Fixed-Charge Atomistic Force Fields for Molecular Dynamics Simulations in the Condensed Phase: An Overview.

Science.gov (United States)

Riniker, Sereina

2018-03-26

In molecular dynamics or Monte Carlo simulations, the interactions between the particles (atoms) in the system are described by a so-called force field. The empirical functional form of classical fixed-charge force fields dates back to 1969 and remains essentially unchanged. In a fixed-charge force field, the polarization is not modeled explicitly, i.e. the effective partial charges do not change depending on conformation and environment. This simplification allows, however, a dramatic reduction in computational cost compared to polarizable force fields and in particular quantum-chemical modeling. The past decades have shown that simulations employing carefully parametrized fixed-charge force fields can provide useful insights into biological and chemical questions. This overview focuses on the four major force-field families, i.e. AMBER, CHARMM, GROMOS, and OPLS, which are based on the same classical functional form and are continuously improved to the present day. The overview is aimed at readers entering the field of (bio)molecular simulations. More experienced users may find the comparison and historical development of the force-field families interesting.

Macroporous modified poly (vinyl alcohol) hydrogels with charged groups for tissue engineering: Preparation and in vitro evaluation

Energy Technology Data Exchange (ETDEWEB)

Drozdova, Maria G., E-mail: drozdovamg@gmail.com [Polymers for Biology Laboratory, Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of Russian Academy of Sciences, Miklukho-Maklaya str., 16/10, Moscow 117997 (Russian Federation); Zaytseva-Zotova, Daria S. [Polymers for Biology Laboratory, Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of Russian Academy of Sciences, Miklukho-Maklaya str., 16/10, Moscow 117997 (Russian Federation); Akasov, Roman A. [Polymers for Biology Laboratory, Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of Russian Academy of Sciences, Miklukho-Maklaya str., 16/10, Moscow 117997 (Russian Federation); Sechenov First Moscow State Medical University, Institute for Regenerative Medicine, Trubetskaya str., 8/2, Moscow 119048 (Russian Federation); Golunova, Anna S.; Artyukhov, Alexander A. [D. Mendeleyev University of Chemical Technology of Russia, Miusskaya Square 9, Moscow 125047 (Russian Federation); Udartseva, Olga O.; Andreeva, Elena R. [Institute of Biomedical Problems of Russian Academy of Sciences, Khoroshevskoe Shosse 76a, Moscow 123007 (Russian Federation); Lisovyy, Denis E.; Shtilman, Michael I. [D. Mendeleyev University of Chemical Technology of Russia, Miusskaya Square 9, Moscow 125047 (Russian Federation); Markvicheva, Elena A. [Polymers for Biology Laboratory, Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry of Russian Academy of Sciences, Miklukho-Maklaya str., 16/10, Moscow 117997 (Russian Federation)

2017-06-01

Poly(vinyl alcohol) (PVA) hydrogels are widely employed for various biomedical applications, including tissue engineering, due to their biocompatibility, high water solubility, low protein adsorption, and chemical stability. However, non-charged surface of PVA-based hydrogels is not optimal for cell adhesion and spreading. Here, cross-linked macroporous hydrogels based on low molecular weight acrylated PVA (Acr-PVA) was synthesized by modification of the pendant alcohol groups on the PVA with glycidyl methacrylate (GMA). To enhance cell affinity, charged groups were introduced to the hydrogel composition. For this purpose, Acr-PVA was copolymerized with either negatively charged acrylic acid (AA) or positively charged 2-(diethylamino) ethyl methacrylate (DEAEMA) monomers. A surface charge of the obtained hydrogels was found to be in function of the co-monomer type and content. Confocal microscopy observations confirmed that adhesion and spreading of both mouse fibroblasts (L929) and human mesenchymal stem cells (hMSC) on the modified Acr-PVA-AA and Acr-PVA-DEAEMA hydrogels were better than those on the non-modified Acr-PVA hydrogel. The increase of DEAEMA monomer content from 5 to 15 mol% resulted in the enhancement of cell viability which was 1.5-fold higher for Acr-PVA-DEAEMA-15 hydrogel than that of the non-modified Acr-PVA hydrogel sample. - Highlights: • To enhance cell affinity, acrylated PVA hydrogel was modified with AA or DEAEMA monomers. • Cell adhesion and spreading were found to depend on the co-monomer type and content. • Proliferation of L929 fibroblasts and stem cells increased on the modified hydrogels.

Macroporous modified poly (vinyl alcohol) hydrogels with charged groups for tissue engineering: Preparation and in vitro evaluation

International Nuclear Information System (INIS)

Drozdova, Maria G.; Zaytseva-Zotova, Daria S.; Akasov, Roman A.; Golunova, Anna S.; Artyukhov, Alexander A.; Udartseva, Olga O.; Andreeva, Elena R.; Lisovyy, Denis E.; Shtilman, Michael I.; Markvicheva, Elena A.

2017-01-01

Poly(vinyl alcohol) (PVA) hydrogels are widely employed for various biomedical applications, including tissue engineering, due to their biocompatibility, high water solubility, low protein adsorption, and chemical stability. However, non-charged surface of PVA-based hydrogels is not optimal for cell adhesion and spreading. Here, cross-linked macroporous hydrogels based on low molecular weight acrylated PVA (Acr-PVA) was synthesized by modification of the pendant alcohol groups on the PVA with glycidyl methacrylate (GMA). To enhance cell affinity, charged groups were introduced to the hydrogel composition. For this purpose, Acr-PVA was copolymerized with either negatively charged acrylic acid (AA) or positively charged 2-(diethylamino) ethyl methacrylate (DEAEMA) monomers. A surface charge of the obtained hydrogels was found to be in function of the co-monomer type and content. Confocal microscopy observations confirmed that adhesion and spreading of both mouse fibroblasts (L929) and human mesenchymal stem cells (hMSC) on the modified Acr-PVA-AA and Acr-PVA-DEAEMA hydrogels were better than those on the non-modified Acr-PVA hydrogel. The increase of DEAEMA monomer content from 5 to 15 mol% resulted in the enhancement of cell viability which was 1.5-fold higher for Acr-PVA-DEAEMA-15 hydrogel than that of the non-modified Acr-PVA hydrogel sample. - Highlights: • To enhance cell affinity, acrylated PVA hydrogel was modified with AA or DEAEMA monomers. • Cell adhesion and spreading were found to depend on the co-monomer type and content. • Proliferation of L929 fibroblasts and stem cells increased on the modified hydrogels.

Surface charge effects in protein adsorption on nanodiamonds

Science.gov (United States)

Aramesh, M.; Shimoni, O.; Ostrikov, K.; Prawer, S.; Cervenka, J.

2015-03-01

Understanding the interaction of proteins with charged diamond nanoparticles is of fundamental importance for diverse biomedical applications. Here we present a thorough study of protein binding, adsorption kinetics and structure on strongly positively (hydrogen-terminated) and negatively (oxygen-terminated) charged nanodiamond particles using a quartz crystal microbalance by dissipation and infrared spectroscopy. By using two model proteins (bovine serum albumin and lysozyme) of different properties (charge, molecular weight and rigidity), the main driving mechanism responsible for the protein binding to the charged nanoparticles was identified. Electrostatic interactions were found to dominate the protein adsorption dynamics, attachment and conformation. We developed a simple electrostatic model that can qualitatively explain the observed adsorption behaviour based on charge-induced pH modifications near the charged nanoparticle surfaces. Under neutral conditions, the local pH around the positively and negatively charged nanodiamonds becomes very high (11-12) and low (1-3) respectively, which has a profound impact on the protein charge, hydration and affinity to the nanodiamonds. Small proteins (lysozyme) were found to form multilayers with significant conformational changes to screen the surface charge, while larger proteins (albumin) formed monolayers with minor conformational changes. The findings of this study provide a step forward toward understanding and eventually predicting nanoparticle interactions with biofluids.Understanding the interaction of proteins with charged diamond nanoparticles is of fundamental importance for diverse biomedical applications. Here we present a thorough study of protein binding, adsorption kinetics and structure on strongly positively (hydrogen-terminated) and negatively (oxygen-terminated) charged nanodiamond particles using a quartz crystal microbalance by dissipation and infrared spectroscopy. By using two model proteins

Surface charge effects in protein adsorption on nanodiamonds.

Science.gov (United States)

Aramesh, M; Shimoni, O; Ostrikov, K; Prawer, S; Cervenka, J

2015-03-19

Understanding the interaction of proteins with charged diamond nanoparticles is of fundamental importance for diverse biomedical applications. Here we present a thorough study of protein binding, adsorption kinetics and structure on strongly positively (hydrogen-terminated) and negatively (oxygen-terminated) charged nanodiamond particles using a quartz crystal microbalance by dissipation and infrared spectroscopy. By using two model proteins (bovine serum albumin and lysozyme) of different properties (charge, molecular weight and rigidity), the main driving mechanism responsible for the protein binding to the charged nanoparticles was identified. Electrostatic interactions were found to dominate the protein adsorption dynamics, attachment and conformation. We developed a simple electrostatic model that can qualitatively explain the observed adsorption behaviour based on charge-induced pH modifications near the charged nanoparticle surfaces. Under neutral conditions, the local pH around the positively and negatively charged nanodiamonds becomes very high (11-12) and low (1-3) respectively, which has a profound impact on the protein charge, hydration and affinity to the nanodiamonds. Small proteins (lysozyme) were found to form multilayers with significant conformational changes to screen the surface charge, while larger proteins (albumin) formed monolayers with minor conformational changes. The findings of this study provide a step forward toward understanding and eventually predicting nanoparticle interactions with biofluids.

Protein Charge and Mass Contribute to the Spatio-temporal Dynamics of Protein-Protein Interactions in a Minimal Proteome

Science.gov (United States)

Xu, Yu; Wang, Hong; Nussinov, Ruth; Ma, Buyong

2013-01-01

We constructed and simulated a ‘minimal proteome’ model using Langevin dynamics. It contains 206 essential protein types which were compiled from the literature. For comparison, we generated six proteomes with randomized concentrations. We found that the net charges and molecular weights of the proteins in the minimal genome are not random. The net charge of a protein decreases linearly with molecular weight, with small proteins being mostly positively charged and large proteins negatively charged. The protein copy numbers in the minimal genome have the tendency to maximize the number of protein-protein interactions in the network. Negatively charged proteins which tend to have larger sizes can provide large collision cross-section allowing them to interact with other proteins; on the other hand, the smaller positively charged proteins could have higher diffusion speed and are more likely to collide with other proteins. Proteomes with random charge/mass populations form less stable clusters than those with experimental protein copy numbers. Our study suggests that ‘proper’ populations of negatively and positively charged proteins are important for maintaining a protein-protein interaction network in a proteome. It is interesting to note that the minimal genome model based on the charge and mass of E. Coli may have a larger protein-protein interaction network than that based on the lower organism M. pneumoniae. PMID:23420643

13 CFR 120.213 - What fixed interest rates may a Lender charge?

Science.gov (United States)

2010-01-01

... Lender charge? 120.213 Section 120.213 Business Credit and Assistance SMALL BUSINESS ADMINISTRATION... have a reasonable fixed interest rate. SBA periodically publishes the maximum allowable rate in the... government determines the interest rate on direct loans. SBA publishes the rate periodically in the Federal...

Iterated local search and record-to-record travel applied to the fixed charge transportation problem

DEFF Research Database (Denmark)

Andersen, Jeanne; Klose, Andreas

The fixed charge transportation problem (FCTP) is a well-known and difficult optimization problem with lots of applications in logistics. It consists in finding a minimum cost network flow from a set of suppliers to a set of customers. Beside costs proportional to quantities transported......, transportation costs do, however, include a fixed charge. Iterated local search and record-to-record travel are both simple local search based meta-heuristics that, to our knowledge, not yet have been applied to the FCTP. In this paper, we apply both types of search strategies and combine them into a single...

Identification of multiply charged proteins and amino acid clusters by liquid nitrogen assisted spray ionization mass spectrometry.

Science.gov (United States)

Kumar Kailasa, Suresh; Hasan, Nazim; Wu, Hui-Fen

2012-08-15

The development of liquid nitrogen assisted spray ionization mass spectrometry (LNASI MS) for the analysis of multiply charged proteins (insulin, ubiquitin, cytochrome c, α-lactalbumin, myoglobin and BSA), peptides (glutathione, HW6, angiotensin-II and valinomycin) and amino acid (arginine) clusters is described. The charged droplets are formed by liquid nitrogen assisted sample spray through a stainless steel nebulizer and transported into mass analyzer for the identification of multiply charged protein ions. The effects of acids and modifier volumes for the efficient ionization of the above analytes in LNASI MS were carefully investigated. Multiply charged proteins and amino acid clusters were effectively identified by LNASI MS. The present approach can effectively detect the multiply charged states of cytochrome c at 400 nM. A comparison between LNASI and ESI, CSI, SSI and V-EASI methods on instrumental conditions, applied temperature and observed charge states for the multiply charged proteins, shows that the LNASI method produces the good quality spectra of amino acid clusters at ambient conditions without applied any electric field and heat. To date, we believe that the LNASI method is the most simple, low cost and provided an alternative paradigm for production of multiply charged ions by LNASI MS, just as ESI-like ions yet no need for applying any electrical field and it could be operated at low temperature for generation of highly charged protein/peptide ions. Copyright © 2012 Elsevier B.V. All rights reserved.

Nanomechanics of layer-by-layer polyelectrolyte complexes: a manifestation of ionic cross-links and fixed charges.

Science.gov (United States)

Han, Biao; Chery, Daphney R; Yin, Jie; Lu, X Lucas; Lee, Daeyeon; Han, Lin

2016-01-28

This study investigates the roles of two distinct features of ionically cross-linked polyelectrolyte networks - ionic cross-links and fixed charges - in determining their nanomechanical properties. The layer-by-layer assembled poly(allylamine hydrochloride)/poly(acrylic acid) (PAH/PAA) network is used as the model material. The densities of ionic cross-links and fixed charges are modulated through solution pH and ionic strength (IS), and the swelling ratio, elastic and viscoelastic properties are quantified via an array of atomic force microscopy (AFM)-based nanomechanical tools. The roles of ionic cross-links are underscored by the distinctive elastic and viscoelastic nanomechanical characters observed here. First, as ionic cross-links are highly sensitive to solution conditions, the instantaneous modulus, E0, exhibits orders-of-magnitude changes upon pH- and IS-governed swelling, distinctive from the rubber elasticity prediction based on permanent covalent cross-links. Second, ionic cross-links can break and self-re-form, and this mechanism dominates force relaxation of PAH/PAA under a constant indentation depth. In most states, the degree of relaxation is >90%, independent of ionic cross-link density. The importance of fixed charges is highlighted by the unexpectedly more elastic nature of the network despite low ionic cross-link density at pH 2.0, IS 0.01 M. Here, the complex is a net charged, loosely cross-linked, where the degree of relaxation is attenuated to ≈50% due to increased elastic contribution arising from fixed charge-induced Donnan osmotic pressure. In addition, this study develops a new method for quantifying the thickness of highly swollen polymer hydrogel films. It also underscores important technical considerations when performing nanomechanical tests on highly rate-dependent polymer hydrogel networks. These results provide new insights into the nanomechanical characters of ionic polyelectrolyte complexes, and lay the ground for further

Alternate mutation based artificial immune algorithm for step fixed charge transportation problem

Directory of Open Access Journals (Sweden)

Mahmoud Moustafa El-Sherbiny

2012-07-01

Full Text Available Step fixed charge transportation problem (SFCTP is considered as a special version of the fixed-charge transportation problem (FCTP. In SFCTP, the fixed cost is incurred for every route that is used in the solution and is proportional to the amount shipped. This cost structure causes the value of the objective function to behave like a step function. Both FCTP and SFCTP are considered to be NP-hard problems. While a lot of research has been carried out concerning FCTP, not much has been done concerning SFCTP. This paper introduces an alternate Mutation based Artificial Immune (MAI algorithm for solving SFCTPs. The proposed MAI algorithm solves both balanced and unbalanced SFCTP without introducing a dummy supplier or a dummy customer. In MAI algorithm a coding schema is designed and procedures are developed for decoding such schema and shipping units. MAI algorithm guarantees the feasibility of all the generated solutions. Due to the significant role of mutation function on the MAI algorithm’s quality, 16 mutation functions are presented and their performances are compared to select the best one. For this purpose, forty problems with different sizes have been generated at random and then a robust calibration is applied using the relative percentage deviation (RPD method. Through two illustrative problems of different sizes the performance of the MAI algorithm has been compared with most recent methods.

Modified intuitionistic fuzzy metric spaces and some fixed point theorems

International Nuclear Information System (INIS)

Saadati, R.; Sedghi, S.; Shobe, N.

2008-01-01

Since the intuitionistic fuzzy metric space has extra conditions (see [Gregori V, Romaguera S, Veereamani P. A note on intuitionistic fuzzy metric spaces. Chaos, Solitons and Fractals 2006;28:902-5]). In this paper, we consider modified intuitionistic fuzzy metric spaces and prove some fixed point theorems in these spaces. All the results presented in this paper are new

A Soluble, Folded Protein without Charged Amino Acid Residues

DEFF Research Database (Denmark)

Højgaard, Casper; Kofoed, Christian; Espersen, Roall

2016-01-01

side chains can maintain solubility, stability, and function. As a model, we used a cellulose-binding domain from Cellulomonas fimi, which, among proteins of more than 100 amino acids, presently is the least charged in the Protein Data Bank, with a total of only four titratable residues. We find......Charges are considered an integral part of protein structure and function, enhancing solubility and providing specificity in molecular interactions. We wished to investigate whether charged amino acids are indeed required for protein biogenesis and whether a protein completely free of titratable...... that the protein shows a surprising resilience toward extremes of pH, demonstrating stability and function (cellulose binding) in the pH range from 2 to 11. To ask whether the four charged residues present were required for these properties of this protein, we altered them to nontitratable ones. Remarkably...

A Soluble, Folded Protein without Charged Amino Acid Residues

DEFF Research Database (Denmark)

Højgaard, Casper; Kofoed, Christian; Espersen, Roall

2016-01-01

Charges are considered an integral part of protein structure and function, enhancing solubility and providing specificity in molecular interactions. We wished to investigate whether charged amino acids are indeed required for protein biogenesis and whether a protein completely free of titratable...... side chains can maintain solubility, stability, and function. As a model, we used a cellulose-binding domain from Cellulomonas fimi, which, among proteins of more than 100 amino acids, presently is the least charged in the Protein Data Bank, with a total of only four titratable residues. We find...

Chromatin-modifying proteins in cancer

DEFF Research Database (Denmark)

Fog, Cathrine K; Jensen, Klaus T; Lund, Anders Henrik

2007-01-01

-despite the fact that all cells in the organism contain the same genetic information. A large amount of data gathered over the last decades has demonstrated that deregulation of chromatin-modifying proteins is etiologically involved in the development and progression of cancer. Here we discuss how epigenetic...... alterations influence cancer development and review known cancer-associated alterations in chromatin-modifying proteins....

Using modified soy protein to enhance foaming of egg white protein.

Science.gov (United States)

Wang, Guang; Troendle, Molly; Reitmeier, Cheryll A; Wang, Tong

2012-08-15

It is well known that the foaming properties of egg white protein are significantly reduced when a small amount of yolk is mixed in the white. To improve foaming properties of yolk-contaminated egg white protein, soy protein isolate (SPI) and egg proteins were modified to make basic proteins, and effects of these modified proteins on egg white foaming were evaluated in a model and an angel cake system. SPI and egg yolk proteins were modified to have an isoelectric point of 10, and sonication was used to increase protein dispersibility after the ethyl esterification reaction. However, only the addition of sonicated and modified SPI (SMSPI) showed improvement of foaming in the 5% egg protein model system with 0.4% yolk addition. SMSPI was then used in making angel food cake to examine whether the cake performance reduction due to yolk contamination of the white would be restored by such alkaline protein. Cake performance was improved when cream of tartar was used together with SMSPI. Basic soy protein can be made and used to improve egg white foaming properties and cake performance. Copyright © 2012 Society of Chemical Industry.

Hydration shells exchange charge with their protein

DEFF Research Database (Denmark)

Abitan, Haim; Lindgård, Per-Anker; Nielsen, Bjørn Gilbert

2010-01-01

. In our experiments, the amplitude of an ultrasonic pressure wave is gradually increased (0–20 atm) while we simultaneously measure the Raman spectra from the hydrated protein (β-lactoglobulin and lysozyme). We detected two types of spectral changes: first, up to 70% increase in the intensity......Investigation of the interaction between a protein and its hydration shells is an experimental and theoretical challenge. Here, we used ultrasonic pressure waves in aqueous solutions of a protein to explore the conformational states of the protein and its interaction with its hydration shells...... the presence of an ultrasonic pressure, a protein and its hydration shells are in thermodynamic and charge equilibrium, i.e. a protein and its hydration shells exchange charges. The ultrasonic wave disrupts these equilibria which are regained within 30–45 min after the ultrasonic pressure is shut off....

Proteins mediating intra- and intercellular transport of lipids and lipid-modified proteins

NARCIS (Netherlands)

Neumann, S.

2008-01-01

Proteins mediating intra- and intercellular transport of lipids and lipid-modified proteins In this thesis, I studied the intra- and intercellular transport of lipidic molecules, in particular glycosphingolipids and lipid-modified proteins. The first part focuses on the intracellular transport of

Do plasma proteins distinguish between liposomes of varying charge density?

KAUST Repository

Capriotti, Anna Laura

2012-03-01

Cationic liposomes (CLs) are one of the most employed nonviral nanovector systems in gene therapy. However, their transfection efficiency is strongly affected by interactions with plasma components, that lead to the formation of a "protein corona" onto CL surface. The interactions between nanoparticles entering the body and biomolecules have an essential role for their biodistribution. Because the knowledge of proteins adsorbed onto vector surface could be useful in the screening of new, more efficient and more biocompatible liposomal formulations, the behavior of three CLs with different membrane charge densities was investigated. The proteins of the three coronas were identified by nano-liquid chromatography-tandem mass spectrometry, and quantified with label-free spectral counting strategy. Fibrinogen displayed higher association with CLs with high membrane charge density, while apolipoproteins and C4b-binding protein with CLs with low membrane charge density. These results are discussed in terms of the different lipid compositions of CLs and may have a deep biological impact for in vivo applications. Surface charge of nanoparticles is emerging as a relevant factor determining the corona composition after interaction with plasma proteins. Remarkably, it is also shown that the charge of the protein corona formed around CLs is strongly related to their membrane charge density. © 2012 Elsevier B.V.

Huntingtin interacting proteins are genetic modifiers of neurodegeneration.

Directory of Open Access Journals (Sweden)

Linda S Kaltenbach

2007-05-01

Full Text Available Huntington's disease (HD is a fatal neurodegenerative condition caused by expansion of the polyglutamine tract in the huntingtin (Htt protein. Neuronal toxicity in HD is thought to be, at least in part, a consequence of protein interactions involving mutant Htt. We therefore hypothesized that genetic modifiers of HD neurodegeneration should be enriched among Htt protein interactors. To test this idea, we identified a comprehensive set of Htt interactors using two complementary approaches: high-throughput yeast two-hybrid screening and affinity pull down followed by mass spectrometry. This effort led to the identification of 234 high-confidence Htt-associated proteins, 104 of which were found with the yeast method and 130 with the pull downs. We then tested an arbitrary set of 60 genes encoding interacting proteins for their ability to behave as genetic modifiers of neurodegeneration in a Drosophila model of HD. This high-content validation assay showed that 27 of 60 orthologs tested were high-confidence genetic modifiers, as modification was observed with more than one allele. The 45% hit rate for genetic modifiers seen among the interactors is an order of magnitude higher than the 1%-4% typically observed in unbiased genetic screens. Genetic modifiers were similarly represented among proteins discovered using yeast two-hybrid and pull-down/mass spectrometry methods, supporting the notion that these complementary technologies are equally useful in identifying biologically relevant proteins. Interacting proteins confirmed as modifiers of the neurodegeneration phenotype represent a diverse array of biological functions, including synaptic transmission, cytoskeletal organization, signal transduction, and transcription. Among the modifiers were 17 loss-of-function suppressors of neurodegeneration, which can be considered potential targets for therapeutic intervention. Finally, we show that seven interacting proteins from among 11 tested were able to

[Interaction of protein with charged colloidal particles].

Science.gov (United States)

Durdenko, E V; Kuznetsova, S M; Basova, L V; Tikhonenko, S A; Saburova, E A

2011-01-01

The functional state of three proteins of different molecular weight (urease, lactate dehydrogenase, and hemoglobin) in the presence of the linear polyelectrolytes poly(allylamine hydrochloride) (PAA) and sodium poly(styrenesulfonate) (PSS) in the dissolved state and of the same polyelectrolytes bound to the surface of microspheres has been investigated. Microspheres were prepared by consecutive absorption of oppositely charged polyelectrolytes so that the outer layer of the shell was PAA for the acidic protein urease, and PSS for the alkaline proteins LDH and hemoglobin. It was shown that the dissolved polyelectrolyte completely inactivates all three proteins within one minute with a slight difference in the time constant. (By Hb inactivation are conventionally meant changes in the heme environment observed from the spectrum in the Soret band.) In the presence of microspheres, the proteins were adsorbed on their surface; in this case, more than 95% of the activity was retained within two hours. The proportion of the protein adsorbed on microspheres accounted for about 98% for urease, 72% for Hb, and 35% for LDH, as determined from the tryptophan fluorescence data. The interaction of hemoglobin with another type of charged colloidal particles, phospholipid vesicles, leads to the destruction of the tertiary structure of the protein, which made itself evident in the optical absorption spectra in the Soret band, as well as the spectra of tryptophan fluorescence and circular dichroism. In this case, according to circular dichroism, the percentage of alpha-helical structure of Hb was maintained. The differences in the physical and chemical mechanisms of interaction of proteins with these two types of charged colloidal particles that leads to differences in the degree of denaturing effects are discussed.

Insulator-semiconductor interface fixed charges in AlGaN/GaN metal-insulator-semiconductor devices with Al2O3 or AlTiO gate dielectrics

Science.gov (United States)

Le, Son Phuong; Nguyen, Duong Dai; Suzuki, Toshi-kazu

2018-01-01

We have investigated insulator-semiconductor interface fixed charges in AlGaN/GaN metal-insulator-semiconductor (MIS) devices with Al2O3 or AlTiO (an alloy of Al2O3 and TiO2) gate dielectrics obtained by atomic layer deposition on AlGaN. Analyzing insulator-thickness dependences of threshold voltages for the MIS devices, we evaluated positive interface fixed charges, whose density at the AlTiO/AlGaN interface is significantly lower than that at the Al2O3/AlGaN interface. This and a higher dielectric constant of AlTiO lead to rather shallower threshold voltages for the AlTiO gate dielectric than for Al2O3. The lower interface fixed charge density also leads to the fact that the two-dimensional electron concentration is a decreasing function of the insulator thickness for AlTiO, whereas being an increasing function for Al2O3. Moreover, we discuss the relationship between the interface fixed charges and interface states. From the conductance method, it is shown that the interface state densities are very similar at the Al2O3/AlGaN and AlTiO/AlGaN interfaces. Therefore, we consider that the lower AlTiO/AlGaN interface fixed charge density is not owing to electrons trapped at deep interface states compensating the positive fixed charges and can be attributed to a lower density of oxygen-related interface donors.

Photochemistry of modified proteins benzophenone-containing bovine serum albumin

International Nuclear Information System (INIS)

Mariano, P.S.; Glover, G.I.; Wilkinson, T.J.

1976-01-01

The results of exploratory and mechanistic studies of the photochemistry of poly-p-benzoyl-acetimido-bovine serum albumin, a modified protein containing photoreactive and photosensitizing groups, are reported. Specifically described are recent findings concerning (1) the synthesis and characterization of a modified bovine serum albumin that contains benzophenone-like moieties, (2) the photochemistry of this modified protein which appeared to involve photoreductive coupling of the benzophenone chromophores to the protein backbone, and (3) triplet energy transfer from modified bovine serum albumin to small molecule acceptors resulting in quenching of the photoreaction. (author)

Advanced glycation end product (AGE) modified proteins in tears of diabetic patients.

Science.gov (United States)

Zhao, Zhenjun; Liu, Jingfang; Shi, Bingyin; He, Shuixiang; Yao, Xiaoli; Willcox, Mark D P

2010-08-11

High glucose level in diabetic patients may lead to advanced glycation end product (AGE) modified proteins. This study investigated AGE modified proteins in tears and compared their levels in diabetic patients (DM) with non-diabetic controls (CTL). Basal tears were collected from DM with (DR) or without (DNR) retinopathy and CTL. Total AGE modified proteins were detected quantitatively by a dot immunobinding assay. The AGE modified proteins were separated in 1D- and 2D-SDS gels and detected by western-blotting. The individual AGE modified proteins were also compared between groups using densitometry. Compared with the CTL group, tear concentrations of AGE modified proteins were significantly elevated in DR and DNR groups. The concentration of AGE modified proteins in diabetic tears were positively correlated with AGE modified hemoglobin (HbA1c) and postprandial blood glucose level (PBG). Western blotting of AGE modified proteins from 1D-SDS gels showed several bands, the major one at around 60 kDa. The intensities of AGE modified protein bands were higher in DM tears than in CTL tears. Western blotting from 2D-SDS gels showed a strongly stained horizontal strip, which corresponded to the major band in 1D-SDS gels. Most of the other AGE modified protein species were within molecular weight of 30-60 kDa, PI 5.2-7.0. Densitometry analysis demonstrated several AGE modified proteins were elevated in DR or DNR tears. Total and some individual AGE modified proteins were elevated in DM tears. AGE modified proteins in tears may be used as biomarkers to diagnose diabetes and/or diabetic retinopathy.

Adhesives from modified soy protein

Science.gov (United States)

Sun, Susan [Manhattan, KS; Wang, Donghai [Manhattan, KS; Zhong, Zhikai [Manhattan, KS; Yang, Guang [Shanghai, CN

2008-08-26

The present invention provides useful adhesive compositions having similar adhesive properties to conventional UF and PPF resins. The compositions generally include a protein portion and modifying ingredient portion selected from the group consisting of carboxyl-containing compounds, aldehyde-containing compounds, epoxy group-containing compounds, and mixtures thereof. The composition is preferably prepared at a pH level at or near the isoelectric point of the protein. In other preferred forms, the adhesive composition includes a protein portion and a carboxyl-containing group portion.

Fast Atomic Charge Calculation for Implementation into a Polarizable Force Field and Application to an Ion Channel Protein

Directory of Open Access Journals (Sweden)

Raiker Witter

2015-01-01

Full Text Available Polarization of atoms plays a substantial role in molecular interactions. Class I and II force fields mostly calculate with fixed atomic charges which can cause inadequate descriptions for highly charged molecules, for example, ion channels or metalloproteins. Changes in charge distributions can be included into molecular mechanics calculations by various methods. Here, we present a very fast computational quantum mechanical method, the Bond Polarization Theory (BPT. Atomic charges are obtained via a charge calculation method that depend on the 3D structure of the system in a similar way as atomic charges of ab initio calculations. Different methods of population analysis and charge calculation methods and their dependence on the basis set were investigated. A refined parameterization yielded excellent correlation of R=0.9967. The method was implemented in the force field COSMOS-NMR and applied to the histidine-tryptophan-complex of the transmembrane domain of the M2 protein channel of influenza A virus. Our calculations show that moderate changes of side chain torsion angle χ1 and small variations of χ2 of Trp-41 are necessary to switch from the inactivated into the activated state; and a rough two-side jump model of His-37 is supported for proton gating in accordance with a flipping mechanism.

Increasing Protein Charge State When Using Laser Electrospray Mass Spectrometry

Science.gov (United States)

Karki, Santosh; Flanigan, Paul M.; Perez, Johnny J.; Archer, Jieutonne J.; Levis, Robert J.

2015-05-01

Femtosecond (fs) laser vaporization is used to transfer cytochrome c, myoglobin, lysozyme, and ubiquitin from the condensed phase into an electrospray (ES) plume consisting of a mixture of a supercharging reagent, m-nitrobenzyl alcohol ( m-NBA), and trifluoroacetic acid (TFA), acetic acid (AA), or formic acid (FA). Interaction of acid-sensitive proteins like cytochrome c and myoglobin with the highly charged ES droplets resulted in a shift to higher charge states in comparison with acid-stable proteins like lysozyme and ubiquitin. Laser electrospray mass spectrometry (LEMS) measurements showed an increase in both the average charge states (Zavg) and the charge state with maximum intensity (Zmode) for acid-sensitive proteins compared with conventional electrospray ionization mass spectrometry (ESI-MS) under equivalent solvent conditions. A marked increase in ion abundance of higher charge states was observed for LEMS in comparison with conventional electrospray for cytochrome c (ranging from 19+ to 21+ versus 13+ to 16+) and myoglobin (ranging from 19+ to 26+ versus 18+ to 21+) using an ES solution containing m-NBA and TFA. LEMS measurements as a function of electrospray flow rate yielded increasing charge states with decreasing flow rates for cytochrome c and myoglobin.

17 CFR 229.503 - (Item 503) Prospectus summary, risk factors, and ratio of earnings to fixed charges.

Science.gov (United States)

2010-04-01

... ratio of earnings to fixed charges. The registrant must furnish this information in plain English. See... you do not caption it as a summary, you still must provide that information in plain English... discussion of the most significant factors that make the offering speculative or risky. This discussion must...

An albumin-fixed membrane for the removal of protein-bound toxins

International Nuclear Information System (INIS)

Ge Dongtao; Wu Dewang; Shi Wei; Ma Yuanyuan; Tian Xiangdong; Liang Pengfei; Zhang Qiqing

2006-01-01

Established methods for kidney dialysis do not work for liver failure because kidney dialysis removes only water-soluble toxins, while the liver normally removes albumin-bound toxins. In the present study, a polysulfone dialysis membrane with a -OH reactive group was prepared by hydrolyzing the chloromethylated polysulfone membrane, and the bovine serum albumin molecules were fixed into the membrane with 1,1'-carbonyldiimidazole activation. The content of albumin of the albumin-fixed membrane was 121.3 mg (g membrane) -1 . The albumin-fixed dialysis membranes were used to remove protein-bound toxins, bilirubin, from the bilirubin-albumin solution. The transfer rate of bilirubin of the albumin-fixed membrane was obviously higher compared to the normal dialysis membrane. The clearance of bilirubin with the albumin-fixed membrane was 49.8%. The albumin-fixed membrane can easily be regenerated by the bovine serum albumin and NaOH solution. Regeneration of the membrane suggested good mechanical and chemical stability, as well as good clearance of bilirubin. In addition, the effects of membrane thickness and bilirubin initial concentration on the removal of bilirubin were discussed

Charge Segregation and Low Hydrophobicity Are Key Features of Ribosomal Proteins from Different Organisms*

Science.gov (United States)

Fedyukina, Daria V.; Jennaro, Theodore S.; Cavagnero, Silvia

2014-01-01

Ribosomes are large and highly charged macromolecular complexes consisting of RNA and proteins. Here, we address the electrostatic and nonpolar properties of ribosomal proteins that are important for ribosome assembly and interaction with other cellular components and may influence protein folding on the ribosome. We examined 50 S ribosomal subunits from 10 species and found a clear distinction between the net charge of ribosomal proteins from halophilic and non-halophilic organisms. We found that ∼67% ribosomal proteins from halophiles are negatively charged, whereas only up to ∼15% of ribosomal proteins from non-halophiles share this property. Conversely, hydrophobicity tends to be lower for ribosomal proteins from halophiles than for the corresponding proteins from non-halophiles. Importantly, the surface electrostatic potential of ribosomal proteins from all organisms, especially halophiles, has distinct positive and negative regions across all the examined species. Positively and negatively charged residues of ribosomal proteins tend to be clustered in buried and solvent-exposed regions, respectively. Hence, the majority of ribosomal proteins is characterized by a significant degree of intramolecular charge segregation, regardless of the organism of origin. This key property enables the ribosome to accommodate proteins within its complex scaffold regardless of their overall net charge. PMID:24398678

Modified Protein Improves Vitiligo Symptoms in Mice

Science.gov (United States)

... Vitiligo Symptoms in Mice Spotlight on Research Modified Protein Improves Vitiligo Symptoms in Mice By Colleen Labbe, ... D., Ph.D., Rush University. Altering a key protein involved in the development of vitiligo may protect ...

Change in carrier type in high-k gate carbon nanotube field-effect transistors by interface fixed charges

International Nuclear Information System (INIS)

Moriyama, N; Ohno, Y; Kitamura, T; Kishimoto, S; Mizutani, T

2010-01-01

We study the phenomenon of change in carrier type in carbon nanotube field-effect transistors (CNFETs) caused by the atomic layer deposition (ALD) of a HfO 2 gate insulator. When a HfO 2 layer is deposited on a CNFET, the type of carrier changes from p-type to n-type. The so-obtained n-type device has good performance and stability in air. The conductivity of such a device with a channel length of 0.7 μm is 11% of the quantum conductance 4e 2 /h. The contact resistance for electron current is estimated to be 14 kΩ. The n-type conduction of this CNFET is maintained for more than 100 days. The change in carrier type is attributed to positive fixed charges introduced at the interface between the HfO 2 and SiO 2 layers. We also propose a novel technique to control the type of conduction by utilizing interface fixed charges; this technique is compatible with Si CMOS process technology.

A Novel, In-solution Separation of Endogenous Cardiac Sarcomeric Proteins and Identification of Distinct Charged Variants of Regulatory Light Chain*

Science.gov (United States)

Scruggs, Sarah B.; Reisdorph, Rick; Armstrong, Mike L.; Warren, Chad M.; Reisdorph, Nichole; Solaro, R. John; Buttrick, Peter M.

2010-01-01

The molecular conformation of the cardiac myosin motor is modulated by intermolecular interactions among the heavy chain, the light chains, myosin binding protein-C, and titin and is governed by post-translational modifications (PTMs). In-gel digestion followed by LC/MS/MS has classically been applied to identify cardiac sarcomeric PTMs; however, this approach is limited by protein size, pI, and difficulties in peptide extraction. We report a solution-based work flow for global separation of endogenous cardiac sarcomeric proteins with a focus on the regulatory light chain (RLC) in which specific sites of phosphorylation have been unclear. Subcellular fractionation followed by OFFGEL electrophoresis resulted in isolation of endogenous charge variants of sarcomeric proteins, including regulatory and essential light chains, myosin heavy chain, and myosin-binding protein-C of the thick filament. Further purification of RLC using reverse-phase HPLC separation and UV detection enriched for RLC PTMs at the intact protein level and provided a stoichiometric and quantitative assessment of endogenous RLC charge variants. Digestion and subsequent LC/MS/MS unequivocally identified that the endogenous charge variants of cardiac RLC focused in unique OFFGEL electrophoresis fractions were unphosphorylated (78.8%), singly phosphorylated (18.1%), and doubly phosphorylated (3.1%) RLC. The novel aspects of this study are that 1) milligram amounts of endogenous cardiac sarcomeric subproteome were focused with resolution comparable with two-dimensional electrophoresis, 2) separation and quantification of post-translationally modified variants were achieved at the intact protein level, 3) separation of intact high molecular weight thick filament proteins was achieved in solution, and 4) endogenous charge variants of RLC were separated; a novel doubly phosphorylated form was identified in mouse, and singly phosphorylated, singly deamidated, and deamidated/phosphorylated forms were

Protein extraction from methanol fixed paraffin embedded tissue blocks: A new possibility using cell blocks

Science.gov (United States)

Kokkat, Theresa J.; McGarvey, Diane; Patel, Miral S.; Tieniber, Andrew D.; LiVolsi, Virginia A.; Baloch, Zubair W.

2013-01-01

Background: Methanol fixed and paraffin embedded (MFPE) cellblocks are an essential cytology preparation. However, MFPE cellblocks often contain limited material and their relatively small size has caused them to be overlooked in biomarker discovery. Advances in the field of molecular biotechnology have made it possible to extract proteins from formalin fixed and paraffin embedded (FFPE) tissue blocks. In contrast, there are no established methods for extracting proteins from MFPE cellblocks. We investigated commonly available CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate) buffer, as well as two commercially available Qiagen® kits and compared their effectiveness on MFPE tissue for protein yields. Materials and Methods: MFPE blocks were made by Cellient™ automated system using human tissue specimens from normal and malignant specimens collected in ThinPrep™ Vials. Protein was extracted from Cellient-methanol fixed and paraffin embedded blocks with CHAPS buffer method as well as FFPE and Mammalian Qiagen® kits. Results: Comparison of protein yields demonstrated the effectiveness of various protein extraction methods on MFPE cellblocks. Conclusion: In the current era of minimally invasive techniques to obtain minimal amount of tissue for diagnostic and prognostic purposes, the use of commercial and lab made buffer on low weight MFPE scrapings obtained by Cellient® processor opens new possibilities for protein biomarker research. PMID:24403950

Bioreducible poly(amidoamine)s with charge-reversel properties for intracellular protein delivery

NARCIS (Netherlands)

Coué, G.M.J.P.C.; Engbersen, Johannes F.J.; Hennink, W.E.; Engbersen, J.F.J.

2010-01-01

An effective intracellular protein delivery system was developed using bioreducible disulfide-containing poly(amidoamine)s with negatively charged citraconic side groups that can give charge-reversal upon pH decrease. These water-soluble and linear polymers efficiently self-assemble with proteins

Artificial immune system and sheep flock algorithms for two-stage fixed-charge transportation problem

DEFF Research Database (Denmark)

Kannan, Devika; Govindan, Kannan; Soleimani, Hamed

2014-01-01

In this paper, we cope with a two-stage distribution planning problem of supply chain regarding fixed charges. The focus of the paper is on developing efficient solution methodologies of the selected NP-hard problem. Based on computational limitations, common exact and approximation solution...... approaches are unable to solve real-world instances of such NP-hard problems in a reasonable time. These approaches involve cumbersome computational steps in real-size cases. In order to solve the mixed integer linear programming model, we develop an artificial immune system and a sheep flock algorithm...

Theoretical model for the detection of charged proteins with a silicon-on-insulator sensor

International Nuclear Information System (INIS)

Birner, S; Uhl, C; Bayer, M; Vogl, P

2008-01-01

For a bio-sensor device based on a silicon-on-insulator structure, we calculate the sensitivity to specific charge distributions in the electrolyte solution that arise from protein binding to the semiconductor surface. This surface is bio-functionalized with a lipid layer so that proteins can specifically bind to the headgroups of the lipids on the surface. We consider charged proteins such as the green fluorescent protein (GFP) and artificial proteins that consist of a variable number of aspartic acids. Specifically, we calculate self-consistently the spatial charge and electrostatic potential distributions for different ion concentrations in the electrolyte. We fully take into account the quantum mechanical charge density in the semiconductor. We determine the potential change at the binding sites as a function of protein charge and ionic strength. Comparison with experiment is generally very good. Furthermore, we demonstrate the superiority of the full Poisson-Boltzmann equation by comparing its results to the simplified Debye-Hueckel approximation

Polysaccharide charge density regulating protein adsorption to air/water interfaces by protein/polysaccharide complex formation

NARCIS (Netherlands)

Ganzevles, R.A.; Kosters, H.; Vliet, T. van; Stuart, M.A.C.; Jongh, H.H.J. de

2007-01-01

Because the formation of protein/polysaccharide complexes is dominated by electrostatic interaction, polysaccharide charge density is expected to play a major role in the adsorption behavior of the complexes. In this study, pullulan (a non-charged polysaccharide) carboxylated to four different

Superficial Collagen Fibril Modulus and Pericellular Fixed Charge Density Modulate Chondrocyte Volumetric Behaviour in Early Osteoarthritis

Directory of Open Access Journals (Sweden)

Petri Tanska

2013-01-01

Full Text Available The aim of this study was to investigate if the experimentally detected altered chondrocyte volumetric behavior in early osteoarthritis can be explained by changes in the extracellular and pericellular matrix properties of cartilage. Based on our own experimental tests and the literature, the structural and mechanical parameters for normal and osteoarthritic cartilage were implemented into a multiscale fibril-reinforced poroelastic swelling model. Model simulations were compared with experimentally observed cell volume changes in mechanically loaded cartilage, obtained from anterior cruciate ligament transected rabbit knees. We found that the cell volume increased by 7% in the osteoarthritic cartilage model following mechanical loading of the tissue. In contrast, the cell volume decreased by 4% in normal cartilage model. These findings were consistent with the experimental results. Increased local transversal tissue strain due to the reduced collagen fibril stiffness accompanied with the reduced fixed charge density of the pericellular matrix could increase the cell volume up to 12%. These findings suggest that the increase in the cell volume in mechanically loaded osteoarthritic cartilage is primarily explained by the reduction in the pericellular fixed charge density, while the superficial collagen fibril stiffness is suggested to contribute secondarily to the cell volume behavior.

Patients with polymyositis show changes in muscle protein charges

DEFF Research Database (Denmark)

Bartels, E M; Jacobsen, Søren; Rasmussen, L

1989-01-01

Polymyositis (PM) appears with indolent proximal muscle weakness and is an inflammatory disease with breakdown of muscle cells. In our study the protein charge concentrations of the contractile proteins in the A and I bands were determined, applying a microelectrode technique. Patients with PM sh...

Substantial conformational change mediated by charge-triad residues of the death effector domain in protein-protein interactions.

Directory of Open Access Journals (Sweden)

Edward C Twomey

Full Text Available Protein conformational changes are commonly associated with the formation of protein complexes. The non-catalytic death effector domains (DEDs mediate protein-protein interactions in a variety of cellular processes, including apoptosis, proliferation and migration, and glucose metabolism. Here, using NMR residual dipolar coupling (RDC data, we report a conformational change in the DED of the phosphoprotein enriched in astrocytes, 15 kDa (PEA-15 protein in the complex with a mitogen-activated protein (MAP kinase, extracellular regulated kinase 2 (ERK2, which is essential in regulating ERK2 cellular distribution and function in cell proliferation and migration. The most significant conformational change in PEA-15 happens at helices α2, α3, and α4, which also possess the highest flexibility among the six-helix bundle of the DED. This crucial conformational change is modulated by the D/E-RxDL charge-triad motif, one of the prominent structural features of DEDs, together with a number of other electrostatic and hydrogen bonding interactions on the protein surface. Charge-triad motif promotes the optimal orientation of key residues and expands the binding interface to accommodate protein-protein interactions. However, the charge-triad residues are not directly involved in the binding interface between PEA-15 and ERK2.

A batch and fixed bed column study for fluorescein removal using chitosan modified by epichlorohydrin.

Science.gov (United States)

Silva, P M O; Francisco, J E; Cajé, J C M; Cassella, R J; Pacheco, W F

2018-01-02

This study evaluates the feasibility of commercial chitosan (CQ) and modified chitosan (MQ) by epichlorohydrin to be used as a solid phase to remove fluorescein (FSC) from aqueous solutions by two different approaches: in batch and on a fixed column bed. For the batch study, all parameters that influence sorption capacity were evaluated, such as: pH, mass, ionic strength, temperature and time of contact. In the optimized condition, 75% removal was obtained for FSC using CQ, while the modification allowed an increase up to 99%, as well as an increase in the stability of the polymer. In the fixed column bed study, the influence of all the parameters was evaluated through breakthrough curves, and the thermodynamics parameters of each approach were obtained. The results of these studies demonstrate that the modification with epichlorohydrin enhanced the sorptive properties (from 35% to 95% in fixed bed experiments) and the polymer stability (making it insoluble), making it suitable to be used in wastewater treatment.

Role of Charge Regulation and Size Polydispersity in Nanoparticle Encapsulation by Viral Coat Proteins

NARCIS (Netherlands)

Kusters, Remy; Lin, Hsiang-Ku; Zandi, Roya; Tsvetkova, Irina; Dragnea, Bogdan; van der Schoot, Paul

2015-01-01

Nanoparticles can be encapsulated by virus coat proteins if their surfaces are functionalized to acquire a sufficiently large negative charge. A minimal surface charge is required to overcome (i) repulsive interactions between the positively charged RNA-binding domains on the proteins and (ii) the

Sequence charge decoration dictates coil-globule transition in intrinsically disordered proteins.

Science.gov (United States)

Firman, Taylor; Ghosh, Kingshuk

2018-03-28

We present an analytical theory to compute conformations of heteropolymers-applicable to describe disordered proteins-as a function of temperature and charge sequence. The theory describes coil-globule transition for a given protein sequence when temperature is varied and has been benchmarked against the all-atom Monte Carlo simulation (using CAMPARI) of intrinsically disordered proteins (IDPs). In addition, the model quantitatively shows how subtle alterations of charge placement in the primary sequence-while maintaining the same charge composition-can lead to significant changes in conformation, even as drastic as a coil (swelled above a purely random coil) to globule (collapsed below a random coil) and vice versa. The theory provides insights on how to control (enhance or suppress) these changes by tuning the temperature (or solution condition) and charge decoration. As an application, we predict the distribution of conformations (at room temperature) of all naturally occurring IDPs in the DisProt database and notice significant size variation even among IDPs with a similar composition of positive and negative charges. Based on this, we provide a new diagram-of-states delineating the sequence-conformation relation for proteins in the DisProt database. Next, we study the effect of post-translational modification, e.g., phosphorylation, on IDP conformations. Modifications as little as two-site phosphorylation can significantly alter the size of an IDP with everything else being constant (temperature, salt concentration, etc.). However, not all possible modification sites have the same effect on protein conformations; there are certain "hot spots" that can cause maximal change in conformation. The location of these "hot spots" in the parent sequence can readily be identified by using a sequence charge decoration metric originally introduced by Sawle and Ghosh. The ability of our model to predict conformations (both expanded and collapsed states) of IDPs at a high

Fixed poles in electromagnetic processes and modification of Adler's neutrino sum rule due to quark anomalous magnetic moment

International Nuclear Information System (INIS)

Khare, A.

1975-01-01

We show that Adler's sum rule for neutrino scattering and Bjorken's inequality for electron-proton scattering are modified if quark has finite anomalous magnetic moment ksub(q). We also show that if ksub(q) is nonzero, there exist fixed poles in spin-flip Compton scattering as well as in charged pion photoproduction. (auth.)

Mixed-mode reversed phase/positively charged repulsion chromatography for intact protein separation.

Science.gov (United States)

Ding, Ling; Guo, Zhimou; Hu, Zhuo; Liang, Xinmiao

2017-05-10

A mixed-mode reversed phase/positively charged repulsion stationary phase C8PN composed of octyl and amino group has been developed for separation of intact protein. Before the separation of proteins, a set of probe compounds were employed to evaluate the chromatographic properties of C8PN, demonstrating typical reversed phase/positively charged repulsion interaction on this stationary phase as estimated. Then the new C8PN stationary phase was used to separate a standard protein mixture on the reversed phase mode. Compared with a commercial C4 stationary phase, it showed different selectivity for some proteins. In order to better understand the properties of C8PN, the effect of acetonitrile content was investigated based on retention equation. Higher values of the equation parameters on C8PN demonstrated that the protein retentions were more sensitive to the change of acetonitrile content. Besides, the influences of buffer salt additives on the protein retentions were also studied. The retention factors of the proteins got larger with the increase of buffer salt concentration, which confirmed the positively charged repulsion interaction on the column. Finally, the C8PN was further applied to separate oxidized- and reduced- forms of Recombinant Human Growth Hormone. Our study indicated the advantages and application potential of mixed-mode reversed phase/positively charged repulsion stationary phase for intact protein separation. Copyright © 2017 Elsevier B.V. All rights reserved.

Effect of incorporation of nitrogen atoms in Al2O3 gate dielectric of wide-bandgap-semiconductor MOSFET on gate leakage current and negative fixed charge

Science.gov (United States)

Kojima, Eiji; Chokawa, Kenta; Shirakawa, Hiroki; Araidai, Masaaki; Hosoi, Takuji; Watanabe, Heiji; Shiraishi, Kenji

2018-06-01

We performed first-principle calculations to investigate the effect of incorporation of N atoms into Al2O3 gate dielectrics. Our calculations show that the defect levels generated by VO in Al2O3 are the origin of the stress-induced gate leakage current and that VOVAl complexes in Al2O3 cause negative fixed charge. We revealed that the incorporation of N atoms into Al2O3 eliminates the VO defect levels, reducing the stress-induced gate leakage current. Moreover, this suppresses the formation of negatively charged VOVAl complexes. Therefore, AlON can reduce both stress-induced gate leakage current and negative fixed charge in wide-bandgap-semiconductor MOSFETs.

Novel Highly Sensitive Protein Sensors Based on Tapered Optical Fibres Modified with Au-Based Nanocoatings

Directory of Open Access Journals (Sweden)

Aitor Urrutia

2016-01-01

Full Text Available Novel protein sensors based on tapered optical fibres modified with Au coatings deposited using two different procedures are proposed. Au-based coatings are deposited onto a nonadiabatic tapered optical fibre using (i a novel facile method composed of layer-by-layer deposition consisting of polycation (poly(allylamine hydrochloride, PAH and negatively charged SiO2 nanoparticles (NPs followed by the deposition of the charged Au NPs and (ii the sputtering technique. The Au NPs and Au thin film surfaces are then modified with biotin in order to bind streptavidin (SV molecules and detect them. The sensing principle is based on the sensitivity of the transmission spectrum of the device to changes in the refractive index of the coatings induced by the SV binding to the biotin. Both sensors showed high sensitivity to SV, with the lowest measured concentration levels below 2.5 nM. The calculated binding constant for the biotin-SV pair was 2.2×10-11 M−1 when a tapered fibre modified with the LbL method was used, with a limit of detection (LoD of 271 pM. The sensor formed using sputtering had a binding constant of 1.01×10-10 M−1 with a LoD of 806 pM. These new structures and their simple fabrication technique could be used to develop other biosensors.

A charge-dependent mechanism is responsible for the dynamic accumulation of proteins inside nucleoli.

Science.gov (United States)

Musinova, Yana R; Kananykhina, Eugenia Y; Potashnikova, Daria M; Lisitsyna, Olga M; Sheval, Eugene V

2015-01-01

The majority of known nucleolar proteins are freely exchanged between the nucleolus and the surrounding nucleoplasm. One way proteins are retained in the nucleoli is by the presence of specific amino acid sequences, namely nucleolar localization signals (NoLSs). The mechanism by which NoLSs retain proteins inside the nucleoli is still unclear. Here, we present data showing that the charge-dependent (electrostatic) interactions of NoLSs with nucleolar components lead to nucleolar accumulation as follows: (i) known NoLSs are enriched in positively charged amino acids, but the NoLS structure is highly heterogeneous, and it is not possible to identify a consensus sequence for this type of signal; (ii) in two analyzed proteins (NF-κB-inducing kinase and HIV-1 Tat), the NoLS corresponds to a region that is enriched for positively charged amino acid residues; substituting charged amino acids with non-charged ones reduced the nucleolar accumulation in proportion to the charge reduction, and nucleolar accumulation efficiency was strongly correlated with the predicted charge of the tested sequences; and (iii) sequences containing only lysine or arginine residues (which were referred to as imitative NoLSs, or iNoLSs) are accumulated in the nucleoli in a charge-dependent manner. The results of experiments with iNoLSs suggested that charge-dependent accumulation inside the nucleoli was dependent on interactions with nucleolar RNAs. The results of this work are consistent with the hypothesis that nucleolar protein accumulation by NoLSs can be determined by the electrostatic interaction of positively charged regions with nucleolar RNAs rather than by any sequence-specific mechanism. Copyright © 2014 Elsevier B.V. All rights reserved.

Catalytic Micromotors Moving Near Polyelectrolyte-Modified Substrates: The Roles of Surface Charges, Morphology, and Released Ions.

Science.gov (United States)

Wei, Mengshi; Zhou, Chao; Tang, Jinyao; Wang, Wei

2018-01-24

Synthetic microswimmers, or micromotors, are finding potential uses in a wide range of applications, most of which involve boundaries. However, subtle yet important effects beyond physical confinement on the motor dynamics remain less understood. In this letter, glass substrates were functionalized with positively and negatively charged polyelectrolytes, and the dynamics of micromotors moving close to the modified surfaces was examined. Using acoustic levitation and numerical simulation, we reveal how the speed of a chemically propelled micromotor slows down significantly near a polyelectrolyte-modified surface by the combined effects of surface charges, surface morphology, and ions released from the films.

Modelling of electric tree progression due to space charge modified fields

International Nuclear Information System (INIS)

Seralathan, K E; Mahajan, A; Gupta, Nandini

2008-01-01

Tree initiation and growth require localized field enhancement that results in material erosion and formation of tree channels. Tree progression is linked to partial discharges within the tree tubules, characterized by recurrent periods of activity followed by quiescent states. Charge builds up across the non-conducting tree channels during the inactive regime, and discharge follows. In this work, the role of the space charge modified field during the non-discharging regime in deciding the site of subsequent discharges and thereby shaping tree structures is studied. A simple stochastic model was developed, in order to understand the respective effects of charges trapped on the walls of tree tubules, at channel tips, or in the volume of the dielectric. While some charge distributions are seen to arrest tree growth, others encourage axial growth towards the other electrode, and some aid in producing bushy trees clustered around the needle tip. The effect of carbon deposition within tree channels, making them effectively conducting, was also investigated. The insights gained from the simulations were successfully used to explain tree growth in the laboratory under high- and low-field conditions

A modified Poisson-Boltzmann equation applied to protein adsorption.

Science.gov (United States)

Gama, Marlon de Souza; Santos, Mirella Simões; Lima, Eduardo Rocha de Almeida; Tavares, Frederico Wanderley; Barreto, Amaro Gomes Barreto

2018-01-05

Ion-exchange chromatography has been widely used as a standard process in purification and analysis of protein, based on the electrostatic interaction between the protein and the stationary phase. Through the years, several approaches are used to improve the thermodynamic description of colloidal particle-surface interaction systems, however there are still a lot of gaps specifically when describing the behavior of protein adsorption. Here, we present an improved methodology for predicting the adsorption equilibrium constant by solving the modified Poisson-Boltzmann (PB) equation in bispherical coordinates. By including dispersion interactions between ions and protein, and between ions and surface, the modified PB equation used can describe the Hofmeister effects. We solve the modified Poisson-Boltzmann equation to calculate the protein-surface potential of mean force, treated as spherical colloid-plate system, as a function of process variables. From the potential of mean force, the Henry constants of adsorption, for different proteins and surfaces, are calculated as a function of pH, salt concentration, salt type, and temperature. The obtained Henry constants are compared with experimental data for several isotherms showing excellent agreement. We have also performed a sensitivity analysis to verify the behavior of different kind of salts and the Hofmeister effects. Copyright © 2017 Elsevier B.V. All rights reserved.

4-Hydroxyhexenal- and 4-Hydroxynonenal-Modified Proteins in Pterygia

Directory of Open Access Journals (Sweden)

Ichiya Sano

2013-01-01

Full Text Available Oxidative stress has been suspected of contributing to the pathogenesis of pterygia. We evaluated the immunohistochemical localization of the markers of oxidative stress, that is, the proteins modified by 4-hydroxyhexenal (4-HHE and 4-hydroxynonenal (4-HNE, which are reactive aldehydes derived from nonenzymatic oxidation of n-3 and n-6 polyunsaturated fatty acids, respectively. In the pterygial head, labeling of 4-HHE- and 4-HNE-modified proteins was prominent in the nuclei and cytosol of the epithelium. In the pterygial body, strong labeling was observed in the nuclei and cytosol of the epithelium and proliferating subepithelial connective tissue. In normal conjunctival specimens, only trace immunoreactivity of both proteins was observed in the epithelial and stromal layers. Exposures of ultraviolet (330 nm, 48.32 ± 0.55 J/cm2 or blue light (400 nm, 293.0 ± 2.0 J/cm2 to rat eyes enhanced labeling of 4-HHE- and 4-HNE-modified proteins in the nuclei of conjunctival epithelium. Protein modifications by biologically active aldehydes are a molecular event involved in the development of pterygia.

Improved protein extraction and protein identification from archival formalin-fixed paraffin-embedded human aortas.

Science.gov (United States)

Fu, Zongming; Yan, Kun; Rosenberg, Avraham; Jin, Zhicheng; Crain, Barbara; Athas, Grace; Heide, Richard S Vander; Howard, Timothy; Everett, Allen D; Herrington, David; Van Eyk, Jennifer E

2013-04-01

Evaluate combination of heat and elevated pressure to enhance protein extraction and quality of formalin-fixed (FF), and FF paraffin-embedded (FFPE) aorta for proteomics. Proteins were extracted from fresh frozen aorta at room temperature (RT). FF and FFPE aortas (3 months and 15 years) were extracted at RT, heat alone, or a combination of heat and high pressure. Protein yields were compared, and digested peptides from the extracts were analyzed with MS. Combined heat and elevated pressure increased protein yield from human FF or FFPE aorta compared to matched tissues with heat alone (1.5-fold) or at RT (8.3-fold), resulting in more proteins identified and with more sequence coverage. The length of storage did adversely affect the quality of proteins from FF tissue. For long-term storage, aorta was preserved better with FFPE than FF alone. Periostin and MGF-E8 were demonstrated suitable for MRM assays from FFPE aorta. Combination of heat and high pressure is an effective method to extract proteins from FFPE aorta for downstream proteomics. This method opens the possibility for use of archival and often rare FFPE aortas and possibly other tissues available to proteomics for biomarker discovery and quantification. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Sequence charge decoration dictates coil-globule transition in intrinsically disordered proteins

Science.gov (United States)

Firman, Taylor; Ghosh, Kingshuk

2018-03-01

We present an analytical theory to compute conformations of heteropolymers—applicable to describe disordered proteins—as a function of temperature and charge sequence. The theory describes coil-globule transition for a given protein sequence when temperature is varied and has been benchmarked against the all-atom Monte Carlo simulation (using CAMPARI) of intrinsically disordered proteins (IDPs). In addition, the model quantitatively shows how subtle alterations of charge placement in the primary sequence—while maintaining the same charge composition—can lead to significant changes in conformation, even as drastic as a coil (swelled above a purely random coil) to globule (collapsed below a random coil) and vice versa. The theory provides insights on how to control (enhance or suppress) these changes by tuning the temperature (or solution condition) and charge decoration. As an application, we predict the distribution of conformations (at room temperature) of all naturally occurring IDPs in the DisProt database and notice significant size variation even among IDPs with a similar composition of positive and negative charges. Based on this, we provide a new diagram-of-states delineating the sequence-conformation relation for proteins in the DisProt database. Next, we study the effect of post-translational modification, e.g., phosphorylation, on IDP conformations. Modifications as little as two-site phosphorylation can significantly alter the size of an IDP with everything else being constant (temperature, salt concentration, etc.). However, not all possible modification sites have the same effect on protein conformations; there are certain "hot spots" that can cause maximal change in conformation. The location of these "hot spots" in the parent sequence can readily be identified by using a sequence charge decoration metric originally introduced by Sawle and Ghosh. The ability of our model to predict conformations (both expanded and collapsed states) of IDPs at

Ion selection of charge-modified large nanopores in a graphene sheet

Science.gov (United States)

Zhao, Shijun; Xue, Jianming; Kang, Wei

2013-09-01

Water desalination becomes an increasingly important approach for clean water supply to meet the rapidly growing demand of population boost, industrialization, and urbanization. The main challenge in current desalination technologies lies in the reduction of energy consumption and economic costs. Here, we propose to use charged nanopores drilled in a graphene sheet as ion exchange membranes to promote the efficiency and capacity of desalination systems. Using molecular dynamics simulations, we investigate the selective ion transport behavior of electric-field-driven KCl electrolyte solution through charge modified graphene nanopores. Our results reveal that the presence of negative charges at the edge of graphene nanopore can remarkably impede the passage of Cl- while enhance the transport of K+, which is an indication of ion selectivity for electrolytes. We further demonstrate that this selectivity is dependent on the pore size and total charge number assigned at the nanopore edge. By adjusting the nanopore diameter and electric charge on the graphene nanopore, a nearly complete rejection of Cl- can be realized. The electrical resistance of nanoporous graphene, which is a key parameter to evaluate the performance of ion exchange membranes, is found two orders of magnitude lower than commercially used membranes. Our results thus suggest that graphene nanopores are promising candidates to be used in electrodialysis technology for water desalinations with a high permselectivity.

Myofibrillar protein oxidation affects filament charges, aggregation and water-holding

NARCIS (Netherlands)

Bao, Yulong; Boeren, Sjef; Ertbjerg, Per

2018-01-01

Hypochlorous acid (HClO) is a strong oxidant that is able to mediate protein oxidation. In order to study the effect of oxidation on charges, aggregation and water-holding of myofibrillar proteins, extracted myofibrils were oxidized by incubation with different concentrations of HClO (0, 1, 5,

Determination of gas phase protein ion densities via ion mobility analysis with charge reduction.

Science.gov (United States)

Maisser, Anne; Premnath, Vinay; Ghosh, Abhimanyu; Nguyen, Tuan Anh; Attoui, Michel; Hogan, Christopher J

2011-12-28

We use a charge reduction electrospray (ESI) source and subsequent ion mobility analysis with a differential mobility analyzer (DMA, with detection via both a Faraday cage electrometer and a condensation particle counter) to infer the densities of single and multiprotein ions of cytochrome C, lysozyme, myoglobin, ovalbumin, and bovine serum albumin produced from non-denaturing (20 mM aqueous ammonium acetate) and denaturing (1 : 49.5 : 49.5, formic acid : methanol : water) ESI. Charge reduction is achieved through use of a Po-210 radioactive source, which generates roughly equal concentrations of positive and negative ions. Ions produced by the source collide with and reduce the charge on ESI generated drops, preventing Coulombic fissions, and unlike typical protein ESI, leading to gas-phase protein ions with +1 to +3 excess charges. Therefore, charge reduction serves to effectively mitigate any role that Coulombic stretching may play on the structure of the gas phase ions. Density inference is made via determination of the mobility diameter, and correspondingly the spherical equivalent protein volume. Through this approach it is found that for both non-denaturing and denaturing ESI-generated ions, gas-phase protein ions are relatively compact, with average densities of 0.97 g cm(-3) and 0.86 g cm(-3), respectively. Ions from non-denaturing ESI are found to be slightly more compact than predicted from the protein crystal structures, suggesting that low charge state protein ions in the gas phase are slightly denser than their solution conformations. While a slight difference is detected between the ions produced with non-denaturing and denaturing ESI, the denatured ions are found to be much more dense than those examined previously by drift tube mobility analysis, in which charge reduction was not employed. This indicates that Coulombic stretching is typically what leads to non-compact ions in the gas-phase, and suggests that for gas phase

Protein adsorption resistance of PVP-modified polyurethane film prepared by surface-initiated atom transfer radical polymerization

International Nuclear Information System (INIS)

Yuan, Huihui; Qian, Bin; Zhang, Wei; Lan, Minbo

2016-01-01

Highlights: • Antifouling PVP brushes were successfully grafted on PU films by SI-ATRP. • The effect of polymerization time on surface property and topography was studied. • Hydrophilicity and protein fouling resistance of PVP–PU films were greatly promoted. • Competitive adsorption of three proteins on PVP–PU films was evaluated. - Abstract: An anti-fouling surface of polyurethane (PU) film grafted with Poly(N-vinylpyrrolidone) (PVP) was prepared through surface-initiated atom transfer radical polymerization (SI-ATRP). And the polymerization time was investigated to obtain PU films with PVP brushes of different lengths. The surface properties and protein adsorption of modified PU films were evaluated. The results showed that the hydrophilicity of PU–PVP films were improved with the increase of polymerization time, which was not positive correlation with the surface roughness due to the brush structure. Additionally, the protein resistance performance was promoted when prolonging the polymerization time. The best antifouling PU–PVP (6.0 h) film reduced the adsoption level of bovine serum albumin (BSA), lysozyme (LYS), and brovin serum fibrinogen (BFG) by 93.4%, 68.3%, 85.6%, respectively, compared to the unmodified PU film. The competitive adsorption of three proteins indicated that LYS preferentially adsorbed on the modified PU film, while BFG had the lowest adsorption selectivity. And the amount of BFG on PU–PVP (6.0 h) film reduced greatly to 0.08 μg/cm"2, which was almost one-tenth of its adsorption from the single-protein system. Presented results suggested that both hydrophilicity and surface roughness might be the important factors in all cases of protein adsorption, and the competitive or selective adsorption might be related to the size of the proteins, especially on the non-charged films.

Protein adsorption resistance of PVP-modified polyurethane film prepared by surface-initiated atom transfer radical polymerization

Energy Technology Data Exchange (ETDEWEB)

Yuan, Huihui; Qian, Bin; Zhang, Wei [Shanghai Key Laboratory of Functional Materials Chemistry and Research Center of Analysis and Test, East China University of Science and Technology, Shanghai 200237 (China); Lan, Minbo, E-mail: minbolan@ecust.edu.cn [Shanghai Key Laboratory of Functional Materials Chemistry and Research Center of Analysis and Test, East China University of Science and Technology, Shanghai 200237 (China); State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237 (China)

2016-02-15

Highlights: • Antifouling PVP brushes were successfully grafted on PU films by SI-ATRP. • The effect of polymerization time on surface property and topography was studied. • Hydrophilicity and protein fouling resistance of PVP–PU films were greatly promoted. • Competitive adsorption of three proteins on PVP–PU films was evaluated. - Abstract: An anti-fouling surface of polyurethane (PU) film grafted with Poly(N-vinylpyrrolidone) (PVP) was prepared through surface-initiated atom transfer radical polymerization (SI-ATRP). And the polymerization time was investigated to obtain PU films with PVP brushes of different lengths. The surface properties and protein adsorption of modified PU films were evaluated. The results showed that the hydrophilicity of PU–PVP films were improved with the increase of polymerization time, which was not positive correlation with the surface roughness due to the brush structure. Additionally, the protein resistance performance was promoted when prolonging the polymerization time. The best antifouling PU–PVP (6.0 h) film reduced the adsoption level of bovine serum albumin (BSA), lysozyme (LYS), and brovin serum fibrinogen (BFG) by 93.4%, 68.3%, 85.6%, respectively, compared to the unmodified PU film. The competitive adsorption of three proteins indicated that LYS preferentially adsorbed on the modified PU film, while BFG had the lowest adsorption selectivity. And the amount of BFG on PU–PVP (6.0 h) film reduced greatly to 0.08 μg/cm{sup 2}, which was almost one-tenth of its adsorption from the single-protein system. Presented results suggested that both hydrophilicity and surface roughness might be the important factors in all cases of protein adsorption, and the competitive or selective adsorption might be related to the size of the proteins, especially on the non-charged films.

Solving the Single-Sink, Fixed-Charge, Multiple-Choice Transportation Problem by Dynamic Programming

DEFF Research Database (Denmark)

Christensen, Tue; Andersen, Kim Allan; Klose, Andreas

2013-01-01

This paper considers a minimum-cost network flow problem in a bipartite graph with a single sink. The transportation costs exhibit a staircase cost structure because such types of transportation cost functions are often found in practice. We present a dynamic programming algorithm for solving...... this so-called single-sink, fixed-charge, multiple-choice transportation problem exactly. The method exploits heuristics and lower bounds to peg binary variables, improve bounds on flow variables, and reduce the state-space variable. In this way, the dynamic programming method is able to solve large...... instances with up to 10,000 nodes and 10 different transportation modes in a few seconds, much less time than required by a widely used mixed-integer programming solver and other methods proposed in the literature for this problem....

Charge Mediated Compaction and Rearrangement of Gas-Phase Proteins: A Case Study Considering Two Proteins at Opposing Ends of the Structure-Disorder Continuum

Science.gov (United States)

Jhingree, Jacquelyn R.; Bellina, Bruno; Pacholarz, Kamila J.; Barran, Perdita E.

2017-07-01

Charge reduction in the gas phase provides a direct means of manipulating protein charge state, and when coupled to ion mobility mass spectrometry (IM-MS), it is possible to monitor the effect of charge on protein conformation in the absence of solution. Use of the electron transfer reagent 1,3-dicyanobenzene, coupled with IM-MS, allows us to monitor the effect of charge reduction on the conformation of two proteins deliberately chosen from opposite sides of the order to disorder continuum: bovine pancreatic trypsin inhibitor (BPTI) and beta casein. The ordered BPTI presents compact conformers for each of three charge states accompanied by narrow collision cross-section distributions (TWCCSDN2→He). Upon reduction of BPTI, irrespective of precursor charge state, the TWCCSN2→He decreases to a similar distribution as found for the nESI generated ion of identical charge. The behavior of beta casein upon charge reduction is more complex. It presents over a wide charge state range (9-28), and intermediate charge states (13-18) have broad TWCCSDN2→He with multiple conformations, where both compaction and rearrangement are seen. Further, we see that the TWCCSDN2→He of the latter charge states are even affected by the presence of radical anions. Overall, we conclude that the flexible nature of some proteins result in broad conformational distributions comprised of many families, even for single charge states, and the barrier between different states can be easily overcome by an alteration of the net charge.

Influence of space charge during the injection in Saturne II

International Nuclear Information System (INIS)

Lemaire, J.L.

Calculations were made in which a fixed acceptance window, established since the beginning of the injection, was considered, and a filling pattern that was a linear function of time was supposed. Evidently, these conditions are no longer met if the space charge modifies the wave number of the beam and, consequently, the filling rate changes with time. The importance of these conditions upon injection for the theoretical operation point zone 3 is reported

Highly Efficient Intracellular Protein Delivery by Cationic Polyethyleneimine-Modified Gelatin Nanoparticles

Directory of Open Access Journals (Sweden)

Ming-Ju Chou

2018-02-01

Full Text Available Intracellular protein delivery may provide a safe and non-genome integrated strategy for targeting abnormal or specific cells for applications in cell reprogramming therapy. Thus, highly efficient intracellular functional protein delivery would be beneficial for protein drug discovery. In this study, we generated a cationic polyethyleneimine (PEI-modified gelatin nanoparticle and evaluated its intracellular protein delivery ability in vitro and in vivo. The experimental results showed that the PEI-modified gelatin nanoparticle had a zeta potential of approximately +60 mV and the particle size was approximately 135 nm. The particle was stable at different biological pH values and temperatures and high protein loading efficiency was observed. The fluorescent image results revealed that large numbers of particles were taken up into the mammalian cells and escaped from the endosomes into the cytoplasm. In a mouse C26 cell-xenograft cancer model, particles accumulated in cancer cells. In conclusion, the PEI-modified gelatin particle may provide a biodegradable and highly efficient protein delivery system for use in regenerative medicine and cancer therapy.

Therapeutic intervention based on protein prenylation and associated modifications

NARCIS (Netherlands)

Gelb, M.H.; Brunsveld, L.; Hrycyna, C.A.; Michaelis, S.; Tamanoi, F.

2006-01-01

In eukaryotic cells, a specific set of proteins are modified by C-terminal attachment of 15-carbon farnesyl groups or 20-carbon geranylgeranyl groups that function both as anchors for fixing proteins to membranes and as molecular handles for facilitating binding of these lipidated proteins to other

A modified Poisson-Boltmann model including charge regulation for the adsorption of ionizable polyelectrolytes to charged interfaces, applied to lysozyme adsorption on silica

NARCIS (Netherlands)

Biesheuvel, P.M.; Veen, van der M.; Norde, W.

2005-01-01

The equilibrium adsorption of polyelectrolytes with multiple types of ionizable groups is described using a modified Poisson-Boltzmann equation including charge regulation of both the polymer and the interface. A one-dimensional mean-field model is used in which the electrostatic potential is

Origin of Asymmetric Charge Partitioning in the Dissociation of Gas-Phase Protein Homodimers

OpenAIRE

Jurchen, John C.; Williams, Evan R.

2003-01-01

The origin of asymmetric charge and mass partitioning observed for gas-phase dissociation of multiply charged macromolecular complexes has been hotly debated. These experiments hold the potential to provide detailed information about the interactions between the macromolecules within the complex. Here, this unusual phenomenon of asymmetric charge partitioning is investigated for several protein homodimers. Asymmetric charge partitioning in these ions depends on a number of factors, including ...

Characterization of Protein and Peptide Binding to Nanogels Formed by Differently Charged Chitosan Derivatives

Directory of Open Access Journals (Sweden)

Anastasia Zubareva

2013-07-01

Full Text Available Chitosan (Chi is a natural biodegradable cationic polymer with remarkable potency as a vehicle for drug or vaccine delivery. Chi possesses multiple groups, which can be used both for Chi derivatization and for particle formation. The aim of this work was to produce stable nanosized range Chi gels (nanogels, NGs with different charge and to study the driving forces of complex formation between Chi NGs and proteins or peptides. Positively charged NGs of 150 nm in diameter were prepared from hexanoyl chitosan (HC by the ionotropic gelation method while negatively charged NGs of 190 nm were obtained from succinoyl Chi (SC by a Ca2+ coacervation approach. NGs were loaded with a panel of proteins or peptides with different weights and charges. We show that NGs preferentially formed complexes with oppositely charged molecules, especially peptides, as was demonstrated by gel-electrophoresis, confocal microscopy and HPLC. Complex formation was accompanied by a change in zeta-potential and decrease in size. We concluded that complex formation between Chi NGs and peptide/proteins is mediated mostly by electrostatic interactions.

Moduli, Scalar Charges, and the First Law of Black Hole Thermodynamics

International Nuclear Information System (INIS)

Gibbons, G.; Kallosh, R.; Kol, B.

1996-01-01

We show that under variation of moduli fields φ the first law of black hole thermodynamics becomes dM=κdA/8π +ΩdJ+ψdq+χdp-Σdφ, where Σ are the scalar charges. Also the ADM mass is extremized at fixed A, J, (p,q) when the moduli fields take the fixed value φ fix (p,q) which depend only on electric and magnetic charges. Thus the double-extreme black hole minimizes the mass for fixed conserved charges. We can now explain the fact that extreme black holes fix the moduli fields at the horizon φ=φ fix (p,q): φ fix is such that the scalar charges vanish: Σ(φ fix ,(p,q))=0. copyright 1996 The American Physical Society

Tandem organic light-emitting diodes with buffer-modified C60/pentacene as charge generation layer

Science.gov (United States)

Wang, Zhen; Zheng, Xin; Liu, Fei; Wang, Pei; Gan, Lin; Wang, Jing-jing

2017-09-01

Buffer-modified C60/pentacene as charge generation layer (CGL) is investigated to achieve effective performance of charge generation. Undoped green electroluminescent tandem organic light-emitting diodes (OLEDs) with multiple identical emissive units and using buffer-modified C60/pentacene organic semiconductor heterojunction (OHJ) as CGL are demonstrated to exhibit better current density and brightness, compared with conventional single-unit devices. The current density and brightness both can be significantly improved with increasing the thickness of Al. However, excessive thickness of Al seriously decreases the transmittance of films and damages the interface. As a result, the maximum current efficiency of 1.43 cd·A-1 at 30 mA·cm-2 can be achieved for tandem OLEDs with optimal thickness of Al. These results clearly demonstrate that Cs2CO3/Al is an effective buffer for C60/pentacene-based tandem OLEDs.

Do plasma proteins distinguish between liposomes of varying charge density?

KAUST Repository

Capriotti, Anna Laura; Caracciolo, Giulio; Cavaliere, Chiara; Foglia, Patrizia; Pozzi, Daniela; Samperi, Roberto; Laganà , Aldo

2012-01-01

efficient and more biocompatible liposomal formulations, the behavior of three CLs with different membrane charge densities was investigated. The proteins of the three coronas were identified by nano-liquid chromatography-tandem mass spectrometry

Selective effects of charge on G protein activation by FSH-receptor residues 551-555 and 650-653.

Science.gov (United States)

Grasso, P; Deziel, M R; Reichert, L E

1995-01-01

Two cytosolic regions of the rat testicular FSH receptor (FSHR), residues 533-555 and 645-653, have been identified as G protein-coupling domains. We localized the activity in these domains to their C-terminal sequences, residues 551-555 (KIAKR, net charge +3) and 650-653 (RKSH, net charge +3), and examined the effects of charge on G protein activation by the C-terminal peptides, using synthetic analogs containing additions, through alanine (A) linkages, of arginine (R, +), histidine (H, +) or both. RA-KIAKR (net charge +4) mimicked the effect of FSHR-(551-555) on guanine nucleotide exchange in rat testis membranes, but reduced its ability to inhibit FSH-stimulated estradiol biosynthesis in cultured rat Sertoli cells. Further increasing net charge by the addition of H (HARA-KIAKR, net charge +5) increased guanosine 5'-triphosphate (GTP) binding, but eliminated FSHR-(551-555) effects on FSH-stimulated steroidogenesis. HA-RKSH (net charge +4) significantly inhibited guanine nucleotide exchange in rat testis membranes, but stimulated basal and potentiated FSH-induced estradiol biosynthesis in cultured rat Sertoli cells. Addition of two H residues (HAHA-RKSH, net charge +5) restored GTP binding and further potentiated basal and FSH-stimulated steroidogenesis. These results suggest that positive charges in G protein-coupling domains of the FSHR play a role in modulating G protein activation and postbinding effects of FSH, such as steroidogenesis.

Computer-aided model analysis for ionic strength-dependent effective charge of protein in ion-exchange chromatography

DEFF Research Database (Denmark)

Lim, Young-il; Jørgensen, Sten Bay; Kim, In-Ho

2005-01-01

differential algebraic equation (PDAE) system, a fast and accurate numerical method (i.e., conservation element/solution element (CE/SE) method), is proposed. Sensitivity and elasticity of the model parameters (e.g., steric/shape factors, adsorption heat coefficient, effective protein charge, equilibrium...... constant, mass transfer coefficient, axial dispersion coefficient and bed voidage) are analyzed for a BSA-salt system in a low protein concentration range. Within a low concentration range of bovine serum albumin (BSA) where linear adsorption isotherms are shown, the adsorption heat coefficient, shape...... salt concentrations, it is proposed that the effective protein charge could depend upon the salt concentration (or ionic strength). The reason for this dependence may be a steric hindrance of protein binding sites combined with a salt shielding effect neutralizing the surface charges of the protein. (c...

Impact of Rate Design Alternatives on Residential Solar Customer Bills. Increased Fixed Charges, Minimum Bills and Demand-based Rates

Energy Technology Data Exchange (ETDEWEB)

Bird, Lori [National Renewable Energy Lab. (NREL), Golden, CO (United States); Davidson, Carolyn [National Renewable Energy Lab. (NREL), Golden, CO (United States); McLaren, Joyce [National Renewable Energy Lab. (NREL), Golden, CO (United States); Miller, John [National Renewable Energy Lab. (NREL), Golden, CO (United States)

2015-09-01

With rapid growth in energy efficiency and distributed generation, electric utilities are anticipating stagnant or decreasing electricity sales, particularly in the residential sector. Utilities are increasingly considering alternative rates structures that are designed to recover fixed costs from residential solar photovoltaic (PV) customers with low net electricity consumption. Proposed structures have included fixed charge increases, minimum bills, and increasingly, demand rates - for net metered customers and all customers. This study examines the electricity bill implications of various residential rate alternatives for multiple locations within the United States. For the locations analyzed, the results suggest that residential PV customers offset, on average, between 60% and 99% of their annual load. However, roughly 65% of a typical customer's electricity demand is non-coincidental with PV generation, so the typical PV customer is generally highly reliant on the grid for pooling services.

Protein-modified nanocrystalline diamond thin films for biosensor applications.

Science.gov (United States)

Härtl, Andreas; Schmich, Evelyn; Garrido, Jose A; Hernando, Jorge; Catharino, Silvia C R; Walter, Stefan; Feulner, Peter; Kromka, Alexander; Steinmüller, Doris; Stutzmann, Martin

2004-10-01

Diamond exhibits several special properties, for example good biocompatibility and a large electrochemical potential window, that make it particularly suitable for biofunctionalization and biosensing. Here we show that proteins can be attached covalently to nanocrystalline diamond thin films. Moreover, we show that, although the biomolecules are immobilized at the surface, they are still fully functional and active. Hydrogen-terminated nanocrystalline diamond films were modified by using a photochemical process to generate a surface layer of amino groups, to which proteins were covalently attached. We used green fluorescent protein to reveal the successful coupling directly. After functionalization of nanocrystalline diamond electrodes with the enzyme catalase, a direct electron transfer between the enzyme's redox centre and the diamond electrode was detected. Moreover, the modified electrode was found to be sensitive to hydrogen peroxide. Because of its dual role as a substrate for biofunctionalization and as an electrode, nanocrystalline diamond is a very promising candidate for future biosensor applications.

Structural study of the AOT reverse micellar system. Influence of attractive interactions induced by the solubilisation of native and modified proteins

International Nuclear Information System (INIS)

Cassin, Guillaume

1994-01-01

This research thesis reports the study of the influence of intra-micellar attractions on the thermodynamic behaviour of reverse micellar systems, as well as of the effects induced by the solubilisation of natives or modified proteins. The author proposes a model to explain the decrease of attractions between droplets when the volume fraction occupied by reverse micelles increases. This model which highlights the importance of depletion forces between reverse micelles, allows the building up of a theoretical relationship between the bonding parameter and the volume fraction of reverse micelles. In order to understand the appearance of an attractive term related to the solubilisation of native cytochrome-c in these systems, this protein has been chemically modified. The author highlights the role of the charge born by a micellar probe on the thermodynamic behaviour of micro-emulsions. Then, the author applies the model of dimerizing adhesive spheres to reverse micellar systems containing native cytochrome-c. He shows that theoretical predictions of this model are in agreement with obtained experimental results [fr

Characterisation of chemically-modified proteins by electrospray ionisation mass spectrometry

International Nuclear Information System (INIS)

Bennett, K.L.

1996-09-01

Electrospray mass spectrometry (ESI-MS) has been used to examine a range of intact monoclonal antibodies (MAbs), antibody fragments such as F(ab') 2 , F ab and F c , chemically-modified fragments and a range of other chemically-modified peptides and proteins as part of a broader study aimed at establishing ESI-MS as a method for the characterisation of radioimmunoconjugates (radiolabelled monoclonal antibodies). For example, the addition of up to 10 biotin molecules to the 'papain-sensitive' 50 kDa F ab fragment can be easily detected in ESI mass spectra. For intact MAbs, however, it is only possible to detect average shifts in the mass of intact antibodies following modification. Successful ESI-MS analysis of complexes formed between chelators and other small molecules conjugated to synthetic peptides, hen egg-white Iysozyme (HEL) (M r 14 306) and horse heart myoglobin (M r 16 951) has been demonstrated. ESI-MS offers considerable advantages compared with existing methods for the characterisation of chemically-conjugated proteins including speed and sensitivity of analysis and the capability for obtaining specific structural information. The conditions for ESI-MS of intact MAbs and MAb fragments have been examined in detail and it was found that 150 kDa MAbs generally required lower sample concentration and higher skimmer potentials compared with the 50 kDa F ab fragment and other lower molecular weight proteins. In addition, the m/z range over which ions from MAbs were observed was higher (m/z ∼2000-4500) than for smaller proteins. ESI-MS was also found to be useful for probing the action of the protease papain, that is used to generate MAb fragments (F(ab) '2, F ab and F c ). Further, different sensitivities to papain for different MAb preparations was demonstrated. Finally, the tandem mass spectra of a range of peptides modified by iodine and biotin were examined. In the case of biotinylated peptides, a characteristic fragment ion was identified that could

Why do card issuers charge proportional fees?

OpenAIRE

Oz Shy; Zhu Wang

2008-01-01

This paper explains why payment card companies charge consumers and merchants fees which are proportional to the transaction values instead of charging a fixed per-transaction fee. Our theory shows that, even in the absence of any cost considerations, card companies earn much higher profit when they charge proportional fees. It is also shown that competition among merchants reduces card companies' gains from using proportional fees relative to a fixed per-transaction fee. Merchants are found ...

Strong convergence with a modified iterative projection method for hierarchical fixed point problems and variational inequalities

Directory of Open Access Journals (Sweden)

Ibrahim Karahan

2016-04-01

Full Text Available Let C be a nonempty closed convex subset of a real Hilbert space H. Let {T_{n}}:C›H be a sequence of nearly nonexpansive mappings such that F:=?_{i=1}^{?}F(T_{i}?Ø. Let V:C›H be a ?-Lipschitzian mapping and F:C›H be a L-Lipschitzian and ?-strongly monotone operator. This paper deals with a modified iterative projection method for approximating a solution of the hierarchical fixed point problem. It is shown that under certain approximate assumptions on the operators and parameters, the modified iterative sequence {x_{n}} converges strongly to x^{*}?F which is also the unique solution of the following variational inequality: ?0, ?x?F. As a special case, this projection method can be used to find the minimum norm solution of above variational inequality; namely, the unique solution x^{*} to the quadratic minimization problem: x^{*}=argmin_{x?F}?x?². The results here improve and extend some recent corresponding results of other authors.

Nonlocal continuum electrostatic theory predicts surprisingly small energetic penalties for charge burial in proteins.

Science.gov (United States)

Bardhan, Jaydeep P

2011-09-14

We study the energetics of burying charges, ion pairs, and ionizable groups in a simple protein model using nonlocal continuum electrostatics. Our primary finding is that the nonlocal response leads to markedly reduced solvent screening, comparable to the use of application-specific protein dielectric constants. Employing the same parameters as used in other nonlocal studies, we find that for a sphere of radius 13.4 Å containing a single +1e charge, the nonlocal solvation free energy varies less than 18 kcal/mol as the charge moves from the surface to the center, whereas the difference in the local Poisson model is ∼35 kcal/mol. Because an ion pair (salt bridge) generates a comparatively more rapidly varying Coulomb potential, energetics for salt bridges are even more significantly reduced in the nonlocal model. By varying the central parameter in nonlocal theory, which is an effective length scale associated with correlations between solvent molecules, nonlocal-model energetics can be varied from the standard local results to essentially zero; however, the existence of the reduction in charge-burial penalties is quite robust to variations in the protein dielectric constant and the correlation length. Finally, as a simple exploratory test of the implications of nonlocal response, we calculate glutamate pK(a) shifts and find that using standard protein parameters (ε(protein) = 2-4), nonlocal results match local-model predictions with much higher dielectric constants. Nonlocality may, therefore, be one factor in resolving discrepancies between measured protein dielectric constants and the model parameters often used to match titration experiments. Nonlocal models may hold significant promise to deepen our understanding of macromolecular electrostatics without substantially increasing computational complexity. © 2011 American Institute of Physics

Direct and inverse scattering at fixed energy for massless charged Dirac fields by Kerr-Newman-de Sitter black holes

CERN Document Server

Daudé, Thierry

2017-01-01

In this paper, the authors study the direct and inverse scattering theory at fixed energy for massless charged Dirac fields evolving in the exterior region of a Kerr-Newman-de Sitter black hole. In the first part, they establish the existence and asymptotic completeness of time-dependent wave operators associated to our Dirac fields. This leads to the definition of the time-dependent scattering operator that encodes the far-field behavior (with respect to a stationary observer) in the asymptotic regions of the black hole: the event and cosmological horizons. The authors also use the miraculous property (quoting Chandrasekhar)-that the Dirac equation can be separated into radial and angular ordinary differential equations-to make the link between the time-dependent scattering operator and its stationary counterpart. This leads to a nice expression of the scattering matrix at fixed energy in terms of stationary solutions of the system of separated equations. In a second part, the authors use this expression of ...

Nanoscale observation of local bound charges of patterned protein arrays by scanning force microscopy

International Nuclear Information System (INIS)

Oh, Y J; Jo, W; Kim, S; Park, S; Kim, Y S

2008-01-01

A protein patterned surface using micro-contact printing methods has been investigated by scanning force microscopy. Electrostatic force microscopy (EFM) was utilized for imaging the topography and detecting the electrical properties such as the local bound charge distribution of the patterned proteins. It was found that the patterned IgG proteins are arranged down to 1 μm, and the 90 deg. rotation of patterned anti-IgG proteins was successfully undertaken. Through the estimation of the effective areas, it was possible to determine the local bound charges of patterned proteins which have opposite electrostatic force behaviors. Moreover, we studied the binding probability between IgG and anti-IgG in a 1 μm 2 MIMIC system by topographic and electrostatic signals for applicable label-free detections. We showed that the patterned proteins can be used for immunoassay of proteins on the functional substrate, and that they can also be used for bioelectronics device application, indicating distinct advantages with regard to accuracy and a label-free detection

Imaging the lipidome: omega-alkynyl fatty acids for detection and cellular visualization of lipid-modified proteins.

Science.gov (United States)

Hannoush, Rami N; Arenas-Ramirez, Natalia

2009-07-17

Fatty acylation or lipid modification of proteins controls their cellular activation and diverse roles in physiology. It mediates protein-protein and protein-membrane interactions and plays an important role in regulating cellular signaling pathways. Currently, there is need for visualizing lipid modifications of proteins in cells. Herein we report novel chemical probes based on omega-alkynyl fatty acids for biochemical detection and cellular imaging of lipid-modified proteins. Our study shows that omega-alkynyl fatty acids of varying chain length are metabolically incorporated onto cellular proteins. Using fluorescence imaging, we describe the subcellular distribution of lipid-modified proteins across a panel of different mammalian cell lines and during cell division. Our results demonstrate that this methodology is a useful diagnostic tool for analyzing the lipid content of cellular proteins and for studying the dynamic behavior of lipid-modified proteins in various disease or physiological states.

Using the Ubiquitin-modified Proteome to Monitor Distinct and Spatially Restricted Protein Homeostasis Dysfunction.

Science.gov (United States)

Gendron, Joshua M; Webb, Kristofor; Yang, Bing; Rising, Lisa; Zuzow, Nathan; Bennett, Eric J

2016-08-01

Protein homeostasis dysfunction has been implicated in the development and progression of aging related human pathologies. There is a need for the establishment of quantitative methods to evaluate global protein homoeostasis function. As the ubiquitin (ub) proteasome system plays a key role in regulating protein homeostasis, we applied quantitative proteomic methods to evaluate the sensitivity of site-specific ubiquitylation events as markers for protein homeostasis dysfunction. Here, we demonstrate that the ub-modified proteome can exceed the sensitivity of engineered fluorescent reporters as a marker for proteasome dysfunction and can provide unique signatures for distinct proteome challenges which is not possible with engineered reporters. We demonstrate that combining ub-proteomics with subcellular fractionation can effectively separate degradative and regulatory ubiquitylation events on distinct protein populations. Using a recently developed potent inhibitor of the critical protein homeostasis factor p97/VCP, we demonstrate that distinct insults to protein homeostasis function can elicit robust and largely unique alterations to the ub-modified proteome. Taken together, we demonstrate that proteomic approaches to monitor the ub-modified proteome can be used to evaluate global protein homeostasis and can be used to monitor distinct functional outcomes for spatially separated protein populations. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Examination and Manipulation of Protein Surface Charge in Solution with Electrospray Ionization Mass Spectrometry

Science.gov (United States)

Gross, Deborah S.; Van Ryswyk, Hal

2014-01-01

Electrospray ionization mass spectrometry (ESI-MS) is a powerful tool for examining the charge of proteins in solution. The charge can be manipulated through choice of solvent and pH. Furthermore, solution-accessible, protonated lysine side chains can be specifically tagged with 18-crown-6 ether to form noncovalent adducts. Chemical derivatization…

The Mixed Phase of Charged AdS Black Holes

Directory of Open Access Journals (Sweden)

Piyabut Burikham

2016-01-01

Full Text Available We study the mixed phase of charged AdS black hole and radiation when the total energy is fixed below the threshold to produce a stable charged black hole branch. The coexistence conditions for the charged AdS black hole and radiation are derived for the generic case when radiation particles carry charge. The phase diagram of the mixed phase is demonstrated for both fixed potential and charge ensemble. In the dual gauge picture, they correspond to the mixed phase of quark-gluon plasma (QGP and hadron gas in the fixed chemical potential and density ensemble, respectively. In the nuclei and heavy-ion collisions at intermediate energies, the mixed phase of exotic QGP and hadron gas could be produced. The mixed phase will condense and evaporate into the hadron gas as the fireball expands.

Increased protein content of chickpea (Cicer arietinum L.) inoculated with arbuscular mycorrhizal fungi and nitrogen-fixing bacteria under water deficit conditions.

Science.gov (United States)

Oliveira, Rui S; Carvalho, Patrícia; Marques, Guilhermina; Ferreira, Luís; Nunes, Mafalda; Rocha, Inês; Ma, Ying; Carvalho, Maria F; Vosátka, Miroslav; Freitas, Helena

2017-10-01

Chickpea (Cicer arietinum L.) is a widely cropped pulse and an important source of proteins for humans. In Mediterranean regions it is predicted that drought will reduce soil moisture and become a major issue in agricultural practice. Nitrogen (N)-fixing bacteria and arbuscular mycorrhizal (AM) fungi have the potential to improve plant growth and drought tolerance. The aim of the study was to assess the effects of N-fixing bacteria and AM fungi on the growth, grain yield and protein content of chickpea under water deficit. Plants inoculated with Mesorhizobium mediterraneum or Rhizophagus irregularis without water deficit and inoculated with M. mediterraneum under moderate water deficit had significant increases in biomass. Inoculation with microbial symbionts brought no benefits to chickpea under severe water deficit. However, under moderate water deficit grain crude protein was increased by 13%, 17% and 22% in plants inoculated with M. mediterraneum, R. irregularis and M. mediterraneum + R. irregularis, respectively. Inoculation with N-fixing bacteria and AM fungi has the potential to benefit agricultural production of chickpea under water deficit conditions and to contribute to increased grain protein content. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Air stable n-doping of WSe2 by silicon nitride thin films with tunable fixed charge density

International Nuclear Information System (INIS)

Chen, Kevin; Kiriya, Daisuke; Hettick, Mark; Tosun, Mahmut; Ha, Tae-Jun; Madhvapathy, Surabhi Rao; Desai, Sujay; Sachid, Angada; Javey, Ali

2014-01-01

Stable n-doping of WSe 2 using thin films of SiN x deposited on the surface via plasma-enhanced chemical vapor deposition is presented. Positive fixed charge centers inside SiN x act to dope WSe 2 thin flakes n-type via field-induced effect. The electron concentration in WSe 2 can be well controlled up to the degenerate limit by simply adjusting the stoichiometry of the SiN x through deposition process parameters. For the high doping limit, the Schottky barrier width at the metal/WSe 2 junction is significantly thinned, allowing for efficient electron injection via tunneling. Using this doping scheme, we demonstrate air-stable WSe 2 n-MOSFETs with a mobility of ∼70 cm 2 /V s

Assessment of fixed charge density in regenerated cartilage by Gd-DTPA-enhanced MRI

International Nuclear Information System (INIS)

Miyata, Shogo; Homma, Kazuhiro; Numano, Tomokazu; Furukawa, Katsuko; Tateishi, Tetsuya; Ushida, Takashi

2006-01-01

Applying regenerated cartilage in a clinical setting requires noninvasive evaluation to detect the maturity of cartilage tissue. Magnetic resonance (MR) imaging of articular cartilage is well accepted and has been applied clinically in recent years. We attempt to establish a noninvasive method to evaluate the maturity of regenerated cartilage tissue using gadolinium-enhanced MR imaging. To reconstruct cartilaginous tissue, we embedded articular chondrocytes harvested from bovine humeral head in agarose gel and cultured the cells in vitro up to 4 weeks. The fixed charge density (FCD) of the cartilage was determined using MRI gadolinium exclusion method. The sulfated glycosaminoglycan (sGAG) content was determined by dimethylmethylene blue dye-binding assay. The sGAG content and FCD of the regenerated cartilage increased with duration of culture. In the T 1 Gd maps, the [Gd-DTPA 2- ] in the specimen decreased, and the boundary between the sample disk and the bath solution of phosphate buffered saline (PBS) became clearer as time in culture increased. In the linear regression analysis, FCD and sGAG content correlated significantly. Gadolinium-enhanced MR imaging measurements can be useful predictors of the degree of cartilaginous tissue formation. (author)

Isoelectric Point, Electric Charge, and Nomenclature of the Acid-Base Residues of Proteins

Science.gov (United States)

Maldonado, Andres A.; Ribeiro, Joao M.; Sillero, Antonio

2010-01-01

The main object of this work is to present the pedagogical usefulness of the theoretical methods, developed in this laboratory, for the determination of the isoelectric point (pI) and the net electric charge of proteins together with some comments on the naming of the acid-base residues of proteins. (Contains 8 figures and 4 tables.)

Conformal Dimensions via Large Charge Expansion.

Science.gov (United States)

Banerjee, Debasish; Chandrasekharan, Shailesh; Orlando, Domenico

2018-02-09

We construct an efficient Monte Carlo algorithm that overcomes the severe signal-to-noise ratio problems and helps us to accurately compute the conformal dimensions of large-Q fields at the Wilson-Fisher fixed point in the O(2) universality class. Using it, we verify a recent proposal that conformal dimensions of strongly coupled conformal field theories with a global U(1) charge can be obtained via a series expansion in the inverse charge 1/Q. We find that the conformal dimensions of the lowest operator with a fixed charge Q are almost entirely determined by the first few terms in the series.

Determination of the quadrupole moment of the fission-isomeric state in 238U by a modified charge-plunger method

International Nuclear Information System (INIS)

Pedersen, J.; Goerlach, U.; Habs, D.; Just, M.; Metag, V.; Mosler, E.; Schukraft, J.; Singer, P.; Specht, H.J.; Ulfert, G.

1978-01-01

A modified version of the charge plunger method has been developed which, by utilizing electrostatic fields, renders possible the separation of low- and high-charge recoil ions over short flight distances. With this technique the quadrupole moment of the 200 ns fission isomer in 238 U has been determined to be (29 +- 3) b. The corresponding deformation of c/a = (1.8 +- 0.1) provides another quantitative proof for shape isomerism in the actinide region. (orig.) [de

Influence of the Amino Acid Sequence on Protein-Mineral Interactions in Soil

Science.gov (United States)

Chacon, S. S.; Reardon, P. N.; Purvine, S.; Lipton, M. S.; Washton, N.; Kleber, M.

2017-12-01

The intimate associations between protein and mineral surfaces have profound impacts on nutrient cycling in soil. Proteins are an important source of organic C and N, and a subset of proteins, extracellular enzymes (EE), can catalyze the depolymerization of soil organic matter (SOM). Our goal was to determine how variation in the amino acid sequence could influence a protein's susceptibility to become chemically altered by mineral surfaces to infer the fate of adsorbed EE function in soil. We hypothesized that (1) addition of charged amino acids would enhance the adsorption onto oppositely charged mineral surfaces (2) addition of aromatic amino acids would increase adsorption onto zero charged surfaces (3) Increase adsorption of modified proteins would enhance their susceptibility to alterations by redox active minerals. To test these hypotheses, we generated three engineered proxies of a model protein Gb1 (IEP 4.0, 6.2 kDA) by inserting either negatively charged, positively charged or aromatic amino acids in the second loop. These modified proteins were allowed to interact with functionally different mineral surfaces (goethite, montmorillonite, kaolinite and birnessite) at pH 5 and 7. We used LC-MS/MS and solution-state Heteronuclear Single Quantum Coherence Spectroscopy NMR to observe modifications on engineered proteins as a consequence to mineral interactions. Preliminary results indicate that addition of any amino acids to a protein increase its susceptibility to fragmentation and oxidation by redox active mineral surfaces, and alter adsorption to the other mineral surfaces. This suggest that not all mineral surfaces in soil may act as sorbents for EEs and chemical modification of their structure should also be considered as an explanation for decrease in EE activity. Fragmentation of proteins by minerals can bypass the need to produce proteases, but microbial acquisition of other nutrients that require enzymes such as cellulases, ligninases or phosphatases

Orientation-dependent backbone-only residue pair scoring functions for fixed backbone protein design

Directory of Open Access Journals (Sweden)

Bordner Andrew J

2010-04-01

Full Text Available Abstract Background Empirical scoring functions have proven useful in protein structure modeling. Most such scoring functions depend on protein side chain conformations. However, backbone-only scoring functions do not require computationally intensive structure optimization and so are well suited to protein design, which requires fast score evaluation. Furthermore, scoring functions that account for the distinctive relative position and orientation preferences of residue pairs are expected to be more accurate than those that depend only on the separation distance. Results Residue pair scoring functions for fixed backbone protein design were derived using only backbone geometry. Unlike previous studies that used spherical harmonics to fit 2D angular distributions, Gaussian Mixture Models were used to fit the full 3D (position only and 6D (position and orientation distributions of residue pairs. The performance of the 1D (residue separation only, 3D, and 6D scoring functions were compared by their ability to identify correct threading solutions for a non-redundant benchmark set of protein backbone structures. The threading accuracy was found to steadily increase with increasing dimension, with the 6D scoring function achieving the highest accuracy. Furthermore, the 3D and 6D scoring functions were shown to outperform side chain-dependent empirical potentials from three other studies. Next, two computational methods that take advantage of the speed and pairwise form of these new backbone-only scoring functions were investigated. The first is a procedure that exploits available sequence data by averaging scores over threading solutions for homologs. This was evaluated by applying it to the challenging problem of identifying interacting transmembrane alpha-helices and found to further improve prediction accuracy. The second is a protein design method for determining the optimal sequence for a backbone structure by applying Belief Propagation

Study of protein-probe complexation equilibria and protein-surfactant interaction using charge transfer fluorescence probe methyl ester of N,N-dimethylamino naphthyl acrylic acid

Energy Technology Data Exchange (ETDEWEB)

Mahanta, Subrata; Balia Singh, Rupashree; Bagchi, Arnab [Department of Chemistry University of Calcutta 92, A.P.C. Road, Kolkata 700009 (India); Nath, Debnarayan [Department of Physical Chemistry, Indian Association for the Cultivation of Science, Jadavpur, Kolkata 700 032 (India); Guchhait, Nikhil, E-mail: nguchhait@yahoo.co [Department of Chemistry University of Calcutta 92, A.P.C. Road, Kolkata 700009 (India)

2010-06-15

In this paper, we demonstrate the interaction between intramolecular charge transfer (ICT) probe-Methyl ester of N,N-dimethylamino naphthyl acrylic acid (MDMANA) with bovine serum albumin (BSA) using absorption and fluorescence emission spectroscopy. The nature of probe protein binding interaction, fluorescence resonance energy transfer from protein to probe and time resolved fluorescence decay measurement predict that the probe molecule binds strongly to the hydrophobic cavity of the protein. Furthermore, the interaction of the anionic surfactant sodium dodecyl sulphate (SDS) with water soluble protein BSA has been investigated using MDMANA as fluorescenece probe. The changes in the spectral characteristics of charge transfer fluorescence probe MDMANA in BSA-SDS environment reflects well the nature of the protein-surfactant binding interaction such as specific binding, non-cooperative binding, cooperative binding and saturation binding.

Angular momentum of an electric charge and magnetically charged black hole

Energy Technology Data Exchange (ETDEWEB)

Garfinkle, D. (California Univ., Santa Barbara (USA). Dept. of Physics); Rey, S.J. (California Univ., Santa Barbara (USA). Dept. of Physics Florida Univ., Gainesville, FL (USA). Inst. for Fundamental Theory)

1991-03-21

We find the angular momentum L of a point particle with electric charge e held at a fixed position in the presence of a black hole with magnetic charge g. (For a point charge in the presence of an ordinary magnetic monopole, it is known that L=eg.) The angular momentum does depend on the separation distance between the particle and the black hole; however, L->eg for a large separation. Implications for the cosmic censorship hypothesis, the quantum hairs and other physical situations are discussed. (orig.).

Competitive Protein Adsorption on Polysaccharide and Hyaluronate Modified Surfaces

Science.gov (United States)

Ombelli, Michela; Costello, Lauren; Postle, Corinne; Anantharaman, Vinod; Meng, Qing Cheng; Composto, Russell J.; Eckmann, David M.

2011-01-01

We measured adsorption of bovine serum albumin (BSA) and fibrinogen (Fg) onto six distinct bare and dextran- and hyaluronate-modified silicon surfaces created using two dextran grafting densities and three hyaluronic acid (HA) sodium salts derived from human umbilical cord, rooster comb and streptococcus zooepidemicus. Film thickness and surface morphology depended on HA molecular weight and concentration. BSA coverage was enhanced on surfaces upon competitive adsorption of BSA:Fg mixtures. Dextranization differentially reduced protein adsorption onto surfaces based on oxidation state. Hyaluronization was demonstrated to provide the greatest resistance to protein coverage, equivalent to that of the most resistant dextranized surface. Resistance to protein adsorption was independent of the type of hyaluronic acid utilized. With changing bulk protein concentration from 20 to 40 µg ml−1 for each species, Fg coverage on silicon increased by 4×, whereas both BSA and Fg adsorption on dextran and HA were far less dependent of protein bulk concentration. PMID:21623481

Optimized localization of bacterial infections with technetium-99m labelled human immunoglobulin after protein charge selection

International Nuclear Information System (INIS)

Welling, M.; Feitsma, H.I.J.; Calame, W.; Ensing, G.J.; Goedemans, W.; Pauwels, E.K.J.

1994-01-01

To improve the scintigraphic detection of bacterial infections a protein charge-purified fraction of polyclonal human immunoglobulin was applied as a radiopharmaceutical. This purification was achieved by attaching the immunoglobulin to an anion-exchanger column and by obtaining the column-bound fraction with buffer. The binding to bacteria in vitro and the target to non-target ratios of an experimental thigh infection with Staphylococcus aureus or Klebsiella pneumoniae in mice were evaluated to compare the purified and the unpurified immunoglobulin. The percentage of binding to all gram-positive and gram-negative bacteria used in this study was significantly (P 99m Tc-labelled protein charge-purified polyclonal human immunoglobulin was administered intravenously. At all time intervals the target (infected thighs) to non-target (non-infected thighs) ratios for both infections were significantly higher (P 99m Tc-labelled protein charge-purified immunoglobulin localizes both a gram-positive and a gram-negative thigh infection more intensely and faster than 99m Tc-labelled unpurified immunoglobulin. (orig.)

Surface-treated carbon electrodes with modified potential of zero charge for capacitive deionization.

Science.gov (United States)

Wu, Tingting; Wang, Gang; Zhan, Fei; Dong, Qiang; Ren, Qidi; Wang, Jianren; Qiu, Jieshan

2016-04-15

The potential of zero charge (Epzc) of electrodes can greatly influence the salt removal capacity, charge efficiency and cyclic stability of capacitive deionization (CDI). Thus optimizing the Epzc of CDI electrodes is of great importance. A simple strategy to negatively shift the Epzc of CDI electrodes by modifying commercial activated carbon with quaternized poly (4-vinylpyridine) (AC-QPVP) is reported in this work. The Epzc of the prepared AC-QPVP composite electrode is as negative as -0.745 V vs. Ag/AgCl. Benefiting from the optimized Epzc of electrodes, the asymmetric CDI cell which consists of the AC-QPVP electrode and a nitric acid treated activated carbon (AC-HNO3) electrode exhibits excellent CDI performance. For inverted CDI, the working potential window of the asymmetric CDI cell can reach 1.4 V, and its salt removal capacity can be as high as 9.6 mg/g. For extended voltage CDI, the salt removal capacity of the asymmetric CDI cell at 1.2/-1.2 V is 20.6 mg/g, which is comparable to that of membrane CDI using pristine activated carbon as the electrodes (19.5 mg/g). The present work provides a simple method to prepare highly positively charged CDI electrodes and may pave the way for the development of high-performance CDI cells. Copyright © 2016 Elsevier Ltd. All rights reserved.

Pore structure and function of synthetic nanopores with fixed charges: tip shape and rectification properties

Energy Technology Data Exchange (ETDEWEB)

RamIrez, Patricio [Departament de Fisica Aplicada, Universitat Politecnica de Valencia, E-46022 Valencia (Spain); Apel, Pavel Yu [Flerov Laboratory of Nuclear Reactions, Joint Institute for Nuclear Research, Joliot-Curie street 6, 141980 Dubna (Russian Federation); Cervera, Javier; Mafe, Salvador [Departament de Fisica de la Terra i Termodinamica, Universitat de Valencia, E-46100 Burjassot (Spain)], E-mail: patraho@fis.upv.es

2008-08-06

We present a complete theoretical study of the relationship between the structure (tip shape and dimensions) and function (selectivity and rectification) of asymmetric nanopores on the basis of previous experimental studies. The theoretical model uses a continuum approach based on the Nernst-Planck equations. According to our results, the nanopore transport properties, such as current-voltage (I-V) characteristics, conductance, rectification ratio, and selectivity, are dictated mainly by the shape of the pore tip (we have distinguished bullet-like, conical, trumpet-like, and hybrid shapes) and the concentration of pore surface charges. As a consequence, the nanopore performance in practical applications will depend not only on the base and tip openings but also on the pore shape. In particular, we show that the pore opening dimensions estimated from the pore conductance can be very different, depending on the pore shape assumed. The results obtained can also be of practical relevance for the design of nanopores, nanopipettes, and nanoelectrodes, where the electrical interactions between the charges attached to the nanostructure and the mobile charges confined in the reduced volume of the inside solution dictate the device performance in practical applications. Because single tracks are the elementary building blocks for nanoporous membranes, the understanding and control of their individual properties should also be crucial in protein separation, water desalination, and bio-molecule detection using arrays of identical nanopores.

Pore structure and function of synthetic nanopores with fixed charges: tip shape and rectification properties

International Nuclear Information System (INIS)

RamIrez, Patricio; Apel, Pavel Yu; Cervera, Javier; Mafe, Salvador

2008-01-01

We present a complete theoretical study of the relationship between the structure (tip shape and dimensions) and function (selectivity and rectification) of asymmetric nanopores on the basis of previous experimental studies. The theoretical model uses a continuum approach based on the Nernst-Planck equations. According to our results, the nanopore transport properties, such as current-voltage (I-V) characteristics, conductance, rectification ratio, and selectivity, are dictated mainly by the shape of the pore tip (we have distinguished bullet-like, conical, trumpet-like, and hybrid shapes) and the concentration of pore surface charges. As a consequence, the nanopore performance in practical applications will depend not only on the base and tip openings but also on the pore shape. In particular, we show that the pore opening dimensions estimated from the pore conductance can be very different, depending on the pore shape assumed. The results obtained can also be of practical relevance for the design of nanopores, nanopipettes, and nanoelectrodes, where the electrical interactions between the charges attached to the nanostructure and the mobile charges confined in the reduced volume of the inside solution dictate the device performance in practical applications. Because single tracks are the elementary building blocks for nanoporous membranes, the understanding and control of their individual properties should also be crucial in protein separation, water desalination, and bio-molecule detection using arrays of identical nanopores

Pore structure and function of synthetic nanopores with fixed charges: tip shape and rectification properties.

Science.gov (United States)

Ramírez, Patricio; Apel, Pavel Yu; Cervera, Javier; Mafé, Salvador

2008-08-06

We present a complete theoretical study of the relationship between the structure (tip shape and dimensions) and function (selectivity and rectification) of asymmetric nanopores on the basis of previous experimental studies. The theoretical model uses a continuum approach based on the Nernst-Planck equations. According to our results, the nanopore transport properties, such as current-voltage (I-V) characteristics, conductance, rectification ratio, and selectivity, are dictated mainly by the shape of the pore tip (we have distinguished bullet-like, conical, trumpet-like, and hybrid shapes) and the concentration of pore surface charges. As a consequence, the nanopore performance in practical applications will depend not only on the base and tip openings but also on the pore shape. In particular, we show that the pore opening dimensions estimated from the pore conductance can be very different, depending on the pore shape assumed. The results obtained can also be of practical relevance for the design of nanopores, nanopipettes, and nanoelectrodes, where the electrical interactions between the charges attached to the nanostructure and the mobile charges confined in the reduced volume of the inside solution dictate the device performance in practical applications. Because single tracks are the elementary building blocks for nanoporous membranes, the understanding and control of their individual properties should also be crucial in protein separation, water desalination, and bio-molecule detection using arrays of identical nanopores.

Multilayer Choline Phosphate Molecule Modified Surface with Enhanced Cell Adhesion but Resistance to Protein Adsorption.

Science.gov (United States)

Chen, Xingyu; Yang, Ming; Liu, Botao; Li, Zhiqiang; Tan, Hong; Li, Jianshu

2017-08-22

Choline phosphate (CP), which is a new zwitterionic molecule, and has the reverse order of phosphate choline (PC) and could bind to the cell membrane though the unique CP-PC interaction. Here we modified a glass surface with multilayer CP molecules using surface-initiated atom-transfer radical polymerization (SI-ATRP) and the ring-opening method. Polymeric brushes of (dimethylamino)ethyl methacrylate (DMAEMA) were synthesized by SI-ATRP from the glass surface. Then the grafted PDMAEMA brushes were used to introduce CP groups to fabricate the multilayer CP molecule modified surface. The protein adsorption experiment and cell culture test were used to evaluate the biocompatibility of the modified surfaces by using human umbilical veinendothelial cells (HUVECs). The protein adsorption results demonstrated that the multilayer CP molecule decorated surface could prevent the adsorption of fibrinogen and serum protein. The adhesion and proliferation of cells were improved significantly on the multilayer CP molecule modified surface. Therefore, the biocompatibility of the material surface could be improved by the modified multilayer CP molecule, which exhibits great potential for biomedical applications, e.g., scaffolds in tissue engineering.

Angular momentum of an electric charge and magnetically charged black hole

International Nuclear Information System (INIS)

Garfinkle, D.; Rey, Soo-Jong

1990-01-01

We find the angular momentum L of a point particle with electric charge e held at a fixed position in the presence of a black hole with magnetic charge g. (For a point charge in the presence of an of ordinary magnetic monopole, it is known that L = eg). The angular momentum does depend on the separation distance between the particle and the black hole; however, L → eg for a large separation. Implications for the cosmic censorship hypothesis, the quantum hairs and other physical situations are discussed

Effect of alterations in glomerular charge on deposition of cationic and anionic antibodies to fixed glomerular antigens in the rat.

Science.gov (United States)

Adler, S; Baker, P; Pritzl, P; Couser, W G

1985-07-01

Reduction of the negative charge of the glomerular capillary wall alters its charge- and size-selective properties. To investigate the effect of alteration in glomerular charge properties on antibody localization, we prepared cationic and anionic fractions of antibodies to subepithelial and glomerular basement membrane (GBM) antigens, and compared their deposition in normal rats and rats treated with protamine sulfate or aminonucleoside of puromycin to reduce capillary wall charge. IgG antibodies were eluted from kidneys of rats with active Heymann's nephritis (AICN), passive Heymann's nephritis (PHN), or anti-GBM nephritis (NTN), separated into cationic and anionic fractions, and radiolabeled with iodine 125 or iodine 131. Relative antibody content of each fraction was determined by incubation with an excess of glomerular antigen. Varying amounts of cationic and anionic IgG eluted from kidneys of rats with AICN or PHN were injected into 24 normal or protamine sulfate-treated rats. Glomerular binding of all antibodies was highly correlated with IgG delivery to the kidney. The ratio of cationic to anionic antibody deposited in the glomeruli of normal rats after 4 hours was 1.08 +/- 0.07 for AICN eluate and 0.37 +/- 0.04 for PHN eluate. The ratios were not significantly different in animals pretreated with protamine sulfate (1.15 +/- 0.06 and 0.44 +/- 0.06, respectively; P greater than 0.05). Varying amounts of cationic and anionic IgG eluted from kidneys of rats with NTN were injected into 10 normal rats and four rats treated with aminonucleoside of puromycin. Glomerular binding of antibody was again highly correlated with IgG delivery to the kidney. The ratio of cationic to anionic antibody deposited in the glomeruli of normal rats after 1 hour was 1.03 +/- 0.06, and was not significantly altered in rats treated with aminonucleoside of puromycin (1.05 +/- 0.03, P greater than 0.5). Proteinuria in PHN rats was also unaffected by treatment with protamine sulfate for

Modulating Protein Adsorption on Oxygen Plasma Modified Polysiloxane Surfaces

International Nuclear Information System (INIS)

Marletta, G.

2006-01-01

In the present paper we report the study on the adsorption behaviour of three model globular proteins, Human Serum Albumin, Lactoferrin and Egg Chicken Lysozyme onto both unmodified surfaces of a silicon-based polymer and the corresponding plasma treated surfaces. In particular, thin films of hydrophobic polysiloxane (about 90 degree of static water contact angle, WCA) were converted by oxygen plasma treatment at reduced pressure into very hydrophilic phases of SiOx (WCA less than 5 degree). The kinetics of protein adsorption processes were investigated by QCM-D technique, while the chemical structure and topography of the protein adlayer have been studied by Angular resolved-XPS and AFM respectively. It turned out that Albumin and Lysozyme exhibited the opposite preferential adsorption respectively onto the hydrophobic and hydrophilic surfaces, while Lactoferrin did not exhibit significant differences. The observed protein behaviour are discussed both in terms of surface-dependent parameters, including surface free energy and chemical structure, and in terms of protein-dependent parameters, including charge as well as the average molecular orientation in the adlayers. Finally, some examples of differential adsorption behaviour of the investigated proteins are reported onto nanopatterned polysiloxane surfaces consisting of hydrophobic nanopores surrounded by hydrophilic (plasma-treated) matrix and the reverse

Modified Pattern Sequence-based Forecasting for Electric Vehicle Charging Stations

Energy Technology Data Exchange (ETDEWEB)

Majidpour, Mostafa; Qiu, Charlie; Chu, Peter; Gadh, Rajit; Pota, Hemanshu R.

2014-11-03

Three algorithms for the forecasting of energy consumption at individual EV charging outlets have been applied to real world data from the UCLA campus. Out of these three algorithms, namely k-Nearest Neighbor (kNN), ARIMA, and Pattern Sequence Forecasting (PSF), kNN with k=1, was the best and PSF was the worst performing algorithm with respect to the SMAPE measure. The advantage of PSF is its increased robustness to noise by substituting the real valued time series with an integer valued one, and the advantage of NN is having the least SMAPE for our data. We propose a Modified PSF algorithm (MPSF) which is a combination of PSF and NN; it could be interpreted as NN on integer valued data or as PSF with considering only the most recent neighbor to produce the output. Some other shortcomings of PSF are also addressed in the MPSF. Results show that MPSF has improved the forecast performance.

76 FR 10811 - Annual Charges for Use of Government Lands

Science.gov (United States)

2011-02-28

... in fixing such charges the Commission shall seek to avoid increasing the price to the consumers of... accurately reflected current economic conditions and conformed precisely to the context of land used for... to Consumers of Power 24. In fixing annual charges, we must seek to avoid increasing the price to...

Testing effective string models of black holes with fixed scalars

International Nuclear Information System (INIS)

Krasnitz, M.; Klebanov, I.R.

1997-01-01

We solve the problem of mixing between the fixed scalar and metric fluctuations. First, we derive the decoupled fixed scalar equation for the four-dimensional black hole with two different charges. We proceed to the five-dimensional black hole with different electric (one-brane) and magnetic (five-brane) charges, and derive two decoupled equations satisfied by appropriate mixtures of the original fixed scalar fields. The resulting greybody factors are proportional to those that follow from coupling to dimension (2,2) operators on the effective string. In general, however, the string action also contains couplings to chiral operators of dimension (1,3) and (3,1), which cause disagreements with the semiclassical absorption cross sections. Implications of this for the effective string models are discussed. copyright 1997 The American Physical Society

Field effect of screened charges: electrical detection of peptides and proteins by a thin-film resistor.

Science.gov (United States)

Lud, Simon Q; Nikolaides, Michael G; Haase, Ilka; Fischer, Markus; Bausch, Andreas R

2006-02-13

For many biotechnological applications the label-free detection of biomolecular interactions is becoming of outstanding importance. In this Article we report the direct electrical detection of small peptides and proteins by their intrinsic charges using a biofunctionalized thin-film resistor. The label-free selective and quantitative detection of small peptides and proteins is achieved using hydrophobized silicon-on-insulator (SOI) substrates functionalized with lipid membranes that incorporate metal-chelating lipids. The response of the nanometer-thin conducting silicon film to electrolyte screening effects is taken into account to determine quantitatively the charges of peptides. It is even possible to detect peptides with a single charge and to distinguish single charge variations of the analytes even in physiological electrolyte solutions. As the device is based on standard semiconductor technologies, parallelization and miniaturization of the SOI-based biosensor is achievable by standard CMOS technologies and thus a promising basis for high-throughput screening or biotechnological applications.

Comparison of Zirconium Phosphonate-Modified Surfaces for Immobilizing Phosphopeptides and Phosphate-Tagged Proteins.

Science.gov (United States)

Forato, Florian; Liu, Hao; Benoit, Roland; Fayon, Franck; Charlier, Cathy; Fateh, Amina; Defontaine, Alain; Tellier, Charles; Talham, Daniel R; Queffélec, Clémence; Bujoli, Bruno

2016-06-07

Different routes for preparing zirconium phosphonate-modified surfaces for immobilizing biomolecular probes are compared. Two chemical-modification approaches were explored to form self-assembled monolayers on commercially available primary amine-functionalized slides, and the resulting surfaces were compared to well-characterized zirconium phosphonate monolayer-modified supports prepared using Langmuir-Blodgett methods. When using POCl3 as the amine phosphorylating agent followed by treatment with zirconyl chloride, the result was not a zirconium-phosphonate monolayer, as commonly assumed in the literature, but rather the process gives adsorbed zirconium oxide/hydroxide species and to a lower extent adsorbed zirconium phosphate and/or phosphonate. Reactions giving rise to these products were modeled in homogeneous-phase studies. Nevertheless, each of the three modified surfaces effectively immobilized phosphopeptides and phosphopeptide tags fused to an affinity protein. Unexpectedly, the zirconium oxide/hydroxide modified surface, formed by treating the amine-coated slides with POCl3/Zr(4+), afforded better immobilization of the peptides and proteins and efficient capture of their targets.

DNA-modified electrodes fabricated using copper-free click chemistry for enhanced protein detection.

Science.gov (United States)

Furst, Ariel L; Hill, Michael G; Barton, Jacqueline K

2013-12-31

A method of DNA monolayer formation has been developed using copper-free click chemistry that yields enhanced surface homogeneity and enables variation in the amount of DNA assembled; extremely low-density DNA monolayers, with as little as 5% of the monolayer being DNA, have been formed. These DNA-modified electrodes (DMEs) were characterized visually, with AFM, and electrochemically, and were found to facilitate DNA-mediated reduction of a distally bound redox probe. These low-density monolayers were found to be more homogeneous than traditional thiol-modified DNA monolayers, with greater helix accessibility through an increased surface area-to-volume ratio. Protein binding efficiency of the transcriptional activator TATA-binding protein (TBP) was also investigated on these surfaces and compared to that on DNA monolayers formed with standard thiol-modified DNA. Our low-density monolayers were found to be extremely sensitive to TBP binding, with a signal decrease in excess of 75% for 150 nM protein. This protein was detectable at 4 nM, on the order of its dissociation constant, with our low-density monolayers. The improved DNA helix accessibility and sensitivity of our low-density DNA monolayers to TBP binding reflects the general utility of this method of DNA monolayer formation for DNA-based electrochemical sensor development.

Heterologous protein secretion in Lactobacilli with modified pSIP vectors.

Directory of Open Access Journals (Sweden)

Ingrid Lea Karlskås

Full Text Available We describe new variants of the modular pSIP-vectors for inducible gene expression and protein secretion in lactobacilli. The basic functionality of the pSIP system was tested in Lactobacillus strains representing 14 species using pSIP411, which harbors the broad-host-range Lactococcus lactis SH71rep replicon and a β-glucuronidase encoding reporter gene. In 10 species, the inducible gene expression system was functional. Based on these results, three pSIP vectors with different signal peptides were modified by replacing their narrow-host-range L. plantarum 256rep replicon with SH71rep and transformed into strains of five different species of Lactobacillus. All recombinant strains secreted the target protein NucA, albeit with varying production levels and secretion efficiencies. The Lp_3050 derived signal peptide generally resulted in the highest levels of secreted NucA. These modified pSIP vectors are useful tools for engineering a wide variety of Lactobacillus species.

Photoinduced ultrafast charge-order melting: Charge-order inversion and nonthermal effects

International Nuclear Information System (INIS)

Veenendaal, Michel van

2016-01-01

The effect of photoexcitation is studied for a system with checkerboard charge order induced by displacements of ligands around a metal site. The motion of the ligands is treated classically and the electronic charges are simplified to two-level molecular bond charges. The calculations are done for a checkerboard charge-ordered system with about 100 000 ligand oscillators coupled to a fixed-temperature bath. The initial photoexcitation is followed by a rapid decrease in the charge-order parameter within 50–100 femtoseconds while leaving the correlation length almost unchanged. Depending on the fluence, a complete melting of the charge order occurs in less than a picosecond. While for low fluences, the system returns to its original state, for full melting, it recovers to its broken-symmetry state leading to an inversion of the charge order. Finally, for small long-range interactions, recovery can be slow due to domain formation.

Charge transfer processes on electrodes modified by polymer films of metal complexes with Schiff bases

International Nuclear Information System (INIS)

Levin, O.V.; Karushev, M.P.; Timonov, A.M.; Alekseeva, E.V.; Zhang, Shuanghua; Malev, V.V.

2013-01-01

Electrochemical properties of glassy carbon electrodes modified by two polymer films of different nickel complexes with Schiff base ligands containing methoxy substituents in their aromatic parts were studied in acetonitrile solutions with cyclic voltammetry, quartz crystal microbalance, atomic force microscopy, and impedance spectroscopy. It was observed that introduction of such substituents leads to a noticeable splitting of cycling voltammetric curves into at least two ox/red transitions. In addition, solvent flows accompanying the counter-ions ones during charging/discharge processes within the films appeared significantly greater than those observed in the case of non-substituted ligands. The obtained impedance results as a whole were satisfactorily treated in scope of the so-called model of homogeneous films with two kinds of charge carriers. However, determinations of the Warburg constant as a function of the electrode potential require additional verifications, at least in the ranges of overlapping ox/red transitions. In particular, it was established that in this region the impedance frequency dependence was some superposition of the diffusion (Warburg) and the pseudo-capacitive constituents. This, most likely, resulted from the presence of three kinds of charge carriers in the film interior

Pepsin-Assisted Transglutaminase Modifi cation of Functional Properties of a Protein Isolate Obtained from Industrial Sunfl ower Meal

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Petya Ivanova

2017-01-01

Full Text Available The utilization of industrial sunfl ower meal to produce protein-rich products for the food industry is an alternative approach for bett er and more effi cient use of this agricultural by-product. Sunfl ower meal proteins possess specifi c functional properties, which however need improvement to broaden their potential as supplements for delivering high-quality products for human nutrition. The aim of the study is to evaluate the combined infl uence of low-degree pepsin hydrolysis and transglutaminase (TG modifi cation on industrial sunfl ower meal protein isolate functionality at pH=2 to 10. Three TG-modifi ed pepsin hydrolysates with the degree of hydrolysis of 0.48, 0.71 and 1.72 % were produced and named TG-PH1, TG-PH2 and TG-PH3, respectively. All three TG-modifi ed pepsin hydrolysates exhibited improved solubility at pH between 3.5 and 5.5 as the highest was observed of TG-PH3 at protein isoelectric point (pI=4.5. Sunfl ower meal protein isolate and TG-modifi ed sunfl ower meal protein isolate had greater solubility than the three TG-modifi ed hydrolysates at pH7. Signifi cant improvement of foam making capacity (p<0.05 was achieved with all three TG-modifi ed pepsin hydrolysates in the entire pH area studied. Pepsin hydrolysis of the protein isolate with the three degrees of hydrolysis did not improve foam stability. Improved thermal stability was observed with TG-PH3 up to 80 °C compared to the protein isolate (pH=7. At 90 °C, TG modifi cation of the protein isolate alone resulted in the highest thermal stability. Pepsin hydrolysis followed by a treatment with TG could be used to produce sunfl ower protein isolates with improved solubility, foam making capacity and thermal stability for use in the food industry.

MOSFET Electric-Charge Sensor

Science.gov (United States)

Robinson, Paul A., Jr.

1988-01-01

Charged-particle probe compact and consumes little power. Proposed modification enables metal oxide/semiconductor field-effect transistor (MOSFET) to act as detector of static electric charges or energetic charged particles. Thickened gate insulation acts as control structure. During measurements metal gate allowed to "float" to potential of charge accumulated in insulation. Stack of modified MOSFET'S constitutes detector of energetic charged particles. Each gate "floats" to potential induced by charged-particle beam penetrating its layer.

Effect of reagent charge on the labeling of erythrocyte membrane proteins by photoactivated reagents

International Nuclear Information System (INIS)

Schaeffer, J.C.; Hakimian, R.; Shimer, M.L.

1986-01-01

Leaky erythrocyte ghosts were labeled with 3 H-[2-(4-azido-2-nitroanilino)ethyl]trimethylammonium iodide (cationic label) or 3 H-N-(4-azido-2-nitrophenyl)-β-alanine (anionic label). After the membranes were thoroughly washed, seven times as much cationic label was associated with the membranes as anionic label at 5 μM, whereas at 50 μM the cationic label was favored 15-fold. The distribution of label in the membrane proteins was ascertain by SDS-gel electrophoresis followed by autoradiography. At 50 μM cationic label, erythrocyte membrane protein bands 1,2,3,4.2, and 5 were intensely labeled, while band 6 was labeled weakly. At 5 μM cationic label, bands 1 and 4.2 were heavily labeled, while 2,3 and 5 were labeled less well. At both 50 μM and 5 μM anionic label, bands 1 and 6 were most prominently labeled. Bands 2,3,4.2 and 5 were labeled also at 50 μM, but they were labeled only very weakly at 5 μM. Band 4.1 was labeled very poorly if at all by either reagent. A mixture of the reagents gave an additive pattern. Thus, the charge and concentration of these reagents appear to play a major role in their ability to label membrane proteins indiscriminately. Because these reagents contain the same chromophore, 4-azido-2-nitroaniline, and differ mainly only in their charge, they may prove useful in assessing the location of charged sites on proteins in supramolecular complexes

Feasibility analysis of color removal from textile dyeing wastewater in a fixed-bed column system by surfactant-modified zeolite (SMZ)

International Nuclear Information System (INIS)

Ozdemir, Ozgur; Turan, Mustafa; Turan, Abdullah Zahid; Faki, Aysegul; Engin, Ahmet Baki

2009-01-01

In this study, the ability of surfactant-modified zeolite (SMZ) to remove color from real textile wastewater was investigated. Tests were performed in a fixed-bed column reactor and the surface of natural zeolite was modified with a quaternary amine surfactant hexadecyltrimethylammonium bromide (HTAB). The zeolite bed that was modified at 1 g L -1 HTAB concentration and HTAB flow rate of 0.015 L min -1 showed good performance in removing color. Effects of wastewater color intensity, flow rates and bed heights were also studied. Wastewater was diluted several times in the ratios of 25%, 50% and 75% in order to assess the influence of wastewater strength. The breakthrough curves of the original and diluted wastewaters are dispersed due to the fact that breakthrough came late at lower color intensities and saturation of the bed appeared faster at higher color intensities. The column had a 3-cm diameter and four different bed heights of 12.5, 25, 37.5 and 50 cm, which treated 5.25, 19.50, 35.25 and 51 L original textile wastewater, respectively, at the breakthrough time at a flow rate of 0.025 L min -1 . The theoretical service times evaluated from bed depth service time (BDST) approach for different column variables. The calculated and theoretical values of the exchange zone height were found with a difference of 27%. The various design parameters obtained from fixed-bed experimental studies showed good correlation with corresponding theoretical values, under different bed heights. The regeneration of the SMZ was also evaluated using a solution consisting of 30 g L -1 NaCl and 1.5 g L -1 NaOH at pH 12 and temperature 30 o C. Twice-regenerated SMZ showed the best performance compared with the others while first- and thrice-regenerated perform lower than the original SMZ.

A modified FASP protocol for high-throughput preparation of protein samples for mass spectrometry.

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Jeremy Potriquet

Full Text Available To facilitate high-throughput proteomic analyses we have developed a modified FASP protocol which improves the rate at which protein samples can be processed prior to mass spectrometry. Adapting the original FASP protocol to a 96-well format necessitates extended spin times for buffer exchange due to the low centrifugation speeds tolerated by these devices. However, by using 96-well plates with a more robust polyethersulfone molecular weight cutoff membrane, instead of the cellulose membranes typically used in these devices, we could use isopropanol as a wetting agent, decreasing spin times required for buffer exchange from an hour to 30 minutes. In a typical work flow used in our laboratory this equates to a reduction of 3 hours per plate, providing processing times similar to FASP for the processing of up to 96 samples per plate. To test whether our modified protocol produced similar results to FASP and other FASP-like protocols we compared the performance of our modified protocol to the original FASP and the more recently described eFASP and MStern-blot. We show that all FASP-like methods, including our modified protocol, display similar performance in terms of proteins identified and reproducibility. Our results show that our modified FASP protocol is an efficient method for the high-throughput processing of protein samples for mass spectral analysis.

Synthesis and structural characterization of carboxyethylpyrrole-modified proteins: mediators of age-related macular degeneration.

Science.gov (United States)

Lu, Liang; Gu, Xiaorong; Hong, Li; Laird, James; Jaffe, Keeve; Choi, Jaewoo; Crabb, John; Salomon, Robert G

2009-11-01

Protein modifications in which the epsilon-amino group of lysyl residues is incorporated into a 2-(omega-carboxyethyl)pyrrole (CEP) are mediators of age-related macular degeneration (AMD). They promote both angiogenesis into the retina ('wet AMD') and geographic retinal atrophy ('dry AMD'). Blood levels of CEPs are biomarkers for clinical prognosis of the disease. To enable mechanistic studies of their role in promoting AMD, for example, through the activation of B- and T-cells, interaction with receptors, or binding with complement proteins, we developed an efficient synthesis of CEP derivatives, that is especially effective for proteins. The structures of tryptic peptides derived from CEP-modified proteins were also determined. A key finding is that 4,7-dioxoheptanoic acid 9-fluorenylmethyl ester reacts with primary amines to provide 9-fluorenylmethyl esters of CEP-modified proteins that can be deprotected in situ with 1,8-diazabicyclo[5.4.0]undec-7-ene without causing protein denaturation. The introduction of multiple CEP-modifications with a wide variety of CEP:protein ratios is readily achieved using this strategy.

Isolation of pronephros cells which endocytose chemically modified proteins in the rainbow trout

International Nuclear Information System (INIS)

Dannevig, B.H.; Berg, T.

1986-01-01

Modified serum albumin is cleared from the blood by kidney cells in salmonid fishes. The present study deals with isolation of cells from pronephros which endocytose formaldehyde-treated human serum albumin (fHSA). Radioactively labelled fHSA or dinitrophenyl-conjugated albumin (DNP-HSA) were injected intravenously into rainbow trouts. Pronephros cells, containing the endocytosed protein, were isolated and further separated by centrifugal elutriation and density-gradient centrifugation. Most of the radioactive protein was elutriated together with small cells. After centrifuging the cells through a Percoll density gradient, radioactive protein was located in cells recovered in the upper part of the gradient. In mammals, fHSA and other modified proteins are mainly taken up by sinusoidal endothelial cells in the liver via a scavenger receptor 0. Our results suggest that a comparable function in salmonids is located in a subpopulation of relatively small cells in kidney tissue, possibly sinusoidal lining cells. The separation techniques used seemed to be suitable for isolation of different populations of pronephros cells

The IGF-I/IGFBP-3 system in gingival crevicular fluid and dependence on application of fixed force.

Science.gov (United States)

Toia, M; Galazzo, R; Maioli, C; Granata, R; Scarlatti, F

2005-12-01

During application of orthodontic force on the tooth, various molecular parameters associated with tissue remodeling are changed. IGF-I is a regulatory protein produced during periodontal regeneration. IGF binding proteins-3 (IGFBP-3), a specific IGF-I binding protein, is the major regulatory factor of IGF-I activity. We tested the hypothesis that changes in the IGF-I/ IGFBP-3 system occur during fixed force application to the tooth and that these changes are detectable in the gingival crevicular fluid (GCF). IGFBP-3 and IGF-I secretion into gingival crevicular fluid (GCF) was analyzed by Western blotting and immunoradiometric assay (IRMA), respectively, in GCF of 6 healthy subjects just prior to and during orthodontics treatment using fixed appliances. We observed a significant time-dependent decrease of IGFBP-3 content in GCF during orthodontic treatment (4 h and 10 days). Reduction in levels of intact, glycosylated 47 kDa form of IGFBP-3 was associated with its degradation and the appearance of intermediate breakdown products. IGF-I levels were significantly increased 4 h after application of orthodontic force, while they were significantly reduced 10 days after the start of treatment. IGFBP-3 secretion into GCF and its molecular structure are modified by the fixed force of orthodontic treatment. Alterations in IGFBP-3 appear to be unrelated to the binding to IGF-I, suggesting an IGF-independent role of this binding protein in tooth movement.

Preparation of Modified Films with Protein from Grouper Fish

Science.gov (United States)

Tecante, A.; Granados-Navarrete, S.; Martínez-García, C.

2016-01-01

A protein concentrate (PC) was obtained from Grouper fish skin and it was used to prepare films with different amounts of sorbitol and glycerol as plasticizers. The best performing films regarding resistance were then modified with various concentrations of CaCl2, CaSO4 (calcium salts), and glucono-δ-lactone (GDL) with the purpose of improving their mechanical and barrier properties. These films were characterized by determining their mechanical properties and permeability to water vapor and oxygen. Formulations with 5% (w/v) protein and 75% sorbitol and 4% (w/v) protein with a mixture of 15% glycerol and 15% sorbitol produced adequate films. Calcium salts and GDL increased the tensile fracture stress but reduced the fracture strain and decreased water vapor permeability compared with control films. The films prepared represent an attractive alternative for being used as food packaging materials. PMID:27597950

Potential allergenicity research of Cry1C protein from genetically modified rice.

Science.gov (United States)

Cao, Sishuo; He, Xiaoyun; Xu, Wentao; Luo, Yunbo; Ran, Wenjun; Liang, Lixing; Dai, Yunqing; Huang, Kunlun

2012-07-01

With the development of genetically modified crops, there has been a growing interest in available approaches to assess the potential allergenicity of novel gene products. We were not sure whether Cry1C could induce allergy. We examined the protein with three other proteins to determine the potential allergenicity of Cry1C protein from genetically modified rice. Female Brown Norway (BN) rats received 0.1 mg peanut agglutinin (PNA), 1mg potato acid phosphatase (PAP), 1mg ovalbumin (OVA) or 5 mg purified Cry1C protein dissolved in 1 mL water by daily gavage for 42 days to test potential allergenicity. Ten days after the last gavage, rats were orally challenged with antigens, and physiologic and immunologic responses were studied. In contrast to sensitization with PNA, PAP and OVA Cry1C protein did not induce antigen-specific IgG2a in BN rats. Cytokine expression, serum IgE and histamine levels and the number of eosinophils and mast cells in the blood of Cry1C group rats were comparable to the control group rats, which were treated with water alone. As Cry1C did not show any allergenicity, we make the following conclusion that the protein could be safety used in rice or other plants. Copyright © 2012 Elsevier Inc. All rights reserved.

Lectures on Antitrust Economics, Chapter 2: Price Fixing

OpenAIRE

Whinston, Michael D.

2003-01-01

In this chapter, we begin our discussion of antitrust economics by considering what many consider its most central element: its ban on "price fixing" - that is, agreements among competitors over the prices they will charge or the outputs they will produce. Indeed, the prohibition on price fixing is one area of antitrust law that even those generally skeptical of governmental compe- tition policy typically regard approvingly. Nevertheless, despite its current uncontroversial status, we shall s...

Differences between charge-charge forward-backward multiplicity correlations in multiparticle production

International Nuclear Information System (INIS)

Barshay, S.

1987-01-01

We analyze new data from hadron-hadron collisions on forward-backward correlations. The difference between these correlations for unlike and like charges implies different dynamical correlations at fixed impact parameter. These different correlations should be observable in e + e - annihilations. (orig.)

Amino acid size, charge, hydropathy indices and matrices for protein structure analysis

Directory of Open Access Journals (Sweden)

Biro JC

2006-03-01

Full Text Available Abstract Background Prediction of protein folding and specific interactions from only the sequence (ab initio is a major challenge in bioinformatics. It is believed that such prediction will prove possible if Anfinsen's thermodynamic principle is correct for all kinds of proteins, and all the information necessary to form a concrete 3D structure is indeed present in the sequence. Results We indexed the 200 possible amino acid pairs for their compatibility regarding the three major physicochemical properties – size, charge and hydrophobicity – and constructed Size, Charge and Hydropathy Compatibility Indices and Matrices (SCI & SCM, CCI & CCM, and HCI & HCM. Each index characterized the expected strength of interaction (compatibility of two amino acids by numbers from 1 (not compatible to 20 (highly compatible. We found statistically significant positive correlations between these indices and the propensity for amino acid co-locations in real protein structures (a sample containing total 34630 co-locations in 80 different protein structures: for HCI: p We tried to predict or reconstruct simple 2D representations of 3D structures from the sequence using these matrices by applying a dot plot-like method. The location and pattern of the most compatible subsequences was very similar or identical when the three fundamentally different matrices were used, which indicates the consistency of physicochemical compatibility. However, it was not sufficient to choose one preferred configuration between the many possible predicted options. Conclusion Indexing of amino acids for major physico-chemical properties is a powerful approach to understanding and assisting protein design. However, it is probably insufficient itself for complete ab initio structure prediction.

Charge reversal and surface charge amplification in asymmetric valence restricted primitive model planar electric double layers in the modified Poisson-Boltzmann theory

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L.B. Bhuiyan

2017-12-01

Full Text Available The modified Poisson-Boltzmann theory of the restricted primitive model double layer is revisited and recast in a fresh, slightly broader perspective. Derivation of relevant equations follow the techniques utilized in the earlier MPB4 and MPB5 formulations and clarifies the relationship between these. The MPB4, MPB5, and a new formulation of the theory are employed in an analysis of the structure and charge reversal phenomenon in asymmetric 2:1/1:2 valence electrolytes. Furthermore, polarization induced surface charge amplification is studied in 3:1/1:3 systems. The results are compared to the corresponding Monte Carlo simulations. The theories are seen to predict the "exact" simulation data to varying degrees of accuracy ranging from qualitative to almost quantitative. The results from a new version of the theory are found to be of comparable accuracy as the MPB5 results in many situations. However, in some cases involving low electrolyte concentrations, theoretical artifacts in the form of un-physical "shoulders" in the singlet ionic distribution functions are observed.

Feasibility analysis of As(III) removal in a continuous flow fixed bed system by modified calcined bauxite (MCB)

International Nuclear Information System (INIS)

Bhakat, P.B.; Gupta, A.K.; Ayoob, S.

2007-01-01

This study examine the feasibility of As(III) removal from aqueous environment by an adsorbent, modified calcined bauxite (MCB) in a continuous flow fixed bed system. MCB exhibited excellent adsorption capacity of 520.2 mg/L (0.39 mg/g) with an adsorption rate constant 0.7658 L/mg h for an influent As(III) concentration of 1 mg/L. In a 2 cm diameter continuous flow fixed MCB bed, a depth of only 1.765 cm was found necessary to produce effluent As(III) concentration of 0.01 mg/L, from an influent of 1 mg/L at a flow rate of 8 mL/min. Also, bed heights of 10, 20, and 30 cm could treat 427.85, 473.88 and 489.17 bed volumes of water, respectively, to breakthrough. A reduction in adsorption capacity of MCB was observed with increase in flow rates. The theoretical service times evaluated from bed depth service time (BDST) approach for different flow rates and influent As(III) concentrations had shown good correlation with the corresponding experimental values. The theoretical breakthrough curve developed from constantly mixed batch reactor (CMBR) isotherm data also correlated well with experimental breakthrough curve

Protein classification using modified n-grams and skip-grams.

Science.gov (United States)

Islam, S M Ashiqul; Heil, Benjamin J; Kearney, Christopher Michel; Baker, Erich J

2018-05-01

Classification by supervised machine learning greatly facilitates the annotation of protein characteristics from their primary sequence. However, the feature generation step in this process requires detailed knowledge of attributes used to classify the proteins. Lack of this knowledge risks the selection of irrelevant features, resulting in a faulty model. In this study, we introduce a supervised protein classification method with a novel means of automating the work-intensive feature generation step via a Natural Language Processing (NLP)-dependent model, using a modified combination of n-grams and skip-grams (m-NGSG). A meta-comparison of cross-validation accuracy with twelve training datasets from nine different published studies demonstrates a consistent increase in accuracy of m-NGSG when compared to contemporary classification and feature generation models. We expect this model to accelerate the classification of proteins from primary sequence data and increase the accessibility of protein characteristic prediction to a broader range of scientists. m-NGSG is freely available at Bitbucket: https://bitbucket.org/sm_islam/mngsg/src. A web server is available at watson.ecs.baylor.edu/ngsg. erich_baker@baylor.edu. Supplementary data are available at Bioinformatics online.

Agrobacterium-mediated transformation of modified antifreeze protein gene in strawberry

Directory of Open Access Journals (Sweden)

Srisulak Dheeranupattana

2005-07-01

Full Text Available The optimum condition for shoot regeneration from leaf explants of strawberry cultivar Tiogar was investigated. It was found that the best regeneration condition was MS medium containing N6-Benzyladenine (BA and 2,4-Dichlorophenoxy acetic acid (2,4-D at concentrations of 1 mg.l-1 and 0.2 mg.l-1, respectively. Antibiotics sensitivity test found that shoot regeneration from leaf explant was inhibited more than 90% at the concentration of kanamycin (Km as low as 5 mg.l-1. The modified gene encoding antifreeze protein isoform HPLC 6 was successfully constructed using codons which were optimally expressed in the strawberry plant. The antifreeze protein genes, naturally in plasmid pSW1 and modified in plasmid BB, were transformed to strawberry leaf explants by Agrobacterium tumefaciens LBA 4404. The strawberry plants, transformed with both AFP genes, were able to root in MS media containing 50 mg.l-1 Km, while no roots grew from nontransformed plant in this condition. Polymerase chain reaction indicated that the transgenes were integrated in the genome of transformants.

Preparation of Modified Films with Protein from Grouper Fish

Directory of Open Access Journals (Sweden)

M. A. Valdivia-López

2016-01-01

Full Text Available A protein concentrate (PC was obtained from Grouper fish skin and it was used to prepare films with different amounts of sorbitol and glycerol as plasticizers. The best performing films regarding resistance were then modified with various concentrations of CaCl2, CaSO4 (calcium salts, and glucono-δ-lactone (GDL with the purpose of improving their mechanical and barrier properties. These films were characterized by determining their mechanical properties and permeability to water vapor and oxygen. Formulations with 5% (w/v protein and 75% sorbitol and 4% (w/v protein with a mixture of 15% glycerol and 15% sorbitol produced adequate films. Calcium salts and GDL increased the tensile fracture stress but reduced the fracture strain and decreased water vapor permeability compared with control films. The films prepared represent an attractive alternative for being used as food packaging materials.

Charge dynamics in conducting polyaniline–metal oxalate composites

Indian Academy of Sciences (India)

Unknown

Polyaniline; metal oxalate composites; charge transport; mobile and fixed spins; VRH conduc- tion mechanism ... Al, Mn and Co on doping into Pani improve the poly- merization ... dopants on charge dynamics with EPR and other tech- niques.

Effects of insecticidal crystal proteins (Cry proteins) produced by genetically modified maize (Bt maize) on the nematode Caenorhabditis elegans

International Nuclear Information System (INIS)

Höss, Sebastian; Menzel, Ralph; Gessler, Frank; Nguyen, Hang T.; Jehle, Johannes A.; Traunspurger, Walter

2013-01-01

The genetically modified maize MON89034 × MON88017 expresses different crystal (Cry) proteins with pesticidal activity against the European corn borer (Cry1.105; Cry2Ab2) and the Western corn root worm (Cry3Bb1). Non-target organisms, such as soil nematodes, might be exposed to the Cry proteins that enter the soil in course of crop growing. Therefore, the risk of those proteins for nematodes was assessed by testing their toxic effects on Caenorhabditis elegans. All three insecticidal Cry proteins showed dose-dependent inhibitory effects on C. elegans reproduction (EC50: 0.12–0.38 μmol L −1 ), however, at concentrations that were far above the expected soil concentrations. Moreover, a reduced toxicity was observed when Cry proteins were added jointly. A C. elegans mutant strain deficient for receptors for the nematicidal Cry5B was also resistant against Cry1.105 and Cry2Ab2, suggesting that these Cry proteins bound to the same or similar receptors as nematicidal Cry proteins and thereby affect the reproduction of C. elegans. -- Highlights: •Insecticidal Cry proteins dose-dependently inhibited the reproduction of C. elegans. •Mixture toxicity was lower than expected from concentration-additive single effects. •Genes for MAPK-defense-pathway were up-regulated in presence of Cry protein mixture. •Knock-out strains deficient for Cry5B-receptors showed lower susceptibility to insecticidal Cry proteins. •Toxicity of insecticidal Cry-proteins on C. elegans occurred at concentrations far above expected field concentrations. -- Insecticidal Cry proteins expressed by genetically modified maize act on nematodes via a similar mode of action as nematicidal Cry proteins, however, at concentrations far above expected soil levels

Electrostatic contribution of surface charge residues to the stability of a thermophilic protein: benchmarking experimental and predicted pKa values.

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Chi-Ho Chan

Full Text Available Optimization of the surface charges is a promising strategy for increasing thermostability of proteins. Electrostatic contribution of ionizable groups to the protein stability can be estimated from the differences between the pKa values in the folded and unfolded states of a protein. Using this pKa-shift approach, we experimentally measured the electrostatic contribution of all aspartate and glutamate residues to the stability of a thermophilic ribosomal protein L30e from Thermococcus celer. The pKa values in the unfolded state were found to be similar to model compound pKas. The pKa values in both the folded and unfolded states obtained at 298 and 333 K were similar, suggesting that electrostatic contribution of ionizable groups to the protein stability were insensitive to temperature changes. The experimental pKa values for the L30e protein in the folded state were used as a benchmark to test the robustness of pKa prediction by various computational methods such as H++, MCCE, MEAD, pKD, PropKa, and UHBD. Although the predicted pKa values were affected by crystal contacts that may alter the side-chain conformation of surface charged residues, most computational methods performed well, with correlation coefficients between experimental and calculated pKa values ranging from 0.49 to 0.91 (p<0.01. The changes in protein stability derived from the experimental pKa-shift approach correlate well (r = 0.81 with those obtained from stability measurements of charge-to-alanine substituted variants of the L30e protein. Our results demonstrate that the knowledge of the pKa values in the folded state provides sufficient rationale for the redesign of protein surface charges leading to improved protein stability.

Brown pigment formation in heated sugar-protein mixed suspensions containing unmodified and peptically modified whey protein concentrates.

Science.gov (United States)

Rongsirikul, Narumol; Hongsprabhas, Parichat

2016-01-01

Commercial whey protein concentrate (WPC) was modified by heating the acidified protein suspensions (pH 2.0) at 80 °C for 30 min and treating with pepsin at 37 °C for 60 min. Prior to spray-drying, such modification did not change the molecular weights (MWs) of whey proteins determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). After spray-drying the modified whey protein concentrate with trehalose excipient (MWPC-TH), it was found that the α-lactalbumin (α-La) was the major protein that was further hydrolyzed the most. The reconstituted MWPC-TH contained β-lactoglobulin (β-Lg) as the major protein and small molecular weight (MW) peptides of less than 6.5 kDa. The reconstituted MWPC-TH had higher NH2 group, Trolox equivalent antioxidant capacity (TEAC), lower exposed aromatic ring and thiol (SH) contents than did the commercial WPC. Kinetic studies revealed that the addition of MWPC-TH in fructose-glycine solution was able to reduce brown pigment formation in the mixtures heated at 80 to 95 °C by increasing the activation energy (Ea) of brown pigment formation due to the retardation of fluoresced advanced glycation end product (AGEs) formation. The addition of MWPC to reducing sugar-glycine/commercial WPC was also able to lower brown pigment formation in the sterilized (121 °C, 15 min) mixed suspensions containing 0.1 M reducing sugar and 0.5-1.0 % glycine and/or commercial (P < 0.05). It was demonstrated that the modification investigated in this study selectively hydrolyzed α-La and retained β-Lg for the production of antibrowning whey protein concentrate.

Plasma membrane translocation of a protein needle based on a triple-stranded β-helix motif.

Science.gov (United States)

Sanghamitra, Nusrat J M; Inaba, Hiroshi; Arisaka, Fumio; Ohtan Wang, Dan; Kanamaru, Shuji; Kitagawa, Susumu; Ueno, Takafumi

2014-10-01

Plasma membrane translocation is challenging due to the barrier of the cell membrane. Contrary to the synthetic cell-penetrating materials, tailed bacteriophages use cell-puncturing protein needles to puncture the cell membranes as an initial step of the DNA injection process. Cell-puncturing protein needles are thought to remain functional in the native phages. In this paper, we found that a bacteriophage T4 derived protein needle of 16 nm length spontaneously translocates through the living cell membrane. The β-helical protein needle (β-PN) internalizes into human red blood cells that lack endocytic machinery. By comparing the cellular uptake of β-PNs with modified surface charge, it is shown that the uptake efficiency is maximum when it has a negative charge corresponding to a zeta potential value of -16 mV. In HeLa cells, uptake of β-PN incorporates endocytosis independent mechanisms with partial macropinocytosis dependence. The endocytosis dependence of the uptake increases when the surface charges of β-PNs are modified to positive or negative. Thus, these results suggest that natural DNA injecting machinery can serve as an inspiration to design new class of cell-penetrating materials with a tailored mechanism.

Protein structure and ionic selectivity in calcium channels: selectivity filter size, not shape, matters.

Science.gov (United States)

Malasics, Attila; Gillespie, Dirk; Nonner, Wolfgang; Henderson, Douglas; Eisenberg, Bob; Boda, Dezso

2009-12-01

Calcium channels have highly charged selectivity filters (4 COO(-) groups) that attract cations in to balance this charge and minimize free energy, forcing the cations (Na(+) and Ca(2+)) to compete for space in the filter. A reduced model was developed to better understand the mechanism of ion selectivity in calcium channels. The charge/space competition (CSC) mechanism implies that Ca(2+) is more efficient in balancing the charge of the filter because it provides twice the charge as Na(+) while occupying the same space. The CSC mechanism further implies that the main determinant of Ca(2+) versus Na(+) selectivity is the density of charged particles in the selectivity filter, i.e., the volume of the filter (after fixing the number of charged groups in the filter). In this paper we test this hypothesis by changing filter length and/or radius (shape) of the cylindrical selectivity filter of our reduced model. We show that varying volume and shape together has substantially stronger effects than varying shape alone with volume fixed. Our simulations show the importance of depletion zones of ions in determining channel conductance calculated with the integrated Nernst-Planck equation. We show that confining the protein side chains with soft or hard walls does not influence selectivity.

Identification of liver protein targets modified by tienilic acid metabolites using a two-dimensional Western blot-mass spectrometry approach

Science.gov (United States)

Methogo, Ruth Menque; Dansette, Patrick M.; Klarskov, Klaus

2007-12-01

A combined approach based on two-dimensional electrophoresis-immuno-blotting and nanoliquid chromatography coupled on-line with electrospray ionization mass spectrometry (nLC-MS/MS) was used to identify proteins modified by a reactive intermediate of tienilic acid (TA). Liver homogenates from rats exposed to TA were fractionated using ultra centrifugation; four fractions were obtained and subjected to 2D electrophoresis. Following transfer to PVDF membranes, modified proteins were visualized after India ink staining, using an anti-serum raised against TA and ECL detection. Immuno-reactive spots were localized on the PVDF membrane by superposition of the ECL image, protein spots of interest were excised, digested on the membrane with trypsin followed by nLC-MS/MS analysis and protein identification. A total of 15 proteins were identified as likely targets modified by a TA reactive metabolite. These include selenium binding protein 2, senescence marker protein SMP-30, adenosine kinase, Acy1 protein, adenosylhomocysteinase, capping protein (actin filament), protein disulfide isomerase, fumarylacetoacetase, arginase chain A, ketohexokinase, proteasome endopeptidase complex, triosephosphate isomerase, superoxide dismutase, dna-type molecular chaperone hsc73 and malate dehydrogenase.

Protein-coated pH-responsive gold nanoparticles: Microwave-assisted synthesis and surface charge-dependent anticancer activity

Directory of Open Access Journals (Sweden)

Dickson Joseph

2014-09-01

Full Text Available The biocompatibility and ease of functionalization of gold nanoparticles underlie significant potential in biotechnology and biomedicine. Eight different proteins were examined in the preparation of gold nanoparticles (AuNPs in aqueous medium under microwave irradiation. Six of the proteins resulted in the formation of AuNPs. The intrinsic pH of the proteins played an important role in AuNPs with strong surface plasmon bands. The hydrodynamic size of the nanoparticles was larger than the values observed by TEM and ImageJ. The formation of a protein layer on the AuNPs accounts for this difference. The AuNPs exhibited sensitivity towards varying pH conditions, which was confirmed by determining the difference in the isoelectric points studied by using pH-dependent zeta potential titration. Cytotoxicity studies revealed anticancerous effects of the AuNPs at a certain micromolar concentration by constraining the growth of cancer cells with different efficacies due to the use of different proteins as capping agents. The positively charged AuNPs are internalized by the cells to a greater level than the negatively charged AuNPs. These AuNPs synthesized with protein coating holds promise as anticancer agents and would help in providing a new paradigm in area of nanoparticles.

Effect of nonuniform radial density distribution on the space charge dominated beam bunching

International Nuclear Information System (INIS)

Sing Babu, P.; Goswami, A.; Pandit, V. S.

2011-01-01

Beam dynamics of a space charge dominated beam during the bunch compression is studied self consistently for the case of fixed shape non-uniform bell shape and hollow shape density distributions in the transverse direction. We have used thick slices at different parts of the beam to account for variation in the beam radius in the study of the transverse dynamics. The longitudinal dynamics has been studied using the disc model. The axial variation of the radius of the slices and emittance growth arising from the phase dependence of the transverse rf forces are also included in the simulation. We have modified the beam envelope equation to take into account the longitudinal space charge effect on the transverse motion which arises due to the finite bunch size. To demonstrate the application of the theoretical formulations developed, we have studied a sinusoidal beam bunching system and presented detailed numerical results.

Role of uniform pore structure and high positive charges in the arsenate adsorption performance of Al13-modified montmorillonite

International Nuclear Information System (INIS)

Zhao, Shou; Feng, Chenghong; Huang, Xiangning; Li, Baohua; Niu, Junfeng; Shen, Zhenyao

2012-01-01

Highlights: ► Al 13 modification changes As(V) sorption mechanism of montmorillonites. ► Intercalated ion charges mainly affects As(V) adsorption kinetics. ► Uniform pore structure exhibit more excellent As(V) adsorption performance. - Abstract: Four modified montmorillonite adsorbents with varied Al 13 contents (i.e., Na-Mont, AC-Mont, PAC 20 -Mont, and Al 13 -Mont) were synthesized and characterized by N 2 adsorption/desorption, X-ray diffraction, and Fourier-transform infrared analyses. The arsenate adsorption performance of the four adsorbents were also investigated to determine the role of intercalated Al 13 , especially its high purity, high positive charge (+7), and special Keggin structure. With increased Al 13 content, the physicochemical properties (e.g., surface area, structural uniformity, basal spacing, and pore volume) and adsorption performance of the modified montmorillonites were significantly but disproportionately improved. The adsorption data well fitted the Freundlich and Redlich–Peterson isotherm model, whereas the kinetic data better correlated with the pseudo-second-order kinetic model. The arsenate sorption mechanism of the montmorillonites changed from physical to chemisorption after intercalation with Al 13 . Increasing charges of the intercalated ions enhanced the arsenate adsorption kinetics, but had minimal effect on the structural changes of the montmorillonites. The uniform pore structure formed by intercalation with high-purity Al 13 greatly enhanced the pore diffusion and adsorption rate of arsenate, resulting in the high adsorption performance of Al 13 -Mont.

Non-isochronous spiral orbit particle accelerator and fixed frequency closed orbit particle accelerator

International Nuclear Information System (INIS)

Fujisawa, Takashi; Hattori, Toshiyuki

2006-01-01

One of the present inventions provides a spiral orbit charged particle accelerator in which the magnetic field increases as the radius increases more rapidly than an isochronous magnetic field distribution, and the distribution of fixed-frequency accelerating RF voltage is formed so that a harmonic number changes in integer for every particle revolution. The other invention realizes to make the closed orbit charged particle accelerator having a fixed frequency amplitude modulator that is able to modulate amplitude of the RF voltage so that a harmonic number decreases in integer in an every particle revolution. (author)

Charge-Discharge Properties of the Surface-Modified ZrNi Alloy Electrode with Different Degrees of Boiling Alkaline Treatment

Directory of Open Access Journals (Sweden)

Akihiro Matsuyama

2016-09-01

Full Text Available Charge-discharge properties of the surface-modified ZrNi negative electrodes with different degrees of boiling alkaline treatment were investigated. The boiling alkaline treatment was performed by immersing the ZrNi electrode in a boiling 6 M KOH aqueous solution for 2 h or 4 h. The initial discharge capacity for the untreated ZrNi negative electrode was 21 mAh·g−1, but it was increased to 114 mAh·g−1 and 308 mAh·g−1 after the boiling alkaline treatments for 2 h and 4 h, respectively. The discharge capacity for the ZrNi negative electrode after the treatment for 2 h steadily increased with repeating charge-discharge cycles as well as that of the untreated electrode, whereas that for the ZrNi negative electrode after the 4 h treatment greatly decreased. The high rate of dischargeability was improved with an increase in the treatment period of time, and the charge-transfer resistance was drastically decreased. Scanning electron microscopy (SEM and electron dispersive X-ray spectroscopy demonstrated the ZrO2 passive layer on the ZrNi alloy surface was removed by the boiling alkaline treatment to form a porous morphology containing Ni(OH2, which can be reduced to Ni during charging, leading to the reduction of a barrier for the charge-discharge reactions.

Spatiotemporal kinetics of γ-H2AX protein on charged particles induced DNA damage

Energy Technology Data Exchange (ETDEWEB)

Niu, H., E-mail: hniu@mx.nthu.edu.tw [Nuclear Science and Technology Development Center, National Tsing Hua University, Hsinchu, Taiwan (China); Chang, H.C. [Department of Biomedical Engineering and Environmental Sciences, National Tsing Hua University, Hsinchu, Taiwan (China); Cho, I.C. [Institute for Radiological Research, Chang Gung University and Chang Gung Memorial Hospital, Taoyuan, Taiwan (China); Chen, C.H. [Nuclear Science and Technology Development Center, National Tsing Hua University, Hsinchu, Taiwan (China); Liu, C.S. [Cancer Center of Taipei Veterans General Hospital, Taipei, Taiwan (China); Chou, W.T. [Department of Biomedical Engineering and Environmental Sciences, National Tsing Hua University, Hsinchu, Taiwan (China)

2014-08-15

Highlights: • Charged particles can induce more complex DNA damages, and these complex damages have higher ability to cause the cell death or cell carcinogenesis. • In this study, we used γ-H2AX protein to investigate the spatiotemporal kinetics of DNA double strand breaks in particle irradiated HeLa cells. • The HeLa cells were irradiated by 400 keV alpha-particles in four different dosages. • The result shows that a good linear relationship can be observed between foci number and radiation dose. • The data shows that the dissolution rate of γ-H2AX foci agree with the two components DNA repairing model, and it was decreasing as the radiation dose increased. • These results suggest that charged particles can induce more complex DNA damages and causing the retardation of DNA repair. - Abstract: In several researches, it has been demonstrated that charged particles can induce more complex DNA damages. These complex damages have higher ability to cause the cell death or cell carcinogenesis. For this reason, clarifying the DNA repair mechanism after charged particle irradiation plays an important role in the development of charged particle therapy and space exploration. Unfortunately, the detail spatiotemporal kinetic of DNA damage repair is still unclear. In this study, we used γ-H2AX protein to investigate the spatiotemporal kinetics of DNA double strand breaks in alpha-particle irradiated HeLa cells. The result shows that the intensity of γ-H2AX foci increased gradually, and reached to its maximum at 30 min after irradiation. A good linear relationship can be observed between foci intensity and radiation dose. After 30 min, the γ-H2AX foci intensity was decreased with time passed, but remained a large portion (∼50%) at 48 h passed. The data show that the dissolution rate of γ-H2AX foci agreed with two components DNA repairing model. These results suggest that charged particles can induce more complex DNA damages and causing the retardation of DNA

Collisional charging of individual submillimeter particles: Using ultrasonic levitation to initiate and track charge transfer

Science.gov (United States)

Lee, Victor; James, Nicole M.; Waitukaitis, Scott R.; Jaeger, Heinrich M.

2018-03-01

Electrostatic charging of insulating fine particles can be responsible for numerous phenomena ranging from lightning in volcanic plumes to dust explosions. However, even basic aspects of how fine particles become charged are still unclear. Studying particle charging is challenging because it usually involves the complexities associated with many-particle collisions. To address these issues, we introduce a method based on acoustic levitation, which makes it possible to initiate sequences of repeated collisions of a single submillimeter particle with a flat plate, and to precisely measure the particle charge in situ after each collision. We show that collisional charge transfer between insulators is dependent on the hydrophobicity of the contacting surfaces. We use glass, which we modify by attaching nonpolar molecules to the particle, the plate, or both. We find that hydrophilic surfaces develop significant positive charges after contacting hydrophobic surfaces. Moreover, we demonstrate that charging between a hydrophilic and a hydrophobic surface is suppressed in an acidic environment and enhanced in a basic one. Application of an electric field during each collision is found to modify the charge transfer, again depending on surface hydrophobicity. We discuss these results within the context of contact charging due to ion transfer, and we show that they lend strong support to O H- ions as the charge carriers.

Modified Mason number for charged paramagnetic colloidal suspensions

Science.gov (United States)

Du, Di; Hilou, Elaa; Biswal, Sibani Lisa

2016-06-01

The dynamics of magnetorheological fluids have typically been described by the Mason number, a governing parameter defined as the ratio between viscous and magnetic forces in the fluid. For most experimental suspensions of magnetic particles, surface forces, such as steric and electrostatic interactions, can significantly influence the dynamics. Here we propose a theory of a modified Mason number that accounts for surface forces and show that this modified Mason number is a function of interparticle distance. We demonstrate that this modified Mason number is accurate in describing the dynamics of a rotating pair of paramagnetic colloids of identical or mismatched sizes in either high or low salt solutions. The modified Mason number is confirmed to be pseudoconstant for particle pairs and particle chains undergoing a stable-metastable transition during rotation. The interparticle distance term can be calculated using theory or can be measured experimentally. This modified Mason number is more applicable to magnetorheological systems where surface forces are not negligible.

Determination of soluble protein contents from RVNRL

International Nuclear Information System (INIS)

Wan Manshol Wan Zin; Nurulhuda Othman

1996-01-01

This project was carried out to determine the soluble protein contents on RVNRL film vulcanisates, with respect to the RVNRL storage time, gamma irradiation dose absorbed by the latex and the effect of different leaching time and leaching conditions. These three factors are important in the hope to determine the best possible mean of minimizing the soluble protein contents in products made from RVNRL. Within the nine months storage period employed in the study, the results show that, the longer the storage period the less the soluble protein extracted from the film samples. Gamma irradiation dose absorbed by the samples, between 5.3 kGy to 25.2 kGy seems to influence the soluble protein contents of the RVNRL films vulcanisates. The higher the dose the more was the soluble protein extracted from the film samples. At an absorbed dose of 5.3 kGy and 25.2 kGy, the soluble contents were 0. 198 mg/ml and 0.247 mg/ml respectively. At a fixed leaching temperature, the soluble proteins increases with leaching time and at a fixed leaching time, the soluble proteins increases with leaching temperature. ne highest extractable protein contents was determined at a leaching time of 10 minutes and leaching temperature of 90'C The protein analysis were done by using Modified Lowry Method

Fixed interface charges between AlGaN barrier and gate stack composed of in situ grown SiN and Al2O3 in AlGaN/GaN high electron mobility transistors with normally off capability

International Nuclear Information System (INIS)

Capriotti, M.; Alexewicz, A.; Fleury, C.; Gavagnin, M.; Bethge, O.; Wanzenböck, H. D.; Bertagnolli, E.; Pogany, D.; Strasser, G.; Visalli, D.; Derluyn, J.

2014-01-01

Using a generalized extraction method, the fixed charge density N int at the interface between in situ deposited SiN and 5 nm thick AlGaN barrier is evaluated by measurements of threshold voltage V th of an AlGaN/GaN metal insulator semiconductor high electron mobility transistor as a function of SiN thickness. The thickness of the originally deposited 50 nm thick SiN layer is reduced by dry etching. The extracted N int is in the order of the AlGaN polarization charge density. The total removal of the in situ SiN cap leads to a complete depletion of the channel region resulting in V th  = +1 V. Fabrication of a gate stack with Al 2 O 3 as a second cap layer, deposited on top of the in situ SiN, is not introducing additional fixed charges at the SiN/Al 2 O 3 interface

Nondissipative optimum charge regulator

Science.gov (United States)

Rosen, R.; Vitebsky, J. N.

1970-01-01

Optimum charge regulator provides constant level charge/discharge control of storage batteries. Basic power transfer and control is performed by solar panel coupled to battery through power switching circuit. Optimum controller senses battery current and modifies duty cycle of switching circuit to maximize current available to battery.

Edible foam based on Pickering effect of probiotic bacteria and milk proteins

DEFF Research Database (Denmark)

Yücel, Cigdem; Geng, Xiaolu; Cárdenas, Marité

2017-01-01

We report the preparation and characterization of aqueous Pickering foams using bio-particles constituted by lactic acid bacteria surface modified by oppositely charged milk proteins. Cell surface modification was shown by zeta potential measurements. Foams stabilized by bacterial Pickering bio-p...

Efficiency of Database Search for Identification of Mutated and Modified Proteins via Mass Spectrometry

OpenAIRE

Pevzner, Pavel A.; Mulyukov, Zufar; Dancik, Vlado; Tang, Chris L

2001-01-01

Although protein identification by matching tandem mass spectra (MS/MS) against protein databases is a widespread tool in mass spectrometry, the question about reliability of such searches remains open. Absence of rigorous significance scores in MS/MS database search makes it difficult to discard random database hits and may lead to erroneous protein identification, particularly in the case of mutated or post-translationally modified peptides. This problem is especially important for high-thr...

Nanoscale charge transfer in redox proteins and DNA: Towards biomolecular electronics

International Nuclear Information System (INIS)

Artés, Juan Manuel; López-Martínez, Montserrat; Díez-Pérez, Ismael; Sanz, Fausto; Gorostiza, Pau

2014-01-01

Understanding how charges move through and between biomolecules is a fundamental question that constitutes the basis for many biological processes. On the other hand, it has potential applications in the design of sensors based on biomolecules and single molecule devices. In this review we introduce the study of the electron transfer (ET) process in biomolecules, providing an overview of the fundamental theory behind it and the different experimental approaches. The ET in proteins is introduced by reviewing a complete electronic characterization of a redox protein (azurin) using electrochemical scanning tunnelling microscopy (ECSTM). The ET process in DNA is overviewed and results from different experimental approaches are discussed. Finally, future directions in the study of the ET process in biomolecules are introduced as well as examples of possible technological applications

Post-duplication charge evolution of phosphoglucose isomerases in teleost fishes through weak selection on many amino acid sites

Directory of Open Access Journals (Sweden)

Sato Yukuto

2007-10-01

Full Text Available Abstract Background The partitioning of ancestral functions among duplicated genes by neutral evolution, or subfunctionalization, has been considered the primary process for the evolution of novel proteins (neofunctionalization. Nonetheless, how a subfunctionalized protein can evolve into a more adaptive protein is poorly understood, mainly due to the limitations of current analytical methods, which can detect only strong selection for amino acid substitutions involved in adaptive molecular evolution. In this study, we employed a comparative evolutionary approach to this question, focusing on differences in the structural properties of a protein, specifically the electric charge, encoded by fish-specific duplicated phosphoglucose isomerase (Pgi genes. Results Full-length cDNA cloning, RT-PCR based gene expression analyses, and comparative sequence analyses showed that after subfunctionalization with respect to the expression organ of duplicate Pgi genes, the net electric charge of the PGI-1 protein expressed mainly in internal tissues became more negative, and that of PGI-2 expressed mainly in muscular tissues became more positive. The difference in net protein charge was attributable not to specific amino acid sites but to the sum of various amino acid sites located on the surface of the PGI molecule. Conclusion This finding suggests that the surface charge evolution of PGI proteins was not driven by strong selection on individual amino acid sites leading to permanent fixation of a particular residue, but rather was driven by weak selection on a large number of amino acid sites and consequently by steady directional and/or purifying selection on the overall structural properties of the protein, which is derived from many modifiable sites. The mode of molecular evolution presented here may be relevant to various cases of adaptive modification in proteins, such as hydrophobic properties, molecular size, and electric charge.

Fixed-Target Electron Accelerators

International Nuclear Information System (INIS)

Brooks, William K.

2001-01-01

A tremendous amount of scientific insight has been garnered over the past half-century by using particle accelerators to study physical systems of sub-atomic dimensions. These giant instruments begin with particles at rest, then greatly increase their energy of motion, forming a narrow trajectory or beam of particles. In fixed-target accelerators, the particle beam impacts upon a stationary sample or target which contains or produces the sub-atomic system being studied. This is in distinction to colliders, where two beams are produced and are steered into each other so that their constituent particles can collide. The acceleration process always relies on the particle being accelerated having an electric charge; however, both the details of producing the beam and the classes of scientific investigations possible vary widely with the specific type of particle being accelerated. This article discusses fixed-target accelerators which produce beams of electrons, the lightest charged particle. As detailed in the report, the beam energy has a close connection with the size of the physical system studied. Here a useful unit of energy is a GeV, i.e., a giga electron-volt. (ne GeV, the energy an electron would have if accelerated through a billion volts, is equal to 1.6 x 10 -10 joules.) To study systems on a distance scale much smaller than an atomic nucleus requires beam energies ranging from a few GeV up to hundreds of GeV and more

STM and transport measurements of highly charged ion modified materials

International Nuclear Information System (INIS)

Pomeroy, J.M.; Grube, H.; Perrella, A.C.; Gillaspy, J.D.

2007-01-01

Careful measurements of highly charged ions (HCIs) colliding with gases and surfaces have provided glimpses of intense electronic interactions, but a comprehensive model for the interaction mechanisms, time scales, and resultant nano-features that bridges materials systems is yet to be realized. At the National Institute of Standards and Technology (NIST) electron beam ion trap (EBIT) facility, new apparatus is now connected to the HCI beamline to allow preparation of clean, atomically flat surfaces of single crystals, e.g. gold, tungsten and silicon, and deposition and patterning of thin films, e.g. high resistivity oxides, ferromagnetic metals, normal metals and superconductors. Experiments reported here focus on the electronic and morphological structure of HCI induced nano-features. Current activities are focused on using in situ scanning tunneling microscope (STM) on Au(1 1 1) and (separately) ex situ transport measurements to study electronic properties within HCI modified magnetic multilayer systems. Specifically, we are fabricating magnetic multilayers similar to magnetic tunnel junctions (MTJs) (important in advanced magnetic field sensors and superconducting Josephson junction devices) and using HCIs to adjust critical electronic properties. The electrical response of the tunnel junction to HCIs provides a novel approach to performing HCI-induced nanostructure ensemble measurements

Measurement of SEU thresholds and cross sections at fixed incidence angles

International Nuclear Information System (INIS)

Criswell, T.L.; Oberg, D.L.; Wert, J.L.; Measel, P.R.; Wilson, W.E.

1987-01-01

Current SEU testing and analysis techniques have as basic assumptions that the charge deposited at a junction depends linearly on the linear energy transfer (LET) of the ion and the pathlength of the ion through an imagined parallelepiped that represents the depletion region. This study tests these assumptions for two bipolar parts, AMD 27LS00 and Fairchild 93L422, by irradiating at fixed angles while varying the LET of two ion species. It was found that the 27LS00 shows a pronounced ion species dependence, and may show a deviation of deposited charge from the usual inverse-cosine times a fixed depletion depth, while the 93L422 exhibited the expected inverse-cosine dependence and no ion species dependence

Size-matched alkyne-conjugated cyanine fluorophores to identify differences in protein glycosylation.

Science.gov (United States)

Burnham-Marusich, Amanda R; Plechaty, Anna M; Berninsone, Patricia M

2014-09-01

Currently, there are few methods to detect differences in posttranslational modifications (PTMs) in a specific manner from complex mixtures. Thus, we developed an approach that combines the sensitivity and specificity of click chemistry with the resolution capabilities of 2D-DIGE. In "Click-DIGE", posttranslationally modified proteins are metabolically labeled with azido-substrate analogs, then size- and charge-matched alkyne-Cy3 or alkyne-Cy5 dyes are covalently attached to the azide of the PTM by click chemistry. The fluorescently-tagged protein samples are then multiplexed for 2DE analysis. Whereas standard DIGE labels all proteins, Click-DIGE focuses the analysis of protein differences to a targeted subset of posttranslationally modified proteins within a complex sample (i.e. specific labeling and analysis of azido glycoproteins within a cell lysate). Our data indicate that (i) Click-DIGE specifically labels azido proteins, (ii) the resulting Cy-protein conjugates are spectrally distinct, and (iii) the conjugates are size- and charge-matched at the level of 2DE. We demonstrate the utility of this approach by detecting multiple differentially expressed glycoproteins between a mutant cell line defective in UDP-galactose transport and the parental cell line. We anticipate that the diversity of azido substrates already available will enable Click-DIGE to be compatible with analysis of a wide range of PTMs. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Use of carbamylated charge standards for testing batches of ampholytes used in two-dimensional elecrophoresis

Energy Technology Data Exchange (ETDEWEB)

Tollaksen, S L; Edwards, J J; Anderson, N G

1981-01-01

A method of testing batches of ampholytes is presented. By using carbamylated charge standards to co-electrophorese with the protein sample in the first-dimension isoelectric focusing gel, one can monitor, after running and staining the second-dimension sodium dodecyl sulfate (SDS) slab gel, the continuity of the pH gradient. Charge standards can also be used to check the reproducibility of the pH gradient among batches of ampholytes and to modify the new batch with a small amount of a narrow range ampholyte to assure reproducibility of experiments. Ampholytes for comparison were obtained from three major manufacturers. 5 figures.

Electronic transport in single-helical protein molecules: Effects of multiple charge conduction pathways and helical symmetry

Energy Technology Data Exchange (ETDEWEB)

Kundu, Sourav, E-mail: sourav.kunduphy@gmail.com; Karmakar, S.N.

2016-07-15

We propose a tight-binding model to investigate electronic transport properties of single helical protein molecules incorporating both the helical symmetry and the possibility of multiple charge transfer pathways. Our study reveals that due to existence of both the multiple charge transfer pathways and helical symmetry, the transport properties are quite rigid under influence of environmental fluctuations which indicates that these biomolecules can serve as better alternatives in nanoelectronic devices than its other biological counterparts e.g., single-stranded DNA.

Charge-exchange collisions of multiply charged ions with atoms

International Nuclear Information System (INIS)

Grozdanov, T.P.; Janev, R.K.

1978-01-01

The problem of electron transfer between neutral atoms and multiply charged ions is considered at low and medium energies. It is assumed that a large number of final states are available for the electron transition so that the electron-capture process is treated as a tunnel effect caused by the strong attractive Coulomb field of the multicharged ions. The electron transition probability is obtained in a closed form using the modified-comparison-equation method to solve the Schroedinger equation. An approximately linear dependence of the one-electron transfer cross section on the charge of multicharged ion is found. Cross-section calculations of a number of charge-exchange reactions are performed

Extension of the self-consistent-charge density-functional tight-binding method: third-order expansion of the density functional theory total energy and introduction of a modified effective coulomb interaction.

Science.gov (United States)

Yang, Yang; Yu, Haibo; York, Darrin; Cui, Qiang; Elstner, Marcus

2007-10-25

The standard self-consistent-charge density-functional-tight-binding (SCC-DFTB) method (Phys. Rev. B 1998, 58, 7260) is derived by a second-order expansion of the density functional theory total energy expression, followed by an approximation of the charge density fluctuations by charge monopoles and an effective damped Coulomb interaction between the atomic net charges. The central assumptions behind this effective charge-charge interaction are the inverse relation of atomic size and chemical hardness and the use of a fixed chemical hardness parameter independent of the atomic charge state. While these approximations seem to be unproblematic for many covalently bound systems, they are quantitatively insufficient for hydrogen-bonding interactions and (anionic) molecules with localized net charges. Here, we present an extension of the SCC-DFTB method to incorporate third-order terms in the charge density fluctuations, leading to chemical hardness parameters that are dependent on the atomic charge state and a modification of the Coulomb scaling to improve the electrostatic treatment within the second-order terms. These modifications lead to a significant improvement in the description of hydrogen-bonding interactions and proton affinities of biologically relevant molecules.

Novel Prostate Specific Antigen plastic antibody designed with charged binding sites for an improved protein binding and its application in a biosensor of potentiometric transduction

International Nuclear Information System (INIS)

Rebelo, Tânia S.C.R.; Santos, C.; Costa-Rodrigues, J.; Fernandes, M.H.; Noronha, João P.; Sales, M. Goreti F.

2014-01-01

Graphical abstract: EF13-201, Novel Prostate Specific Antigen plastic antibody designed with charged binding sites for an improved protein binding and its application in a biosensor of potentiometric transduction. - Abstract: This work shows that the synthesis of protein plastic antibodies tailored with selected charged monomers around the binding site enhances protein binding. These charged receptor sites are placed over a neutral polymeric matrix, thus inducing a suitable orientation the protein reception to its site. This is confirmed by preparing control materials with neutral monomers and also with non-imprinted template. This concept has been applied here to Prostate Specific Antigen (PSA), the protein of choice for screening prostate cancer throughout the population, with serum levels >10 ng/mL pointing out a high probability of associated cancer. Protein Imprinted Materials with charged binding sites (C/PIM) have been produced by surface imprinting over graphene layers to which the protein was first covalently attached. Vinylbenzyl(trimethylammonium chloride) and vinyl benzoate were introduced as charged monomers labelling the binding site and were allowed to self-organize around the protein. The subsequent polymerization was made by radical polymerization of vinylbenzene. Neutral PIM (N/PIM) prepared without oriented charges and non imprinted materials (NIM) obtained without template were used as controls. These materials were used to develop simple and inexpensive potentiometric sensor for PSA. They were included as ionophores in plasticized PVC membranes, and tested over electrodes of solid or liquid conductive contacts, made of conductive carbon over a syringe or of inner reference solution over micropipette tips. The electrodes with charged monomers showed a more stable and sensitive response, with an average slope of -44.2 mV/decade and a detection limit of 5.8 × 10 −11 mol/L (2 ng/mL). The corresponding non-imprinted sensors showed lower

Oxidatively Modified Proteins in the Serous Subtype of Ovarian Carcinoma

Directory of Open Access Journals (Sweden)

Sharifeh Mehrabi

2014-01-01

Full Text Available Serous subtype of ovarian cancer is considered to originate from fallopian epithelium mucosa that has been exposed to physiological changes resulting from ovulation. Ovulation influences an increased in inflammation of epithelial ovarian cells as results of constant exposure of cells to ROS. The imbalance between ROS and antioxidant capacities, as well as a disruption of redox signaling, causes a wide range of damage to DNA, proteins, and lipids. This study applied spectrophotometric, dinitrophenylhydrazone (DNPH assay, two-dimensional gel electrophoresis, and Western blot analyses to assess the levels of oxidatively modified proteins in 100 primary serous epithelial ovarian carcinoma and normal/surrounding tissues. These samples were obtained from 56 Caucasian and 44 African-American patients within the age range of 61±10 years. Analyses showed that the levels of reactive protein carbonyl groups increased as stages progressed to malignancy. Additionally, the levels of protein carbonyls in serous ovarian carcinoma among African Americans are 40% (P<0.05 higher relative to Caucasian at similar advanced stages. Results suggest that oxidative stress is involved in the modification of carbonyl protein groups, leading to increased aggressiveness of epithelial ovarian tumors and may contribute to the disease's invasiveness among African Americans.

Dielectric spectra of proteins in conducting media

International Nuclear Information System (INIS)

Ruderman, G.; Xammar Oro, J.R. de

1990-10-01

Dielectric measurements of serum albumin and myoglobin in solutions of varying conductivities were performed. The results presented confirm that also for protein solutions, the Maxwell predictions of a threshold frequency in conducting materials holds. The threshold frequency of a serum albumin solution was experimentally determined. Attention should be recalled that, if the dielectric spectra of proteins solutions want to be measured, three distinct frequency regions are to be observed: a low frequency region, where the sample behaves like a conductor; an intermediate region centered around the threshold frequency, where the free charges partially screen the fixed ones; and a high frequency region where the sample behaves like a good dielectric. (author). 8 refs, 5 figs

Chromatographic and traditional albumin isotherms on cellulose: a model for wound protein adsorption on modified cotton

Science.gov (United States)

Albumin is the most abundant protein found in healing wounds. Traditional and chromatogrpahic protein isotherms of albumin binding on modified cotton fibers are useful in understanding albumin binding to cellulose wound dressings. An important consideration in the design of cellulosic wound dressin...

Studies of the aggregation of mutant proteins in vitro provide insights into the genetics of amyloid diseases.

Science.gov (United States)

Chiti, Fabrizio; Calamai, Martino; Taddei, Niccolo; Stefani, Massimo; Ramponi, Giampietro; Dobson, Christopher M

2002-12-10

Protein aggregation and the formation of highly insoluble amyloid structures is associated with a range of debilitating human conditions, which include Alzheimer's disease, Parkinson's disease, and the Creutzfeldt-Jakob disease. Muscle acylphosphatase (AcP) has already provided significant insights into mutational changes that modulate amyloid formation. In the present paper, we have used this system to investigate the effects of mutations that modify the charge state of a protein without affecting significantly the hydrophobicity or secondary structural propensities of the polypeptide chain. A highly significant inverse correlation was found to exist between the rates of aggregation of the protein variants under denaturing conditions and their overall net charge. This result indicates that aggregation is generally favored by mutations that bring the net charge of the protein closer to neutrality. In light of this finding, we have analyzed natural mutations associated with familial forms of amyloid diseases that involve alteration of the net charge of the proteins or protein fragments associated with the diseases. Sixteen mutations have been identified for which the mechanism of action that causes the pathological condition is not yet known or fully understood. Remarkably, 14 of these 16 mutations cause the net charge of the corresponding peptide or protein that converts into amyloid deposits to be reduced. This result suggests that charge has been a key parameter in molecular evolution to ensure the avoidance of protein aggregation and identifies reduction of the net charge as an important determinant in at least some forms of protein deposition diseases.

Quantitative analysis of modified proteins and their positional isomers by tandem mass spectrometry: human histone H4.

Science.gov (United States)

Pesavento, James J; Mizzen, Craig A; Kelleher, Neil L

2006-07-01

Here we show that fragment ion abundances from dissociation of ions created from mixtures of multiply modified histone H4 (11 kDa) or of N-terminal synthetic peptides (2 kDa) correspond to their respective intact ion abundances measured by Fourier transform mass spectrometry. Isomeric mixtures of modified forms of the same protein are resolved and quantitated with a precision of protein ions created by electrospray greatly easing many of the systematic biases that more strongly affect small peptides (e.g., differences in ionization efficiency and ion m/z values). The ion fragmentation methods validated here are directly extensible to intact human proteins to derive quantitative information on the highly related and often isomeric protein forms created by combinatorial arrays of posttranslational modifications.

On the configuration of an active target for a fixed-target B experiment at SSC energies

International Nuclear Information System (INIS)

Dukes, E.C.

1993-01-01

The optimal configuration of target and silicon microvertex detector for fixed-target B experiments has yet to be determined. For fixed-target charm experiments the usual setup consists of a series of inert target foils - typically a few millimeters thick and separated by a few centimeters - immediately followed by a silicon microvertex detector. Because of the larger boost at the SSC, the efficacy of using active target foils - tightly packed silicon microstrip detectors - has been considered by at least one group: the SFT collaboration. It is hoped that with an active target the tracks of charged B's themselves can be measured, improving charged B reconstruction efficiencies. The author examines two issues concerning silicon active targets for fixed-target experiments at the SSC: (1) the effect on the acceptance of the requirement that the B decay vertices occur outside of the target foils, and (2) the ability of an active target to directly track charged B's

Simple protein structure-sensitive chronopotentiometric analysis with dithiothreitol-modified Hg electrodes

Czech Academy of Sciences Publication Activity Database

Ostatná, Veronika; Černocká, Hana; Paleček, Emil

2012-01-01

Roč. 87, SI (2012), s. 84-88 ISSN 1567-5394 R&D Projects: GA AV ČR(CZ) KJB100040901; GA ČR(CZ) GAP301/11/2055; GA MŠk(CZ) LC06035 Institutional research plan: CEZ:AV0Z50040507; CEZ:AV0Z50040702 Keywords : protein electroanalysis * DTT-modified electrodes * electrocatalysis Subject RIV: BO - Biophysics Impact factor: 3.947, year: 2012

NTERACTION BETWEEN SURFACE CHARGE PHENOMENA AND MULTI-SPECIES DIFFUSION IN CEMENT BASED MATERIALS

DEFF Research Database (Denmark)

Johannesson, Björn

2008-01-01

Measurements strongly indicate that the ‘inner’ surface of the microscopic structure of cement based materials has a fixed negative charge. This charge contributes to the formation of so-called electrical double layers. In the case of cement based materials the ionic species located in such layers...... are typically potassium -, sodium - and calcium ions. Due to the high specific surface area of hydrated cement, a large amount of ions can be located in theses double layers even if the surface charge is relatively low. The attraction force, caused by the fixed surface charge on ions located close to surfaces......, is one possible explanation for the observed low global diffusion rates in the pore system of positively charged ions compared to the negatively charged ones. Here it is of interest to simulate the multi ionic diffusion behavior when assigning positively charged ions a comparably lower diffusion constant...

Hydrogen-peroxide-modified egg albumen for transparent and flexible resistive switching memory

Science.gov (United States)

Zhou, Guangdong; Yao, Yanqing; Lu, Zhisong; Yang, Xiude; Han, Juanjuan; Wang, Gang; Rao, Xi; Li, Ping; Liu, Qian; Song, Qunliang

2017-10-01

Egg albumen is modified by hydrogen peroxide with concentrations of 5%, 10%, 15% and 30% at room temperature. Compared with devices without modification, a memory cell of Ag/10% H2O2-egg albumen/indium tin oxide exhibits obviously enhanced resistive switching memory behavior with a resistance ratio of 104, self-healing switching endurance for 900 cycles and a prolonged retention time for a 104 s @ 200 mV reading voltage after being bent 103 times. The breakage of massive protein chains occurs followed by the recombination of new protein chain networks due to the oxidation of amidogen and the synthesis of disulfide during the hydrogen peroxide modifying egg albumen. Ions such as Fe3+, Na+, K+, which are surrounded by protein chains, are exposed to the outside of protein chains to generate a series of traps during the egg albumen degeneration process. According to the fitting results of the double logarithm I-V curves and the current-sensing atomic force microscopy (CS-AFM) images of the ON and OFF states, the charge transfer from one trap center to its neighboring trap center is responsible for the resistive switching memory phenomena. The results of our work indicate that hydrogen- peroxide-modified egg albumen could open up a new avenue of biomaterial application in nanoelectronic systems.

ESIprot: a universal tool for charge state determination and molecular weight calculation of proteins from electrospray ionization mass spectrometry data.

Science.gov (United States)

Winkler, Robert

2010-02-01

Electrospray ionization (ESI) ion trap mass spectrometers with relatively low resolution are frequently used for the analysis of natural products and peptides. Although ESI spectra of multiply charged protein molecules also can be measured on this type of devices, only average spectra are produced for the majority of naturally occurring proteins. Evaluating such ESI protein spectra would provide valuable information about the native state of investigated proteins. However, no suitable and freely available software could be found which allows the charge state determination and molecular weight calculation of single proteins from average ESI-MS data. Therefore, an algorithm based on standard deviation optimization (scatter minimization) was implemented for the analysis of protein ESI-MS data. The resulting software ESIprot was tested with ESI-MS data of six intact reference proteins between 12.4 and 66.7 kDa. In all cases, the correct charge states could be determined. The obtained absolute mass errors were in a range between -0.2 and 1.2 Da, the relative errors below 30 ppm. The possible mass accuracy allows for valid conclusions about the actual condition of proteins. Moreover, the ESIprot algorithm demonstrates an extraordinary robustness and allows spectral interpretation from as little as two peaks, given sufficient quality of the provided m/z data, without the necessity for peak intensity data. ESIprot is independent from the raw data format and the computer platform, making it a versatile tool for mass spectrometrists. The program code was released under the open-source GPLv3 license to support future developments of mass spectrometry software. Copyright 2010 John Wiley & Sons, Ltd.

Multiplicity distributions of charged hadrons in vp and charged current interactions

Science.gov (United States)

Jones, G. T.; Jones, R. W. L.; Kennedy, B. W.; Morrison, D. R. O.; Mobayyen, M. M.; Wainstein, S.; Aderholz, M.; Hantke, D.; Katz, U. F.; Kern, J.; Schmitz, N.; Wittek, W.; Borner, H. P.; Myatt, G.; Radojicic, D.; Burke, S.

1992-03-01

Using data on vp andbar vp charged current interactions from a bubble chamber experiment with BEBC at CERN, the multiplicity distributions of charged hadrons are investigated. The analysis is based on ˜20000 events with incident v and ˜10000 events with incidentbar v. The invariant mass W of the total hadronic system ranges from 3 GeV to ˜14 GeV. The experimental multiplicity distributions are fitted by the binomial function (for different intervals of W and in different intervals of the rapidity y), by the Levy function and the lognormal function. All three parametrizations give acceptable values for X 2. For fixed W, forward and backward multiplicities are found to be uncorrelated. The normalized moments of the charged multiplicity distributions are measured as a function of W. They show a violation of KNO scaling.

Theory and measurements of electrophoretic effects in monolith, fixed-bed, and fluidized-bed plasma reactors

International Nuclear Information System (INIS)

Morin, T.J.

1989-01-01

Pressure gradients and secondary flow fields generated by the passage of electrical current in a d.c. gas discharge or gas laser are topics of longstanding interest in the gaseous electronics literature. These hydrodynamic effects of space charge fields and charged particle density gradients have been principally exploited in the development of gas separation and purification processes. In recent characterization studies of fixed-bed and fluidized-bed plasma reactors several anomalous flow features have been observed. These reactors involve the contacting of a high-frequency, resonantly-sustained, disperse gas discharge with granular solids in a fixed or fluidized bed. Anomalies in the measured pressure drops and fluidization velocities have motivated the development of an appropriate theoretical approach to, and some additional experimental investigations of electrophoretic effects in disperse gas discharges. In this paper, a theory which includes the effects of space charge and diffusion is used to estimate the electric field and charged particle density profiles. These profiles are then used to calculate velocity fields and gas flow rates for monolith, fixed-bed, and fluidized-bed reactors. These results are used to rationalize measurements of gas flow rates and axial pressure gradients in high-frequency disperse gas discharges with and without an additional d.c. axial electric field

Vertical orbit excursion fixed field alternating gradient accelerators

Directory of Open Access Journals (Sweden)

Stephen Brooks

2013-08-01

Full Text Available Fixed field alternating gradient (FFAG accelerators with vertical orbit excursion (VFFAGs provide a promising alternative design for rings with fixed-field superconducting magnets. They have a vertical magnetic field component that increases with height in the vertical aperture, yielding a skew quadrupole focusing structure. Scaling-type VFFAGs are found with fixed tunes and no intrinsic limitation on momentum range. This paper presents the first multiparticle tracking of such machines. Proton driver rings to accelerate the 800 MeV beam from the ISIS synchrotron are presented, in terms of both magnet field geometry and longitudinal behavior during acceleration with space charge. The 12 GeV ring produces an output power of at least 2.18 MW. Possible applications of VFFAGs to waste transmutation, hadron therapy, and energy-recovery electron accelerators are also discussed.

Protein arrangement on modified diamond-like carbon surfaces – An ARXPS study

Energy Technology Data Exchange (ETDEWEB)

Oosterbeek, Reece N., E-mail: reece.oosterbeek@auckland.ac.nz [Department of Chemical and Materials Engineering, The University of Auckland, Private Bag 92019 (New Zealand); Seal, Christopher K. [Light Metals Research Centre, The University of Auckland, Private Bag 92019 (New Zealand); Hyland, Margaret M. [Department of Chemical and Materials Engineering, The University of Auckland, Private Bag 92019 (New Zealand)

2014-12-01

Highlights: • DLC coatings were modified by Ar{sup +} ion sputtering and laser graphitisation. • The surface properties of the coatings were measured, and it was found that the above methods increased sp{sup 2} content and altered surface energy. • ARXPS was used to observe protein arrangement on the surface. • Polar CO/CN groups were seen to be segregated towards the interface, indicating they play an important role in bonding. • This segregation increased with increasing polar surface energy, indicating an increased net attraction between polar groups. - Abstract: Understanding the nature of the interface between a biomaterial implant and the biological fluid is an essential step towards creating improved implant materials. This study examined a diamond-like carbon coating biomaterial, the surface energy of which was modified by Ar{sup +} ion sputtering and laser graphitisation. The arrangement of proteins was analysed by angle resolved X-ray photoelectron spectroscopy, and the effects of the polar component of surface energy on this arrangement were observed. It was seen that polar groups (such as CN, CO) are more attracted to the coating surface due to the stronger polar interactions. This results in a segregation of these groups to the DLC–protein interface; at increasing takeoff angle (further from to DLC–protein interface) fewer of these polar groups are seen. Correspondingly, groups that interact mainly by dispersive forces (CC, CH) were found to increase in intensity as takeoff angle increased, indicating they are segregated away from the DLC–protein interface. The magnitude of the segregation was seen to increase with increasing polar surface energy, this was attributed to an increased net attraction between the solid surface and polar groups at higher polar surface energy (γ{sub S}{sup p})

Protein arrangement on modified diamond-like carbon surfaces – An ARXPS study

International Nuclear Information System (INIS)

Oosterbeek, Reece N.; Seal, Christopher K.; Hyland, Margaret M.

2014-01-01

Highlights: • DLC coatings were modified by Ar + ion sputtering and laser graphitisation. • The surface properties of the coatings were measured, and it was found that the above methods increased sp 2 content and altered surface energy. • ARXPS was used to observe protein arrangement on the surface. • Polar CO/CN groups were seen to be segregated towards the interface, indicating they play an important role in bonding. • This segregation increased with increasing polar surface energy, indicating an increased net attraction between polar groups. - Abstract: Understanding the nature of the interface between a biomaterial implant and the biological fluid is an essential step towards creating improved implant materials. This study examined a diamond-like carbon coating biomaterial, the surface energy of which was modified by Ar + ion sputtering and laser graphitisation. The arrangement of proteins was analysed by angle resolved X-ray photoelectron spectroscopy, and the effects of the polar component of surface energy on this arrangement were observed. It was seen that polar groups (such as CN, CO) are more attracted to the coating surface due to the stronger polar interactions. This results in a segregation of these groups to the DLC–protein interface; at increasing takeoff angle (further from to DLC–protein interface) fewer of these polar groups are seen. Correspondingly, groups that interact mainly by dispersive forces (CC, CH) were found to increase in intensity as takeoff angle increased, indicating they are segregated away from the DLC–protein interface. The magnitude of the segregation was seen to increase with increasing polar surface energy, this was attributed to an increased net attraction between the solid surface and polar groups at higher polar surface energy (γ S p )

Precise Point Positioning with Partial Ambiguity Fixing.

Science.gov (United States)

Li, Pan; Zhang, Xiaohong

2015-06-10

Reliable and rapid ambiguity resolution (AR) is the key to fast precise point positioning (PPP). We propose a modified partial ambiguity resolution (PAR) method, in which an elevation and standard deviation criterion are first used to remove the low-precision ambiguity estimates for AR. Subsequently the success rate and ratio-test are simultaneously used in an iterative process to increase the possibility of finding a subset of decorrelated ambiguities which can be fixed with high confidence. One can apply the proposed PAR method to try to achieve an ambiguity-fixed solution when full ambiguity resolution (FAR) fails. We validate this method using data from 450 stations during DOY 021 to 027, 2012. Results demonstrate the proposed PAR method can significantly shorten the time to first fix (TTFF) and increase the fixing rate. Compared with FAR, the average TTFF for PAR is reduced by 14.9% for static PPP and 15.1% for kinematic PPP. Besides, using the PAR method, the average fixing rate can be increased from 83.5% to 98.2% for static PPP, from 80.1% to 95.2% for kinematic PPP respectively. Kinematic PPP accuracy with PAR can also be significantly improved, compared to that with FAR, due to a higher fixing rate.

Distribution network connection pricing framework and methodology: identification of areas of improvement for Sarawak Energy Berhad Connection Charges Guidelines through modified delphi method

Science.gov (United States)

Tan, J. K.; Abas, N.

2017-07-01

Complaints on issues and matters related to connection charges have been very common for electricity supply utility companies around the world including Sarawak Energy Berhad. In order to identify the areas that can be improved, a mixed method of exploratory research involving qualitative and quantitative methods have been designed and undertaken rather than a single method of survey. This will ensure a more comprehensive and detailed understanding of the issues from various target groups. The method is designed under three phases, employing Modified Delphi Technique for phase 1 through a series of stake holder engagements, online and offline survey questionnaires to be filled by internal wiring contractors for phase 2 whilst under phase 3, case studies shall be carried out on the issues identified from phase 1 and phase 2 of the study. This paper presented the findings from the Modified Delphi Technique. The findings revealed that there are areas of improvement for Sarawak Energy Berhad connection guidelines in term of differentiation of dedicated and shared assets which leads to unfairness to the connecting customers, inconsistency and non-transparent in charging. The findings of Modified Delphi Technique shall be used for implementation of phase 2 and phase 3 of the study.

Importance of Accurate Charges in Binding Affinity Calculations: A Case of Neuraminidase Series

Energy Technology Data Exchange (ETDEWEB)

Park, Kichul; Kyun, Nack Sung; Cho, Art E. [Korea Univ., Sejong (Korea, Republic of)

2013-02-15

It has been shown that calculating atomic charges using quantum mechanical level theory greatly improves the accuracy of docking. A protocol was developed and shown to be effective. That this protocol works is just a manifestation of the fact that electrostatic interactions are important in protein-ligand binding. In order to investigate how the same protocol helps in prediction of binding affinities, we took a series of known cocrystal structures of influenza neuraminidase inhibitors with the receptor and performed docking with Glide SP, Glide XP, and QPLD, the last being a workflow that incorporates QM/MM calculations to replace the fixed atomic charges of force fields with quantum mechanically recalculated ones at a given docking pose, and predicted the binding affinities of each cocrystal. The correlation with experimental binding affinities considerably improved with QPLD compared to Glide SP/XP yielding r{sup 2} = 0.83. The results suggest that for binding sites, such as that of neuraminidase, which are laden with hydrophilic residues, protocols such as QPLD which utilizes QM-based atomic charges can better predict the binding affinities.

Pyrenebutanoate as a dynamic protein modifier for fluorometric detection in capillary zone electrophoresis

Czech Academy of Sciences Publication Activity Database

Horká, Marie; Šlais, Karel

2002-01-01

Roč. 23, 7-8 (2002), s. 1090-1095 ISSN 0173-0835 R&D Projects: GA AV ČR IAA4031901 Institutional research plan: CEZ:AV0Z4031919 Keywords : pyrenebutanoate * dynamic protein modifier * CZE Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 4.325, year: 2002

Serum lipids modify periodontal infection - C-reactive protein association.

Science.gov (United States)

Haro, Anniina; Saxlin, Tuomas; Suominen, Anna-Liisa; Ylöstalo, Pekka; Leiviskä, Jaana; Tervonen, Tellervo; Knuuttila, Matti

2012-09-01

To investigate whether low-grade inflammation-related factors such as serum low-density (LDL-C) and high-density lipoprotein cholesterol (HDL-C) modify the association between periodontal infection and C-reactive protein. This study was based on a subpopulation of the Health 2000 Survey, which consisted of dentate, non-diabetic, non-rheumatic subjects who were 30-49 years old (n = 2710). The extent of periodontal infection was measured by means of the number of teeth with periodontal pocket ≥4 mm and teeth with periodontal pocket ≥6 mm and systemic inflammation using high sensitive C-reactive protein. The extent of periodontal infection was associated with elevated levels of C-reactive protein among those subjects whose HDL-C value was below the median value of 1.3 mmol/l or LDL-C above the median value of 3.4 mmol/l. Among those with HDL-C ≥ 1.3 mmol/l or LDL-C ≤ 3.4 mmol/l, the association between periodontal infection and serum concentrations of C-reactive protein was practically non-existent. This study suggests that the relation of periodontal infection to the systemic inflammatory condition is more complicated than previously presumed. The findings of this study suggest that the possible systemic effect of periodontal infection is dependent on serum lipid composition. © 2012 John Wiley & Sons A/S.

STUDY OF HYDRODYNAMICS IN FIXED BED OF COMPOSITE GRANULAR MATERIALS

Directory of Open Access Journals (Sweden)

Stelian Petrescu

2010-12-01

Full Text Available This study aims at the experimental determination of pressure drop and friction factor at gas flow through fixed beds of granular silica gel, alumina and activated carbon, and establishment of an equation containing a modified friction factor Fm to calculate pressure drop. In order to calculate the modified friction factor, an equation was suggested.The experimental values for pressure drop and friction factor were determined using spherical grains of silica gel, cylindrical grains of alumina and silica gel, alumina and activated carbon impregnated with calcium chloride. By means of the suggested equation, the values of pressure drop in fixed bed were calculated and compared with the experimental values. A good agreement between the predicted and experimental data is noticed.

O-GlcNAc-specific antibody CTD110.6 cross-reacts with N-GlcNAc2-modified proteins induced under glucose deprivation.

Directory of Open Access Journals (Sweden)

Takahiro Isono

Full Text Available Modification of serine and threonine residues in proteins by O-linked β-N-acetylglucosamine (O-GlcNAc glycosylation is a feature of many cellular responses to the nutritional state and to stress. O-GlcNAc modification is reversibly regulated by O-linked β-N-acetylglucosamine transferase (OGT and β-D-N-acetylglucosaminase (O-GlcNAcase. O-GlcNAc modification of proteins is dependent on the concentration of uridine 5'-diphospho-N-acetylglucosamine (UDP-GlcNAc, which is a substrate of OGT and is synthesized via the hexosamine biosynthetic pathway. Immunoblot analysis using the O-GlcNAc-specific antibody CTD110.6 has indicated that glucose deprivation increases protein O-GlcNAcylation in some cancer cells. The mechanism of this paradoxical phenomenon has remained unclear. Here we show that the increased glycosylation induced by glucose deprivation and detected by CTD110.6 antibodies is actually modification by N-GlcNAc(2, rather than by O-GlcNAc. We found that this induced glycosylation was not regulated by OGT and O-GlcNAcase, unlike typical O-GlcNAcylation, and it was inhibited by treatment with tunicamycin, an N-glycosylation inhibitor. Proteomics analysis showed that proteins modified by this induced glycosylation were N-GlcNAc(2-modified glycoproteins. Furthermore, CTD110.6 antibodies reacted with N-GlcNAc(2-modified glycoproteins produced by a yeast strain with a ts-mutant of ALG1 that could not add a mannose residue to dolichol-PP-GlcNAc(2. Our results demonstrated that N-GlcNAc(2-modified glycoproteins were induced under glucose deprivation and that they cross-reacted with the O-GlcNAc-specific antibody CTD110.6. We therefore propose that the glycosylation status of proteins previously classified as O-GlcNAc-modified proteins according to their reactivity with CTD110.6 antibodies must be re-examined. We also suggest that the repression of mature N-linked glycoproteins due to increased levels of N-GlcNAc(2-modified proteins is a newly

[Attaching single- and multi-unit fixed dental prostheses

NARCIS (Netherlands)

Kreulen, C.M.; Wolke, J.G.C.; Baat, C. de; Creugers, N.H.J.

2013-01-01

A single- or multi-unit fixed dental prosthesis can be attached to the abutment teeth through mechanical retention and gap sealing or by adhesion. For sealing the gap, water-soluble cements are appropriate, such as zinc phosphate, polycarboxylate, and (resin-modified) glasionomer cement. Attachment

Integrated and convenient procedure for protein extraction from formalin-fixed, paraffin-embedded tissues for LC-MS/MS analysis.

Science.gov (United States)

Lai, Xianyin; Schneider, Bryan P

2014-11-01

Because fresh-frozen tissue samples associated with long-term clinical data and of rare diseases are often unobtainable at the present time, formalin-fixed paraffin-embedded (FFPE) tissue samples are considered a highly valuable resource for researchers. However, protein extraction from FFPE tissues faces challenges of deparaffinization and cross-link reversion. Current procedures for protein extraction from FFPE tissue require separate steps and toxic solvents, resulting in inconvenience in protein extraction. To overcome these limitations, an integrated method was developed using nontoxic solvents in four types of FFPE tissues. The average amount of proteins from three replicates of bladder, kidney, liver, and lung FFPE tissues were 442.6, 728.9, 736.4, and 694.7 μg with CVs of 7.5, 5.8, 2.4, and 4.5%, respectively. Proteomic analysis showed that 348, 417, 607, and 304 unique proteins were identified and quantified without specification of isoform by a least two peptides from bladder, kidney, liver, and lung FFPE tissue samples, respectively. The analysis of individual protein CV demonstrated that 97-99% of the proteins were quantified with a CV ≤ 30%, verifying the reproducibility of the integrated protein extraction method. In summary, the developed method is high-yield, reproducible, convenient, simple, low cost, nonvolatile, nonflammable, and nontoxic. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Proteomic workflow for analysis of archival formalin-fixed and paraffin-embedded clinical samples to a depth of 10 000 proteins.

Science.gov (United States)

Wiśniewski, Jacek R; Duś, Kamila; Mann, Matthias

2013-04-01

Archival formalin-fixed and paraffin-embedded clinical samples represent a very diverse source of material for proteomic investigation of diseases, often with follow-up patient information. Here, we describe an analytical workflow for analysis of laser-capture microdissected formalin-fixed and paraffin-embedded samples that allows studying proteomes to a depth of 10 000 proteins per sample. The workflow involves lysis of tissue in SDS-containing buffer, detergent removal, and consecutive digestion of the proteins with two enzymes by the multienzyme digestion filter-aided sample preparation method. Resulting peptides are fractionated by pipette-tip based strong anion exchange into six fractions and analyzed by LC-MS/MS on a bench top quadrupole Orbitrap mass spectrometer. Analysis of the data using the MaxQuant software resulted in the identification of 9502 ± 28 protein groups per a 110 nL sample of microdissected cells from human colonic adenoma. This depth of proteome analysis enables systemic insights into the organization of the adenoma cells and an estimation of the abundances of known biomarkers. It also allows the identification of proteins expressed from tumor suppressors, oncogenes, and other key players in the development and progression of the colorectal cancer. Our proteomic platform can be used for quantitative comparisons between samples representing different stages of diseases and thus can be applied to the discovery of biomarkers or drug targets. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Interactions between Therapeutic Proteins and Acrylic Acid Leachable.

Science.gov (United States)

Liu, Dengfeng; Nashed-Samuel, Yasser; Bondarenko, Pavel V; Brems, David N; Ren, Da

2012-01-01

Leachables are chemical compounds that migrate from manufacturing equipment, primary containers and closure systems, and packaging components into biopharmaceutical and pharmaceutical products. Acrylic acid (at concentration around 5 μg/mL) was detected as leachable in syringes from one of the potential vendors (X syringes). In order to evaluate the potential impact of acrylic acid on therapeutic proteins, an IgG 2 molecule was filled into a sterilized X syringe and then incubated at 45 °C for 45 days in a pH 5 acetate buffer. We discovered that acrylic acid can interact with proteins at three different sites: (1) the lysine side chain, (2) the N-terminus, and (3) the histidine side chain, by the Michael reaction. In this report, the direct interactions between acrylic acid leachable and a biopharmaceutical product were demonstrated and the reaction mechanism was proposed. Even thought a small amount (from 0.02% to 0.3%) of protein was found to be modified by acrylic acid, the modified protein can potentially be harmful due to the toxicity of acrylic acid. After being modified by acrylic acid, the properties of the therapeutic protein may change due to charge and hydrophobicity variations. Acrylic acid was detected to migrate from syringes (Vendor X) into a therapeutic protein solution (at a concentration around 5 μg/mL). In this study, we discovered that acrylic acid can modify proteins at three different sites: (1) the lysine side chain, 2) the N-terminus, and 3) the histidine side chain, by the Michael reaction. In this report, the direct interactions between acrylic acid leachable and a biopharmaceutical product were demonstrated and the reaction mechanism was proposed.

Receptor Activity-modifying Protein-directed G Protein Signaling Specificity for the Calcitonin Gene-related Peptide Family of Receptors.

Science.gov (United States)

Weston, Cathryn; Winfield, Ian; Harris, Matthew; Hodgson, Rose; Shah, Archna; Dowell, Simon J; Mobarec, Juan Carlos; Woodlock, David A; Reynolds, Christopher A; Poyner, David R; Watkins, Harriet A; Ladds, Graham

2016-10-14

The calcitonin gene-related peptide (CGRP) family of G protein-coupled receptors (GPCRs) is formed through the association of the calcitonin receptor-like receptor (CLR) and one of three receptor activity-modifying proteins (RAMPs). Binding of one of the three peptide ligands, CGRP, adrenomedullin (AM), and intermedin/adrenomedullin 2 (AM2), is well known to result in a Gα s -mediated increase in cAMP. Here we used modified yeast strains that couple receptor activation to cell growth, via chimeric yeast/Gα subunits, and HEK-293 cells to characterize the effect of different RAMP and ligand combinations on this pathway. We not only demonstrate functional couplings to both Gα s and Gα q but also identify a Gα i component to CLR signaling in both yeast and HEK-293 cells, which is absent in HEK-293S cells. We show that the CGRP family of receptors displays both ligand- and RAMP-dependent signaling bias among the Gα s , Gα i , and Gα q/11 pathways. The results are discussed in the context of RAMP interactions probed through molecular modeling and molecular dynamics simulations of the RAMP-GPCR-G protein complexes. This study further highlights the importance of RAMPs to CLR pharmacology and to bias in general, as well as identifying the importance of choosing an appropriate model system for the study of GPCR pharmacology. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

SU(3) lattice gauge fixing with overrelaxation and Gribov copies

Energy Technology Data Exchange (ETDEWEB)

Paciello, M.L.; Taglienti, B. (INFN La Sapienza, Rome (Italy)); Parrinello, C. (Physics Dept., New York Univ., NY (United States)); Petrarca, S. (Theory Div., CERN, Geneva (Switzerland)); Vladikas, A. (Dipt. di Fisica, Univ. Tor Vergata, Rome (Italy) INFN Tor Vergata, Rome (Italy))

1992-02-06

We report on the phenomenology of SU(3) lattice Landau gauge fixing as obtained by using an overrelaxation algorithm. An interesting result obtained using this very efficient algorithm is that distinct Gribov copies are generated by simply modifying the value {omega} of the overrelaxation parameter for a fixed starting configuration. By generating random gauge equivalent configurations, we study the variation of the number of copies with the lattice volume and gauge coupling. (orig.).

Modifying Poisson equation for near-solute dielectric polarization and solvation free energy

Energy Technology Data Exchange (ETDEWEB)

Yang, Pei-Kun, E-mail: peikun@isu.edu.tw

2016-06-15

Highlights: • We modify the Poisson equation. • The dielectric polarization was calculated from the modified Poisson equation. • The solvation free energies of the solutes were calculated from the dielectric polarization. • The calculated solvation free energies were similar to those obtained from MD simulations. - Abstract: The dielectric polarization P is important for calculating the stability of protein conformation and the binding affinity of protein–protein/ligand interactions and for exploring the nonthermal effect of an external electric field on biomolecules. P was decomposed into the product of the electric dipole moment per molecule p; bulk solvent density N{sub bulk}; and relative solvent molecular density g. For a molecular solute, 4πr{sup 2}p(r) oscillates with the distance r to the solute, and g(r) has a large peak in the near-solute region, as observed in molecular dynamics (MD) simulations. Herein, the Poisson equation was modified for computing p based on the modified Gauss’s law of Maxwell’s equations, and the potential of the mean force was used for computing g. For one or two charged atoms in a water cluster, the solvation free energies of the solutes obtained by these equations were similar to those obtained from MD simulations.

The Role of Conformational Flexibility on Protein Supercharging in Native Electrospray Ionization

Science.gov (United States)

Sterling, Harry J.; Cassou, Catherine A.; Trnka, Michael J.; Burlingame, A. L.; Krantz, Bryan A.; Williams, Evan R.

2012-01-01

Effects of covalent intramolecular bonds, either native disulfide bridges or chemical crosslinks, on ESI supercharging of proteins from aqueous solutions were investigated. Chemically modifying cytochrome c with up to seven crosslinks or ubiquitin with up to two crosslinks did not affect the average or maximum charge states of these proteins in the absence of m-nitrobenzyl alcohol (m-NBA), but the extent of supercharging induced by m-NBA increased with decreasing numbers of crosslinks. For the model random coil polypeptide reduced/alkylated RNase A, a decrease in charging with increasing m-NBA concentration attributable to reduced surface tension of the ESI droplet was observed, whereas native RNase A electrosprayed from these same solutions exhibited enhanced charging. The inverse relationship between the extent of supercharging and the number of intramolecular crosslinks for folded proteins, as well as the absence of supercharging for proteins that are random coils in aqueous solution, indicate that conformational restrictions induced by the crosslinks reduce the extent of supercharging. These results provide additional evidence that protein and protein complex supercharging from aqueous solution is primarily due to partial or significant unfolding that occurs as a result of chemical and/or thermal denaturation induced by the supercharging reagent late in the ESI droplet lifetime. PMID:21399817

Elevated pressure improves the extraction and identification of proteins recovered from formalin-fixed, paraffin-embedded tissue surrogates.

Directory of Open Access Journals (Sweden)

Carol B Fowler

2010-12-01

Full Text Available Proteomic studies of formalin-fixed paraffin-embedded (FFPE tissues are frustrated by the inability to extract proteins from archival tissue in a form suitable for analysis by 2-D gel electrophoresis or mass spectrometry. This inability arises from the difficulty of reversing formaldehyde-induced protein adducts and cross-links within FFPE tissues. We previously reported the use of elevated hydrostatic pressure as a method for efficient protein recovery from a hen egg-white lysozyme tissue surrogate, a model system developed to study formalin fixation and histochemical processing.In this study, we demonstrate the utility of elevated hydrostatic pressure as a method for efficient protein recovery from FFPE mouse liver tissue and a complex multi-protein FFPE tissue surrogate comprised of hen egg-white lysozyme, bovine carbonic anhydrase, bovine ribonuclease A, bovine serum albumin, and equine myoglobin (55∶15∶15∶10∶5 wt%. Mass spectrometry of the FFPE tissue surrogates retrieved under elevated pressure showed that both the low and high-abundance proteins were identified with sequence coverage comparable to that of the surrogate mixture prior to formaldehyde treatment. In contrast, non-pressure-extracted tissue surrogate samples yielded few positive and many false peptide identifications. Studies with soluble formalin-treated bovine ribonuclease A demonstrated that pressure modestly inhibited the rate of reversal (hydrolysis of formaldehyde-induced protein cross-links. Dynamic light scattering studies suggest that elevated hydrostatic pressure and heat facilitate the recovery of proteins free of formaldehyde adducts and cross-links by promoting protein unfolding and hydration with a concomitant reduction in the average size of the protein aggregates.These studies demonstrate that elevated hydrostatic pressure treatment is a promising approach for improving the recovery of proteins from FFPE tissues in a form suitable for proteomic analysis.

Elevated Pressure Improves the Extraction and Identification of Proteins Recovered from Formalin-Fixed, Paraffin-Embedded Tissue Surrogates

Science.gov (United States)

Fowler, Carol B.; Chesnick, Ingrid E.; Moore, Cedric D.; O'Leary, Timothy J.; Mason, Jeffrey T.

2010-01-01

Background Proteomic studies of formalin-fixed paraffin-embedded (FFPE) tissues are frustrated by the inability to extract proteins from archival tissue in a form suitable for analysis by 2-D gel electrophoresis or mass spectrometry. This inability arises from the difficulty of reversing formaldehyde-induced protein adducts and cross-links within FFPE tissues. We previously reported the use of elevated hydrostatic pressure as a method for efficient protein recovery from a hen egg-white lysozyme tissue surrogate, a model system developed to study formalin fixation and histochemical processing. Principal Findings In this study, we demonstrate the utility of elevated hydrostatic pressure as a method for efficient protein recovery from FFPE mouse liver tissue and a complex multi-protein FFPE tissue surrogate comprised of hen egg-white lysozyme, bovine carbonic anhydrase, bovine ribonuclease A, bovine serum albumin, and equine myoglobin (55∶15∶15∶10∶5 wt%). Mass spectrometry of the FFPE tissue surrogates retrieved under elevated pressure showed that both the low and high-abundance proteins were identified with sequence coverage comparable to that of the surrogate mixture prior to formaldehyde treatment. In contrast, non-pressure-extracted tissue surrogate samples yielded few positive and many false peptide identifications. Studies with soluble formalin-treated bovine ribonuclease A demonstrated that pressure modestly inhibited the rate of reversal (hydrolysis) of formaldehyde-induced protein cross-links. Dynamic light scattering studies suggest that elevated hydrostatic pressure and heat facilitate the recovery of proteins free of formaldehyde adducts and cross-links by promoting protein unfolding and hydration with a concomitant reduction in the average size of the protein aggregates. Conclusions These studies demonstrate that elevated hydrostatic pressure treatment is a promising approach for improving the recovery of proteins from FFPE tissues in a form

Protein arrangement on modified diamond-like carbon surfaces - An ARXPS study

Science.gov (United States)

Oosterbeek, Reece N.; Seal, Christopher K.; Hyland, Margaret M.

2014-12-01

Understanding the nature of the interface between a biomaterial implant and the biological fluid is an essential step towards creating improved implant materials. This study examined a diamond-like carbon coating biomaterial, the surface energy of which was modified by Ar+ ion sputtering and laser graphitisation. The arrangement of proteins was analysed by angle resolved X-ray photoelectron spectroscopy, and the effects of the polar component of surface energy on this arrangement were observed. It was seen that polar groups (such as CN, CO) are more attracted to the coating surface due to the stronger polar interactions. This results in a segregation of these groups to the DLC-protein interface; at increasing takeoff angle (further from to DLC-protein interface) fewer of these polar groups are seen. Correspondingly, groups that interact mainly by dispersive forces (CC, CH) were found to increase in intensity as takeoff angle increased, indicating they are segregated away from the DLC-protein interface. The magnitude of the segregation was seen to increase with increasing polar surface energy, this was attributed to an increased net attraction between the solid surface and polar groups at higher polar surface energy (γSp).

Shotgun proteomic analytical approach for studying proteins adsorbed onto liposome surface

KAUST Repository

Capriotti, Anna Laura

2011-07-02

The knowledge about the interaction between plasma proteins and nanocarriers employed for in vivo delivery is fundamental to understand their biodistribution. Protein adsorption onto nanoparticle surface (protein corona) is strongly affected by vector surface characteristics. In general, the primary interaction is thought to be electrostatic, thus surface charge of carrier is supposed to play a central role in protein adsorption. Because protein corona composition can be critical in modifying the interactive surface that is recognized by cells, characterizing its formation onto lipid particles may serve as a fundamental predictive model for the in vivo efficiency of a lipidic vector. In the present work, protein coronas adsorbed onto three differently charged cationic liposome formulations were compared by a shotgun proteomic approach based on nano-liquid chromatography-high-resolution mass spectrometry. About 130 proteins were identified in each corona, with only small differences between the different cationic liposome formulations. However, this study could be useful for the future controlled design of colloidal drug carriers and possibly in the controlled creation of biocompatible surfaces of other devices that come into contact with proteins into body fluids. © 2011 Springer-Verlag.

Screening for Glycosylphosphatidylinositol-Modified Cell Wall Proteins in Pichia pastoris and Their Recombinant Expression on the Cell Surface

Science.gov (United States)

Zhang, Li; Liang, Shuli; Zhou, Xinying; Jin, Zi; Jiang, Fengchun; Han, Shuangyan; Zheng, Suiping

2013-01-01

Glycosylphosphatidylinositol (GPI)-anchored glycoproteins have various intrinsic functions in yeasts and different uses in vitro. In the present study, the genome of Pichia pastoris GS115 was screened for potential GPI-modified cell wall proteins. Fifty putative GPI-anchored proteins were selected on the basis of (i) the presence of a C-terminal GPI attachment signal sequence, (ii) the presence of an N-terminal signal sequence for secretion, and (iii) the absence of transmembrane domains in mature protein. The predicted GPI-anchored proteins were fused to an alpha-factor secretion signal as a substitute for their own N-terminal signal peptides and tagged with the chimeric reporters FLAG tag and mature Candida antarctica lipase B (CALB). The expression of fusion proteins on the cell surface of P. pastoris GS115 was determined by whole-cell flow cytometry and immunoblotting analysis of the cell wall extracts obtained by β-1,3-glucanase digestion. CALB displayed on the cell surface of P. pastoris GS115 with the predicted GPI-anchored proteins was examined on the basis of potential hydrolysis of p-nitrophenyl butyrate. Finally, 13 proteins were confirmed to be GPI-modified cell wall proteins in P. pastoris GS115, which can be used to display heterologous proteins on the yeast cell surface. PMID:23835174

Experimental determination of net protein charge, [A]tot, and Ka of nonvolatile buffers in bird plasma.

Science.gov (United States)

Stämpfli, Henry; Taylor, Michael; McNicoll, Carl; Gancz, Ady Y; Constable, Peter D

2006-06-01

The quantitative mechanistic acid-base approach to clinical assessment of acid-base status requires species-specific values for [A]tot (the total concentration of nonvolatile buffers in plasma) and Ka (the effective dissociation constant for weak acids in plasma). The aim of this study was to determine [A]tot and Ka values for plasma in domestic pigeons. Plasma from 12 healthy commercial domestic pigeons was tonometered with 20% CO2 at 37 degrees C. Plasma pH, Pco2, and plasma concentrations of strong cations (Na, K, Ca), strong anions (Cl, L-lactate), and nonvolatile buffer ions (total protein, albumin, phosphate) were measured over a pH range of 6.8-7.7. Strong ion difference (SID) (SID5=Na+K+Ca-Cl-lactate) was used to calculate [A]tot and Ka from the measured pH and Pco2 and SID5. Mean (+/-SD) values for bird plasma were as follows: [A]tot=7.76+/-2.15 mmol/l (equivalent to 0.32 mmol/g of total protein, 0.51 mmol/g of albumin, 0.23 mmol/g of total solids); Ka=2.15+/-1.15x10(-7); and pKa=6.67. The net protein charge at normal pH (7.43) was estimated to be 6 meq/l; this value indicates that pigeon plasma has a much lower anion gap value than mammals after adjusting for high mean L-lactate concentrations induced by restraint during blood sampling. This finding indicates that plasma proteins in pigeons have a much lower net anion charge than mammalian plasma protein. An incidental finding was that total protein concentration measured by a multianalyzer system was consistently lower than the value for total solids measured by refractometer.

THE VALENCE OF CORPUSCULAR PROTEINS.

Science.gov (United States)

Gorin, M H; Mover, L S

1942-07-20

BY THE USE OF TWO EXTREME MODELS: a hydrated sphere and an unhydrated rod the valence (net charge) of corpuscular proteins can be successfully calculated from electric mobility data by the Debye-Hückel theory (modified to include the effect of the ions in the ion atmosphere) in conjunction with the electrophoretic theory of Henry. As pointed out by Abramson, this permits a comparison with values for the valence from titration data. Electrometric titration measurements of serum albumin B (Kekwick) have been determined at several ionic strengths. These results, together with the available data in the literature for serum albumin B, egg albumin, and beta-lactoglobulin have been used to compare values for the valence calculated from measurements of titration, electrophoresis, and membrane potentials. The results indicate that the usual interpretation of titration curves is open to serious question. By extrapolation of the titration data to zero ionic strength and protein concentration, there results an "intrinsic" net charge curve describing the binding of H(+) (OH(-)) ion alone. This curve agrees closely, in each case, with values of the valence calculated from mobility data (which in turn are in close accord with those estimated from membrane potential measurements). The experimental titration curves in the presence of appreciable quantities of ions and protein deviate widely from the ideal curve. It is suggested that, under these conditions, binding of undissociated acid (base) leads to erroneous values for the net charge. This binding would not affect the electrophoretic mobility. Values of the net charge obtained by the two extreme models from electrophoretic data are in agreement within 15 to 20 per cent. The agreement between the cylindrical model and the titration data is somewhat better in each case than with the sphere; i.e., this comparison enables a choice to be made between asymmetry and hydration in the interpretation of results from sedimentation and

Model for screened, charge-regulated electrostatics of an eye lens protein: Bovine gammaB-crystallin

Science.gov (United States)

Wahle, Christopher W.; Martini, K. Michael; Hollenbeck, Dawn M.; Langner, Andreas; Ross, David S.; Hamilton, John F.; Thurston, George M.

2017-09-01

We model screened, site-specific charge regulation of the eye lens protein bovine gammaB-crystallin (γ B ) and study the probability distributions of its proton occupancy patterns. Using a simplified dielectric model, we solve the linearized Poisson-Boltzmann equation to calculate a 54 ×54 work-of-charging matrix, each entry being the modeled voltage at a given titratable site, due to an elementary charge at another site. The matrix quantifies interactions within patches of sites, including γ B charge pairs. We model intrinsic p K values that would occur hypothetically in the absence of other charges, with use of experimental data on the dependence of p K values on aqueous solution conditions, the dielectric model, and literature values. We use Monte Carlo simulations to calculate a model grand-canonical partition function that incorporates both the work-of-charging and the intrinsic p K values for isolated γ B molecules and we calculate the probabilities of leading proton occupancy configurations, for 4 Debye screening lengths from 6 to 20 Å. We select the interior dielectric value to model γ B titration data. At p H 7.1 and Debye length 6.0 Å, on a given γ B molecule the predicted top occupancy pattern is present nearly 20% of the time, and 90% of the time one or another of the first 100 patterns will be present. Many of these occupancy patterns differ in net charge sign as well as in surface voltage profile. We illustrate how charge pattern probabilities deviate from the multinomial distribution that would result from use of effective p K values alone and estimate the extents to which γ B charge pattern distributions broaden at lower p H and narrow as ionic strength is lowered. These results suggest that for accurate modeling of orientation-dependent γ B -γ B interactions, consideration of numerous pairs of proton occupancy patterns will be needed.

Improved Interaction Potentials for Charged Residues in Proteins

DEFF Research Database (Denmark)

Kepp, Kasper Planeta

2008-01-01

Electrostatic interactions dominate the structure and free energy of biomolecules. To obtain accurate free energies involving charged groups from molecular simulations, OPLS-AA parameters have been reoptimized using Monte Carlo free energy perturbation. New parameters fit a self-consistent, exper......Electrostatic interactions dominate the structure and free energy of biomolecules. To obtain accurate free energies involving charged groups from molecular simulations, OPLS-AA parameters have been reoptimized using Monte Carlo free energy perturbation. New parameters fit a self......, TIP4P or TIP3P; i.e., each water model requires specific water-charged molecule interaction potentials. New models (models 1 and 3) are thus described for both water models. Uncertainties in relative free energies of charged residues are ~2 kcal/mol with the new parameters, due to variations in system...

Vertical detachment energy of hydrated electron based on a modified form of solvent reorganization energy.

Science.gov (United States)

Wang, Xing-Jian; Zhu, Quan; Li, Yun-Kui; Cheng, Xue-Min; Li, Xiang-Yuan; Fu, Ke-Xiang; He, Fu-Cheng

2010-02-18

In this work, the constrained equilibrium principle is introduced and applied to the derivations of the nonequilibrium solvation free energy and solvent reorganization energy in the process of removing the hydrated electron. Within the framework of the continuum model, a modified expression of the vertical detachment energy (VDE) of a hydrated electron in water is formulated. Making use of the approximation of spherical cavity and point charge, the variation tendency of VDE accompanying the size increase of the water cluster has been inspected. Discussions comparing the present form of the VDE and the traditional one and the influence of the cavity radius in either the fixed pattern or the varying pattern on the VDE have been made.

Charge-transfer spectra of tetravalent lanthanide ions in oxides

NARCIS (Netherlands)

Hoefdraad, H.E.

The charge-transfer spectra of Ce4+, Pr4+ and Tb4+ in a number of oxides are reported. It is noted that the position of the first charge-transfer band is fixed for the metal ion in an oxygen coordination of VI, but varies in VIII coordination as a function of the host lattice. It is argued that this

Functional relevance of G-protein-coupled-receptor-associated proteins, exemplified by receptor-activity-modifying proteins (RAMPs).

Science.gov (United States)

Fischer, J A; Muff, R; Born, W

2002-08-01

The calcitonin (CT) receptor (CTR) and the CTR-like receptor (CRLR) are close relatives within the type II family of G-protein-coupled receptors, demonstrating sequence identity of 50%. Unlike the interaction between CT and CTR, receptors for the related hormones and neuropeptides amylin, CT-gene-related peptide (CGRP) and adrenomedullin (AM) require one of three accessory receptor-activity-modifying proteins (RAMPs) for ligand recognition. An amylin/CGRP receptor is revealed when CTR is co-expressed with RAMP1. When complexed with RAMP3, CTR interacts with amylin alone. CRLR, initially classed as an orphan receptor, is a CGRP receptor when co-expressed with RAMP1. The same receptor is specific for AM in the presence of RAMP2. Together with human RAMP3, CRLR defines an AM receptor, and with mouse RAMP3 it is a low-affinity CGRP/AM receptor. CTR-RAMP1, antagonized preferentially by salmon CT-(8-32) and not by CGRP-(8-37), and CRLR-RAMP1, antagonized by CGRP-(8-37), are two CGRP receptor isotypes. Thus amylin and CGRP interact specifically with heterodimeric complexes between CTR and RAMP1 or RAMP3, and CGRP and AM interact with complexes between CRLR and RAMP1, RAMP2 or RAMP3.

Formation of supported lipid bilayers of charged E. coli lipids on modified gold by vesicle fusion

Directory of Open Access Journals (Sweden)

Ileana F. Márquez

2017-01-01

Full Text Available We describe a simple way of fusing E. coli lipid vesicles onto a gold surface. Supported lipid bilayers on metal surfaces are interesting for several reasons: transducing a biological signal to an electric readout, using surface analytical tools such as Surface Plasmon Resonance (SPR, Infrared Reflection Absorption Spectroscopy, Neutron Reflectivity or Electrochemistry. The most widely used method to prepare supported lipid membranes is fusion of preexisting liposomes. It is quite efficient on hydrophilic surfaces such as glass, mica or SiO2, but vesicle fusion on metals and metal oxide surfaces (as gold, titanium oxide or indium tin oxide, remains a challenge, particularly for vesicles containing charged lipids, as is the case of bacterial lipids. We describe a simple method based on modifying the gold surface with a charged mercaptopropionic acid self-assembled monolayer and liposomes partially solubilized with detergent. The formed bilayers were characterized using a Quartz Crystal Microbalance with dissipation (QCM-D and Atomic Force Microscopy (AFM. Some advantages of this protocol are that the stability of the self-assembled monolayer allows for repeated use of the substrate after detergent removal of the bilayer and that the amount of detergent required for optimal fusion can be determined previously using the lipid-detergent solubility curve.

Efficient wireless power charging of electric vehicle by modifying the magnetic characteristics of the medium

Science.gov (United States)

Mahmud, Mohammad Hazzaz

There is a developing enthusiasm for electric vehicle (EV) innovations as a result of their lessened fuel utilization and greenhouse emission especially through wireless power transfer (WPT) due to the convenience and continuous charging. Numerous research initiatives target on wireless power transfer (WPT) system in the attempt to improve the transportation for last few decades. But several problems like less efficiency, high frequency, long distance energy transfer etc. were always been occupied by the wireless power transfer system. Two ideas have been developed in this research to resolve the two main problems of WPT for electric vehicles which are low efficiency due to large distance between the two coils and slow charging time. As the first phase of study, a proper model, including the coils and cores were required. The selected model was a finite element (FE) modeling. Another part of this study was to create a modified cement that will act as a semi-conductive material for covering the transmitting antenna area. A high frequency wide band gap switch will be used for transferring high amount of power in a very short time. More over this research also proves that, if cores could be added with the transmitter coil and receiver coil then the output efficiency dramatically increased comparing with without core model of transmitter and receiver. The wireless charging is not restricted to parking lot, since it's planned to be embedded into parking space concrete or roadway concrete or asphalt. Therefore, it can also be installed at junctions (behind red lights), stop signs or any spot that the vehicle might stop for several moments. This technology will become more feasible, if the charging time decreases. Therefore, a new model of for wireless power transfer has been proposed in this study which has shown significant improvement. Another motive of this study was to improve the conductivity and permeability in such a way that the medium that is on the top of the

Profiling an electrospray plume by laser-induced fluorescence and Fraunhofer diffraction combined to mass spectrometry: influence of size and composition of droplets on charge-state distributions of electrosprayed proteins.

Science.gov (United States)

Girod, Marion; Dagany, Xavier; Boutou, Véronique; Broyer, Michel; Antoine, Rodolphe; Dugourd, Philippe; Mordehai, Alex; Love, Craig; Werlich, Mark; Fjeldsted, John; Stafford, George

2012-07-14

We investigated how physico-chemical properties of charged droplets are affected by the electrospray process, using simultaneous in situ measurements by laser-induced fluorescence (LIF), Fraunhofer diffraction and mass spectrometry. For this purpose, we implemented a laser-induced-fluorescence profiling setup in conjunction with a fast, high-resolution particle sizing scheme on a modified Agilent Jet Stream electrospray source coupled to a single quadrupole mass analyser. The optical setup permits us to profile the solvent fractionation and the size of the droplets as they evaporate in an electrospray plume by measuring both the angular scattering pattern and emission spectra of a solvatochromic fluorescent dye. Mass spectra are recorded simultaneously. These mass spectrometry and optical spectroscopy investigations allow us to study the relation between the observed charge-state distributions of protein anions and physico-chemical properties of evaporating droplets in the spray plume. By mixing water with methanol, a refolding of cytochrome C is observed as the water percentage increases in the plume due to the preponderant evaporation of volatile methanol.

Colloid, adhesive and release properties of nanoparticular ternary complexes between cationic and anionic polysaccharides and basic proteins like bone morphogenetic protein BMP-2.

Science.gov (United States)

Petzold, R; Vehlow, D; Urban, B; Grab, A L; Cavalcanti-Adam, E A; Alt, V; Müller, M

2017-03-01

Herein we describe an interfacial local drug delivery system for bone morphogenetic protein 2 (BMP-2) based on coatings of polyelectrolyte complex (PEC) nanoparticles (NP). The application horizon is the functionalization of bone substituting materials (BSM) used for the therapy of systemic bone diseases. Nanoparticular ternary complexes of cationic and anionic polysaccharides and BMP-2 or two further model proteins, respectively, were prepared in dependence of the molar mixing ratio, pH value and of the cationic polysaccharide. As further proteins chymotrypsin (CHY) and papain (PAP) were selected, which served as model proteins for BMP-2 due to similar isoelectric points and molecular weights. As charged polysaccharides ethylenediamine modified cellulose (EDAC) and trimethylammonium modified cellulose (PQ10) were combined with cellulose sulphatesulfate (CS). Mixing diluted cationic and anionic polysaccharide and protein solutions according to a slight either anionic or cationic excess charge colloidal ternary dispersions formed, which were cast onto germanium model substrates by water evaporation. Dynamic light scattering (DLS) demonstrated, that these dispersions were colloidally stable for at least one week. Fourier Transform Infrared (FTIR) showed, that the cast protein loaded PEC NP coatings were irreversibly adhesive at the model substrate in contact to HEPES buffer and solely CHY, PAP and BMP-2 were released within long-term time scale. Advantageously, out of the three proteins BMP-2 showed the smallest initial burst and the slowest release kinetics and around 25% of the initial BMP-2 content were released within 14days. Released BMP-2 showed significant activity in the myoblast cells indicating the ability to regulate the formation of new bone. Therefore, BMP-2 loaded PEC NP are suggested as novel promising tool for the functionalization of BSM used for the therapy of systemic bone diseases. Copyright © 2016 Elsevier B.V. All rights reserved.

Positron-electron annihilation in a field of a fixed electric dipole

International Nuclear Information System (INIS)

Butkevich, E.R.; Sabirov, R.Kh.

1989-01-01

Positron annihilation from dipole Ps state conditioned by simultaneous localization of positron and electron in the field of fixed electric dipole is considered. Calculation of annihilation γ-quantum angular correlation curve is performed. For the fixed value of the dipole moment the correlation curve half-width is reduced with the reduction of dipole charges. Correlation curve narrowing takes place with the growth of dipole moment. It is stressed that when interpreting the narrow component one should be careful when choosing the appropriate annihilation mechanism. Experimental result analysis is given

Optimized Mitochondrial Targeting of Proteins Encoded by Modified mRNAs Rescues Cells Harboring Mutations in mtATP6

Directory of Open Access Journals (Sweden)

Randall Marcelo Chin

2018-03-01

Full Text Available Summary: Mitochondrial disease may be caused by mutations in the protein-coding genes of the mitochondrial genome. A promising strategy for treating such diseases is allotopic expression—the translation of wild-type copies of these proteins in the cytosol, with subsequent translocation into the mitochondria, resulting in rescue of mitochondrial function. In this paper, we develop an automated, quantitative, and unbiased screening platform to evaluate protein localization and mitochondrial morphology. This platform was used to compare 31 mitochondrial targeting sequences and 15 3′ UTRs in their ability to localize up to 9 allotopically expressed proteins to the mitochondria and their subsequent impact on mitochondrial morphology. Taking these two factors together, we synthesized chemically modified mRNAs that encode for an optimized allotopic expression construct for mtATP6. These mRNAs were able to functionally rescue a cell line harboring the 8993T > G point mutation in the mtATP6 gene. : Allotopic expression of proteins normally encoded by mtDNA is a promising therapy for mitochondrial disease. Chin et al. use an unbiased and high-content imaging-based screening platform to optimize allotopic expression. Modified mRNAs encoding for the optimized allotopic expression constructs rescued the respiration and growth of mtATP6-deficient cells. Keywords: mitochondria, mitochondrial disease, mRNA, modified mRNA, ATP6, allotopic expression, rare disease, gene therapy, screening, high content imaging

Introduction to charm decay analysis in fixed target experiments

Energy Technology Data Exchange (ETDEWEB)

Bediaga, Ignacio; Goebel, Carla

1996-01-01

We present an introduction to data analysis in Experimental High Energy Physics, and some concepts and useful tools are discussed. To illustrate, we use the data of E-791, a fixed target experiment recently realized at Fermilab. In particular, we analyse decay modes of D{sup +} meson with three charged particles in the final state. (author). 8 refs., 22 figs., 1 tab.

Introduction to charm decay analysis in fixed target experiments

International Nuclear Information System (INIS)

Bediaga, Ignacio; Goebel, Carla.

1996-01-01

We present an introduction to data analysis in Experimental High Energy Physics, and some concepts and useful tools are discussed. To illustrate, we use the data of E-791, a fixed target experiment recently realized at Fermilab. In particular, we analyse decay modes of D + meson with three charged particles in the final state. (author). 8 refs., 22 figs., 1 tab

Protein sensing by nanofluidic crystal and its signal enhancement

Science.gov (United States)

Sang, Jianming; Du, Hongtan; Wang, Wei; Chu, Ming; Wang, Yuedan; Li, Haichao; Alice Zhang, Haixia; Wu, Wengang; Li, Zhihong

2013-01-01

Nanofluidics has a unique property that ionic conductance across a nanometer-sized confined space is strongly affected by the space surface charge density, which can be utilized to construct electrical read-out biosensor. Based on this principle, this work demonstrated a novel protein sensor along with a sandwich signal enhancement approach. Nanoparticles with designed aptamer onside are assembled in a suspended micropore to form a 3-dimensional network of nanometer-sized interstices, named as nanofluidic crystal hereafter, as the basic sensing unit. Proteins captured by aptamers will change the surface charge density of nanoparticles and thereby can be detected by monitoring the ionic conductance across this nanofluidic crystal. Another aptamer can further enlarge the variations of the surface charge density by forming a sandwich structure (capturing aptamer/protein/signal enhancement aptamer) and the read-out conductance as well. The preliminary experimental results indicated that human α-thrombin was successfully detected by the corresponding aptamer modified nanofluidic crystal with the limit of detection of 5 nM (0.18 μg/ml) and the read-out signal was enhanced up to 3 folds by using another thrombin aptamer. Being easy to graft probe, facile and low-cost to prepare the nano-device, and having an electrical read-out, the present nanofluidic crystal scheme is a promising and universal strategy for protein sensing. PMID:24404017

Gauge-fixing parameter dependence of two-point gauge-variant correlation functions

International Nuclear Information System (INIS)

Zhai, C.

1996-01-01

The gauge-fixing parameter ξ dependence of two-point gauge-variant correlation functions is studied for QED and QCD. We show that, in three Euclidean dimensions, or for four-dimensional thermal gauge theories, the usual procedure of getting a general covariant gauge-fixing term by averaging over a class of covariant gauge-fixing conditions leads to a nontrivial gauge-fixing parameter dependence in gauge-variant two-point correlation functions (e.g., fermion propagators). This nontrivial gauge-fixing parameter dependence modifies the large-distance behavior of the two-point correlation functions by introducing additional exponentially decaying factors. These factors are the origin of the gauge dependence encountered in some perturbative evaluations of the damping rates and the static chromoelectric screening length in a general covariant gauge. To avoid this modification of the long-distance behavior introduced by performing the average over a class of covariant gauge-fixing conditions, one can either choose a vanishing gauge-fixing parameter or apply an unphysical infrared cutoff. copyright 1996 The American Physical Society

Modifying a standard method allows simultaneous extraction of RNA and protein, enabling detection of enzymes in the rat retina with low expressions and protein levels.

Science.gov (United States)

Agardh, Elisabet; Gustavsson, Carin; Hagert, Per; Nilsson, Marie; Agardh, Carl-David

2006-02-01

The aim of the study was to evaluate messenger RNA and protein expression in limited amounts of tissue with low protein content. The Chomczynski method was used for simultaneous extraction of RNA, and protein was modified in the protein isolation step. Template mass and cycling time for the complementary DNA synthesis step of real-time reverse transcription-polymerase chain reaction (RT-PCR) for analysis of catalase, copper/zinc superoxide dismutase, manganese superoxide dismutase, the catalytic subunit of glutamylcysteine ligase, glutathione peroxidase 1, and the endogenous control cyclophilin B (CypB) were optimized before PCR. Polymerase chain reaction accuracy and efficacy were demonstrated by calculating the regression (R2) values of the separate amplification curves. Appropriate antibodies, blocking buffers, and running conditions were established for Western blot, and protein detection and multiplex assays with CypB were performed for each target. During the extraction procedure, the protein phase was dissolved in a modified washing buffer containing 0.1% sodium dodecyl sulfate, followed by ultrafiltration. Enzyme expression on real-time RT-PCR was accomplished with high reliability and reproducibility (R2, 0.990-0.999), and all enzymes except for glutathione peroxidase 1 were detectable in individual retinas on Western blot. Western blot multiplexing with CypB was possible for all targets. In conclusion, connecting gene expression directly to protein levels in the individual rat retina was possible by simultaneous extraction of RNA and protein. Real-time RT-PCR and Western blot allowed accurate detection of retinal protein expressions and levels.

Modelling and Fixed Step Simulation of a Turbo Charged Diesel Engine

OpenAIRE

Ritzén, Jesper

2003-01-01

Having an engine model that is accurate but not too complicated is desirable when working with on-board diagnosis or engine control. In this thesis a four state mean value model is introduced. To make the model usable in an on-line automotive application it is discrete and simulated with a fixed step size solver. Modelling is done with simplicity as main object. Some simple static models are also presented. To validate the model measuring is carried out in a Scania R124LB truck with a 12 lit...

A modified space charge routine for high intensity bunched beams

International Nuclear Information System (INIS)

Lapostolle, P.; Lombardi, A.M.; Tanke, E.; Valero, S.; Garnett, R.W.; Wangler, T.P.

1996-01-01

A new routine and a computer code (DYNAC) for the calculation of space charge densities in a new generation of linear accelerators for various industrial applications is presented. The new beam dynamics method used in this code, employs a set of quasi-Liouvillian equations, allowing beam dynamics computations in long and complex structures for electrons, as well as protons and ions. With this new beam dynamics method, the coordinates of particles are known at any position in the accelerating elements, allowing multistep space charge calculations. (K.A.)

How Chain Length and Charge Affect Surfactant Denaturation of Acyl Coenzyme A Binding Protein (ACBP)

DEFF Research Database (Denmark)

Andersen, Kell Kleiner; Otzen, Daniel

2009-01-01

maltoside (DDM). The aim has been to determine how surfactant chain length and micellar charge affect the denaturation mechanism. ACBP denatures in two steps irrespective of surfactant chain length, but with increasing chain length, the potency of the denaturant rises more rapidly than the critical micelle......Using intrinsic tryptophan fluorescence, equilibria and kinetics of unfolding of acyl coenzyme A binding protein (ACBP) have been investigated in sodium alkyl sulfate surfactants of different chain length (8-16 carbon atoms) and with different proportions of the nonionic surfactant dodecyl...... constants increases linearly with denaturant concentration below the cmc but declines at higher concentrations. Both shortening chain length and decreasing micellar charge reduce the overall kinetics of unfolding and makes the dependence of unfolding rate constants on surfactant concentration more complex...

How to Estimate Demand Charge Savings from PV on Commercial Buildings

Energy Technology Data Exchange (ETDEWEB)

Gagnon, Pieter J [National Renewable Energy Laboratory (NREL), Golden, CO (United States); Bird, Lori A [National Renewable Energy Laboratory (NREL), Golden, CO (United States)

2017-09-28

Rooftop photovoltaic (PV) systems are compensated through retail electricity tariffs - and for commercial and industrial customers, these are typically comprised of three components: a fixed monthly charge, energy charges, and demand charges. Of these, PV's ability to reduce demand charges has traditionally been the most difficult to estimate. In this fact sheet we explain the basics of demand charges, and provide a new method that a potential customer or PV developer can use to estimate a range of potential demand charge savings for a proposed PV system. These savings can then be added to other project cash flows, in assessing the project's financial performance.

New Fixed-Target Experiments to Search for Dark Gauge Forces

Energy Technology Data Exchange (ETDEWEB)

Bjorken, James D.; Essig, Rouven; Schuster, Philip; /SLAC; Toro, Natalia; /Stanford U., ITP

2010-06-11

Fixed-target experiments are ideally suited for discovering new MeV-GeV mass U(1) gauge bosons through their kinetic mixing with the photon. In this paper, we identify the production and decay properties of new light gauge bosons that dictate fixed-target search strategies. We summarize existing limits and suggest five new experimental approaches that we anticipate can cover most of the natural parameter space, using currently operating GeV-energy beams and well-established detection methods. Such experiments are particularly timely in light of recent terrestrial and astrophysical anomalies (PAMELA, FERMI, DAMA/LIBRA, etc.) consistent with dark matter charged under a new gauge force.

The dipole moment of a wall-charged void in a bulk dielectric

DEFF Research Database (Denmark)

McAllister, Iain Wilson

1993-01-01

The dipole moment of a wall-charged void is examined with reference to the spatial extent of the surface charge density σ and the distribution of this charge. The salient factors influencing the void dipole moment are also examined. From a study of spherical voids, it is shown that, although the σ......-distribution influences the dipole moment, the spatial extent of σ has a greater influence. This behavior is not unexpected. For a void of fixed dimensions, the smaller the charged surface area, the greater is the charges, and thus the greater the dipole moment...

Payload charging events in the mesosphere and their impact on Langmuir type electric probes

Directory of Open Access Journals (Sweden)

T. A. Bekkeng

2013-02-01

Full Text Available Three sounding rockets were launched from Andøya Rocket Range in the ECOMA campaign in December 2010. The aim was to study the evolution of meteoric smoke particles during a major meteor shower. Of the various instruments onboard the rocket payload, this paper presents the data from a multi-Needle Langmuir Probe (m-NLP and a charged dust detector. The payload floating potential, as observed using the m-NLP instrument, shows charging events on two of the three flights. These charging events cannot be explained using a simple charging model, and have implications towards the use of fixed bias Langmuir probes on sounding rockets investigating mesospheric altitudes. We show that for a reliable use of a single fixed bias Langmuir probe as a high spatial resolution relative density measurement, each payload should also carry an additional instrument to measure payload floating potential, and an instrument that is immune to spacecraft charging and measures absolute plasma density.

X-ray crystallographic analysis of adipocyte fatty acid binding protein (aP2) modified with 4-hydroxy-2-nonenal

Energy Technology Data Exchange (ETDEWEB)

Hellberg, Kristina; Grimsrud, Paul A.; Kruse, Andrew C.; Banaszak, Leonard J.; Ohlendorf, Douglas H.; Bernlohr, David A. (UMM)

2012-07-11

Fatty acid binding proteins (FABP) have been characterized as facilitating the intracellular solubilization and transport of long-chain fatty acyl carboxylates via noncovalent interactions. More recent work has shown that the adipocyte FABP is also covalently modified in vivo on Cys117 with 4-hydroxy-2-nonenal (4-HNE), a bioactive aldehyde linked to oxidative stress and inflammation. To evaluate 4-HNE binding and modification, the crystal structures of adipocyte FABP covalently and noncovalently bound to 4-HNE have been solved to 1.9 {angstrom} and 2.3 {angstrom} resolution, respectively. While the 4-HNE in the noncovalently modified protein is coordinated similarly to a carboxylate of a fatty acid, the covalent form show a novel coordination through a water molecule at the polar end of the lipid. Other defining features between the two structures with 4-HNE and previously solved structures of the protein include a peptide flip between residues Ala36 and Lys37 and the rotation of the side chain of Phe57 into its closed conformation. Representing the first structure of an endogenous target protein covalently modified by 4-HNE, these results define a new class of in vivo ligands for FABPs and extend their physiological substrates to include bioactive aldehydes.

Cl II Malocclusion Treatment, Using the Modified Twin Block Appliance Coordinated with Fixed Orthodontics in a Postmenarche Patient

Directory of Open Access Journals (Sweden)

Amin Aminian

2017-01-01

Full Text Available Functional appliances have been used for treatment of Class II patients for a long time. The main objective of therapy with functional appliances is to induce supplementary lengthening of the mandible by stimulating increased growth at the condylar cartilage. The Twin Block appliance is one of the most commonly used functional appliances. The aim of this paper is to present a case report of mandibular deficiency treatment with Twin Block appliance in a female patient whose sexual maturation (one and a half years after menarche and cervical vertebral maturation stage indicated the end of the growth peak. The treatment started with bonding 0.022 in MBT prescription brackets on the upper arch in order to align upper teeth and create a symmetric overjet. When reaching alignment, a modified Twin Block was given to the patient for 8 months. Final coordination was achieved with fixed appliances in both arches. At the end of the treatment, profile of the patient improved, crowding was relieved, and Cl I relationship with normal overjet and overbite was achieved.

Noise power spectrum of the fixed pattern noise in digital radiography detectors

Energy Technology Data Exchange (ETDEWEB)

Kim, Dong Sik, E-mail: dskim@hufs.ac.kr [Department of Electronics Engineering, Hankuk University of Foreign Studies, Gyeonggi-do 449-791 (Korea, Republic of); Kim, Eun [R& D Center, DRTECH Co., Gyeonggi-do 13558 (Korea, Republic of)

2016-06-15

Purpose: The fixed pattern noise in radiography image detectors is caused by various sources. Multiple readout circuits with gate drivers and charge amplifiers are used to efficiently acquire the pixel voltage signals. However, the multiple circuits are not identical and thus yield nonuniform system gains. Nonuniform sensitivities are also produced from local variations in the charge collection elements. Furthermore, in phosphor-based detectors, the optical scattering at the top surface of the columnar CsI growth, the grain boundaries, and the disorder structure causes spatial sensitivity variations. These nonuniform gains or sensitivities cause fixed pattern noise and degrade the detector performance, even though the noise problem can be partially alleviated by using gain correction techniques. Hence, in order to develop good detectors, comparative analysis of the energy spectrum of the fixed pattern noise is important. Methods: In order to observe the energy spectrum of the fixed pattern noise, a normalized noise power spectrum (NNPS) of the fixed pattern noise is considered in this paper. Since the fixed pattern noise is mainly caused by the nonuniform gains, we call the spectrum the gain NNPS. We first asymptotically observe the gain NNPS and then formulate two relationships to calculate the gain NNPS based on a nonuniform-gain model. Since the gain NNPS values are quite low compared to the usual NNPS, measuring such a low NNPS value is difficult. By using the average of the uniform exposure images, a robust measuring method for the gain NNPS is proposed in this paper. Results: By using the proposed measuring method, the gain NNPS curves of several prototypes of general radiography and mammography detectors were measured to analyze their fixed pattern noise properties. We notice that a direct detector, which is based on the a-Se photoconductor, showed lower gain NNPS than the indirect-detector case, which is based on the CsI scintillator. By comparing the gain

Introducing Membrane Charge and Membrane Potential to T Cell Signaling

Directory of Open Access Journals (Sweden)

Yuanqing Ma

2017-11-01

Full Text Available While membrane models now include the heterogeneous distribution of lipids, the impact of membrane charges on regulating the association of proteins with the plasma membrane is often overlooked. Charged lipids are asymmetrically distributed between the two leaflets of the plasma membrane, resulting in the inner leaflet being negatively charged and a surface potential that attracts and binds positively charged ions, proteins, and peptide motifs. These interactions not only create a transmembrane potential but they can also facilitate the formation of charged membrane domains. Here, we reference fields outside of immunology in which consequences of membrane charge are better characterized to highlight important mechanisms. We then focus on T cell receptor (TCR signaling, reviewing the evidence that membrane charges and membrane-associated calcium regulate phosphorylation of the TCR–CD3 complex and discuss how the immunological synapse exhibits distinct patterns of membrane charge distribution. We propose that charged lipids, ions in solution, and transient protein interactions form a dynamic equilibrium during T cell activation.

Integration of carboxyl modified magnetic particles and aqueous two-phase extraction for selective separation of proteins.

Science.gov (United States)

Gai, Qingqing; Qu, Feng; Zhang, Tao; Zhang, Yukui

2011-07-15

Both of the magnetic particle adsorption and aqueous two-phase extraction (ATPE) were simple, fast and low-cost method for protein separation. Selective proteins adsorption by carboxyl modified magnetic particles was investigated according to protein isoelectric point, solution pH and ionic strength. Aqueous two-phase system of PEG/sulphate exhibited selective separation and extraction for proteins before and after magnetic adsorption. The two combination ways, magnetic adsorption followed by ATPE and ATPE followed by magnetic adsorption, for the separation of proteins mixture of lysozyme, bovine serum albumin, trypsin, cytochrome C and myloglobin were discussed and compared. The way of magnetic adsorption followed by ATPE was also applied to human serum separation. Copyright © 2011 Elsevier B.V. All rights reserved.

Electrostatic charge characteristics of jet nebulized aerosols.

Science.gov (United States)

Kwok, Philip Chi Lip; Trietsch, Sebastiaan J; Kumon, Michiko; Chan, Hak-Kim

2010-06-01

Liquid droplets can be spontaneously charged in the absence of applied electric fields by spraying. It has been shown by computational simulation that charges may influence particle deposition in the airways. The electrostatic properties of jet nebulized aerosols and their potential effects on lung deposition have hardly been studied. A modified electrical low pressure impactor (ELPI) was employed to characterize the aerosol charges generated from jet nebulized commercial products. The charge and size measurements were conducted at 50% RH and 22 degrees C with a modified ELPI. Ventolin, Bricanyl, and Atrovent were nebulized using PARI LC Plus jet nebulizers coupled to a DeVilbiss Pulmo-Aide compressor. The aerosols were sampled in 30-sec durations. The drug deposits on the impactor stages were assayed chemically using high-performance liquid chromatography (HPLC). The charges of nebulized deionized water, isotonic saline, and the three commercial products diluted with saline were also measured to analyze the contributions of the major nebule ingredients on charging. No mass assays were performed on these runs. All three commercial nebules generated net negative charges. The magnitude of the charges reduced over the period of nebulization. Ventolin and Bricanyl yielded similar charge profiles. Highly variable charges were produced from deionized water. On the other hand, nebulized saline reproducibly generated net positive charges. Diluted commercial nebules showed charge polarity inversion. The charge profiles of diluted salbutamol and terbutaline solutions resembled those of saline, while the charges from diluted ipratropium solutions fluctuated near neutrality. The charge profiles were shown to be influenced by the concentration and physicochemical properties of the drugs, as well as the history of nebulization. The drugs may have unique isoelectric concentrations in saline at which the nebulized droplets would carry near-zero charges. According to results from

On the charge radius of the neutrino

CERN Document Server

Bernabeu, J; Papavassiliou, J; Vidal, J

2000-01-01

Using the pinch technique we construct at one-loop order a neutrino charge radius, which is finite, depends neither on the gauge-fixing parameter nor on the gauge-fixing scheme employed, and is process-independent. This definition stems solely from an effective proper photon-neutrino one-loop vertex, with no reference to box or self-energy contributions. The role of the $WW$ box in this construction is critically examined. In particular it is shown that the exclusion of the effective WW box from the definition of the neutrino charge radius is not a matter of convention but is in fact dynamically realized when the target-fermions are right-handedly polarized. In this way we obtain a unique decomposition of effective self-energies, vertices, and boxes, which separately respect electroweak gauge invariance. We elaborate on the tree-level origin of the mechanism which enforces at one-loop level massive cancellations among the longitudinal momenta appearing in the Feynman diagrams, and in particular those associat...

The Tendencies of Piece Casting from Modified Irons

Directory of Open Access Journals (Sweden)

Cinca Ionel Lupinca

2010-10-01

Full Text Available In this paper we have presented the metalographic studies made on the grey cast irons treated with complex modifying substances, type FeSiMgRE (Mg alloy and their influence on the compactness degree of graphite separations. For research and experiments, a melt of grey iron was produced in an induction furnace of a capacity of 5to, starting with a metallic charge made from 100% synthetic pig iron. We realized eight practical charge made modification, by using different combinations of modifying substance and in different concentrations. The addition of carbon to the melt was performed using electrode graphite powder in the metallic charge.

Experimental determination of net protein charge and A(tot) and K(a) of nonvolatile buffers in canine plasma.

Science.gov (United States)

Constable, Peter D; Stämpfli, Henry R

2005-01-01

Acid-base abnormalities frequently are present in sick dogs. The mechanism for an acid-base disturbance can be determined with the simplified strong ion approach, which requires accurate values for the total concentration of plasma nonvolatile buffers (A(tot)) and the effective dissociation constant for plasma weak acids (K(a)). The aims of this study were to experimentally determine A(tot) and K(a) values for canine plasma. Plasma was harvested from 10 healthy dogs; the concentrations of quantitatively important strong ions (Na+, K+, Ca2+, Mg2+, Cl-, L-lactate) and nonvolatile buffer ions (total protein, albumin, phosphate) were determined; and the plasma was tonometered with CO2 at 37 degrees C. Strong ion difference (SID) was calculated from the measured strong ion concentrations, and nonlinear regression was used to estimate values for A(tot) and K(a), which were validated with data from an in vitro and in vivo study. Mean (+/- SD) values for canine plasma were A(tot) = (17.4 +/- 8.6) mM (equivalent to 0.273 mmol/g of total protein or 0.469 mmol/g of albumin); K(a) = (0.17 +/- 0.11) x 10(-7); pK(a) = 7.77. The calculated SID for normal canine plasma (pH = 7.40; P(CO2) = 37 mm Hg; [total protein] = 64 g/L) was 27 mEq/L. The net protein charge for normal canine plasma was 0.25 mEq/g of total protein or 0.42 mEq/g of albumin. Application of the experimentally determined values for A(tot), K(a), and net protein charge should improve understanding of the mechanism for complex acid-base disturbances in dogs.

Charge-regularized swelling kinetics of polyelectrolyte gels: Elasticity and diffusion

Science.gov (United States)

Sen, Swati; Kundagrami, Arindam

2017-11-01

We apply a recently developed method [S. Sen and A. Kundagrami, J. Chem. Phys. 143, 224904 (2015)], using a phenomenological expression of osmotic stress, as a function of polymer and charge densities, hydrophobicity, and network elasticity for the swelling of spherical polyelectrolyte (PE) gels with fixed and variable charges in a salt-free solvent. This expression of stress is used in the equation of motion of swelling kinetics of spherical PE gels to numerically calculate the spatial profiles for the polymer and free ion densities at different time steps and the time evolution of the size of the gel. We compare the profiles of the same variables obtained from the classical linear theory of elasticity and quantitatively estimate the bulk modulus of the PE gel. Further, we obtain an analytical expression of the elastic modulus from the linearized expression of stress (in the small deformation limit). We find that the estimated bulk modulus of the PE gel decreases with the increase of its effective charge for a fixed degree of deformation during swelling. Finally, we match the gel-front locations with the experimental data, taken from the measurements of charged reversible addition-fragmentation chain transfer gels to show an increase in gel-size with charge and also match the same for PNIPAM (uncharged) and imidazolium-based (charged) minigels, which specifically confirms the decrease of the gel modulus value with the increase of the charge. The agreement between experimental and theoretical results confirms general diffusive behaviour for swelling of PE gels with a decreasing bulk modulus with increasing degree of ionization (charge). The new formalism captures large deformations as well with a significant variation of charge content of the gel. It is found that PE gels with large deformation but same initial size swell faster with a higher charge.

Physical Stability of Octenyl Succinate-Modified Polysaccharides and Whey Proteins for Potential Use as Bioactive Carriers in Food Systems.

Science.gov (United States)

Puerta-Gomez, Alex F; Castell-Perez, M Elena

2015-06-01

The high cost and potential toxicity of biodegradable polymers like poly(lactic-co-glycolic)acid (PLGA) has increased the interest in natural and modified biopolymers as bioactive carriers. This study characterized the physical stability (water sorption and state transition behavior) of selected starch and proteins: octenyl succinate-modified depolymerized waxy corn starch (DWxCn), waxy rice starch (DWxRc), phytoglycogen, whey protein concentrate (80%, WPC), whey protein isolate (WPI), and α-lactalbumin (α-L) to determine their potential as carriers of bioactive compounds under different environmental conditions. After enzyme modification and particle size characterization, glass transition temperature and moisture isotherms were used to characterize the systems. DWxCn and DWxRc had increased water sorption compared to native starch. The level of octenyl succinate anhydrate (OSA) modification (3% and 7%) did not reduce the water sorption of the DWxCn and phytoglycogen samples. The Guggenheim-Andersen-de Boer model indicated that native waxy corn had significantly (P whey proteins had higher glass transition temperature (Tg) values. On the other hand, depolymerized waxy starches at 7%-OSA modification had a "melted" appearance when exposed to environments with high relative humidity (above 70%) after 10 days at 23 °C. The use of depolymerized and OSA-modified polysaccharides blended with proteins created more stable blends of biopolymers. Hence, this biopolymer would be suitable for materials exposed to high humidity environments in food applications. © 2015 Institute of Food Technologists®

Enhanced charge efficiency and reduced energy use in capacitive deionization by increasing the discharge voltage.

Science.gov (United States)

Kim, T; Dykstra, J E; Porada, S; van der Wal, A; Yoon, J; Biesheuvel, P M

2015-05-15

Capacitive deionization (CDI) is an electrochemical method for water desalination using porous carbon electrodes. A key parameter in CDI is the charge efficiency, Λ, which is the ratio of salt adsorption over charge in a CDI-cycle. Values for Λ in CDI are typically around 0.5-0.8, significantly less than the theoretical maximum of unity, due to the fact that not only counterions are adsorbed into the pores of the carbon electrodes, but at the same time coions are released. To enhance Λ, ion-exchange membranes (IEMs) can be implemented. With membranes, Λ can be close to unity because the membranes only allow passage for the counterions. Enhancing the value of Λ is advantageous as this implies a lower electrical current and (at a fixed charging voltage) a reduced energy use. We demonstrate how, without the need to include IEMs, the charge efficiency can be increased to values close to the theoretical maximum of unity, by increasing the cell voltage during discharge, with only a small loss of salt adsorption capacity per cycle. In separate constant-current CDI experiments, where after some time the effluent salt concentration reaches a stable value, this value is reached earlier with increased discharge voltage. We compare the experimental results with predictions of porous electrode theory which includes an equilibrium Donnan electrical double layer model for salt adsorption in carbon micropores. Our results highlight the potential of modified operational schemes in CDI to increase charge efficiency and reduce energy use of water desalination. Copyright © 2014 Elsevier Inc. All rights reserved.

Clinical physics for charged particle treatment planning

International Nuclear Information System (INIS)

Chen, G.T.Y.; Pitluck, S.; Lyman, J.T.

1981-01-01

The installation of a computerized tomography (CT) scanner which can be used with the patient in an upright position is described. This technique will enhance precise location of tumor position relative to critical structures for accurate charged particle dose delivery during fixed horizontal beam radiotherapy. Pixel-by-pixel treatment planning programs have been developed to calculate the dose distribution from multi-port charged particle beams. The plan includes CT scans, data interpretation, and dose calculations. The treatment planning computer is discussed. Treatment planning for irradiation of ocular melanomas is described

Impact of interface charge on the electrostatics of field-plate assisted RESURF devices

NARCIS (Netherlands)

Boksteen, B.K.; Ferrara, A.; Heringa, A.; Steeneken, P.G.; Hueting, Raymond Josephus Engelbart

2014-01-01

A systematic study on the effects of arbitrary parasitic charge profiles, such as trapped or fixed charge, on the 2-D potential distribution in the drain extension of reverse-biased field-plate-assisted reduced surface field (RESURF) devices is presented. Using TCAD device simulations and analytical

TaBoo SeArch Algorithm with a Modified Inverse Histogram for Reproducing Biologically Relevant Rare Events of Proteins.

Science.gov (United States)

Harada, Ryuhei; Takano, Yu; Shigeta, Yasuteru

2016-05-10

The TaBoo SeArch (TBSA) algorithm [ Harada et al. J. Comput. Chem. 2015 , 36 , 763 - 772 and Harada et al. Chem. Phys. Lett. 2015 , 630 , 68 - 75 ] was recently proposed as an enhanced conformational sampling method for reproducing biologically relevant rare events of a given protein. In TBSA, an inverse histogram of the original distribution, mapped onto a set of reaction coordinates, is constructed from trajectories obtained by multiple short-time molecular dynamics (MD) simulations. Rarely occurring states of a given protein are statistically selected as new initial states based on the inverse histogram, and resampling is performed by restarting the MD simulations from the new initial states to promote the conformational transition. In this process, the definition of the inverse histogram, which characterizes the rarely occurring states, is crucial for the efficiency of TBSA. In this study, we propose a simple modification of the inverse histogram to further accelerate the convergence of TBSA. As demonstrations of the modified TBSA, we applied it to (a) hydrogen bonding rearrangements of Met-enkephalin, (b) large-amplitude domain motions of Glutamine-Binding Protein, and (c) folding processes of the B domain of Staphylococcus aureus Protein A. All demonstrations numerically proved that the modified TBSA reproduced these biologically relevant rare events with nanosecond-order simulation times, although a set of microsecond-order, canonical MD simulations failed to reproduce the rare events, indicating the high efficiency of the modified TBSA.

OPTIONS FOR REPORTING OF DEPRECIATION OF FIXED BIOLOGICAL ASSETS ACCORDING TO IAS 41

Directory of Open Access Journals (Sweden)

Elena Georgieva

2016-06-01

Full Text Available An important element of the accounting policies of enterprises, operating in the agricultural sector, is the organization of accountability of fixed biological assets. With the recent changes in the provisions of International Accounting Standards, effective 01.01.2016, bearer plants were removed from the scope of IAS 41 Agriculture and included within the scope of IAS 16 Property, plant and equipment. The aim of the study is to analyze the impact of changes on the accounting policies of agricultural enterprises, in particular the assessment of fixed biological assets and capabilities for charging depreciation. An important aspect is the synchronization of accounting and tax legislation regarding possibilities for the recognition of depreciation of fixed biological assets.

[The effect of hydrophobic surface properties of protein on its resistance to denaturation by organic solvents (using modified alpha-chymotrypsin as an example].

Science.gov (United States)

Kudriashova, E V; Belova, A B; Vinogradov, A A; Mozhaev, V V

1994-03-01

Catalytic activity of covalently modified alpha-chymotrypsin in water/cosolvent solutions was investigated. The stability of chymotrypsin increases upon modification with hydrophilic reagents, such as glyceraldehyde, pyrometallic and succinic anhydrides, and glucosamine. Correlation was observed between the protein's stability in organic solvents and the degree of hydrophilization of the protein's surface. The protein is the more stable, the higher are the modification degree and the hydrophilicity of the modifying residue. At a certain critical hydrophilization degree of chymotrypsin a limit of stability is achieved. The stabilization effect can be accounted for by the fact that the interaction between water molecules on the surface and protein's functional groups become stronger in the hydrophilized protein.

Ultrathin Graphene-Protein Supercapacitors for Miniaturized Bioelectronics.

Science.gov (United States)

Mosa, Islam M; Pattammattel, Ajith; Kadimisetty, Karteek; Pande, Paritosh; El-Kady, Maher F; Bishop, Gregory W; Novak, Marc; Kaner, Richard B; Basu, Ashis K; Kumar, Challa V; Rusling, James F

2017-09-06

Nearly all implantable bioelectronics are powered by bulky batteries which limit device miniaturization and lifespan. Moreover, batteries contain toxic materials and electrolytes that can be dangerous if leakage occurs. Herein, an approach to fabricate implantable protein-based bioelectrochemical capacitors (bECs) employing new nanocomposite heterostructures in which 2D reduced graphene oxide sheets are interlayered with chemically modified mammalian proteins, while utilizing biological fluids as electrolytes is described. This protein-modified reduced graphene oxide nanocomposite material shows no toxicity to mouse embryo fibroblasts and COS-7 cell cultures at a high concentration of 1600 μg mL -1 which is 160 times higher than those used in bECs, unlike the unmodified graphene oxide which caused toxic cell damage even at low doses of 10 μg mL -1 . The bEC devices are 1 μm thick, fully flexible, and have high energy density comparable to that of lithium thin film batteries. COS-7 cell culture is not affected by long-term exposure to encapsulated bECs over 4 d of continuous charge/discharge cycles. These bECs are unique, protein-based devices, use serum as electrolyte, and have the potential to power a new generation of long-life, miniaturized implantable devices.

Controlling the rejection of protein during membrane filtration by adding selected polyelectrolytes

DEFF Research Database (Denmark)

Pinelo, Manuel; Ferrer Roca, Carme; Meyer, Anne S.

2012-01-01

Electrostatic interactions among the charged groups on proteins and/or between proteins and other solutes significantly affect the aggregation/deposition phenomena that induce fouling and decrease permeate flux during membrane purification of proteins. Such interactions can be turned...... help enhance the performance of membrane filtration for fractionation/purification of a target protein by significantly reducing fouling and modifying rejection/selectivity.......) changing the pH, on the permeate flux and membrane transmission of bovin serum albumina (BSA) through a PVDF membrane. The addition of PS-co-AA to the feed solution resulted in significant increases of the BSA transmission at pH 7.4 as compared to the transmission of a pure BSA solution (1g...

Evaluation of a modified IRMA for anti-D quantitation, using 3H protein A

International Nuclear Information System (INIS)

Dumasia, A.; Gupte, S.

1993-01-01

A modified immunoradiometric assay (IRMA) using tritiated ( 3 H) protein A was developed to estimate anti-D concentration. The main advantages of the assay were longer shelf life of the labelled reagent (more than two years); minimum radiation hazard and; low non specific binding. Levels of anti-D were estimated in 23 Rh (D) immunized women. A good correlation of anti-D concentration (μg/ml) with Rh antibody titre was observed (r=+ 0.89, P 3 H protein A IRMA correlated well with the severity of Rh-HDN. This assay could quantitate anti-D in sera having exclusively IgG 3 subtype. (author). 20 refs., 2 figs., 2 tabs

Physical stage of photosynthesis charge separation

Science.gov (United States)

Yakovlev, A. G.; Shuvalov, V. A.

2016-06-01

An analytical review is given concerning the biophysical aspects of light-driven primary charge separation in photosynthesis reaction centers (RCs) which are special pigment-protein complexes residing in a cell membrane. The primary (physical) stage of charge separation occurs in the pico- and femtosecond ranges and consists of transferring an electron along the active A-branch of pigments. The review presents vast factual material on both the general issues of primary photosynthesis and some more specific topics, including (1) the role of the inactive B-branch of pigments, (2) the effect of the protein environment on the charge separation, and (3) the participation of monomeric bacteriochlorophyll BA in primary electron acceptance. It is shown that the electron transfer and stabilization are strongly influenced by crystallographic water and tyrosine M210 molecules from the nearest environment of BA. A linkage between collective nuclear motions and electron transfer upon charge separation is demonstrated. The nature of the high quantum efficiency of primary charge separation reactions is discussed.

Analysis of potential protein-modifying variants in 9000 endometriosis patients and 150000 controls of European ancestry

DEFF Research Database (Denmark)

Sapkota, Yadav; Vivo, Immaculata De; Steinthorsdottir, Valgerdur

2017-01-01

-modifying variants in endometriosis using exome-array genotyping in 7164 cases and 21005 controls, and a replication set of 1840 cases and 129016 controls of European ancestry. Results in the discovery sample identified significant evidence for association with coding variants in single-variant (rs1801232-CUBN...... sufficient power, our results did not identify any protein-modifying variants (MAF > 0.01) with moderate or large effect sizes in endometriosis, although these variants may exist in non-European populations or in high-risk families. The results suggest continued discovery efforts should focus on genotyping...

Multiplicity distributions of charged hadrons in νp and anti νp charged current interactions

International Nuclear Information System (INIS)

Jones, G.T.; Jones, R.W.L.; Kennedy, B.W.; Morrison, D.R.O.; Mobayyen, M.M.; Wainstein, S.; Borner, H.P.; Myatt, G.; Radojicic, D.; Burke, S.; Aderholz, M.; Hantke, D.; Katz, U.F.; Kern, J.; Schmitz, N.; Wittek, W.

1991-10-01

Using data on νp and anti νp charged current interactions from a bubble chamber experiment with BEBC at CERN, the multiplicity distributions of charged hadrons are investigated. The analysis is based on ∝ 20 000 events with incident ν and ∝ 10 000 events with incident anti ν. The invariant mass W of the total hadronic system ranges from 3 GeV to ∝ 14 GeV. The experimental multiplicity distributions are fitted by the binomial function (for different intervals of W and in different intervals of the rapidity y), by the Levy function and the lognormal function. All three parametrizations give acceptable values for χ 2 /NDF. For fixed W, forward and backward multiplicities are found to be uncorrelated. The normalized moments of the charged multiplicity distributions are measured as a function of W. They show a violation of KNO scaling. (orig.)

Multiplicity distributions of charged hadrons in νp and anti νp charged current interactions

International Nuclear Information System (INIS)

Jones, G.T.; Jones, R.W.L.; Kennedy, B.W.; Morrison, D.R.O.; Mobayyen, M.M.; Wainstein, S.; Aderholz, M.; Hantke, D.; Katz, U.F.; Kern, J.; Schmitz, N.; Wittek, W.; Borner, H.P.; Myatt, G.; Radojicic, D.; Burke, S.

1992-01-01

Using data on νp and anti νp charged current interactions from a bubble chamber experiment with BEBC at CERN, the multiplicity distributions of charged hadrons are investigated. The analysis is based on ∝20 000 events with incident ν and ∝10 000 events with incident anti ν. The invariant mass W of the total hadronic system ranges from 3 GeV to ∝14 GeV. The experimental multiplicity distributions are fitted by the binomial function (for different intervals of W and in different intervals of the rapidity y), by the Levy function and the lognormal function. All three parametrizations give acceptable values for χ 2 /NDF. For fixed W, forward and backward multiplicities are found to be uncorrelated. The normalized moments of the charged multiplicity distributions are measured as a function of W. They show a violation of KNO scaling. (orig.)

Crystallization and molecular-replacement studies of the monoclonal antibody mAbR310 specific for the (R)-HNE-modified protein

International Nuclear Information System (INIS)

Ito, Sohei; Tatsuda, Emi; Ishino, Kousuke; Suzuki, Kenichiro; Sakai, Hiroshi; Uchida, Koji

2006-01-01

Antigen-free Fab fragment of mAbR310, which recognizes (R)-HNE modified protein, has been crystallized. Initial phases have been obtained by molecular replacement. 4-Hydroxy-2-nonenal (HNE), a major racemic product of lipid peroxidation, reacts with histidine to form a stable HNE–histidine Michael addition-type adduct possessing three chiral centres in the cyclic hemiacetal structure. Monoclonal antibodies against HNE-modified protein have been widely used for assessing oxidative stress in vitro and in vivo. Here, the purification, crystallization and preliminary crystallographic analysis of a Fab fragment of novel monoclonal antibody R310 (mAbR310), which recognizes (R)-HNE-modified protein, are reported. The Fab fragment of mAbR310 was obtained by digestion with papain, purified and crystallized. Using hanging-drop vapour-diffusion crystallization techniques, crystals of mAbR310 Fab were obtained. The crystal belongs to the monoclinic space group C2 (unit-cell parameters a = 127.04, b = 65.31, c = 64.29 Å, β = 118.88°) and diffracted X-rays to a resolution of 1.84 Å. The asymmetric unit contains one molecule of mAbR310, with a corresponding crystal volume per protein weight of 2.51 Å 3 Da −1 and a solvent content of 51.0%

Detection of African swine fever virus from formalin fixed and non-fixed tissues by polymerase chain reaction

Directory of Open Access Journals (Sweden)

P. D. Luka

2014-10-01

Full Text Available Aim: Formalin fixing and paraffin embedding of tissue samples is one of the techniques for preserving the structural integrity of cells for a very long time. However, extraction and analysis of genomic material from formalin fixed tissue (FFT remains a challenge despite numerous attempts to develop a more effective method. The success of polymerase chain reaction (PCR depends on the quality of DNA extract. Materials and Methods: Here we assessed the conventional method of DNA extraction from FFT for African swine fever virus (ASFV detection. The modified conventional method gave a higher quality DNA when compared with commercially available DNA extraction kits (QIAamp® DNA Mini Kit, DNeasy® Blood and Tissue Kit, and ZR Genomic DNA™ Tissue MiniPrep. Results: An average A260/A280 DNA purity of 0.86-1.68 and 3.22-5.32 μg DNA/mg for formalin fixed and non-fixed tissues, respectively using a conventional method. In a reproducible and three times repeat PCR, the ASFV DNA expected product size of 278 bp was obtained from the DNA extract of the conventional method but not from the DNA extract of the commercial kits. Conclusion: The present study has demonstrated that the conventional method extracts ASFV genome better than commercial kit. In summary, the commercial kit extraction appeared not suitable to purify ASFV DNA from FFT. We, therefore, recommend that the use of the conventional method be considered for African swine fever DNA extraction from FFT.

Optical patterning of trapped charge in nitrogen-doped diamond

Science.gov (United States)

Jayakumar, Harishankar; Henshaw, Jacob; Dhomkar, Siddharth; Pagliero, Daniela; Laraoui, Abdelghani; Manson, Neil B.; Albu, Remus; Doherty, Marcus W.; Meriles, Carlos A.

2016-08-01

The nitrogen-vacancy (NV) centre in diamond is emerging as a promising platform for solid-state quantum information processing and nanoscale metrology. Of interest in these applications is the manipulation of the NV charge, which can be attained by optical excitation. Here, we use two-colour optical microscopy to investigate the dynamics of NV photo-ionization, charge diffusion and trapping in type-1b diamond. We combine fixed-point laser excitation and scanning fluorescence imaging to locally alter the concentration of negatively charged NVs, and to subsequently probe the corresponding redistribution of charge. We uncover the formation of spatial patterns of trapped charge, which we qualitatively reproduce via a model of the interplay between photo-excited carriers and atomic defects. Further, by using the NV as a probe, we map the relative fraction of positively charged nitrogen on localized optical excitation. These observations may prove important to transporting quantum information between NVs or to developing three-dimensional, charge-based memories.

Recovery of Utility Fixed Costs: Utility, Consumer, Environmental and Economist Perspectives

Energy Technology Data Exchange (ETDEWEB)

Wood, Lisa [Inst. for Electric Innovation and The Edison Foundation, Washington DC (United States); Hemphill, Ross [RCHemphill Solutions, Columbus, OH (United States); Howat, John [National Consumer Law Center, Boston, MA (United States); Cavanagh, Ralph [Natural Resources Defense Council, New York, NY (United States); Borenstein, Severin [Univ. of California, Berkeley, CA (United States); Deason, Jeff [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Schwartz, Lisa [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Schwartz, Lisa [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

2016-06-14

Utilities recover costs for providing electric service to retail customers through a combination of rate components that together comprise customers’ monthly electric bills. Rates and rate designs are set by state regulators and vary by jurisdiction, utility and customer class. In addition to the fundamental tenet of setting fair and reasonable rates, rate design balances economic efficiency, equity and fairness, customer satisfaction, utility revenue stability, and customer price and bill stability.1 At the most basic level, retail electricity bills in the United States typically include a fixed monthly customer charge — a set dollar amount regardless of energy usage — and a volumetric energy charge for each kilowatt-hour consumed.2 The energy charge may be flat across all hours, vary by usage level (for example, higher rates at higher levels of usage), or vary based on time of consumption.3 While some utility costs, such as fuel costs, clearly vary according to electricity usage, other costs are “fixed” over the short run — generally, those that do not vary over the course of a year. Depending on your point of view, and whether the state’s electricity industry has been restructured or remains vertically integrated, the set of costs that are “fixed” may be quite limited. Or the set may extend to all capacity costs for generation, transmission and distribution. In the long run, all costs are variable. In the context of flat or declining loads in some regions, utilities are proposing a variety of changes to retail rate designs, particularly for residential customers, to recover fixed costs. In this report, authors representing utility (Chapter 1), consumer (Chapter 2), environmentalist (Chapter 3) and economist (Chapter 4) perspectives discuss fixed costs for electric utilities and set out their principles for recovering those costs. The table on the next page summarizes each author’s relative preferences for various options for fixed cost

Synthesis of zeolites coal ash in surfactant modified in application and removal of orange 8 acid solution: study in batch, fixed bed column and evaluation ecotoxicological

International Nuclear Information System (INIS)

Magdalena, Carina Pitwak

2015-01-01

In this study, synthesized zeolitic material from coal ash and modified cationic surfactant was used for removing the acid dye Orange 8 (AL8) by adsorption process using moving bed and fixed-bed column. The raw material and adsorbents were characterized by different techniques, such as X-ray diffraction, X-ray fluorescence spectroscopy, among others. The adsorption of AL8 was performed by moving bed in order to optimize the results when they are launched in a fixed bed. The effects of adsorption on zeolite AL8 were compared: (1) Effect of counterions Br - and Cl - surfactant used in the modification of the zeolite; (2) effect of type of coal ash used as raw material in the synthesis of zeolites (fly and bottom). The following adsorbents were used in the study: fly and bottom zeolite modified by surfactant hexadecyltrimethylammonium bromide (ZLMS-Br-Br and ZPMS-Br) and fly zeolite modified by surfactant hexadecyltrimethylammonium chloride (ZLMS-Cl). The pseudo-second-order kinetic described the adsorption of the dye on all adsorbents. The equilibrium time was reached 40, 60 and 120 min for ZLMS-Br, ZLMS-Cl and ZPMS-Br, respectively. The adsorption equilibrium was analyzed by the equations of the models of linear and nonlinear isotherms of Langmuir, Freundlich, Temkin and Dubinin- Radushkevivh (DR) and the criterion of best fit was evaluated using the error functions.The DR model was adjusted better to the experimental data for the system AL8 / ZLMS-Br, the Freundlich model for AL8 / ZLMS-Cl and Langmuir for AL8 / ZPMS. According to the Langmuir maximum adsorption capacity was 4.67, 1.48 and 1.38 mg g -1 for ZLMS-Br, ZLMS-Cl and ZPMS-Br, in order. In studies employing fixed bed columns, the effects of inlet concentration (20- 30 mg L -1 ), flow rate (4.0 -5.3 mL min -1 ) and the bed height (5, 5 - 6.5 cm) above the breakthrough curves characteristics in the adsorption system were determined. The Adams-Bohart, Thomas, Yoon-Nelson models were applied to experimental

Modified denatured lysozyme effectively solubilizes fullerene c60 nanoparticles in water

Science.gov (United States)

Siepi, Marialuisa; Politi, Jane; Dardano, Principia; Amoresano, Angela; De Stefano, Luca; Monti, Daria Maria; Notomista, Eugenio

2017-08-01

Fullerenes, allotropic forms of carbon, have very interesting pharmacological effects and engineering applications. However, a very low solubility both in organic solvents and water hinders their use. Fullerene C60, the most studied among fullerenes, can be dissolved in water only in the form of nanoparticles of variable dimensions and limited stability. Here the effect on the production of C60 nanoparticles by a native and denatured hen egg white lysozyme, a highly basic protein, has been systematically studied. In order to obtain a denatured, yet soluble, lysozyme derivative, the four disulfides of the native protein were reduced and exposed cysteines were alkylated by 3-bromopropylamine, thus introducing eight additional positive charges. The C60 solubilizing properties of the modified denatured lysozyme proved to be superior to those of the native protein, allowing the preparation of biocompatible highly homogeneous and stable C60 nanoparticles using lower amounts of protein, as demonstrated by dynamic light scattering, transmission electron microscopy and atomic force microscopy studies. This lysozyme derivative could represent an effective tool for the solubilization of other carbon allotropes.

Hot-spot analysis to dissect the functional protein-protein interface of a tRNA-modifying enzyme.

Science.gov (United States)

Jakobi, Stephan; Nguyen, Tran Xuan Phong; Debaene, François; Metz, Alexander; Sanglier-Cianférani, Sarah; Reuter, Klaus; Klebe, Gerhard

2014-10-01

Interference with protein-protein interactions of interfaces larger than 1500 Ų by small drug-like molecules is notoriously difficult, particularly if targeting homodimers. The tRNA modifying enzyme Tgt is only functionally active as a homodimer. Thus, blocking Tgt dimerization is a promising strategy for drug therapy as this protein is key to the development of Shigellosis. Our goal was to identify hot-spot residues which, upon mutation, result in a predominantly monomeric state of Tgt. The detailed understanding of the spatial location and stability contribution of the individual interaction hot-spot residues and the plasticity of motifs involved in the interface formation is a crucial prerequisite for the rational identification of drug-like inhibitors addressing the respective dimerization interface. Using computational analyses, we identified hot-spot residues that contribute particularly to dimer stability: a cluster of hydrophobic and aromatic residues as well as several salt bridges. This in silico prediction led to the identification of a promising double mutant, which was validated experimentally. Native nano-ESI mass spectrometry showed that the dimerization of the suggested mutant is largely prevented resulting in a predominantly monomeric state. Crystal structure analysis and enzyme kinetics of the mutant variant further support the evidence for enhanced monomerization and provide first insights into the structural consequences of the dimer destabilization. © 2014 Wiley Periodicals, Inc.

Atom depth analysis delineates mechanisms of protein intermolecular interactions

International Nuclear Information System (INIS)

Alocci, Davide; Bernini, Andrea; Niccolai, Neri

2013-01-01

Highlights: •3D atom depth analysis is proposed to identify different layers in protein structures. •Amino acid contents for each layers have been analyzed for a large protein dataset. •Charged amino acids in the most external layer are present at very different extents. •Atom depth indexes of K residues reflect their side chains flexibility. •Mobile surface charges can be responsible for long range protein–protein recognition. -- Abstract: The systematic analysis of amino acid distribution, performed inside a large set of resolved protein structures, sheds light on possible mechanisms driving non random protein–protein approaches. Protein Data Bank entries have been selected using as filters a series of restrictions ensuring that the shape of protein surface is not modified by interactions with large or small ligands. 3D atom depth has been evaluated for all the atoms of the 2,410 selected structures. The amino acid relative population in each of the structural layers formed by grouping atoms on the basis of their calculated depths, has been evaluated. We have identified seven structural layers, the inner ones reproducing the core of proteins and the outer one incorporating their most protruding moieties. Quantitative analysis of amino acid contents of structural layers identified, as expected, different behaviors. Atoms of Q, R, K, N, D residues are increasingly more abundant in going from core to surfaces. An opposite trend is observed for V, I, L, A, C, and G. An intermediate behavior is exhibited by P, S, T, M, W, H, F and Y. The outer structural layer hosts predominantly E and K residues whose charged moieties, protruding from outer regions of the protein surface, reorient free from steric hindrances, determining specific electrodynamics maps. This feature may represent a protein signature for long distance effects, driving the formation of encounter complexes and the eventual short distance approaches that are required for protein–protein

Cellobiose Dehydrogenase Aryl Diazonium Modified Single Walled Carbon Nanotubes: Enhanced Direct Electron Transfer through a Positively Charged Surface

Science.gov (United States)

2011-01-01

One of the challenges in the field of biosensors and biofuel cells is to establish a highly efficient electron transfer rate between the active site of redox enzymes and electrodes to fully access the catalytic potential of the biocatalyst and achieve high current densities. We report on very efficient direct electron transfer (DET) between cellobiose dehydrogenase (CDH) from Phanerochaete sordida (PsCDH) and surface modified single walled carbon nanotubes (SWCNT). Sonicated SWCNTs were adsorbed on the top of glassy carbon electrodes and modified with aryl diazonium salts generated in situ from p-aminobenzoic acid and p-phenylenediamine, thus featuring at acidic pH (3.5 and 4.5) negative or positive surface charges. After adsorption of PsCDH, both electrode types showed excellent long-term stability and very efficient DET. The modified electrode presenting p-aminophenyl groups produced a DET current density of 500 μA cm−2 at 200 mV vs normal hydrogen reference electrode (NHE) in a 5 mM lactose solution buffered at pH 3.5. This is the highest reported DET value so far using a CDH modified electrode and comes close to electrodes using mediated electron transfer. Moreover, the onset of the electrocatalytic current for lactose oxidation started at 70 mV vs NHE, a potential which is 50 mV lower compared to when unmodified SWCNTs were used. This effect potentially reduces the interference by oxidizable matrix components in biosensors and increases the open circuit potential in biofuel cells. The stability of the electrode was greatly increased compared with unmodified but cross-linked SWCNTs electrodes and lost only 15% of the initial current after 50 h of constant potential scanning. PMID:21417322

Investigation of charges carrier density in phosphorus and boron doped SiNx:H layers for crystalline silicon solar cells

International Nuclear Information System (INIS)

Paviet-Salomon, B.; Gall, S.; Slaoui, A.

2013-01-01

Highlights: ► We investigate the properties of phosphorus and boron-doped silicon nitride films. ► Phosphorus-doped layers yield higher lifetimes than undoped ones. ► The fixed charges density decreases when increasing the films phosphorus content. ► Boron-doped films feature very low lifetimes. ► These doped layers are of particular interest for crystalline silicon solar cells. -- Abstract: Dielectric layers are of major importance in crystalline silicon solar cells processing, especially as anti-reflection coatings and for surface passivation purposes. In this paper we investigate the fixed charge densities (Q fix ) and the effective lifetimes (τ eff ) of phosphorus (P) and boron (B) doped silicon nitride layers deposited by plasma-enhanced chemical vapour deposition. P-doped layers exhibit a higher τ eff than standard undoped layers. In contrast, B-doped layers exhibit lower τ eff . A strong Q fix decrease is to be seen when increasing the P content within the film. Based on numerical simulations we also demonstrate that the passivation obtained with P- and B-doped layers are limited by the interface states rather than by the fixed charges

Swelling of whey and egg white protein hydrogels with stranded and particulate microstructures.

Science.gov (United States)

Li, Hui; Zhao, Lei; Chen, Xiao Dong; Mercadé-Prieto, Ruben

2016-02-01

Swelling of protein hydrogels in alkaline conditions strongly depends on the gel microstructure. Stranded transparent gels swell as predicted using a modified Flory-Rehner model with the net protein charge. Particulate opaque gels swell very differently, with a sudden increase at a narrow pH range. Its swelling is not controlled by the protein charge, but by the destruction of the non-covalent interactions. Comparable dissolution thresholds, one with pH and another with the degree of swelling, are observed in both types of microstructures. These conclusions are valid for both whey protein isolate (WPI) gels and egg white gels, suggesting that they are universal for all globular proteins that can form such microscructures. Differences are observed, however, from the prevalent chemical crosslinks in each protein system. Non-covalent interactions dominate WPI gels; when such interactions are destroyed at pH≥11.5 the gels swell extensively and eventually dissolve. In egg white gels, the higher degree of disulphide crosslinking allows extensive swelling when non-covalent interactions are destroyed, but dissolution only occurs at pH≥13 when covalent crosslinks are cleaved. The current study highlights that the microstructure of protein hydrogels, a unique particularity of protein systems compared to other synthetic hydrogels, defines swelling. Copyright © 2015 Elsevier B.V. All rights reserved.

A Power-Efficient Wireless Capacitor Charging System Through an Inductive Link.

Science.gov (United States)

Lee, Hyung-Min; Ghovanloo, Maysam

2013-10-01

A power-efficient wireless capacitor charging system for inductively powered applications has been presented. A bank of capacitors can be directly charged from an ac source by generating a current through a series charge injection capacitor and a capacitor charger circuit. The fixed charging current reduces energy loss in switches, while maximizing the charging efficiency. An adaptive capacitor tuner compensates for the resonant capacitance variations during charging to keep the amplitude of the ac input voltage at its peak. We have fabricated the capacitor charging system prototype in a 0.35- μ m 4-metal 2-poly standard CMOS process in 2.1 mm 2 of chip area. It can charge four pairs of capacitors sequentially. While receiving 2.7-V peak ac input through a 2-MHz inductive link, the capacitor charging system can charge each pair of 1 μ F capacitors up to ±2 V in 420 μ s, achieving a high measured charging efficiency of 82%.

Applying electric field to charged and polar particles between metallic plates: extension of the Ewald method.

Science.gov (United States)

Takae, Kyohei; Onuki, Akira

2013-09-28

We develop an efficient Ewald method of molecular dynamics simulation for calculating the electrostatic interactions among charged and polar particles between parallel metallic plates, where we may apply an electric field with an arbitrary size. We use the fact that the potential from the surface charges is equivalent to the sum of those from image charges and dipoles located outside the cell. We present simulation results on boundary effects of charged and polar fluids, formation of ionic crystals, and formation of dipole chains, where the applied field and the image interaction are crucial. For polar fluids, we find a large deviation of the classical Lorentz-field relation between the local field and the applied field due to pair correlations along the applied field. As general aspects, we clarify the difference between the potential-fixed and the charge-fixed boundary conditions and examine the relationship between the discrete particle description and the continuum electrostatics.

Inventory charge calculations in the nuclear fuel cycle

International Nuclear Information System (INIS)

Salmon, R.

1975-09-01

Simplified methods are presented for the calculation of inventory charges or carrying charges on fuel, which represent the indirect component of the fuel cycle cost. These methods permit rapid calculation of the changes in fuel cycle cost caused by changes in the amount or timing of fuel cycle expenditures. The methods are developed by applying the discounted cash flow procedure to a single batch of fuel. In typical cases, this would be a batch representing equilibrium or steady-state reactor operation. The cost equations used are the same as those used in the computer code REFCO, described in ORNL-4695, which was based on the discounted cash flow procedure with continuous discounting. Equivalent procedures using the fixed charge rate concept also are developed. This is done in such a way that consistency with the discounted cash flow procedure is maintained. The fixed charge rate used here is defined in terms of tax rates and the interest rates on debt and equity capital. An effective inventory time is also defined. This is a function of the lead or lag time, the interest rates on capital, and the exposure time of the batch. Tabulated values of the effective inventory time and other useful functions, such as the ratio of indirect to direct cost, are included. Cost calculations using these tables agree with those produced by REFCO, the accuracy being within 0.001 mill/kWhr in the cases studied. (U.S.)

Charge conserving current deposition scheme for PIC simulations in modified spherical coordinates

Science.gov (United States)

Cruz, F.; Grismayer, T.; Fonseca, R. A.; Silva, L. O.

2017-10-01

Global models of pulsar magnetospheres have been actively pursued in recent years. Both macro and microscopic (PIC) descriptions have been used, showing that collective processes of e-e + plasmas dominate the global structure of pulsar magnetospheres. Since these systems are best described in spherical coordinates, the algorithms used in cartesian simulations must be generalized. A problem of particular interest is that of charge conservation in PIC simulations. The complex geometry and irregular grids used to improve the efficiency of these algorithms represent major challenges in the design of a charge conserving scheme. Here we present a new first-order current deposition scheme for a 2D axisymmetric, log-spaced radial grid, that rigorously conserves charge. We benchmark this scheme in different scenarios, by integrating it with a spherical Yee scheme and Boris/Vay pushers. The results show that charge is conserved to machine precision, making it unnecessary to correct the electric field to guarantee charge conservation. This scheme will be particularly important for future studies aiming to bridge the microscopic physical processes of e-e + plasma generation due to QED cascades, its self-consistent acceleration and radiative losses to the global dynamics of pulsar magnetospheres. Work supported by the European Research Council (InPairs ERC-2015-AdG 695088), FCT (Portugal) Grant PD/BD/114307/2016, and the Calouste Gulbenkian Foundation through the 2016 Scientific Research Stimulus Program.

High-performance tandem organic light-emitting diodes based on a buffer-modified p/n-type planar organic heterojunction as charge generation layer

Science.gov (United States)

Wu, Yukun; Sun, Ying; Qin, Houyun; Hu, Shoucheng; Wu, Qingyang; Zhao, Yi

2017-04-01

High-performance tandem organic light-emitting diodes (TOLEDs) were realized using a buffer-modified p/n-type planar organic heterojunction (OHJ) as charge generation layer (CGL) consisting of common organic materials, and the configuration of this p/n-type CGL was "LiF/N,N'-diphenyl-N,N'-bis(1-napthyl)-1,1'-biphenyl-4,4'-diamine (NPB)/4,7-diphenyl-1,10-phenanthroline (Bphen)/molybdenum oxide (MoOx)". The optimized TOLED exhibited a maximum current efficiency of 77.6 cd/A without any out-coupling techniques, and the efficiency roll-off was greatly improved compared to the single-unit OLED. The working mechanism of the p/n-type CGL was discussed in detail. It is found that the NPB/Bphen heterojunction generated enough charges under a forward applied voltage and the carrier extraction was a tunneling process. These results could provide a new method to fabricate high-performance TOLEDs.

Application of Polarization Modulated Infrared Reflection Absorption Spectroscopy for electrocatalytic activity studies of laccase adsorbed on modified gold electrodes

International Nuclear Information System (INIS)

Olejnik, Piotr; Pawłowska, Aleksandra; Pałys, Barbara

2013-01-01

Orientation of the enzyme macromolecule on the electrode surface is crucially important for the efficiency of the electron transport between the active site and electrode surface. The orientation can be controlled by affecting the surface charge and the pH of the buffer solution. In this contribution we study laccase physically adsorbed on gold surface modified by mercapto-ethanol, lipid and variously charged diazonium salts. Polarization Modulated Infrared Reflection Absorption Spectroscopy (PMIRRAS) enables the molecular orientation study of the protein molecule by comparison of the amide I to amide II band intensity ratios assuming that the protein secondary structure does not change. We observe significant differences in the intensity ratios depending on the kind of support and the enzyme deposition. The comparison of infrared spectra and cyclic voltammetry responses of variously prepared laccase layers reveals that the parallel orientation of beta-sheet moieties results in high enzyme activity

Spectroscopic detection of fluorescent protein marker gene activity in genetically modified plants

Science.gov (United States)

Liew, O. W.; Chong, Jenny P. C.; Asundi, Anand K.

2005-04-01

This work focuses on developing a portable fibre optic fluorescence analyser for rapid identification of genetically modified plants tagged with a fluorescent marker gene. Independent transgenic tobacco plant lines expressing the enhanced green fluorescence protein (EGFP) gene were regenerated following Agrobacterium-mediated gene transfer. Molecular characterisation of these plant lines was carried out at the DNA level by PCR screening to confirm their transgenic status. Conventional transgene expression analysis was then carried out at the RNA level by RT-PCR and at the protein level by Western blotting using anti-GFP rabbit antiserum. The amount of plant-expressed EGFP on a Western blot was quantified against known amounts of purified EGFP by scanning densitometry. The expression level of EGFP in transformed plants was found to range from 0.1 - 0.6% of total extractable protein. A comparison between conventional western analysis of transformants and direct spectroscopic quantification using the fibre optic fluorescence analyser was made. The results showed that spectroscopic measurements of fluorescence emission from strong EGFP expressors correlated positively with Western blot data. However, the fluorescence analyser was also able to identify weakly expressing plant transformants below the detection limit of colorimetric Western blotting.

ABC's of pion charge exchange

International Nuclear Information System (INIS)

Gibbs, W.R.; Kaufmann, W.B.; Siegel, P.B.

1985-01-01

Calculations of pion single charge exchange using the PWIA and DWIA are presented. Emphasis is given to the effects of absorbtion and blocking. A microscopic calculation of the 0 0 excitation and low energy angular distribution is in excellent agreement with the data. A fixed nucleon multiple scattering calculation of the pion double charge exchange reaction is presented. Various valence neutron wave functions are used, and the contributions of different spatial orientations of the last two neutrons to the reaction are examined. The DCX cross section is found to be very sensitive to the inclusion of correlations in the two-neutron wave function. Satisfactory agreement with DCX data on 14 C can be obtained using a nucleonic picture of the nucleus

Charge-state correlated cross sections for the production of low-velocity highly charged Ne ions by heavy-ion bombardment

International Nuclear Information System (INIS)

Gray, T.J.; Cocke, C.L.; Justiniano, E.

1980-01-01

We report measured cross sections for the collisional production of highly charged low-velocity Ne recoil ions resulting from the bombardment of a thin Ne gas target by highly charged 1-MeV/amu C, N, O, and F projectiles. The measurements were made using time-of-flight techniques which allowed the simultaneous identification of the final charge state of both the low-velocity recoil ion and the high-velocity projectile for each collision event. For a given incident-projectile charge state, the recoil charge-state distribution is very dependent upon the final charge state of the projectile. Single- and double-electron capture events by incident bare nuclei and projectile K-shell ionization during the collision cause large shifts in the recoil charge-state distributions toward higher charge states. A previously proposed energy-deposition model is modified to include the effects of projectile charge-changing collisions during the collision for bare and hydrogenlike projectiles and is used to discuss the present experimental results

Physicochemical and immunologic characterization of low-molecular-weight allergoids of Dactylis glomerata pollen proteins.

Science.gov (United States)

Cirković, T D; Bukilica, M N; Gavrović, M D; Vujcić, Z M; Petrović, S; Jankov, R M

1999-02-01

Orchard grass (Dactylis glomerata) pollen proteins were chemically modified by means of acid anhydrides (maleic and succinic anhydride) to obtain low-molecular-weight allergoids. Chemical modification in both cases led to the replacement of one positive charge (epsilon amino group of Lys) by one negative charge, yielding proteins with changed physicochemical properties in comparison to the native orchard grass-pollen proteins. Physicochemical characterization of derivatives was done by gel chromatography, SDS-PAGE, and isoelectric focusing. To examine the IgE-binding properties of these derivatives, we carried out immunoblotting. To examine the ability of derivatives to induce IgG production, we immunized rabbits. Skin prick testing with the allergoids was performed on 15 individuals allergic to orchard grass pollens and on two healthy subjects. It was shown that the modified proteins retain their original molecular weights, but change pI to more acidic values. In the case of allergoids, a strong reduction in IgE binding was found. Immunization of rabbits with allergoids showed that the derivatives retain the ability to induce IgG production, and that the antisera obtained in such a way react to native (unmodified) extract. The ability of derivatives to induce allergic reaction was significantly reduced. The patients (86.6%) included in our study exhibited less than 50% of native extract response. Among them, 53.3% had no response to one or both allergoids. These modification procedures yield allergoids with a reduced allergenic activity and preserved immunogenic potential suitable for use in immunotherapy.

The fixABCX genes in Rhodospirillum rubrum encode a putative membrane complex participating in electron transfer to nitrogenase.

Science.gov (United States)

Edgren, Tomas; Nordlund, Stefan

2004-04-01

In our efforts to identify the components participating in electron transport to nitrogenase in Rhodospirillum rubrum, we used mini-Tn5 mutagenesis followed by metronidazole selection. One of the mutants isolated, SNT-1, exhibited a decreased growth rate and about 25% of the in vivo nitrogenase activity compared to the wild-type values. The in vitro nitrogenase activity was essentially wild type, indicating that the mutation affects electron transport to nitrogenase. Sequencing showed that the Tn5 insertion is located in a region with a high level of similarity to fixC, and extended sequencing revealed additional putative fix genes, in the order fixABCX. Complementation of SNT-1 with the whole fix gene cluster in trans restored wild-type nitrogenase activity and growth. Using Western blotting, we demonstrated that expression of fixA and fixB occurs only under conditions under which nitrogenase also is expressed. SNT-1 was further shown to produce larger amounts of both ribulose 1,5-bisphosphate carboxylase/oxygenase and polyhydroxy alkanoates than the wild type, indicating that the redox status is affected in this mutant. Using Western blotting, we found that FixA and FixB are soluble proteins, whereas FixC most likely is a transmembrane protein. We propose that the fixABCX genes encode a membrane protein complex that plays a central role in electron transfer to nitrogenase in R. rubrum. Furthermore, we suggest that FixC is the link between nitrogen fixation and the proton motive force generated in the photosynthetic reactions.

On the role of electrostatics on protein-protein interactions

Science.gov (United States)

Zhang, Zhe; Witham, Shawn; Alexov, Emil

2011-01-01

The role of electrostatics on protein-protein interactions and binding is reviewed in this article. A brief outline of the computational modeling, in the framework of continuum electrostatics, is presented and basic electrostatic effects occurring upon the formation of the complex are discussed. The role of the salt concentration and pH of the water phase on protein-protein binding free energy is demonstrated and indicates that the increase of the salt concentration tends to weaken the binding, an observation that is attributed to the optimization of the charge-charge interactions across the interface. It is pointed out that the pH-optimum (pH of optimal binding affinity) varies among the protein-protein complexes, and perhaps is a result of their adaptation to particular subcellular compartment. At the end, the similarities and differences between hetero- and homo-complexes are outlined and discussed with respect to the binding mode and charge complementarity. PMID:21572182

Pharmacological characterization of receptor-activity-modifying proteins (RAMPs) and the human calcitonin receptor.

Science.gov (United States)

Armour, S L; Foord, S; Kenakin, T; Chen, W J

1999-12-01

Receptor-activity-modifying proteins (RAMPs) are a family of single transmembrane domain proteins shown to be important for the transport and ligand specificity of the calcitonin gene-related peptide (CGRP) receptor. In this report, we describe the analysis of pharmacological properties of the human calcitonin receptor (hCTR) coexpressed with different RAMPs with the use of the Xenopus laevis melanophore expression system. We show that coexpression of RAMP3 with human calcitonin receptor changed the relative potency of hCTR to human calcitonin (hCAL) and rat amylin. RAMP1 and RAMP2, in contrast, had little effect on the change of hCTR potency to hCAL or rat amylin. When coexpressed with RAMP3, hCTR reversed the relative potency by a 3.5-fold loss in sensitivity to hCAL and a 19-fold increase in sensitivity to rat amylin. AC66, an inverse agonist, produced apparent simple competitive antagonism of hCAL and rat amylin, as indicated by linear Schild regressions. The potency of AC66 was changed in the blockade of rat amylin but not hCAL responses with RAMP3 coexpression. The mean pK(B) for AC66 to hCAL was 9.4 +/- 0.3 without RAMP3 and 9.45 +/- 0.07 with RAMP3. For the antagonism of AC66 to rat amylin, the pK(B) was 9.25 +/- 0.15 without RAMP3 and 8.2 +/- 0.35 with RAMP3. The finding suggests that RAMP3 might modify the active states of calcitonin receptor in such a way as to create a new receptor phenotype that is "amylin-like." Irrespective of the physiological association of the new receptor species, the finding that a coexpressed membrane protein can completely change agonist and antagonist affinities for a receptor raises implications for screening in recombinant receptor systems.

Lactose repressor protein modified with dansyl chloride: activity effects and fluorescence properties

International Nuclear Information System (INIS)

Hsieh, W.T.; Matthews, K.S.

1985-01-01

Chemical modification using 5-(dimethylamino)naphthalene-1-sulfonyl chloride (dansyl chloride) has been used to explore the importance of lysine residues involved in the binding activities of the lactose repressor and to introduce a fluorescent probe into the protein. Dansyl chloride modification of lac repressor resulted in loss of operator DNA binding at low molar ratios of reagent/monomer. Loss of nonspecific DNA binding was observed only at higher molar ratios, while isopropyl beta-D-thiogalactoside binding was not affected at any of the reagent levels studied. Lysine residues were the only modified amino acids detected. Protection of lysines-33 and -37 from modification by the presence of nonspecific DNA correlated with maintenance of operator DNA binding activity, and reaction of lysine-37 paralleled operator binding activity loss. Energy transfer between dansyl incorporated in the core region of the repressor protein and tryptophan-201 was observed, with an approximate distance of 23 A calculated between these two moieties

Compensating strong coupling with large charge

CERN Document Server

Alvarez-Gaume, Luis; Orlando, Domenico; Reffert, Susanne

2017-04-11

We study (conformal) field theories with global symmetries in the sector where the value of the global charge $Q$ is large. We find (as expected) that the low energy excitations of this sector are described by the general form of Goldstone's theorem in the non-relativistic regime. We also derive the unexpected result, first presented in [Hellerman:2015], that the effective field theory describing such sector of fixed $Q$ contains effective couplings $\\lambda_{\\text{eff}}\\sim \\lambda^b /Q^{a}$, where $\\lambda$ is the original coupling. Hence, large charge leads to weak coupling. In the last section of the paper we present an outline of how to compute anomalous dimensions in this limit.

Mucosal Immunogenicity of Genetically Modified Lactobacillus acidophilus Expressing an HIV-1 Epitope within the Surface Layer Protein.

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Akinobu Kajikawa

Full Text Available Surface layer proteins of probiotic lactobacilli are theoretically efficient epitope-displaying scaffolds for oral vaccine delivery due to their high expression levels and surface localization. In this study, we constructed genetically modified Lactobacillus acidophilus strains expressing the membrane proximal external region (MPER from human immunodeficiency virus type 1 (HIV-1 within the context of the major S-layer protein, SlpA. Intragastric immunization of mice with the recombinants induced MPER-specific and S-layer protein-specific antibodies in serum and mucosal secretions. Moreover, analysis of systemic SlpA-specific cytokines revealed that the responses appeared to be Th1 and Th17 dominant. These findings demonstrated the potential use of the Lactobacillus S-layer protein for development of oral vaccines targeting specific peptides.

The Protein-Sparing Modified Fast Diet

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Marwan Bakhach MD

2016-01-01

Full Text Available Objectives: The protein-sparing modified fast (PSMF is a rigorous way of rapidly losing a large amount of weight. Although adult studies have shown the PSMF to be effective, data in adolescents are lacking. The aim of this study was to determine the efficacy and safety of the PSMF in severely obese adolescents. Methods: 12 subjects who were evaluated in the Obesity Management Program at the Cleveland Clinic from 2011 to 2014 were included. The subjects were initiated on the PSMF after failing other conventional methods of weight loss. Once the goal weight was achieved, subjects were transitioned to the refeeding phase for weight maintenance. Results: Follow-up was scheduled at 3-month (11 patients and 6-month (6 patients intervals. At the 6-month follow-up visit, the average weight loss was 11.19 kg (95% confidence interval = -5.4, -27.8, P = .028, with average of 9.8% from baseline. Fifty percent of subjects had >5% weight loss and 20% had >10% weight loss. Four patients were lost to the follow-up (40%. An improvement was noted in total cholesterol and high-density lipoprotein. Due to a small sample size these results were not statistically significant. Side effects reported by subjects were mild dehydration due to nausea (2 patients, decreased energy (1 patient, and transient labile mood (1 patient. No life-threatening side effects were reported. Conclusion: Our results show that the PSMF diet can be used as an effective and safe method in the outpatient setting for rapid weight loss in adolescents with severe obesity.

Plasmons in inhomogeneously doped neutral and charged graphene nanodisks

Energy Technology Data Exchange (ETDEWEB)

Silveiro, Iván [ICFO-Institut de Ciencies Fotoniques, Mediterranean Technology Park, 08860 Castelldefels (Barcelona) (Spain); Javier García de Abajo, F., E-mail: javier.garciadeabajo@icfo.es [ICFO-Institut de Ciencies Fotoniques, Mediterranean Technology Park, 08860 Castelldefels (Barcelona) (Spain); ICREA-Institució Catalana de Recerca i Estudis Avançats, Passeig Lluís Companys, 23, 08010 Barcelona (Spain)

2014-03-31

We study plasmons in graphene nanodisks including the effect of inhomogeneity in the distribution of the doping charge. Specifically, we discuss the following two configurations: charged disks containing a fixed amount of additional carriers, which are self-consistently distributed along the surface to produce a uniform DC potential; and neutral disks exposed to a neighboring external point charge. A suitable finite-element method is elaborated to compute the charge density associated with the plasmons in the electrostatic limit. For charged disks, we find dipolar plasmons similar to those of uniformly doped graphene structures, in which the plasmon induced charge piles up near the edges. In contrast, in neutral disks placed near an external point charge, plasmons are strongly localized away from the edges. Surprisingly, a single external electron is enough to trap plasmons. The disks also display axially symmetric dark-plasmons, which can be excited through external illumination by coupling them to a neighboring metallic element. Our results have practical relevance for graphene nanophotonics under inhomogeneous doping conditions.

The negatively charged regions of lactoferrin binding protein B, an adaptation against anti-microbial peptides.

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Ari Morgenthau

Full Text Available Lactoferrin binding protein B (LbpB is a bi-lobed membrane bound lipoprotein that is part of the lactoferrin receptor complex in a variety of Gram-negative pathogens. Despite high sequence diversity among LbpBs from various strains and species, a cluster of negatively charged amino acids is invariably present in the protein's C-terminal lobe in all species except Moraxella bovis. The function of LbpB in iron acquisition has yet to be experimentally demonstrated, whereas in vitro studies have shown that LbpB confers protection against lactoferricin, a short cationic antimicrobial peptide released from the N- terminus of lactoferrin. In this study we demonstrate that the negatively charged regions can be removed from the Neisseria meningitidis LbpB without compromising stability, and this results in the inability of LbpB to protect against the bactericidal effects of lactoferricin. The release of LbpB from the cell surface by the autotransporter NalP reduces the protection against lactoferricin in the in vitro killing assay, attributed to removal of LbpB during washing steps, but is unlikely to have a similar impact in vivo. The protective effect of the negatively charged polysaccharide capsule in the killing assay was less than the protection conferred by LbpB, suggesting that LbpB plays a major role in protection against cationic antimicrobial peptides in vivo. The selective release of LbpB by NalP has been proposed to be a mechanism for evading the adaptive immune response, by reducing the antibody binding to the cell surface, but may also provide insights into the primary function of LbpB in vivo. Although TbpB and LbpB have been shown to be major targets of the human immune response, the selective release of LbpB suggests that unlike TbpB, LbpB may not be essential for iron acquisition, but important for protection against cationic antimicrobial peptides.

Dirac particles in the field of magnetic monopoles and of strong electric charges

International Nuclear Information System (INIS)

Schafer, A.; Muller, B.; Greiner, W.

1985-01-01

The field of a magnetic pointlike monopole acts in a similar way on a charged Dirac particle as the field of a very strong electric point charge. To explore this parallel it is constructed a field solution for an extended magnetic-charge distribution. In contrast to what is found for extended electric charges, the Hamiltonian remains nonself-adjoint for an extended magnetic monopole. This suggests that there exist a fundamental difference between the two cases. In particular, the appearance of undefined states for point monopoles is not a consequence of the mere strength of the magnetic-monopole charge, which has a minimum value fixed by Dirac's quantization condition

Positively-charged semi-tunnel is a structural and surface characteristic of polyphosphate-binding proteins: an in-silico study.

Directory of Open Access Journals (Sweden)

Zheng Zachory Wei

Full Text Available Phosphate is essential for all major life processes, especially energy metabolism and signal transduction. A linear phosphate polymer, polyphosphate (polyP, linked by high-energy phosphoanhydride bonds, can interact with various proteins, playing important roles as an energy source and regulatory factor. However, polyP-binding structures are largely unknown. Here we proposed a putative polyP binding site, a positively-charged semi-tunnel (PCST, identified by surface electrostatics analyses in polyP kinases (PPKs and many other polyP-related proteins. We found that the PCSTs in varied proteins were folded in different secondary structure compositions. Molecular docking calculations revealed a significant value for binding affinity to polyP in PCST-containing proteins. Utilizing the PCST identified in the β subunit of PPK3, we predicted the potential polyP-binding domain of PPK3. The discovery of this feature facilitates future searches for polyP-binding proteins and discovery of the mechanisms for polyP-binding activities. This should greatly enhance the understanding of the many physiological functions of protein-bound polyP and the involvement of polyP and polyP-binding proteins in various human diseases.

Parsimonious Charge Deconvolution for Native Mass Spectrometry

Science.gov (United States)

2018-01-01

Charge deconvolution infers the mass from mass over charge (m/z) measurements in electrospray ionization mass spectra. When applied over a wide input m/z or broad target mass range, charge-deconvolution algorithms can produce artifacts, such as false masses at one-half or one-third of the correct mass. Indeed, a maximum entropy term in the objective function of MaxEnt, the most commonly used charge deconvolution algorithm, favors a deconvolved spectrum with many peaks over one with fewer peaks. Here we describe a new “parsimonious” charge deconvolution algorithm that produces fewer artifacts. The algorithm is especially well-suited to high-resolution native mass spectrometry of intact glycoproteins and protein complexes. Deconvolution of native mass spectra poses special challenges due to salt and small molecule adducts, multimers, wide mass ranges, and fewer and lower charge states. We demonstrate the performance of the new deconvolution algorithm on a range of samples. On the heavily glycosylated plasma properdin glycoprotein, the new algorithm could deconvolve monomer and dimer simultaneously and, when focused on the m/z range of the monomer, gave accurate and interpretable masses for glycoforms that had previously been analyzed manually using m/z peaks rather than deconvolved masses. On therapeutic antibodies, the new algorithm facilitated the analysis of extensions, truncations, and Fab glycosylation. The algorithm facilitates the use of native mass spectrometry for the qualitative and quantitative analysis of protein and protein assemblies. PMID:29376659

The mechanism of lauric acid-modified protein nanocapsules escape from intercellular trafficking vesicles and its implication for drug delivery.

Science.gov (United States)

Jiang, Lijuan; Liang, Xin; Liu, Gan; Zhou, Yun; Ye, Xinyu; Chen, Xiuli; Miao, Qianwei; Gao, Li; Zhang, Xudong; Mei, Lin

2018-11-01

Protein nanocapsules have exhibited promising potential applications in the field of protein drug delivery. A major issue with various promising nano-sized biotherapeutics including protein nanocapsules is that owing to their particle size they are subject to cellular uptake via endocytosis, and become entrapped and then degraded within endolysosomes, which can significantly impair their therapeutic efficacy. In addition, many nano-sized biotherapeutics could be also sequestered by autophagosomes and degraded through the autolysosomal pathway. Thus, a limiting step in achieving an effective protein therapy is to facilitate the endosomal escape and auto-lysosomal escape to ensure cytosolic delivery of the protein drugs. Here, we prepared a protein nanocapsule based on BSA (nBSA) and the BSA nanocapsules modified with a bilayer of lauric acid (LA-nBSA) to investigate the escape effects from the endosome and autophagosome. The size distribution of nBSA and LA-nBSA analyzed using DLS presents a uniform diameter centered at 10 nm and 16 nm. The data also showed that FITC-labeled nBSA and LA-nBSA were taken up by the cells mainly through Arf-6-dependent endocytosis and Rab34-mediated macropinocytosis. In addition, LA-nBSA could efficiently escape from endosomal before the degradation in endo-lysosomes. Autophagy could also sequester the LA-nBSA through p62 autophagosome vesicles. These two types of nanocapsules underwent different intracellular destinies and lauric acid (LA) coating played a vital role in intracellular particle retention. In conclusion, the protein nanocapsules modified with LA could enhance the protein nanocapsules escape from intercellular trafficking vesicles, and protect the protein from degradation by the lysosomes.

Injectable nanocomposite cryogels for versatile protein drug delivery.

Science.gov (United States)

Koshy, Sandeep T; Zhang, David K Y; Grolman, Joshua M; Stafford, Alexander G; Mooney, David J

2018-01-01

Sustained, localized protein delivery can enhance the safety and activity of protein drugs in diverse disease settings. While hydrogel systems are widely studied as vehicles for protein delivery, they often suffer from rapid release of encapsulated cargo, leading to a narrow duration of therapy, and protein cargo can be denatured by incompatibility with the hydrogel crosslinking chemistry. In this work, we describe injectable nanocomposite hydrogels that are capable of sustained, bioactive, release of a variety of encapsulated proteins. Injectable and porous cryogels were formed by bio-orthogonal crosslinking of alginate using tetrazine-norbornene coupling. To provide sustained release from these hydrogels, protein cargo was pre-adsorbed to charged Laponite nanoparticles that were incorporated within the walls of the cryogels. The presence of Laponite particles substantially hindered the release of a number of proteins that otherwise showed burst release from these hydrogels. By modifying the Laponite content within the hydrogels, the kinetics of protein release could be precisely tuned. This versatile strategy to control protein release simplifies the design of hydrogel drug delivery systems. Here we present an injectable nanocomposite hydrogel for simple and versatile controlled release of therapeutic proteins. Protein release from hydrogels often requires first entrapping the protein in particles and embedding these particles within the hydrogel to allow controlled protein release. This pre-encapsulation process can be cumbersome, can damage the protein's activity, and must be optimized for each protein of interest. The strategy presented in this work simply premixes the protein with charged nanoparticles that bind strongly with the protein. These protein-laden particles are then placed within a hydrogel and slowly release the protein into the surrounding environment. Using this method, tunable release from an injectable hydrogel can be achieved for a variety of

Charge patterns as templates for the assembly of layered biomolecular structures.

Science.gov (United States)

Naujoks, Nicola; Stemmer, Andreas

2006-08-01

Electric fields are used to guide the assembly of biomolecules in predefined geometric patterns on solid substrates. Local surface charges serve as templates to selectively position proteins on thin-film polymeric electret layers, thereby creating a basis for site-directed layered assembly of biomolecular structures. Charge patterns are created using the lithographic capabilities of an atomic force microscope, namely by applying voltage pulses between a conductive tip and the sample. Samples consist of a poly(methyl methacrylate) layer on a p-doped silicon support. Subsequently, the sample is developed in a water-in-oil emulsion, consisting of a dispersed aqueous phase containing biotin-modified immunoglobulinG molecules, and a continuous nonpolar, insulating oil phase. The electrostatic fields cause a net force of (di)electrophoretic nature on the droplet, thereby guiding the proteins to the predefined locations. Due to the functionalization of the immunoglobulinG molecules with biotin-groups, these patterns can now be used to initiate the localized layer-by-layer assembly of biomolecules based on the avidin-biotin mechanism. By binding 40 nm sized biotin-labelled beads to the predefined locations via a streptavidin linker, we verify the functionality of the previously deposited immunoglobulinG-biotin. All assembly steps following the initial deposition of the immunoglobulinG from emulsion can conveniently be conducted in aqueous solutions. Results show that pattern definition is maintained after immersion into aqueous solution.

Experimental determination of net protein charge and A(tot) and K(a) of nonvolatile buffers in human plasma.

Science.gov (United States)

Staempfli, Henry R; Constable, Peter D

2003-08-01

The mechanism for an acid-base disturbance can be determined by using the strong ion approach, which requires species-specific values for the total concentration of plasma nonvolatile buffers (Atot) and the effective dissociation constant for plasma weak acids (Ka). The aim of this study was to experimentally determine Atot and Ka values for human plasma by using in vitro CO2 tonometry. Plasma Pco2 was systematically varied from 25 to 145 Torr at 37 degrees C, thereby altering plasma pH over the physiological range of 6.90-7.55, and plasma pH, Pco2, and concentrations of quantitatively important strong ions (Na+, K+, Ca2+, Mg2+, Cl-, lactate) and buffer ions (total protein, albumin, phosphate) were measured. Strong ion difference was estimated, and nonlinear regression was used to calculate Atot and Ka from the measured pH and Pco2 and estimated strong ion difference; the Atot and Ka values were then validated by using a published data set (Figge J, Rossing TH, and Fencl V, J Lab Clin Med 117: 453-467, 1991). The values (mean +/- SD) were as follows: Atot = 17.2 +/- 3.5 mmol/l (equivalent to 0.224 mmol/g of protein or 0.378 mmol/g of albumin); Ka = 0.80 +/- 0.60 x 10-7; negative log of Ka = 7.10. Mean estimates were obtained for strong ion difference (37 meq/l) and net protein charge (13+.0 meq/l). The experimentally determined values for Atot, Ka, and net protein charge should facilitate the diagnosis and treatment of acid-base disturbances in critically ill humans.

High Stability Pentacene Transistors Using Polymeric Dielectric Surface Modifier.

Science.gov (United States)

Wang, Xiaohong; Lin, Guangqing; Li, Peng; Lv, Guoqiang; Qiu, Longzhen; Ding, Yunsheng

2015-08-01

1,6-bis(trichlorosilyl)hexane (C6Cl), polystyrene (PS), and cross-linked polystyrene (CPS) were investigated as gate dielectric modified layers for high performance organic transistors. The influence of the surface energy, roughness and morphology on the charge transport of the organic thin-film transistors (OTFTs) was investigated. The surface energy and roughness both affect the grain size of the pentacene films which will control the charge carrier mobility of the devices. Pentacene thin-film transistors fabricated on the CPS modified dielectric layers exhibited charge carrier mobility as high as 1.11 cm2 V-1 s-1. The bias stress stability for the CPS devices shows that the drain current only decays 1% after 1530 s and the mobility never decreases until 13530 s.

A study of the influence of charged residues on β-hairpin formation by nuclear magnetic resonance and molecular dynamics.

Science.gov (United States)

Makowska, Joanna; Zmudzińska, Wioletta; Uber, Dorota; Chmurzyński, Lech

2014-12-01

Chain reversals are often nucleation sites in protein folding. The β-hairpins of FBP28 WW domain and IgG are stable and have been proved to initiate the folding and are, therefore, suitable for studying the influence of charged residues on β-hairpin conformation. In this paper, we carried out NMR examination of the conformations in solution of two fragments from the FPB28 protein (PDB code: 1E0L) (N-terminal part) namely KTADGKT-NH2 (1E0L 12-18, D7) and YKTADGKTY-NH2 (1E0L 11-19, D9), one from the B3 domain of the protein G (PDB code: 1IGD), namely DDATKT-NH2 (1IGD 51-56) (Dag1), and three variants of Dag1 peptide: DVATKT-NH2 (Dag2), OVATKT-NH2 (Dag3) and KVATKT-NH2 (Dag4), respectively, in which the original charged residue were replaced with non-polar residues or modified charged residues. It was found that both the D7 and D9 peptides form a large fraction bent conformations. However, no hydrophobic contacts between the terminal Tyr residues of D9 occur, which suggests that the presence of a pair of like-charged residues stabilizes chain reversal. Conversely, only the Dag1 and Dag2 peptides exhibit some chain reversal; replacing the second aspartic-acid residue with a valine and the first one with a basic residue results in a nearly extended conformation. These results suggest that basic residues farther away in sequence can result in stabilization of chain reversal owing to screening of the non-polar core. Conversely, smaller distance in sequence prohibits this screening, while the presence oppositely-charged residues can stabilize a turn because of salt-bridge formation.

Counterion release from a discretely charged surface in an electrolyte: Monte Carlo simulation study

International Nuclear Information System (INIS)

Hernández-Contreras, M

2015-01-01

Monte Carlo simulations allowed us to determine the amount of released electric charges from a discretely charged surface in 1:1 aqueous electrolyte solution as a function of surface charge density. Within the restricted primitive model and for a fixed concentration of 0.1 M bulk electrolyte in solution, there is an increase in the number of released counterions per unit surface area as the strength of the surface charge is enhanced. A similar behaviour of the number of released counterions was also found through the use of mean field and liquid theory methods

A thiophene-modified screen printed electrode for detection of dengue virus NS1 protein.

Science.gov (United States)

Silva, M M S; Dias, A C M S; Cordeiro, M T; Marques, E; Goulart, M O F; Dutra, R F

2014-10-01

A thiophene-modified screen printed electrode (SPE) for detection of the Dengue virus non-structural protein 1 (NS1), an important marker for acute phase diagnosis, is described. A sulfur-containing heterocyclic compound, the thiophene was incorporated to a carbon ink to prepare reproducible screen printed electrodes. After cured, the thiophene SPE was coated by gold nanoparticles conjugated to Protein A to form a nanostrutured surface. The Anti-NS1 antibodies immobilized via their Fc portions via Protein A, leaving their antigen specific sites free circumventing the problem of a random antibodies immobilization. Amperometric responses to the NS1 protein of dengue virus were obtained by cyclic voltammetries performed in presence of ferrocyanide/ferricyanide as redox probe. The calibration curve of immunosensor showed a linear response from 0.04 µg mL(-1) to 0.6 µg mL(-1) of NS1 with a good linear correlation (r=0.991, pink enhanced the electroanalytical properties of the SPEs, increasing their reproducibility and sensitivity. This point-of-care testing represents a great potential for use in epidemic situations, facilitating the early diagnosis in acute phase of dengue virus. Copyright © 2014 Elsevier B.V. All rights reserved.

Structure and stability of charged colloid-nanoparticle mixtures

Science.gov (United States)

Weight, Braden M.; Denton, Alan R.

2018-03-01

Physical properties of colloidal materials can be modified by addition of nanoparticles. Within a model of like-charged mixtures of particles governed by effective electrostatic interactions, we explore the influence of charged nanoparticles on the structure and thermodynamic phase stability of charge-stabilized colloidal suspensions. Focusing on salt-free mixtures of particles of high size and charge asymmetry, interacting via repulsive Yukawa effective pair potentials, we perform molecular dynamics simulations and compute radial distribution functions and static structure factors. Analysis of these structural properties indicates that increasing the charge and concentration of nanoparticles progressively weakens correlations between charged colloids. We show that addition of charged nanoparticles to a suspension of like-charged colloids can induce a colloidal crystal to melt and can facilitate aggregation of a fluid suspension due to attractive van der Waals interactions. We attribute the destabilizing influence of charged nanoparticles to enhanced screening of electrostatic interactions, which weakens repulsion between charged colloids. This interpretation is consistent with recent predictions of an effective interaction theory of charged colloid-nanoparticle mixtures.

Dielectric properties of proteins from simulations: tools and techniques

Science.gov (United States)

Simonson, Thomas; Perahia, David

1995-09-01

Tools and techniques to analyze the dielectric properties of proteins are described. Microscopic dielectric properties are determined by a susceptibility tensor of order 3 n, where n is the number of protein atoms. For perturbing charges not too close to the protein, the dielectric relaxation free energy is directly related to the dipole-dipole correlation matrix of the unperturbed protein, or equivalently to the covariance matrix of its atomic displacements. These are straightforward to obtain from existing molecular dynamics packages such as CHARMM or X- PLOR. Macroscopic dielectric properties can be derived from the dipolar fluctuations of the protein, by idealizing the protein as one or more spherical media. The dipolar fluctuations are again directly related to the covariance matrix of the atomic displacements. An interesting consequence is that the quasiharmonic approximation, which by definition exactly reproduces this covariance matrix, gives the protein dielectric constant exactly. Finally a technique is reviewed to obtain normal or quasinormal modes of vibration of symmetric protein assemblies. Using elementary group theory, and eliminating the high-frequency modes of vibration of each monomer, the limiting step in terms of memory and computation is finding the normal modes of a single monomer, with the other monomers held fixed. This technique was used to study the dielectric properties of the Tobacco Mosaic Virus protein disk.

Roles of Trm9- and ALKBH8-like proteins in the formation of modified wobble uridines in Arabidopsis tRNA

DEFF Research Database (Denmark)

Leihne, Vibeke; Kirpekar, Finn; Vågbø, Cathrine B

2011-01-01

Uridine at the wobble position of tRNA is usually modified, and modification is required for accurate and efficient protein translation. In eukaryotes, wobble uridines are modified into 5-methoxycarbonylmethyluridine (mcm(5)U), 5-carbamoylmethyluridine (ncm(5)U) or derivatives thereof. Here, we...... demonstrate, both by in vitro and in vivo studies, that the Arabidopsis thaliana methyltransferase AT1G31600, denoted by us AtTRM9, is responsible for the final step in mcm(5)U formation, thus representing a functional homologue of the Saccharomyces cerevisiae Trm9 protein. We also show that the enzymatic...... activity of AtTRM9 depends on either one of two closely related proteins, AtTRM112a and AtTRM112b. Moreover, we demonstrate that AT1G36310, denoted AtALKBH8, is required for hydroxylation of mcm(5)U to (S)-mchm(5)U in tRNA(Gly)(UCC), and has a function similar to the mammalian dioxygenase ALKBH8...

Label-free protein profiling of formalin-fixed paraffin-embedded (FFPE) heart tissue reveals immediate mitochondrial impairment after ionising radiation.

Science.gov (United States)

Azimzadeh, Omid; Scherthan, Harry; Yentrapalli, Ramesh; Barjaktarovic, Zarko; Ueffing, Marius; Conrad, Marcus; Neff, Frauke; Calzada-Wack, Julia; Aubele, Michaela; Buske, Christian; Atkinson, Michael J; Hauck, Stefanie M; Tapio, Soile

2012-04-18

Qualitative proteome profiling of formalin-fixed, paraffin-embedded (FFPE) tissue is advancing the field of clinical proteomics. However, quantitative proteome analysis of FFPE tissue is hampered by the lack of an efficient labelling method. The usage of conventional protein labelling on FFPE tissue has turned out to be inefficient. Classical labelling targets lysine residues that are blocked by the formalin treatment. The aim of this study was to establish a quantitative proteomics analysis of FFPE tissue by combining the label-free approach with optimised protein extraction and separation conditions. As a model system we used FFPE heart tissue of control and exposed C57BL/6 mice after total body irradiation using a gamma ray dose of 3 gray. We identified 32 deregulated proteins (p≤0.05) in irradiated hearts 24h after the exposure. The proteomics data were further evaluated and validated by bioinformatics and immunoblotting investigation. In good agreement with our previous results using fresh-frozen tissue, the analysis indicated radiation-induced alterations in three main biological pathways: respiratory chain, lipid metabolism and pyruvate metabolism. The label-free approach enables the quantitative measurement of radiation-induced alterations in FFPE tissue and facilitates retrospective biomarker identification using clinical archives. Copyright © 2012 Elsevier B.V. All rights reserved.

Mass and Angular Distributions of Charged Dihadron Production

Energy Technology Data Exchange (ETDEWEB)

Cummings, Mary Clare [Michigan U.

1990-01-01

Experiment 711, conducted at Fermilab. provided a unique handle towards understanding valence quark scattering by studying pairs of single. charged, high transverse momentum hadrons produced in collisions of 800 GeV /c protons on fixed metal targets. The apparatus consisted of a double-arm spectrometer. calorimetrically triggered. with high momentum resolution and a large angular acceptance for all charge states of particle pairs. The experiment was designed to select those hadron pairs that carrted most of the momentum and energy of the underlying scattered quarks and gluons. The charge of such "leading" hadrons is correlated with the charge of the quark that produced it. Quantum Chromodynamics (QCD) assumes that the scattering behavior of quarks ts independent of their charge, or "flavour": Experiment 711 could test this assumption. Tilis dissertation descrtbes the analysis of the mass and angular distributions of hadron pair production for three separate charge states: +-, ++ and --. The angular distributions are found to deviate from theory predictions of flavour symmetry. Also. the mass cross sections indicate ratios of positive to negative hard-scattered particles that are larger than expected from theory. These results could warrant reconsideration of the assumptions and approximations currently made in leading-order QCD calculations.

Isolation of a macrophage receptor for proteins modified by advanced glycosylation end products

International Nuclear Information System (INIS)

Radoff, S.; Vlassara, H.; Cerami, A.

1987-01-01

The nonenzymatic reaction of glucose with protein amino groups leads to the formation of irreversible AGE, such as the recently characterized glucose-derived crosslink, [2-furoyl-4(5)-(2-furanyl)-1-H-imidazole] (FFI). These products accumulate with time in aging tissues and diabetes, and are implicated in irreversible tissue damage. The authors have recently shown that macrophages bind and degrade AGE-proteins via a specific surface receptor, which is thus selectively removing senescent macromolecules. Scatchard plot analysis of binding data has indicated 1.5 x 10 5 receptors/cell with a binding affinity (Ka) of 1.7 x 10 7 /M. They have now isolated this receptor from murine macrophage RAW 264.7 membranes, solubilized with octylglucoside/protease inhibitors, and using FFI-Sepharose affinity chromatography and FPLC. The purified receptor binds radioactive FFI-containing compounds competitively. SDS-PAGE gels under reducing conditions indicate the receptor to be composed of two polypeptides, 83 Kda and 36 Kda. Crosslinking experiments with 125 I-AGE-albumin as ligand, indicate the 83 Kda subunit to be the AGE-binding peptide. These studies further characterize a macrophage receptor which selectively recognizes time-dependent glucose-modified proteins associated with aging and diabetes

Modified Adler sum rule and violation of charge symmetry

International Nuclear Information System (INIS)

Dominguez, C.A.; Moreno, H.; Zepeda, A.

The consequences of a once subtracted dispersion relation in the derivation of the Adler sum rule are investigated. It is shown that one can expect a breakdown of charge symmetry, of the isotriplet current hypothesis, and of scaling of the structure functions. These breakdowns are related to the possible presence of a non-zero subtraction function at asymptotic energies and arbitrary q 2 . Second class currents and PCAC relations are also discussed

Quark substructure approach to 4He charge distribution

International Nuclear Information System (INIS)

Wilets, L.; Alberg, M.A.; Pepin, S.; Stancu, F.; Carlson, J.; Koepf, W.

1997-01-01

We present a study of the 4 He charge distribution based on realistic nucleonic wave functions and incorporation of quark substructure. Any central depression of the proton point density seen in modern four-body calculations is too small by itself to lead to a correct description of the charge distribution of 4 He if folded with a fixed proton size parameter, as is usually done. We utilize six-quark structures calculated in the chromodielectric model for N-N interactions to find a swelling of the proton size as the internucleon distance decreases. This swelling is a result of the short-range dynamics in the N-N system. Using the independent pair approximation, the corresponding charge distribution of the proton is folded with the two-nucleon distribution generated from Green's function Monte Carlo calculations of the 4 He nucleonic wave function. We obtain a reasonably good fit to the experimental charge distribution of 4 He. Meson-exchange currents have not been included. copyright 1997 The American Physical Society

Charge State Coalescence During Electrospray Ionization Improves Peptide Identification by Tandem Mass Spectrometry

Science.gov (United States)

Meyer, Jesse G.; A. Komives, Elizabeth

2012-08-01

We report the effects of supercharging reagents dimethyl sulphoxide (DMSO) and m-nitrobenzyl alcohol ( m-NBA) applied to untargeted peptide identification, with special emphasis on non-tryptic peptides. Peptides generated from a mixture of five standard proteins digested with trypsin, elastase, or pepsin were separated with nanoflow liquid chromatography using mobile phases modified with either 5 % DMSO or 0.1 % m-NBA. Eluting peptides were ionized by online electrospray and sequenced by both CID and ETD using data-dependent MS/MS. Statistically significant improvements in peptide identifications were observed with DMSO co-solvent. In order to understand this observation, we assessed the effects of supercharging reagents on the chromatographic separation and the electrospray quality. The increase in identifications was not due to supercharging, which was greater for the 0.1 % m-NBA co-solvent and not observed for the 5.0 % DMSO co-solvent. The improved MS/MS efficiency using the DMSO modified mobile phase appeared to result from charge state coalescence.

Middle east respiratory syndrome coronavirus spike protein delivered by modified vaccinia virus ankara efficiently induces virus-neutralizing antibodies

NARCIS (Netherlands)

F. Song (Fei); R. Fux (Robert); L.B.V. Provacia (Lisette); A. Volz (Asisa); M. Eickmann; S. Becker (Stephan); A.D.M.E. Osterhaus (Albert); B.L. Haagmans (Bart); G. Suttera (Gerd)

2013-01-01

textabstractMiddle East respiratory syndrome coronavirus (MERS-CoV) has recently emerged as a causative agent of severe respiratory disease in humans. Here, we constructed recombinant modified vaccinia virus Ankara (MVA) expressing full-length MERS-CoV spike (S) protein (MVA-MERS-S). The genetic

Turning an energy analyzer into a mass spectrometer and a charge sorter

International Nuclear Information System (INIS)

Lee, K.

2004-01-01

Full text:When a swarm of ions are accelerated by a pulsed electric field for a common duration before entering an electrostatically dispersive energy analyzer, they will be sorted according to their charge-to-mass ratio q/m. In other words, the apparent kinetic energy upon which an ion will be registered in an apparent 'energy' spectrum thus obtained is proportional to its q/m ratio. For ions of a fixed mass m, the apparent energy spectrum becomes a charge state distribution spectrum. For ions of a fixed charge q, the apparent energy spectrum becomes a mass spectrum. In essence, an energy analyzer becomes both a charge sorter and a mass spectrometer when operated in this mode. When this technique is brought to bear on the detection of photofragment ions, a distinct advantage emerges. Kinetic energy release distribution (KERD) of fragment ions conventionally is extracted from fitting of the ion time-of-flight profile to ion trajectory simulation results, as such a fair amount of uncertainty exists in the KERD thus obtained. With the current novel approach to mass spectrometry, the KERDs of photofragment ions can be obtained directly, and as a result, with a greater accuracy. This should advance the study of fragmentation dynamics considerably

Studies on utilization of nitrogen-fixing bacteria for saving energy; Chisso koteikin no katsuyo ni yoru sho energy no kenkyu

Energy Technology Data Exchange (ETDEWEB)

Uozumi, T; Koyama, R; Horiuchi, M; Hidaka, M; Masaki, H [The University of Tokyo, Tokyo (Japan); Shigematsu, T; Inoue, A [New Energy and Industrial Technology Development Organization, Tokyo, (Japan)

1997-02-01

This paper describes analysis and enhancement of nitrogen-fixing gene of rice root bacteria, such as Klebsiella oxytoca, Azospirillum lipoferumn and Sphingomonas paucimobilis, for realizing energy saving through conservation of nitrogenous fertilizers. For K. oxytoca, modified strain R-16 was developed, which can fix nitrogen effectively even in the presence of NH4{sup +}. Nitrogen-fixing ability of A. lipoferumn depends on the activity control by the modification of nitrogen-fixing enzyme as well as on the adjustment of transcription level by the transcription activating gene, nifA. The control gene relating to the above was analyzed by making clones. As a result, a modified strain TAl without the control by NH4{sup +} was developed. The R-16 and TAl strains were inoculated into rice sterile-cultured without nitrogen. Consequently, inoculated strains were settled in the root, which resulted in the increased vegetation weight of plant to two times heavier than that without inoculation. 9 refs.

Colon luminal content and epithelial cell morphology are markedly modified in rats fed with a high-protein diet

OpenAIRE

Andriamihaja, Mireille; Davila-Gay, Anne-Marie; Eklou, Mamy; Petit, Nathalie; Delpal, Serge; Allek, Fadhila; Blais, Anne; Delteil, Corine; Tomé, Daniel; Blachier, Francois

2010-01-01

Andriamihaja M, Davila A, Eklou-Lawson M, Petit N, Delpal S, Allek F, Blais A, Delteil C, Tome D, Blachier F. Colon luminal content and epithelial cell morphology are markedly modified in rats fed with a high-protein diet. Am J Physiol Gastrointest Liver Physiol 299: G1030-G1037, 2010. First published August 5, 2010; doi: 10.1152/ajpgi.00149.2010.-Hyperproteic diets are used in human nutrition to obtain body weight reduction. Although increased protein ingestion results in an increased transf...

N(epsilon)-carboxymethyllysine-modified proteins are unable to bind to RAGE and activate an inflammatory response.

Science.gov (United States)

Buetler, Timo M; Leclerc, Estelle; Baumeyer, Alexandra; Latado, Helia; Newell, John; Adolfsson, Oskar; Parisod, Véronique; Richoz, Janique; Maurer, Sarah; Foata, Francis; Piguet, Dominique; Junod, Sylviane; Heizmann, Claus W; Delatour, Thierry

2008-03-01

Advanced glycation endproducts (AGEs) containing carboxymethyllysine (CML) modifications are generally thought to be ligands of the receptor for AGEs, RAGEs. It has been argued that this results in the activation of pro-inflammatory pathways and diseases. However, it has not been shown conclusively that a CML-modified protein can interact directly with RAGE. Here, we have analyzed whether beta-lactoglobulin (bLG) or human serum albumin (HSA) modified chemically to contain only CML (10-40% lysine modification) can (i) interact with RAGE in vitro and (ii) interact with and activate RAGE in lung epithelial cells. Our results show that CML-modified bLG or HSA are unable to bind to RAGE in a cell-free assay system (Biacore). Furthermore, they are unable to activate pro-inflammatory signaling in the cellular system. Thus, CML probably does not form the necessary structure(s) to interact with RAGE and activate an inflammatory signaling cascade in RAGE-expressing cells.

Pepsin immobilized in dextran-modified fused-silica capillaries for on-line protein digestion and peptide mapping

Energy Technology Data Exchange (ETDEWEB)

Stigter, E.C.A. [Division of Biomedical Analysis, Department of Pharmaceutical Sciences, Faculty of Science, Utrecht University, Sorbonnelaan 16, 3584 CA Utrecht (Netherlands)], E-mail: e.c.a.stigter@uu.nl; Jong, G.J. de; Bennekom, W.P. van [Division of Biomedical Analysis, Department of Pharmaceutical Sciences, Faculty of Science, Utrecht University, Sorbonnelaan 16, 3584 CA Utrecht (Netherlands)

2008-07-07

On-line digestion of proteins under acidic conditions was studied using micro-reactors consisting of dextran-modified fused-silica capillaries with covalently immobilized pepsin. The proteins used in this study differed in molecular weight, isoelectric point and sample composition. The injected protein samples were completely digested in 3 min and the digest was analyzed with micro-high performance liquid chromatography (HPLC) and tandem mass spectrometry (MS/MS). The different proteins present in the samples could be identified with a Mascot database search on the basis of auto-MS/MS data. It proved also to be possible to digest and analyze protein mixtures with a sequence coverage of 55% and 97% for the haemoglobin {beta}- and {alpha}-chain, respectively, and 35-55% for the various casein variants. Protease auto-digestion, sample carry-over and loss of signal due to adsorption of the injected proteins were not observed. The backpressure of the reactor is low which makes coupling to systems such as Surface Plasmon Resonance biosensors, which do not tolerate too high pressure, possible. The reactor was stable for at least 40 days when used continuously.

Pepsin immobilized in dextran-modified fused-silica capillaries for on-line protein digestion and peptide mapping.

Science.gov (United States)

Stigter, E C A; de Jong, G J; van Bennekom, W P

2008-07-07

On-line digestion of proteins under acidic conditions was studied using micro-reactors consisting of dextran-modified fused-silica capillaries with covalently immobilized pepsin. The proteins used in this study differed in molecular weight, isoelectric point and sample composition. The injected protein samples were completely digested in 3 min and the digest was analyzed with micro-high performance liquid chromatography (HPLC) and tandem mass spectrometry (MS/MS). The different proteins present in the samples could be identified with a Mascot database search on the basis of auto-MS/MS data. It proved also to be possible to digest and analyze protein mixtures with a sequence coverage of 55% and 97% for the haemoglobin beta- and alpha-chain, respectively, and 35-55% for the various casein variants. Protease auto-digestion, sample carry-over and loss of signal due to adsorption of the injected proteins were not observed. The backpressure of the reactor is low which makes coupling to systems such as Surface Plasmon Resonance biosensors, which do not tolerate too high pressure, possible. The reactor was stable for at least 40 days when used continuously.

Ionic liquid-modified metal sulfides/graphene oxide nanocomposites for photoelectric conversion

International Nuclear Information System (INIS)

Zhang, Yu; Zhang, Yù; Pei, Qi; Feng, Ting; Mao, Hui; Zhang, Wei; Wu, Shuyao; Liu, Daliang; Wang, Hongyu; Song, Xi-Ming

2015-01-01

Graphical abstract: - Highlights: • Metal sulfide (CdS, ZnS, Ag 2 S)/GO nanocomposites were prepared by electrostatic adherence. • Ionic liquid was used to link the metal sulfide and GO in the electrostatic adherence process. • The as-prepared samples showed enhanced photocurrent and highly efficient photocatalytic activity under visible light irradiation. - Abstract: Ionic liquid-modified metal sulfides/graphene oxide nanocomposites are prepared via a facile electrostatic adsorption. Ionic liquid (IL) is firstly used as surface modifier and structure-directing agent of metal sulfide (MS) crystallization process, obtaining ionic liquid modified-MS (IL-MS) nanoparticles with positive charges on surface. IL-MS/GO is obtained by electrostatic adherence between positively charged IL-MS and negatively charged graphene oxide (GO). The as-prepared sample shows enhanced photocurrent and highly efficient photocatalytic activity under visible light irradiation, indicating IL-MS/GO nanocomposites greatly promoted the separation of photogenerated electron–hole pairs

Research on cylinder processes of gasoline homogenous charge compression ignition (HCCI) engine

Science.gov (United States)

Cofaru, Corneliu

2017-10-01

This paper is designed to develop a HCCI engine starting from a spark ignition engine platform. The engine test was a single cylinder, four strokes provided with carburetor. The results of experimental research on this version were used as a baseline for the next phase of the work. After that, the engine was modified for a HCCI configuration, the carburetor was replaced by a direct fuel injection system in order to control precisely the fuel mass per cycle taking into account the measured intake air-mass. To ensure that the air - fuel mixture auto ignite, the compression ratio was increased from 9.7 to 11.5. The combustion process in HCCI regime is governed by chemical kinetics of mixture of air-fuel, rein ducted or trapped exhaust gases and fresh charge. To modify the quantities of trapped burnt gases, the exchange gas system was changed from fixed timing to variable valve timing. To analyze the processes taking place in the HCCI engine and synthesizing a control system, a model of the system which takes into account the engine configuration and operational parameters are needed. The cylinder processes were simulated on virtual model. The experimental research works were focused on determining the parameters which control the combustion timing of HCCI engine to obtain the best energetic and ecologic parameters.

Implications of protein polymorphism on protein phase behaviour

NARCIS (Netherlands)

Stegen, J.; Schoot, van der P.P.A.M.

2015-01-01

The phase behaviour of small globular proteins is often modeled by approximating them as spherical particles with fixed internal structure. However, changes in the local environment of a protein can lead to changes in its conformation rendering this approximation invalid. We present a simple

Dual Mode Fluorophore-Doped Nickel Nitrilotriacetic Acid-Modified Silica Nanoparticles Combine Histidine-Tagged Protein Purification with Site-Specific Fluorophore Labeling

OpenAIRE

Kim, Sung Hoon; Jeyakumar, M.; Katzenellenbogen, John A.

2007-01-01

We present the first example of a fluorophore-doped nickel chelate surface- modified silica nanoparticle that functions in a dual mode, combining histidine-tagged protein purification with site-specific fluorophore labeling. Tetramethylrhodamine (TMR)-doped silica nanoparticles, estimated to contain 700–900 TMRs per ca. 23-nm particle, were surface modified with nitrilotriacetic acid (NTA), producing TMR-SiO2-NTA-Ni+2. Silica-embedded TMR retains very high quantum yield, is resistant to quenc...

Characterization of the anti-HIV effects of native lactoferrin and other milk proteins and protein-derived peptides

NARCIS (Netherlands)

Berkhout, Ben; van Wamel, Jeroen L. B.; Beljaars, Leonie; Meijer, Dirk K. F.; Visser, Servaas; Floris, René

2002-01-01

In a search for natural proteins with anti-HIV activity, we screened a large set of purified proteins from bovine milk and peptide fragments thereof. Because several charged proteins and peptides are known to inhibit the process of virus entry, we selected proteins with an unusual charge composition

Characterization of the anti-HIV effects of native lactoferrin and other milk proteins and protein-derived peptides

NARCIS (Netherlands)

Berkhout, B; van Wamel, JLB; Beljaars, L; Meijer, DKF; Visser, Servaas; Floris, R

In a search for natural proteins with anti-HIV activity, we screened a large set of purified proteins from bovine milk and peptide fragments thereof. Because several charged proteins and peptides are known to inhibit the process of virus entry, we selected proteins with an unusual charge composition

Electrochemical immunosensors for the detection of survival motor neuron (SMN) protein using different carbon nanomaterials-modified electrodes.

Science.gov (United States)

Eissa, Shimaa; Alshehri, Nawal; Rahman, Anas M Abdel; Dasouki, Majed; Abu-Salah, Khalid M; Zourob, Mohammed

2018-03-15

Spinal muscular atrophy is an untreatable potentially fatal hereditary disorder caused by loss-of-function mutations in the survival motor neuron (SMN) 1 gene which encodes the SMN protein. Currently, definitive diagnosis relies on the demonstration of biallelic pathogenic variants in SMN1 gene. Therefore, there is an urgent unmet need to accurately quantify SMN protein levels for screening and therapeutic monitoring of symptomatic newborn and SMA patients, respectively. Here, we developed a voltammetric immunosensor for the sensitive detection of SMN protein based on covalently functionalized carbon nanofiber-modified screen printed electrodes. A comparative study of six different carbon nanomaterial-modified electrodes (carbon, graphene (G), graphene oxide (GO), single wall carbon nanotube (SWCNT), multi-wall carbon nanotube (MWCNT), and carbon nanofiber (CNF)) was performed. 4-carboxyphenyl layers were covalently grafted on the six electrodes by electroreduction of diazonium salt. Then, the terminal carboxylic moieties on the electrodes surfaces were utilized to immobilize the SMN antibody via EDC/NHS chemistry and to fabricate the immunosensors. The electrochemical characterization and analytical performance of the six immunosensors suggest that carbon nanofiber is a better electrode material for the SMN immunosensor. The voltammetric SMN carbon nanofiber-based immunosensor showed high sensitivity (detection limit of 0.75pg/ml) and selectivity against other proteins such as cystic fibrosis transmembrane conductance regulator (CFTR) and dystrophin (DMD). We suggest that this novel biosensor is superior to other developed assays for SMN detection in terms of lower cost, higher sensitivity, simplicity and capability of high throughput screening. Copyright © 2017 Elsevier B.V. All rights reserved.

Utilizing whey protein isolate and polysaccharide complexes to stabilize aerated dairy gels.

Science.gov (United States)

O'Chiu, Emily; Vardhanabhuti, Bongkosh

2017-05-01

Heated soluble complexes of whey protein isolate (WPI) with polysaccharides may be used to modify the properties of aerated dairy gels, which could be formulated into novel-textured high-protein desserts. The objective of this study was to determine the effect of polysaccharide charge density and concentration within a WPI-polysaccharide complex on the physical properties of aerated gels. Three polysaccharides having different degrees of charge density were chosen: low-methoxyl pectin, high-methoxyl type D pectin, and guar gum. Heated complexes were prepared by heating the mixed dispersions (8% protein, 0 to 1% polysaccharide) at pH 7. To form aerated gels, 2% glucono-δ-lactone was added to the dispersions of skim milk powder and heated complex and foam was generated by whipping with a handheld frother. The foam set into a gel as the glucono-δ-lactone acidified to a final pH of 4.5. The aerated gels were evaluated for overrun, drainage, gel strength, and viscoelastic properties. Without heated complexes, stable aerated gels could not be formed. Overrun of aerated gel decreased (up to 73%) as polysaccharide concentration increased from 0.105 to 0.315% due to increased viscosity, which limited air incorporation. A negative relationship was found between percent drainage and dispersion viscosity. However, plotting of drainage against dispersion viscosity separated by polysaccharide type revealed that drainage decreased most in samples with high-charge-density, low-methoxyl pectin followed by those with low-charge-density, high-methoxyl type D pectin. Aerated gels with guar gum (no charge) did not show improvement to stability. Rheological results showed no significant difference in gelation time among samples; therefore, stronger interactions between WPI and high-charge-density polysaccharide were likely responsible for increased stability. Stable dairy aerated gels can be created from WPI-polysaccharide complexes. High-charge-density polysaccharides, at

Compensating strong coupling with large charge

Energy Technology Data Exchange (ETDEWEB)

Alvarez-Gaume, Luis [Theory Department - CERN,CH-1211 Geneva 23 (Switzerland); Simons Center for Geometry and Physics, State University of New York,Stony Brook, NY-11794-3636 (United States); Loukas, Orestis; Orlando, Domenico; Reffert, Susanne [Albert Einstein Center for Fundamental Physics,Institute for Theoretical Physics, University of Bern,Sidlerstrasse 5, CH-3012 Bern (Switzerland)

2017-04-11

We study some (conformal) field theories with global symmetries in the sector where the value of the global charge Q is large. We find (as expected) that the low energy excitations of this sector are described by the general form of Goldstone’s theorem in the non-relativistic regime. We also derive the unexpected result, first presented in https://www.doi.org/10.1007/JHEP12(2015)071, that the effective field theory describing such sector of fixed Q contains effective couplings λ{sub eff}∼λ{sup b}/Q{sup a}, where λ is the original coupling. Hence, large charge leads to weak coupling. In the last section of the paper we present an outline of how to compute anomalous dimensions of the O(n) model in this limit.

Identification and modulation of the key amino acid residue responsible for the pH sensitivity of neoculin, a taste-modifying protein.

Directory of Open Access Journals (Sweden)

Ken-ichiro Nakajima

Full Text Available Neoculin occurring in the tropical fruit of Curculigo latifolia is currently the only protein that possesses both a sweet taste and a taste-modifying activity of converting sourness into sweetness. Structurally, this protein is a heterodimer consisting of a neoculin acidic subunit (NAS and a neoculin basic subunit (NBS. Recently, we found that a neoculin variant in which all five histidine residues are replaced with alanine elicits intense sweetness at both neutral and acidic pH but has no taste-modifying activity. To identify the critical histidine residue(s responsible for this activity, we produced a series of His-to-Ala neoculin variants and evaluated their sweetness levels using cell-based calcium imaging and a human sensory test. Our results suggest that NBS His11 functions as a primary pH sensor for neoculin to elicit taste modification. Neoculin variants with substitutions other than His-to-Ala were further analyzed to clarify the role of the NBS position 11 in the taste-modifying activity. We found that the aromatic character of the amino acid side chain is necessary to elicit the pH-dependent sweetness. Interestingly, since the His-to-Tyr variant is a novel taste-modifying protein with alternative pH sensitivity, the position 11 in NBS can be critical to modulate the pH-dependent activity of neoculin. These findings are important for understanding the pH-sensitive functional changes in proteinaceous ligands in general and the interaction of taste receptor-taste substance in particular.

Dual-mode fluorophore-doped nickel nitrilotriacetic acid-modified silica nanoparticles combine histidine-tagged protein purification with site-specific fluorophore labeling.

Science.gov (United States)

Kim, Sung Hoon; Jeyakumar, M; Katzenellenbogen, John A

2007-10-31

We present the first example of a fluorophore-doped nickel chelate surface-modified silica nanoparticle that functions in a dual mode, combining histidine-tagged protein purification with site-specific fluorophore labeling. Tetramethylrhodamine (TMR)-doped silica nanoparticles, estimated to contain 700-900 TMRs per ca. 23 nm particle, were surface modified with nitrilotriacetic acid (NTA), producing TMR-SiO2-NTA-Ni2+. Silica-embedded TMR retains very high quantum yield, is resistant to quenching by buffer components, and is modestly quenched and only to a certain depth (ca. 2 nm) by surface-attached Ni2+. When exposed to a bacterial lysate containing estrogen receptor alpha ligand binding domain (ERalpha) as a minor component, these beads showed very high specificity binding, enabling protein purification in one step. The capacity and specificity of these beads for binding a his-tagged protein were characterized by electrophoresis, radiometric counting, and MALDI-TOF MS. ERalpha, bound to TMR-SiO2-NTA-Ni++ beads in a site-specific manner, exhibited good activity for ligand binding and for ligand-induced binding to coactivators in solution FRET experiments and protein microarray fluorometric and FRET assays. This dual-mode type TMR-SiO2-NTA-Ni2+ system represents a powerful combination of one-step histidine-tagged protein purification and site-specific labeling with multiple fluorophore species.

Preparation of C-terminally modified chemokines by expressed protein ligation.

Science.gov (United States)

Baumann, Lars; Steinhagen, Max; Beck-Sickinger, Annette G

2013-01-01

In order to link structural features on a molecular level to the function of chemokines, site-specific modification strategies are strongly required. These can be used to incorporate fluorescent dyes and/or physical probes to allow investigations in a wide range of biological and physical techniques, e.g., nuclear magnetic resonance (NMR) spectroscopy, fluorescence microscopy, fluorescence resonance energy transfer (FRET), or fluorescence correlation spectroscopy (FCS). Only a limited number of functional groups within the 20 canonical amino acids allow ligation strategies that can be helpful to introduce novel functionalities, which in turn expand the scope of chemoselective and orthogonal reactivity of (semi)synthetic chemokines. In the present chapter we mainly focus on the fabulous history of native chemical ligation (NCL) and provide a general protocol for the preparation of C-terminally modified SDF-1α including tips and tricks for practical work. We believe that this protocol can be easily adapted to other chemokines and many proteins in general.

Local Two-Photon Couplings and the J=0 Fixed Pole in Real and Virtual Compton Scattering

Energy Technology Data Exchange (ETDEWEB)

Brodsky, Stanley J.; Llanes-Estrada, Felipe J.; Szczepaniak, Adam P.

2008-12-05

The local coupling of two photons to the fundamental quark currents of a hadron gives an energy-independent contribution to the Compton amplitude proportional to the charge squared of the struck quark, a contribution which has no analog in hadron scattering reactions. We show that this local contribution has a real phase and is universal, giving the same contribution for real or virtual Compton scattering for any photon virtuality and skewness at fixed momentum transfer squared t. The t-dependence of this J = 0 fixed Regge pole is parameterized by a yet unmeasured even charge-conjugation form factor of the target nucleon. The t = 0 limit gives an important constraint on the dependence of the nucleon mass on the quark mass through the Weisberger relation. We discuss how this 1=x form factor can be extracted from high energy deeply virtual Compton scattering and examine predictions given by models of the H generalized parton distribution.

Investigation of the electrochemical and electrocatalytic behavior of positively charged gold nanoparticle and L-cysteine film on an Au electrode

International Nuclear Information System (INIS)

Zhang Lingyan; Yuan Ruo; Chai Yaqing; Li Xuelian

2007-01-01

Positively charged gold nanoparticle (positively charged nano-Au), which was prepared, characterized by ξ-potential and transmission electron microscopy (TEM) was used in combination with L-cysteine to fabricate a modified electrode for electrocatalytic reaction of biomolecules. Compared with electrodes modified by negatively charged gold nanoparticle/L-cysteine, or L-cysteine alone, the electrode modified by the positively charged gold nanoparticle/L-cysteine exhibited excellent electrochemical behavior toward the oxidation of biomolecules such as ascorbic acid, dopamine and hydrogen peroxide. Moreover, the proposed mechanism for electrocatalytic response of positively charged gold nanoparticle was discussed. The immunosensor showed a specific to ascorbic acid in the range 5.1 x 10 -7 -6.7 x 10 -4 M and a low detection limit of 1.5 x 10 -7 M. The experimental results demonstrate that positively charged gold nanoparticle have more efficient electrocatalytic reaction than negatively charged gold nanoparticle, which opens up new approach for fabricating sensor

The Role of Histone Protein Modifications and Mutations in Histone Modifiers in Pediatric B-Cell Progenitor Acute Lymphoblastic Leukemia

Science.gov (United States)

Janczar, Szymon; Janczar, Karolina; Pastorczak, Agata; Harb, Hani; Paige, Adam J. W.; Zalewska-Szewczyk, Beata; Danilewicz, Marian; Mlynarski, Wojciech

2017-01-01

While cancer has been long recognized as a disease of the genome, the importance of epigenetic mechanisms in neoplasia was acknowledged more recently. The most active epigenetic marks are DNA methylation and histone protein modifications and they are involved in basic biological phenomena in every cell. Their role in tumorigenesis is stressed by recent unbiased large-scale studies providing evidence that several epigenetic modifiers are recurrently mutated or frequently dysregulated in multiple cancers. The interest in epigenetic marks is especially due to the fact that they are potentially reversible and thus druggable. In B-cell progenitor acute lymphoblastic leukemia (BCP-ALL) there is a relative paucity of reports on the role of histone protein modifications (acetylation, methylation, phosphorylation) as compared to acute myeloid leukemia, T-cell ALL, or other hematologic cancers, and in this setting chromatin modifications are relatively less well studied and reviewed than DNA methylation. In this paper, we discuss the biomarker associations and evidence for a driver role of dysregulated global and loci-specific histone marks, as well as mutations in epigenetic modifiers in BCP-ALL. Examples of chromatin modifiers recurrently mutated/disrupted in BCP-ALL and associated with disease outcomes include MLL1, CREBBP, NSD2, and SETD2. Altered histone marks and histone modifiers and readers may play a particular role in disease chemoresistance and relapse. We also suggest that epigenetic regulation of B-cell differentiation may have parallel roles in leukemogenesis. PMID:28054944

Electric and hybrid vehicles charge efficiency tests of ESB EV-106 lead acid batteries

Science.gov (United States)

Rowlette, J. J.

1981-01-01

Charge efficiencies were determined by measurements made under widely differing conditions of temperature, charge procedure, and battery age. The measurements were used to optimize charge procedures and to evaluate the concept of a modified, coulometric state of charge indicator. Charge efficiency determinations were made by measuring gassing rates and oxygen fractions. A novel, positive displacement gas flow meter which proved to be both simple and highly accurate is described and illustrated.

Probing the Importance of Charge Flux in Force Field Modeling.

Science.gov (United States)

Sedghamiz, Elaheh; Nagy, Balazs; Jensen, Frank

2017-08-08

We analyze the conformational dependence of atomic charges and molecular dipole moments for a selection of ∼900 conformations of peptide models of the 20 neutral amino acids. Based on a set of reference density functional theory calculations, we partition the changes into effects due to changes in bond distances, bond angles, and torsional angles and into geometry and charge flux contributions. This allows an assessment of the limitations of fixed charge force fields and indications for how to design improved force fields. The torsional degrees of freedom are the main contribution to conformational changes of atomic charges and molecular dipole moments, but indirect effects due to change in bond distances and angles account for ∼25% of the variation. Charge flux effects dominate for changes in bond distances and are also the main component of the variation in bond angles, while they are ∼25% compared to the geometry variations for torsional degrees of freedom. The geometry and charge flux contributions to some extent produce compensating effects.

Abelian scalar theory at large global charge

Energy Technology Data Exchange (ETDEWEB)

Loukas, Orestis [Albert Einstein Center for Fundamental Physics, Institute for Theoretical Physics, University of Bern (Switzerland)

2017-09-15

We elaborate on Abelian complex scalar models, which are dictated by natural actions (all couplings are of order one), at fixed and large global U(1) charge in an arbitrary number of dimensions. The ground state vertical stroke v right angle is coherently constructed by the zero modes and the appearance of a centrifugal potential is quantum mechanically verified. Using the path integral formulation we systematically analyze the quantum fluctuations around vertical stroke v right angle in order to derive an effective action for the Goldstone mode, which becomes perturbatively meaningful when the charge is large. In this regime we explicitly show, by computing the first few loop corrections, that the whole construction is stable against quantum effects, in the sense that any higher derivative couplings to Goldstone's tree-level action are suppressed by appropriate powers of the large charge. (copyright 2017 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

Criterions for fixing regulatory seismic acceleration coefficients

International Nuclear Information System (INIS)

Costes, D.

1988-03-01

Acceleration coeffficients to be taken into account in seismic areas for calculation of structures are defined in national seismic regulations. Joined to the described qualitative requirements, these coefficients represent a balance between precaution costs and avoided damages, both in terms of material repairing costs and damage to human life. Persons in charge of fixing these coefficients must be informed of corresponding quantitative aspects. Data on seismic motions occurrencies and consequences are gathered here and convoluted to mean damage evaluations. Indications on precaution costs are joined, which shows that currently recommended levels of seismic motions are high relatively to financial profitability, and represent in fact an aethical choice about human life value [fr

Energy efficient smartphone-based activity recognition using fixed-point arithmetic

OpenAIRE

Anguita, Davide; Ghio, Alessandro; Oneto, Luca; Llanas Parra, Francesc Xavier; Reyes Ortiz, Jorge Luis

2013-01-01

In this paper we propose a novel energy efficient approach for the recognition of human activities using smartphones as wearable sensing devices, targeting assisted living applications such as remote patient activity monitoring for the disabled and the elderly. The method exploits fixed-point arithmetic to propose a modified multiclass Support Vector Machine (SVM) learning algorithm, allowing to better pre- serve the smartphone battery lifetime with respect to the conventional flo...

Dipole-modified graphene with ultrahigh gas sensibility

Science.gov (United States)

Jia, Ruokun; Xie, Peng; Feng, Yancong; Chen, Zhuo; Umar, Ahmad; Wang, Yao

2018-05-01

This study reports the supramolecular assembly of functional graphene-based materials with ultrahigh gas sensing performances which are induced by charge transfer enhancement. Two typical Donor-π-Accepter (D-π-A) structure molecules 4-aminoquinoline (4AQ, μ = 3.17 Debye) and 4-hydroxyquinoline (4HQ, μ = 1.98 Debye), with different charge transfer enhancing effects, were selected to modify reduce oxide graphene (rGO) via supramolecular assembly. Notably, compared to the 4HQ-rGO, the 4AQ-rGO exhibits more significant increase of gas response (Ra/Rg = 3.79) toward 10 ppm NO2, which is ascribed to the larger dipole moment (μ) of 4AQ and hence the more intensive enhancing effect of charge transfer on the interface of rGO. Meanwhile, 4AQ-rGO sensors also reveal superior comprehensive gas sensing performances, including excellent gas sensing selectivity, linearity, repeatability and stability. It is believed that the present work demonstrates an effective supramolecular approach of modifying rGO with strong dipoles to significantly improve gas sensing properties of graphene-based materials.

Hydrophobicity of electron beam modified surface of hydroxyapatite films

Energy Technology Data Exchange (ETDEWEB)

Gregor, M., E-mail: gregor@fmph.uniba.sk [Department of Experimental Physics, Faculty of Mathematics, Physics and Informatics, Comenius University, 84248 Bratislava (Slovakia); Plecenik, T. [Department of Experimental Physics, Faculty of Mathematics, Physics and Informatics, Comenius University, 84248 Bratislava (Slovakia); Tofail, S.A.M. [Materials & Surface Science Institute, University of Limerick, Limerick (Ireland); Zahoran, M.; Truchly, M. [Department of Experimental Physics, Faculty of Mathematics, Physics and Informatics, Comenius University, 84248 Bratislava (Slovakia); Vargova, M. [Department of Inorganic Chemistry, Faculty of Natural Sciences, Comenius University, 84215 Bratislava (Slovakia); Laffir, F. [Materials & Surface Science Institute, University of Limerick, Limerick (Ireland); Plesch, G. [Department of Inorganic Chemistry, Faculty of Natural Sciences, Comenius University, 84215 Bratislava (Slovakia); Kus, P.; Plecenik, A. [Department of Experimental Physics, Faculty of Mathematics, Physics and Informatics, Comenius University, 84248 Bratislava (Slovakia)

2015-05-15

Highlights: • Surface potential of hydroxyapatite films were modified by focused electron beam. • Micron-sized domains of modified surface potential were created. • Wettability and surface free energy of the irradiated areas was studied. • Possible mechanisms of increased surface hydrophobicity are discussed. - Abstract: Arrays of micron-sized domains of modified surface potential were created on hydroxyapatite films by mid-energy (20 keV) electron beam irradiation available in a laboratory scanning electron microscope. The dosage of electron beam was varied between 10{sup −3} and 10{sup 3} μC/cm{sup 2} to inject charge into the film surface. Contrary to the conventional electrowetting theory, the dosage of injected charge used in creating such microdomains caused a gradual increase of the water contact angle from 57° to 93° due to the elimination of the polar component of the surface free energy. Surface contamination by carbonaceous species can be held only partially responsible for such behavior at lower dosage of electron beam. A transfer of free surface charge to water and an electron beam induced disruption of polar orientation of OH ions have been attributed to be influencial factors in the overall dewetting behavior.

Chitosan scaffold modified with D-(+) raffinose and enriched with thiol-modified gelatin for improved osteoblast adhesion

International Nuclear Information System (INIS)

Galli, C; Parisi, L; Smerieri, A; Lumetti, S; Manfredi, E; Macaluso, G M; Elviri, L; Bianchera, A; Bettini, R; Lagonegro, P

2016-01-01

The aim of the present study was to investigate whether chitosan-based scaffolds modified with D-(+) raffinose and enriched with thiol-modified gelatin could selectively improve osteoblast adhesion and proliferation. 2, 3 and 4.5% chitosan films were prepared. Chitosan suitability for tissue engineering was confirmed by protein adsorption assay. Scaffolds were incubated with a 2.5 mg ml −1 BSA solution and the decrease of protein content in the supernatants was measured by spectrophotometry. Chitosan films were then enriched with thiol-modified gelatin and their ability to bind BSA was also measured. Then, 2% chitosan discs with or without thiol-modified gelatin were used as culture substrates for MC3T3-E1 cells. After 72 h cells were stained with trypan blue or with calcein AM and propidium iodide for morphology, viability and proliferation assays. Moreover, cell viability was measured at 48, 72, 96 and 168 h to obtain a growth curve. Chitosan films efficiently bound and retained BSA proportionally to the concentration of chitosan discs. The amount of protein retained was higher on chitosan enriched with thiol-modified gelatin. Moreover, chitosan discs allowed the adhesion and the viability of cells, but inhibited their proliferation. The functionalization of chitosan with thiol-modified gelatin enhanced cell spreading and proliferation. Our data confirm that chitosan is a suitable material for tissue engineering. Moreover, our data show that the enrichment of chitosan with thiol-modified gelatin enhances its biological properties. (paper)

Cyanobacterial flv4-2 Operon-Encoded Proteins Optimize Light Harvesting and Charge Separation in Photosystem II.

Science.gov (United States)

Chukhutsina, Volha; Bersanini, Luca; Aro, Eva-Mari; van Amerongen, Herbert

2015-05-01

Photosystem II (PSII) complexes drive the water-splitting reaction necessary to transform sunlight into chemical energy. However, too much light can damage and disrupt PSII. In cyanobacteria, the flv4-2 operon encodes three proteins (Flv2, Flv4, and Sll0218), which safeguard PSII activity under air-level CO2 and in high light conditions. However, the exact mechanism of action of these proteins has not been clarified yet. We demonstrate that the PSII electron transfer properties are influenced by the flv4-2 operon-encoded proteins. Accelerated secondary charge separation kinetics was observed upon expression/overexpression of the flv4-2 operon. This is likely induced by docking of the Flv2/Flv4 heterodimer in the vicinity of the QB pocket of PSII, which, in turn, increases the QB redox potential and consequently stabilizes forward electron transfer. The alternative electron transfer route constituted by Flv2/Flv4 sequesters electrons from QB(-) guaranteeing the dissipation of excess excitation energy in PSII under stressful conditions. In addition, we demonstrate that in the absence of the flv4-2 operon-encoded proteins, about 20% of the phycobilisome antenna becomes detached from the reaction centers, thus decreasing light harvesting. Phycobilisome detachment is a consequence of a decreased relative content of PSII dimers, a feature observed in the absence of the Sll0218 protein. Copyright © 2015 The Author. Published by Elsevier Inc. All rights reserved.

Charge collection in the Silicon Drift Detectors of the ALICE experiment

CERN Document Server

Alessandro, B; Batigne, G; Beolé, S; Biolcati, E; Cerello, P; Coli, S; Corrales Morales, Y; Crescio, E; De Remigis, P; Falchieri, D; Giraudo, G; Giubellino, P; Lea, R; Marzari Chiesa, A; Masera, M; Mazza, G; Ortona, G; Prino, F; Ramello, L; Rashevsky, A; Riccati, L; Rivetti, A; Senyukov, S; Siciliano, M; Sitta, M; Subieta, M; Toscano, L; Tosello, F

2010-01-01

A detailed study of charge collection efficiency has been performed on the Silicon Drift Detectors (SDD) of the ALICE experiment. Three different methods to study the collected charge as a function of the drift time have been implemented. The first approach consists in measuring the charge at different injection distances moving an infrared laser by means of micrometric step motors. The second method is based on the measurement of the charge injected by the laser at fixed drift distance and varying the drift field, thus changing the drift time. In the last method, the measurement of the charge deposited by atmospheric muons is used to study the charge collection efficiency as a function of the drift time. The three methods gave consistent results and indicated that no charge loss during the drift is observed for the sensor types used in 99% of the SDD modules mounted on the ALICE Inner Tracking System. The atmospheric muons have also been used to test the effect of the zero-suppression applied to reduce the d...

Damage induced by high energy multiply charged oxygen ions in oxide coated silicon

Energy Technology Data Exchange (ETDEWEB)

Dhole, S.D. [Department of Physics, University of Pune, Pune 411 007 (India)]. E-mail: sanjay@physics.unipune.ernet.in; Dahiwale, S.S. [Department of Physics, University of Pune, Pune 411 007 (India); Kulkarni, V.R. [Department of Physics, University of Pune, Pune 411 007 (India); Bogle, K.A. [Department of Physics, University of Pune, Pune 411 007 (India); Shinde, N.S. [Ecotopia Science Institute, Division of Energy Science, Nagoya University, Nagoya (Japan); Bhoraskar, V.N. [Department of Physics, University of Pune, Pune 411 007 (India)

2006-03-15

P-type oxide coated silicon samples of resistivity 120 {omega} cm were irradiated with 60 MeV oxygen ions of fixed charge states 4{sup +}, 5{sup +}, 6{sup +} and 7{sup +} at an equal fluence of, {phi}, {approx}10{sup 13} ions/cm{sup 2}. The induced damage was estimated by Hall voltage, Hall coefficient, carrier concentration and lifetime of minority carriers. The results indicate that Hall voltage (V {sub H}) and Hall coefficient (R {sub H}) increases, while carrier concentration (n) decreases with the charge state of impinging oxygen ions. The V {sub H} increases from 22 mV to 76.5 mV at typical current of 0.5 mA, R {sub H} from 0.42 x 10{sup 5} cm{sup 3}/C to 2.16 x 10{sup 5} cm{sup 3}/C and n decreases from 9 x 10{sup 13} cm{sup -3} to 2.88 x 10{sup 13} cm{sup -3} for the different charge states. This fact is an evidence that the oxygen ions with an individual fixed charge state passing through very thin 40 A layer of silicon dioxide, induces significant damage at the SiO{sub 2}-Si interface through the mechanism of electronic stopping power. The lifetime of minority charge carriers, {tau} (bulk property), remains constant at around 6 {mu}s for all the charge states of the 60 MeV energy oxygen ion irradiated samples at a constant fluence of, {phi}, 10{sup 13} ions/cm{sup 2}.

Role of molecular charge in nucleocytoplasmic transport.

Directory of Open Access Journals (Sweden)

Alexander Goryaynov

Full Text Available Transport of genetic materials and proteins between the nucleus and cytoplasm of eukaryotic cells is mediated by nuclear pore complexes (NPCs. A selective barrier formed by phenylalanine-glycine (FG nucleoporins (Nups with net positive charges in the NPC allows for passive diffusion of signal-independent small molecules and transport-receptor facilitated translocation of signal-dependent cargo molecules. Recently, negative surface charge was postulated to be another essential criterion for selective passage through the NPC. However, the charge-driven mechanism in determining the transport kinetics and spatial transport route for either passive diffusion or facilitated translocation remains obscure. Here we employed high-speed single-molecule fluorescence microscopy with an unprecedented spatiotemporal resolution of 9 nm and 400 µs to uncover these mechanistic fundamentals for nuclear transport of charged substrates through native NPCs. We found that electrostatic interaction between negative surface charges on transiting molecules and the positively charged FG Nups, although enhancing their probability of binding to the NPC, never plays a dominant role in determining their nuclear transport mode or spatial transport route. A 3D reconstruction of transport routes revealed that small signal-dependent endogenous cargo protein constructs with high positive surface charges that are destined to the nucleus, rather than repelled from the NPC as suggested in previous models, passively diffused through an axial central channel of the NPC in the absence of transport receptors. Finally, we postulated a comprehensive map of interactions between transiting molecules and FG Nups during nucleocytoplasmic transport by combining the effects of molecular size, signal and surface charge.

Electrical charging effects on the sliding friction of a model nano-confined ionic liquid

Energy Technology Data Exchange (ETDEWEB)

Capozza, R.; Vanossi, A. [International School for Advanced Studies (SISSA), Via Bonomea 265, 34136 Trieste (Italy); CNR-IOM Democritos National Simulation Center, Via Bonomea 265, 34136 Trieste (Italy); Benassi, A. [CNR-IOM Democritos National Simulation Center, Via Bonomea 265, 34136 Trieste (Italy); Institute for Materials Science and Max Bergmann Center of Biomaterials, TU Dresden, 01062 Dresden (Germany); Tosatti, E. [International School for Advanced Studies (SISSA), Via Bonomea 265, 34136 Trieste (Italy); CNR-IOM Democritos National Simulation Center, Via Bonomea 265, 34136 Trieste (Italy); International Centre for Theoretical Physics (ICTP), Strada Costiera 11, 34014 Trieste (Italy)

2015-10-14

Recent measurements suggest the possibility to exploit ionic liquids (ILs) as smart lubricants for nano-contacts, tuning their tribological and rheological properties by charging the sliding interfaces. Following our earlier theoretical study of charging effects on nanoscale confinement and squeezout of a model IL, we present here molecular dynamics simulations of the frictional and lubrication properties of that model under charging conditions. First, we describe the case when two equally charged plates slide while being held together to a confinement distance of a few molecular layers. The shear sliding stress is found to rise strongly and discontinuously as the number of IL layers decreases stepwise. However, the shear stress shows, within each given number of layers, only a weak dependence upon the precise value of the normal load, a result in agreement with data extracted from recent experiments. We subsequently describe the case of opposite charging of the sliding plates and follow the shear stress when the charging is slowly and adiabatically reversed in the course of time, under fixed load. Despite the fixed load, the number and structure of the confined IL layers change with changing charge, and that in turn drives strong friction variations. The latter involves first of all charging-induced freezing of the IL film, followed by a discharging-induced melting, both made possible by the nanoscale confinement. Another mechanism for charging-induced frictional changes is a shift of the plane of maximum shear from mid-film to the plate-film interface, and vice versa. While these occurrences and results invariably depend upon the parameters of the model IL and upon its specific interaction with the plates, the present study helps identifying a variety of possible behavior, obtained under very simple assumptions, while connecting it to an underlying equilibrium thermodynamics picture.

Further along the Road Less Traveled: AMBER ff15ipq, an Original Protein Force Field Built on a Self-Consistent Physical Model

Science.gov (United States)

2016-01-01

We present the AMBER ff15ipq force field for proteins, the second-generation force field developed using the Implicitly Polarized Q (IPolQ) scheme for deriving implicitly polarized atomic charges in the presence of explicit solvent. The ff15ipq force field is a complete rederivation including more than 300 unique atomic charges, 900 unique torsion terms, 60 new angle parameters, and new atomic radii for polar hydrogens. The atomic charges were derived in the context of the SPC/Eb water model, which yields more-accurate rotational diffusion of proteins and enables direct calculation of nuclear magnetic resonance (NMR) relaxation parameters from molecular dynamics simulations. The atomic radii improve the accuracy of modeling salt bridge interactions relative to contemporary fixed-charge force fields, rectifying a limitation of ff14ipq that resulted from its use of pair-specific Lennard-Jones radii. In addition, ff15ipq reproduces penta-alanine J-coupling constants exceptionally well, gives reasonable agreement with NMR relaxation rates, and maintains the expected conformational propensities of structured proteins/peptides, as well as disordered peptides—all on the microsecond (μs) time scale, which is a critical regime for drug design applications. These encouraging results demonstrate the power and robustness of our automated methods for deriving new force fields. All parameters described here and the mdgx program used to fit them are included in the AmberTools16 distribution. PMID:27399642

Electric Vehicle Smart Charging using Dynamic Price Signal

DEFF Research Database (Denmark)

Martinenas, Sergejus; Pedersen, Anders Bro; Marinelli, Mattia

2014-01-01

, however, be resolved by using intelligent EV charging strategies, commonly referred to as ”Smart Charging”. The basic approach involves modifying the default vehicle charging scheme of ”immediate charging”, to a more optimal one that is derived from insight into the current state of the grid. This work......With yearly increases in Electric Vehicle (EV) sales, the future for electric mobility continues to brighten, and with more vehicles hitting the roads every day, the energy requirements on the grid will increase, potentially causing low-voltage distribution grid congestion. This problem can...... proposed in this paper, involves a real-time control strategy for charging the EV using a dynamic price tariff, with the objective of minimizing the charging cost. Two different charging scenario are investigated, and the results are verified by experiments on a real Electric Vehicle. Finally, the costs...

Search for Doubly-Charged Higgs Bosons at LEP

CERN Document Server

Achard, P.; Aguilar-Benitez, M.; Alcaraz, J.; Alemanni, G.; Allaby, J.; Aloisio, A.; Alviggi, M.G.; Anderhub, H.; Andreev, Valery P.; Anselmo, F.; Arefev, A.; Azemoon, T.; Aziz, T.; Bagnaia, P.; Bajo, A.; Baksay, G.; Baksay, L.; Baldew, S.V.; Banerjee, S.; Barczyk, A.; Barillere, R.; Bartalini, P.; Basile, M.; Batalova, N.; Battiston, R.; Bay, A.; Becattini, F.; Becker, U.; Behner, F.; Bellucci, L.; Berbeco, R.; Berdugo, J.; Berges, P.; Bertucci, B.; Betev, B.L.; Biasini, M.; Biglietti, M.; Biland, A.; Blaising, J.J.; Blyth, S.C.; Bobbink, G.J.; Bohm, A.; Boldizsar, L.; Borgia, B.; Bottai, S.; Bourilkov, D.; Bourquin, M.; Braccini, S.; Branson, J.G.; Brochu, F.; Burger, J.D.; Burger, W.J.; Cai, X.D.; Capell, M.; Cara Romeo, G.; Carlino, G.; Cartacci, A.; Casaus, J.; Cavallari, F.; Cavallo, N.; Cecchi, C.; Cerrada, M.; Chamizo, M.; Chang, Y.H.; Chemarin, M.; Chen, A.; Chen, G.; Chen, G.M.; Chen, H.F.; Chen, H.S.; Chiefari, G.; Cifarelli, L.; Cindolo, F.; Clare, I.; Clare, R.; Coignet, G.; Colino, N.; Costantini, S.; de la Cruz, B.; Cucciarelli, S.; van Dalen, J.A.; de Asmundis, R.; Deglon, P.; Debreczeni, J.; Degre, A.; Dehmelt, K.; Deiters, K.; della Volpe, D.; Delmeire, E.; Denes, P.; DeNotaristefani, F.; De Salvo, A.; Diemoz, M.; Dierckxsens, M.; Dionisi, C.; Dittmar, M.; Doria, A.; Dova, M.T.; Duchesneau, D.; Duda, M.; Echenard, B.; Eline, A.; El Hage, A.; El Mamouni, H.; Engler, A.; Eppling, F.J.; Extermann, P.; Falagan, M.A.; Falciano, S.; Favara, A.; Fay, J.; Fedin, O.; Felcini, M.; Ferguson, T.; Fesefeldt, H.; Fiandrini, E.; Field, J.H.; Filthaut, F.; Fisher, P.H.; Fisher, W.; Fisk, I.; Forconi, G.; Freudenreich, K.; Furetta, C.; Galaktionov, Iouri; Ganguli, S.N.; Garcia-Abia, Pablo; Gataullin, M.; Gentile, S.; Giagu, S.; Gong, Z.F.; Grenier, Gerald Jean; Grimm, O.; Gruenewald, M.W.; Guida, M.; van Gulik, R.; Gupta, V.K.; Gurtu, A.; Gutay, L.J.; Haas, D.; Hatzifotiadou, D.; Hebbeker, T.; Herve, Alain; Hirschfelder, J.; Hofer, H.; Hohlmann, M.; Holzner, G.; Hou, S.R.; Hu, Y.; Jin, B.N.; Jones, Lawrence W.; de Jong, P.; Josa-Mutuberria, I.; Kafer, D.; Kaur, M.; Kienzle-Focacci, M.N.; Kim, J.K.; Kirkby, Jasper; Kittel, W.; Klimentov, A.; Konig, A.C.; Kopal, M.; Koutsenko, V.; Kraber, M.; Kraemer, R.W.; Kruger, A.; Kunin, A.; Ladron de Guevara, P.; Laktineh, I.; Landi, G.; Lebeau, M.; Lebedev, A.; Lebrun, P.; Lecomte, P.; Lecoq, P.; Le Coultre, P.; Le Goff, J.M.; Leiste, R.; Levtchenko, M.; Levtchenko, P.; Li, C.; Likhoded, S.; Lin, C.H.; Lin, W.T.; Linde, F.L.; Lista, L.; Liu, Z.A.; Lohmann, W.; Longo, E.; Lu, Y.S.; Luci, C.; Luminari, L.; Lustermann, W.; Ma, W.G.; Malgeri, L.; Malinin, A.; Mana, C.; Mans, J.; Martin, J.P.; Marzano, F.; Mazumdar, K.; McNeil, R.R.; Mele, S.; Merola, L.; Meschini, M.; Metzger, W.J.; Mihul, A.; Milcent, H.; Mirabelli, G.; Mnich, J.; Mohanty, G.B.; Muanza, G.S.; Muijs, A.J.M.; Musicar, B.; Musy, M.; Nagy, S.; Natale, S.; Napolitano, M.; Nessi-Tedaldi, F.; Newman, H.; Nisati, A.; Novak, T.; Kluge, Hannelies; Ofierzynski, R.; Organtini, G.; Pal, I.; Palomares, C.; Paolucci, P.; Paramatti, R.; Passaleva, G.; Patricelli, S.; Paul, Thomas Cantzon; Pauluzzi, M.; Paus, C.; Pauss, F.; Pedace, M.; Pensotti, S.; Perret-Gallix, D.; Petersen, B.; Piccolo, D.; Pierella, F.; Pioppi, M.; Piroue, P.A.; Pistolesi, E.; Plyaskin, V.; Pohl, M.; Pojidaev, V.; Pothier, J.; Prokofev, D.; Quartieri, J.; Rahal-Callot, G.; Rahaman, Mohammad Azizur; Raics, P.; Raja, N.; Ramelli, R.; Rancoita, P.G.; Ranieri, R.; Raspereza, A.; Razis, P.; Ren, D.; Rescigno, M.; Reucroft, S.; Riemann, S.; Riles, Keith; Roe, B.P.; Romero, L.; Rosca, A.; Rosier-Lees, S.; Roth, Stefan; Rosenbleck, C.; Roux, B.; Rubio, J.A.; Ruggiero, G.; Rykaczewski, H.; Sakharov, A.; Saremi, S.; Sarkar, S.; Salicio, J.; Sanchez, E.; Schafer, C.; Schegelsky, V.; Schopper, H.; Schotanus, D.J.; Sciacca, C.; Servoli, L.; Shevchenko, S.; Shivarov, N.; Shoutko, V.; Shumilov, E.; Shvorob, A.; Son, D.; Souga, C.; Spillantini, P.; Steuer, M.; Stickland, D.P.; Stoyanov, B.; Straessner, A.; Sudhakar, K.; Sultanov, G.; Sun, L.Z.; Sushkov, S.; Suter, H.; Swain, J.D.; Szillasi, Z.; Tang, X.W.; Tarjan, P.; Tauscher, L.; Taylor, L.; Tellili, B.; Teyssier, D.; Timmermans, Charles; Ting, Samuel C.C.; Ting, S.M.; Tonwar, S.C.; Toth, J.; Tully, C.; Tung, K.L.; Ulbricht, J.; Valente, E.; Van de Walle, R.T.; Vasquez, R.; Veszpremi, V.; Vesztergombi, G.; Vetlitsky, I.; Vicinanza, D.; Viertel, G.; Villa, S.; Vivargent, M.; Vlachos, S.; Vodopianov, I.; Vogel, H.; Vogt, H.; Vorobev, I.; Vorobyov, A.A.; Wadhwa, M.; Wang, Q.; Wang, X.L.; Wang, Z.M.; Weber, M.; Wienemann, P.; Wilkens, H.; Wynhoff, S.; Xia, L.; Xu, Z.Z.; Yamamoto, J.; Yang, B.Z.; Yang, C.G.; Yang, H.J.; Yang, M.; Yeh, S.C.; Zalite, A.; Zalite, Yu.; Zhang, Z.P.; Zhao, J.; Zhu, G.Y.; Zhu, R.Y.; Zhuang, H.L.; Zichichi, A.; Zimmermann, B.; Zoller, M.

2003-01-01

Doubly-charged Higgs bosons are searched for in e^+e^- collision data collected with the L3 detector at LEP at centre-of-mass energies up to 209 GeV. Final states with four leptons are analysed to tag the pair-production of doubly charged Higgs bosons. No significant excess is found and lower limits at 95% confidence level on the doubly-charged Higgs boson mass are derived. They vary from 95.5 GeV to 100.2 GeV, depending on the decay mode. Doubly-charged Higgs bosons which couple to electrons would modify the cross section and forward-backward asymmetry of the e^+e^- -> e^+e^- process. The measurements of these quantities do not deviate from the Standard Model expectations and doubly-charged Higgs bosons with masses up to the order of a TeV are excluded.

Modifiers of notch transcriptional activity identified by genome-wide RNAi

Directory of Open Access Journals (Sweden)

Firnhaber Christopher B

2010-10-01

Full Text Available Abstract Background The Notch signaling pathway regulates a diverse array of developmental processes, and aberrant Notch signaling can lead to diseases, including cancer. To obtain a more comprehensive understanding of the genetic network that integrates into Notch signaling, we performed a genome-wide RNAi screen in Drosophila cell culture to identify genes that modify Notch-dependent transcription. Results Employing complementary data analyses, we found 399 putative modifiers: 189 promoting and 210 antagonizing Notch activated transcription. These modifiers included several known Notch interactors, validating the robustness of the assay. Many novel modifiers were also identified, covering a range of cellular localizations from the extracellular matrix to the nucleus, as well as a large number of proteins with unknown function. Chromatin-modifying proteins represent a major class of genes identified, including histone deacetylase and demethylase complex components and other chromatin modifying, remodeling and replacement factors. A protein-protein interaction map of the Notch-dependent transcription modifiers revealed that a large number of the identified proteins interact physically with these core chromatin components. Conclusions The genome-wide RNAi screen identified many genes that can modulate Notch transcriptional output. A protein interaction map of the identified genes highlighted a network of chromatin-modifying enzymes and remodelers that regulate Notch transcription. Our results open new avenues to explore the mechanisms of Notch signal regulation and the integration of this pathway into diverse cellular processes.

Modified Folin-Ciocalteu antioxidant capacity assay for measuring lipophilic antioxidants.

Science.gov (United States)

Berker, Kadriye Isil; Ozdemir Olgun, F Ayca; Ozyurt, Dilek; Demirata, Birsen; Apak, Resat

2013-05-22

The Folin-Ciocalteu (FC) method of performing a total phenolics assay, originally developed for protein determination, has recently evolved as a total antioxidant capacity assay but was found to be incapable of measuring lipophilic antioxidants due to the high affinity of the FC chromophore, that is, multivalent-charged phospho-tungsto-molybdate(V), toward water. Thus, the FC method was modified and standardized so as to enable simultaneous measurement of lipophilic and hydrophilic antioxidants in NaOH-added isobutanol-water medium. Optimal conditions were as follows: dilution ratio of aqueous FC reagent with iso-BuOH (1:2, v/v), final NaOH concentration of 3.5 × 10(-2) M, reaction time of 20 min, and maximum absorption wavelength of 665 nm. The modified procedure was successfully applied to the total antioxidant capacity assay of trolox, quercetin, ascorbic acid, gallic acid, catechin, caffeic acid, ferulic acid, rosmarinic acid, glutathione, and cysteine, as well as of lipophilic antioxidants such as α-tocopherol (vitamin E), butylated hydroxyanisole, butylated hydroxytoluene, tertiary butylhydroquinone, lauryl gallate, and β-carotene. The modified FC method reliably quantified ascorbic acid, whereas the conventional method could not. The modified method was reproducible and additive in terms of total antioxidant capacity values of constituents of complex mixtures such as olive oil extract and herbal tea infusion. The trolox equivalent antioxidant capacities of the tested antioxidant compounds correlated well with those found by the Cupric Reducing Antioxidant Capacity reference method.

The charge form factor, the quadrupole moment and the photodisintegration of 6Li

International Nuclear Information System (INIS)

Susila, S.; Srinivasa Rao, K.

1981-01-01

The root mean square radius, the charge form factor, the charge density, the quadrupole moment and the bremsstrahlung weighted cross section for the photodisintegration of 6 Li, are calculated using a polarised cluster model wave function, which is modified to take into account, in its relative motion part, the requirement of a shell model node. A set of parameters, in the modified cluster model wave function, which account for the available experimental data for the afore-said quantities, is determined. (author)

A modified space charge routine for high intensity bunched beams

International Nuclear Information System (INIS)

Lapostolle, P.; Garnett, R.W.; Wangler, T.P.

1996-01-01

In 1991 a space charge calculation for bunched beam with a three-dimensional ellipsoid was proposed, replacing the usual SCHEFF routines. It removes the cylindrical symmetry required in SCHEFF and avoids the point to point interaction computation, whose number of simulation points is limited. This routine has now been improved with the introduction of two or three ellipsoids giving a good representation of the complex non-symmetrical form of the bunch (unlike the 3-d ellipsoidal assumption). The ellipsoidal density distributions are computed with a new method, avoiding the difficulty encountered near the centre (the axis in 2-d problems) by the previous method. It also provides a check of the ellipsoidal symmetry for each part of the distribution. Finally, the Fourier analysis reported in 1991 has been replaced by a very convenient Hermite expansion, which gives a simple but accurate representation of practical distributions. Comparisons with other space charge routines have been made, particularly with the ones applying other techniques such as SCHEFF. Introduced in the versatile beam dynamics code DYNAC, it should provide a good tool for the study of the various parameters responsible for the halo formation in high intensity linacs. (orig.)

What Protein Charging (and Supercharging) Reveal about the Mechanism of Electrospray Ionization

Science.gov (United States)

Ogorzalek Loo, Rachel R.; Lakshmanan, Rajeswari; Loo, Joseph A.

2014-10-01

Understanding the charging mechanism of electrospray ionization is central to overcoming shortcomings such as ion suppression or limited dynamic range, and explaining phenomena such as supercharging. Towards that end, we explore what accumulated observations reveal about the mechanism of electrospray. We introduce the idea of an intermediate region for electrospray ionization (and other ionization methods) to account for the facts that solution charge state distributions (CSDs) do not correlate with those observed by ESI-MS (the latter bear more charge) and that gas phase reactions can reduce, but not increase, the extent of charging. This region incorporates properties (e.g., basicities) intermediate between solution and gas phase. Assuming that droplet species polarize within the high electric field leads to equations describing ion emission resembling those from the equilibrium partitioning model. The equations predict many trends successfully, including CSD shifts to higher m/z for concentrated analytes and shifts to lower m/z for sprays employing smaller emitter opening diameters. From this view, a single mechanism can be formulated to explain how reagents that promote analyte charging ("supercharging") such as m-NBA, sulfolane, and 3-nitrobenzonitrile increase analyte charge from "denaturing" and "native" solvent systems. It is suggested that additives' Brønsted basicities are inversely correlated to their ability to shift CSDs to lower m/z in positive ESI, as are Brønsted acidities for negative ESI. Because supercharging agents reduce an analyte's solution ionization, excess spray charge is bestowed on evaporating ions carrying fewer opposing charges. Brønsted basicity (or acidity) determines how much ESI charge is lost to the agent (unavailable to evaporating analyte).

Robust fixed-time synchronization for uncertain complex-valued neural networks with discontinuous activation functions.

Science.gov (United States)

Ding, Xiaoshuai; Cao, Jinde; Alsaedi, Ahmed; Alsaadi, Fuad E; Hayat, Tasawar

2017-06-01

This paper is concerned with the fixed-time synchronization for a class of complex-valued neural networks in the presence of discontinuous activation functions and parameter uncertainties. Fixed-time synchronization not only claims that the considered master-slave system realizes synchronization within a finite time segment, but also requires a uniform upper bound for such time intervals for all initial synchronization errors. To accomplish the target of fixed-time synchronization, a novel feedback control procedure is designed for the slave neural networks. By means of the Filippov discontinuity theories and Lyapunov stability theories, some sufficient conditions are established for the selection of control parameters to guarantee synchronization within a fixed time, while an upper bound of the settling time is acquired as well, which allows to be modulated to predefined values independently on initial conditions. Additionally, criteria of modified controller for assurance of fixed-time anti-synchronization are also derived for the same system. An example is included to illustrate the proposed methodologies. Copyright © 2017 Elsevier Ltd. All rights reserved.

On the classical dynamics of charges in non-commutative QED

International Nuclear Information System (INIS)

Fatollahi, A.H.; Mohammadzadeh, H.

2004-01-01

Following Wong's approach to formulating the classical dynamics of charged particles in non-Abelian gauge theories, we derive the classical equations of motion of a charged particle in U(1) gauge theory on non-commutative space, the so-called non-commutative QED. In the present use of the procedure, it is observed that the definition of the mechanical momenta should be modified. The derived equations of motion manifest the previous statement about the dipole behavior of the charges in non-commutative space. (orig.)

Charged particle-like branes in ABJM

CERN Document Server

Gutierrez, Norberto; Rodriguez-Gomez, Diego

2010-01-01

We study the effect of adding lower dimensional brane charges to the 't Hooft monopole, di-baryon and baryon vertex configurations in $AdS_4 \\times \\mathbb{P}^3$. We show that these configurations capture the background fluxes in a way that depends on the induced charges, and therefore, require additional fundamental strings in order to cancel the worldvolume tadpoles. The study of the dynamics reveals that the charges must lie inside some interval in order to find well defined configurations, a situation familiar from the baryon vertex in $AdS_5 \\times S^5$ with charges. For the baryon vertex and the di-baryon the number of fundamental strings must also lie inside an allowed interval. Our configurations are sensitive to the flat $B$-field recently suggested in the literature. We make some comments on its possible role. We also discuss how these configurations are modified in the presence of a non-zero Romans mass.

Critical phenomena of static charged AdS black holes in conformal gravity

Directory of Open Access Journals (Sweden)

Wei Xu

2014-09-01

Full Text Available The extended thermodynamics of static charged AdS black holes in conformal gravity is analyzed. The P–V criticality of these black holes has some unusual features. There exists a single critical point with critical temperature Tc and critical pressure Pc. At fixed T>Tc (or at fixed P>Pc, there are two zeroth order phase transition points but no first order phase transition points. The systems favors large pressure states at constant T, or high temperature states at constant P.

Intein-modified enzymes, their production and industrial applications

Science.gov (United States)

Apgar, James; Lessard, Philip; Raab, Michael R.; Shen, Binzhang; Lazar, Gabor; de la Vega, Humberto

2016-10-11

A method of predicting an intein insertion site in a protein that will lead to a switching phenotype is provided. The method includes identifying a plurality of C/T/S sites within the protein; selecting from the plurality of C/T/S/ sites those that are ranked 0.75 or higher by a support vector machine, within ten angstroms of the active site of the protein, and at or near a loop-.beta.-sheet junction or a loop-.alpha.-helix junction. A method of controlling protein activity and hosts including proteins with controlled activity are also provided. Also, intein modified proteins and plants containing intein modified proteins are provided.

Protein-induced geometric constraints and charge transfer in bacteriochlorophyll-histidine complexes in LH2.

Science.gov (United States)

Wawrzyniak, Piotr K; Alia, A; Schaap, Roland G; Heemskerk, Mattijs M; de Groot, Huub J M; Buda, Francesco

2008-12-14

Bacteriochlorophyll-histidine complexes are ubiquitous in nature and are essential structural motifs supporting the conversion of solar energy into chemically useful compounds in a wide range of photosynthesis processes. A systematic density functional theory study of the NMR chemical shifts for histidine and for bacteriochlorophyll-a-histidine complexes in the light-harvesting complex II (LH2) is performed using the BLYP functional in combination with the 6-311++G(d,p) basis set. The computed chemical shift patterns are consistent with available experimental data for positive and neutral(tau) (N(tau) protonated) crystalline histidines. The results for the bacteriochlorophyll-a-histidine complexes in LH2 provide evidence that the protein environment is stabilizing the histidine close to the Mg ion, thereby inducing a large charge transfer of approximately 0.5 electronic equivalent. Due to this protein-induced geometric constraint, the Mg-coordinated histidine in LH2 appears to be in a frustrated state very different from the formal neutral(pi) (N(pi) protonated) form. This finding could be important for the understanding of basic functional mechanisms involved in tuning the electronic properties and exciton coupling in LH2.

Discretization of the induced-charge boundary integral equation.

Science.gov (United States)

Bardhan, Jaydeep P; Eisenberg, Robert S; Gillespie, Dirk

2009-07-01

Boundary-element methods (BEMs) for solving integral equations numerically have been used in many fields to compute the induced charges at dielectric boundaries. In this paper, we consider a more accurate implementation of BEM in the context of ions in aqueous solution near proteins, but our results are applicable more generally. The ions that modulate protein function are often within a few angstroms of the protein, which leads to the significant accumulation of polarization charge at the protein-solvent interface. Computing the induced charge accurately and quickly poses a numerical challenge in solving a popular integral equation using BEM. In particular, the accuracy of simulations can depend strongly on seemingly minor details of how the entries of the BEM matrix are calculated. We demonstrate that when the dielectric interface is discretized into flat tiles, the qualocation method of Tausch [IEEE Trans Comput.-Comput.-Aided Des. 20, 1398 (2001)] to compute the BEM matrix elements is always more accurate than the traditional centroid-collocation method. Qualocation is not more expensive to implement than collocation and can save significant computational time by reducing the number of boundary elements needed to discretize the dielectric interfaces.

Discretization of the induced-charge boundary integral equation.

Energy Technology Data Exchange (ETDEWEB)

Bardhan, J. P.; Eisenberg, R. S.; Gillespie, D.; Rush Univ. Medical Center

2009-07-01

Boundary-element methods (BEMs) for solving integral equations numerically have been used in many fields to compute the induced charges at dielectric boundaries. In this paper, we consider a more accurate implementation of BEM in the context of ions in aqueous solution near proteins, but our results are applicable more generally. The ions that modulate protein function are often within a few angstroms of the protein, which leads to the significant accumulation of polarization charge at the protein-solvent interface. Computing the induced charge accurately and quickly poses a numerical challenge in solving a popular integral equation using BEM. In particular, the accuracy of simulations can depend strongly on seemingly minor details of how the entries of the BEM matrix are calculated. We demonstrate that when the dielectric interface is discretized into flat tiles, the qualocation method of Tausch et al. [IEEE Trans Comput.-Comput.-Aided Des. 20, 1398 (2001)] to compute the BEM matrix elements is always more accurate than the traditional centroid-collocation method. Qualocation is not more expensive to implement than collocation and can save significant computational time by reducing the number of boundary elements needed to discretize the dielectric interfaces.

Ethylene glycol assisted preparation of Ti(4+)-modified polydopamine coated magnetic particles with rough surface for capture of phosphorylated proteins.

Science.gov (United States)

Ma, Xiangdong; Ding, Chun; Yao, Xin; Jia, Li

2016-07-27

The reversible protein phosphorylation is very important in regulating almost all aspects of cell life, while the enrichment of phosphorylated proteins still remains a technical challenge. In this work, polydopamine (PDA) modified magnetic particles with rough surface (rPDA@Fe3O4) were synthesized by introduction of ethylene glycol in aqueous solution. The PDA coating possessing a wealth of catechol hydroxyl groups could serve as an active medium to immobilize titanium ions through the metal-catechol chelation, which makes the fabrication of titanium ions modified rPDA@Fe3O4 particles (Ti(4+)-rPDA@Fe3O4) simple and very convenient. The spherical Ti(4+)-rPDA@Fe3O4 particles have a surface area of 37.7 m(2) g(-1) and superparamagnetism with a saturation magnetization value of 38.4 emu g(-1). The amount of Ti element in the particle was measured to be 3.93%. And the particles demonstrated good water dispersibility. The particles were used as adsorbents for capture of phosphorylated proteins and they demonstrated affinity and specificity for phosphorylated proteins due to the specific binding sites (Ti(4+)). Factors affecting the adsorption of phosphorylated proteins on Ti(4+)-rPDA@Fe3O4 particles were investigated. The adsorption capacity of Ti(4+)-rPDA@Fe3O4 particles for κ-casein was 1105.6 mg g(-1). Furthermore, the particles were successfully applied to isolate phosphorylated proteins in milk samples, which demonstrated that Ti(4+)-rPDA@Fe3O4 particles had potential application in selective separation of phosphorylated proteins. Copyright © 2016 Elsevier B.V. All rights reserved.

Equivalent circuit modeling of space charge dominated magnetically insulated transmission lines

Energy Technology Data Exchange (ETDEWEB)

Hiraoka, Kazuki; Nakajima, Mitsuo; Horioka, Kazuhiko

1997-12-31

A new equivalent circuit model for space charge dominated MITLs (Magnetically Insulated Transmission Lines) was developed. MITLs under high power operation are dominated with space charge current flowing between anode and cathode. Conventional equivalent circuit model does not account for space charge effects on power flow. The model was modified to discuss the power transportation through the high power MITLs. With this model, it is possible to estimate the effects of space charge current on the power flow efficiency, without using complicated particle code simulations. (author). 3 figs., 3 refs.

Subpicosecond oxygen trapping in the heme pocket of the oxygen sensor FixL observed by time-resolved resonance Raman spectroscopy.

Science.gov (United States)

Kruglik, Sergei G; Jasaitis, Audrius; Hola, Klara; Yamashita, Taku; Liebl, Ursula; Martin, Jean-Louis; Vos, Marten H

2007-05-01

Dissociation of oxygen from the heme domain of the bacterial oxygen sensor protein FixL constitutes the first step in hypoxia-induced signaling. In the present study, the photodissociation of the heme-O2 bond was used to synchronize this event, and time-resolved resonance Raman (TR(3)) spectroscopy with subpicosecond time resolution was implemented to characterize the heme configuration of the primary photoproduct. TR(3) measurements on heme-oxycomplexes are highly challenging and have not yet been reported. Whereas in all other known six-coordinated heme protein complexes with diatomic ligands, including the oxymyoglobin reported here, heme iron out-of-plane motion (doming) occurs faster than 1 ps after iron-ligand bond breaking; surprisingly, no sizeable doming is observed in the oxycomplex of the Bradyrhizobium japonicum FixL sensor domain (FixLH). This assessment is deduced from the absence of the iron-histidine band around 217 cm(-1) as early as 0.5 ps. We suggest that efficient ultrafast oxygen rebinding to the heme occurs on the femtosecond time scale, thus hindering heme doming. Comparing WT oxy-FixLH, mutant proteins FixLH-R220H and FixLH-R220Q, the respective carbonmonoxy-complexes, and oxymyoglobin, we show that a hydrogen bond of the terminal oxygen atom with the residue in position 220 is responsible for the observed behavior; in WT FixL this residue is arginine, crucially implicated in signal transmission. We propose that the rigid O2 configuration imposed by this residue, in combination with the hydrophobic and constrained properties of the distal cavity, keep dissociated oxygen in place. These results uncover the origin of the "oxygen cage" properties of this oxygen sensor protein.

Kinetics of Nif gene expression in a nitrogen-fixing bacterium.

Science.gov (United States)

Poza-Carrión, César; Jiménez-Vicente, Emilio; Navarro-Rodríguez, Mónica; Echavarri-Erasun, Carlos; Rubio, Luis M

2014-02-01

Nitrogen fixation is a tightly regulated trait. Switching from N2 fixation-repressing conditions to the N2-fixing state is carefully controlled in diazotrophic bacteria mainly because of the high energy demand that it imposes. By using quantitative real-time PCR and quantitative immunoblotting, we show here how nitrogen fixation (nif) gene expression develops in Azotobacter vinelandii upon derepression. Transient expression of the transcriptional activator-encoding gene, nifA, was followed by subsequent, longer-duration waves of expression of the nitrogenase biosynthetic and structural genes. Importantly, expression timing, expression levels, and NifA dependence varied greatly among the nif operons. Moreover, the exact concentrations of Nif proteins and their changes over time were determined for the first time. Nif protein concentrations were exquisitely balanced, with FeMo cofactor biosynthetic proteins accumulating at levels 50- to 100-fold lower than those of the structural proteins. Mutants lacking nitrogenase structural genes or impaired in FeMo cofactor biosynthesis showed overenhanced responses to derepression that were proportional to the degree of nitrogenase activity impairment, consistent with the existence of at least two negative-feedback regulatory mechanisms. The first such mechanism responded to the levels of fixed nitrogen, whereas the second mechanism appeared to respond to the levels of the mature NifDK component. Altogether, these findings provide a framework to engineer N2 fixation in nondiazotrophs.

Immobilization of ferrocene-modified SNAP-fusion proteins

NARCIS (Netherlands)

Wasserberg, D.; Uhlenheuer, D.; Neirynck, P.; Neirynck, Pauline; Cabanas Danés, Jordi; Schenkel, J.H.; Ravoo, B.J.; An, Q.; Huskens, Jurriaan; Milroy, L.G.; Brunsveld, Luc; Jonkheijm, Pascal

2013-01-01

The supramolecular assembly of proteins on surfaces has been investigated via the site-selective incorporation of a supramolecular moiety on proteins. To this end, fluorescent proteins have been site-selectively labeled with ferrocenes, as supramolecular guest moieties, via SNAP-tag technology. The

Products of random matrices from fixed trace and induced Ginibre ensembles

Science.gov (United States)

Akemann, Gernot; Cikovic, Milan

2018-05-01

We investigate the microcanonical version of the complex induced Ginibre ensemble, by introducing a fixed trace constraint for its second moment. Like for the canonical Ginibre ensemble, its complex eigenvalues can be interpreted as a two-dimensional Coulomb gas, which are now subject to a constraint and a modified, collective confining potential. Despite the lack of determinantal structure in this fixed trace ensemble, we compute all its density correlation functions at finite matrix size and compare to a fixed trace ensemble of normal matrices, representing a different Coulomb gas. Our main tool of investigation is the Laplace transform, that maps back the fixed trace to the induced Ginibre ensemble. Products of random matrices have been used to study the Lyapunov and stability exponents for chaotic dynamical systems, where the latter are based on the complex eigenvalues of the product matrix. Because little is known about the universality of the eigenvalue distribution of such product matrices, we then study the product of m induced Ginibre matrices with a fixed trace constraint—which are clearly non-Gaussian—and M  ‑  m such Ginibre matrices without constraint. Using an m-fold inverse Laplace transform, we obtain a concise result for the spectral density of such a mixed product matrix at finite matrix size, for arbitrary fixed m and M. Very recently local and global universality was proven by the authors and their coworker for a more general, single elliptic fixed trace ensemble in the bulk of the spectrum. Here, we argue that the spectral density of mixed products is in the same universality class as the product of M independent induced Ginibre ensembles.

(0,2) SCFTs from the Leigh-Strassler fixed point

Energy Technology Data Exchange (ETDEWEB)

Bobev, Nikolay [Perimeter Institute for Theoretical Physics, 31 Caroline Street North, Waterloo, ON N2L 2Y5 (Canada); Pilch, Krzysztof; Vasilakis, Orestis [Department of Physics and Astronomy, University of Southern California, Los Angeles, CA 90089 (United States)

2014-06-17

We show that there is a family of two-dimensional (0,2) SCFTs associated with twisted compactifications of the four-dimensional N=1 Leigh-Strassler fixed point on a closed hyperbolic Riemann surface. We calculate the central charges for this class of theories using anomalies and c-extremization. In a suitable truncation of the five-dimensional maximal supergravity, we construct supersymmetric AdS{sub 3} solutions that are holographic duals of those two-dimensional (0,2) SCFTs. We also exhibit supersymmetric domain wall solutions that are holographically dual to the RG flows between the four-dimensional and two-dimensional theories.

(0,2) SCFTs from the Leigh-Strassler fixed point

International Nuclear Information System (INIS)

Bobev, Nikolay; Pilch, Krzysztof; Vasilakis, Orestis

2014-01-01

We show that there is a family of two-dimensional (0,2) SCFTs associated with twisted compactifications of the four-dimensional N=1 Leigh-Strassler fixed point on a closed hyperbolic Riemann surface. We calculate the central charges for this class of theories using anomalies and c-extremization. In a suitable truncation of the five-dimensional maximal supergravity, we construct supersymmetric AdS 3 solutions that are holographic duals of those two-dimensional (0,2) SCFTs. We also exhibit supersymmetric domain wall solutions that are holographically dual to the RG flows between the four-dimensional and two-dimensional theories.

Adhesives for fixed orthodontic brackets.

Science.gov (United States)

Mandall, N A; Millett, D T; Mattick, C R; Hickman, J; Macfarlane, T V; Worthington, H V

2003-01-01

Bonding of orthodontic brackets to teeth is important to enable effective and efficient treatment with fixed appliances. The problem is bracket failure during treatment which increases operator chairside time and lengthens treatment time. A prolonged treatment is likely to increase the oral health risks of orthodontic treatment with fixed appliances one of which is irreversible enamel decalcification. To evaluate the effectiveness of different orthodontic adhesives for bonding. Electronic databases: the Cochrane Oral Health Group's Trials Register, the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE and EMBASE. Date of most recent searches: August 2002 (CENTRAL) (The Cochrane Library Issue 2, 2002). Trials were selected if they met the following criteria: randomised controlled trials (RCTs) and controlled clinical trials (CCTs) comparing two different adhesive groups. Participants were patients with fixed orthodontic appliances. The interventions were adhesives that bonded stainless steel brackets to all teeth except the molars. The primary outcome was debond or bracket failure. Data were recorded on decalcification as a secondary outcome, if present. Information regarding methods, participants, interventions, outcome measures and results were extracted in duplicate by pairs of reviewers (Nicky Mandall (NM) and Rye Mattick (CRM); Declan Millett (DTM) and Joy Hickman (JH2)). Since the data were not presented in a form that was amenable to meta-analysis, the results of the review are presented in narrative form only. Three trials satisfied the inclusion criteria. A chemical cured composite was compared with a light cure composite (one trial), a conventional glass ionomer cement (one trial) and a polyacid-modified resin composite (compomer) (one trial). The quality of the trial reports was generally poor. It is difficult to draw any conclusions from this review, however, suggestions are made for methods of improving future research involving

An OGA-Resistant Probe Allows Specific Visualization and Accurate Identification of O-GlcNAc-Modified Proteins in Cells.

Science.gov (United States)

Li, Jing; Wang, Jiajia; Wen, Liuqing; Zhu, He; Li, Shanshan; Huang, Kenneth; Jiang, Kuan; Li, Xu; Ma, Cheng; Qu, Jingyao; Parameswaran, Aishwarya; Song, Jing; Zhao, Wei; Wang, Peng George

2016-11-18

O-linked β-N-acetyl-glucosamine (O-GlcNAc) is an essential and ubiquitous post-translational modification present in nucleic and cytoplasmic proteins of multicellular eukaryotes. The metabolic chemical probes such as GlcNAc or GalNAc analogues bearing ketone or azide handles, in conjunction with bioorthogonal reactions, provide a powerful approach for detecting and identifying this modification. However, these chemical probes either enter multiple glycosylation pathways or have low labeling efficiency. Therefore, selective and potent probes are needed to assess this modification. We report here the development of a novel probe, 1,3,6-tri-O-acetyl-2-azidoacetamido-2,4-dideoxy-d-glucopyranose (Ac 3 4dGlcNAz), that can be processed by the GalNAc salvage pathway and transferred by O-GlcNAc transferase (OGT) to O-GlcNAc proteins. Due to the absence of a hydroxyl group at C4, this probe is less incorporated into α/β 4-GlcNAc or GalNAc containing glycoconjugates. Furthermore, the O-4dGlcNAz modification was resistant to the hydrolysis of O-GlcNAcase (OGA), which greatly enhanced the efficiency of incorporation for O-GlcNAcylation. Combined with a click reaction, Ac 3 4dGlcNAz allowed the selective visualization of O-GlcNAc in cells and accurate identification of O-GlcNAc-modified proteins with LC-MS/MS. This probe represents a more potent and selective tool in tracking, capturing, and identifying O-GlcNAc-modified proteins in cells and cell lysates.

Fixing the Sky: Why the History of Climate Engineering Matters (Invited)

Science.gov (United States)

Fleming, J. R.

2010-12-01

What shall we do about climate change? Is a planetary-scale technological fix possible or desirable? The joint AMS and AGU “Policy Statement on Geoengineering the Climate System” (2009) recommends “Coordinated study of historical, ethical, legal, and social implications of geoengineering that integrates international, interdisciplinary, and intergenerational issues and perspectives and includes lessons from past efforts to modify weather and climate.” I wrote Fixing the Sky: The Checkered History of Weather and Climate Control (Columbia University Press, 2010) with this recommendation in mind, to be fully accessible to scientists, policymakers, and the general public, while meeting or exceeding the scholarly standards of history. It is my intent, with this book, to bring history to bear on public policy issues.

The importance of a hot-sequential mechanism in triplet-state formation by charge recombination in reaction centers of bacterial photosynthesis

International Nuclear Information System (INIS)

Saito, K.; Mukai, K.; Sumi, H.

2006-01-01

In photosynthesis, pigment-excitation energies in the antenna system produced by light harvesting are transferred among antenna pigments toward the core antenna, where they are captured by the reaction center and initially fixed in the form of a charge separation. Primary charge separation between an oxidized special pair (P + ) and a reduced bacteriopheohytin (H - ) is occasionally intervened by recombination, and a spin-triplet state ( 3 P*) is formed on P in the bacterial reaction center. The 3 P* state is harmful to bio-organisms, inducing the formation of the highly damaging singlet oxygen species. Therefore, understanding the 3 P*-formation mechanism is important. The 3 P* formation is mediated by a state |m> of intermediate charge separation between P and the accessory chlorophyll, which is located between P and H. It will be shown theoretically in the present work that at room temperature, not only the mechanism of superexchange by quantum-mechanical virtual mediation at |m>, but also a hot-sequential mechanism contributes to the mediation. In the latter, although |m> is produced as a real state, the final state 3 P* is quickly formed during thermalization of phonons in the protein matrix in |m>. In the former, the final state is formed more quickly before dephasing-thermalization of phonons in |m>. 3 P* is unistep formed from the charge-separated state in the both mechanisms

Physical Activity Modifies the Association between Dietary Protein and Lean Mass of Postmenopausal Women.

Science.gov (United States)

Martinez, Jessica A; Wertheim, Betsy C; Thomson, Cynthia A; Bea, Jennifer W; Wallace, Robert; Allison, Matthew; Snetselaar, Linda; Chen, Zhao; Nassir, Rami; Thompson, Patricia A

2017-02-01

Maintenance of lean muscle mass and related strength is associated with lower risk for numerous chronic diseases of aging in women. Our aim was to evaluate whether the association between dietary protein and lean mass differs by physical activity level, amino acid composition, and body mass index categories. We performed a cross-sectional analysis of a prospective cohort. Participants were postmenopausal women from the Women's Health Initiative with body composition measurements by dual-energy x-ray absorptiometry (n=8,298). Our study measured percent lean mass, percent fat mass, and lean body mass index. Linear regression models adjusted for scanner serial number, age, calibrated energy intake, race/ethnicity, neighborhood socioeconomic status, and recreational physical activity were used to determine the relationship between protein intake and body composition measures. Likelihood ratio tests and stratified analysis were used to investigate physical activity and body mass index as potential effect modifiers. Biomarker-calibrated protein intake was positively associated with percent lean mass; women in the highest protein quintile had 6.3 percentage points higher lean mass than the lowest quintile (Plean body mass index were both inversely related to protein intake (both Plean body mass index (P interaction =0.011). Leucine intake was associated with lean mass, as were branched chain amino acids combined (both Plean mass in postmenopausal women. Importantly, those that also engage in physical activity have the highest lean mass across body mass index categories. Copyright © 2017 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

Analysis of some greedy algorithms for the single-sink fixed-charge transportation problem

DEFF Research Database (Denmark)

Görtz, Simon; Klose, Andreas

2009-01-01

-charge transportation problem. Nevertheless, just a few methods for solving this problem have been proposed in the literature. In this paper, some greedy heuristic solutions methods for the SSFCTP are investigated. It is shown that two greedy approaches for the SSFCTP known from the literature can be arbitrarily bad......, whereas an approximation algorithm proposed in the literature for the binary min-knapsack problem has a guaranteed worst case bound if adapted accordingly to the case of the SSFCTP....

An ontology-based search engine for protein-protein interactions.

Science.gov (United States)

Park, Byungkyu; Han, Kyungsook

2010-01-18

Keyword matching or ID matching is the most common searching method in a large database of protein-protein interactions. They are purely syntactic methods, and retrieve the records in the database that contain a keyword or ID specified in a query. Such syntactic search methods often retrieve too few search results or no results despite many potential matches present in the database. We have developed a new method for representing protein-protein interactions and the Gene Ontology (GO) using modified Gödel numbers. This representation is hidden from users but enables a search engine using the representation to efficiently search protein-protein interactions in a biologically meaningful way. Given a query protein with optional search conditions expressed in one or more GO terms, the search engine finds all the interaction partners of the query protein by unique prime factorization of the modified Gödel numbers representing the query protein and the search conditions. Representing the biological relations of proteins and their GO annotations by modified Gödel numbers makes a search engine efficiently find all protein-protein interactions by prime factorization of the numbers. Keyword matching or ID matching search methods often miss the interactions involving a protein that has no explicit annotations matching the search condition, but our search engine retrieves such interactions as well if they satisfy the search condition with a more specific term in the ontology.

Spatial cluster analysis of nanoscopically mapped serotonin receptors for classification of fixed brain tissue

Science.gov (United States)

Sams, Michael; Silye, Rene; Göhring, Janett; Muresan, Leila; Schilcher, Kurt; Jacak, Jaroslaw

2014-01-01

We present a cluster spatial analysis method using nanoscopic dSTORM images to determine changes in protein cluster distributions within brain tissue. Such methods are suitable to investigate human brain tissue and will help to achieve a deeper understanding of brain disease along with aiding drug development. Human brain tissue samples are usually treated postmortem via standard fixation protocols, which are established in clinical laboratories. Therefore, our localization microscopy-based method was adapted to characterize protein density and protein cluster localization in samples fixed using different protocols followed by common fluorescent immunohistochemistry techniques. The localization microscopy allows nanoscopic mapping of serotonin 5-HT1A receptor groups within a two-dimensional image of a brain tissue slice. These nanoscopically mapped proteins can be confined to clusters by applying the proposed statistical spatial analysis. Selected features of such clusters were subsequently used to characterize and classify the tissue. Samples were obtained from different types of patients, fixed with different preparation methods, and finally stored in a human tissue bank. To verify the proposed method, samples of a cryopreserved healthy brain have been compared with epitope-retrieved and paraffin-fixed tissues. Furthermore, samples of healthy brain tissues were compared with data obtained from patients suffering from mental illnesses (e.g., major depressive disorder). Our work demonstrates the applicability of localization microscopy and image analysis methods for comparison and classification of human brain tissues at a nanoscopic level. Furthermore, the presented workflow marks a unique technological advance in the characterization of protein distributions in brain tissue sections.

Protein nanoparticles for therapeutic protein delivery.

Science.gov (United States)

Herrera Estrada, L P; Champion, J A

2015-06-01

Therapeutic proteins can face substantial challenges to their activity, requiring protein modification or use of a delivery vehicle. Nanoparticles can significantly enhance delivery of encapsulated cargo, but traditional small molecule carriers have some limitations in their use for protein delivery. Nanoparticles made from protein have been proposed as alternative carriers and have benefits specific to therapeutic protein delivery. This review describes protein nanoparticles made by self-assembly, including protein cages, protein polymers, and charged or amphipathic peptides, and by desolvation. It presents particle fabrication and delivery characterization for a variety of therapeutic and model proteins, as well as comparison of the features of different protein nanoparticles.

Expression of PAT and NPT II proteins during the developmental stages of a genetically modified pepper developed in Korea.

Science.gov (United States)

Kim, Hyo Jin; Lee, Si Myung; Kim, Jae Kwang; Ryu, Tae Hun; Suh, Seok Cheol; Cho, Hyun Suk

2010-10-27

Estimation of the protein levels introduced in a biotechnology-derived product is conducted as part of an overall safety assessment. An enzyme-linked immunosorbent assay (ELISA) was used to analyze phosphinothricin acetyltransferase (PAT) and neomycin phosphotransferase II (NPT II) protein expression in a genetically modified (GM) pepper plant developed in Korea. PAT and NPT II expression levels, based on both dry weight and fresh weight, were variable among different plant generations and plant sections from isolated genetically modified organism (GMO) fields at four developmental stages. PAT expression was highest in leaves at anthesis (11.44 μg/gdw and 2.17 μg/gfw) and lowest in roots (0.12 μg/gdw and 0.01 μg/gfw). NPT II expression was also highest in leaves at anthesis (17.31 μg/gdw and 3.41 μg/gfw) and lowest in red pepper (0.65 μg/gdw and 0.12 μg/gfw). In pollen, PAT expression was 0.59-0.62 μg/gdw, while NPT II was not detected. Both PAT and NPT II showed a general pattern of decreased expression with progression of the growing season. As expected, PAT and NPT II protein expression was not detectable in control pepper plants.

Super spin-charge separation for class A, C and D disorder

International Nuclear Information System (INIS)

LeClair, Andre; Robinson, Dean J

2008-01-01

We prove versions of super spin-charge separation for all three of the symmetry groups SU(N), Sp(2N) and SO(N) of disordered Dirac fermions in 2 + 1 dimensions, which involve the supercurrent algebras gl(1|1) N ,osp(2|2) -2N and osp(2|2) N respectively. For certain restricted classes of disordered potentials, the latter supercurrent algebra based conformal field theories can arise as non-trivial low energy fixed points. For all cases with such a fixed point, we compute the density of states exponents as a function of N. (fast track communication)

On Risk Charges and Shadow Account Options in Pension Funds

DEFF Research Database (Denmark)

Jørgensen, Peter Løchte; Gatzert, Nadine

2015-01-01

to equityholders and our paper develops a model in which the influence of risk charges and shadow account options on stakeholders’ value can be quantified and studied. Our numerical results show that the value of shadow account options can be significant and thus come at the risk of expropriating policyholder......This paper studies the economic implications of regulatory systems which allow equityholders of pension companies to not only charge a specific premium to compensate them for their higher risk (compared to policyholders), but also to accumulate these risk charges in a so-called shadow account...... wealth. However, our analysis also shows that this risk can be remedied if proper attention is given to the specific contract design and to the fixing of fair contract parameters at the outset....

Texturized dairy proteins.

Science.gov (United States)

Onwulata, Charles I; Phillips, John G; Tunick, Michael H; Qi, Phoebi X; Cooke, Peter H

2010-03-01

Dairy proteins are amenable to structural modifications induced by high temperature, shear, and moisture; in particular, whey proteins can change conformation to new unfolded states. The change in protein state is a basis for creating new foods. The dairy products, nonfat dried milk (NDM), whey protein concentrate (WPC), and whey protein isolate (WPI) were modified using a twin-screw extruder at melt temperatures of 50, 75, and 100 degrees C, and moistures ranging from 20 to 70 wt%. Viscoelasticity and solubility measurements showed that extrusion temperature was a more significant (P extruded dairy protein ranged from rigid (2500 N) to soft (2.7 N). Extruding at or above 75 degrees C resulted in increased peak force for WPC (138 to 2500 N) and WPI (2.7 to 147.1 N). NDM was marginally texturized; the presence of lactose interfered with its texturization. WPI products extruded at 50 degrees C were not texturized; their solubility values ranged from 71.8% to 92.6%. A wide possibility exists for creating new foods with texturized dairy proteins due to the extensive range of states achievable. Dairy proteins can be used to boost the protein content in puffed snacks made from corn meal, but unmodified, they bind water and form doughy pastes with starch. To minimize the water binding property of dairy proteins, WPI, or WPC, or NDM were modified by extrusion processing. Extrusion temperature conditions were adjusted to 50, 75, or 100 degrees C, sufficient to change the structure of the dairy proteins, but not destroy them. Extrusion modified the structures of these dairy proteins for ease of use in starchy foods to boost nutrient levels. Dairy proteins can be used to boost the protein content in puffed snacks made from corn meal, but unmodified, they bind water and form doughy pastes with starch. To minimize the water binding property of dairy proteins, whey protein isolate, whey protein concentrate, or nonfat dried milk were modified by extrusion processing. Extrusion

Amperometric Immunosensor Based on a Protein A/Deposited Gold Nanocrystals Modified Electrode for Carbofuran Detection

Directory of Open Access Journals (Sweden)

Xia Sun

2011-12-01

Full Text Available In this paper, an amperometric immunosensor modified with protein A/deposited gold nanocrystals (DpAu was developed for the ultrasensitive detection of carbofuran residues. First, DpAu were electrodeposited onto the Au electrode surface to absorb protein A (PA and improve the electrode conductivity. Then PA was dropped onto the surface of DpAu film, used for binding antibody Fc fragments. Next, anti-carbofuran monoclonal antibody was immobilized on the PA modified electrode. Finally, bovine serum albumin (BSA was employed to block the possible remaining active sites avoiding any nonspecific adsorption. The fabrication procedure of the immunosensor was characterized by electrochemical impedance spectroscopy (EIS and cyclic voltammetry (CV, respectively. With the excellent electroconductivity of DpAu and the PA’s oriented immobilization of antibodies, a highly efficient immuno-reaction and detection sensitivity could be achieved. The influences of the electrodeposition time of DpAu, pH of the detection solution and incubation time on the current response of the fabricated immunosensor were investigated. Under optimized conditions, the current response was proportional to the concentration of carbofuran which ranged from 1 to 100 ng/mL and 100 ng/mL to 100 μg/mL. The detection limit was 0.1924 ng/mL. The proposed carbofuran immnuosensor exhibited high specificity, reproducibility, stability and regeneration performance, which may open a new door for ultrasensitive detection of carbofuran residues in vegetables and fruits.

Senp1 Is Essential for Desumoylating Sumo1-Modified Proteins but Dispensable for Sumo2 and Sumo3 Deconjugation in the Mouse Embryo

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Prashant Sharma

2013-05-01

Full Text Available Posttranslational modification with small ubiquitin-like modifier (Sumo regulates numerous cellular and developmental processes. Sumoylation is dynamic with deconjugation by Sumo-specific proteases (Senps regulating steady-state levels. Different Senps are found in distinct subcellular domains, which may limit their deconjugation activity to colocalizing Sumo-modified proteins. In vitro, Senps can discriminate between the different Sumo paralogs: Sumo1 versus the highly related Sumo2 and Sumo3 (Sumo2/3, which can form poly-Sumo chains. However, a full understanding of Senp specificity in vivo is still lacking. Here, using biochemical and genetic approaches, we establish that Senp1 has an essential, nonredundant function to desumoylate Sumo1-modified proteins during mouse embryonic development. Senp1 specificity for Sumo1 conjugates represents an intrinsic function and not simply a product of colocalization. In contrast, Senp1 has only a limited role in Sumo2/3 desumoylation, although it may regulate Sumo1-mediated termination of poly-Sumo2/3 chains.

Moment-Preserving Computational Approach for High Energy Charged Particle Transport

Science.gov (United States)

2016-05-16

posed, but with modified cross sections such that the resulting single-event Monte Carlo simulation is computationally efficient (minutes vs . days...configurations, which are all characteristics of real world applications. In other words , it is possible to simulate real, physical phenomena using charged...0 < 0.95) ~ 1 2() ≫ 1, (3) Demonstrating that scattering is highly forward peaked. Thus, the picture of charged particle interactions

Utilization of nitrogen fixing trees

Energy Technology Data Exchange (ETDEWEB)

Brewbaker, J.L.; Beldt, R. van den; MacDicken, K.; Budowski, G.; Kass, D.C.L.; Russo, R.O.; Escalante, G.; Herrera, R.; Aranguren, J.; Arkcoll, D.B.; Doebereinger, J. (cord.)

1983-01-01

Six papers from the symposium are noted. Brewbaker, J.L., Beldt, R. van den, MacDicken, K. Fuelwood uses and properties of nitrogen-fixing trees, pp 193-204, (Refs. 15). Includes a list of 35 nitrogen-fixing trees of high fuelwood value. Budowski, G.; Kass, D.C.L.; Russo, R.O. Leguminous trees for shade, pp 205-222, (Refs. 68). Escalante, G., Herrera, R., Aranguren, J.; Nitrogen fixation in shade trees (Erythrina poeppigiana) in cocoa plantations in northern Venezuela, pp 223-230, (Refs. 13). Arkcoll, D.B.; Some leguminous trees providing useful fruits in the North of Brazil, pp 235-239, (Refs. 13). This paper deals with Parkia platycephala, Pentaclethra macroloba, Swartzia sp., Cassia leiandra, Hymenaea courbaril, dipteryz odorata, Inga edulis, I. macrophylla, and I. cinnamonea. Baggio, A.J.; Possibilities of the use of Gliricidia sepium in agroforestry systems in Brazil, pp 241-243; (Refs. 15). Seiffert, N.F.; Biological nitrogen and protein production of Leucaena cultivars grown to supplement the nutrition of ruminants, pp 245-249, (Refs. 14). Leucaena leucocephala cv. Peru, L. campina grande (L. leucocephala), and L. cunningham (L. leucocephalae) were promising for use as browse by beef cattle in central Brazil.

Factors that influence the design of modified clays - or how knowing your clay can save your day

International Nuclear Information System (INIS)

Gates, W.P.; Slade, P.G.

1998-01-01

Full text: Smectites vary greatly in their permanent layer charge characteristics, including total charge, distribution of charge between tetrahedral and octahedral sheets and heterogeneity of charge from flake to flake. Smectites and vermiculites are different from the micaceous layer silicates in their ability to swell by the uptake of cations and polar and non polar solvents. Vermiculites differ from the smectites predominantly in the large contribution of tetrahedrally located charge relative to their total layer charge density. Understanding of the complex relations between layer charge and interlayer space of clay mineral surfaces can be applied toward the design of optimal organically modified clays suitable for environmental and industrial uses. In general, it is known that smectite charge density dictates the total amount of modifying organic cation that can be added to a particular clay, the orientation that the organic cation adopts within the interlayer spaces with respect to the siloxane surfaces of the clay and ultimately, the capacity of specific, organically modified clay to imbibe contaminants or other compounds. These same properties are dependent on the size and configuration of the modifying organic cation(s) as well as the percentage of the exchange capacity utilised, and thus, the amount of specific surface of the clay that is covered by the modifying organic cation. All these factors must be kept in mind in the design of inexpensive and useful modified clays. This paper reports on the application of polarised FT-IR and X-ray diffraction methods to the observation that layer charge density governs the orientation of trimethylphenylammonium (TMPA) cations in the interlayer space of smectites and vermiculites. The TMPA exchanged forms of several smectites and vermiculites were studied, whose layer charges ranged between X=0.37 and X=0.95 e - per formula unit and in which the location of charge varied with respect to the octahedral and

Deactivation of nickel hydroxide-gold modified electrodes

OpenAIRE

Caram, Bruno; Tucceri, Ricardo

2013-01-01

The aim of the present work was to study how the charge-transport process of a nickel hydroxide film electrochemically synthesized on a gold substrate is modified when the electrode is stored for a long time. It was found that nickel hydroxide films are deactivated under storage, that is, films became less conductive than films immediately prepared (nondeactivated). This study was carried out in the context of the rotating disc electrode voltammetry when the modified electrode contacts an ele...

Physicochemical and Antioxidant Properties of Buckwheat Protein Isolates with Different Polyphenolic Content Modified by Limited Hydrolysis with Trypsin

Directory of Open Access Journals (Sweden)

Xiao-Yan Wang

2012-01-01

Full Text Available Effects of limited hydrolysis with trypsin on the physicochemical and antioxidant properties of buckwheat protein isolates (BPIs obtained with untreated and 2-propanol-extracted meal have been investigated and compared. The dephenolization treatment significantly improved the hydrolysis of BPI, which resulted in the gradual decrease in total and protein-bound polyphenolic content, but an increase in the free polyphenolic content. The hydrolysis of globulins was much easier than that of the albumins. The removal of polyphenols improved the hydrolysis of the albumin fraction. The modified BPIs with high polyphenolic content exhibited much higher DPPH radical scavenging activity and reducing power, but poorer ferrous ion chelating ability than those with low polyphenolic content. These results suggest that the limited hydrolysis is suitable for modification of the properties of buckwheat proteins.

Charge collection and charge pulse formation in highly irradiated silicon planar detectors

International Nuclear Information System (INIS)

Dezillie, B.; Li, Z.; Eremin, V.

1998-06-01

The interpretation of experimental data and predictions for future experiments for high-energy physics have been based on conventional methods like capacitance versus voltage (C-V) measurements. Experiments carried out on highly irradiated detectors show that the kinetics of the charge collection and the dependence of the charge pulse amplitude on the applied bias are deviated too far from those predicted by the conventional methods. The described results show that in highly irradiated detectors, at a bias lower than the real full depletion voltage (V fd ), the kinetics of the charge collection (Q) contains a fast and a slow component. At V = V fd *, which is the full depletion voltage traditionally determined by the extrapolation of the fast component amplitude of q versus bias to the maximum value or from the standard C-V measurements, the pulse has a slow component with significant amplitude. This slow component can only be eliminated by applying additional bias that amounts to the real full depletion voltage (V fd ) or more. The above mentioned regularities are explained in this paper in terms of a model of an irradiated detector with multiple regions. This model allows one to use C-V, in a modified way, as well as TChT (transient charge technique) measurements to determine the V fd for highly irradiated detectors

Mitochondrial associated ubiquitin fold modifier-1 mediated protein conjugation in Leishmania donovani.

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Sreenivas Gannavaram

2011-01-01

Full Text Available In this report, we demonstrate the existence of the ubiquitin fold modifier-1 (Ufm1 and its conjugation pathway in trypanosomatid parasite Leishmania donovani. LdUfm1 is activated by E1-like enzyme LdUba5. LdUfc1 (E2 specifically interacted with LdUfm1 and LdUba5 to conjugate LdUfm1 to proteinaceous targets. Mass spectrometry analysis revealed that LdUfm1 is conjugated to Leishmania protein targets that are associated with mitochondria. Immunofluorescence experiments showed that Leishmania Ufm1, Uba5 and Ufc1 are associated with the mitochondria. The demonstration that all the components of this system as well as the substrates are associated with mitochondrion suggests it may have physiological roles not yet described in any other organism. Overexpression of a non-conjugatable form of LdUfm1 and an active site mutant of LdUba5 resulted in reduced survival of Leishmania in the macrophage. Since mitochondrial activities are developmentally regulated in the life cycle of trypanosomatids, Ufm1 mediated modifications of mitochondrial proteins may be important in such regulation. Thus, Ufm1 conjugation pathway in Leishmania could be explored as a potential drug target in the control of Leishmaniasis.

Complex fluids with mobile charge-regulating macro-ions

Science.gov (United States)

Markovich, Tomer; Andelman, David; Podgornik, Rudi

2017-10-01

We generalize the concept of charge regulation of ionic solutions, and apply it to complex fluids with mobile macro-ions having internal non-electrostatic degrees of freedom. The suggested framework provides a convenient tool for investigating systems where mobile macro-ions can self-regulate their charge (e.g., proteins). We show that even within a simplified charge-regulation model, the charge dissociation equilibrium results in different and notable properties. Consequences of the charge regulation include a positional dependence of the effective charge of the macro-ions, a non-monotonic dependence of the effective Debye screening length on the concentration of the monovalent salt, a modification of the electric double-layer structure, and buffering by the macro-ions of the background electrolyte.

Selective determination of dopamine using quantum-sized gold nanoparticles protected with charge selective ligands

Science.gov (United States)

Kwak, Kyuju; Kumar, S. Senthil; Lee, Dongil

2012-06-01

We report here the selective determination of dopamine (DA) using quantum-sized gold nanoparticles coated with charge selective ligands. Glutathione protected gold nanoparticles (GS-Au25) were synthesized and immobilized into a sol-gel matrix via thiol linkers. The GS-Au25 modified sol-gel electrode was found to show excellent electrocatalytic activity towards the oxidation of DA but no activity towards the oxidation of ascorbic acid. The role of electrostatic charge in the selective electrocatalytic activity of GS-Au25 was verified by voltammetry of redox markers carrying opposite charges. The pH dependent sensitivity for the determination of DA further confirmed the charge screening effect of GS-Au25. Mechanistic investigation revealed that the selectivity is attained by the selective formation of an electrostatic complex between the negatively charged GS-Au25 and DA cation. The GS-Au25 modified sol-gel electrode also showed excellent selectivity for DA in the presence of an interferent, ascorbic acid.We report here the selective determination of dopamine (DA) using quantum-sized gold nanoparticles coated with charge selective ligands. Glutathione protected gold nanoparticles (GS-Au25) were synthesized and immobilized into a sol-gel matrix via thiol linkers. The GS-Au25 modified sol-gel electrode was found to show excellent electrocatalytic activity towards the oxidation of DA but no activity towards the oxidation of ascorbic acid. The role of electrostatic charge in the selective electrocatalytic activity of GS-Au25 was verified by voltammetry of redox markers carrying opposite charges. The pH dependent sensitivity for the determination of DA further confirmed the charge screening effect of GS-Au25. Mechanistic investigation revealed that the selectivity is attained by the selective formation of an electrostatic complex between the negatively charged GS-Au25 and DA cation. The GS-Au25 modified sol-gel electrode also showed excellent selectivity for DA in the

A genetically modified protein-based hydrogel for 3D culture of AD293 cells.

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Xiao Du

Full Text Available Hydrogels have strong application prospects for drug delivery, tissue engineering and cell therapy because of their excellent biocompatibility and abundant availability as scaffolds for drugs and cells. In this study, we created hybrid hydrogels based on a genetically modified tax interactive protein-1 (TIP1 by introducing two or four cysteine residues in the primary structure of TIP1. The introduced cysteine residues were crosslinked with a four-armed poly (ethylene glycol having their arm ends capped with maleimide residues (4-armed-PEG-Mal to form hydrogels. In one form of the genetically modification, we incorporated a peptide sequence 'GRGDSP' to introduce bioactivity to the protein, and the resultant hydrogel could provide an excellent environment for a three dimensional cell culture of AD293 cells. The AD293 cells continued to divide and displayed a polyhedron or spindle-shape during the 3-day culture period. Besides, AD293 cells could be easily separated from the cell-gel constructs for future large-scale culture after being cultured for 3 days and treating hydrogel with trypsinase. This work significantly expands the toolbox of recombinant proteins for hydrogel formation, and we believe that our hydrogel will be of considerable interest to those working in cell therapy and controlled drug delivery.

Bremsstrahlung from Relativistic Heavy Ions in a Fixed Target Experiment at the LHC

International Nuclear Information System (INIS)

Mikkelsen, Rune E.; Uggerhøj, Ulrik I.; Sørensen, Allan H.

2015-01-01

We calculate the emission of bremsstrahlung from lead and argon ions in ultraperipheral collisions in a fixed target experiment (AFTER) that uses the LHC beams. With nuclear charges of Ze equal to 82e and 18e, respectively, these ions are accelerated to energies of 7 Tev × Z. The bremsstrahlung peaks around ≈100 GeV and the spectrum exposes the nuclear structure of the incoming ion. The peak structure is significantly different from the flat power spectrum pertaining to a point charge. Photons are predominantly emitted within an angle of 1/γ to the direction of ion propagation. Our calculations are based on the Weizsäcker-Williams method of virtual quanta with application of existing experimental data on photonuclear interactions.

Effect of reference conditions on flow rate, modifier fraction and retention in supercritical fluid chromatography.

Science.gov (United States)

De Pauw, Ruben; Shoykhet Choikhet, Konstantin; Desmet, Gert; Broeckhoven, Ken

2016-08-12

When using compressible mobile phases such as fluidic CO2, the density, the volumetric flow rates and volumetric fractions are pressure dependent. The pressure and temperature definition of these volumetric parameters (referred to as the reference conditions) may alter between systems, manufacturers and operating conditions. A supercritical fluid chromatography system was modified to operate in two modes with different definition of the eluent delivery parameters, referred to as fixed and variable mode. For the variable mode, the volumetric parameters are defined with reference to the pump operating pressure and actual pump head temperature. These conditions may vary when, e.g. changing the column length, permeability, flow rate, etc. and are thus variable reference conditions. For the fixed mode, the reference conditions were set at 150bar and 30°C, resulting in a mass flow rate and mass fraction of modifier definition which is independent of the operation conditions. For the variable mode, the mass flow rate of carbon dioxide increases with system pump operating pressure, decreasing the fraction of modifier. Comparing the void times and retention factor shows that the deviation between the two modes is almost independent of modifier percentage, but depends on the operating pressure. Recalculating the set volumetric fraction of modifier to the mass fraction results in the same retention behaviour for both modes. This shows that retention in SFC can be best modelled using the mass fraction of modifier. The fixed mode also simplifies method scaling as it only requires matching average column pressure. Copyright © 2016 Elsevier B.V. All rights reserved.

Emittance and damping of electrons in the neighborhood of resonance fixed points

International Nuclear Information System (INIS)

Crosbie, E.A.

1993-01-01

The stable fixed points generated by nonlinear field harmonics in a cyclic lattice define a multiturn stable orbit. The position of the orbit for each turn in each magnet of the lattice determines the betatron tunes and lattice dispersion functions describing the linear motion of charged particles with respect to the stable orbit. Since the position of the fixed points is dependent in part on the central orbit tune, it turns out that the multiturn orbit dispersion function depends to a large extent on the central orbit chromaticity. In particular, the horizontal partition number can be made to vary from values less than zero (horizontal antidamping for electrons) to values greater than three (longitudinal antidamping). The central orbit chromaticity therefore plays a major role in determining the characteristic emittance of an electron beam with respect to the multiturn orbit

A quantitative analysis of municipal solid waste disposal charges in China.

Science.gov (United States)

Wu, Jian; Zhang, Weiqian; Xu, Jiaxuan; Che, Yue

2015-03-01

Rapid industrialization and economic development have caused a tremendous increase in municipal solid waste (MSW) generation in China. China began implementing a policy of MSW disposal fees for household waste management at the end of last century. Three charging methods were implemented throughout the country: a fixed disposal fee, a potable water-based disposal fee, and a plastic bag-based disposal fee. To date, there has been little qualitative or quantitative analysis on the effectiveness of this relatively new policy. This paper provides a general overview of MSW fee policy in China, attempts to verify whether the policy is successful in reducing general waste collected, and proposes an improved charging system to address current problems. The paper presents an empirical statistical analysis of policy effectiveness derived from an environmental Kuznets curve (EKC) test on panel data of China. EKC tests on different kinds of MSW charge systems were then examined for individual provinces or cities. A comparison of existing charging systems was conducted using environmental and economic criteria. The results indicate the following: (1) the MSW policies implemented over the study period were effective in the reduction of waste generation, (2) the household waste discharge fee policy did not act as a strong driver in terms of waste prevention and reduction, and (3) the plastic bag-based disposal fee appeared to be performing well according to qualitative and quantitative analysis. Based on current situation of waste discharging management in China, a three-stage transitional charging scheme is proposed and both advantages and drawbacks discussed. Evidence suggests that a transition from a fixed disposal fee to a plastic bag-based disposal fee involving various stakeholders should be the next objective of waste reduction efforts.

Fixed Points

Indian Academy of Sciences (India)

Home; Journals; Resonance – Journal of Science Education; Volume 5; Issue 5. Fixed Points - From Russia with Love - A Primer of Fixed Point Theory. A K Vijaykumar. Book Review Volume 5 Issue 5 May 2000 pp 101-102. Fulltext. Click here to view fulltext PDF. Permanent link:

Immunotoxicological Evaluation of Genetically Modified Rice Expressing Cry1Ab/Ac Protein (TT51-1) by a 6-Month Feeding Study on Cynomolgus Monkeys

OpenAIRE

Tan, Xiaoyan; Zhou, Xiaobing; Tang, Yao; Lv, Jianjun; Zhang, Lin; Sun, Li; Yang, Yanwei; Miao, Yufa; Jiang, Hua; Chen, Gaofeng; Huang, Zhiying; Wang, Xue

2016-01-01

The present study was performed to evaluate the food safety of TT51-1, a new type of genetically modified rice that expresses the Cry1Ab/Ac protein (Bt toxin) and is highly resistant to most lepidopteran pests. Sixteen male and 16 female cynomolgus monkeys were randomly divided into four groups: conventional rice (non-genetically modified rice, non-GM rice), positive control, 17.5% genetically modified rice (GM rice) and 70% GM rice. Monkeys in the non-GM rice, positive control, and GM rice g...

Surface charge accumulation of particles containing radionuclides in open air.

Science.gov (United States)

Kim, Yong-Ha; Yiacoumi, Sotira; Tsouris, Costas

2015-05-01

Radioactivity can induce charge accumulation on radioactive particles. However, electrostatic interactions caused by radioactivity are typically neglected in transport modeling of radioactive plumes because it is assumed that ionizing radiation leads to charge neutralization. The assumption that electrostatic interactions caused by radioactivity are negligible is evaluated here by examining charge accumulation and neutralization on particles containing radionuclides in open air. A charge-balance model is employed to predict charge accumulation on radioactive particles. It is shown that particles containing short-lived radionuclides can be charged with multiple elementary charges through radioactive decay. The presence of radioactive particles can significantly modify the particle charge distribution in open air and yield an asymmetric bimodal charge distribution, suggesting that strong electrostatic particle interactions may occur during short- and long-range transport of radioactive particles. Possible effects of transported radioactive particles on electrical properties of the local atmosphere are reported. The study offers insight into transport characteristics of airborne radionuclides. Results are useful in atmospheric transport modeling of radioactive plumes. Copyright © 2015 Elsevier Ltd. All rights reserved.

Fixing All Moduli in a Simple F-Theory Compactification

International Nuclear Information System (INIS)

Denef, F.

2005-01-01

We discuss a simple example of an F-theory compactification on a Calabi-Yau fourfold where background fluxes, together with nonperturbative effects from Euclidean D3 instantons and gauge dynamics on D7 branes, allow us to fix all closed and open string moduli. We explicitly check that the known higher order corrections to the potential, which we neglect in our leading approximation, only shift the results by a small amount. In our exploration of the model, we encounter interesting new phenomena, including examples of transitions where D7 branes absorb O3 planes, while changing topology to preserve the net D3 charge

In situ zymography and immunolabeling in fixed and decalcified craniofacial tissues.

Science.gov (United States)

Porto, Isabel M; Rocha, Lenaldo B; Rossi, Marcos A; Gerlach, Raquel F

2009-07-01

In situ zymography is a very important technique that shows the proteolytic activity in sections and allows researchers to observe the specific sites of proteolysis in tissues or cells. It is normally performed in non-fixed frozen sections and is not routinely performed in calcified tissues. In this study, we describe a technique that maintains proteolytic activity in fixed and decalcified sections obtained after routine paraffin sectioning in conventional microtome and cryostat sections. We used adult rat hemimandibles, which presented bone, enamel, and dentine matrices; the substrate used was dye-quenched-gelatin. Gelatinolytic activity was colocalized with MMP-2 using fluorescent antibodies. Specific proteolytic activity was observed in all sections, compatible with metalloproteinase activity, particularly in dentine and bone. Furthermore, matrix metalloproteinase-2 was colocalized to the sites of green fluorescence in dentine. In conclusion, the technique presented here will allow in situ zymography reactions in fixed, decalcified, and paraffin-embedded tissues, and we showed that paraformaldehyde-lysine-periodate-fixed cryostat sections are suitable for colocalization of gelatinolytic activity and protein labeling with antibodies.

Like-charge attraction in a one-dimensional setting: the importance of being odd

Science.gov (United States)

Trizac, Emmanuel; Téllez, Gabriel

2018-03-01

From cement cohesion to DNA condensation, a proper statistical physics treatment of systems with long-range forces is important for a number of applications in physics, chemistry, and biology. We compute here the effective force between fixed charged macromolecules, screened by oppositely charged mobile ions (counterions). We treat the problem in a one-dimensional configuration that allows for interesting discussion and derivation of exact results, remaining at a level of mathematical difficulty compatible with an undergraduate course. Emphasis is put on the counterintuitive but fundamental phenomenon of like-charge attraction, which our treatment brings for the first time to the level of undergraduate teaching. The parity of the number of counterions is shown to play a prominent role, which sheds light on the binding mechanism at work when like-charge macromolecules do attract.

A modified strategy for sequence specific assignment of protein NMR spectra based on amino acid type selective experiments

International Nuclear Information System (INIS)

Schubert, Mario; Labudde, Dirk; Leitner, Dietmar; Oschkinat, Hartmut; Schmieder, Peter

2005-01-01

The determination of the three-dimensional structure of a protein or the study of protein-ligand interactions requires the assignment of all relevant nuclei as an initial step. This is nowadays almost exclusively performed using triple-resonance experiments. The conventional strategy utilizes one or more pairs of three dimensional spectra to obtain redundant information and thus reliable assignments. Here, a modified strategy for obtaining sequence specific assignments based on two dimensional amino acid type selective triple-resonance experiments is proposed. These experiments can be recorded with good resolution in a relatively short time. They provide very specific and redundant information, in particular on sequential connectivities, that drastically increases the ease and reliability of the assignment procedure, done either manually or in an automated fashion. The new strategy is demonstrated with the protein domain PB1 from yeast CDC24p

Manipulation of stored charge in anodic aluminium oxide/SiO2 dielectric stacks by the use of pulsed anodisation

International Nuclear Information System (INIS)

Lu, Zhong; Ouyang, Zi; Grant, Nicholas; Wan, Yimao; Yan, Di; Lennon, Alison

2016-01-01

Graphical abstract: - Highlights: • Pulse anodisation was used to grow AAO layers with controllable stored charge. • Stored charge density ranging from −5.2 × 10 11 to 2.5 × 10 12 q/cm 2 was demonstrated. • Enhancement in surface passivation was demonstrated with charge management. • Annealing significantly reduces the positive stored charge and the interface defect. - Abstract: A method of fabricating anodic aluminium oxide (AAO) with the capability of manipulating its stored charge is reported. This method involves the use of a pulsed current source to anodise aluminium layers instead of the typically used constant current/voltage source, with the test structures experiencing positive and negative cycles periodically. By tuning the positive cycle percentage, it is demonstrated that the effective stored charge density can be manipulated in a range from −5.2 × 10 11 to 2.5 × 10 12 q/cm 2 when the AAO is formed over a 12 nm SiO 2 layer. An investigation of the stored charge distribution in the dielectric stacks indicates a positive fixed charge at the SiO 2 /Si interface, a negative fixed charge at the AAO/SiO 2 interface and a positive bulk charge within the AAO layer. The effective stored charge density and interface states were found to be affected by annealing conditions and it is suggested that oxygen annealing can reduce the bulk positive charge while post-metallisation anneal is most effective in reducing silicon interface defects. Charge manipulation using pulsed anodisation is shown to reduce carrier recombination on boron-diffused silicon surfaces highlighting the potential of the process to be used to tune the electrical properties of dielectric layers so that they can reduce surface recombination on silicon surfaces having different dopant polarity and concentrations.

Fourier Imaging X-ray Spectrometer (FIXS) for the Argentinian, Scout-launched satelite de Aplicaciones Cienficas-1 (SAC-1)

International Nuclear Information System (INIS)

Dennis, B.R.; Crannell, C.J.; Desai, U.D.

1988-01-01

The Fourier Imaging X-ray Spectrometer (FIXS) is one of four instruments on SAC-1, the Argentinian satellite being proposed for launch by NASA on a Scout rocket in 1992/3. The FIXS is designed to provide solar flare images at X-ray energies between 5 and 35 keV. Observations will be made on arcsecond size scales and subsecond time scales of the processes that modify the electron spectrum and the thermal distribution in flaring magnetic structures