FSEC profiles of GFP-fused membrane proteins, this method significantly enhances the throughput of detergent screening. However, current methods to acquire FSEC profiles require either an in-line fluorescence detector with the SEC equipment or an off-line spectrofluorometer microplate reader. Here, we introduce an alternative method detecting the absorption of GFP (FA-SEC) at 485 nm, thus making this methodology possible on conventional SEC equipment through the in-line absorbance spectrometer. The results demonstrate that absorption is in great correlation with fluorescence of GFP. The comparably weaker absorption signal can be improved by using a longer path-length flow cell. The FA-SEC profiles were congruent with the ones plotted by FSEC, suggesting FA-SEC could be a comparable and economical setup for detergent screening of membrane proteins.

  13. [An experimental evaluation of the efficacy of 4 types of detergents for 3 types of dyes to which workers in color-printing plants are exposed].

    PubMed

    Terzaghi, G F; Settimi, L; Peverelli, C; Sevosi, L; Duca, P G

    1996-01-01

    The efficacy of 4 commercial cleansing products was tested with 3 colouring agents widely used in the dyeing industry in a randomised double blind trail involving 8 workers each time. A between-detergents statistically significant difference was observed; the interaction (detergents x colouring agents) was significant. The efficacy of type A detergent was higher for type I-III dyers, while the efficacy of type C detergent, which widely used was lowest.

  14. Glycosaminoglycan-resistant and pH-sensitive lipid-coated DNA complexes produced by detergent removal method.

    PubMed

    Lehtinen, Julia; Hyvönen, Zanna; Subrizi, Astrid; Bunjes, Heike; Urtti, Arto

    2008-10-21

    Cationic polymers are efficient gene delivery vectors in in vitro conditions, but these carriers can fail in vivo due to interactions with extracellular polyanions, i.e. glycosaminoglycans (GAG). The aim of this study was to develop a stable gene delivery vector that is activated at the acidic endosomal pH. Cationic DNA/PEI complexes were coated by 1,2-dioleylphosphatidylethanolamine (DOPE) and cholesteryl hemisuccinate (CHEMS) (3:2 mol/mol) using two coating methods: detergent removal and mixing with liposomes prepared by ethanol injection. Only detergent removal produced lipid-coated DNA complexes that were stable against GAGs, but were membrane active at low pH towards endosome mimicking liposomes. In relation to the low cellular uptake of the coated complexes, their transfection efficacy was relatively high. PEGylation of the coated complexes increased their cellular uptake but reduced the pH-sensitivity. Detergent removal was thus a superior method for the production of stable, but acid activatable, lipid-coated DNA complexes.

  15. Restricting detergent protease action to surface of protein fibres by chemical modification.

    PubMed

    Schroeder, M; Lenting, H B M; Kandelbauer, A; Silva, C J S M; Cavaco-Paulo, A; Gübitz, G M

    2006-10-01

    Due to their excellent properties, such as thermostability, activity over a broad range of pH and efficient stain removal, proteases from Bacillus sp. are commonly used in the textile industry including industrial processes and laundry and represent one of the most important groups of enzymes. However, due to the action of proteases, severe damage on natural protein fibres such as silk and wool result after washing with detergents containing proteases. To include the benefits of proteases in a wool fibre friendly detergent formulation, the soluble polymer polyethylene glycol (PEG) was covalently attached to a protease from Bacillus licheniformis. In contrast to activation of PEG with cyanuric chloride (50%) activation with 1,1'-carbonyldiimidazole (CDI) lead to activity recovery above 90%. With these modified enzymes, hydrolytic attack on wool fibres could be successfully prevented up to 95% compared to the native enzymes. Colour difference (DeltaE) measured in the three dimensional colour space showed good stain removal properties for the modified enzymes. Furthermore, half-life of the modified enzymes in buffers and commercial detergents solutions was nearly twice as high as those of the non-modified enzymes with values of up to 63 min. Out of the different modified proteases especially the B. licheniformis protease with the 2.0-kDa polymer attached both retained stain removal properties and did not hydrolyse/damage wool fibres.

  16. Poly(zwitterionic)protein conjugates offer increased stability without sacrificing binding affinity or bioactivity

    NASA Astrophysics Data System (ADS)

    Keefe, Andrew J.; Jiang, Shaoyi

    2012-01-01

    Treatment with therapeutic proteins is an attractive approach to targeting a number of challenging diseases. Unfortunately, the native proteins themselves are often unstable in physiological conditions, reducing bioavailability and therefore increasing the dose that is required. Conjugation with poly(ethylene glycol) (PEG) is often used to increase stability, but this has a detrimental effect on bioactivity. Here, we introduce conjugation with zwitterionic polymers such as poly(carboxybetaine). We show that poly(carboxybetaine) conjugation improves stability in a manner similar to PEGylation, but that the new conjugates retain or even improve the binding affinity as a result of enhanced protein-substrate hydrophobic interactions. This chemistry opens a new avenue for the development of protein therapeutics by avoiding the need to compromise between stability and affinity.

  17. Leptospira santorosai Serovar Shermani detergent extract induces an increase in fibronectin production through a Toll-like receptor 2-mediated pathway.

    PubMed

    Tian, Ya-Chung; Hung, Cheng-Chieh; Li, Yi-Jung; Chen, Yung-Chang; Chang, Ming-Yang; Yen, Tzung-Hai; Hsu, Hsiang-Hao; Wu, Mai-Szu; Phillips, Aled; Yang, Chih-Wei

    2011-03-01

    Leptospirosis can activate inflammatory responses through Toll-like receptors (TLRs) and may cause renal tubulointerstitial fibrosis characterized by the accumulation of extracellular matrix (ECM). We have previously demonstrated that Leptospira santorosai serovar Shermani detergent extract stimulates ECM accumulation in vitro. The aim of this study was to examine the mechanistic basis of these previous observations and, in particular, to examine the potential involvement of TLRs. The addition of serovar Shermani detergent extract led to an increase in fibronectin gene expression and production. Inhibition of TLR2 but not TLR4 expression abrogated serovar Shermani detergent extract-mediated increases in fibronectin production. This response was also blocked by the knockdown of the gene expression of the TLR2 downstream transducers myeloid differentiation factor 88 (MyD88) and tumor necrosis factor receptor-associated factor 6 (TRAF6). Serovar Shermani detergent extract also activated nuclear factor-κB, and its inhibition by curcumin-attenuated serovar Shermani detergent extract induced increases in fibronectin production. These effects were also mimicked by the specific TLR2 agonist, Pam(3)CsK(4), a response that was also abrogated by the knockdown of MyD88 and TRAF6. Similarly, the administration of live leptospires to cells also induced fibronectin production that was blocked by inhibition of TLR2 and MyD88 expression. In conclusion, serovar Shermani detergent extract can induce fibronectin production through the TLR2-associated cascade, providing evidence of an association between TLRs and leptospirosis-mediated ECM deposition.

  18. Trends in Thermostability Provide Information on the Nature of Substrate, Inhibitor, and Lipid Interactions with Mitochondrial Carriers*

    PubMed Central

    Crichton, Paul G.; Lee, Yang; Ruprecht, Jonathan J.; Cerson, Elizabeth; Thangaratnarajah, Chancievan; King, Martin S.; Kunji, Edmund R. S.

    2015-01-01

    Mitochondrial carriers, including uncoupling proteins, are unstable in detergents, which hampers structural and mechanistic studies. To investigate carrier stability, we have purified ligand-free carriers and assessed their stability with a fluorescence-based thermostability assay that monitors protein unfolding with a thiol-reactive dye. We find that mitochondrial carriers from both mesophilic and thermophilic organisms exhibit poor stability in mild detergents, indicating that instability is inherent to the protein family. Trends in the thermostability of yeast ADP/ATP carrier AAC2 and ovine uncoupling protein UCP1 allow optimal conditions for stability in detergents to be established but also provide mechanistic insights into the interactions of lipids, substrates, and inhibitors with these proteins. Both proteins exhibit similar stability profiles across various detergents, where stability increases with the size of the associated detergent micelle. Detailed analysis shows that lipids stabilize carriers indirectly by increasing the associated detergent micelle size, but cardiolipin stabilizes by direct interactions as well. Cardiolipin reverses destabilizing effects of ADP and bongkrekic acid on AAC2 and enhances large stabilizing effects of carboxyatractyloside, revealing that this lipid interacts in the m-state and possibly other states of the transport cycle, despite being in a dynamic interface. Fatty acid activators destabilize UCP1 in a similar way, which can also be prevented by cardiolipin, indicating that they interact like transport substrates. Our controls show that carriers can be soluble but unfolded in some commonly used detergents, such as the zwitterionic Fos-choline-12, which emphasizes the need for simple validation assays like the one used here. PMID:25653283

  19. A Lab Experience to Illustrate the Physicochemical Principles of Detergency

    ERIC Educational Resources Information Center

    Poce-Fatou, J. A.; Bethencourt-Nunez, M.; Moreno, C.; Pinto-Ganfornina, J. J.; Moreno-Dorado, F. J.

    2008-01-01

    This article presents a lab experience to study detergency from a physicochemical point of view intended for undergraduate students. By means of a simple experimental device, we analyze the influence of the surfactant concentration in both distilled water and tap water. Our method is based on the measurement of diffuse reflectances of polyester…

  20. Synthesis and solution aggregation studies of a suite of mixed neutral and zwitterionic chromophores for second-order nonlinear optics.

    PubMed

    Peddie, Victoria; Anderson, Jack; Harvey, Joanne E; Smith, Gerald J; Kay, Andrew

    2014-11-07

    We report details of the synthesis of a series of bi- and trichromophores. These compounds contain mixtures of chromophores that have zwitterionic (ZWI) and neutral ground state (NGS) components covalently attached to each other. The neutral ground state moieties are based on dyes with aniline donors--such as Disperse Red 1--whereas the zwitterionic components are derived from chromophores with pro-aromatic donors such as 1,4-dihydropyridinylidene. By combining both ZWI and NGS components, we aim to develop novel compounds for nonlinear optics in which there is an enhancement of the overall hyperpolarizability coupled with a decrease in the net dipole moment. Thus, this approach should eliminate the electrostatic effects that result when only one type of chromophore is used, and so reduce the likelihood of undesirable aggregation occurring. This, in turn, should enable us to realize organic materials with large macroscopic optical nonlinearities. An analysis of the UV-vis results suggests that there is a strong dependence on solvent polarity that determines whether the embedded constituents should be treated as discrete elements; in low polarity solvents, there appear to be strong intramolecular interactions occurring, particularly when a 1,4-quinolinylidene-based donor is used in the ZWI component.

  1. [Membrane lipids and electron transfer. Effects of four detergents on NADH-ferricyanide reductase and NADH-cytochrome c reductase activities of potato tuber microsomes].

    PubMed

    Jolliot, A; Mazliak, P

    1977-10-17

    The NADH-ferricyanure reductase activity of Potato microsomes is stimulated by non ionic detergents (Triton X100 and Tween80) and is partially inhibited by ionic detergents (sodium-cholate and deoxycholate). All these four detergents progressively decreased the NADH-cytochrome c reductase in the following order: sodium deoxycholate greater than Triton X100 greater than sodium cholate greater than Tween80.

  2. Dynamic Interaction of cBid with Detergents, Liposomes and Mitochondria

    PubMed Central

    Bleicken, Stephanie; García-Sáez, Ana J.; Conte, Elena; Bordignon, Enrica

    2012-01-01

    The BH3-only protein Bid plays a key role in the induction of mitochondrial apoptosis, but its mechanism of action is still not completely understood. Here we studied the two main activation events of Bid: Caspase-8 cleavage and interaction with the membrane bilayer. We found a striking reversible behaviour of the dissociation-association events between the Bid fragments p15 and p7. Caspase-8 cleavage does not induce per se separation of the two Bid fragments, which remain in a stable complex resembling the full length Bid. Detergents trigger a complete dissociation, which can be fully reversed by detergent removal in a range of protein concentrations from 100 µM down to 500 nM. Incubation of cBid with cardiolipin-containing liposomes leads to partial dissociation of the complex. Only p15 (tBid) fragments are found at the membrane, while p7 shows no tendency to interact with the bilayer, but complete removal of p7 strongly increases the propensity of tBid to become membrane-associated. Despite the striking structural similarities of inactive Bid and Bax, Bid does not form oligomers and reacts differently in the presence of detergents and membranes, highlighting clear differences in the modes of action of the two proteins. The partial dissociation of cBid triggered by the membrane is suggested to depend on the strong and specific interaction between p15 and p7. The reversible disassembly and re-assembly of the cBid molecules at the membrane was as well proven by EPR using spin labeled cBid in the presence of isolated mitochondria. The observed dynamic dissociation of the two Bid fragments could allow the assistance to the pore-forming Bax to occur repeatedly and may explain the proposed “hit-and-run" mode of action of Bid at the bilayer. PMID:22540011

  3. Ionic-liquid-induced changes in the properties of aqueous zwitterionic surfactant solution: solvent and rotational relaxation studies.

    PubMed

    Rao, Vishal Govind; Ghatak, Chiranjib; Ghosh, Surajit; Mandal, Sarthak; Sarkar, Nilmoni

    2012-03-29

    In the recent past, the chameleon-like nature of zwitterionic micelles has been utilized for performing electrophilic, nucleophilic, base, and acid catalyzed reactions. But the use of simple salts to induce the zwitterionic character limits the variation to inorganic cations and anions only. To overcome this problem, we have used room temperature ionic liquids (RTILs), which can be tailored according to need. More precisely, we have shown the effect of added RTILs on the nature of water molecules in the palisade layer of a zwitterionic (N-hexadecyl-N,N-dimethylammonio-1-propanesulfonate (SB-16)) micelle using solvation and rotational relaxation studies of C-153 dye. We have carried out a comparative study of changes in the solvent and rotational relaxation parameters of C-153 in an aqueous solution of SB-16 upon addition of three different ionic liquids (ILs): 1-ethyl-3-methylimidazolium ethyl sulfate [C(2)mim][C(2)SO(4)], 1-ethyl-3-methylimidazolium n-butyl sulfate [C(2)mim][C(4)SO(4)], and 1-ethyl-3-methylimidazolium n-hexyl sulfate [C(2)mim][C(6)SO(4)]. It has been observed that in the presence of added RTILs the solvation dynamics become faster and the change in solvation dynamics is more pronounced in the case of [C(2)mim][C(6)SO(4)] compared to that for [C(2)mim][C(4)SO(4)] and [C(2)mim][C(2)SO(4)]. This can be accounted for by considering the increased water penetration (increased microfluidity) with the addition of ILs. In accordance with solvation dynamics results, fluorescence anisotropy studies also indicate an increase in microfluidity of the palisade layer of the SB-16 micelle with the added RTILs. The average rotational relaxation time in 28 mM SB-16 was found to be 1.12 ns. With the addition of 800 mM [C(2)mim][C(2)SO(4)], the average rotational relaxation time remains the same (1.12 ns), whereas with the addition of 800 mM [C(2)mim][C(6)SO(4)] it decreases to 0.40 ns. This observation is in agreement with our earlier report on the microfluidity of

  4. Optimized formation of detergent micelles of beta-carotene and retinal production using recombinant human beta,beta-carotene 15,15'-monooxygenase.

    PubMed

    Kim, Nam-Hee; Kim, Yeong-Su; Kim, Hye-Jung; Oh, Deok-Kun

    2008-01-01

    The formation of beta-carotene detergent micelles and their conversion into retinal by recombinant human beta,beta-carotene 15,15'-monooxygenase was optimized under aqueous conditions. Toluene was the most hydrophobic among the organic solvents tested; thus, it was used to dissolve beta-carotene, which is a hydrophobic compound. Tween 80 was selected as the detergent because it supported the highest level of retinal production among all of the detergents tested. The maximum production of retinal was achieved in detergent micelles containing 200 mg/L of beta-carotene and 2.4% (w/v) Tween 80. Under these conditions, the recombinant enzyme produced 97 mg/L of retinal after 16 h with a conversion yield of 48.5% (w/w). The amount of retinal produced, which is the highest ever reported, is a result of the ability of our system to dissolve large amounts of beta-carotene.

  5. Suppressing surface reconstruction of superhydrophobic PDMS using a superhydrophilic zwitterionic polymer.

    PubMed

    Keefe, Andrew J; Brault, Norman D; Jiang, Shaoyi

    2012-05-14

    Poly(dimethyl siloxane) (PDMS) is extensively used for biomedical applications due to its low cost, ease of fabrication, high durability and flexibility, oxygen permeability, and self-healing properties. PDMS, however, has some significant drawbacks. PDMS endures unacceptably high levels of nonspecific protein fouling when used with biological samples due to its superhydrophobic characteristics. Unfortunately, conventional surface modification methods do not work for PDMS due to its low glass transition temperature. This phenomenon has been well-known for years as "hydrophobic regeneration". For the same reason, it is also very difficult to bring functionalities onto PDMS surfaces. Herein, we demonstrate how a superhydrophilic zwitterionic material, poly(carboxybetaine methacrylate) (pCBMA), can provide a highly stable coating with long-term stabilty due to the sharp contrast in hydrophobicity between pCBMA and PDMS. This material is able to suppress nonspecific protein adsorption in complex media and functionalize desired biomolecules needed in applications, such as diagnostics, without sacrificing its nonfouling characteristics.

  6. Suppressing Surface Reconstruction of Superhydrophobic PDMS using a Superhydrophilic Zwitterionic Polymer

    PubMed Central

    Keefe, Andrew J.; Brault, Norman D.; Jiang, Shaoyi

    2014-01-01

    Poly(dimethyl siloxane) (PDMS) is extensively used for biomedical applications due to its low cost, ease of fabrication, high durability and flexibility, oxygen permeability, and self-healing properties. PDMS, however, has some significant drawbacks. PDMS endures unacceptably high levels of non-specific protein fouling when used with biological samples due to its superhydrophobic characteristics. Unfortunately, conventional surface modification methods do not work for PDMS due to its low glass transition temperature. This phenomenon has been well-known for years as “hydrophobic regeneration”. For the same reason, it is also very difficult to bring functionalities onto PDMS surfaces. Herein, we demonstrate how a superhydrophilic zwitterionic material, poly(carboxybetaine methacrylate) (pCBMA), can provide a highly stable coating with long term stabilty due to the sharp contrast in hydrophobicity between pCBMA and PDMS. This material is able to suppress nonspecific protein adsorption in complex media and functionalize desired biomolecules needed in applications, such as diagnostics, without sacrificing its nonfouling characteristics. PMID:22512660

  7. A comparative spectroscopic and kinetic study of photoexcitations in detergent-isolated and membrane-embedded LH2 light-harvesting complexes.

    PubMed

    Freiberg, Arvi; Rätsep, Margus; Timpmann, Kõu

    2012-08-01

    Integral membrane proteins constitute more than third of the total number of proteins present in organisms. Solubilization with mild detergents is a common technique to study the structure, dynamics, and catalytic activity of these proteins in purified form. However beneficial the use of detergents may be for protein extraction, the membrane proteins are often denatured by detergent solubilization as a result of native lipid membrane interactions having been modified. Versatile investigations of the properties of membrane-embedded and detergent-isolated proteins are, therefore, required to evaluate the consequences of the solubilization procedure. Herein, the spectroscopic and kinetic fingerprints have been established that distinguish excitons in individual detergent-solubilized LH2 light-harvesting pigment-protein complexes from them in the membrane-embedded complexes of purple photosynthetic bacteria Rhodobacter sphaeroides. A wide arsenal of spectroscopic techniques in visible optical range that include conventional broadband absorption-fluorescence, fluorescence anisotropy excitation, spectrally selective hole burning and fluorescence line-narrowing, and transient absorption-fluorescence have been applied over broad temperature range between physiological and liquid He temperatures. Significant changes in energetics and dynamics of the antenna excitons upon self-assembly of the proteins into intracytoplasmic membranes are observed, analyzed, and discussed. This article is part of a Special Issue entitled: Photosynthesis Research for Sustainability: from Natural to Artificial. Copyright © 2011. Published by Elsevier B.V.

  8. The structure of ions and zwitterionic lipids regulates the charge of dipolar membranes.

    PubMed

    Szekely, Or; Steiner, Ariel; Szekely, Pablo; Amit, Einav; Asor, Roi; Tamburu, Carmen; Raviv, Uri

    2011-06-21

    In pure water, zwitterionic lipids form lamellar phases with an equilibrium water gap on the order of 2 to 3 nm as a result of the dominating van der Waals attraction between dipolar bilayers. Monovalent ions can swell those neutral lamellae by a small amount. Divalent ions can adsorb onto dipolar membranes and charge them. Using solution X-ray scattering, we studied how the structure of ions and zwitterionic lipids regulates the charge of dipolar membranes. We found that unlike monovalent ions that weakly interact with all of the examined dipolar membranes, divalent and trivalent ions adsorb onto membranes containing lipids with saturated tails, with an association constant on the order of ∼10 M(-1). One double bond in the lipid tail is sufficient to prevent divalent ion adsorption. We suggest that this behavior is due to the relatively loose packing of lipids with unsaturated tails that increases the area per lipid headgroup, enabling their free rotation. Divalent ion adsorption links two lipids and limits their free rotation. The ion-dipole interaction gained by the adsorption of the ions onto unsaturated membranes is insufficient to compensate for the loss of headgroup free-rotational entropy. The ion-dipole interaction is stronger for cations with a higher valence. Nevertheless, polyamines behave as monovalent ions near dipolar interfaces in the sense that they interact weakly with the membrane surface, whereas in the bulk their behavior is similar to that of multivalent cations. Advanced data analysis and comparison with theory provide insight into the structure and interactions between ion-induced regulated charged interfaces. This study models biologically relevant interactions between cell membranes and various ions and the manner in which the lipid structure governs those interactions. The ability to monitor these interactions creates a tool for probing systems that are more complex and forms the basis for controlling the interactions between dipolar

  9. Antifouling Properties of a Self-Assembling Glutamic Acid-Lysine Zwitterionic Polymer Surface Coating.

    PubMed

    Ziemba, Christopher; Khavkin, Maria; Priftis, Dimitris; Acar, Handan; Mao, Jun; Benami, Maya; Gottlieb, Moshe; Tirrell, Matthew; Kaufman, Yair; Herzberg, Moshe

    2018-04-23

    There is a need for the development of antifouling materials to resist adsorption of biomacromolecules. Here we describe the preparation of a novel zwitterionic block copolymer with the potential to prevent or delay the formation of microbial biofilms. The block copolymer comprised a zwitterionic (hydrophilic) section of alternating glutamic acid (negatively charged) and lysine (positively charged) units and a hydrophobic polystyrene section. Cryo-TEM and dynamic-light-scattering (DLS) results showed that, on average, the block copolymer self-assembled into 7-nm-diameter micelles in aqueous solutions (0 to 100 mM NaCl, pH 6). Quartz crystal microbalance with dissipation monitoring (QCM-D), atomic force microscopy (AFM), and contact angle measurements demonstrated that the block copolymer self-assembled into a brush-like monolayer on polystyrene surfaces. The brush-like monolayer produced from a 100 mg/L block copolymer solution exhibited an average distance, d, of approximately 4-8 nm between each block copolymer molecule (center to center). Once the brush-like monolayer self-assembled, it reduced EPS adsorption onto the polystyrene surface by ∼70% (mass), reduced the rate of bacterial attachment by >80%, and inhibited the development of thick biofilms. QCM-D results revealed that the EPS molecules penetrate between the chains of the brush and adsorb onto the polystyrene surface. Additionally, AFM analyses showed that the brush-like monolayer prevents the adhesion of large (> d) hydrophilic colloids onto the surface via hydration repulsion; however, molecules or colloids small enough to fit between the brush polymers (< d) were able to be adsorbed onto the surface via van der Waals interactions. Overall, we found that the penetration of extracellular organelles, as well as biopolymers through the brush, is critical for the failure of the antifouling coating, and likely could be prevented through tuning of the brush density. Stability and biofilm development

  10. Cleanser, Detergent, Personal Care Product Pretreatment Evaluation

    NASA Technical Reports Server (NTRS)

    Adam, Niklas

    2010-01-01

    The purpose of the Cleanser, Detergent, Personal Care Product, and Pretreatment Evaluation & Selection task is to identify the optimal combination of personal hygiene products, crew activities, and pretreatment strategies to provide the crew with sustainable life support practices and a comfortable habitat. Minimal energy, mass, and crew time inputs are desired to recycle wastewater during long duration missions. This document will provide a brief background on the work this past year supporting the ELS Distillation Comparison Test, issues regarding use of the hygiene products originally chosen for the test, methods and results used to select alternative products, and lessons learned from testing.

  11. Thermodynamic Analysis of the Selectivity Enhancement Obtained by Using Smart Hydrogels That Are Zwitterionic When Detecting Glucose With Boronic Acid Moieties

    PubMed Central

    Horkay, F.; Cho, S. H.; Tathireddy, P.; Rieth, L.; Solzbacher, F.; Magda, J.

    2011-01-01

    Because the boronic acid moiety reversibly binds to sugar molecules and has low cytotoxicity, boronic acid-containing hydrogels are being used in a variety of implantable glucose sensors under development, including sensors based on optical, fluorescence, and swelling pressure measurements. However, some method of glucose selectivity enhancement is often necessary, because isolated boronic acid molecules have a binding constant with glucose that is some forty times smaller than their binding constant with fructose, the second most abundant sugar in the human body. In many cases, glucose selectivity enhancement is obtained by incorporating pendant tertiary amines into the hydrogel network, thereby giving rise to a hydrogel that is zwitterionic at physiological pH. However, the mechanism by which incorporation of tertiary amines confers selectivity enhancement is poorly understood. In order to clarify this mechanism, we use the osmotic deswelling technique to compare the thermodynamic interactions of glucose and fructose with a zwitterionic smart hydrogel containing boronic acid moieties. We also investigate the change in the structure of the hydrogel that occurs when it binds to glucose or to fructose using the technique of small angle neutron scattering. PMID:22190765

  12. Lidocaine effect on flotillin-2 distribution in detergent-resistant membranes of equine jejunal smooth muscle in vitro.

    PubMed

    Tappenbeck, Karen; Schmidt, Sonja; Feige, Karsten; Naim, Hassan Y; Huber, Korinna

    2014-05-01

    Lidocaine is the most commonly chosen prokinetic for treating postoperative ileus in horses, a motility disorder associated with ischaemia-reperfusion injury of intestinal tissues. Despite the frequent use of lidocaine, the mechanism underlying its prokinetic effects is still unclear. Previous studies suggested that lidocaine altered cell membrane characteristics of smooth muscle cells. Therefore, the present study aimed to elucidate effects of lidocaine administration on characteristics of detergent-resistant membranes in equine jejunal smooth muscle. Lidocaine administration caused significant redistribution of flotillin-2, a protein marker of detergent-resistant membranes, in fractions of sucrose-density-gradients obtained from ischaemia-reperfusion injured smooth muscle solubilised with Triton X-100. It was concluded that lidocaine induced disruption of detergent-resistant membranes which might affect ion channel activity and therefore enhance smooth muscle contractility. Copyright © 2014 Elsevier Ltd. All rights reserved.

  13. Relevance of sensitization to occupational allergy and asthma in the detergent industry.

    PubMed

    Basketter, David; Berg, Ninna; Kruszewski, Francis H; Sarlo, Katherine; Concoby, Beth

    2012-01-01

    There exists considerable historic experience of the relationship between exposure and both the induction of sensitization and the elicitation of respiratory symptoms from industrial enzymes of bacterial and fungal origin used in a wide variety of detergent products. The detergent industry in particular has substantial experience of how the control of exposure leads to limitation of sensitization with low risk of symptoms. However, the experience also shows that there are substantial gaps in knowledge, even when the potential occupational allergy problem is firmly under control, and also that the relationship between exposure and sensitization can be hard to establish. The latter aspect includes a poor appreciation of how peak exposures and low levels of exposure over time contribute to sensitization. Furthermore, while a minority of workers develop specific IgE, essentially none appear to have symptoms, a situation which appears to contradict the allergy dogma that, once sensitized, an individual will react to much lower levels of exposure. For enzymes, the expression of symptoms occurs at similar or higher levels than those that cause induction. In spite of some knowledge gaps, medical surveillance programs and constant air monitoring provide the tools for successful management of enzymes in the occupational setting. Ultimately, the knowledge gained from the occupational setting facilitates the completion of safety assessments for consumer exposure to detergent enzymes. Such assessments have been proven to be correct by the decades of safe use both occupationally and in consumer products.

  14. Spectroscopic studies on the active site of hydroperoxide lyase; the influence of detergents on its conformation.

    PubMed

    Noordermeer, M A; Veldink, G A; Vliegenthart, J F

    2001-02-02

    Expression of high quantities of alfalfa hydroperoxide lyase in Escherichia coli made it possible to study its active site and structure in more detail. Circular dichroism (CD) spectra showed that hydroperoxide lyase consists for about 75% of alpha-helices. Electron paramagnetic resonance (EPR) spectra confirmed its classification as a cytochrome P450 enzyme. The positive influence of detergents on the enzyme activity is paralleled by a spin state transition of the heme Fe(III) from low to high spin. EPR and CD spectra showed that detergents induce a subtle conformational change, which might result in improved substrate binding. Because hydroperoxide lyase is thought to be a membrane bound protein and detergents mimic a membrane environment, the more active, high spin form likely represents the in vivo conformation. Furthermore, the spin state appeared to be temperature-dependent, with the low spin state favored at low temperature. Point mutants of the highly conserved cysteine in domain D indicated that this residue might be involved in heme binding.

  15. Anisotropy Changes of a Fluorescent Probe during the Micellar Growth and Clouding of a Nonionic Detergent.

    PubMed

    Komaromy-Hiller; von Wandruszka R

    1996-01-15

    The effects of temperature and Triton X-114 (TX-114) concentration on the fluorescence anisotropy of perylene were investigated before and after detergent clouding. The measured anisotropy values were used to estimate the microviscosity of the micellar interior. In the lower detergent concentration range, an anisotropy maximum was observed at the critical micelle concentration (CMC), while the values decreased in the range immediately above the CMC. This was ascribed to the micellar volume increase, which, in the case of TX-114, was not accompanied by a more ordered internal environment. A gradual decrease of anisotropy and microviscosity with increasing temperature below the cloud point was observed. At the cloud point, no abrupt changes were found to occur. Compared to detergents with more flexible hydrophobic moieties, TX-114 micelles have a relatively ordered micellar interior indicated by the microviscosity and calculated fusion energy values. In the separated micellar phase formed after clouding, the probe anisotropy increased as water was eliminated at higher temperatures.

  16. Fast and accurate determination of the detergent efficiency by optical fiber sensors

    NASA Astrophysics Data System (ADS)

    Patitsa, Maria; Pfeiffer, Helge; Wevers, Martine

    2011-06-01

    An optical fiber sensor was developed to control the cleaning efficiency of surfactants. Prior to the measurements, the sensing part of the probe is covered with a uniform standardized soil layer (lipid multilayer), and a gold mirror is deposited at the end of the optical fiber. For the lipid multilayer deposition on the fiber, Langmuir-Blodgett technique was used and the progress of deposition was followed online by ultraviolet spectroscopy. The invention provides a miniaturized Surface Plasmon Resonance dip-sensor for automated on-line testing that can replace the cost and time consuming existing methods and develop a breakthrough in detergent testing in combining optical sensing, surface chemistry and automated data acquisition. The sensor is to be used to evaluate detergency of different cleaning products and also indicate how formulation, concentration, lipid nature and temperature affect the cleaning behavior of a surfactant.

  17. Using detergent to enhance detection sensitivity of African trypanosomes in human CSF and blood by loop-mediated isothermal amplification (LAMP).

    PubMed

    Grab, Dennis J; Nikolskaia, Olga V; Inoue, Noboru; Thekisoe, Oriel M M; Morrison, Liam J; Gibson, Wendy; Dumler, J Stephen

    2011-08-01

    The loop-mediated isothermal amplification (LAMP) assay, with its advantages of simplicity, rapidity and cost effectiveness, has evolved as one of the most sensitive and specific methods for the detection of a broad range of pathogenic microorganisms including African trypanosomes. While many LAMP-based assays are sufficiently sensitive to detect DNA well below the amount present in a single parasite, the detection limit of the assay is restricted by the number of parasites present in the volume of sample assayed; i.e. 1 per µL or 10(3) per mL. We hypothesized that clinical sensitivities that mimic analytical limits based on parasite DNA could be approached or even obtained by simply adding detergent to the samples prior to LAMP assay. For proof of principle we used two different LAMP assays capable of detecting 0.1 fg genomic DNA (0.001 parasite). The assay was tested on dilution series of intact bloodstream form Trypanosoma brucei rhodesiense in human cerebrospinal fluid (CSF) or blood with or without the addition of the detergent Triton X-100 and 60 min incubation at ambient temperature. With human CSF and in the absence of detergent, the LAMP detection limit for live intact parasites using 1 µL of CSF as the source of template was at best 10(3) parasites/mL. Remarkably, detergent enhanced LAMP assay reaches sensitivity about 100 to 1000-fold lower; i.e. 10 to 1 parasite/mL. Similar detergent-mediated increases in LAMP assay analytical sensitivity were also found using DNA extracted from filter paper cards containing blood pretreated with detergent before card spotting or blood samples spotted on detergent pretreated cards. This simple procedure for the enhanced detection of live African trypanosomes in biological fluids by LAMP paves the way for the adaptation of LAMP for the economical and sensitive diagnosis of other protozoan parasites and microorganisms that cause diseases that plague the developing world.

  18. Effect of some detergents, humate, and composition of seedbed on crop of tomato plants in a hydroponic culture

    NASA Technical Reports Server (NTRS)

    Guminka, A. Z.; Gracz-Nalepka, M.; Lukasiewicz, B.; Sobolewicz, E.; Turkiewicz, I. T.

    1978-01-01

    It is established that single detergent doses distinctly stimulate vegetative development of plants in the initial stage when humates are available. When detergents are applied every four weeks in a hydroponic culture, in which the seedbed does not contain active humates, the crop is reduced by 50%. This adverse effect does not occur when the seedbed is a mixture of brown coal and peat.

  19. Zwitterionic polymer functionalization of polysulfone membrane with improved antifouling property and blood compatibility by combination of ATRP and click chemistry.

    PubMed

    Xiang, Tao; Lu, Ting; Xie, Yi; Zhao, Wei-Feng; Sun, Shu-Dong; Zhao, Chang-Sheng

    2016-08-01

    The chemical compositions are very important for designing blood-contacting membranes with good antifouling property and blood compatibility. In this study, we propose a method combining ATRP and click chemistry to introduce zwitterionic polymer of poly(sulfobetaine methacrylate) (PSBMA), negatively charged polymers of poly(sodium methacrylate) (PNaMAA) and/or poly(sodium p-styrene sulfonate) (PNaSS), to improve the antifouling property and blood compatibility of polysulfone (PSf) membranes. Attenuated total reflectance-Fourier transform infrared spectra, X-ray photoelectron spectroscopy and water contact angle results confirmed the successful grafting of the functional polymers. The antifouling property and blood compatibility of the modified membranes were systematically investigated. The zwitterionic polymer (PSBMA) grafted membranes showed good resistance to protein adsorption and bacterial adhesion; the negatively charged polymer (PNaSS or PNaMAA) grafted membranes showed improved blood compatibility, especially the anticoagulant property. Moreover, the PSBMA/PNaMAA modified membrane showed both antifouling property and anticoagulant property, and exhibited a synergistic effect in inhibiting blood coagulation. The functionalization of membrane surfaces by a combination of ATRP and click chemistry is demonstrated as an effective route to improve the antifouling property and blood compatibility of membranes in blood-contact. Copyright © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  20. Investigation of the retention/pH profile of zwitterionic fluoroquinolones in reversed-phase and ion-interaction high performance liquid chromatography.

    PubMed

    Pistos, C; Tsantili-Kakoulidou, A; Koupparis, M

    2005-09-15

    The retention/pH profiles of three fluoroquinolones, ofloxacin, norfloxacin and ciprofloxacin, was investigated by means of reversed-phase high performance liquid chromatography (RP-HPLC) and reversed-phase ion-interaction chromatography (RP-IIC), using an octadecylsilane stationary phase and acetonitrile as organic modifier. Sodium hexanesulphonate and tetrabutylammonium hydroxide were used as sources of counter ions in ion-interaction chromatography. The retention/pH profiles under in RP-HPLC were compared to the corresponding lipophilicity/pH profiles. Despite the rather hydrophilic nature of the three fluoroquinolones positive retention factors were obtained while there was a shift of the retention maximum towards more acidic pH values. This behavior was attributed mainly to non-hydrophobic silanophilic interactions with the silanized silica gel material of the stationary phase. In ion-interaction chromatography the effect of counter ions over a broad pH range was found to be ruled rather by the ion pair formation in the mobile phase which led to a drastic decrease in retention as a consequence of the disruption of the zwitterionic structure and thereupon the deliberation of a net charge in the molecules. At pH values at which zwitterionic structure was not favored both the ion-exchange and ion pair formation mechanisms were assumed to contribute to the retention.