Alanine/epr pellet dosimeter using poly(vinyl butyral-co-vinyl alcohol-co-vinyl acetate) copolymer as a binder for radiation dosimetry

International Nuclear Information System (INIS)

Beshir, W.B.; Ezz El-Din, H.M.; Abdel-fatth, A.A.; Ebraheem, S.

2005-01-01

A new alanine pellet dosimeter was developed for gamma and electron beam radiation dosimetry. Alanine powder was mixed with a new binding material, poly(vinyl butyral-co-vinyl alcohol-co-vinyl acetate) copolymer. Pellets were prepared by pressing fine powder alanine with 60% copolymer binder by using hydraulic press and a specially designed pressing die. The radiation-formed stable free radicals were analysed by using electron paramagnetic resonance (EPR) spectroscopy. The useful dose range of these pellets was found to ranges from 1 to 80 kGy. The stability of the radiation- induced response was also studied

Alpha-helical hydrophobic polypeptides form proton-selective channels in lipid bilayers

Science.gov (United States)

Oliver, A. E.; Deamer, D. W.

1994-01-01

Proton translocation is important in membrane-mediated processes such as ATP-dependent proton pumps, ATP synthesis, bacteriorhodopsin, and cytochrome oxidase function. The fundamental mechanism, however, is poorly understood. To test the theoretical possibility that bundles of hydrophobic alpha-helices could provide a low energy pathway for ion translocation through the lipid bilayer, polyamino acids were incorporated into extruded liposomes and planar lipid membranes, and proton translocation was measured. Liposomes with incorporated long-chain poly-L-alanine or poly-L-leucine were found to have proton permeability coefficients 5 to 7 times greater than control liposomes, whereas short-chain polyamino acids had relatively little effect. Potassium permeability was not increased markedly by any of the polyamino acids tested. Analytical thin layer chromatography measurements of lipid content and a fluorescamine assay for amino acids showed that there were approximately 135 polyleucine or 65 polyalanine molecules associated with each liposome. Fourier transform infrared spectroscopy indicated that a major fraction of the long-chain hydrophobic peptides existed in an alpha-helical conformation. Single-channel recording in both 0.1 N HCl and 0.1 M KCl was also used to determine whether proton-conducting channels formed in planar lipid membranes (phosphatidylcholine/phosphatidylethanolamine, 1:1). Poly-L-leucine and poly-L-alanine in HCl caused a 10- to 30-fold increase in frequency of conductive events compared to that seen in KCl or by the other polyamino acids in either solution. This finding correlates well with the liposome observations in which these two polyamino acids caused the largest increase in membrane proton permeability but had little effect on potassium permeability. Poly-L-leucine was considerably more conductive than poly-L-alanine due primarily to larger event amplitudes and, to a lesser extent, a higher event frequency. Poly-L-leucine caused two

Degradable polyphosphazene/poly(alpha-hydroxyester) blends: degradation studies.

Science.gov (United States)

Ambrosio, Archel M A; Allcock, Harry R; Katti, Dhirendra S; Laurencin, Cato T

2002-04-01

Biomaterials based on the polymers of lactic acid and glycolic acid and their copolymers are used or studied extensively as implantable devices for drug delivery, tissue engineering and other biomedical applications. Although these polymers have shown good biocompatibility, concerns have been raised regarding their acidic degradation products, which have important implications for long-term implantable systems. Therefore, we have designed a novel biodegradable polyphosphazene/poly(alpha-hydroxyester) blend whose degradation products are less acidic than those of the poly(alpha-hydroxyester) alone. In this study, the degradation characteristics of a blend of poly(lactide-co-glycolide) (50:50 PLAGA) and poly[(50% ethyl glycinato)(50% p-methylphenoxy) phosphazene] (PPHOS-EG50) were qualitatively and quantitatively determined with comparisons made to the parent polymers. Circular matrices (14mm diameter) of the PLAGA, PPHOS-EG50 and PLAGA-PPHOS-EG50 blend were degraded in non-buffered solutions (pH 7.4). The degraded polymers were characterized for percentage mass loss and molecular weight and the degradation medium was characterized for acid released in non-buffered solutions. The amounts of neutralizing base necessary to bring about neutral pH were measured for each polymer or polymer blend during degradation. The poly(phosphazene)/poly(lactide-co-glycolide) blend required significantly less neutralizing base in order to bring about neutral solution pH during the degradation period studied. The results indicated that the blend degraded at a rate intermediate to that of the parent polymers and that the degradation products of the polyphosphazene neutralized the acidic degradation products of PLAGA. Thus, results from these in vitro degradation studies suggest that the PLAGA-PPHOS-EG50 blend may provide a viable improvement to biomaterials based on acid-releasing organic polymers.

BAROMETRIC PRESSURE and Other Data from ALPHA HELIX From Chukchi Sea and Others from 19930709 to 19930807 (NODC Accession 9400062)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — The Conductivity, Temperature and Depth (CTD) and other data were collected in Bering Sea, Chukchi Sea. Data was collected from Ship ALPHA HELIX cruise Aleutian...

Temperature, salinity and transmissivity data from the Alpha Helix in the Arctic Ocean, 2000 - 2004 (NODC Accession 0059005)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — This is calibrated CTD downcast data from five Alpha Helix cruises: HX235, 8/1/2000 - 9/30/2000 HX250, 9/1/2001 - 9/30/2001 HX260, 6/1/2002 - 6/30/2002 HX274,...

Temperature, salinity, conductivity, pressure, transmissivity measurements collected using CTD from the Alpha Helix in the Chukchi Sea during 1996 (NODC Accession 0061042)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — Temperature, salinity, conductivity, pressure, and transmissivity data gathered by CTD from the Alpha Helix (cruise HX194), September 1996

BAROMETRIC PRESSURE and Other Data from ALPHA HELIX From North Pacific Ocean and Others from 19931016 to 19931103 (NODC Accession 9400051)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — The Conductivity, Temperature and Depth (CTD) and other data were collected in Bering Sea and North Pacific Ocean. Data was collected from Ship ALPHA HELIX cruise...

Structural basis for the appearance of a molten globule state in chimeric molecules derived from lysozyme and alpha-lactalbumin.

Science.gov (United States)

Joniau, M; Haezebrouck, P; Noyelle, K; Van Dael, H

2001-07-01

The problem as to why alpha-lactalbumin, in the absence of Ca(2+), forms a molten globule intermediate, in contrast to its structural homologue lysozyme, has been addressed by the construction of chimeras of human lysozyme in which either the Ca(2+)-binding loop or a part of helix C of bovine alpha-lactalbumin were transplanted. Previously, we have shown that the introduction of both structural elements together in the lysozyme matrix causes the apo form of the resulting chimera to display molten globule behavior during the course of thermal denaturation. In this article, we demonstrate that this molten globule character is not correlated with the Ca(2+)-binding loop. Also, the Del 101 mutant in which Arg101 was deleted to simulate the alpha-lactalbumin conformation of the connecting loop between helix C and helix D, does not show a stable equilibrium intermediate. Rather, the molten globule character of the chimeras has to be related with a specific part of helix C. More particularly, attention is drawn to the four hydrophobic side-chains I93, V96, I99, and L100, the lysozyme counterparts of which are constituted of less bulky valines and alanine. Our observations are discussed in terms of decreased stability of the native form and increased stability of the intermediate molten globule. Copyright 2001 Wiley-Liss, Inc.

Transport of the alpha-amino-mono-carboxylic acid L-alanine by the beta-alanine carrier of the rabbit ileum

DEFF Research Database (Denmark)

Andersen, Vibeke; Munck, B G

1987-01-01

The proposal that the beta-alanine carrier of the rabbit ileum is a high affinity carrier of the neutral amino acids was examined by means of measurements of influx across the brush border membrane of the intact epithelium using L-alanine as a representative of the neutral amino acids. Confirming...... the proposal, evidence was provided for mutual competitive inhibition between beta-alanine and L-alanine; and it was also demonstrated that a process contributes to the influx of L-alanine, which is characterized by a maximum rate of transport equal to that of beta-alanine and a Kt, which is equal to the Ki...... of L-alanine against the influx of beta-alanine. In the concentration range 0.01 to 0.125 mM the influx of L-alanine was found to be linearly related to the concentration indicating a significant unstirred layer influence on present and previous estimates of the Kt values for influx of amino acids...

Oceanographic profile Biomass, temperature salinity and other measurements collected using bottle from Alpha Helix in the Pacific Ocean from 1976 (NODC Accession 0002070)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — Temperature profile, nutrients, and meteorological data were collected using bottle casts from the ALPHA HELIX in the Pacific Ocean. Data were collected from 06...

An amphipathic alpha-helix controls multiple roles of brome mosaic virus protein 1a in RNA replication complex assembly and function.

Directory of Open Access Journals (Sweden)

Ling Liu

2009-03-01

Full Text Available Brome mosaic virus (BMV protein 1a has multiple key roles in viral RNA replication. 1a localizes to perinuclear endoplasmic reticulum (ER membranes as a peripheral membrane protein, induces ER membrane invaginations in which RNA replication complexes form, and recruits and stabilizes BMV 2a polymerase (2a(Pol and RNA replication templates at these sites to establish active replication complexes. During replication, 1a provides RNA capping, NTPase and possibly RNA helicase functions. Here we identify in BMV 1a an amphipathic alpha-helix, helix A, and use NMR analysis to define its structure and propensity to insert in hydrophobic membrane-mimicking micelles. We show that helix A is essential for efficient 1a-ER membrane association and normal perinuclear ER localization, and that deletion or mutation of helix A abolishes RNA replication. Strikingly, mutations in helix A give rise to two dramatically opposite 1a function phenotypes, implying that helix A acts as a molecular switch regulating the intricate balance between separable 1a functions. One class of helix A deletions and amino acid substitutions markedly inhibits 1a-membrane association and abolishes ER membrane invagination, viral RNA template recruitment, and replication, but doubles the 1a-mediated increase in 2a(Pol accumulation. The second class of helix A mutations not only maintains efficient 1a-membrane association but also amplifies the number of 1a-induced membrane invaginations 5- to 8-fold and enhances viral RNA template recruitment, while failing to stimulate 2a(Pol accumulation. The results provide new insights into the pathways of RNA replication complex assembly and show that helix A is critical for assembly and function of the viral RNA replication complex, including its central role in targeting replication components and controlling modes of 1a action.

Nutrients data collected from the ALPHA HELIX from the coastal waters of Alaska from 03 March 1998 to 07 December 1999 (NODC Accession 0000637)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — Nutrients data were collected from bottle casts from the ALPHA HELIX in the coastal waters of Alaska from 08 March 1998 to 07 December 1999. Data include...

Preparation of new series of poly(amide-imide) reinforced layer silicate nano composite containing N-trimellitimide-L-alanine

International Nuclear Information System (INIS)

Faghihi, K.; Soleimani, M.; Shabanian, M.

2011-01-01

A new poly(amide-imide)-montmorillonite series were generated through solution intercalation technique. Cloisite 20A was used as a modified montmorillonite for ample compatibility with the poly(amide-imide) (PAI) matrix. The PAI 5 chains were synthesized by the direct polycondensation reaction of N-trimellitylimido-L-alanine (3) with 4,4'-diamino diphenyl ether (4) in the presence of tryphenyl phosphites (TPP), CaCl 2 , pyridine and N-methyl-2-pyrrolidone (NMP). Morphology and structure of the resulting PAI-nano composite films 5a-5d with (5-20 Wt%) silicate particles were characterized by Ftir spectroscopy, X-ray diffraction and scanning electron microscopy. The effect of clay dispersion and the interaction between clay and polymeric chains on the properties of nano composites films were investigated by using UV-Vis spectroscopy, thermogravimetric analysis and water uptake measurements. (Author)

Preparation of new series of poly(amide-imide) reinforced layer silicate nano composite containing N-trimellitimide-L-alanine

Energy Technology Data Exchange (ETDEWEB)

Faghihi, K.; Soleimani, M. [Polymer Research Laboratory, Department of Chemistry, Faculty of Science, Islamic Azad University, Arak Branch, Arak (Iran, Islamic Republic of); Shabanian, M., E-mail: k-faghihi@araku.ac.ir [Young Researches Club, Islamic Azad University, Arak Branch, Arak (Iran, Islamic Republic of)

2011-07-01

A new poly(amide-imide)-montmorillonite series were generated through solution intercalation technique. Cloisite 20A was used as a modified montmorillonite for ample compatibility with the poly(amide-imide) (PAI) matrix. The PAI 5 chains were synthesized by the direct polycondensation reaction of N-trimellitylimido-L-alanine (3) with 4,4'-diamino diphenyl ether (4) in the presence of tryphenyl phosphites (TPP), CaCl{sub 2}, pyridine and N-methyl-2-pyrrolidone (NMP). Morphology and structure of the resulting PAI-nano composite films 5a-5d with (5-20 Wt%) silicate particles were characterized by Ftir spectroscopy, X-ray diffraction and scanning electron microscopy. The effect of clay dispersion and the interaction between clay and polymeric chains on the properties of nano composites films were investigated by using UV-Vis spectroscopy, thermogravimetric analysis and water uptake measurements. (Author)

Cloud amount/frequency, NITRATE and other data from ALPHA HELIX in the Chukchi Sea from 1992-09-21 to 1992-10-04 (NODC Accession 9300097)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — The Conductivity, Temperature and Depth (CTD) and other data were collected in Chukchi Sea. Data was collected from Ship ALPHA HELIX. The data was collected over a...

Cloud amount/frequency, NITRATE and other data from ALPHA HELIX in the Bering Sea from 1993-06-12 to 1993-07-01 (NODC Accession 9400026)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — The Conductivity, Temperature and Depth (CTD) and other data were collected in Bering Sea. Data was collected from Ship ALPHA HELIX cruise HX 171. The data was...

In vivo dosimetry with L-alpha-alanine

Energy Technology Data Exchange (ETDEWEB)

Boey, R; Van Der Velden, K [Industriele Hogeschool van het Gemeenschapsonderwijs Limburg, Hasselt (Belgium); Schaeken, B [Algemeen Ziekenhuis Middelheim, Antwerp (Belgium). Dept. of Radiotherapy

1995-12-01

When organic substances are irradiated, stable electrons can be formed. The concentration of these electrons is detected via electron paramagnetic resonance (EPR), a non-destructive form of dosimetry. L-alpha-alanine is extremely suited as a detector because of its high stability and high yield of unpaired electrons. With an EMS 104 spectrometer, we measure the peak-to-peak value of the first derivate of the resonance-spectrum. This value is proportional to the concentration of unpaired electrons and therefore with the absorbed dose. Prior to the in vivo measurements in teletherapy, a calibration curve had to be established. This clearly showed a linear relationship between the EPR-signal and the absorbed dose, except for very low dose where precision was low (20% 1 sd). This indicates that the background signal of the dosimeter is strongly orientation dependent. For this reason it was decided to use pre-irradiated detectors. A number of in vivo measurements has been performed. It was found that the error propagation plays a major role in the calculation of the measured absorbed dose, in the range 1 Gy-6 Gy. Contrary to in vivo measurements in brachytherapy, where higher doses are measured, large uncertainties (30% 1 sd) on the entry dose calculations were observed. For this reason, it is recommended to use a statistical method of reducing this standard deviation to an acceptable level. The proposed method, consisting of 2 detectors and the usage of weight coefficients on our standard deviations, gave promising results. However, theoretical calculations and in vivo measurements show that this method is still not satisfactory to reduce the uncertainty to an acceptable standard in clinical situations.

Chemical and depth data from the ALPHA HELIX using bottle casts in the Bering Sea from 1987-09-07 to 1988-06-11 (NODC Accession 0000263)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — Chemical and depth data were collected from the ALPHA HELIX from September 7, 1987 to June 11, 1988. Data were submitted by the University of Alaska - Fairbanks;...

Cloud amount/frequency, NITRATE and other data from ALPHA HELIX in the Gulf of Alaska from 1990-09-11 to 1990-12-16 (NODC Accession 9100042)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — The Conductivity, Temperature and Depth (CTD) data were collected from Gulf of Alaska using Ship ALPHA HELIX during three cruises HX14-HX145. The data was collected...

Cloud amount/frequency, NITRATE and other data from ALPHA HELIX in the Gulf of Alaska from 1988-09-14 to 1988-09-29 (NODC Accession 8800279)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — The University of Alaska, Institute of Marine Science is responsible for this data collected aboard the R/V Alpha Helix on cruise number HX118 between September 14,...

Temperature and salinity profiles from CTD casts from ALPHA HELIX from NE Pacific (limit-180) from 09 February 1991 to 25 February 1991 (NODC Accession 9100097)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — CTD data was collected from the R/V ALPHA HELIX from the NE Pacific (limit-180). Data were collected by the University of Alaska - Fairbanks; Institute of Marine...

BAROMETRIC PRESSURE and Other Data from ALPHA HELIX From Prince William Sound (Gulf of Alaska) from 1989-05-05 to 1989-05-11 (NODC Accession 8900192)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — The University of Alaska, Institute of Marine Science is responsible for this data collected aboard the R/V Alpha Helix on cruise number HX123 between May 5, 1989 to...

Synthesis, characterization and photo behavior of new poly(amide-imide/montmorillonite nanocomposite containing N,N'-pyrromellitoyl-bis-L-alanine

Directory of Open Access Journals (Sweden)

M. Hajibeygi

2013-09-01

Full Text Available Two new samples of poly(amide-imide-nanocomposites were synthesized by insertion nano silicate particles in poly(amide-imide (PAI chains using a convenient solution intercalation technique. PAI as a source of polymer matrix was synthesized by the direct polycondensation reaction of N,N'-pyrromelitoyl-bis-L-alanine with 4,4'-diamino diphenyl ether in the presence of triphenyl phosphite (TPP, CaCl2, pyridine and N-methyl-2-pyrrolidone (NMP. Morphology and structure of the resulting PAI-nanocomposite films with 5 and 10% silicate particles were characterized by FTIR spectroscopy, X-ray diffraction (XRD and scanning electron microscopy (SEM. The effect of clay dispersion and the interaction between clay and polymeric chains on the properties of nanocomposites films were investigated by using UV-Vis spectroscopy, thermogravimetric analysis (TGA and water uptake measurements.DOI: http://dx.doi.org/10.4314/bcse.v27i3.15

Forced evolution of a regulatory RNA helix in the HIV-1 genome

NARCIS (Netherlands)

Berkhout, B.; Klaver, B.; Das, A. T.

1997-01-01

The 5'and 3'end of the HIV-1 RNA genome forms a repeat (R) element that encodes a double stem-loop structure (the TAR and polyA hairpins). Phylogenetic analysis of the polyA hairpin in different human and simian immunodeficiency viruses suggests that the thermodynamic stability of the helix is

Solitons in an isolated helix chain

DEFF Research Database (Denmark)

Christiansen, Peter Leth; Zolotaryuk, Alexander; Savin, A.V.

1997-01-01

as a generalization of the well-known one-dimensional Fermi-Pasta-Ulam model to include transverse degrees of freedom of the chain molecules. In the particular case of the alpha-helix molecular chain, the intermolecular interactions involved into the model are the point-point bonds connecting the first-, second...

The N-terminal tail of hERG contains an amphipathic α-helix that regulates channel deactivation.

Directory of Open Access Journals (Sweden)

Chai Ann Ng

Full Text Available The cytoplasmic N-terminal domain of the human ether-a-go-go related gene (hERG K+ channel is critical for the slow deactivation kinetics of the channel. However, the mechanism(s by which the N-terminal domain regulates deactivation remains to be determined. Here we show that the solution NMR structure of the N-terminal 135 residues of hERG contains a previously described Per-Arnt-Sim (PAS domain (residues 26-135 as well as an amphipathic α-helix (residues 13-23 and an initial unstructured segment (residues 2-9. Deletion of residues 2-25, only the unstructured segment (residues 2-9 or replacement of the α-helix with a flexible linker all result in enhanced rates of deactivation. Thus, both the initial flexible segment and the α-helix are required but neither is sufficient to confer slow deactivation kinetics. Alanine scanning mutagenesis identified R5 and G6 in the initial flexible segment as critical for slow deactivation. Alanine mutants in the helical region had less dramatic phenotypes. We propose that the PAS domain is bound close to the central core of the channel and that the N-terminal α-helix ensures that the flexible tail is correctly orientated for interaction with the activation gating machinery to stabilize the open state of the channel.

Cloud amount/frequency, NITRATE and other data from ALPHA HELIX in the Bering Sea and Chukchi Sea from 1993-09-09 to 1993-10-10 (NODC Accession 9400036)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — The Conductivity, Temperature and Depth (CTD) and other data were collected in Bering Sea and Chukchi Sea. Data was collected from Ship ALPHA HELIX. The data was...

Cloud amount/frequency, NITRATE and other data from ALPHA HELIX in the Coastal Waters of S. Alaska from 1991-07-24 to 1991-11-14 (NODC Accession 9200039)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — The Conductivity, Temperature and Depth (CTD) and other data were collected in Coastal Waters of S. Alaska. Data was collected in five cruises from Ship ALPHA HELIX....

Physical profile data from CTD casts in the Gulf of Alaska from the R/V ALPHA HELIX from 2001-04-18 to 2001-12-11 (NODC Accession 0000739)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — Physical profile data were collected from CTD casts in the Gulf of Alaska from the R/V Alpha Helix. Data were collected by Western Washington University (WWU) and...

Turbidity, cloud amount/frequency and other data from ALPHA HELIX in the Gulf of Alaska and Bering Sea from 1989-10-04 to 1990-06-26 (NODC Accession 9100032)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — The Conductivity, Temperature and Depth (CTD) and Benthic data were collected from Gulf of Alaska and Bering Sea using NOAA Ship Alpha Helix. The data was collected...

Cloud amount/frequency, TRANSMISSIVITY and other data from ALPHA HELIX and LITTLE DIPPER in the Gulf of Alaska from 1992-01-28 to 1992-04-22 (NODC Accession 9200146)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — The Conductivity, Temperature and Depth (CTD) and other data were collected in Gulf of Alaska. Data was collected from Ships ALPHA HELIX and LITTLE DIPPER. The data...

Improved synthesis with high yield and increased molecular weight of poly(alpha,beta-malic acid) by direct polycondensation.

Science.gov (United States)

Kajiyama, Tetsuto; Kobayashi, Hisatoshi; Taguchi, Tetsushi; Kataoka, Kazunori; Tanaka, Junzo

2004-01-01

The development of synthetic biodegradable polymers, such as poly(alpha-hydroxy acid), is particularly important for constructing medical devices, including scaffolds and sutures, and has attracted growing interest in the biomedical field. Here, we report a novel approach to preparing high molecular weight poly(malic acid) (HMW--PMA) as a biodegradable and bioabsorbable water-soluble polymer. We investigated in detail the reaction conditions for the simple direct polycondensation of l-malic acid, including the reaction times, temperatures, and catalysts. The molecular weight of synthesized alpha,beta-PMA is dependent on both the reaction temperature and time. The optimum reaction condition to obtain alpha,beta-PMA by direct polycondensation using tin(II) chloride as a catalyst was thus determined to be 110 degrees C for 45 h with a molecular weight of 5300. The method for alpha,beta-PMA synthesis established here will facilitate production of alpha,beta-PMA of various molecular weights, which may have a potential utility as biomaterials.

Physical and nutrient profile data from bottle casts from the R/V ALPHA HELIX in Gulf of Alaska from 03 December 1990 to 11 December 1998 (NODC Accession 0000221)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — Physical and nutrients data were collected using bottle casts from the R/V ALPHA HELIX in Gulf of Alaska from December 3, 1980 to December 11, 1998. These data were...

A Dual-Purpose Linker for Alpha Helix Stabilization and Imaging Agent Conjugation to Glucagon-Like Peptide-1 Receptor Ligands

Science.gov (United States)

Zhang, Liang; Navaratna, Tejas; Liao, Jianshan; Thurber, Greg M.

2016-01-01

Peptides display many characteristics of efficient imaging agents such as rapid targeting, fast background clearance, and low non-specific cellular uptake. However, poor stability, low affinity, and loss of binding after labeling often preclude their use in vivo. Using the glucagon-like peptide-1 receptor (GLP-1R) ligands exendin and GLP-1 as a model system, we designed a novel alpha helix stabilizing linker to simultaneously address these limitations. The stabilized and labeled peptides showed an increase in helicity, improved protease resistance, negligible loss or an improvement in binding affinity, and excellent in vivo targeting. The ease of incorporating azidohomoalanine in peptides and efficient reaction with the dialkyne linker enables this technique to potentially be used as a general method for labeling alpha helices. This strategy should be useful for imaging beta cells in diabetes research and in developing and testing other peptide targeting agents. PMID:25594741

A comparison of three materials used in ESR dosimetry: L-α-alanine, DL-α-alanine and standard bone powder. Response to Co-60 gamma radiation

International Nuclear Information System (INIS)

Stuglik, Z.; Sadlo, J.

1995-01-01

Three solid state materials: L-α-alanine, DL-α-alanine and standard bone powder were irradiated with gamma analyzed with ESR method. It was stated that the G-value of paramagnetic centres in L-α-alanine is practically the same as in DL-alpha-alanine and about 50 times higher than in non-deproteinized bone powder. The sensitivities of investigated materials are proportional to their G-values if double integrals of ESR signals are chosen as a measure of radiation effects. When first derivatives of ESR absorption bands are used to the construction of dose-response curves (peak-to-peak method) the sensitivities of all investigated materials are comparable. (author). 14 refs, 1 fig., 3 tabs

Physical and nutrients profile data from the R/V ALPHA HELIX using bottle casts in the Gulf of Alaska from 06 April 1989 to 11 April 1989 (NODC Accession 0000223)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — Physical and nutrient profile data were collected using bottle casts from the R/V ALPHA HELIX in the Gulf of Alaska from April 6, 1989 to April 11, 1989. Data were...

A network of hydrophobic residues impeding helix alphaC rotation maintains latency of kinase Gcn2, which phosphorylates the alpha subunit of translation initiation factor 2.

Science.gov (United States)

Gárriz, Andrés; Qiu, Hongfang; Dey, Madhusudan; Seo, Eun-Joo; Dever, Thomas E; Hinnebusch, Alan G

2009-03-01

Kinase Gcn2 is activated by amino acid starvation and downregulates translation initiation by phosphorylating the alpha subunit of translation initiation factor 2 (eIF2alpha). The Gcn2 kinase domain (KD) is inert and must be activated by tRNA binding to the adjacent regulatory domain. Previous work indicated that Saccharomyces cerevisiae Gcn2 latency results from inflexibility of the hinge connecting the N and C lobes and a partially obstructed ATP-binding site in the KD. Here, we provide strong evidence that a network of hydrophobic interactions centered on Leu-856 also promotes latency by constraining helix alphaC rotation in the KD in a manner relieved during amino acid starvation by tRNA binding and autophosphorylation of Thr-882 in the activation loop. Thus, we show that mutationally disrupting the hydrophobic network in various ways constitutively activates eIF2alpha phosphorylation in vivo and bypasses the requirement for a key tRNA binding motif (m2) and Thr-882 in Gcn2. In particular, replacing Leu-856 with any nonhydrophobic residue activates Gcn2, while substitutions with various hydrophobic residues maintain kinase latency. We further provide strong evidence that parallel, back-to-back dimerization of the KD is a step on the Gcn2 activation pathway promoted by tRNA binding and autophosphorylation. Remarkably, mutations that disrupt the L856 hydrophobic network or enhance hinge flexibility eliminate the need for the conserved salt bridge at the parallel dimer interface, implying that KD dimerization facilitates the reorientation of alphaC and remodeling of the active site for enhanced ATP binding and catalysis. We propose that hinge remodeling, parallel dimerization, and reorientation of alphaC are mutually reinforcing conformational transitions stimulated by tRNA binding and secured by the ensuing autophosphorylation of T882 for stable kinase activation.

Physical profile data from the R/V ALPHA HELIX in the Gulf of Alaska as part of the GLOBEC Project from 02 October 1998 to 07 December 1999 (NODC Accession 0000238)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — Physical profile data were collected from the R/V ALPHA HELIX in the Gulf of Alaska from October 2, 1998 to December 7, 1999. Data were submitted by the University...

Living Polymerization of N -Substituted β-Alanine N -Carboxyanhydrides: Kinetic Investigations and Preparation of an Amphiphilic Block Copoly-β-Peptoid

KAUST Repository

Grossmann, Arlett

2012-07-03

Poly(α-peptoid)s (N-substituted polyglycines) are interesting peptidomimetic biomaterials that have been discussed for many applications. Poly(β-peptoid)s (N-substituted poly-β-alanines), although equally intriguing, have received much less attention. Here we present results that suggest that while N-substituted β-alanine N-carboxyanhydrides can undergo a living nucleophilic ring-opening polymerization, the solubility of poly(β-peptoid)s can be very poor, which contributes to the limited accessibility using other synthetic approaches. The living character of the polymerization was utilized for the preparation of the first polymerized amphiphilic block copoly-β-peptoid. Our results may open a new route towards highly defined functional poly(β-peptoid)s which could represent biomaterials. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

MicroRNA-212 post-transcriptionally regulates oocyte-specific basic-helix-loop-helix transcription factor, factor in the germline alpha (FIGLA, during bovine early embryogenesis.

Directory of Open Access Journals (Sweden)

Swamy K Tripurani

Full Text Available Factor in the germline alpha (FIGLA is an oocyte-specific basic helix-loop-helix transcription factor essential for primordial follicle formation and expression of many genes required for folliculogenesis, fertilization and early embryonic survival. Here we report the characterization of bovine FIGLA gene and its regulation during early embryogenesis. Bovine FIGLA mRNA expression is restricted to gonads and is detected in fetal ovaries harvested as early as 90 days of gestation. FIGLA mRNA and protein are abundant in germinal vesicle and metaphase II stage oocytes, as well as in embryos from pronuclear to eight-cell stage but barely detectable at morula and blastocyst stages, suggesting that FIGLA might be a maternal effect gene. Recent studies in zebrafish and mice have highlighted the importance of non-coding small RNAs (microRNAs as key regulatory molecules targeting maternal mRNAs for degradation during embryonic development. We hypothesized that FIGLA, as a maternal transcript, is regulated by microRNAs during early embryogenesis. Computational predictions identified a potential microRNA recognition element (MRE for miR-212 in the 3' UTR of the bovine FIGLA mRNA. Bovine miR-212 is expressed in oocytes and tends to increase in four-cell and eight-cell stage embryos followed by a decline at morula and blastocyst stages. Transient transfection and reporter assays revealed that miR-212 represses the expression of FIGLA in a MRE dependent manner. In addition, ectopic expression of miR-212 mimic in bovine early embryos dramatically reduced the expression of FIGLA protein. Collectively, our results demonstrate that FIGLA is temporally regulated during bovine early embryogenesis and miR-212 is an important negative regulator of FIGLA during the maternal to zygotic transition in bovine embryos.

Effects of hydrophobic helix length and side chain chemistry on biomimicry in peptoid analogues of SP-C.

Science.gov (United States)

Brown, Nathan J; Wu, Cindy W; Seurynck-Servoss, Shannon L; Barron, Annelise E

2008-02-12

The hydrophobic proteins of lung surfactant (LS), SP-B and SP-C, are critical constituents of an effective surfactant replacement therapy for the treatment of respiratory distress syndrome. Because of concerns and difficulties associated with animal-derived surfactants, recent investigations have focused on the creation of synthetic analogues of the LS proteins. However, creating an accurate mimic of SP-C that retains its biophysical surface activity is extraordinarily challenging given the lipopeptide's extreme hydrophobicity and propensity to misfold and aggregate. One successful approach that overcomes these difficulties is the use of poly-N-substituted glycines, or peptoids, to mimic SP-C. To develop a non-natural, bioactive mimic of SP-C and to investigate the effects of side chain chemistry and length of the helical hydrophobic region, we synthesized, purified, and performed in vitro testing of two classes of peptoid SP-C mimics: those having a rigid alpha-chiral aromatic helix and those having a biomimetic alpha-chiral aliphatic helix. The length of the two classes of mimics was also systematically altered. Circular dichroism spectroscopy gave evidence that all of the peptoid-based mimics studied here emulated SP-C's secondary structure, forming stable helical structures in solution. Langmuir-Wilhelmy surface balance, fluorescence microscopy, and pulsating bubble surfactometry experiments provide evidence that the aromatic-based SP-C peptoid mimics, in conjunction with a synthetic lipid mixture, have superior surface activity and biomimetic film morphology in comparison to the aliphatic-based mimics and that there is an increase in surface activity corresponding to increasing helical length.

Helix-helix inversion of an optically-inactive π-conjugated foldamer triggered by concentration changes of a single enantiomeric guest leading to a change in the helical stability.

Science.gov (United States)

Liu, Lijia; Ousaka, Naoki; Horie, Miki; Mamiya, Fumihiko; Yashima, Eiji

2016-09-27

A preferred-handed helicity induced in an optically-inactive poly(phenyleneethynylene)-based foldamer bearing carboxylic acid pendants upon complexation with a single enantiomeric diamine was subsequently inverted into the opposite helix upon further addition of the diamine, accompanied by a remarkable change in the stability of the helices.

Profile nutrients data from bottle casts from the R/V Alpha Helix in the Bering Sea, Gulf of Alaska and Northeast Pacific from 16 August 1983 to 30 July 1989 (NODC Accession 0000252)

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National Oceanic and Atmospheric Administration, Department of Commerce — Profile nutrients and other data were collected from the R/V ALPHA HELIX from August 16, 1983 to July 30, 1989. Data were submitted by University of Alaska -...

Nutrients data from the R/V ALPHA HELIX and T.G. THOMPSON as part of the Inner Shelf Transfer and Recycling Project from 17 April 1987 to 21 July 1988 (NODC Accession 0000296)

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National Oceanic and Atmospheric Administration, Department of Commerce — Nitrate, nitrite, phosphate, silicate, and other data were collected from the R/V ALPHA HELIX and THOMAS G. THOMPSON from April 17, 1987 to July 21, 1988. Data were...

General architecture of the alpha-helical globule.

Science.gov (United States)

Murzin, A G; Finkelstein, A V

1988-12-05

A model is presented for the arrangement of alpha-helices in globular proteins. In the model, helices are placed on certain ribs of "quasi-spherical" polyhedra. The polyhedra are chosen so as to allow the close packing of helices around a hydrophobic core and to stress the collective interactions of the individual helices. The model predicts a small set of stable architectures for alpha-helices in globular proteins and describes the geometries of the helix packings. Some of the predicted helix arrangements have already been observed in known protein structures; others are new. An analysis of the three-dimensional structures of all proteins for which co-ordinates are available shows that the model closely approximates the arrangements and packing of helices actually observed. The average deviations of the real helix axes from those in the model polyhedra is +/- 20 degrees in orientation and +/- 2 A in position (1 A = 0.1 nm). We also show that for proteins that are not homologous, but whose helix arrangements are described by the same polyhedron, the root-mean-square difference in the position of the C alpha atoms in the helices is 1.6 to 3.0 A.

No need to be HAMLET or BAMLET to interact with histones: binding of monomeric alpha-lactalbumin to histones and basic poly-amino acids.

Science.gov (United States)

Permyakov, Serge E; Pershikova, Irina V; Khokhlova, Tatyana I; Uversky, Vladimir N; Permyakov, Eugene A

2004-05-18

The ability of a specific complex of human alpha-lactalbumin with oleic acid (HAMLET) to induce cell death with selectivity for tumor and undifferentiated cells was shown recently to be mediated by interaction of HAMLET with histone proteins irreversibly disrupting chromatin structure [Duringer, C., et al. (2003) J. Biol. Chem. 278, 42131-42135]. Here we show that monomeric alpha-lactalbumin (alpha-LA) in the absence of fatty acids is also able to bind efficiently to the primary target of HAMLET, histone HIII, regardless of Ca(2+) content. Thus, the modification of alpha-LA by oleic acid is not required for binding to histones. We suggest that interaction of negatively charged alpha-LA with the basic histone stabilizes apo-alpha-LA and destabilizes the Ca(2+)-bound protein due to compensation for excess negative charge of alpha-LA's Ca(2+)-binding loop by positively charged residues of the histone. Spectrofluorimetric curves of titration of alpha-LA by histone H3 were well approximated by a scheme of cooperative binding of four alpha-LA molecules per molecule of histone, with an equilibrium dissociation constant of 1.0 microM. Such a stoichiometry of binding implies that the binding process is not site-specific with respect to histone and likely is driven by just electrostatic interactions. Co-incubation of positively charged poly-amino acids (poly-Lys and poly-Arg) with alpha-LA resulted in effects which were similar to those caused by histone HIII, confirming the electrostatic nature of the alpha-LA-histone interaction. In all cases that were studied, the binding was accompanied by aggregation. The data indicate that alpha-lactalbumin can be used as a basis for the design of antitumor agents, acting through disorganization of chromatin structure due to interaction between alpha-LA and histone proteins.

Synthesis and Analysis of Methacryloyl-L-Alanine Methyl Ester using fourier Transform Nuclear Magnetic Resonance

International Nuclear Information System (INIS)

Tri Darwinto

2008-01-01

Methacryloyl-L-alanine methyl ester was synthesized by reacting methacrylic acid with L-alanine methyl ester hydrochloride in triethylamine at temperature of 90 o C. Hydrogel polymer of poly(methacryloyl-L-alanine methyl ester) was much used for diagnosis and therapy of vascular tumor. The molecular structure methacryloyl-L-alanine methyl ester analyzed by fourier transform nuclear magnetic resonance (FT-NMR) for analyzing of carbon atom ( 13 C) using Distortionless Enhancement by Polarization Transfer (DEPT) measurement mode with coupling as well as without coupling from proton atom ( 1 H). Molecular structure analysis result showed that DEPT FT-NMR measurement mode with coupling as well as without coupling from 1 H was very fast, exact and accurate method for molecular analysis of organic compound especially methacryloyl-L-alanine methyl ester. (author)

Nutrients data from bottle casts in the Bering Sea from the R/V ALPHA HELIX as part of the Inner Shelf Transport and Recycling Project 04 July 1986 to 25 September 1986 (NODC Accession 0000293)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — Nitrate, nitrite, phosphate, silicate, and other data were collected from the R/V ALPHA HELIX from July 4, 1986 to September 25, 1986. Data were submitted by...

Current components, physical, and other data from moored current meters and CTD casts from the ALPHA HELIX and other platforms from Gulf of Alaska from 1988-04-28 to 1988-06-29 (NODC Accession 8800237)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — Current components, physical, and other data were collected by moored current meters and CTD casts in the Gulf of Alaska from the ALPHA HELIX and other platforms....

Nutrients profile data from the R/V ALPHA HELIX in the Bering Sea in support of the Inner Shelf Transport and Recycling Project (ISHTAR) from 13 July 1985 to 14 October 1985 (NODC Accession 0000276)

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National Oceanic and Atmospheric Administration, Department of Commerce — Ammonium, nitrate, nitrite, phosphate, silicate, and urea data were collected from the R/V ALPHA HELIX from July 13, 1985 to October 14, 1985. Data were submitted by...

Experiences with alanine dosimetry in afterloading brachytherapy

International Nuclear Information System (INIS)

Eberhardt, H.-J.; Gohs, U.

1996-01-01

At the present, the most commonly used dosimetry for radiotherapy applications are ionisation chambers and thermoluminescent dosimeters (TLD). However, there are some undesirable characteristics of these dosimetry systems, such as large detection volume (ionisation chamber) as well as fading of the radiation induced signal with time and destructive readout (TLG). The present study is an investigation into the use of the alanine/ESR dosimetry in fractionated afterloading brachytherapy during the whole radiotherapy course. There are some qualities which make alanine dosimetry attractive. These are the linear energy response, low fading under standard conditions, and the nondestructive readout. Thus the alanine dosimetry makes possible cumulative dose measurements during the radiotherapy course and an archival storage. By ionizing radiation (gamma, e, n, p, charged particles) free radicals (unpaired electrons) are produced in the amino acid alanine. The continuous wave electron spin resonance (ESR) spectroscopy is used to determine the number of free radicals, which is proportional to the absorbed dose and the alanine content of the dosimeter. The ESR measurements were made at room temperature using a Bruker EPR analyzer EMS-104. The dosimeters used in the test are alanine pellets (23.72 mg weight, 4.9 mm diameter, 1 mm height) as well as flexible alanine film dosimeters (thickness about 500 μm). The dosimeters consist of a blend of L-alpha-alanine and a binder. The alanine content of the pellets and the film dosimeters is about 88 % and 50 % by weight, respectively. The dosimeters for the calculation of the dose-effect-relationship were irradiated at the Physical-Technical Bundesanstalt in Braunschweig by a standard 60Co source. The maximum deviation from the calculated linear function is about 0.12 Gy in the dose range up to 80 Gy. The goal of medical applications was the superficial dose measurement in afterloading brachytherapy during the radiotherapy course in

Titration of Alanine Monitored by NMR Spectroscopy: A Biochemistry Laboratory Experiment

Science.gov (United States)

Waller, Francis J.; And Others

1977-01-01

The experiment described here involves simultaneous monitoring of pH and NMR chemical shifts during an aqueous titration of alpha- and beta-alanine. This experiment is designed for use in an undergraduate biochemistry course. (MR)

CTD, current meter, meteorological buoy, and bottle data from the Gulf of Mexico from the ALPHA HELIX and other platforms in support of LATEX A from 18 March 1993 to 23 September 1993 (NODC Accession 9400149)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — CTD, current meter, meteorological buoy, and bottle data were collected from the Gulf of Mexico from the ALPHA HELIX and other platforms. Data were collected by...

Catalytic Stereoinversion of L-Alanine to Deuterated D-Alanine.

Science.gov (United States)

Moozeh, Kimia; So, Soon Mog; Chin, Jik

2015-08-03

A combination of an achiral pyridoxal analogue and a chiral base has been developed for catalytic deuteration of L-alanine with inversion of stereochemistry to give deuterated D-alanine under mild conditions (neutral pD and 25 °C) without the use of any protecting groups. This system can also be used for catalytic deuteration of D-alanine with retention of stereochemistry to give deuterated D-alanine. Thus a racemic mixture of alanine can be catalytically deuterated to give an enantiomeric excess of deuterated D-alanine. While catalytic deracemization of alanine is forbidden by the second law of thermodynamics, this system can be used for catalytic deracemization of alanine with deuteration. Such green and biomimetic approach to catalytic stereocontrol provides insights into efficient amino acid transformations. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Teaching helix and problems connected with helix using GeoGebra

Science.gov (United States)

Bímová, Daniela

2017-12-01

The contribution presents the dynamic applets created in GeoGebra that show the origin and main properties of a helix and it also presents some constructive problems connected with the helix. There are created the step by step algorithms of some constructions in the chosen applets. Three-dimensional applets include illustrative helix samples and spatial animations that help students better see problems concerning the helix spatially. There is mentioned the website in the contribution on which there is situated GeoGebra book dedicated to the topic "Helix" and containing the mentioned applets. The created applets and materials of the GeoGebra book "Helix" help in teaching and studying the course Constructive Geometry determined for the students of the Faculty of Mechanical Engineering of the Technical University of Liberec.

Protein Secondary Structures (alpha-helix and beta-sheet) at a Cellular Levle and Protein Fractions in Relation to Rumen Degradation Behaviours of Protein: A New Approach

Energy Technology Data Exchange (ETDEWEB)

Yu,P.

2007-01-01

Studying the secondary structure of proteins leads to an understanding of the components that make up a whole protein, and such an understanding of the structure of the whole protein is often vital to understanding its digestive behaviour and nutritive value in animals. The main protein secondary structures are the {alpha}-helix and {beta}-sheet. The percentage of these two structures in protein secondary structures influences protein nutritive value, quality and digestive behaviour. A high percentage of {beta}-sheet structure may partly cause a low access to gastrointestinal digestive enzymes, which results in a low protein value. The objectives of the present study were to use advanced synchrotron-based Fourier transform IR (S-FTIR) microspectroscopy as a new approach to reveal the molecular chemistry of the protein secondary structures of feed tissues affected by heat-processing within intact tissue at a cellular level, and to quantify protein secondary structures using multicomponent peak modelling Gaussian and Lorentzian methods, in relation to protein digestive behaviours and nutritive value in the rumen, which was determined using the Cornell Net Carbohydrate Protein System. The synchrotron-based molecular chemistry research experiment was performed at the National Synchrotron Light Source at Brookhaven National Laboratory, US Department of Energy. The results showed that, with S-FTIR microspectroscopy, the molecular chemistry, ultrastructural chemical make-up and nutritive characteristics could be revealed at a high ultraspatial resolution ({approx}10 {mu}m). S-FTIR microspectroscopy revealed that the secondary structure of protein differed between raw and roasted golden flaxseeds in terms of the percentages and ratio of {alpha}-helixes and {beta}-sheets in the mid-IR range at the cellular level. By using multicomponent peak modelling, the results show that the roasting reduced (P <0.05) the percentage of {alpha}-helixes (from 47.1% to 36.1%: S

Depth, salinity, and other data were collected from the R/V ALPHA HELIX in the Bering Sea as part of the Inner Shelf Transport and Recycling Project (ISHTAR) from 17 September 1986 to 25 September 1986 (NODC Accession 0000275)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — Depth, fluorescence, oxygen, salinity, and water temperature data were collected from the R/V ALPHA HELIX from September 17, 1986 to September 25, 1986. Data were...

Tipping the Scale from Disorder to Alpha-helix: Folding of Amphiphilic Peptides in the Presence of Macroscopic and Molecular Interfaces.

Science.gov (United States)

Dalgicdir, Cahit; Globisch, Christoph; Peter, Christine; Sayar, Mehmet

2015-08-01

Secondary amphiphilicity is inherent to the secondary structural elements of proteins. By forming energetically favorable contacts with each other these amphiphilic building blocks give rise to the formation of a tertiary structure. Small proteins and peptides, on the other hand, are usually too short to form multiple structural elements and cannot stabilize them internally. Therefore, these molecules are often found to be structurally ambiguous up to the point of a large degree of intrinsic disorder in solution. Consequently, their conformational preference is particularly susceptible to environmental conditions such as pH, salts, or presence of interfaces. In this study we use molecular dynamics simulations to analyze the conformational behavior of two synthetic peptides, LKKLLKLLKKLLKL (LK) and EAALAEALAEALAE (EALA), with built-in secondary amphiphilicity upon forming an alpha-helix. We use these model peptides to systematically study their aggregation and the influence of macroscopic and molecular interfaces on their conformational preferences. We show that the peptides are neither random coils in bulk water nor fully formed alpha helices, but adopt multiple conformations and secondary structure elements with short lifetimes. These provide a basis for conformation-selection and population-shift upon environmental changes. Differences in these peptides' response to macroscopic and molecular interfaces (presented by an aggregation partner) can be linked to their inherent alpha-helical tendencies in bulk water. We find that the peptides' aggregation behavior is also strongly affected by presence or absence of an interface, and rather subtly depends on their surface charge and hydrophobicity.

Glucose-stimulated prehepatic insulin secretion is associated with circulating alanine, triglyceride, glucagon, lactate and TNF-alpha in patients with HIV-lipodystrophy

DEFF Research Database (Denmark)

Haugaard, S B; Andersen, O; Pedersen, S B

2006-01-01

with the remaining HIV-infected patients (all Ptriglyceride, alanine, glucagon, lactate and TNF-alpha may be associated with alterations in the first-phase prehepatic insulin secretion response to intravenous glucose in normoglycaemic lipodystrophic HIV-infected patients.......OBJECTIVES: We examined whether insulin-resistant lipodystrophic HIV-infected patients with known high fasting prehepatic insulin secretion rates (FISRs) displayed alterations in first-phase prehepatic insulin response to intravenous glucose (ISREG0-10 min). METHODS: Eighteen normoglycaemic...... lipodystrophic HIV-infected (LIPO) patients and 25 normoglycaemic nonlipodystrophic HIV-infected patients (controls) were included in the study. The prehepatic insulin secretion rate was estimated by deconvolution of C-peptide concentrations, and insulin sensitivity (SIRd) was estimated by the glucose clamp...

Lower critical solution temperature behavior of alpha-substituted poly(acrylic acids)s, cyclopolymerization of N-vinylformamido-methylacrylates, and use of the World-Wide Web in polymer science education

Science.gov (United States)

Michalovic, Mark Stephen

A series of alpha-substituted poly(acrylic acid)s was synthesized and characterized. Their aqueous solution properties were investigated with respect to lower critical solution temperature (LCST) behavior. Poly(alpha-methoxymethylacrylic acid) was found to have a lower critical solution temperature (LCST) of 46°C, poly(alpha-methoxyethoxymethylacrylic acid) showed an LCST of 26.5°C and poly(alpha-methoxyethoxyethoxymethylacrylic acid) showed an LCST of 66°C. The cloud points of the solutions of these polymers were found to be sensitive to pH, and to concentrations of additives such as urea, salts, and surfactants. Because of low molecular weight due to chain transfer, high molecular weight analogs of the ether-linked polymers were synthesized in which ester linkages joined the oligo-oxyethylene segment to the acrylate moiety. Poly(alpha-methoxyethoxyacetoxymethylacrylic acid) was the only one of this series to give an LCST with a value of 52.5°C. Copolymers of t-butyl alpha-methoxymethylacrylate (tBMMA) with alpha-(1H,1H- perfluorooctyloxymethyl)acrylic acid (PFOMA) were synthesized, deprotected and their lower critical solution temperatures (LCSTs) evaluated. At PFOMA feed ratios of 0.25 mol % or less, no observable change in the LCST was observed, while at PFOMA feed ratios of above 0.25 mol % to 1.125 mol %, a large linear decrease in the LCST was observed with increasing fluorocarbon content. t-Butyl alpha-(N-vinylformamidomethyl)acrylate (tBVFA) and ethyl alpha-(N-vinylformamidomethyl)acrylate (EVFA) were synthesized from t-butyl alpha-bromomethylacrylate and ethyl alpha-chloromethylacrylate, respectively. tBVFA was found to cyclopolymerize at 120°C in DMF, DMSO, and 1,2-dichlorobenzene at solvent:monomer ratios of 10:1 vol:wt. Molecular weights for poly(tBVFA) ranged from 10,000 to 13,000 as estimated by size-exclusion chromatography. At lower solvent monomer ratio (1:1), and at lower temperature (71°C), crosslinking occurred. EVFA was found to

Optimized molecular dynamics force fields applied to the helix-coil transition of polypeptides.

Science.gov (United States)

Best, Robert B; Hummer, Gerhard

2009-07-02

Obtaining the correct balance of secondary structure propensities is a central priority in protein force-field development. Given that current force fields differ significantly in their alpha-helical propensities, a correction to match experimental results would be highly desirable. We have determined simple backbone energy corrections for two force fields to reproduce the fraction of helix measured in short peptides at 300 K. As validation, we show that the optimized force fields produce results in excellent agreement with nuclear magnetic resonance experiments for folded proteins and short peptides not used in the optimization. However, despite the agreement at ambient conditions, the dependence of the helix content on temperature is too weak, a problem shared with other force fields. A fit of the Lifson-Roig helix-coil theory shows that both the enthalpy and entropy of helix formation are too small: the helix extension parameter w agrees well with experiment, but its entropic and enthalpic components are both only about half the respective experimental estimates. Our structural and thermodynamic analyses point toward the physical origins of these shortcomings in current force fields, and suggest ways to address them in future force-field development.

The unresolved puzzle why alanine extensions cause disease.

Science.gov (United States)

Winter, Reno; Liebold, Jens; Schwarz, Elisabeth

2013-08-01

The prospective increase in life expectancy will be accompanied by a rise in the number of elderly people who suffer from ill health caused by old age. Many diseases caused by aging are protein misfolding diseases. The molecular mechanisms underlying these disorders receive constant scientific interest. In addition to old age, mutations also cause congenital protein misfolding disorders. Chorea Huntington, one of the most well-known examples, is caused by triplet extensions that can lead to more than 100 glutamines in the N-terminal region of huntingtin, accompanied by huntingtin aggregation. So far, nine disease-associated triplet extensions have also been described for alanine codons. The extensions lead primarily to skeletal malformations. Eight of these proteins represent transcription factors, while the nuclear poly-adenylate binding protein 1, PABPN1, is an RNA binding protein. Additional alanines in PABPN1 lead to the disease oculopharyngeal muscular dystrophy (OPMD). The alanine extension affects the N-terminal domain of the protein, which has been shown to lack tertiary contacts. Biochemical analyses of the N-terminal domain revealed an alanine-dependent fibril formation. However, fibril formation of full-length protein did not recapitulate the findings of the N-terminal domain. Fibril formation of intact PABPN1 was independent of the alanine segment, and the fibrils displayed biochemical properties that were completely different from those of the N-terminal domain. Although intranuclear inclusions have been shown to represent the histochemical hallmark of OPMD, their role in pathogenesis is currently unclear. Several cell culture and animal models have been generated to study the molecular processes involved in OPMD. These studies revealed a number of promising future therapeutic strategies that could one day improve the quality of life for the patients.

Tipping the Scale from Disorder to Alpha-helix: Folding of Amphiphilic Peptides in the Presence of Macroscopic and Molecular Interfaces.

Directory of Open Access Journals (Sweden)

Cahit Dalgicdir

2015-08-01

Full Text Available Secondary amphiphilicity is inherent to the secondary structural elements of proteins. By forming energetically favorable contacts with each other these amphiphilic building blocks give rise to the formation of a tertiary structure. Small proteins and peptides, on the other hand, are usually too short to form multiple structural elements and cannot stabilize them internally. Therefore, these molecules are often found to be structurally ambiguous up to the point of a large degree of intrinsic disorder in solution. Consequently, their conformational preference is particularly susceptible to environmental conditions such as pH, salts, or presence of interfaces. In this study we use molecular dynamics simulations to analyze the conformational behavior of two synthetic peptides, LKKLLKLLKKLLKL (LK and EAALAEALAEALAE (EALA, with built-in secondary amphiphilicity upon forming an alpha-helix. We use these model peptides to systematically study their aggregation and the influence of macroscopic and molecular interfaces on their conformational preferences. We show that the peptides are neither random coils in bulk water nor fully formed alpha helices, but adopt multiple conformations and secondary structure elements with short lifetimes. These provide a basis for conformation-selection and population-shift upon environmental changes. Differences in these peptides' response to macroscopic and molecular interfaces (presented by an aggregation partner can be linked to their inherent alpha-helical tendencies in bulk water. We find that the peptides' aggregation behavior is also strongly affected by presence or absence of an interface, and rather subtly depends on their surface charge and hydrophobicity.

Three-residue turns in alpha/beta-peptides and their application in the design of tertiary structures.

Science.gov (United States)

Sharma, Gangavaram V M; Nagendar, Pendem; Ramakrishna, Kallaganti V S; Chandramouli, Nagula; Choudhary, Madavi; Kunwar, Ajit C

2008-06-02

A new three-residue turn was serendipitously discovered in alpha/beta hybrid peptides derived from alternating C-linked carbo-beta-amino acids (beta-Caa) and L-Ala residues. The three-residue beta-alpha-beta turn at the C termini, nucleated by a helix at the N termini, resulted in helix-turn (HT) supersecondary structures in these peptides. The turn in the HT motif is stabilized by two H bonds-CO(i-2)-NH(i), with a seven-membered pseudoring (gamma turn) in the backward direction, and NH(i-2)-CO(i), with a 13-membered pseudoring in the forward direction (i being the last residue)--at the C termini. The study was extended to generalize the new three-residue turn (beta-alpha-beta) by using different alpha- and beta-amino acids. Furthermore, the HT motifs were efficiently converted, by an extension with helical oligomers at the C termini, into peptides with novel helix-turn-helix (HTH) tertiary structures. However, this resulted in the destabilization of the beta-alpha-beta turn with the concomitant nucleation of another three-residue turn, alpha-beta-beta, which is stabilized by 11- and 15-membered bifurcated H bonds. Extensive NMR spectroscopic studies were carried out to delineate the secondary and tertiary structures in these peptides, which are further supported by molecular dynamics (MD) investigations.

Alpha particle response for a prototype radiation survey meter based on poly(ethylene terephthalate) with un-doping fluorescent guest molecules

International Nuclear Information System (INIS)

Nguyen, Philip; Nakamura, Hidehito; Sato, Nobuhiro; Takahashi, Tomoyuki; Maki, Daisuke; Kanayama, Masaya; Takahashi, Sentaro; Kitamura, Hisashi; Shirakawa, Yoshiyuki

2016-01-01

There is no radiation survey meter that can discriminate among alpha particles, beta particles, and gamma-rays with one material. Previously, undoped poly(ethylene terephthalate) (PET) has been shown to be an effective material for beta particle and gamma-ray detection. Here, we demonstrate a prototype survey meter for alpha particles based on undoped PET. A 140 × 72 × 1-mm PET substrate was fabricated with mirrored surfaces. It was incorporated in a unique detection section of the survey meter that directly detects alpha particles. The prototype exhibited an unambiguous response to alpha particles from a 241 Am radioactive source. These results demonstrate that undoped PET can perform well in survey meters for alpha particle detection. Overall, the PET-based survey meter has the potential to detect multiple types of radiation, and will spawn an unprecedented type of radiation survey meter based on undoped aromatic ring polymers. (author)

Alanine water complexes.

Science.gov (United States)

Vaquero, Vanesa; Sanz, M Eugenia; Peña, Isabel; Mata, Santiago; Cabezas, Carlos; López, Juan C; Alonso, José L

2014-04-10

Two complexes of alanine with water, alanine-(H2O)n (n = 1,2), have been generated by laser ablation of the amino acid in a supersonic jet containing water vapor and characterized using Fourier transform microwave spectroscopy. In the observed complexes, water molecules bind to the carboxylic group of alanine acting as both proton donors and acceptors. In alanine-H2O, the water molecule establishes two intermolecular hydrogen bonds forming a six-membered cycle, while in alanine-(H2O)2 the two water molecules establish three hydrogen bonds forming an eight-membered ring. In both complexes, the amino acid moiety is in its neutral form and shows the conformation observed to be the most stable for the bare molecule. The microsolvation study of alanine-(H2O)n (n = 1,2) can be taken as a first step toward understanding bulk properties at a microscopic level.

Theoretical and experimental radiation effectiveness of the free radical dosimeter alanine to irradiation with heavy charged particles

DEFF Research Database (Denmark)

Hansen, Jørgen-Walther; Olsen, K. J.

1985-01-01

Dose-response characteristics have been measured for the crystalline amino acid L-.alpha.-alanine irradiated with ion beams of 6 and 16 MeV protons, 20 MeV .alpha. particles, 21 MeV7Li ions, 64 MeV16O ions, and 80 MeV32S ions. The experimental radiation effectiveness (RE) with reference to low-LE...

Investigation of light-induced conformation changes in spiropyran-modified succinylated poly(L-lysine).

Science.gov (United States)

Cooper, T M; Stone, M O; Natarajan, L V; Crane, R L

1995-08-01

To determine the maximum range of coupling between side-chain photochromism and polypeptide conformation change, we modified the carboxylate side chains of succinylated poly(L-lysine) with a spiropyran to form polypeptide I. The extent of modification was determined to be 35.5%. The spacer group length between the polypeptide alpha-carbon and the dye was 12 atoms, providing minimum polypeptide-dye interaction. Conformation changes were monitored by circular dichroism as a function of light adaptation and solvent composition (hexafluoroisopropanol [HFIP] vs trifluoroethanol [TFE]). Under all solvent compositions, the dark-adapted dye was in the merocyanine form. Light adaptation by visible light converted the dye to the spiropyran form. When dissolved in TFE, I adopted a helical conformation insensitive to light adaptation. With increasing percentage HFIP, a solvent-induced helix-to-coil transition was observed around 80% (vol/vol) HFIP. At 100% HFIP, both light- and dark-adapted forms of I were in the coil state. Near the midpoint of the solvent-induced helix-to-coil transition, light adaptation caused conformation changes. Applying helix-to-coil transition theory, we measured a statistically significant difference in coil segment-HFIP binding constant for light- vs dark-adapted solutions (6.38 +/- 0.03 M-1 vs 6.56 +/- 0.03 M-1), but not for the nucleation parameter sigma (1.2 +/- 0.4 10(-3) vs 1.3 +/- 0.3 x 10(-3). The small binding constant difference translated to a light-induced binding energy difference of 17 cal/mol/monomer. Near the midpoint of the helix-to-coil transition, collective interactions between monomer units made possible the translation of a small energy difference (less than RT) into large macromolecular conformation changes.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of alpha interferon on glucose and alanine transport by rat renal brush border membrane vesicles

International Nuclear Information System (INIS)

Batuman, V.; Chadha, I.

1990-01-01

To investigate the pathogenetic mechanisms of interferon nephrotoxicity, we studied the effect of recombinant interferon alfa-2b on the uptake of 14 C-D-glucose and 14 C-L-alanine by rat renal brush-border-membrane vesicles. Interferon significantly inhibited 20 sec. sodium-dependent and 5 and 10 min. equilibrium uptake of both glucose and alanine. The inhibitory effect was dose dependent with maximum effect achieved at interferon concentration of 5 x 10 -8 M in the uptake media. The half-maximal inhibitory concentrations, IC 50 , of interferon on glucose uptake was 1.8 x 10 -8 M, and 5.4 x 10 -9 M on alanine uptake. Dixon plot analysis of uptake data was consistent with pure non-competitive inhibition. The inhibition constants, K i , 1.5 x 10 -8 M for glucose uptake, and 7.3 x 10 -9 M for alanine uptake, derived from Dixon plots were in close agreement with the IC 50 s calculated from the semilog dose response curves. These observations reveal that direct interactions at the proximal tubule cell membrane are involved in the pathogenesis of interferon nephrotoxicity, and that its mechanism of nephrotoxicity is similar to that of other low molecular weight proteins

Triple Helix going abroad?

DEFF Research Database (Denmark)

Sørensen, Olav Jull; Hu, Yimei

2014-01-01

The aim of the article is to explore to what extent the Tripple helix is being internationalized. Each of the helixes have their own internationalization rationale but the article show by small example that the helix itself is being internationalized and integrated with the host country tripple h...

Thermal degradation process of poly (alpha-methylstyrene) microspheres coated with glow discharge polymer

International Nuclear Information System (INIS)

Zhang Zhanwen; Huang Yong; Tang Yongjian; Li Bo; Chen Sufen; He Zhibing

2009-01-01

Glow discharge polymer (GDP) shell was made by the decomposable mandrel technique using poly(alpha-methylstyrene) (PAMS) mandrel. The PAMS degradation rate and the GDP shell surface morphology at different equilibrium temperatures were investigated. Degradation rate was calculated from weight variation of PAMS before and after pyrolysis process. Experiment results indicate that the degradation rate decreases at the fixed equilibrium temperature and graded temperature can improve the rate. The degradation process has an effect on the GDP shell properties. The PAMS doesn't molten to flow liquid during degradation. But the degradation can reduce surface finish of GDP coatings. The GDP shell deffects are the result of the PAMS degradiation process. (authors)

Salt- and pH-Triggered Helix-Coil Transition of Ionic Polypeptides under Physiology Conditions.

Science.gov (United States)

Yuan, Jingsong; Zhang, Yi; Sun, Yue; Cai, Zhicheng; Yang, Lijiang; Lu, Hua

2018-06-11

Controlling the helix-coil transition of polypeptides under physiological conditions is an attractive way toward smart functional materials. Here, we report the synthesis of a series of tertiary amine-functionalized ethylene glycol (EG x )-linked polypeptide electrolytes with their secondary structures tunable under physiological conditions. The resultant polymers, denoted as P(EG x DMA-Glu) ( x = 1, 2, and 3), show excellent aqueous solubility (>20 mg/mL) regardless of their charge states. Unlike poly-l-lysine that can form a helix only at pH above 10, P(EG x DMA-Glu) undergo a pH-dependent helix-coil switch with their transition points within the physiological range (pH ∼5.3-6.5). Meanwhile, P(EG x DMA-Glu) exhibit an unusual salt-induced helical conformation presumably owing to the unique properties of EG x linkers. Together, the current work highlights the importance of fine-tuning the linker chemistry in achieving conformation-switchable polypeptides and represents a facile approach toward stimuli-responsive biopolymers for advanced biological applications.

Characterizing the Secondary Protein Structure of Black Widow Dragline Silk Using Solid-State NMR & X-ray Diffraction

Science.gov (United States)

Jenkins, Janelle E.; Sampath, Sujatha; Butler, Emily; Kim, Jihyun; Henning, Robert W.; Holland, Gregory P.; Yarger, Jeffery L.

2013-01-01

This study provides a detailed secondary structural characterization of major ampullate dragline silk from Latrodectus hesperus (black widow) spiders. X-ray diffraction results show that the structure of black widow major ampullate silk fibers is comprised of stacked β-sheet nanocrystallites oriented parallel to the fiber axis and an amorphous region with oriented (anisotropic) and isotropic components. The combination of two-dimensional (2D) 13C-13C through-space and through-bond solid-state NMR experiments provide chemical shifts that are used to determine detailed information about amino acid motif secondary structure in black widow spider dragline silk. Individual amino acids are incorporated into different repetitive motifs that make up the majority of this protein-based biopolymer. From the solid-state NMR measurements, we assign distinct secondary conformations to each repetitive amino acid motif and hence to the amino acids that make up the motifs. Specifically, alanine is incorporated in β-sheet (poly(Alan) and poly(Gly-Ala)), 31-helix (poly(Gly-Gly-Xaa), and α-helix (poly(Gln-Gln-Ala-Tyr)) components. Glycine is determined to be in β-sheet (poly(Gly-Ala)) and 31-helical (poly(Gly-Gly-Xaa)) regions, while serine is present in β-sheet (poly(Gly-Ala-Ser)), 31-helix (poly(Gly-Gly-Ser)), and β-turn (poly(Gly-Pro-Ser)) structures. These various motif-specific secondary structural elements are quantitatively correlated to the primary amino acid sequence of major ampullate spidroin 1 and 2 (MaSp1 and MaSp2) and are shown to form a self-consistent model for black widow dragline silk. PMID:24024617

Living Polymerization of N -Substituted β-Alanine N -Carboxyanhydrides: Kinetic Investigations and Preparation of an Amphiphilic Block Copoly-β-Peptoid

KAUST Repository

Grossmann, Arlett; Luxenhofer, Robert

2012-01-01

attention. Here we present results that suggest that while N-substituted β-alanine N-carboxyanhydrides can undergo a living nucleophilic ring-opening polymerization, the solubility of poly(β-peptoid)s can be very poor, which contributes to the limited

The Effect of a Helix-Coil Transition on the Extension Elasticity

Science.gov (United States)

Buhot, Arnaud; Halperin, Avi

2000-03-01

The secondary structure of a polymer affects its deformation behavior in accordance with the Le Chatelier principle. An important example of such secondary structure is the alpha helix encountered in polypeptides. Similar structure was recently proposed for PEO in aqueous media. Our discussion concerns the coupling of the cooperative helix-coil transition and the extension elasticity. In particular, we analyze the extension of a long single chain by use of optical tweezers or AFM. We consider chains that exist in the coil-state when unperturbed. The transition nevertheless occurs because the extension favors the low entropy helical state. As a result, the corresponding force law exhibits a plateau. The analysis of this situation involves two ingredients: (I) the stretching free energy penalty for a rod-coil mutiblock copolymer (II) the entropy associated with the possible placements of the rod and coil blocks.

Adsorption of {alpha}-amylase onto poly(N-vinyl 2-pyrrolidone/itaconic acid) hydrogels

Energy Technology Data Exchange (ETDEWEB)

Tuemtuerk, Hayrettin; Caykara, Tuncer; Kantoglu, Oemer; Gueven, Olgun

1999-05-02

{alpha}-Amylase enzyme was adsorbed on poly(N-vinyl 2-pyrrolidone/itaconic acid) (P(VP/IA)) hydrogels prepared by irradiating the ternary mixtures of VP/IA/water by {gamma}-rays at ambient temperature. The adsorption capacity of the hydrogels was determined to increase from 2.30 to 3.40 mg {alpha}-amylase/g dry gel with increasing amount of IA in gel system. Kinetic parameters were calculated as 2.51 g/dm{sup 3} for K{sub m} and 1.67x10{sup -3} g/dm{sup 3} min for V{sub max} for free enzyme and in the range of 3.88-5.02 g/dm{sup 3} for K{sub m} and 1.62x10{sup -3}-2.27 x 10{sup -3} g/dm{sup 3} min for V{sub max} depending on the amount of IA in the hydrogel. Enzyme activities were found to increase from 49.9% to 77.4% with increasing amount of IA in the gel system and retained their activities for one month storage. On the other hand, the free enzyme loses its activity completely after 20 days.

Characterization of lipoteichoic acid structures from three probiotic Bacillus strains: involvement of D-alanine in their biological activity.

Science.gov (United States)

Villéger, Romain; Saad, Naima; Grenier, Karine; Falourd, Xavier; Foucat, Loïc; Urdaci, Maria C; Bressollier, Philippe; Ouk, Tan-Sothea

2014-10-01

Probiotics represent a potential strategy to influence the host's immune system thereby modulating immune response. Lipoteichoic Acid (LTA) is a major immune-stimulating component of Gram-positive cell envelopes. This amphiphilic polymer, anchored in the cytoplasmic membrane by means of its glycolipid component, typically consists of a poly (glycerol-phosphate) chain with D-alanine and/or glycosyl substitutions. LTA is known to stimulate macrophages in vitro, leading to secretion of inflammatory mediators such as Nitric Oxide (NO). This study investigates the structure-activity relationship of purified LTA from three probiotic Bacillus strains (Bacillus cereus CH, Bacillus subtilis CU1 and Bacillus clausii O/C). LTAs were extracted from bacterial cultures and purified. Chemical modification by means of hydrolysis at pH 8.5 was performed to remove D-alanine. The molecular structure of native and modified LTAs was determined by (1)H NMR and GC-MS, and their inflammatory potential investigated by measuring NO production by RAW 264.7 macrophages. Structural analysis revealed several differences between the newly characterized LTAs, mainly relating to their D-alanylation rates and poly (glycerol-phosphate) chain length. We observed induction of NO production by LTAs from B. subtilis and B. clausii, whereas weaker NO production was observed with B. cereus. LTA dealanylation abrogated NO production independently of the glycolipid component, suggesting that immunomodulatory potential depends on D-alanine substitutions. D-alanine may control the spatial configuration of LTAs and their recognition by cell receptors. Knowledge of molecular mechanisms behind the immunomodulatory abilities of probiotics is essential to optimize their use.

Alanine dosimetry for clinical applications. Proceedings

International Nuclear Information System (INIS)

Anton, M.

2006-05-01

The following topics are dealt with: Therapy level alanine dosimetry at the UK Nationational Physical Laboratory, alanine as a precision validation tool for reference dosimetry, composition of alanine pellet dosimeters, the angular dependence of the alanine ESR spectrum, the CIAE alanine dosimeter for radiotherapy level, a correction for temporal evolution effects in alanine dosimetry, next-generation services foe e-traceability to ionization radiation national standards, establishing e-traceability to HIST high-dose measurement standards, alanine dosimetry of dose delivery from clinical accelerators, the e-scan alanine dosimeter reader, alanine dosimetry at ISS, verification of the integral delivered dose for IMRT treatment in the head and neck region with ESR/alanine dosimetry, alanine dosimetry in helical tomotherapy beams, ESR dosimetry research and development at the University of Palermo, lithium formate as a low-dose EPR radiation dosimeter, sensitivity enhancement of alanine/EPR dosimetry. (HSI)

Synthesis and characterization of new nanocomposites films using alanine-Cu-functionalized graphene oxide as nanofiller and PVA as polymeric matrix for improving of their properties

Science.gov (United States)

Abdolmaleki, Amir; Mallakpour, Shadpour; Karshenas, Azam

2017-09-01

In the synthesis of polymer-graphene nanocomposites, for improving properties of nanocomposites, two factors dispersion and strong interfacial interactions between graphene and the polymer, are essential. In the present work, poly(vinyl alcohol) PVA/GO-Cu-alanine nanocomposite films were manufactured using concentrations 0, 1, 3 and 5 wt% of GO-Cu-alanine in water solution. For this purpose, L-alanine amino acid was located on the surface and edges of GO through copper(II) ion as a coordinating function. Then, flexible PVA/GO-Cu-alanine nanocomposite films were fabricated using GO-Cu-alanine as filler and PVA as matrix. Due to the existence of affective interaction between GO-Cu-alanine and PVA matrix, the acquired PVA/GO-Cu-alanine nanocomposites demonstrated great thermal and mechanical properties. Properties of manufactured materials were characterized by Fourier transform infrared, X-ray photoelectron spectroscopies (XPS), X-ray diffraction (XRD), Thermal gravimetric analysis, elemental analysis, field emission scanning electron microscopy, transmission electron microscopy and energy dispersive X-ray spectroscopy (EDX).

Triple helix-forming oligonucleotide corresponding to the polypyrimidine sequence in the rat alpha 1(I) collagen promoter specifically inhibits factor binding and transcription.

Science.gov (United States)

Kovacs, A; Kandala, J C; Weber, K T; Guntaka, R V

1996-01-19

Type I and III fibrillar collagens are the major structural proteins of the extracellular matrix found in various organs including the myocardium. Abnormal and progressive accumulation of fibrillar type I collagen in the interstitial spaces compromises organ function and therefore, the study of transcriptional regulation of this gene and specific targeting of its expression is of major interest. Transient transfection of adult cardiac fibroblasts indicate that the polypurine-polypyrimidine sequence of alpha 1(I) collagen promoter between nucleotides - 200 and -140 represents an overall positive regulatory element. DNase I footprinting and electrophoretic mobility shift assays suggest that multiple factors bind to different elements of this promoter region. We further demonstrate that the unique polypyrimidine sequence between -172 and -138 of the promoter represents a suitable target for a single-stranded polypurine oligonucleotide (TFO) to form a triple helix DNA structure. Modified electrophoretic mobility shift assays show that this TFO specifically inhibits the protein-DNA interaction within the target region. In vitro transcription assays and transient transfection experiments demonstrate that the transcriptional activity of the promoter is inhibited by this oligonucleotide. We propose that TFOs represent a therapeutic potential to specifically influence the expression of alpha 1(I) collagen gene in various disease states where abnormal type I collagen accumulation is known to occur.

beta-Chloro-L-alanine inhibition of the Escherichia coli alanine-valine transaminase.

OpenAIRE

Whalen, W A; Wang, M D; Berg, C M

1985-01-01

beta-Chloro-L-alanine, an amino acid analog which inhibits a number of enzymes, reversibly inhibited the Escherichia coli K-12 alanine-valine transaminase, transaminase C. This inhibition, along with the inhibition of transaminase B, accounted for the isoleucine-plus-valine requirement of E. coli in the presence of beta-chloro-L-alanine.

Capillary electrochromatography immunoassay for alpha-fetoprotein based on poly(guanidinium ionic liquid) monolithic material.

Science.gov (United States)

Liu, Cuicui; Deng, Qiliang; Fang, Guozhen; Dang, Meng; Wang, Shuo

2017-08-01

Alpha-fetoprotein (AFP) is widely used as a tumor marker for the serum diagnosis of primary hepatoma. Sensitive detection of AFP level plays an important role in the early diagnosis of disease and highly reliable prediction. In this study, a novel non-competitive immunoassay (IA) based on poly(guanidinium ionic liquid) monolithic material was developed for detecting ultra trace levels of AFP in capillary electrochromatography (CEC) mode. The AFP was mixed with an excess amount of fluorescently labeled antibody. After incubation, the immunocomplex was separated from the free labeled antibody and detected by CEC coupled with laser-induced fluorescence detector. Under the optimized conditions, the developed CEC-IA performed a low detection limit of 0.05 μg L -1  (S/N = 3) and a wide linearity ranging from 0.1 to 1000 μg L -1 for AFP, which can be largely attributed to the high separation and enrichment efficiency of poly(guanidinium ionic liquid) monolithic material for the targets. The application of this method was demonstrated by determining AFP in human serum. Copyright © 2017. Published by Elsevier Inc.

Conservation of a helix-stabilizing dipole moment in the PP-fold family of regulatory peptides

DEFF Research Database (Denmark)

Bjørnholm, B; Jørgensen, Flemming Steen; Schwartz, T W

1993-01-01

arrangement were performed in two ways: (1) by the use of a Poisson-Boltzmann approach which allows for an estimate of the screening effect, and (2) by the use of a uniform dielectric model (Coulomb's law). It is found that the alpha-helix is stabilized by approximately 5-10 kcal/mol due to electrostatic...

Production of D-alanine from DL-alanine using immobilized cells of Bacillus subtilis HLZ-68.

Science.gov (United States)

Zhang, Yangyang; Li, Xiangping; Zhang, Caifei; Yu, Xiaodong; Huang, Fei; Huang, Shihai; Li, Lianwei; Liu, Shiyu

2017-09-13

Immobilized cells of Bacillus subtilis HLZ-68 were used to produce D-alanine from DL-alanine by asymmetric degradation. Different compounds such as polyvinyl alcohol and calcium alginate were employed for immobilizing the B. subtilis HLZ-68 cells, and the results showed that cells immobilized using a mixture of these two compounds presented higher L-alanine degradation activity, when compared with free cells. Subsequently, the effects of different concentrations of polyvinyl alcohol and calcium alginate on L-alanine consumption were examined. Maximum L-alanine degradation was exhibited by cells immobilized with 8% (w/v) polyvinyl alcohol and 2% (w/v) calcium alginate. Addition of 400 g of DL-alanine (200 g at the beginning of the reaction and 200 g after 30 h of incubation) into the reaction solution at 30 °C, pH 6.0, aeration of 1.0 vvm, and agitation of 400 rpm resulted in complete L-alanine degradation within 60 h, leaving 185 g of D-alanine in the reaction solution. The immobilized cells were applied for more than 15 cycles of degradation and a maximum utilization rate was achieved at the third cycle. D-alanine was easily extracted from the reaction solution using cation-exchange resin, and the chemical and optical purity of the extracted D-alanine was 99.1 and 99.6%, respectively.

A study of the irradiation temperature coefficient for L-alanine and DL-alanine dosemeters

International Nuclear Information System (INIS)

Desrosiers, M. F.; Lin, M.; Cooper, S. L.; Cui, Y.; Chen, K.

2006-01-01

Alanine dosimetry is now well established both as a reference and routine dosemeter for industrial irradiation processing. Accurate dosimetry under the relatively harsh conditions of industrial processing requires a characterisation of the parameters that influence the dosemeter response. The temperature of the dosemeter during irradiation is a difficult quantity to measure so that the accuracy of the temperature coefficient that governs the dosemeter response becomes a critical factor. Numerous publications have reported temperature coefficients for several types of alanine dosemeters. The observed differences in the measured values were commonly attributed to the differences in the polymer binder or the experimental design of the measurement. However, the data demonstrated a consistent difference in the temperature coefficients between L-alanine and DL-alanine. Since there were no commonalities in the dosemeter composition or the measurement methods applied, a clear conclusion is not possible. To resolve this issue, the two isomeric forms of alanine dosemeters were prepared and irradiated in an identical manner. The results indicated that the DL-alanine temperature coefficient is more than 50% higher than the L-alanine temperature coefficient. (authors)

DNA binding specificity of the basic-helix-loop-helix protein MASH-1.

Science.gov (United States)

Meierhan, D; el-Ariss, C; Neuenschwander, M; Sieber, M; Stackhouse, J F; Allemann, R K

1995-09-05

Despite the high degree of sequence similarity in their basic-helix-loop-helix (BHLH) domains, MASH-1 and MyoD are involved in different biological processes. In order to define possible differences between the DNA binding specificities of these two proteins, we investigated the DNA binding properties of MASH-1 by circular dichroism spectroscopy and by electrophoretic mobility shift assays (EMSA). Upon binding to DNA, the BHLH domain of MASH-1 underwent a conformational change from a mainly unfolded to a largely alpha-helical form, and surprisingly, this change was independent of the specific DNA sequence. The same conformational transition could be induced by the addition of 20% 2,2,2-trifluoroethanol. The apparent dissociation constants (KD) of the complexes of full-length MASH-1 with various oligonucleotides were determined from half-saturation points in EMSAs. MASH-1 bound as a dimer to DNA sequences containing an E-box with high affinity KD = 1.4-4.1 x 10(-14) M2). However, the specificity of DNA binding was low. The dissociation constant for the complex between MASH-1 and the highest affinity E-box sequence (KD = 1.4 x 10(-14) M2) was only a factor of 10 smaller than for completely unrelated DNA sequences (KD = approximately 1 x 10(-13) M2). The DNA binding specificity of MASH-1 was not significantly increased by the formation of an heterodimer with the ubiquitous E12 protein. MASH-1 and MyoD displayed similar binding site preferences, suggesting that their different target gene specificities cannot be explained solely by differential DNA binding. An explanation for these findings is provided on the basis of the known crystal structure of the BHLH domain of MyoD.

Secretion of d-alanine by Escherichia coli.

Science.gov (United States)

Katsube, Satoshi; Sato, Kazuki; Ando, Tasuke; Isogai, Emiko; Yoneyama, Hiroshi

2016-07-01

Escherichia coli has an l-alanine export system that protects the cells from toxic accumulation of intracellular l-alanine in the presence of l-alanyl-l-alanine (l-Ala-l-Ala). When a DadA-deficient strain was incubated with 6.0 mM l-Ala-l-Ala, we detected l-alanine and d-alanine using high-performance liquid chromatography (HPLC) analysis at a level of 7.0 mM and 3.0 mM, respectively, after 48 h incubation. Treatment of the culture supernatant with d-amino acid oxidase resulted in the disappearance of a signal corresponding to d-alanine. Additionally, the culture supernatant enabled a d-alanine auxotroph to grow without d-alanine supplementation, confirming that the signal detected by HPLC was authentic d-alanine. Upon introduction of an expression vector harbouring the alanine racemase genes, alr or dadX, the extracellular level of d-alanine increased to 11.5 mM and 8.5 mM, respectively, under similar conditions, suggesting that increased metabolic flow from l-alanine to d-alanine enhanced d-alanine secretion. When high-density DadA-deficient cells preloaded with l-Ala-l-Ala were treated with 20 µM carbonyl cyanide m-chlorophenyl hydrazone (CCCP), secretion of both l-alanine and d-alanine was enhanced ~twofold compared with that in cells without CCCP treatment. In contrast, the ATPase inhibitor dicyclohexylcarbodiimide did not exert such an effect on the l-alanine and d-alanine secretion. Furthermore, inverted membrane vesicles prepared from DadA-deficient cells lacking the l-alanine exporter AlaE accumulated [3H]D-alanine in an energy-dependent manner. This energy-dependent accumulation of [3H]D-alanine was strongly inhibited by CCCP. These results indicate that E. coli has a transport system(s) that exports d-alanine and that this function is most likely modulated by proton electrochemical potential.

Comparison of alanine dosimeters using silicone as their binder to a commercial, polystyrene-bound, alanine dosimeter

International Nuclear Information System (INIS)

Galindo, S.; Urena-Nunez, F.

1997-01-01

The feasibility of practical boron-containing alanine ESR dosimeters for gamma-neutron mixed field irradiation dosimeters depends in part on whether the γ response characteristics of these silicone-bound dosimeters are comparable to those of a commercially available dosimeter that has been used by the International Atomic Energy Agency (International Dose Assurance Service) as a transfer reference dosimeter. This work presents the results of the comparison of 3 batches of silicone-bound alanine dosimeters. The first batch consists of a mixture of alanine and boric acid; the second, alanine and borax; and the last contains only alanine. Results indicate that γ response characteristics of the silicone-bound samples are comparable to those of the commercial, polystyrene-bound, alanine dosimeter and that silicone has a strong potential as a binding substance for alanine ESR dosimetry. (Author)

Defining the transmembrane helix of M2 protein from influenza A by molecular dynamics simulations in a lipid bilayer

NARCIS (Netherlands)

Forrest, LR; Tieleman, DP; Sansom, MSP

Integral membrane proteins containing at least one transmembrane (TM) alpha-helix are believed to account for between 20% and 30% of most genomes. There are several algorithms that accurately predict the number and position of TM helices within,a membrane protein sequence. However, these methods

Monopeptide versus Monopeptoid: Insights on Structure and Hydration of Aqueous Alanine and Sarcosine via X-ray Absorption Spectroscopy

Energy Technology Data Exchange (ETDEWEB)

Uejio, Janel S.; Schwartz, Craig P.; Duffin, Andrew M.; England, Alice; Prendergast, David; Saykally, Richard J.

2009-11-19

Despite the obvious significance, the aqueous interactions of peptides remain incompletely understood. Their synthetic analogues called peptoids (poly-N-substituted glycines), have recently emerged as a promising biomimetic material, particularly due to their robust secondary structure and resistance to denaturation. We describe comparative near-edge x-ray absorption fine structure (NEXAFS) spectroscopy studies of aqueous sarcosine, the simplest peptoid, and alanine, its peptide isomer, interpreted by density functional theory calculations. The sarcosine nitrogen K-edge spectrum is blue-shifted with respect to that of alanine, in agreement with our calculations; we conclude that this shift results primarily from the methyl group substitution on the nitrogen of sarcosine. Our calculations indicate that the nitrogen K-edge spectrum of alanine differs significantly between dehydrated and hydrated scenarios, while that of the sarcosine zwitterion is less affected by hydration. In contrast, the computed sarcosine spectrum is greatly impacted by conformational variations, while the alanine spectrum is not. This relates to a predicted solvent dependence for alanine, as compared to sarcosine. Additionally, we show the theoretical nitrogen K-edge spectra to be sensitive to the degree of hydration, indicating that experimental X-ray spectroscopy may be able to distinguish between bulk and partial hydration, such as found in confined environments near proteins and in reverse micelles.

Multiple helix ecosystems for sustainable competitiveness

CERN Document Server

Ferreira, João; Farinha, Luís; Fernandes, Nuno

2016-01-01

This book discusses the main issues, challenges, opportunities, and trends involving the interactions between academia, industry, government and society. Specifically, it aims to explore how these interactions enhance the ways in which companies deliver products and services in order to achieve sustainable competitiveness in the marketplace. Sustainable competitiveness has been widely discussed by academics and practitioners, considering the importance of protecting the environment while sustaining the economic goals of organizations. The Quintuple Helix innovation model is a framework for facilitating knowledge, innovation and sustainable competitive advantage. It embeds the Triple and the Quadruple Helix models by adding a fifth helix, the “natural environment.” The Triple Helix model focuses on the university-industry-government triad, while the Quadruple adds civil society (the media- and culture-driven public) as a fourth helix. The Quintuple Helix model facilitates research, public policy, and pract...

Effects of alpha radiation on plutonium incorporated in dosimetric materials by ESR studies

International Nuclear Information System (INIS)

Bhide, M.K.; Kadam, R.M.; Mohapatra, Manoj; Godbole, S.V.

2007-01-01

The in situ alpha irradiation effects of some ESR dosimetric materials namely alanine, 2-methyl alanine and ammonium tartrate were studied by incorporating 1% plutonium by weight in them. The radical intensity was monitored as a function of alpha dose. It was found that in the dose region 1-35 kGy ammonium tartrate showed better signal intensity, linearity and dose response as compared to the other materials. This was attributed to the single radical produced in case of the tartrate giving a sharp spectrum and the fast relaxation times owing to less saturation of ESR signals. (author)

Selection of D-Alanine-Tolerant Rice Cells

OpenAIRE

Hisashi, Manabe; Koji, Ohira; Aizu Junior College of Fukushima Prefecture; Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University

1984-01-01

By repeating subculture of rice cells (parent cells) in a D-alanine containing medium, we could select rice cells which grew well in the D-alanine medium. The D-alanine-tolerant cells absorbed a fairly small amount of D-alanine from the medium and did not accumulate much D-alanine in the cells. Aggregation of D-alanine-tolerant cells was greater than that of parent cells. D-Alanine metabolism of D-alanine.-tolerant cells did not increase in comparison with parent cells.

Comparison of EPR response of alanine and Gd₂O₃-alanine dosimeters exposed to TRIGA Mainz reactor.

Science.gov (United States)

Marrale, M; Schmitz, T; Gallo, S; Hampel, G; Longo, A; Panzeca, S; Tranchina, L

2015-12-01

In this work we report some preliminary results regarding the analysis of electron paramagnetic resonance (EPR) response of alanine pellets and alanine pellets added with gadolinium used for dosimetry at the TRIGA research reactor in Mainz, Germany. Two set-ups were evaluated: irradiation inside PMMA phantom and irradiation inside boric acid phantom. We observed that the presence of Gd2O3 inside alanine pellets increases the EPR signal by a factor of 3.45 and 1.24 in case of PMMA and boric acid phantoms, respectively. We can conclude that in the case of neutron beam with a predominant thermal neutron component the addition of gadolinium oxide can significantly improve neutron sensitivity of alanine pellets. Monte Carlo (MC) simulations of both response of alanine and Gd-added alanine pellets with FLUKA code were performed and a good agreement was achieved for pure alanine dosimeters. For Gd2O3-alanine deviations between MC simulations and experimental data were observed and discussed. Copyright © 2015 Elsevier Ltd. All rights reserved.

Effect of TFE on the Helical Content of AK17 and HAL-1 Peptides: Theoretical Insights into the Mechanism of Helix Stabilization.

Science.gov (United States)

Vymětal, Jiří; Bednárová, Lucie; Vondrášek, Jiří

2016-02-18

Fluorinated alcohols such as 2,2,2-trifluoroethanol (TFE) are among the most frequently used cosolvents in experiment studies of peptides. They have significant effects on secondary structure and a particularly strong promotion of α-helix is induced by TFE. In this study we validated recently proposed force field parameters for TFE in molecular dynamics simulations with two model peptides-alanine-rich AK-17 and antimicrobial peptide halictine-1 (HAL-1). In the case of HAL-1, we characterized the effect of TFE on this peptide experimentally by ECD spectroscopy. Our TFE model in question reproduced the helix-promoting effect of TFE and provided insight into the mechanisms of TFE action on peptides. Our simulations confirmed the preferential interaction of TFE molecules with α-helices, although the TFE molecules accumulate in the vicinity of the peptides in various conformations. Moreover, we observed a significant effect of TFE on the thermodynamics of the helix-coil transition and a change in local conformational preferences in the unfolded (coil) state induced by TFE. In addition, our simulation-based analysis suggests that different mechanisms participate in helix stabilization in both model peptides in water and TFE solution. Our results thus support the picture of complex TFE action on peptides that is further diversified by the identity and intrinsic properties of the peptide.

Effect of Exogenous _D-Alanine on _D-Alanyl-_D-alanine Content in Leaf Blades of Oryza australiensis Domin

OpenAIRE

Hisashi, Manabe; Aizu Junior College of Fukushima Prefecture

1986-01-01

In seedlings of Oryza australiensis Domin (W0008), most of the D-alanyl-D-alanine was distributed in the leaf blades. In excised leaf blades of W0008, exogenous D-alanine was incorporated into D-alanyl-D-alanine irrespective of the light condition as in Sasanishiki. With cultivation in D-alanine medium for several days, the D-alanyl-D-alanine content in W0008 leaf blades was found to increase, but no other D-alanine-containing dipeptides such as D-alanylglycine or D-alanyl-L-alanine were dete...

Increased helix and protein stability through the introduction of a new tertiary hydrogen bond.

Science.gov (United States)

Peterson, R W; Nicholson, E M; Thapar, R; Klevit, R E; Scholtz, J M

1999-03-12

In an effort to quantify the importance of hydrogen bonding and alpha-helix formation to protein stability, a capping box motif was introduced into the small phosphocarrier protein HPr. Previous studies had confirmed that Ser46, at the N-cap position of the short helix-B in HPr, serves as an N-cap in solution. Thus, only a single-site mutation was required to produce a canonical S-X-X-E capping box: Lys49 at the N3 position was substituted with a glutamic acid residue. Thermal and chemical denaturation studies on the resulting K49E HPr show that the designed variant is approximately 2 kcal mol-1 more stable than the wild-type protein. However, NMR studies indicate that the side-chain of Glu49 does not participate in the expected capping H-bond interaction, but instead forms a new tertiary H-bond that links helix-B to the four-stranded beta-sheet of HPr. Here, we demonstrate that a strategy in which new non-native H-bonds are introduced can generate proteins with increased stability. We discuss why the original capping box design failed, and compare the energetic consequences of the new tertiary side-chain to main-chain H-bond with a local (helix-capping) side-chain to main-chain H-bond on the protein's global stability. Copyright 1999 Academic Press.

Global Transcriptional and Physiological Responses of Saccharomyces cerevisiae to Ammonium, L-Alanine, or L-Glutamine Limitation

DEFF Research Database (Denmark)

Usaite, Renata; Patil, Kiran Raosaheb; Grotkjær, Thomas

2006-01-01

-ammonium in limitation and by growing cells in an excess of ammonium. Cells grown in L-alanine-limited cultures had higher biomass yield per nitrogen mole (19%) than those from ammonium-limited cultures. Whole-genome transcript profiles were analyzed with a genome-scalle metabolic model that suggested increased anabolic...... activity in L-alanine-limited cells. The changes in these cells were found to be focused around pyruvate, acetyl coenzyme A, glyoxylate, and alpha-ketoglutarate via increased levels of ALT1, DAL7, PYC1, GDH2, and ADH5 and decreased levels of GDH3, CIT2, and ACS1 transcripts. The transcript profiles were...

Introduction of potential helix-capping residues into an engineered helical protein.

Science.gov (United States)

Parker, M H; Hefford, M A

1998-08-01

MB-1 is an engineered protein that was designed to incorporate high percentages of four amino acid residues and to fold into a four-alpha-helix bundle motif. Mutations were made in the putative loop I and III regions of this protein with the aim of increasing the stability of the helix ends. Four variants, MB-3, MB-5, MB-11 and MB-13, have replacements intended to promote formation of an 'N-capping box'. The loop I and III sequences of MB-3 (both GDLST) and MB-11 (GGDST) were designed to cause alphaL C-terminal 'capping' motifs to form in helices I and III. MB-5 has a sequence, GPDST, that places proline in a favourable position for forming beta-turns, whereas MB-13 (GLDST) has the potential to form Schellman C-capping motifs. Size-exclusion chromatography suggested that MB-1, MB-3, MB-5, MB-11 and MB-13 all form dimers, or possibly trimers. Free energies for the unfolding of each of these variants were determined by urea denaturation, with the loss of secondary structure followed by CD spectroscopy. Assuming an equilibrium between folded dimer and unfolded monomer, MB-13 had the highest apparent stability (40.5 kJ/mol, with +/-2.5 kJ/mol 95% confidence limits), followed by MB-11 (39.3+/-5.9 kJ/mol), MB-3 (36.4+/-1.7 kJ/mol), MB-5 (34.7+/-2.1 kJ/mol) and MB-1 (29.3+/-1.3 kJ/mol); the same relative stabilities of the variants were found when a folded trimer to unfolded monomer model was used to calculate stabilities. All of the variants were relatively unstable for dimeric proteins, but were significantly more stable than MB-1. These findings suggest that it might be possible to increase the stability of a protein for which the three-dimensional structure is unknown by placing amino acid residues in positions that have the potential to form helix- and turn-stabilizing motifs.

Generating structured light with phase helix and intensity helix using reflection-enhanced plasmonic metasurface at 2 μm

Science.gov (United States)

Zhao, Yifan; Du, Jing; Zhang, Jinrun; Shen, Li; Wang, Jian

2018-04-01

Mid-infrared (2-20 μm) light has been attracting great attention in many areas of science and technology. Beyond the extended wavelength range from visible and near-infrared to mid-infrared, shaping spatial structures may add opportunities to grooming applications of mid-infrared photonics. Here, we design and fabricate a reflection-enhanced plasmonic metasurface and demonstrate efficient generation of structured light with the phase helix and intensity helix at 2 μm. This work includes two distinct aspects. First, structured light (phase helix, intensity helix) generation at 2 μm, which is far beyond the ability of conventional spatial light modulators, is enabled by the metasurface with sub-wavelength engineered structures. Second, the self-referenced intensity helix against environmental noise is generated without using a spatially separated light. The demonstrations may open up advanced perspectives to structured light applications at 2 μm, such as phase helix for communications and non-communications (imaging, sensing) and intensity helix for enhanced microscopy and advanced metrology.

A Classification of Basic Helix-Loop-Helix Transcription Factors of Soybean

Directory of Open Access Journals (Sweden)

Karen A. Hudson

2015-01-01

Full Text Available The complete genome sequence of soybean allows an unprecedented opportunity for the discovery of the genes controlling important traits. In particular, the potential functions of regulatory genes are a priority for analysis. The basic helix-loop-helix (bHLH family of transcription factors is known to be involved in controlling a wide range of systems critical for crop adaptation and quality, including photosynthesis, light signalling, pigment biosynthesis, and seed pod development. Using a hidden Markov model search algorithm, 319 genes with basic helix-loop-helix transcription factor domains were identified within the soybean genome sequence. These were classified with respect to their predicted DNA binding potential, intron/exon structure, and the phylogeny of the bHLH domain. Evidence is presented that the vast majority (281 of these 319 soybean bHLH genes are expressed at the mRNA level. Of these soybean bHLH genes, 67% were found to exist in two or more homeologous copies. This dataset provides a framework for future studies on bHLH gene function in soybean. The challenge for future research remains to define functions for the bHLH factors encoded in the soybean genome, which may allow greater flexibility for genetic selection of growth and environmental adaptation in this widely grown crop.

21 CFR 172.540 - DL-Alanine.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 3 2010-04-01 2009-04-01 true DL-Alanine. 172.540 Section 172.540 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Agents and Related Substances § 172.540 DL-Alanine. DL-Alanine (a racemic mixture of D- and L-alanine...

21 CFR 582.5118 - Alanine.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Alanine. 582.5118 Section 582.5118 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL DRUGS, FEEDS... § 582.5118 Alanine. (a) Product. Alanine (L- and DL-forms). (b) Conditions of use. This substance is...

Composite poly-L-lactic acid/poly-({alpha},{beta})-DL-aspartic acid/collagen nanofibrous scaffolds for dermal tissue regeneration

Energy Technology Data Exchange (ETDEWEB)

Ravichandran, Rajeswari [Healthcare and Energy Materials Laboratory, Nanoscience and Nanotechnology Initiative, Faculty of Engineering, National University of Singapore, 117576 (Singapore); Department of Mechanical Engineering, National University of Singapore, 117576 (Singapore); Venugopal, Jayarama Reddy, E-mail: nnijrv@nus.edu.sg [Healthcare and Energy Materials Laboratory, Nanoscience and Nanotechnology Initiative, Faculty of Engineering, National University of Singapore, 117576 (Singapore); Sundarrajan, Subramanian [Healthcare and Energy Materials Laboratory, Nanoscience and Nanotechnology Initiative, Faculty of Engineering, National University of Singapore, 117576 (Singapore); Department of Mechanical Engineering, National University of Singapore, 117576 (Singapore); Mukherjee, Shayanti [Healthcare and Energy Materials Laboratory, Nanoscience and Nanotechnology Initiative, Faculty of Engineering, National University of Singapore, 117576 (Singapore); Sridhar, Radhakrishnan [Healthcare and Energy Materials Laboratory, Nanoscience and Nanotechnology Initiative, Faculty of Engineering, National University of Singapore, 117576 (Singapore); Department of Mechanical Engineering, National University of Singapore, 117576 (Singapore); Ramakrishna, Seeram, E-mail: seeram@nus.edu.sg [Healthcare and Energy Materials Laboratory, Nanoscience and Nanotechnology Initiative, Faculty of Engineering, National University of Singapore, 117576 (Singapore); Department of Mechanical Engineering, National University of Singapore, 117576 (Singapore)

2012-08-01

Tissue engineering scaffolds for skin tissue regeneration is an ever expounding area of research, as the products that meet the necessary requirements are far and elite. The nanofibrous poly-L-lactic acid/poly-({alpha},{beta})-DL-aspartic acid/Collagen (PLLA/PAA/Col I and III) scaffolds were fabricated by electrospinning and characterized by SEM, contact angle and FTIR analysis for skin tissue regeneration. The cell-scaffold interactions were analyzed by cell proliferation and their morphology observed in SEM. The results showed that the cell proliferation was significantly increased (p {<=} 0.05) in PLLA/PAA/Col I and III scaffolds compared to PLLA and PLLA/PAA nanofibrous scaffolds. The abundance and accessibility of adipose derived stem cells (ADSCs) may prove to be novel cell therapeutics for dermal tissue regeneration. The differentiation of ADSCs was confirmed using collagen expression and their morphology by CMFDA dye extrusion technique. The current study focuses on the application of PLLA/PAA/Col I and III nanofibrous scaffolds for skin tissue engineering and their potential use as substrate for the culture and differentiation of ADSCs. The objective for inclusion of a novel cell binding moiety like PAA was to replace damaged extracellular matrix and to guide new cells directly into the wound bed with enhanced proliferation and overall organization. This combinatorial epitome of PLLA/PAA/Col I and III nanofibrous scaffold with stem cell therapy to induce the necessary paracrine signalling effect would favour faster regeneration of the damaged skin tissues. - Highlights: Black-Right-Pointing-Pointer Differentiation of adipose derived stem cells in the presence of bFGF for wound healing Black-Right-Pointing-Pointer Introduction of PAA as ECM mimetic cell binding moiety Black-Right-Pointing-Pointer Combination of PLLA/PAA/Col I and III nanofibers and stem cell therapy for skin regeneration.

Oxidation of Helix-3 methionines precedes the formation of PK resistant PrP.

Directory of Open Access Journals (Sweden)

Tamar Canello

2010-07-01

Full Text Available While elucidating the peculiar epitope of the alpha-PrP mAb IPC2, we found that PrPSc exhibits the sulfoxidation of residue M213 as a covalent signature. Subsequent computational analysis predicted that the presence of sulfoxide groups at both Met residues 206 and 213 destabilize the alpha-fold, suggesting oxidation may facilitate the conversion of PrPC into PrPSc. To further study the effect of oxidation on prion formation, we generated pAbs to linear PrP peptides encompassing the Helix-3 region, as opposed to the non-linear complexed epitope of IPC2. We now show that pAbs, whose epitopes comprise Met residues, readily detected PrPC, but could not recognize most PrPSc bands unless they were vigorously reduced. Next, we showed that the alpha-Met pAbs did not recognize newly formed PrPSc, as is the case for the PK resistant PrP present in lines of prion infected cells. In addition, these reagents did not detect intermediate forms such as PK sensitive and partially aggregated PrPs present in infected brains. Finally, we show that PrP molecules harboring the pathogenic mutation E200K, which is linked to the most common form of familial CJD, may be spontaneously oxidized. We conclude that the oxidation of methionine residues in Helix-3 represents an early and important event in the conversion of PrPC to PrPSc. We believe that further investigation into the mechanism and role of PrP oxidation will be central in finally elucidating the mechanism by which a normal cell protein converts into a pathogenic entity that causes fatal brain degeneration.

Origin and Diversification of Basic-Helix-Loop-Helix Proteins in Plants

OpenAIRE

Pires, Nuno; Dolan, Liam

2009-01-01

Basic helix-loop-helix (bHLH) proteins are a class of transcription factors found throughout eukaryotic organisms. Classification of the complete sets of bHLH proteins in the sequenced genomes of Arabidopsis thaliana and Oryza sativa (rice) has defined the diversity of these proteins among flowering plants. However, the evolutionary relationships of different plant bHLH groups and the diversity of bHLH proteins in more ancestral groups of plants are currently unknown. In this study, we use wh...

Recognition and Binding of a Helix-Loop-Helix Peptide to Carbonic Anhydrase Occurs via Partly Folded Intermediate Structures

Science.gov (United States)

Lignell, Martin; Becker, Hans-Christian

2010-01-01

Abstract We have studied the association of a helix-loop-helix peptide scaffold carrying a benzenesulfonamide ligand to carbonic anhydrase using steady-state and time-resolved fluorescence spectroscopy. The helix-loop-helix peptide, developed for biosensing applications, is labeled with the fluorescent probe dansyl, which serves as a polarity-sensitive reporter of the binding event. Using maximum entropy analysis of the fluorescence lifetime of dansyl at 1:1 stoichiometry reveals three characteristic fluorescence lifetime groups, interpreted as differently interacting peptide/protein structures. We characterize these peptide/protein complexes as mostly bound but unfolded, bound and partly folded, and strongly bound and folded. Furthermore, analysis of the fluorescence anisotropy decay resulted in three different dansyl rotational correlation times, namely 0.18, 1.2, and 23 ns. Using the amplitudes of these times, we can correlate the lifetime groups with the corresponding fluorescence anisotropy component. The 23-ns rotational correlation time, which appears with the same amplitude as a 17-ns fluorescence lifetime, shows that the dansyl fluorophore follows the rotational diffusion of carbonic anhydrase when it is a part of the folded peptide/protein complex. A partly folded and partly hydrated interfacial structure is manifested in an 8-ns dansyl fluorescence lifetime and a 1.2-ns rotational correlation time. This structure, we believe, is similar to a molten-globule-like interfacial structure, which allows segmental movement and has a higher degree of solvent exposure of dansyl. Indirect excitation of dansyl on the helix-loop-helix peptide through Förster energy transfer from one or several tryptophans in the carbonic anhydrase shows that the helix-loop-helix scaffold binds to a tryptophan-rich domain of the carbonic anhydrase. We conclude that binding of the peptide to carbonic anhydrase involves a transition from a disordered to an ordered structure of the

An unusual helix-turn-helix protease inhibitory motif in a novel trypsin inhibitor from seeds of Veronica (Veronica hederifolia L.).

Science.gov (United States)

Conners, Rebecca; Konarev, Alexander V; Forsyth, Jane; Lovegrove, Alison; Marsh, Justin; Joseph-Horne, Timothy; Shewry, Peter; Brady, R Leo

2007-09-21

The storage tissues of many plants contain protease inhibitors that are believed to play an important role in defending the plant from invasion by pests and pathogens. These proteinaceous inhibitor molecules belong to a number of structurally distinct families. We describe here the isolation, purification, initial inhibitory properties, and three-dimensional structure of a novel trypsin inhibitor from seeds of Veronica hederifolia (VhTI). The VhTI peptide inhibits trypsin with a submicromolar apparent K(i) and is expected to be specific for trypsin-like serine proteases. VhTI differs dramatically in structure from all previously described families of trypsin inhibitors, consisting of a helix-turn-helix motif, with the two alpha helices tightly associated by two disulfide bonds. Unusually, the crystallized complex is in the form of a stabilized acyl-enzyme intermediate with the scissile bond of the VhTI inhibitor cleaved and the resulting N-terminal portion of the inhibitor remaining attached to the trypsin catalytic serine 195 by an ester bond. A synthetic, truncated version of the VhTI peptide has also been produced and co-crystallized with trypsin but, surprisingly, is seen to be uncleaved and consequently forms a noncovalent complex with trypsin. The VhTI peptide shows that effective enzyme inhibitors can be constructed from simple helical motifs and provides a new scaffold on which to base the design of novel serine protease inhibitors.

Chirality-specific lift forces of helix under shear flows: Helix perpendicular to shear plane.

Science.gov (United States)

Zhang, Qi-Yi

2017-02-01

Chiral objects in shear flow experience a chirality-specific lift force. Shear flows past helices in a low Reynolds number regime were studied using slender-body theory. The chirality-specific lift forces in the vorticity direction experienced by helices are dominated by a set of helix geometry parameters: helix radius, pitch length, number of turns, and helix phase angle. Its analytical formula is given. The chirality-specific forces are the physical reasons for the chiral separation of helices in shear flow. Our results are well supported by the latest experimental observations. © 2016 Wiley Periodicals, Inc.

Functional Characterization of Alanine Racemase from Schizosaccharomyces pombe: a Eucaryotic Counterpart to Bacterial Alanine Racemase

OpenAIRE

Uo, Takuma; Yoshimura, Tohru; Tanaka, Naotaka; Takegawa, Kaoru; Esaki, Nobuyoshi

2001-01-01

Schizosaccharomyces pombe has an open reading frame, which we named alr1+, encoding a putative protein similar to bacterial alanine racemase. We cloned the alr1+ gene in Escherichia coli and purified the gene product (Alr1p), with an Mr of 41,590, to homogeneity. Alr1p contains pyridoxal 5′-phosphate as a coenzyme and catalyzes the racemization of alanine with apparent Km and Vmax values as follows: for l-alanine, 5.0 mM and 670 μmol/min/mg, respectively, and for d-alanine, 2.4 mM and 350 μmo...

Synthesis, physicochemical and biological properties of poly-α-amino acids - the simplest of protein models

International Nuclear Information System (INIS)

Katchalski-Katzir, Ephraim

1996-01-01

During the 1950s, linear and multichain poly-α-amino acids were synthesized by polymerization of the corresponding N-carboxy-amino acid anhydrides in solution in the presence of suitable catalysts. The resulting homo- and heteropolymers have since been widely employed as simple protein models. Under appropriate conditions, poly-α-amino acids, in the solid state and in solution, were found to acquire conformations of an α-helix and β-parallel and antiparallel pleased sheets, or to exist as random coils. Their use in experimental and theoretical investigations of helix-coil transitions helped to shed new light on the mechanisms involved in protein denaturation. Poly-α-amino acids played an important role in the deciphering of the genetic code. In addition, analysis of the antigenicity of poly-α-amino acids led to the clucidation of the factors determining the antigenicity of proteins and peptides. Interest in the biological and physicochemical characteristics of poly-α-amino acids was recently renewed because of the reported novel finding that some copolymers of amino acids are effective as drugs in multiple sclerosis, and that glutamine repeats and reiteration of other amino acids occur in inherited neurodegenerative diseases. The presence of repeating sequences of amino acids in proteins, and of nucleotides in DNA, raises many interesting questions about their respective roles in determining protein structure and function, and gene performance and regulation. (author). 35 refs, 3 figs, 2 tabs

Highly Efficient Fumed Silica Nanoparticles for Peptide Bond Formation: Converting Alanine to Alanine Anhydride.

Science.gov (United States)

Guo, Chengchen; Jordan, Jacob S; Yarger, Jeffery L; Holland, Gregory P

2017-05-24

In this work, thermal condensation of alanine adsorbed on fumed silica nanoparticles is investigated using thermal analysis and multiple spectroscopic techniques, including infrared (IR), Raman, and nuclear magnetic resonance (NMR) spectroscopies. Thermal analysis shows that adsorbed alanine can undergo thermal condensation, forming peptide bonds within a short time period and at a lower temperature (∼170 °C) on fumed silica nanoparticle surfaces than that in bulk (∼210 °C). Spectroscopic results further show that alanine is converted to alanine anhydride with a yield of 98.8% during thermal condensation. After comparing peptide formation on solution-derived colloidal silica nanoparticles, it is found that fumed silica nanoparticles show much better efficiency and selectivity than solution-derived colloidal silica nanoparticles for synthesizing alanine anhydride. Furthermore, Raman spectroscopy provides evidence that the high efficiency for fumed silica nanoparticles is likely related to their unique surface features: the intrinsic high population of strained ring structures present at the surface. This work indicates the great potential of fumed silica nanoparticles in synthesizing peptides with high efficiency and selectivity.

Procedure for preparation of 3-fluor-D-alanine, 2-deutero-3-fluor-D-alanine and 2,3,3-trideutero-3-fluor-D-alanine and their salts

International Nuclear Information System (INIS)

Kollonitsch, J.; Kahan, F.M.

1971-01-01

Procedures for the preparation of 3-fluor-D-alanine, 2-deutero-3-fluor-D-alanine and 2,3,3-trideutero-3-fluor-D-alanine, and salts of these compounds, are described. These new compounds are useful antibacterial substances not only applicable in the disinfection of pharmaceutical, dental and medical equipment, but also in the treatment of diseases caused by bacteria, and may be administered orally. While 3-fluor-L-alanine metabolises rapidly with toxic results, 3-fluor-D-alanine is much more slowly broken down in vivo and is not harmful in normal doses. Further it has been found that deuteration gives new deutero-analogues which are less subject to metabolic breaking down and still retain the antibacterial strength of the original compound. The in vivo activity is thereby increased and maintained. (JIW)

Strain-induced large spin splitting and persistent spin helix at LaAlO$_3$/SrTiO$_3$ interface

OpenAIRE

Yamaguchi, Naoya; Ishii, Fumiyuki

2017-01-01

We investigated the effect of the tensile strain on the spin splitting at the n-type interface in LaAlO$_3$/SrTiO$_3$ in terms of the spin-orbit coupling coefficient $\\alpha$ and spin texture in the momentum space using first-principles calculations. We found that the $\\alpha$ could be controlled by the tensile strain and be enhanced up to 5 times for the tensile strain of 7%, and the effect of the tensile strain leads to a persistent spin helix, which has a long spin lifetime. These results ...

Physical data from CTD and bottle casts from the R/V ALPHA HELIX in the Gulf of Alaska by the University of Alaska; Institute of Marine Science (UAK/IMS) in support of the Exxon Oil Spill Monitoring Program and the Gulf of Alaska Recirculation Project from 1989-05-05 to 1989-05-28 (NODC Accession 8900171)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — Temperature, salinity, and sigma-t profiles from CTD and bottle casts from the R/V ALPHA HELIX. Data were collected in the Gulf of Alaska by the University of...

General synthesis of C-glycosyl amino acids via proline-catalyzed direct electrophilic alpha-amination of C-glycosylalkyl aldehydes.

Science.gov (United States)

Nuzzi, Andrea; Massi, Alessandro; Dondoni, Alessandro

2008-10-16

Non-natural axially and equatorially linked C-glycosyl alpha-amino acids (glycines, alanines, and CH2-serine isosteres) with either S or R alpha-configuration were prepared by D- and L-proline-catalyzed (de >95%) alpha-amination of C-glycosylalkyl aldehydes using dibenzyl azodicarboxylate as the electrophilic reagent.

Alanine flux in obese and healthy humans as evaluated by 15N- and 2H3-labeled alanines

International Nuclear Information System (INIS)

Hoffer, L.J.; Yang, R.D.; Matthews, D.E.; Bistrian, B.R.; Bier, D.M.; Young, V.R.

1988-01-01

Estimates of plasma alanine flux as measured in humans using L-[ 15 N]-alanine or L-[3,3,3- 2 H 3 ]alanine were compared by simultaneous intravenous infusion of both tracers. Plasma isotope enrichments were measured by chemical ionization gas chromatography-mass spectrometry. In 16 obese women before and during a hypocaloric diet and in 4 normal men in the postabsorptive and fed states, the fluxes were highly correlated (r2 = 0.93) although plasma alanine flux with the 2 H tracer was two to three times greater than that obtained with [ 15 N]alanine. The fluxes decreased with the hypocaloric diet in obese subjects and increased during the fed state in healthy adults. Thus, although the estimates of alanine flux differed according to the tracer used, both appear to give equivalent information about changes in alanine kinetics induced by the nutritional conditions examined

Helix-Hopes on Finite Hyperfields

Directory of Open Access Journals (Sweden)

Thomas Vougiouklis

2016-12-01

Full Text Available Hyperstructure theory can overcome restrictions which ordinary algebraic structures have. A hyperproduct on non-square ordinary matrices can be defined by using the so called helix-hyperoperations. We study the helix-hyperstructures on the representations using ordinary fields. The related theory can be faced by defining the hyperproduct on the set of non square matrices. The main tools of the Hyperstructure Theory are the fundamental relations which connect the largest class of hyperstructures, the Hv-structures, with the corresponding classical ones. We focus on finite dimensional helix-hyperstructures and on small Hv-fields, as well.

New optically active and thermally stable poly(amide-imide)s containing N,N'-(Bicyclo[2,2,2]oct-7-ene-2,3,5,6-tetracarboxylic)-bis-L-alanine and aromatic diamines: synthesis and characterization

International Nuclear Information System (INIS)

Faghihi, Khalil; Absalar, Morteza; Hajibeygi, Mohsen

2009-01-01

Five new optically active poly(amide-imide)s (PAIs) 6a-e were prepared by direct polycondensation reaction of the newly synthesized N,N'-(bicyclo[2,2,2]oct-7-ene-2,3,5,6-tetra carboxylic)-bis-L-alanine 4 with various aromatic diamines 5a-e using polar aprotic solvents such as N-methyl-2-pyrrolidone (NMP). In this technique triphenyl phosphite (TPP) and pyridine were used as condensing agents to form poly(amide-imide)s through the N-phosphonium salts of pyridine. All of the polymers were obtained in quantitative yields with inherent viscosities between 0.29-0.46 dL g -1 and were highly soluble in polar aprotic solvents such as N,N-dimethyl acetamide (DMAc), N,N-dimethyl formamide (DMF), dimethyl sulfoxide (DMSO), N-methyl- 2-pyrrolidone (NMP) and solvents such as sulfuric acid. They were fully characterized by means of 1 H NMR, FTIR spectroscopy, elemental analyses, inherent viscosity, solubility test, specific rotation and thermal properties of these polymers were investigated using thermogravimetric analysis techniques (TGA and DTG). (author)

Titration of poly(dA-dT) . poly(dA-dT) in solution at variable NaCl concentration.

Science.gov (United States)

Airoldi, Marta; Boicelli, C Andrea; Cadoni, Fabio; Gennaro, Giuseppe; Giomini, Marcello; Giuliani, Anna M; Giustini, Mauro

2004-10-05

CD and uv absorption data showed that high molecular weight poly(dA-dT) . poly(dA-dT), at 298 K, undergoes an acid-induced transition from B-double helix to random coil in NaCl solutions of different concentrations, ranging from 0.005 to 0.600M. Similarly, titration of the polynucleotide with a strong base causes duplex-to-single strands transition. The base- and acid-induced transitions were both reversible by back-titration (with an acid or, respectively, with a base): the apparent pKa were the same in both directions. However, the number of protons per titratable site (adenine N1) required to reach half-denaturation was in great excess over the stoichiometric value; to a much larger extent, the same effect was observed also for the deprotonation of the N3H sites of thymine. Moreover, in the basic denaturation experiments, at low salt concentrations ([NaCl]acid than calculated was needed to back-titrate the base excess to half-denaturation. Both effects could be qualitatively justified on the basis of the counterion condensation theory of polyelectrolytes and considering the energy barrier created by the negatively charged phosphodiester groups to the penetration of the OH- ions inside the double helix and the screening effect of the Na+ ions on such charges, in the deprotonation experiments.

Simultaneous determination of F-beta-alanine and beta-alanine in plasma and urine with dual-column reversed-phase high-performance liquid chromatography

NARCIS (Netherlands)

van Kuilenburg, A. B.; Stroomer, A. E.; Peters, G. J.; van Gennip, A. H.

2001-01-01

F-beta-Alanine and beta-alanine were detected in plasma and urine samples with fluorescence detection of orthophthaldialdehyde derivatives of F-beta-alanine and beta-alanine after separation with dual-column reversed-phase HPLC. The detection limits of F-beta-alanine and beta-alanine in the HPLC

Water-resistant alanine-EPR dosimeter alanpol

International Nuclear Information System (INIS)

Peimel-Stuglik, Zofia; Bryl-Sandelewska, Teresa; Mirkowski, Krzysztof; Sartowska, Bozena

2009-01-01

Alanpol-water-resistant alanine-electron paramagnetic resonance (EPR) dosimeter consisted of cheap DL-α-alanine (9.8-27%) suspended in polyethylene matrix was presented. The rods (O=2.8 mm) were extruded from a hot mixture of alanine and low-density polyethylene. No grinding or crushing was used for alanine preparation. An orientation of cylindrical crystals, up to 300 μm long in parallel to the rod axis was responsible for some differences in a shape of EPR signal. These differences had no negative consequences for dosimetric applications. Signal-to-dose dependence was linear up to 10 kGy. Standard deviation of dosimetric answer was up to ±1.8% and up to 2.4% for dosimeters with 9.8% and 27% of DL-α-alanine, respectively. Irradiation temperature coefficient for both dosimeters was equal 0.2%/ deg. C. Hydrophobic properties of polyethylene and small number of alanine crystals located on the surface of the rod led to high resistance of dosimeters to water and humidity. The 24 h soaking of irradiated dosimeters in liquid water-reduced EPR signals by 3-4% and by 2-3% for dosimeters with 27% and 9.8% of DL-α-alanine, respectively. Three month storage time of irradiated dosimeters in room conditions decreases EPR signal for ∼3%.

Gibberellin-regulated gene in the basal region of rice leaf sheath encodes basic helix-loop-helix transcription factor.

Science.gov (United States)

Komatsu, Setsuko; Takasaki, Hironori

2009-07-01

Genes regulated by gibberellin (GA) during leaf sheath elongation in rice seedlings were identified using the transcriptome approach. mRNA from the basal regions of leaf sheaths treated with GA3 was analyzed by high-coverage gene expression profiling. 33,004 peaks were detected, and 30 transcripts showed significant changes in the presence of GA3. Among these, basic helix-loop-helix transcription factor (AK073385) was significantly upregulated. Quantitative PCR analysis confirmed that expression of AK073385 was controlled by GA3 in a time- and dose-dependent manner. Basic helix-loop-helix transcription factor (AK073385) is therefore involved in the regulation of gene expression by GA3.

New optically active and thermally stable poly(amide-imide)s containing N,N'-(Bicyclo[2,2,2]oct-7-ene-2,3,5,6-tetracarboxylic)-bis-L-alanine and aromatic diamines: synthesis and characterization

Energy Technology Data Exchange (ETDEWEB)

Faghihi, Khalil; Absalar, Morteza; Hajibeygi, Mohsen [Arak University (Iran, Islamic Republic of). Faculty of Science. Organic Polymer Chemistry Research Lab.

2009-07-01

Five new optically active poly(amide-imide)s (PAIs) 6a-e were prepared by direct polycondensation reaction of the newly synthesized N,N'-(bicyclo[2,2,2]oct-7-ene-2,3,5,6-tetra carboxylic)-bis-L-alanine 4 with various aromatic diamines 5a-e using polar aprotic solvents such as N-methyl-2-pyrrolidone (NMP). In this technique triphenyl phosphite (TPP) and pyridine were used as condensing agents to form poly(amide-imide)s through the N-phosphonium salts of pyridine. All of the polymers were obtained in quantitative yields with inherent viscosities between 0.29-0.46 dL g{sup -1} and were highly soluble in polar aprotic solvents such as N,N-dimethyl acetamide (DMAc), N,N-dimethyl formamide (DMF), dimethyl sulfoxide (DMSO), N-methyl- 2-pyrrolidone (NMP) and solvents such as sulfuric acid. They were fully characterized by means of {sup 1}H NMR, FTIR spectroscopy, elemental analyses, inherent viscosity, solubility test, specific rotation and thermal properties of these polymers were investigated using thermogravimetric analysis techniques (TGA and DTG). (author)

Alanine-polymer dosemeter of ionizing radiation

International Nuclear Information System (INIS)

Tomasinski, Z.; Mirkowski, K.; Panta, P.; Stachowicz, W.

1994-01-01

The method of chemical preparation of alanine-copolymer of ethylene and vinyl acetate has been worked out. The material has been in a form of rods. The content of alanine has not exceeded 30%. The ESR signal of alanine radicals has been detected after exposition to ionizing radiation. The dose-response relationship has been presented

The vibrational spectrum of the hydrated alanine-leucine peptide in the amide region from IR experiments and first principles calculations

Science.gov (United States)

Hassan, Irtaza; Donati, Luca; Stensitzki, Till; Keller, Bettina G.; Heyne, Karsten; Imhof, Petra

2018-04-01

We have combined infrared (IR) experiments with molecular dynamics (MD) simulations in solution at finite temperature to analyse the vibrational signature of the small floppy peptide Alanine-Leucine. IR spectra computed from first-principles MD simulations exhibit no distinct differences between conformational clusters of α -helix or β -sheet-like folds with different orientations of the bulky leucine side chain. All computed spectra show two prominent bands, in good agreement with the experiment, that are assigned to the stretch vibrations of the carbonyl and carboxyl group, respectively. Variations in band widths and exact maxima are likely due to small fluctuations in the backbone torsion angles.

Alanine transaminase (ALT) blood test

Science.gov (United States)

... gov/ency/article/003473.htm Alanine transaminase (ALT) blood test To use the sharing features on this page, please enable JavaScript. The alanine transaminase (ALT) blood test measures the level of the enzyme ALT in ...

Structural studies of polypeptides: Mechanism of immunoglobin catalysis and helix propagation in hybrid sequence, disulfide containing peptides

Energy Technology Data Exchange (ETDEWEB)

Storrs, R.W.

1992-08-01

Catalytic immunoglobin fragments were studied Nuclear Magnetic Resonance spectroscopy to identify amino acid residues responsible for the catalytic activity. Small, hybrid sequence peptides were analyzed for helix propagation following covalent initiation and for activity related to the protein from which the helical sequence was derived. Hydrolysis of p-nitrophenyl carbonates and esters by specific immunoglobins is thought to involve charge complementarity. The pK of the transition state analog P-nitrophenyl phosphate bound to the immunoglobin fragment was determined by [sup 31]P-NMR to verify the juxtaposition of a positively charged amino acid to the binding/catalytic site. Optical studies of immunoglobin mediated photoreversal of cis, syn cyclobutane thymine dimers implicated tryptophan as the photosensitizing chromophore. Research shows the chemical environment of a single tryptophan residue is altered upon binding of the thymine dimer. This tryptophan residue was localized to within 20 [Angstrom] of the binding site through the use of a nitroxide paramagnetic species covalently attached to the thymine dimer. A hybrid sequence peptide was synthesized based on the bee venom peptide apamin in which the helical residues of apamin were replaced with those from the recognition helix of the bacteriophage 434 repressor protein. Oxidation of the disufide bonds occured uniformly in the proper 1-11, 3-15 orientation, stabilizing the 434 sequence in an [alpha]-helix. The glycine residue stopped helix propagation. Helix propagation in 2,2,2-trifluoroethanol mixtures was investigated in a second hybrid sequence peptide using the apamin-derived disulfide scaffold and the S-peptide sequence. The helix-stop signal previously observed was not observed in the NMR NOESY spectrum. Helical connectivities were seen throughout the S-peptide sequence. The apamin/S-peptide hybrid binded to the S-protein (residues 21-166 of ribonuclease A) and reconstituted enzymatic activity.

Double-helix stellarator

International Nuclear Information System (INIS)

Moroz, P.E.

1997-09-01

A new stellarator configuration, the Double-Helix Stellarator (DHS), is introduced. This novel configuration features a double-helix center post as the only helical element of the stellarator coil system. The DHS configuration has many unique characteristics. One of them is the extreme low plasma aspect ratio, A ∼ 1--1.2. Other advantages include a high enclosed volume, appreciable rotational transform, and a possibility of extreme-high-β MHD equilibria. Moreover, the DHS features improved transport characteristics caused by the absence of the magnetic field ripple on the outboard of the torus. Compactness, simplicity and modularity of the coil system add to the DHS advantages for fusion applications

Alanine metabolism in pyridoxine-depleted rat liver

International Nuclear Information System (INIS)

Okada, Mitsuko; Abe, Midori

1976-01-01

Alanine metabolism in normal and pyridoxine-deficient rats was studied in vivo and in vitro. Incorporation of 14 C-alanine into various liver components was determined and no difference was shown between normal and deficient animals in the incorporation into liver homogenates, lipid, protein and plasma glucose. Using the liver slice system, gluconeogenic activity from alanine or pyruvate was 40% lower in deficient rats compared with the activity of normal rats. However, inhibition was completely removed by the addition of 2-oxoglutarate to alanine. Penicillamine did not affect glucose formation from alanine in the liver slice. (auth.)

Effect of molecular weight on the quality of poly(alpha-methylstyrene mandrel

Directory of Open Access Journals (Sweden)

Xiuyun Shangguan

2017-07-01

Full Text Available Hollow poly(alpha-methylstyrene (PAMS shows application in inertial confinement fusion experiments as the degradable mandrels of glow plasma polymer shells. However, the molecular weight of PAMS has great influence on the quality of mandrels. In this work, this influence was systematically studied using several PAMS samples with different molecular weights. For PAMS shells with 900 μm inner diameter and different wall thickness, when the molecular weight of PAMS is in the range of 300–500 kg·mol−1, perfect sphericity and good wall thickness uniformity can be obtained. In contrast, when increasing molecular weight to 800 kg·mol−1, the sphericity and the wall thickness uniformity become worse. Moreover, compared with the wall uniformity, the sphericity of PAMS shells was much less sensitive to the molecular weight. The results also showed that the stability of W1/O compound droplets of PAMS shells were less affected by the molecular weight. It was revealed that the wall uniformity and the sphericity of the PAMS shells were associated with the diffusion rates of fluorobenzene (FB.

Protection against hyperthermic cell killing by alanine

International Nuclear Information System (INIS)

Cunningham, A.; Henle, K.J.; Moss, A.J.; Nagle, W.A.

1987-01-01

Compounds capable of protecting cells against hyperthermia may provide new insights into potential mechanisms of thermotolerance and cellular heat death. The authors characterized heat protection by alanine and related compounds as a function of concentration, temperature and preincubation time. Alanine was added either to complete medium or to HBSS before hyperthermia. Maximal heat protection required 3 hr, 37 0 ; longer preincubation intervals resulted in lower levels of protection. Addition of alanine to medium after hyperthermia had no protective effect. Protection was concentration dependent with a 20- or 200-fold increase in cell survival after 40 min, 45 0 C at 60 mM in medium or in HBSS, respectively. Higher alanine concentrations up to 120mM did not significantly increase heat protection. A 45 0 -heat survival curve showed that 100mM alanine increased the D/sub q/ by approx. 12 min with little change in the D/sub o/. Hyperthermia of 1 hr at temperatures between 42 0 and 45 0 indicated that 100mM alanine shifted the isotoxic temperature by 0.5 Celsius degrees. Polymers of either L or D,L alanine and related compounds, like pyruvate, also protected cells against heat killing. These results indicate that heat protection by alanine shows characteristics that are not shared by polyhydroxy compounds

Polymers with complexing properties. Simple poly(amino acids)

Science.gov (United States)

Roque, J. M.

1978-01-01

The free amino (0.3 equiv/residue) and carboxyl (0.5 equiv/residue) groups of thermal polylysine increased dramatically on treatment with distilled water. The total hydrolysis of such a polymer was abnormal in that only about 50% of the expected amino acids were recovered. Poly (lysine-co-alanine-co-glycine) under usual conditions hydrolyzed completely in 8 hours; whereas, when it was pretreated with diazomethane, a normal period of 24 hours was required to give (nearly) the same amounts of each free amino acid as compared with those obtained from the untreated polymer. The amino groups of the basic thermal poly(amino acids) were sterically hindered. The existence of nitrogen atoms linking two or three chains and reactive groups (anhydride, imine) were proposed.

Isotopic effects in mechanistic studies of biotransformations of fluorine derivatives of L-alanine catalysed by L-alanine dehydrogenase

International Nuclear Information System (INIS)

Szymańska-Majchrzak, Jolanta; Pałka, Katarzyna; Kańska, Marianna

2017-01-01

Synthesis of 3-fluoro-[2- 2 H]-L-alanine (3-F-[ 2 H]-L-Ala) in reductive amination of 3-fluoropyruvic acid catalysed by L-alanine dehydrogenase (AlaDH) was described. Fluorine derivative was used to study oxidative deamination catalysed by AlaDH applied kinetic (for 3-F-L-Ala in H 2 O - KIE’s on V max : 1.1; on V max /K M : 1.2; for 3-F-L-Ala in 2 H 2 O – on V max : 1.4; on V max /K M : 2.1) and solvent isotope effect methods (for 3-F-L-Ala - SIE’s on V max : 1.0; on V max /K M : 0.87; for 3-F-[2- 2 H]-L-Ala – on V max : 1.4; on V max /K M : 1.5). Studies explain some details of reaction mechanism. - Highlights: • Synthesis of 3-fluoro-[2- 2 H]-L-alanine was performed. • The reactions were catalysed using the enzyme L-alanine dehydrogenase. • Performed reactions involved fluorinated analogues of L-alanine. • Solvent isotope effects of deuterium were determined. • Kinetic isotope effects were determined for obtained 3-fluoro-L-alanine. • The mechanism of reaction catalysed by L-alanine dehydrogenase was proposed.

Crystallization and preliminary crystallographic analysis of d-alanine-d-alanine ligase from Streptococcus mutans

Energy Technology Data Exchange (ETDEWEB)

Lu, Yong-Zhi; Sheng, Yu [Institute for Nanobiomedical Technology and Membrane Biology, West China Hospital, Sichuan University, Chengdu 610065, Sichuan (China); Li, Lan-Fen [National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871 (China); Tang, De-Wei [Institute for Nanobiomedical Technology and Membrane Biology, West China Hospital, Sichuan University, Chengdu 610065, Sichuan (China); Liu, Xiang-Yu [National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871 (China); Zhao, Xiaojun, E-mail: zhaoxj@scu.edu.cn [Institute for Nanobiomedical Technology and Membrane Biology, West China Hospital, Sichuan University, Chengdu 610065, Sichuan (China); Liang, Yu-He, E-mail: zhaoxj@scu.edu.cn; Su, Xiao-Dong [National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871 (China); Institute for Nanobiomedical Technology and Membrane Biology, West China Hospital, Sichuan University, Chengdu 610065, Sichuan (China)

2007-09-01

A potential target for antibiotic drug design, d-alanine-d-alanine ligase from S. mutans, was expressed in E. coli, purified and crystallized. Diffraction data were collected to 2.4 Å resolution. d-Alanine-d-alanine ligase is encoded by the gene ddl (SMU-599) in Streptococcus mutans. This ligase plays a very important role in cell-wall biosynthesis and may be a potential target for drug design. To study the structure and function of this ligase, the gene ddl was amplified from S. mutans genomic DNA and cloned into the expression vector pET28a. The protein was expressed in soluble form in Escherichia coli strain BL21 (DE3). Homogeneous protein was obtained using a two-step procedure consisting of Ni{sup 2+}-chelating and size-exclusion chromatography. Purified protein was crystallized and the cube-shaped crystal diffracted to 2.4 Å. The crystal belongs to space group P3{sub 1}21 or P3{sub 2}21, with unit-cell parameters a = b = 79.50, c = 108.97 Å. There is one molecule per asymmetric unit.

Crystallization and preliminary crystallographic analysis of d-alanine-d-alanine ligase from Streptococcus mutans

International Nuclear Information System (INIS)

Lu, Yong-Zhi; Sheng, Yu; Li, Lan-Fen; Tang, De-Wei; Liu, Xiang-Yu; Zhao, Xiaojun; Liang, Yu-He; Su, Xiao-Dong

2007-01-01

A potential target for antibiotic drug design, d-alanine-d-alanine ligase from S. mutans, was expressed in E. coli, purified and crystallized. Diffraction data were collected to 2.4 Å resolution. d-Alanine-d-alanine ligase is encoded by the gene ddl (SMU-599) in Streptococcus mutans. This ligase plays a very important role in cell-wall biosynthesis and may be a potential target for drug design. To study the structure and function of this ligase, the gene ddl was amplified from S. mutans genomic DNA and cloned into the expression vector pET28a. The protein was expressed in soluble form in Escherichia coli strain BL21 (DE3). Homogeneous protein was obtained using a two-step procedure consisting of Ni 2+ -chelating and size-exclusion chromatography. Purified protein was crystallized and the cube-shaped crystal diffracted to 2.4 Å. The crystal belongs to space group P3 1 21 or P3 2 21, with unit-cell parameters a = b = 79.50, c = 108.97 Å. There is one molecule per asymmetric unit

The swimming of a perfect deforming helix

Science.gov (United States)

Koens, Lyndon; Zhang, Hang; Mourran, Ahmed; Lauga, Eric

2017-11-01

Many bacteria rotate helical flagellar filaments in order to swim. When at rest or rotated counter-clockwise these flagella are left handed helices but they undergo polymorphic transformations to right-handed helices when the motor is reversed. These helical deformations themselves can generate motion, with for example Rhodobacter sphaeroides using the polymorphic transformation of the flagellum to generate rotation, or Spiroplasma propagating a change of helix handedness across its body's length to generate forward motion. Recent experiments reported on an artificial helical microswimmer generating motion without a propagating change in handedness. Made of a temperature sensitive gel, these swimmers moved by changing the dimensions of the helix in a non-reciprocal way. Inspired by these results and helix's ubiquitous presence in the bacterial world, we investigate how a deforming helix moves within a viscous fluid. Maintaining a single handedness along its entire length, we discuss how a perfect deforming helix can create a non-reciprocal swimming stroke, identify its principle directions of motion, and calculate the swimming kinematics asymptotically.

Double-brush Langmuir-Blodgett monolayers of alpha-helical diblock copolypeptides

NARCIS (Netherlands)

Nguyen, Le-Thu T.; Vorenkamp, Eltjo J.; Daumont, Christophe J. M.; ten Brinke, Gerrit; Schouten, Arend J.; Vukovic, Ivana; Loos, Katja

2010-01-01

The synthesis of amphiphilic diblock copolypeptides consisting of poly(alpha-L-glutamic acid) (PLGA) and poly(gamma-methyl-L-glutamate-ran-gamma-stearyl-L-glutamate) with 30 mol % of stearyl substituents (PMLGSLG) and their monolayer behavior at the air-water interface have been studied.

Nanoparticles with entrapped {alpha}-tocopherol: synthesis, characterization, and controlled release

Energy Technology Data Exchange (ETDEWEB)

Zigoneanu, Imola Gabriela [101 E B Doran Building, BAE Department, Louisiana State University Agricultural Center, Baton Rouge, LA 70803 (United States); Astete, Carlos Ernesto [110 E B Doran Building, BAE Department, Louisiana State University Agricultural Center, Baton Rouge, LA 70803 (United States); Sabliov, Cristina Mirela [141 E B Doran Building, BAE Department, Louisiana State University Agricultural Center, Baton Rouge, LA 70803 (United States)], E-mail: csabliov@lsu.edu

2008-03-12

An emulsion evaporation method was used to synthesize spherical poly(DL-lactide-co-glycolide) (PLGA) nanoparticles with entrapped {alpha}-tocopherol. Two different surfactants were used: sodium dodecyl sulfate (SDS) and poly(vinyl alcohol) (PVA). For SDS nanoparticles, the size of the nanoparticles decreased significantly with the entrapment of {alpha}-tocopherol in the PLGA matrix, while the size of PVA nanoparticles remained unchanged. The polydispersity index after synthesis was under 0.100 for PVA nanoparticles and around 0.150 for SDS nanoparticles. The zeta potential was negative for all PVA nanoparticles. The entrapment efficiency of {alpha}-tocopherol in the polymeric matrix was approximately 89% and 95% for nanoparticles with 8% and 16% {alpha}-tocopherol theoretical loading, respectively. The residual PVA associated with the nanoparticles after purification was approximately 6% ( w/w relative to the nanoparticles). The release profile showed an initial burst followed by a slower release of the {alpha}-tocopherol entrapped inside the PLGA matrix. The release for nanoparticles with 8% {alpha}-tocopherol theoretical loading (86% released in the first hour) was faster than the release for the nanoparticles with 16% {alpha}-tocopherol theoretical loading (34% released in the first hour)

Alanine substitutions in the GXXXG motif alter C99 cleavage by γ-secretase but not its dimerization.

Science.gov (United States)

Higashide, Hidekazu; Ishihara, Seiko; Nobuhara, Mika; Ihara, Yasuo; Funamoto, Satoru

2017-03-01

The amyloid β (Aβ) protein is a major component of senile plaques, one of the neuropathological hallmarks of Alzheimer's disease. Amyloidogenic processing of amyloid precursor protein (APP) by β- and γ-secretases leads to production of Aβ. APP contains tandem triple repeats of the GXXXG motif in its extracellular juxtamembrane and transmembrane regions. It is reported that the GXXXG motif is related to protein-protein interactions, but it remains controversial whether the GXXXG motif in APP is involved in substrate dimerization and whether dimerization affects γ-secretase-dependent cleavage. Therefore, the relationship between the GXXXG motifs, substrate dimerization, and γ-secretase-dependent cleavage sites remains unclear. Here, we applied blue native poly acrylamide gel electrophoresis to examine the effect of alanine substitutions within the GXXXG motifs of APP carboxyl terminal fragment (C99) on its dimerization and Aβ production. Surprisingly, alanine substitutions in the motif failed to alter C99 dimerization in detergent soluble state. Cell-based and solubilized γ-secretase assays demonstrated that increasing alanine substitutions in the motif tended to decrease long Aβ species such as Aβ42 and Aβ43 and to increase in short Aβ species concomitantly. Our data suggest that the GXXXG motif is crucial for Aβ production, but not for C99 dimerization. © 2016 International Society for Neurochemistry.

Alanine increases blood pressure during hypotension

Science.gov (United States)

Conlay, L. A.; Maher, T. J.; Wurtman, R. J.

1990-01-01

The effect of L-alanine administration on blood pressure (BP) during haemorrhagic shock was investigated using anesthetized rats whose left carotid arteries were cannulated for BP measurement, blood removal, and drug administration. It was found that L-alanine, in doses of 10, 25, 50, 100, and 200 mg/kg, increased the systolic BP of hypotensive rats by 38 to 80 percent (while 100 mg/kg pyruvate increased BP by only 9.4 mmhg, not significantly different from saline). The results suggest that L-alanine might influence cardiovascular function.

Expression, crystallization and preliminary X-ray crystallographic analysis of Xoo0352, d-alanine-d-alanine ligase A, from Xanthomonas oryzae pv. oryzae

International Nuclear Information System (INIS)

Doan, Thanh Thi Ngoc; Kim, Jin-Kwang; Kim, Hyesoon; Ahn, Yeh-Jin; Kim, Jeong-Gu; Lee, Byoung-Moo; Kang, Lin-Woo

2008-01-01

Xoo0352, which encodes d-alanine-d-alanine ligase A (DdlA), from X. oryzae pv. oryzae was cloned, purified and crystallized. Preliminary X-ray crystallographic analysis of DdlA crystals was performed. Xanthomonas oryzae pv. oryzae (Xoo) causes bacterial blight (BB), which is one of the most devastating diseases of rice in most rice-growing countries. d-Alanine-d-alanine ligase A (DdlA), coded by the Xoo0352 gene, was expressed, purified and crystallized. DdlA is an enzyme that is involved in d-alanine metabolism and the biosynthesis of an essential bacterial peptidoglycan precursor, in which it catalyzes the formation of d-alanyl-d-alanine from two d-alanines, and is thus an attractive antibacterial drug target against Xoo. The DdlA crystals diffracted to 2.3 Å resolution and belonged to the primitive tetragonal space group P4 3 2 1 2, with unit-cell parameters a = b = 83.0, c = 97.6 Å. There is one molecule in the asymmetric unit, with a corresponding V M of 1.88 Å 3 Da −1 and a solvent content of 34.6%. The initial structure was determined by molecular replacement using d-alanine-d-alanine ligase from Staphylococcus aureus as a template model

Racemization of alanine by the alanine racemases from Salmonella typhimurium and Bacillus stearothermophilus: energetic reaction profiles

International Nuclear Information System (INIS)

Faraci, W.S.; Walsh, C.T.

1988-01-01

Alanine racemases are bacterial pyridoxal 5'-phosphate (PLP) dependent enzymes providing D-alanine as an essential building block for biosynthesis of the peptidoglycan layer of the cell wall. Two isozymic alanine racemases, encoded by the dadB gene and the alr gene, from the Gram-negative mesophilic Salmonella typhimurium and one from the Gram-positive thermophilic Bacillus stearothermophilus have been examined for the racemization mechanism. Substrate deuterium isotope effects and solvent deuterium isotope effects have been measured in both L → D and D→ L directions for all three enzymes to assess the degree to which abstraction of the α-proton or protonation of substrate PLP carbanion is limiting in catalysis. Additionally, experiments measuring internal return of α- 3 H from substrate to product and solvent exchange/substrate conversion experiments in 3 H 2 O have been used with each enzyme to examine the partitioning of substrate PLP carbanion intermediates and to obtain the relative heights of kinetically significant energy barriers in alanine racemase catalysis

New insight into the solution structures of wheat gluten proteins from Raman optical activity

DEFF Research Database (Denmark)

Blanch, E.W.; Kasarda, D.D.; Hecht, L.

2003-01-01

Vibrational Raman optical activity (ROA) spectra of the wheat proteins a-gliadin (A-gliadin), omega-liadin, and a 30 kDa peptide called T-A-1 from the high molecular weight glutenin subunit (HMW-GS) Dx5 were measured to obtain new information about their solution structures. The spectral data show...... that, under the conditions investigated, A-gliadin contains a considerable amount of hydrated alpha-helix, most of which probably lies within a relatively structured C-terminal domain. Smaller quantities of beta-structure and poly(L-proline) II (PPII) helix were also identified. Addition of methanol...

Imaging analysis of direct alanine uptake by rice seedlings

International Nuclear Information System (INIS)

Nihei, Naoto; Masuda, Sayaka; Rai, Hiroki; Nakanishi, Tomoko M.

2008-01-01

We presented alanine, a kind of amino acids, uptake by a rice seedling to study the basic mechanism of the organic fertilizer effectiveness in organic farming. The rice grown in the culture solution containing alanine as a nitrogen source absorbed alanine approximately two times faster than that grown with NH 4 + from analysis of 14 C-alanine images by Imaging Plate method. It was suggested that the active transport ability of the rice seeding was induced in roots by existence of alanine in the rhizosphere. The alanine uptake images of the rice roots were acquired every 5 minutes successively by the real-time autoradiography system we developed. The analysis of the successive images showed that alanine uptake was not uniform throughout the root but especially active at the root tip. (author)

Designing cooperatively folded abiotic uni- and multimolecular helix bundles

Science.gov (United States)

de, Soumen; Chi, Bo; Granier, Thierry; Qi, Ting; Maurizot, Victor; Huc, Ivan

2018-01-01

Abiotic foldamers, that is foldamers that have backbones chemically remote from peptidic and nucleotidic skeletons, may give access to shapes and functions different to those of peptides and nucleotides. However, design methodologies towards abiotic tertiary and quaternary structures are yet to be developed. Here we report rationally designed interactional patterns to guide the folding and assembly of abiotic helix bundles. Computational design facilitated the introduction of hydrogen-bonding functionalities at defined locations on the aromatic amide backbones that promote cooperative folding into helix-turn-helix motifs in organic solvents. The hydrogen-bond-directed aggregation of helices not linked by a turn unit produced several thermodynamically and kinetically stable homochiral dimeric and trimeric bundles with structures that are distinct from the designed helix-turn-helix. Relative helix orientation within the bundles may be changed from parallel to tilted on subtle solvent variations. Altogether, these results prefigure the richness and uniqueness of abiotic tertiary structure behaviour.

Genome-wide identification of estrogen receptor alpha-binding sites in mouse liver

DEFF Research Database (Denmark)

Gao, Hui; Fält, Susann; Sandelin, Albin

2007-01-01

We report the genome-wide identification of estrogen receptor alpha (ERalpha)-binding regions in mouse liver using a combination of chromatin immunoprecipitation and tiled microarrays that cover all nonrepetitive sequences in the mouse genome. This analysis identified 5568 ERalpha-binding regions...... genes. The majority of ERalpha-binding regions lie in regions that are evolutionarily conserved between human and mouse. Motif-finding algorithms identified the estrogen response element, and variants thereof, together with binding sites for activator protein 1, basic-helix-loop-helix proteins, ETS...... signaling in mouse liver, by characterizing the first step in this signaling cascade, the binding of ERalpha to DNA in intact chromatin....

Mirrors in the PDB: left-handed alpha-turns guide design with D-amino acids.

Science.gov (United States)

Annavarapu, Srinivas; Nanda, Vikas

2009-09-22

Incorporating variable amino acid stereochemistry in molecular design has the potential to improve existing protein stability and create new topologies inaccessible to homochiral molecules. The Protein Data Bank has been a reliable, rich source of information on molecular interactions and their role in protein stability and structure. D-amino acids rarely occur naturally, making it difficult to infer general rules for how they would be tolerated in proteins through an analysis of existing protein structures. However, protein elements containing short left-handed turns and helices turn out to contain useful information. Molecular mechanisms used in proteins to stabilize left-handed elements by L-amino acids are structurally enantiomeric to potential synthetic strategies for stabilizing right-handed elements with D-amino acids. Propensities for amino acids to occur in contiguous alpha(L) helices correlate with published thermodynamic scales for incorporation of D-amino acids into alpha(R) helices. Two backbone rules for terminating a left-handed helix are found: an alpha(R) conformation is disfavored at the amino terminus, and a beta(R) conformation is disfavored at the carboxy terminus. Helix capping sidechain-backbone interactions are found which are unique to alpha(L) helices including an elevated propensity for L-Asn, and L-Thr at the amino terminus and L-Gln, L-Thr and L-Ser at the carboxy terminus. By examining left-handed alpha-turns containing L-amino acids, new interaction motifs for incorporating D-amino acids into right-handed alpha-helices are identified. These will provide a basis for de novo design of novel heterochiral protein folds.

Spatio-temporal profiling and degradation of alpha-amylase isozymes during barley seed germination

DEFF Research Database (Denmark)

Bak-Jensen, K.S.; Laugesen, Sabrina; Østergaard, Ole

2007-01-01

Ten genes from two multigene families encode barley alpha-amylases. To gain insight into the occurrence and fate of individual isoforms during seed germination, the alpha-amylase repertoire was mapped by using a proteomics approach consisting of 2D gel electrophoresis, western blotting, and mass...... increased during germination. Assessing the fragment minimum chain length by peptide mass fingerprinting suggested that alpha-amylase 2 ( gi vertical bar 4699831) initially was cleaved just prior to domain B that protrudes from the (beta alpha)(8)-barrel between beta-strand 3 and alpha-helix 3, followed...... essentially only full-length alpha-amylase forms. While only products of the above three genes appeared by germination also of 15 other barley cultivars, the cultivars had distinct repertoires of charge and molecular mass variant forms. These patterns appeared not to be correlated with malt quality....

Alanine metabolism in acute falciparum malaria

NARCIS (Netherlands)

Pukrittayakamee, S.; Krishna, S.; ter Kuile, F.; Wilaiwan, O.; Williamson, D. H.; White, N. J.

2002-01-01

We investigated the integrity of the gluconeogenic pathway in severe malaria using alanine metabolism as a measure. Alanine disposition and liver blood flow, assessed by indocyanine green (ICG) clearance, were measured simultaneously in 10 patients with falciparum malaria (six severe and four

Alanine EPR dosimetry of therapeutic irradiators

International Nuclear Information System (INIS)

Bugay, O.; Bartchuk, V.; Kolesnik, S.; Mazin, M.; Gaponenko, H.

1999-01-01

The high-dose alanine EPR dosimetry is a very precise method in the dose range 1-100 kGy. The system is used generally as the standard high-dose transfer dosimetry in many laboratories. This is comparatively expensive technique so it is important to use it as a more universal dosimetry system also in the middle and low dose ranges. The problems of the middle-dose alanine dosimetry are discussed and the solution of several problems is proposed. The alanine EPR dosimetry has been applied to the dose measurements of medical irradiators in the Kiev City Oncology Center. (author)

Study on the EPR/dosimetric properties of some substituted alanines

International Nuclear Information System (INIS)

Gancheva, Veselka; Sagstuen, Einar; Yordanov, Nicola D.

2006-01-01

Polycrystalline phenyl-alanine and perdeuterated l-α-alanine (l-α-alanine-d 4 ) were studied as potential high-energy radiation-sensitive materials (RSM) for solid state/EPR dosimetry. It was found that phenyl-alanine exhibits a linear dose response in the dose region 0.1-17kGy. However, phenyl-alanine is about 10 times less sensitive to γ-irradiation than standard l-α-alanine irradiated at the same doses. Moreover, the EPR response from phenyl-alanine is unstable and, independent of the absorbed dose, decreases by about 50% within 20 days after irradiation upon storage at room temperature. γ-irradiated polycrystalline perdeuterated l-α-alanine (CD 3 CD(NH 2 )COOH) has not previously been studied at room temperature by EPR spectroscopy. The first part of the present analysis was with respect to the structure of the EPR spectrum. By spectrum simulations, the presence of at least two radiation induced free radicals, R 1 =CH 3 C*(H)COOH and R 2 =H 3 N + -C*(CH 3 )COO - , was confirmed very clearly. Both these radicals were suggested previously from EPR and ENDOR studies of standard alanine crystals. The further investigations into the potential use of alanine-d 4 as RSM, after choosing optimal EPR spectrometer settings parameters for this purpose, show that it is ca. two times more sensitive than standard l-α-alanine

Medical reference dosimetry using EPR measurements of alanine

DEFF Research Database (Denmark)

Helt-Hansen, Jakob; Rosendal, F.; Kofoed, I.M.

2009-01-01

Background. Electron spin resonance (EPR) is used to determine the absorbed dose of alanine dosimeters exposed to clinical photon beams in a solid-water phantom. Alanine is potentially suitable for medical reference dosimetry, because of its near water equivalence over a wide energy spectrum, low...... methods the proposed algorithm can be applied without normalisation of phase shifts caused by changes in the g-value of the cavity. The study shows that alanine dosimetry is a suitable candidate for medical reference dosimetry especially for quality control applications.......Background. Electron spin resonance (EPR) is used to determine the absorbed dose of alanine dosimeters exposed to clinical photon beams in a solid-water phantom. Alanine is potentially suitable for medical reference dosimetry, because of its near water equivalence over a wide energy spectrum, low...

Helix probe areas for the utilization of geothermal power. A practical example; Helix-Sondenfelder zur Nutzung von Erdwaerme. Ein Praxisbeispiel

Energy Technology Data Exchange (ETDEWEB)

Kuebert, Markus; Walker-Hertkorn, Simone [tewag Technologie - Erdwaermeanlagen - Umweltschutz GmbH, Starzach (Germany); Tietz, Jan [REHAU AG und Co., Erlangen-Eltersdorf (Germany); Riepold, Markus; Gloeckl, Andreas [MR Tiefbau GmbH, Brunnen (Germany)

2013-02-01

Thanks to their spiral shape so-called helix probes with a tube length of 40 meter have a height of only three meter: A lot of heat exchange area in a small space. Thus, helix probes are an ideal solution for the utilization of geothermal energy at places at which one cannot drill deeply due to geothermal reasons. Under this aspect, the contribution under consideration reports on the planning of a helix probe area being sustainably adapted to the user requirements for the new construction of a production facility.

Alanine racemase mutants of Burkholderia pseudomallei and Burkholderia mallei and use of alanine racemase as a non-antibiotic-based selectable marker.

Directory of Open Access Journals (Sweden)

Sheryl L W Zajdowicz

Full Text Available Burkholderia pseudomallei and Burkholderia mallei are category B select agents and must be studied under BSL3 containment in the United States. They are typically resistant to multiple antibiotics, and the antibiotics used to treat B. pseudomallei or B. mallei infections may not be used as selective agents with the corresponding Burkholderia species. Here, we investigated alanine racemase deficient mutants of B. pseudomallei and B. mallei for development of non-antibiotic-based genetic selection methods and for attenuation of virulence. The genome of B. pseudomallei K96243 has two annotated alanine racemase genes (bpsl2179 and bpss0711, and B. mallei ATCC 23344 has one (bma1575. Each of these genes encodes a functional enzyme that can complement the alanine racemase deficiency of Escherichia coli strain ALA1. Herein, we show that B. pseudomallei with in-frame deletions in both bpsl2179 and bpss0711, or B. mallei with an in-frame deletion in bma1575, requires exogenous D-alanine for growth. Introduction of bpsl2179 on a multicopy plasmid into alanine racemase deficient variants of either Burkholderia species eliminated the requirement for D-alanine. During log phase growth without D-alanine, the viable counts of alanine racemase deficient mutants of B. pseudomallei and B. mallei decreased within 2 hours by about 1000-fold and 10-fold, respectively, and no viable bacteria were present at 24 hours. We constructed several genetic tools with bpsl2179 as a selectable genetic marker, and we used them without any antibiotic selection to construct an in-frame ΔflgK mutant in the alanine racemase deficient variant of B. pseudomallei K96243. In murine peritoneal macrophages, wild type B. mallei ATCC 23344 was killed much more rapidly than wild type B. pseudomallei K96243. In addition, the alanine racemase deficient mutant of B. pseudomallei K96243 exhibited attenuation versus its isogenic parental strain with respect to growth and survival in murine

Alanine Racemase Mutants of Burkholderia pseudomallei and Burkholderia mallei and Use of Alanine Racemase as a Non-Antibiotic-Based Selectable Marker

Science.gov (United States)

Zajdowicz, Sheryl L. W.; Jones-Carson, Jessica; Vazquez-Torres, Andres; Jobling, Michael G.; Gill, Ronald E.; Holmes, Randall K.

2011-01-01

Burkholderia pseudomallei and Burkholderia mallei are category B select agents and must be studied under BSL3 containment in the United States. They are typically resistant to multiple antibiotics, and the antibiotics used to treat B. pseudomallei or B. mallei infections may not be used as selective agents with the corresponding Burkholderia species. Here, we investigated alanine racemase deficient mutants of B. pseudomallei and B. mallei for development of non-antibiotic-based genetic selection methods and for attenuation of virulence. The genome of B. pseudomallei K96243 has two annotated alanine racemase genes (bpsl2179 and bpss0711), and B. mallei ATCC 23344 has one (bma1575). Each of these genes encodes a functional enzyme that can complement the alanine racemase deficiency of Escherichia coli strain ALA1. Herein, we show that B. pseudomallei with in-frame deletions in both bpsl2179 and bpss0711, or B. mallei with an in-frame deletion in bma1575, requires exogenous d-alanine for growth. Introduction of bpsl2179 on a multicopy plasmid into alanine racemase deficient variants of either Burkholderia species eliminated the requirement for d-alanine. During log phase growth without d-alanine, the viable counts of alanine racemase deficient mutants of B. pseudomallei and B. mallei decreased within 2 hours by about 1000-fold and 10-fold, respectively, and no viable bacteria were present at 24 hours. We constructed several genetic tools with bpsl2179 as a selectable genetic marker, and we used them without any antibiotic selection to construct an in-frame ΔflgK mutant in the alanine racemase deficient variant of B. pseudomallei K96243. In murine peritoneal macrophages, wild type B. mallei ATCC 23344 was killed much more rapidly than wild type B. pseudomallei K96243. In addition, the alanine racemase deficient mutant of B. pseudomallei K96243 exhibited attenuation versus its isogenic parental strain with respect to growth and survival in murine peritoneal macrophages

A conserved degron containing an amphipathic helix regulates the cholesterol-mediated turnover of human squalene monooxygenase, a rate-limiting enzyme in cholesterol synthesis.

Science.gov (United States)

Chua, Ngee Kiat; Howe, Vicky; Jatana, Nidhi; Thukral, Lipi; Brown, Andrew J

2017-12-08

Cholesterol biosynthesis in the endoplasmic reticulum (ER) is tightly controlled by multiple mechanisms to regulate cellular cholesterol levels. Squalene monooxygenase (SM) is the second rate-limiting enzyme in cholesterol biosynthesis and is regulated both transcriptionally and post-translationally. SM undergoes cholesterol-dependent proteasomal degradation when cholesterol is in excess. The first 100 amino acids of SM (designated SM N100) are necessary for this degradative process and represent the shortest cholesterol-regulated degron identified to date. However, the fundamental intrinsic characteristics of this degron remain unknown. In this study, we performed a series of deletions, point mutations, and domain swaps to identify a 12-residue region (residues Gln-62-Leu-73), required for SM cholesterol-mediated turnover. Molecular dynamics and circular dichroism revealed an amphipathic helix within this 12-residue region. Moreover, 70% of the variation in cholesterol regulation was dependent on the hydrophobicity of this region. Of note, the earliest known Doa10 yeast degron, Deg1, also contains an amphipathic helix and exhibits 42% amino acid similarity with SM N100. Mutating SM residues Phe-35/Ser-37/Leu-65/Ile-69 into alanine, based on the key residues in Deg1, blunted SM cholesterol-mediated turnover. Taken together, our results support a model whereby the amphipathic helix in SM N100 attaches reversibly to the ER membrane depending on cholesterol levels; with excess, the helix is ejected and unravels, exposing a hydrophobic patch, which then serves as a degradation signal. Our findings shed new light on the regulation of a key cholesterol synthesis enzyme, highlighting the conservation of critical degron features from yeast to humans. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

On the existence of 'L-alanine cadmium bromide'.

Science.gov (United States)

Srinivasan, Bikshandarkoil R

2013-12-01

It is argued that the recently reported nonlinear optical crystal L-alanine cadmium bromide, grown by slow solvent evaporation method at room temperature [P. Ilayabarathi, J. Chandrasekaran, Spectrochim. Acta 96A (2012) 684-689] is the well-known L-alanine crystal. The isolation of L-alanine crystal is explained due to fractional crystallization. Copyright © 2013 Elsevier B.V. All rights reserved.

Solved? The reductive radiation chemistry of alanine.

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Pauwels, Ewald; De Cooman, Hendrik; Waroquier, Michel; Hole, Eli O; Sagstuen, Einar

2014-02-14

The structural changes throughout the entire reductive radiation-induced pathway of l-α-alanine are solved on an atomistic level with the aid of periodic DFT and nudged elastic band (NEB) simulations. This yields unprecedented information on the conformational changes taking place, including the protonation state of the carboxyl group in the "unstable" and "stable" alanine radicals and the internal transformation converting these two radical variants at temperatures above 220 K. The structures of all stable radicals were verified by calculating EPR properties and comparing those with experimental data. The variation of the energy throughout the full radiochemical process provides crucial insight into the reason why these structural changes and rearrangements occur. Starting from electron capture, the excess electron quickly localizes on the carbon of a carboxyl group, which pyramidalizes and receives a proton from the amino group of a neighboring alanine molecule, forming a first stable radical species (up to 150 K). In the temperature interval 150-220 K, this radical deaminates and deprotonates at the carboxyl group, the detached amino group undergoes inversion and its methyl group sustains an internal rotation. This yields the so-called "unstable alanine radical". Above 220 K, triggered by the attachment of an additional proton on the detached amino group, the radical then undergoes an internal rotation in the reverse direction, giving rise to the "stable alanine radical", which is the final stage in the reductive radiation-induced decay of alanine.

Interactions between the mixotrophic dinoflagellate Takayama helix and common heterotrophic protists.

Science.gov (United States)

Ok, Jin Hee; Jeong, Hae Jin; Lim, An Suk; Lee, Kyung Ha

2017-09-01

The phototrophic dinoflagellate Takayama helix that is known to be harmful to abalone larvae has recently been revealed to be mixotrophic. Although mixotrophy elevates the growth rate of T. helix by 79%-185%, its absolute growth rate is still as low as 0.3d -1 . Thus, if the mortality rate of T. helix due to predation is high, this dinoflagellate may not easily prevail. To investigate potential effective protistan grazers on T. helix, feeding by diverse heterotrophic dinoflagellates such as engulfment-feeding Oxyrrhis marina, Gyrodinium dominans, Gyrodinium moestrupii, Polykrikos kofoidii, and Noctiluca scintillans, peduncle-feeding Aduncodinium glandula, Gyrodiniellum shiwhaense, Luciella masanensis, and Pfiesteria piscicida, pallium-feeding Oblea rotunda and Protoperidinium pellucidum, and the naked ciliates Pelagostrobilidium sp. (ca. 40μm in cell length) and Strombidinopsis sp. (ca. 150μm in cell length) on T. helix was explored. Among the tested heterotrophic protists, O. marina, G. dominans, G. moestrupii, A. glandula, L. masanensis, P. kofoidii, P. piscicida, and Strombidinopsis sp. were able to feed on T. helix. The growth rates of all these predators except Strombidinopsis sp. with T. helix prey were lower than those without the prey. The growth rate of Strombidinopsis sp. on T. helix was almost zero although the growth rate of Strombidinopsis sp. with T. helix prey was higher than those without the prey. Moreover, T. helix fed on O. marina and P. pellucidum and lysed the cells of P. kofoidii and G. shiwhaense. With increasing the concentrations of T. helix, the growth rates of O. marina and P. kofoidii decreased, but those of G. dominans and L. masanensis largely did not change. Therefore, reciprocal predation, lysis, no feeding, and the low ingestion rates of the common protists preying on T. helix may result in a low mortality rate due to predation, thereby compensating for this species' low growth rate. Copyright © 2017 Elsevier B.V. All rights

Comparative Study on the Adaptation and Growth Dynamics of the Helix pomatia and Helix aspersa Muller Terrestrial Snails Under Different Feeding Regimes

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Adrian Toader-Williams

2010-05-01

Full Text Available We used Helix pomatia and Helix aspersa species and measure their growth as the snails were approaching the hibernation season. Helix pomatia 2yo shown a decrease in weight while being raised in enclosed parcels of 4sqm the younger Helix pomatia 1yo as well as Helix aspersa Muller demonstrated the ability to adapt relatively fast to the same conditions. We established 5 experimental lots in a Helix pomatia farm, GPS coordinates N46.606040 E23.599950. Control lot contained Taraxacum officinales, Sonchus oleraceus, Equisetum arvense and Atriplex hortensis, wild flora found within the farm. The other lots contained the same plants as the control lot plus different combinations of imported plants from other areals. The H. pomatia 2yo weight decreased in the control lot by a mean of -3.86% while H. aspersa 1yo marked an increase of +16.89% in the same lot during the same period. The lot containing lupinus polyphyllus delivered snails with weight gain of +24.66% for H. pomatia 2yo and an increase of only +1.98% for H. aspersa 1yo. As a contrast, H. pomatia 2yo gained only +7.72% while H. aspersa 1yo gained +28.89%, in the lot containing Lavanda officinalis, Foeniculum vulgare and Hyssopus officinalis among the other plants.

Government and Governance of Regional Triple Helix Interactions

Science.gov (United States)

Danson, Mike; Todeva, Emanuela

2016-01-01

This conceptual paper contributes to the discussion of the role of regional government and regional Triple Helix constellations driving economic development and growth within regional boundaries. The impact of regionalism and subsidiarity on regional Triple Helix constellations, and the questions of governmentality, governance and institutional…

A basic helix-loop-helix transcription factor, PhFBH4, regulates flower senescence by modulating ethylene biosynthesis pathway in petunia

Science.gov (United States)

The basic helix-loop-helix (bHLH) transcription factors (TFs) play important roles in regulating multiple biological processes in plants. However, there are few reports about the function of bHLHs in flower senescence. In this study, a bHLH TF, PhFBH4, was found to be dramatically upregulated during...

Mechanical properties and osteocompatibility of novel biodegradable alanine based polyphosphazenes: Side group effects.

Science.gov (United States)

Sethuraman, Swaminathan; Nair, Lakshmi S; El-Amin, Saadiq; Nguyen, My-Tien; Singh, Anurima; Krogman, Nick; Greish, Yaser E; Allcock, Harry R; Brown, Paul W; Laurencin, Cato T

2010-06-01

The versatility of polymers for tissue regeneration lies in the feasibility to modulate the physical and biological properties by varying the side groups grafted to the polymers. Biodegradable polyphosphazenes are high-molecular-weight polymers with alternating nitrogen and phosphorus atoms in the backbone. This study is the first of its kind to systematically investigate the effect of side group structure on the compressive strength of novel biodegradable polyphosphazene based polymers as potential materials for tissue regeneration. The alanine polyphosphazene based polymers, poly(bis(ethyl alanato) phosphazene) (PNEA), poly((50% ethyl alanato) (50% methyl phenoxy) phosphazene) (PNEA(50)mPh(50)), poly((50% ethyl alanato) (50% phenyl phenoxy) phosphazene) (PNEA(50)PhPh(50)) were investigated to demonstrate their mechanical properties and osteocompatibility. Results of mechanical testing studies demonstrated that the nature and the ratio of the pendent groups attached to the polymer backbone play a significant role in determining the mechanical properties of the resulting polymer. The compressive strength of PNEA(50)PhPh(50) was significantly higher than poly(lactide-co-glycolide) (85:15 PLAGA) (p<0.05). Additional studies evaluated the cellular response and gene expression of primary rat osteoblast cells on PNEA, PNEA(50)mPh(50) and PNEA(50)PhPh(50) films as candidates for bone tissue engineering applications. Results of the in vitro osteocompatibility evaluation demonstrated that cells adhere, proliferate, and maintain their phenotype when seeded directly on the surface of PNEA, PNEA(50)mPh(50), and PNEA(50)PhPh(50). Moreover, cells on the surface of the polymers expressed type I collagen, alkaline phosphatase, osteocalcin, osteopontin, and bone sialoprotein, which are characteristic genes for osteoblast maturation, differentiation, and mineralization. Copyright 2009 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Double helix vortex breakdown in a turbulent swirling annular jet flow

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Vanierschot, M.; Percin, M.; van Oudheusden, B. W.

2018-03-01

In this paper, we report on the structure and dynamics of double helix vortex breakdown in a turbulent annular swirling jet. Double helix breakdown has been reported previously for the laminar flow regime, but this structure has rarely been observed in turbulent flow. The flow field is investigated experimentally by means of time-resolved tomographic particle image velocimetry. Notwithstanding the axisymmetric nature of the time-averaged flow, analysis of the instantaneous three-dimensional (3D) vortical structures shows the existence of a vortex core along the central axis which breaks up into a double helix downstream. The winding sense of this double helix is opposite to the swirl direction (m =-2 ) and it is wrapped around a central vortex breakdown bubble. This structure is quite different from double helix breakdown found in laminar flows where the helix is formed in the wake of the bubble and not upstream. The double helix precesses around the central axis of the jet with a precessing frequency corresponding to a Strouhal number of 0.27.

Alanine-ESR dosimetry for radiotherapy IAEA experience

International Nuclear Information System (INIS)

Mehta, K.; Girzikowsky, R.; )

1997-01-01

At present, the most commonly used transfer dosimeters for radiotherapy applications are TL dosemeters. They are being used for intercomparison between SSDLs (about 70) and the IAEA dosimetry laboratory. However, there are some undesirable characteristics of this dosimetry system. We have a study in progress at the IAEA to evaluate the alanine-ESR systems as an alternative to TLDs. There are several desirable qualities which make alanine an attractive dosemeter. Preliminary data suggest that the alanine-ESR dosimetry system has the potential to replace TLDs for intercomparison amongst SSDLs in the therapy-level dose regions. (Author)

An α-Helix-Mimicking 12,13-Helix: Designed α/β/γ-Foldamers as Selective Inhibitors of Protein-Protein Interactions.

Science.gov (United States)

Grison, Claire M; Miles, Jennifer A; Robin, Sylvie; Wilson, Andrew J; Aitken, David J

2016-09-05

A major current challenge in bioorganic chemistry is the identification of effective mimics of protein secondary structures that act as inhibitors of protein-protein interactions (PPIs). In this work, trans-2-aminocyclobutanecarboxylic acid (tACBC) was used as the key β-amino acid component in the design of α/β/γ-peptides to structurally mimic a native α-helix. Suitably functionalized α/β/γ-peptides assume an α-helix-mimicking 12,13-helix conformation in solution, exhibit enhanced proteolytic stability in comparison to the wild-type α-peptide parent sequence from which they are derived, and act as selective inhibitors of the p53/hDM2 interaction. © 2016 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.

Uptake of neutral alpha- and beta-amino acids by human proximal tubular cells

DEFF Research Database (Denmark)

Jessen, H; Røigaard, H; Jacobsen, Christian

1996-01-01

experiments revealed that all the neutral amino acids tested reduced the uptake of AIB, whereas there was no effect of taurine, L-aspartic acid, and L-arginine. By contrast, the influx of beta-alanine was only drastically reduced by beta-amino acids, whereas the inhibition by neutral alpha-amino acids...

Applying Unique Molecular Identifiers in Next Generation Sequencing Reveals a Constrained Viral Quasispecies Evolution under Cross-Reactive Antibody Pressure Targeting Long Alpha Helix of Hemagglutinin

Science.gov (United States)

Hauck, Nastasja C.; Kirpach, Josiane; Kiefer, Christina; Farinelle, Sophie; Morris, Stephen A.; Muller, Claude P.; Lu, I-Na

2018-01-01

To overcome yearly efforts and costs for the production of seasonal influenza vaccines, new approaches for the induction of broadly protective and long-lasting immune responses have been developed in the past decade. To warrant safety and efficacy of the emerging crossreactive vaccine candidates, it is critical to understand the evolution of influenza viruses in response to these new immune pressures. Here we applied unique molecular identifiers in next generation sequencing to analyze the evolution of influenza quasispecies under in vivo antibody pressure targeting the hemagglutinin (HA) long alpha helix (LAH). Our vaccine targeting LAH of hemagglutinin elicited significant seroconversion and protection against homologous and heterologous influenza virus strains in mice. The vaccine not only significantly reduced lung viral titers, but also induced a well-known bottleneck effect by decreasing virus diversity. In contrast to the classical bottleneck effect, here we showed a significant increase in the frequency of viruses with amino acid sequences identical to that of vaccine targeting LAH domain. No escape mutant emerged after vaccination. These results not only support the potential of a universal influenza vaccine targeting the conserved LAH domains, but also clearly demonstrate that the well-established bottleneck effect on viral quasispecies evolution does not necessarily generate escape mutants. PMID:29587397

Applying Unique Molecular Identifiers in Next Generation Sequencing Reveals a Constrained Viral Quasispecies Evolution under Cross-Reactive Antibody Pressure Targeting Long Alpha Helix of Hemagglutinin

Directory of Open Access Journals (Sweden)

Nastasja C. Hauck

2018-03-01

Full Text Available To overcome yearly efforts and costs for the production of seasonal influenza vaccines, new approaches for the induction of broadly protective and long-lasting immune responses have been developed in the past decade. To warrant safety and efficacy of the emerging crossreactive vaccine candidates, it is critical to understand the evolution of influenza viruses in response to these new immune pressures. Here we applied unique molecular identifiers in next generation sequencing to analyze the evolution of influenza quasispecies under in vivo antibody pressure targeting the hemagglutinin (HA long alpha helix (LAH. Our vaccine targeting LAH of hemagglutinin elicited significant seroconversion and protection against homologous and heterologous influenza virus strains in mice. The vaccine not only significantly reduced lung viral titers, but also induced a well-known bottleneck effect by decreasing virus diversity. In contrast to the classical bottleneck effect, here we showed a significant increase in the frequency of viruses with amino acid sequences identical to that of vaccine targeting LAH domain. No escape mutant emerged after vaccination. These results not only support the potential of a universal influenza vaccine targeting the conserved LAH domains, but also clearly demonstrate that the well-established bottleneck effect on viral quasispecies evolution does not necessarily generate escape mutants.

IFITM3 requires an amphipathic helix for antiviral activity.

Science.gov (United States)

Chesarino, Nicholas M; Compton, Alex A; McMichael, Temet M; Kenney, Adam D; Zhang, Lizhi; Soewarna, Victoria; Davis, Matthew; Schwartz, Olivier; Yount, Jacob S

2017-10-01

Interferon-induced transmembrane protein 3 (IFITM3) is a cellular factor that blocks virus fusion with cell membranes. IFITM3 has been suggested to alter membrane curvature and fluidity, though its exact mechanism of action is unclear. Using a bioinformatic approach, we predict IFITM3 secondary structures and identify a highly conserved, short amphipathic helix within a hydrophobic region of IFITM3 previously thought to be a transmembrane domain. Consistent with the known ability of amphipathic helices to alter membrane properties, we show that this helix and its amphipathicity are required for the IFITM3-dependent inhibition of influenza virus, Zika virus, vesicular stomatitis virus, Ebola virus, and human immunodeficiency virus infections. The homologous amphipathic helix within IFITM1 is also required for the inhibition of infection, indicating that IFITM proteins possess a conserved mechanism of antiviral action. We further demonstrate that the amphipathic helix of IFITM3 is required to block influenza virus hemagglutinin-mediated membrane fusion. Overall, our results provide evidence that IFITM proteins utilize an amphipathic helix for inhibiting virus fusion. © 2017 The Authors.

Effect of Hedera helix on lung histopathology in chronic asthma.

Science.gov (United States)

Hocaoglu, Arzu Babayigit; Karaman, Ozkan; Erge, Duygu Olmez; Erbil, Guven; Yilmaz, Osman; Kivcak, Bijen; Bagriyanik, H Alper; Uzuner, Nevin

2012-12-01

Hedera helix is widely used to treat bronchial asthma for many years. However, effects of this herb on lung histopathology is still far from clear. We aimed to determine the effect of oral administration of Hedera helix on lung histopathology in a murine model of chronic asthma.BALB/c mice were divided into four groups; I (Placebo), II (Hedera helix), III (Dexamethasone) and IV (Control). All mice except controls were sensitized and challenged with ovalbumin. Then, mice in group I received saline, group II 100 mg/kg Hedera helix and group III 1 mg/kg dexamethasone via orogastic gavage once daily for one week. Airway histopathology was evaluated by using light and electron microscopy in all groups.Goblet cell numbers and thicknesses of basement membrane were found significantly lower in group II, but there was no statistically significant difference in terms of number of mast cells, thicknesses of epithelium and subepithelial smooth muscle layers between group I and II. When Hedera helix and dexamethasone groups were compared with each other, thickness of epithelium, subepithelial muscle layers, number of mast cells and goblet cells of group III were significantly ameliorated when compared with the group II. Although Hedera helix administration reduced only goblet cell counts and the thicknesses of basement membrane in the asthmatic airways, dexamethasone ameliorated all histopathologic parameters except thickness of basement membrane better than Hedera helix.

Modulating immunogenic properties of HIV-1 gp41 membrane-proximal external region by destabilizing six-helix bundle structure

Energy Technology Data Exchange (ETDEWEB)

Banerjee, Saikat; Shi, Heliang; Habte, Habtom H.; Qin, Yali; Cho, Michael W., E-mail: mcho@iastate.edu

2016-03-15

The C-terminal alpha-helix of gp41 membrane-proximal external region (MPER; {sup 671}NWFDITNWLWYIK{sup 683}) encompassing 4E10/10E8 epitopes is an attractive target for HIV-1 vaccine development. We previously reported that gp41-HR1-54Q, a trimeric protein comprised of the MPER in the context of a stable six-helix bundle (6HB), induced strong immune responses against the helix, but antibodies were directed primarily against the non-neutralizing face of the helix. To better target 4E10/10E8 epitopes, we generated four putative fusion intermediates by introducing double point mutations or deletions in the heptad repeat region 1 (HR1) that destabilize 6HB in varying degrees. One variant, HR1-∆10-54K, elicited antibodies in rabbits that targeted W672, I675 and L679, which are critical for 4E10/10E8 recognition. Overall, the results demonstrated that altering structural parameters of 6HB can influence immunogenic properties of the MPER and antibody targeting. Further exploration of this strategy could allow development of immunogens that could lead to induction of 4E10/10E8-like antibodies. - Highlights: • Four gp41 MPER-based immunogens that resemble fusion intermediates were generated. • C-terminal region of MPER that contains 4E10/10E8 epitopes was highly immunogenic. • Altering 6HB structure can influence immunogenic properties of the MPER. • Induced antibodies targeted multiple residues critical for 4E10/10E8 binding. • Development of immunogens based on fusion intermediates is a promising strategy.

Metabolism of [15N]alanine in the ectomycorrhizal fungus Paxillus involutus

International Nuclear Information System (INIS)

Chalot, M.; Finlay, R.D.; Ek, H.; Söderström, B.

1995-01-01

Chalot, M., Finlay, R. D., Ek, H., and Söderström, B. 1995. Metabolism of [ 15 N]alanine in the ectomycorrhizal fungus Paxillus involutus. Experimental Mycology 19, 297-304. Alanine metabolism in the ectomycorrhizal fungus Paxillus involutus was investigated using [ 15 N]alanine. Short-term exposure of mycelial discs to [ 15 N]alanine showed that the greatest flow of 15 N was to glutamate and to aspartate. Levels of enrichment were as high as 15-20% for glutamate and 13-18% for aspartate, whereas that of alanine reached 30%. Label was also detected in the amino-N of glutamine and in serine and glycine, although at lower levels. Preincubation of mycelia with aminooxyacetate, an inhibitor of transamination reactions. resulted in complete inhibition of the flow of the label to glutamate, aspartate, and amino-N of glutamine, whereas [ 15 N]alanine rapidly accumulated. This evidence indicates the direct involvement of alanine aminotransferase for translocation of 15 N from alanine to glutamate. Alanine may be a convenient reservoir of both nitrogen and carbon. (author)

The Alpha-Helix Concept: Innovative utilization of the Space Station Program. A report to the National Aeronautical and Space Administration requesting establishment of a Sensory Physiology Laboratory on the Space Station

Science.gov (United States)

Bandurski, R. S.; Singh, N.

1983-01-01

A major laboratory dedicated to biological-medical research is proposed for the Space Platform. The laboratory would focus on sensor physiology and biochemistry since sensory physiology represents the first impact of the new space environment on living organisms. Microgravity and the high radiation environment of space would be used to help solve the problems of prolonged sojourns in space but, more importantly, to help solve terrestrial problems of human health and agricultural productivity. The emphasis would be on experimental use of microorganisms and small plants and small animals to minimize the space and time required to use the Space Platform for maximum human betterment. The Alpha Helix Concept, that is, the use of the Space Platform to bring experimental biomedicine to a new and extreme frontier is introduced so as to better understand the worldly environment. Staffing and instrumenting the Space Platform biomedical laboratory in a manner patterned after successful terrestrial sensory physiology laboratories is also proposed.

Dimerization of the docking/adaptor protein HEF1 via a carboxy-terminal helix-loop-helix domain.

Science.gov (United States)

Law, S F; Zhang, Y Z; Fashena, S J; Toby, G; Estojak, J; Golemis, E A

1999-10-10

HEF1, p130(Cas), and Efs define a family of multidomain docking proteins which plays a central coordinating role for tyrosine-kinase-based signaling related to cell adhesion. HEF1 function has been specifically implicated in signaling pathways important for cell adhesion and differentiation in lymphoid and epithelial cells. While the SH3 domains and SH2-binding site domains (substrate domains) of HEF1 family proteins are well characterized and binding partners known, to date the highly conserved carboxy-terminal domains of the three proteins have lacked functional definition. In this study, we have determined that the carboxy-terminal domain of HEF1 contains a divergent helix-loop-helix (HLH) motif. This motif mediates HEF1 homodimerization and HEF1 heterodimerization with a recognition specificity similar to that of the transcriptional regulatory HLH proteins Id2, E12, and E47. We had previously demonstrated that the HEF1 carboxy-terminus expressed as a separate domain in yeast reprograms cell division patterns, inducing constitutive pseudohyphal growth. Here we show that pseudohyphal induction by HEF1 requires an intact HLH, further supporting the idea that this motif has an effector activity for HEF1, and implying that HEF1 pseudohyphal activity derives in part from interactions with yeast helix-loop-helix proteins. These combined results provide initial insight into the mode of function of the HEF1 carboxy-terminal domain and suggest that the HEF1 protein may interact with cellular proteins which control differentiation. Copyright 1999 Academic Press.

Use of alpha-amanitin as a transcriptional blocking agent in mouse embryos: a cautionary note

International Nuclear Information System (INIS)

Kidder, G.M.; Green, A.F.; McLachlin, J.R.

1985-01-01

We have tested the effect of alpha-amanitin at 10, 50 and 100 micrograms/ml, on precursor uptake and incorporation into poly(A)+ RNA and poly(A)- RNA of mouse embryos on days 2, 3 and 4 of gestation. Embryos were pretreated with the inhibitor for 2 hr, then labeled for 2 hr in its continued presence. RNA fractions were separated by affinity chromatography on oligo(dT)-cellulose. alpha-Amanitin did not suppress uptake of RNA precursors at any of the concentrations tested in any stage. At 10 micrograms/ml, we could not detect any effect on incorporation into either RNA fraction in any stage. Only the highest concentration tested, 100 micrograms/ml, was effective in all stages in substantially suppressing incorporation into poly(A)+ RNA within 2 hr. Longer treatments increased the level of suppression to a maximum of about 80%. Incorporation into poly(A)- RNA was suppressed to roughly the same extent. Despite previously reported data, it cannot be assumed that alpha-amanitin at concentrations less than 100 micrograms/ml brings about a quick interruption of mRNA synthesis in preimplantation mouse embryos

Performance effects of acute β-alanine induced paresthesia in competitive cyclists.

Science.gov (United States)

Bellinger, Phillip M; Minahan, Clare L

2016-01-01

β-alanine is a common ingredient in supplements consumed by athletes. Indeed, athletes may believe that the β-alanine induced paresthesia, experienced shortly after ingestion, is associated with its ergogenic effect despite no scientific mechanism supporting this notion. The present study examined changes in cycling performance under conditions of β-alanine induced paresthesia. Eight competitive cyclists (VO2max = 61.8 ± 4.2 mL·kg·min(-1)) performed three practices, one baseline and four experimental trials. The experimental trials comprised a 1-km cycling time trial under four conditions with varying information (i.e., athlete informed β-alanine or placebo) and supplement content (athlete received β-alanine or placebo) delivered to the cyclist: informed β-alanine/received β-alanine, informed placebo/received β-alanine, informed β-alanine/received placebo and informed placebo/received placebo. Questionnaires were undertaken exploring the cyclists' experience of the effects of the experimental conditions. A possibly likely increase in mean power was associated with conditions in which β-alanine was administered (±95% CL: 2.2% ± 4.0%), but these results were inconclusive for performance enhancement (p = 0.32, effect size = 0.18, smallest worthwhile change = 56% beneficial). A possibly harmful effect was observed when cyclists were correctly informed that they had ingested a placebo (-1.0% ± 1.9%). Questionnaire data suggested that β-alanine ingestion resulted in evident sensory side effects and six cyclists reported placebo effects. Acute ingestion of β-alanine is not associated with improved 1-km TT performance in competitive cyclists. These findings are in contrast to the athlete's "belief" as cyclists reported improved energy and the ability to sustain a higher power output under conditions of β-alanine induced paresthesia.

Alanine - Valine dynamics in pregnant rabbits | Emudianughe ...

African Journals Online (AJOL)

[15N]-alanine and [15N]–valine dynamics were studied in 29 -30 days pregnant New-Zealand rabbits. Over the experimental period, there was no detectable significant difference of mean ± SD of alanine concentrations within the sampling intervals in maternal, umbilical venous and arterial blood samples suggesting that ...

Isobaric PVT Behavior of Poly(Carbonate) (PC)

Science.gov (United States)

Grassia, Luigi; D'Amore, Alberto

2010-06-01

The scaling law for relaxation times, τ(V,T) = ℑ(TVγ), recently proposed by Casalini and Roland, is utilized in the framework of KAHR (Kovacs Aklonis Hutchinson and Ramos) phenomenological theory. With this approach it is shown that the Pressure, Volume, Temperature (PVT) data obtained on Poly(carbonate)(PC) can be reliably predicted, in the region of the alpha relaxation, by using only two fitting parameters, namely: the relaxation time in the reference condition, τg, and the fractional exponent, β that describes the dispersion of the alpha relaxation.

Peptide design using alpha,beta-dehydro amino acids: from beta-turns to helical hairpins.

Science.gov (United States)

Mathur, Puniti; Ramakumar, S; Chauhan, V S

2004-01-01

Incorporation of alpha,beta-dehydrophenylalanine (DeltaPhe) residue in peptides induces folded conformations: beta-turns in short peptides and 3(10)-helices in larger ones. A few exceptions-namely, alpha-helix or flat beta-bend ribbon structures-have also been reported in a few cases. The most favorable conformation of DeltaPhe residues are (phi,psi) approximately (-60 degrees, -30 degrees ), (-60 degrees, 150 degrees ), (80 degrees, 0 degrees ) or their enantiomers. DeltaPhe is an achiral and planar residue. These features have been exploited in designing DeltaPhe zippers and helix-turn-helix motifs. DeltaPhe can be incorporated in both right and left-handed helices. In fact, consecutive occurrence of three or more DeltaPhe amino acids induce left-handed screw sense in peptides containing L-amino acids. Weak interactions involving the DeltaPhe residue play an important role in molecular association. The C--H.O==C hydrogen bond between the DeltaPhe side-chain and backbone carboxyl moiety, pi-pi stacking interactions between DeltaPhe side chains belonging to enantiomeric helices have shown to stabilize folding. The unusual capability of a DeltaPhe ring to form the hub of multicentered interactions namely, a donor in aromatic C--H.pi and C--H.O==C and an acceptor in a CH(3).pi interaction suggests its exploitation in introducing long-range interactions in the folding of supersecondary structures. Copyright 2004 Wiley Periodicals, Inc. Biopolymers (Pept Sci), 2004

The basic helix-loop-helix region of the transcriptional repressor hairy and enhancer of split 1 is preorganized to bind DNA

NARCIS (Netherlands)

Popovic, Matija; Wienk, Hans; Coglievina, Maristella; Boelens, Rolf; Pongor, Sándor; Pintar, Alessandro

2014-01-01

Hairy and enhancer of split 1, one of the main downstream effectors in Notch signaling, is a transcriptional repressor of the basic helix-loop-helix (bHLH) family. Using nuclear magnetic resonance methods, we have determined the structure and dynamics of a recombinant protein, H1H, which includes an

IAEA reference dosimeter: Alanine-ESR

International Nuclear Information System (INIS)

Mehta, K.; Girzikowsky, R.

1999-01-01

Since 1985, the IAEA has been using alanine-ESR as a transfer dosimeter for its dose quality audit service, namely the International Dose Assurance Service. The alanine dosimeters are rod-type containing 70 wt% DL--α-alanine and 30 wt% polystyrene. We have two self-shielded gamma facilities for the calibration of the dosimetry system, where the temperature within the irradiation chamber can be controlled by a specially designed unit. A 4th order polynomial is fitted to the 16 data points in the dose range of 100 Gy to 50 kGy. The measured value of the irradiation temperature coefficient at two dose values (15 and 45 kGy) is 0.23 %/deg. C. Also, the ESR-response was followed for several dosimeters for about 8 months to study the post-irradiation effect. A value of 0.008 %/day was observed for the fading of the response for two dose values (15 and 45 kGy) and three irradiation temperatures (15, 27 and 40 deg. C). The effect of the analysis temperature on the ESR response was also studied. The combined relative uncertainty for the IAEA alanine-ESR dosimetry system is 1.5% (k=1). This includes that transferred from the primary laboratory for the dose rate measurements of the gamma facilities, dosimetry system calibration uncertainties, batch variability and uncertainty in the curve fitting procedure. This value however does not include the contribution due to the irradiation temperature correction which is applied when it differs from that during calibration; this component being specific for each dose measurement. (author)

Mode conversions by a discontinuous junction of two helix loaded waveguides

International Nuclear Information System (INIS)

Choe, J.Y.; Ahn, S.; Ganquly, A.K.; Uhm, H.S.

1983-01-01

For various reasons, it is desirable to vary the primary propagating mode from one section of the waveguide to another. We choose the base structure to be the sheath helix loaded waveguide. Specifically, we join two physically different helix loaded waveguides axisymmetrically, thereby providing the required discontinuities at the junction (Z = 0). The helix loaded waveguide is more advantageous to the simple waveguide in that the helix mode that exists uniquely in the helix waveguide in addition to the usual fast wave hybrid modes, is without cutoff and thus behaves like a transmission line. In order to obtain the mode conversion rates, we expand the waves in the both sides of the junction with its own eigenmodes including the evanescent modes, and by matching fields at the junction (Z = 0) obtain the matrix equation for the coefficients for the eigenmodes in both sides. By choosing the propagating incident wave (Z = 0) the resulting outgoing waves in the other end (Z > 0) will be computed from the matrix equation. A computer program is devised to solve the suitably truncated matrix equation, and the numerical examples for the mode conversion rates with the parameter variations will be presented. The relevant physical parameters to yield discontinuities at the junction are the radii of the outer conductor and the helix wire and the pitch angle of the helix. Special emphases are on the conversion rates from the helix mode (Z 0) for the application to the tapered gyrotron amplifier

Supramolecular Architectures and Mimics of Complex Natural Folds Derived from Rationally Designed alpha-Helical Protein Structures

Science.gov (United States)

Tavenor, Nathan Albert

Protein-based supramolecular polymers (SMPs) are a class of biomaterials which draw inspiration from and expand upon the many examples of complex protein quaternary structures observed in nature: collagen, microtubules, viral capsids, etc. Designing synthetic supramolecular protein scaffolds both increases our understanding of natural superstructures and allows for the creation of novel materials. Similar to small-molecule SMPs, protein-based SMPs form due to self-assembly driven by intermolecular interactions between monomers, and monomer structure determines the properties of the overall material. Using protein-based monomers takes advantage of the self-assembly and highly specific molecular recognition properties encodable in polypeptide sequences to rationally design SMP architectures. The central hypothesis underlying our work is that alpha-helical coiled coils, a well-studied protein quaternary folding motif, are well-suited to SMP design through the addition of synthetic linkers at solvent-exposed sites. Through small changes in the structures of the cross-links and/or peptide sequence, we have been able to control both the nanoscale organization and the macroscopic properties of the SMPs. Changes to the linker and hydrophobic core of the peptide can be used to control polymer rigidity, stability, and dimensionality. The gaps in knowledge that this thesis sought to fill on this project were 1) the relationship between the molecular structure of the cross-linked polypeptides and the macroscopic properties of the SMPs and 2) a means of creating materials exhibiting multi-dimensional net or framework topologies. Separate from the above efforts on supramolecular architectures was work on improving backbone modification strategies for an alpha-helix in the context of a complex protein tertiary fold. Earlier work in our lab had successfully incorporated unnatural building blocks into every major secondary structure (beta-sheet, alpha-helix, loops and beta

Characterization of the l-alanine exporter AlaE of Escherichia coli and its potential role in protecting cells from a toxic-level accumulation of l-alanine and its derivatives.

Science.gov (United States)

Kim, Seryoung; Ihara, Kohei; Katsube, Satoshi; Hori, Hatsuhiro; Ando, Tasuke; Isogai, Emiko; Yoneyama, Hiroshi

2015-08-01

We previously reported that the alaE gene of Escherichia coli encodes the l-alanine exporter AlaE. The objective of this study was to elucidate the mechanism of the AlaE exporter. The minimum inhibitory concentration of l-alanine and l-alanyl-l-alanine in alaE-deficient l-alanine-nonmetabolizing cells MLA301ΔalaE was 4- and >4000-fold lower, respectively, than in the alaE-positive parent cells MLA301, suggesting that AlaE functions as an efflux pump to avoid a toxic-level accumulation of intracellular l-alanine and its derivatives. Furthermore, the growth of the alaE-deficient mutant derived from the l-alanine-metabolizing strain was strongly inhibited in the presence of a physiological level of l-alanyl-l-alanine. Intact MLA301ΔalaE and MLA301ΔalaE/pAlaE cells producing plasmid-borne AlaE, accumulated approximately 200% and 50%, respectively, of the [(3) H]l-alanine detected in MLA301 cells, suggesting that AlaE exports l-alanine. When 200 mmol/L l-alanine-loaded inverted membrane vesicles prepared from MLA301ΔalaE/pAlaE were placed in a solution containing 200 mmol/L or 0.34 μmol/L l-alanine, energy-dependent [(3) H]l-alanine accumulation occurred under either condition. This energy-dependent uphill accumulation of [(3) H]l-alanine was strongly inhibited in the presence of carbonyl cyanide m-chlorophenylhydrazone but not by dicyclohexylcarbodiimide, suggesting that the AlaE-mediated l-alanine extrusion was driven by proton motive force. Based on these results, physiological roles of the l-alanine exporter are discussed. © 2015 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

EPR/alanine dosimetry for two therapeutic proton beams

Energy Technology Data Exchange (ETDEWEB)

Marrale, Maurizio, E-mail: maurizio.marrale@unipa.it [Dipartimento di Fisica e Chimica, Università di Palermo, Viale delle Scienze, Edificio 18, 90128 Palermo (Italy); Gruppo V Sezione INFN di Catania, Via Santa Sofia, 64, 95123 Catania (Italy); Carlino, Antonio [Dipartimento di Fisica e Chimica, Università di Palermo, Viale delle Scienze, Edificio 18, 90128 Palermo (Italy); EBG MedAustron GmbH, Marie Curie-Straße 5, A-2700 Wiener Neustadt (Austria); Gallo, Salvatore [Dipartimento di Fisica e Chimica, Università di Palermo, Viale delle Scienze, Edificio 18, 90128 Palermo (Italy); Gruppo V Sezione INFN di Catania, Via Santa Sofia, 64, 95123 Catania (Italy); Laboratorio PH3DRA, Dipartimento di Fisica e Astronomia, Università di Catania, Via Santa Sofia 64, 95123 Catania (Italy); Longo, Anna; Panzeca, Salvatore [Dipartimento di Fisica e Chimica, Università di Palermo, Viale delle Scienze, Edificio 18, 90128 Palermo (Italy); Gruppo V Sezione INFN di Catania, Via Santa Sofia, 64, 95123 Catania (Italy); Bolsi, Alessandra; Hrbacek, Jan; Lomax, Tony [Center for Proton Therapy, Paul Scherrer Institute, 5232 Villigen PSI (Switzerland)

2016-02-01

In this work the analysis of the electron paramagnetic resonance (EPR) response of alanine pellets exposed to two different clinical proton beams employed for radiotherapy is performed. One beam is characterized by a passive delivery technique and is dedicated to the eyes treatment (OPTIS2 beam line). Alanine pellets were irradiated with a 70 MeV proton beam corresponding to 35 mm range in eye tissue. We investigated how collimators with different sizes and shape used to conform the dose to the planned target volume influence the delivered dose. For this purpose we performed measurements with varying the collimator size (Output Factor) and the results were compared with those obtained with other dosimetric techniques (such as Markus chamber and diode detector). This analysis showed that the dosimeter response is independent of collimator diameter if this is larger than or equal to 10 mm. The other beam is characterized by an active spot-scanning technique, the Gantry1 beam line (maximum energy 230 MeV), and is used to treat deep-seated tumors. The dose linearity of alanine response in the clinical dose range was tested and the alanine dose response at selected locations in depth was measured and compared with the TPS planned dose in a quasi-clinical scenario. The alanine response was found to be linear in the dose in the clinical explored range (from 10 to 70 Gy). Furthermore, a depth dose profile in a quasi-clinical scenario was measured and compared to the dose computed by the Treatment Planning System PSIPLAN. The comparison of calibrated proton alanine measurements and TPS dose shows a difference under 1% in the SOBP and a “quenching” effect up to 4% in the distal part of SOBP. The positive dosimetric characteristics of the alanine pellets confirm the feasibility to use these detectors for “in vivo” dosimetry in clinical proton beams.

EPR/alanine dosimetry for two therapeutic proton beams

International Nuclear Information System (INIS)

Marrale, Maurizio; Carlino, Antonio; Gallo, Salvatore; Longo, Anna; Panzeca, Salvatore; Bolsi, Alessandra; Hrbacek, Jan; Lomax, Tony

2016-01-01

In this work the analysis of the electron paramagnetic resonance (EPR) response of alanine pellets exposed to two different clinical proton beams employed for radiotherapy is performed. One beam is characterized by a passive delivery technique and is dedicated to the eyes treatment (OPTIS2 beam line). Alanine pellets were irradiated with a 70 MeV proton beam corresponding to 35 mm range in eye tissue. We investigated how collimators with different sizes and shape used to conform the dose to the planned target volume influence the delivered dose. For this purpose we performed measurements with varying the collimator size (Output Factor) and the results were compared with those obtained with other dosimetric techniques (such as Markus chamber and diode detector). This analysis showed that the dosimeter response is independent of collimator diameter if this is larger than or equal to 10 mm. The other beam is characterized by an active spot-scanning technique, the Gantry1 beam line (maximum energy 230 MeV), and is used to treat deep-seated tumors. The dose linearity of alanine response in the clinical dose range was tested and the alanine dose response at selected locations in depth was measured and compared with the TPS planned dose in a quasi-clinical scenario. The alanine response was found to be linear in the dose in the clinical explored range (from 10 to 70 Gy). Furthermore, a depth dose profile in a quasi-clinical scenario was measured and compared to the dose computed by the Treatment Planning System PSIPLAN. The comparison of calibrated proton alanine measurements and TPS dose shows a difference under 1% in the SOBP and a “quenching” effect up to 4% in the distal part of SOBP. The positive dosimetric characteristics of the alanine pellets confirm the feasibility to use these detectors for “in vivo” dosimetry in clinical proton beams.

Alanine repeats influence protein localization in splicing speckles and paraspeckles.

Science.gov (United States)

Chang, Shuo-Hsiu; Chang, Wei-Lun; Lu, Chia-Chen; Tarn, Woan-Yuh

2014-12-16

Mammalian splicing regulatory protein RNA-binding motif protein 4 (RBM4) has an alanine repeat-containing C-terminal domain (CAD) that confers both nuclear- and splicing speckle-targeting activities. Alanine-repeat expansion has pathological potential. Here we show that the alanine-repeat tracts influence the subnuclear targeting properties of the RBM4 CAD in cultured human cells. Notably, truncation of the alanine tracts redistributed a portion of RBM4 to paraspeckles. The alanine-deficient CAD was sufficient for paraspeckle targeting. On the other hand, alanine-repeat expansion reduced the mobility of RBM4 and impaired its splicing activity. We further took advantage of the putative coactivator activator (CoAA)-RBM4 conjoined splicing factor, CoAZ, to investigate the function of the CAD in subnuclear targeting. Transiently expressed CoAZ formed discrete nuclear foci that emerged and subsequently separated-fully or partially-from paraspeckles. Alanine-repeat expansion appeared to prevent CoAZ separation from paraspeckles, resulting in their complete colocalization. CoAZ foci were dynamic but, unlike paraspeckles, were resistant to RNase treatment. Our results indicate that the alanine-rich CAD, in conjunction with its conjoined RNA-binding domain(s), differentially influences the subnuclear localization and biogenesis of RBM4 and CoAZ. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

The result of Alanine/ESR dosimetry at Wolsung unit 1

International Nuclear Information System (INIS)

Park, Byeong Ryong; Choi, Hoon; Lim, Young Khi

2008-01-01

It needs accurate estimation of radiation level for verifying machinery and cable in Nuclear Power Plant. Therefore, in this study, we used ESR(Electron Spin Resonance) system for estimate dose of Alanine dosimeter. Alanine/ESR dosimetry, already known as a dosimetric method in medical and industrial field, was applied to estimate dose quantity at cable locations within a nuclear power plant as a part of equipment qualification program. Alanine/ESR dosimetry of absorbed dose range is 1 - 100 KGy. The alanine dosimeter is not significantly affected by temperature and fading is limited to 1% per year. The alanine dosimeters were fixed on the targeted cable or nearest position to measure dose quantity to get accurate value. Alanine dosimeters were scanned by commercially used two different ESR systems, e-scan and EMX series for alanine dosimeters. To estimate more accurate dose, two environmental correction factors, irradiation temperature and dosimeter weight, were used in calculation of absorbed dose quantity. In this study, dose values which are alinine dosimeter from Wolsong unit 1 are measured by two ESR systems. And then the results was compared each other

Impact of charged amino acid substitution in the transmembrane domain of L-alanine exporter, AlaE, of Escherichia coli on the L-alanine export.

Science.gov (United States)

Kim, Seryoung; Ihara, Kohei; Katsube, Satoshi; Ando, Tasuke; Isogai, Emiko; Yoneyama, Hiroshi

2017-01-01

The Escherichia coli alaE gene encodes the L-alanine exporter, AlaE, that catalyzes active export of L-alanine using proton electrochemical potential. The transporter comprises only 149 amino acid residues and four predicted transmembrane domains (TMs), which contain three charged amino acid residues. The AlaE-deficient L-alanine non-metabolizing cells (ΔalaE cells) appeared hypersusceptible to L-alanyl-L-alanine showing a minimum inhibitory concentration (MIC) of 2.5 µg/ml for the dipeptide due to a toxic accumulation of L-alanine. To elucidate the mechanism by which AlaE exports L-alanine, we replaced charged amino acid residues in the TMs, glutamic acid-30 (TM-I), arginine-45 (TM-II), and aspartic acid-84 (TM-III) with their respective charge-conserved amino acid or a net neutral cysteine. The ΔalaE cells producing R45K or R45C appeared hypersusceptible to the dipeptide, indicating that arginine-45 is essential for AlaE activity. MIC of the dipeptide in the ΔalaE cells expressing E30D and E30C was 156 µg/ml and >10,000 µg/ml, respectively, thereby suggesting that a negative charge at this position is not essential. The ΔalaE cells expressing D84E or D84C showed an MIC >10,000 and 78 µg/ml, respectively, implying that a negative charge is required at this position. These results were generally consistent with that of the L-alanine accumulation experiments in intact cells. We therefore concluded that charged amino acid residues (R45 and D84) in the AlaE transmembrane domain play a pivotal role in L-alanine export. Replacement of three cysteine residues at C22, C28 (both in TM-I), and C135 (C-terminal region) with alanine showed only a marginal effect on L-alanine export.

L-alanine as a precursor of ethylamine in camellia sinensis

International Nuclear Information System (INIS)

Takeo, Tadakazu

1975-01-01

After absorption of ammonium nitrogen, nitrogen-deficient Camellia sinensis synthesized theanine following synthesis of glutamic acid and alanine. The rate of incorporation of 14 C from L-alanine U- 14 C into theanine was faster than from acetaldehyde 1-2 14 C. Incorporation of 14 C from L-alanine U- 14 C into the ethylamide of theanine was prevented by adding an excess of ethylamine to the culture solution. Green seedlings converted alanine to ethylamine more rapidly than did etiolated seedlings. (auth.)

Mechanical unfolding reveals stable 3-helix intermediates in talin and α-catenin.

Directory of Open Access Journals (Sweden)

Vasyl V Mykuliak

2018-04-01

Full Text Available Mechanical stability is a key feature in the regulation of structural scaffolding proteins and their functions. Despite the abundance of α-helical structures among the human proteome and their undisputed importance in health and disease, the fundamental principles of their behavior under mechanical load are poorly understood. Talin and α-catenin are two key molecules in focal adhesions and adherens junctions, respectively. In this study, we used a combination of atomistic steered molecular dynamics (SMD simulations, polyprotein engineering, and single-molecule atomic force microscopy (smAFM to investigate unfolding of these proteins. SMD simulations revealed that talin rod α-helix bundles as well as α-catenin α-helix domains unfold through stable 3-helix intermediates. While the 5-helix bundles were found to be mechanically stable, a second stable conformation corresponding to the 3-helix state was revealed. Mechanically weaker 4-helix bundles easily unfolded into a stable 3-helix conformation. The results of smAFM experiments were in agreement with the findings of the computational simulations. The disulfide clamp mutants, designed to protect the stable state, support the 3-helix intermediate model in both experimental and computational setups. As a result, multiple discrete unfolding intermediate states in the talin and α-catenin unfolding pathway were discovered. Better understanding of the mechanical unfolding mechanism of α-helix proteins is a key step towards comprehensive models describing the mechanoregulation of proteins.

Analysis of an Alanine/Arginine Mixture by Using TLC/FTIR Technique

Directory of Open Access Journals (Sweden)

Jun Liu

2014-01-01

Full Text Available We applied TLC/FTIR coupled with mapping technique to analyze an alanine/arginine mixture. Narrow band TLC plates prepared by using AgI as a stationary phase were used to separate alanine and arginine. The distribution of alanine and arginine spots was manifested by a 3D chromatogram. Alanine and arginine can be successfully separated by the narrow band TLC plate. In addition, the FTIR spectra of the separated alanine and arginine spots on the narrow band TLC plate are roughly the same as the corresponding reference IR spectra.

EPR of alanine irradiated by neutrons

International Nuclear Information System (INIS)

Pivovarov, S.P.; Seredavina, T.A.; Zhdanov, S.V.; Mul'gin, S.I.; Zhakparov, R.K.

2001-01-01

In the work the first results of EPR studies of alanine, irradiated with diverse doses at neutron cyclotron generator different conditions and on the critical reactor stand are presented. A dose linearity dependence of EPR signal is observing, the methods of γ-background contribution separation are discussed. Obtain results is giving the basis to recommendation of alanine as an effective detector irradiation. However it is demanded the farther study on clarification of radiation sensitivity value dependence on the neutron energy spectrum form

Conductivity of alanine solution for high level dosimetry

International Nuclear Information System (INIS)

Wieser, A.; Figel, M.; Regulla, D.F.

1993-01-01

The amino acid alanine is well known as a dosimetric detector material for high level dosimetry. Its application is based on the formation of radicals by ionising radiation. The free radicals are earlier detected by electron spin resonance (ESR) spectroscopy or chemically after dissolving the irradiated samples. Of all these methods the ESR/alanine system is the most advanced and is suggested for reference dosimetry. At present, however, the high cost of the system is a serious handicap for a large scale routine application in radiation plants. In this study the variation of electrical conductivity of L-alanine solution with applied dose is investigated in the range from 0.5-200 kGy. The conductivity was measured with a 50 MHz RF oscillator. This readout method is uncomplicated and may be suitable for routine application. The experiments were performed with L-alanine solution in glass ampoules. (Author)

DNA-like double helix formed by peptide nucleic acid

DEFF Research Database (Denmark)

Wittung, P; Nielsen, Peter E.; Buchardt, O

1994-01-01

Although the importance of the nucleobases in the DNA double helix is well understood, the evolutionary significance of the deoxyribose phosphate backbone and the contribution of this chemical entity to the overall helical structure and stability of the double helix is not so clear. Peptide nucleic...

Alanine EPR dosimeter response in proton therapy beams

International Nuclear Information System (INIS)

Gall, K.; Serago, C.; Desrosiers, M.; Bensen, D.

1997-01-01

We report a series of measurements directed to assess the suitability of alanine as a mailable dosimeter for dosimetry quality assurance of proton radiation therapy beams. These measurements include dose-response of alanine at 140 MeV, and comparison of response vs energy with a parallel plate ionization chamber. All irradiations were made at the Harvard Cyclotron Laboratory, and the dosimeters were read at NIST. The results encourage us that alanine could be expected to serve as a mailable dosimeter with systematic error due to differential energy response no greater than 3% when doses of 25 Gy are used. (Author)

Hydroxyproline Ring Pucker Causes Frustration of Helix Parameters in the Collagen Triple Helix

Science.gov (United States)

Ying Chow, W.; Bihan, Dominique; Forman, Chris J.; Slatter, David A.; Reid, David G.; Wales, David J.; Farndale, Richard W.; Duer, Melinda J.

2015-07-01

Collagens, the most abundant proteins in mammals, are defined by their triple-helical structures and distinctive Gly-Xaa-Yaa repeating sequence, where Xaa is often proline and Yaa, hydroxyproline (Hyp/O). It is known that hydroxyproline in the Yaa position stabilises the triple helix, and that lack of proline hydroxylation in vivo leads to dysfunctional collagen extracellular matrix assembly, due to a range of factors such as a change in hydration properties. In addition, we note that in model peptides, when Yaa is unmodified proline, the Xaa proline has a strong propensity to adopt an endo ring conformation, whilst when Yaa is hydroxyproline, the Xaa proline adopts a range of endo and exo conformations. Here we use a combination of solid-state NMR spectroscopy and potential energy landscape modelling of synthetic triple-helical collagen peptides to understand this effect. We show that hydroxylation of the Yaa proline causes the Xaa proline ring conformation to become metastable, which in turn confers flexibility on the triple helix.

Complexation of vanadium (v) with alanine in different ionic strength

International Nuclear Information System (INIS)

Garib, F.; Zare, K.; Fekri, H

2002-01-01

The formation constants of species formed in the system H ++ alanine and VO 2 + alanine have be determined in aqueous solution for 1.0 3 NaCIO 4 ,using a combination of pramiracetam and spectrophotometric techniques. The compositions of the formed complexes and their stability constants were determined ny curve fitting method and it was shown that di oxovanadium(V) forms two mononuclear 1:1 and 1:2 species with alanine of the type VO 2 L and VO 2 L 2 The protonation constant of the amino group of alanine has been determined using a computer program which employ a least-squares method. The defence of the protonation of alanine and the stability constant of the species on ionic strength are described by a Debby-huckel type equation

Alanine/ESR dosimetry system for routine use in radiation processing

International Nuclear Information System (INIS)

Kojima, T.; Haruyama, Y.; Tachibana, H.; Tanaka, R.; Okamoto, J.; Hara, H.; Kashiwazaki, S.

1993-01-01

A new alanine-polystyrene(PS) dosimeter prepared with simplified molding procedure and an automatic desk-top dose-reader of alanine dosimeter were developed for the purpose of routine use. Combination of these two allows us to apply a reliable alanine/ESR dosimetry system to routine dosimetric process control in industrial gamma radiation processing. (Author)

Nature of the Charged-Group Effect on the Stability of the C-Peptide Helix

Science.gov (United States)

Shoemaker, Kevin R.; Kim, Peter S.; Brems, David N.; Marqusee, Susan; York, Eunice J.; Chaiken, Irwin M.; Stewart, John M.; Baldwin, Robert L.

1985-04-01

The residues responsible for the pH-dependent stability of the helix formed by the isolated C-peptide (residues 1-13 of ribonuclease A) have been identified by chemical synthesis of analogues and measurement of their helix-forming properties. Each of the residues ionizing between pH 2 and pH 8 has been replaced separately by an uncharged residue. Protonation of Glu-2- is responsible for the sharp decrease in helix stability between pH 5 and pH 2, and deprotonation of His-12+ causes a similar decrease between pH 5 and pH 8. Glu-9- is not needed for helix stability. The results cannot be explained by the Zimm-Bragg model and host-guest data for α -helix formation, which predict that the stability of the C-peptide helix should increase when Glu-2- is protonated or when His-12+ is deprotonated. Moreover, histidine+ is a strong helix-breaker in host-guest studies. In proteins, acidic and basic residues tend to occur at opposite ends of α -helices: acidic residues occur preferentially near the NH2-terminal end and basic residues near the COOH-terminal end. A possible explanation, based on a helix dipole model, has been given [Blagdon, D. E. & Goodman, M. (1975) Biopolymers 14, 241-245]. Our results are consistent with the helix dipole model and they support the suggestion that the distribution of charged residues in protein helices reflects the helix-stabilizing propensity of those residues. Because Glu-9 is not needed for helix stability, a possible Glu-9-\\cdots His-12+ salt bridge does not contribute significantly to helix stability. The role of a possible Glu-2-\\cdots Arg-10+ salt bridge has not yet been evaluated. A charged-group effect on α -helix stability in water has also been observed in a different peptide system [Ihara, S., Ooi, T. & Takahashi, S. (1982) Biopolymers 21, 131-145]: block copolymers containing (Ala)20 and (Glu)20 show partial helix formation at low temperatures, pH 7.5, where the glutamic acid residues are ionized. (Glu)20(Ala)20Phe forms a

Emergence of the persistent spin helix in semiconductor quantum wells

International Nuclear Information System (INIS)

Koralek, Jake; Weber, Chris; Orenstein, Joe; Bernevig, Andrei; Zhang, Shoucheng; Mack, Shawn; Awschalom, David

2008-01-01

According to Noether's theorem, for every symmetry in nature there is a corresponding conservation law. For example, invariance with respect to spatial translation corresponds to conservation of momentum. In another well-known example, invariance with respect to rotation of the electron's spin, or SU(2) symmetry, leads to conservation of spin polarization. For electrons in a solid, this symmetry is ordinarily broken by spin-orbit (SO) coupling, allowing spin angular momentum to flow to orbital angular momentum. However, it has recently been predicted that SU(2) can be recovered in a two-dimensional electron gas (2DEG), despite the presence of SO coupling. The corresponding conserved quantities include the amplitude and phase of a helical spin density wave termed the 'persistent spin helix' (PSH) .2 SU(2) is restored, in principle, when the strength of two dominant SO interactions, the Rashba (alpha) and linear Dresselhaus (beta 1), are equal. This symmetry is predicted to be robust against all forms of spin-independent scattering, including electron-electron interactions, but is broken by the cubic Dresselhaus term (beta 3) and spin-dependent scattering. When these terms are negligible, the distance over which spin information can propagate is predicted to diverge as alpha approaches beta 1. Here we observe experimentally the emergence of the PSH in GaAs quantum wells (QW's) by independently tuning alpha and beta 1. Using transient spin-grating spectroscopy (TSG), we find a spin-lifetime enhancement of two orders of magnitude near the symmetry point. Excellent quantitative agreement with theory across a wide range of sample parameters allows us to obtain an absolute measure of all relevant SO terms, identifying beta 3 as the main SU(2) violating term in our samples. The tunable suppression of spin-relaxation demonstrated in this work is well-suited for application to spintronics

Reproducibility and signal response linearity of Alanine gel dosimeter

International Nuclear Information System (INIS)

Silva, Cleber Feijo Silva; Campos, Leticia Lucente

2008-01-01

Gel Dosimetry has been studied mainly for medical applications, because it presents signal response in the dose range used in radiotherapy treatments and it can be applied for three dimensional dosimetry. Alanine gel dosimeter is a new gel material developed at IPEN that presents significant improvement on previous alanine systems developed by Costa (1994). The DL-Alanine (C 3 H 7 NO 2 ) is an amino acid tissue equivalent that improves the production of ferric ions in the solution. These ferric ions concentration can be measured by spectrophotometry technique. This work aims to study the reproducibility of the alanine gel solutions and the signal response as a function of gamma radiation dose, considering that these two properties are very important for characterizing and standardizing any dosimeter. (author)

Polyphosphate-dependent synthesis of ATP and ADP by the family-2 polyphosphate kinases in bacteria.

Science.gov (United States)

Nocek, Boguslaw; Kochinyan, Samvel; Proudfoot, Michael; Brown, Greg; Evdokimova, Elena; Osipiuk, Jerzy; Edwards, Aled M; Savchenko, Alexei; Joachimiak, Andrzej; Yakunin, Alexander F

2008-11-18

Inorganic polyphosphate (polyP) is a linear polymer of tens or hundreds of phosphate residues linked by high-energy bonds. It is found in all organisms and has been proposed to serve as an energy source in a pre-ATP world. This ubiquitous and abundant biopolymer plays numerous and vital roles in metabolism and regulation in prokaryotes and eukaryotes, but the underlying molecular mechanisms for most activities of polyP remain unknown. In prokaryotes, the synthesis and utilization of polyP are catalyzed by 2 families of polyP kinases, PPK1 and PPK2, and polyphosphatases. Here, we present structural and functional characterization of the PPK2 family. Proteins with a single PPK2 domain catalyze polyP-dependent phosphorylation of ADP to ATP, whereas proteins containing 2 fused PPK2 domains phosphorylate AMP to ADP. Crystal structures of 2 representative proteins, SMc02148 from Sinorhizobium meliloti and PA3455 from Pseudomonas aeruginosa, revealed a 3-layer alpha/beta/alpha sandwich fold with an alpha-helical lid similar to the structures of microbial thymidylate kinases, suggesting that these proteins share a common evolutionary origin and catalytic mechanism. Alanine replacement mutagenesis identified 9 conserved residues, which are required for activity and include the residues from both Walker A and B motifs and the lid. Thus, the PPK2s represent a molecular mechanism, which potentially allow bacteria to use polyP as an intracellular energy reserve for the generation of ATP and survival.

High Energy Electron Dosimetry by Alanine/ESR Spectroscopy

International Nuclear Information System (INIS)

Chu, Sung Sil

1989-01-01

Dosimetry based on electron spin resonance(ESR) analysis of radiation induced free radicals in amino acids is relevant to biological dosimetry applications. Alanine detectors are without walls and are tissue equivalent. Therefore, alanine ESR dosimetry looks promising for use in the therapy level. The dose range of the alanine/ESR dosimetry system can be extended down to l Gy. In a water phantom the absorbed dose of electrons generated by a medical linear accelerator of different initial energies (6-21 MeV) and therapeutic dose levels(1-60 Gy) was measured. Furthermore, depth dose measurements carried out with alanine dosimeters were compared with ionization chamber measurements. As the results, the measured absorbed doses for shallow depth of initial electron energies above 15 MeV were higher by 2-5% than those calculated by nominal energy CE factors. This seems to be caused by low energy scattered beams generated from the scattering foil and electron cones of beam projecting device in medical linear accelerator

Alanine Enhances Aminoglycosides-Induced ROS Production as Revealed by Proteomic Analysis

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Jin-zhou Ye

2018-01-01

Full Text Available Metabolite-enabled killing of antibiotic-resistant pathogens by antibiotics is an attractive strategy to manage antibiotic resistance. Our previous study demonstrated that alanine or/and glucose increased the killing efficacy of kanamycin on antibiotic-resistant bacteria, whose action is through up-regulating TCA cycle, increasing proton motive force and enhancing antibiotic uptake. Despite the fact that alanine altered several metabolic pathways, other mechanisms could be potentially involved in alanine-mediated kanamycin killing of bacteria which remains to be explored. In the present study, we adopted proteomic approach to analyze the proteome changes induced by exogenous alanine. Our results revealed that the expression of three outer membrane proteins was altered and the deletion of nagE and fadL decreased the intracellular kanamycin concentration, implying their possible roles in mediating kanamycin transport. More importantly, the integrated analysis of proteomic and metabolomic data pointed out that alanine metabolism could connect to riboflavin metabolism that provides the source for reactive oxygen species (ROS production. Functional studies confirmed that alanine treatment together with kanamycin could promote ROS production that in turn potentiates the killing of antibiotic-resistant bacteria. Further investigation showed that alanine repressed the transcription of antioxidant-encoding genes, and alanine metabolism to riboflavin metabolism connected with riboflavin metabolism through TCA cycle, glucogenesis pathway and pentose phosphate pathway. Our results suggest a novel mechanism by which alanine facilitates kanamycin killing of antibiotic-resistant bacteria via promoting ROS production.

Higher Ratio of Serum Alpha-Fetoprotein Could Predict Outcomes in Patients with Hepatitis B Virus-Associated Hepatocellular Carcinoma and Normal Alanine Aminotransferase.

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Young-Il Kim

Full Text Available The role of serum alpha-fetoprotein (AFP levels in the surveillance and diagnosis of hepatocellular carcinoma (HCC is controversial. The aim of this study was to investigate the value of serially measured serum AFP levels in HCC progression or recurrence after initial treatment.A total of 722 consecutive patients newly diagnosed with HCC and treated at the National Cancer Center, Korea, between January 2004 and December 2009 were enrolled. The AFP ratios between 4-8 weeks post-treatment and those at the time of HCC progression or recurrence were obtained. Multivariate logistic regression analysis was performed to correlate the post-treatment AFP ratios with the presence of HCC progression or recurrence.The etiology of HCC was related to chronic hepatitis B virus (HBV infection in 562 patients (77.8%, chronic hepatitis C virus (HCV infection in 74 (10.2%, and non-viral cause in 86 (11.9%. There was a significant decrease in serum AFP levels from the baseline to 4 to 8 weeks after treatment (median AFP, 319.6 ng/mL vs. 49.6 ng/mL; p 1.0 was an independently associated with HCC progression or recurrence. Among the different causes of HCC analyzed, this association was significant only for HCC related to chronic hepatitis B (p< 0.001 and non-viral causes (p<0.05, and limited only to patients who had normal alanine aminotransferase (ALT levels.Serial measurements of serum AFP ratios could be helpful in detecting progression or recurrence in treated patients with HBV-HCC and normal ALT.

Higher Ratio of Serum Alpha-Fetoprotein Could Predict Outcomes in Patients with Hepatitis B Virus-Associated Hepatocellular Carcinoma and Normal Alanine Aminotransferase

Science.gov (United States)

Park, Joong-Won

2016-01-01

Background The role of serum alpha-fetoprotein (AFP) levels in the surveillance and diagnosis of hepatocellular carcinoma (HCC) is controversial. The aim of this study was to investigate the value of serially measured serum AFP levels in HCC progression or recurrence after initial treatment. Methods A total of 722 consecutive patients newly diagnosed with HCC and treated at the National Cancer Center, Korea, between January 2004 and December 2009 were enrolled. The AFP ratios between 4–8 weeks post-treatment and those at the time of HCC progression or recurrence were obtained. Multivariate logistic regression analysis was performed to correlate the post-treatment AFP ratios with the presence of HCC progression or recurrence. Results The etiology of HCC was related to chronic hepatitis B virus (HBV) infection in 562 patients (77.8%), chronic hepatitis C virus (HCV) infection in 74 (10.2%), and non-viral cause in 86 (11.9%). There was a significant decrease in serum AFP levels from the baseline to 4 to 8 weeks after treatment (median AFP, 319.6 ng/mL vs. 49.6 ng/mL; p 1.0 was an independently associated with HCC progression or recurrence. Among the different causes of HCC analyzed, this association was significant only for HCC related to chronic hepatitis B (p< 0.001) and non-viral causes (p<0.05), and limited only to patients who had normal alanine aminotransferase (ALT) levels. Conclusion Serial measurements of serum AFP ratios could be helpful in detecting progression or recurrence in treated patients with HBV-HCC and normal ALT. PMID:27304617

"Special Issue": Regional Dimensions of the Triple Helix Model

Science.gov (United States)

Todeva, Emanuela; Danson, Mike

2016-01-01

This paper introduces the rationale for the special issue and its contributions, which bridge the literature on regional development and the Triple Helix model. The concept of the Triple Helix at the sub-national, and specifically regional, level is established and examined, with special regard to regional economic development founded on…

A smallest 6 kda metalloprotease, mini-matrilysin, in living world: a revolutionary conserved zinc-dependent proteolytic domain- helix-loop-helix catalytic zinc binding domain (ZBD

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Yu Wei-Hsuan

2012-05-01

Full Text Available Abstract Background The Aim of this study is to study the minimum zinc dependent metalloprotease catalytic folding motif, helix B Met loop-helix C, with proteolytic catalytic activities in metzincin super family. The metzincin super family share a catalytic domain consisting of a twisted five-stranded β sheet and three long α helices (A, B and C. The catalytic zinc is at the bottom of the cleft and is ligated by three His residues in the consensus sequence motif, HEXXHXXGXXH, which is located in helix B and part of the adjacent Met turn region. An interesting question is - what is the minimum portion of the enzyme that still possesses catalytic and inhibitor recognition?” Methods We have expressed a 60-residue truncated form of matrilysin which retains only the helix B-Met turn-helix C region and deletes helix A and the five-stranded β sheet which form the upper portion of the active cleft. This is only 1/4 of the full catalytic domain. The E. coli derived 6 kDa MMP-7 ZBD fragments were purified and refolded. The proteolytic activities were analyzed by Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 peptide assay and CM-transferrin zymography analysis. SC44463, BB94 and Phosphoramidon were computationally docked into the 3day structure of the human MMP7 ZBD and TAD and thermolysin using the docking program GOLD. Results This minimal 6 kDa matrilysin has been refolded and shown to have proteolytic activity in the Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2 peptide assay. Triton X-100 and heparin are important factors in the refolding environment for this mini-enzyme matrilysin. This minienzyme has the proteolytic activity towards peptide substrate, but the hexamer and octamer of the mini MMP-7 complex demonstrates the CM-transferrin proteolytic activities in zymographic analysis. Peptide digestion is inhibited by SC44463, specific MMP7 inhibitors, but not phosphorimadon. Interestingly, the mini MMP-7 can be processed by autolysis and producing ~ 6�

Hybrid conducting polymer materials incorporating poly-oxo-metalates for extraction of actinides; Materiaux polymeres conducteurs hybrides incorporant des polyoxometallates pour l'extraction d'actinides

Energy Technology Data Exchange (ETDEWEB)

Racimor, D

2003-09-15

The preparation and characterization of hybrid conducting polymers incorporating poly-oxo-metalates for extracting actinides is discussed. A study of the coordination of various lanthanide cations (Ce(III), Ce(IV), Nd(III)) by the mono-vacant poly-oxo-metalate {alpha}{sub 2}-[P{sub 2}W{sub 17}O{sub 61}]{sup 10-} showed significant differences according to the cation.. Various {alpha}-A-[PW{sub 9}O{sub 34}(RPO){sub 2}]{sup 5-} hybrids were synthesized and their affinity for actinides or lanthanides was demonstrated through complexation. The first hybrid poly-oxo-metallic lanthanide complexes were then synthesized, as was the first hybrid functionalized with a pyrrole group. The electro-polymerization conditions of this pyrrole remain still to be optimized. Poly-pyrrole materials incorporating {alpha}{sub 2}-[P{sub 2}W{sub 17}O{sub 61}]{sup 10-} or its neodymium or cerium complexes as doping agents proved to be the first conducting polymer incorporating poly-oxo-metalates capable of extracting plutonium from nitric acid. (author)

Investigating the effect of a single glycine to alanine substitution on interactions of antimicrobial peptide latarcin 2a with a lipid membrane.

Science.gov (United States)

Idiong, Grace; Won, Amy; Ruscito, Annamaria; Leung, Bonnie O; Hitchcock, Adam P; Ianoul, Anatoli

2011-09-01

Latarcins are linear, α-helical antimicrobial peptides purified from the venom of the Central Asian spider Lachesana tarabaevi, with lytic activity against Gram-positive and Gram-negative bacteria, erythrocytes, and yeast at micromolar concentrations. In this work, we investigated the role of the hinge in latarcin 2a (ltc2a, GLFGKLIKKFGRKAISYAVKKARGKH-COOH), which adopts a helix-hinge-helix conformation in membrane-mimicking environments, on peptide-membrane interactions and its potential effect on the selective toxicity of the peptide. A modified latarcin 2a, ltc2aG11A, obtained by replacing the glycine at position 11 with alanine (ltc2aG11A, GLFGKLIKKFARKAISYAVKKARGKH-COOH), adopts a more rigid structure due to the reduced conformational flexibility. Langmuir monolayer measurements combined with atomic force microscopy and X-ray photoemission electron microscopy (X-PEEM) indicate that both peptides bind and insert preferentially into anionic compared with zwitterionic phospholipid monolayers. Modified ltc2aG11A was found to be more disruptive of supported phospholipid bilayer modeling mammalian cell membrane. However, no considerable difference in lytic activity of the two peptides toward bacterial membrane was found. Overall the data indicate that decrease in the flexibility of ltc2a induced by the modification in the hinge region is likely to increase the peptide's nonspecific interactions with zwitterionic cell membranes and potentially increase its toxicity against eukaryotic cells.

Efficient L-Alanine Production by a Thermo-Regulated Switch in Escherichia coli.

Science.gov (United States)

Zhou, Li; Deng, Can; Cui, Wen-Jing; Liu, Zhong-Mei; Zhou, Zhe-Min

2016-01-01

L-Alanine has important applications in food, pharmaceutical and veterinary and is used as a substrate for production of engineered thermoplastics. Microbial fermentation could reduce the production cost and promote the application of L-alanine. However, the presence of L-alanine significantly inhibit cell growth rate and cause a decrease in the ultimate L-alanine productivity. For efficient L-alanine production, a thermo-regulated genetic switch was designed to dynamically control the expression of L-alanine dehydrogenase (alaD) from Geobacillus stearothermophilus on the Escherichia coli B0016-060BC chromosome. The optimal cultivation conditions for the genetically switched alanine production using B0016-060BC were the following: an aerobic growth phase at 33 °C with a 1-h thermo-induction at 42 °C followed by an oxygen-limited phase at 42 °C. In a bioreactor experiment using the scaled-up conditions optimized in a shake flask, B0016-060BC accumulated 50.3 g biomass/100 g glucose during the aerobic growth phase and 96 g alanine/100 g glucose during the oxygen-limited phase, respectively. The L-alanine titer reached 120.8 g/l with higher overall and oxygen-limited volumetric productivities of 3.09 and 4.18 g/l h, respectively, using glucose as the sole carbon source. Efficient cell growth and L-alanine production were reached separately, by switching cultivation temperature. The results revealed the application of a thermo-regulated strategy for heterologous metabolic production and pointed to strategies for improving L-alanine production.

Corrosion inhibition of nickel in H2SO4 solution by alanine

International Nuclear Information System (INIS)

Hamed, E.; Abd El-REhim, S.S.; El-Shahat, M.F.; Shaltot, A.M.

2012-01-01

Highlights: ► Corrosion of Ni in 1 M H 2 SO 4 in the absence and the presence of alanine. ► Alanine acts as a moderate mixed type inhibitor. ► Physical adsorption of alanine and formation of protective film are on Ni surface. ► Addition of KI improves the inhibition efficiency (synergistic effect). ► EFM technique is in reasonably good agreement with the different techniques used. - Abstract: The effect of alanine, as a safe inhibitor, was studied by measuring the corrosion of Ni in aerated and stagnant 1 M H 2 SO 4 solution (pH ∼0.2). Measurements were performed under various conditions using potentiodynamic polarization, electrochemical impedance spectroscopy (EIS) and the new electrochemical frequency modulation (EFM) methods. The obtained results showed that the addition of alanine alone gives a moderate inhibition and acts as an anodic-type inhibitor. The inhibition is due to physical adsorption of alanine on the metal surface. The inhibition efficiency enhances with increasing alanine concentration and immersion time but decreases with rise in temperature. The apparent activation energy, E a , is higher in the presence than in the absence of alanine. Addition of I − ions greatly improves the inhibition efficiency of alanine. The synergistic effect is due to enhanced adsorption of alanine cations by chemisorbed I − anions on the metal surface. The results obtained from polarization, EIS and EFM techniques are in good agreement indicating that EFM method can be used successfully for monitoring corrosion rate of Ni in H 2 SO 4 solution with and without alanine.

Preparation of α-alanine-3H by the interaction of atomic tritium heated up to 2000 K with a solid alanine target at 77 K

International Nuclear Information System (INIS)

Filatov, Eh.S.; Simonov, E.F.; Shishkov, A.V.; Mogil'nikov, V.P.

1979-01-01

Absorption of hydrogen by alanine targets, the target behaviour and the yield of α-alanine- 3 H were studied in experiments involving straight passage of H and T atoms from the sourse (2000 K) to the target (77 K) as a function of the exposure time. In the studies with 3 H 2 the radioactivity of the gas phase was decreasing more rapidly than the overall pressure of hydrogen: H 3 H accumulates more rapidly in the gas phase. Alanine decomposition products were identified. The conditions for the studies of α-alanine- 3 H are suggested

Two alanine racemase genes in Salmonella typhimurium that differ in structure and function.

OpenAIRE

Wasserman, S A; Walsh, C T; Botstein, D

1983-01-01

Mutations were isolated in a previously undescribed Salmonella typhimurium gene encoding an alanine racemase essential for utilization of L-alanine as a source of carbon, energy, and nitrogen. This new locus, designated dadB, lies within one kilobase of the D-alanine dehydrogenase locus (dadA), which is also required for alanine catabolism. The dadA and dadB genes are coregulated. Mutants (including insertions) lacking the dadB alanine racemase do not require D-alanine for growth unless a mut...

EPR of gamma-irradiated polycrystalline alanine-in-glass dosimeter

International Nuclear Information System (INIS)

Al-Karmi, Anan M.; Morsy, M.A.

2008-01-01

This study attempts to overcome some of the reported discrepancies in alanine-EPR reproducibility that may be related to alanine dosimeter preparation and/or EPR spectrometer settings. The dosimeters were prepared by packing pure polycrystalline L-α-alanine directly as supplied by the manufacturer in glass tubes. This dosimeter production scheme avoids any possible contribution to the EPR signal from a binding material. The dosimeters were irradiated with gamma ray to low-dose ranges typical for medical therapy (0-20 Gy). Special attention has been paid to the study of minimum detectable dose, measurement repeatability and reproducibility, and post-irradiation stability. The dosimeter exhibited a linear dose response in the dose range from 0.1 to 20 Gy. These positive properties favor the polycrystalline alanine-in-glass tube as a radiation dosimeter

Beta-alanine and dopamine in the reddish brown scales of Papilio butterflies

International Nuclear Information System (INIS)

Umebachi, Yoshishige; Ishizaki, Yumi

1983-01-01

(1) Reddish brown scales of the anal eye spot in the hind-wings of P. demoleus and P. machaon have been examined for β-alanine and dopamine. (2) The scales were fractionated into 70% ethanol-soluble fraction, 4% HCl-methanol-soluble fraction, and the residua l scales, and the β-alanine content of each fraction was determined. Most of the β-alanine present in the scales has been found in the residual scales. On acid hydrolysis of the residual scales, the β-alanine has been rather rapidly released, and the hydrolysate has contained a large amount of β-alanine. (3) The protein-bound brown pigment (HCl-ppt fraction), which was extracted with 1 N NaOH and precipitated by being acidified with HCl, has contained a large amount of β-alanine. In most or at least some of the β-alanine, the NH 2 -group has been proved to be free. (4) 14 C-Labelled β-alanine and 14 C-dopamine, which were injected at prepupal or pupal stage, have been incorporated in the highest degree into the residual scales. And the 14 C has been confirmed to be present in the HCl-ppt fraction. (5) All these results indicate that the pigment of the reddish brown scales contains β-alanine and dopamine. (author)

Evolutionary Diversification of Alanine Transaminases in Yeast: Catabolic Specialization and Biosynthetic Redundancy

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Ximena Escalera-Fanjul

2017-06-01

Full Text Available Gene duplication is one of the major evolutionary mechanisms providing raw material for the generation of genes with new or modified functions. The yeast Saccharomyces cerevisiae originated after an allopolyploidization event, which involved mating between two different ancestral yeast species. ScALT1 and ScALT2 codify proteins with 65% identity, which were proposed to be paralogous alanine transaminases. Further analysis of their physiological role showed that while ScALT1 encodes an alanine transaminase which constitutes the main pathway for alanine biosynthesis and the sole pathway for alanine catabolism, ScAlt2 does not display alanine transaminase activity and is not involved in alanine metabolism. Moreover, phylogenetic studies have suggested that ScALT1 and ScALT2 come from each one of the two parental strains which gave rise to the ancestral hybrid. The present work has been aimed to the understanding of the properties of the ancestral type Lacchancea kluyveri LkALT1 and Kluyveromyces lactis KlALT1, alanine transaminases in order to better understand the ScALT1 and ScALT2 evolutionary history. These ancestral -type species were chosen since they harbor ALT1 genes, which are related to ScALT2. Presented results show that, although LkALT1 and KlALT1 constitute ScALT1 orthologous genes, encoding alanine transaminases, both yeasts display LkAlt1 and KlAlt1 independent alanine transaminase activity and additional unidentified alanine biosynthetic and catabolic pathway(s. Furthermore, phenotypic analysis of null mutants uncovered the fact that KlAlt1 and LkAlt1 have an additional role, not related to alanine metabolism but is necessary to achieve wild type growth rate. Our study shows that the ancestral alanine transaminase function has been retained by the ScALT1 encoded enzyme, which has specialized its catabolic character, while losing the alanine independent role observed in the ancestral type enzymes. The fact that ScAlt2 conserves 64

Alanine-ESR dosimeter: application for dosimetry in industrial electron beam accelerator

International Nuclear Information System (INIS)

Murali, S.; Venkataramani, R.; Pushparaja; Sarma, K.S.S.; Natarajan, V.; Sastry, M.D.

2000-01-01

The feasibility of DL-α-alanine, as ESR dosimeter in powder form, was examined under the conditions of pulse electron accelerator used as an industrial irradiator. The investigations were carried out to examine the following aspects: (i) Alanine-ESR dose response in irradiator characteristics viz. various beam energies, beam currents, product conveying speeds, (ii) linearity of dose response of irradiated alanine signal for suitable range, (iii) dose uniformity of the irradiated samples and (iv) depth dose measurements using alanine powder dosimeters sandwiched between polyethylene layers. Experiments were carried out by varying some of the irradiator parameters at mobile mode of the conveyor (product under movement) and also at stationary mode for different EB energies and pulse rates. For estimation of EB dose, signal intensities of gamma irradiated DL--alanine powder calibrated with Fricke dosimetry have been used. Feasibility of application of alanine ESR dosimeter for low dose measurement down to 350 Gy has been experimentally established. The present studies show that under variable operating conditions of irradiator, alanine ESR dosimetry is suitable for dosimetric applications from low dose (350 Gy) to high dose (53 kgy). (author)

A molecular receptor selective for zwitterionic alanine.

Science.gov (United States)

Rubio, Omayra H; Taouil, Rachid; Muñiz, Francisco M; Monleón, Laura M; Simón, Luis; Sanz, Francisca; Morán, Joaquín R

2017-01-04

A molecular receptor has been synthesized joining an aza-crown ether with a chiral chromane which mimics the oxyanion hole of the enzymes. With this receptor an apolar host-guest complex with zwitterionic alanine has been achieved through the formation of up to seven H-bonds. This complex allows the extraction of aqueous alanine to a chloroform phase, while other natural amino acids are poorly extracted or are not extracted at all. Due to the chiral nature of the receptor, enantioselective extraction from the aqueous alanine solution to a chloroform phase takes place. X-Ray analysis combined with anisotropic effects, NOE and CD studies revealed the absolute configuration of both strong and weak complexes. Modelling studies also support the proposed structures. The presence of an oxyanion-hole motif in this structure was corroborated by X-ray diffraction studies.

Compound-specific nitrogen isotope analysis of D-alanine, L-alanine, and valine: application of diastereomer separation to delta15N and microbial peptidoglycan studies.

Science.gov (United States)

Takano, Yoshinori; Chikaraishi, Yoshito; Ogawa, Nanako O; Kitazato, Hiroshi; Ohkouchi, Naohiko

2009-01-01

We have developed an analytical method to determine the compound-specific nitrogen isotope compositions of individual amino acid enantiomers using gas chromatography/combustion/isotope ratio mass spectrometry. A novel derivatization of amino acid diastereomers by optically active (R)-(-)-2-butanol or (S)-(+)-2-butanol offers two advantages for nitrogen isotope analysis. First, chromatographic chiral separation can be achieved without the use of chiral stationary-phase columns. Second, the elution order of these compounds on the chromatogram can be switched by a designated esterification reaction. We applied the method to the compound-specific nitrogen isotope analysis of D- and L-alanine in a peptidoglycan derived from the cell walls of cultured bacteria (Firmicutes and Actinobacteria; Enterococcus faecalis, Staphylococcus aureus, Staphylococcus staphylolyticus, Lactobacillus acidophilus, Bacillus subtilis, Micrococcus luteus, and Streptomyces sp.), natural whole bacterial cells (Bacillus subtilis var. natto), (pseudo)-peptidoglycan from archaea (Methanobacterium sp.), and cell wall from eukaryota (Saccharomyces cerevisiae). We observed statistically significant differences in nitrogen isotopic compositions; e.g., delta15N ( per thousand vs air) in Staphylococcus staphylolyticus for d-alanine (19.2 +/- 0.5 per thousand, n = 4) and L-alanine (21.3 +/- 0.8 per thousand, n = 4) and in Bacillus subtilis for D-alanine (6.2 +/- 0.2 per thousand, n = 3) and L-alanine (8.2 +/- 0.4 per thousand, n = 3). These results suggest that enzymatic reaction pathways, including the alanine racemase reaction, produce a nitrogen isotopic difference in amino acid enantiomers, resulting in 15N-depleted D-alanine. This method is expected to facilitate compound-specific nitrogen isotope studies of amino acid stereoisomers.

The C-Terminal RpoN Domain of sigma54 Forms an unpredictedHelix-Turn-Helix Motif Similar to domains of sigma70

Energy Technology Data Exchange (ETDEWEB)

Doucleff, Michaeleen; Malak, Lawrence T.; Pelton, Jeffrey G.; Wemmer, David E.

2005-11-01

The ''{delta}'' subunit of prokaryotic RNA-polymerase allows gene-specific transcription initiation. Two {sigma} families have been identified, {sigma}{sup 70} and {sigma}{sup 54}, which use distinct mechanisms to initiate transcription and share no detectable sequence homology. Although the {sigma}{sup 70}-type factors have been well characterized structurally by x-ray crystallography, no high-resolution structural information is available for the {sigma}{sup 54}-type factors. Here we present the NMR derived structure of the C-terminal domain of {sigma}{sup 54} from Aquifex aeolicus. This domain (Thr323 to Gly389), which contains the highly conserved RpoN box sequence, consists of a poorly structured N-terminal tail followed by a three-helix bundle, which is surprisingly similar to domains of the {sigma}{sup 70}-type proteins. Residues of the RpoN box, which have previously been shown to be critical for DNA binding, form the second helix of an unpredicted helix-turn-helix motif. This structure's homology with other DNA binding proteins, combined with previous biochemical data, suggest how the C-terminal domain of {sigma}{sup 54} binds to DNA.

Dosimetry of electron and gamma radiation with DL-alanine

International Nuclear Information System (INIS)

Costa, Z.M. da; Campos, L.L.

1996-01-01

A dosimetric method based on the quantitative determination of stabilised free radicals in irradiated crystalline DL-alanine by electron spin resonance (ESR) spectroscopy was proposed as early in 1962. Since then, alanine dosemeters owing to their unique properties have been investigated by many authors and used in dosimetry of various types of radiation, namely gamma rays, electron and neutrons. Alanine is a simple aminoacid, on irradiation at room temperature predominantly free paramagnetic radicals of the type CH 3 -CH-COOH are produced. This paper reports the application of powder DL-alanine/ESR dosemeter for measurement of absorbed dose of gamma radiation from 60 Co sources and reactor nucleus and electron beams from accelerator. The obtained results give useful information about the instrumental care necessary to obtain the needed overall accuracy in determination of absorbed dose. (author)

Deletions in the fifth alpha helix of HIV-1 matrix block virus release

International Nuclear Information System (INIS)

Sanford, Bridget; Li, Yan; Maly, Connor J.; Madson, Christian J.; Chen, Han; Zhou, You; Belshan, Michael

2014-01-01

The matrix (MA) protein of HIV-1 is the N-terminal component of the Gag structural protein and is critical for the early and late stages of viral replication. MA contains five α-helices (α1–α5). Deletions in the N-terminus of α5 as small as three amino acids impaired virus release. Electron microscopy of one deletion mutant (MA∆96-120) showed that its particles were tethered to the surface of cells by membranous stalks. Immunoblots indicated all mutants were processed completely, but mutants with large deletions had alternative processing intermediates. Consistent with the EM data, MA∆96-120 retained membrane association and multimerization capability. Co-expression of this mutant inhibited wild type particle release. Alanine scanning mutation in this region did not affect virus release, although the progeny virions were poorly infectious. Combined, these data demonstrate that structural ablation of the α5 of MA inhibits virus release. - Highlights: • Deletions were identified in the C-terminus of matrix that block virus release. • These deletion mutants still multimerized and associated with membranes. • TEM showed the mutant particles were tethered to the cell surface. • Amino acid mutagenesis of the region did not affect release. • The data suggests that disruption of matrix structure blocks virus release

Deletions in the fifth alpha helix of HIV-1 matrix block virus release

Energy Technology Data Exchange (ETDEWEB)

Sanford, Bridget; Li, Yan; Maly, Connor J.; Madson, Christian J. [Department of Medical Microbiology and Immunology, Creighton University, 2500 California Plaza, Omaha, NE 68178 (United States); Chen, Han [Center for Biotechnology, University of Nebraska-Lincoln, Lincoln, NE (United States); Zhou, You [Center for Biotechnology, University of Nebraska-Lincoln, Lincoln, NE (United States); Nebraska Center for Virology, Lincoln, NE (United States); Belshan, Michael, E-mail: michaelbelshan@creighton.edu [Department of Medical Microbiology and Immunology, Creighton University, 2500 California Plaza, Omaha, NE 68178 (United States); Nebraska Center for Virology, Lincoln, NE (United States)

2014-11-15

The matrix (MA) protein of HIV-1 is the N-terminal component of the Gag structural protein and is critical for the early and late stages of viral replication. MA contains five α-helices (α1–α5). Deletions in the N-terminus of α5 as small as three amino acids impaired virus release. Electron microscopy of one deletion mutant (MA∆96-120) showed that its particles were tethered to the surface of cells by membranous stalks. Immunoblots indicated all mutants were processed completely, but mutants with large deletions had alternative processing intermediates. Consistent with the EM data, MA∆96-120 retained membrane association and multimerization capability. Co-expression of this mutant inhibited wild type particle release. Alanine scanning mutation in this region did not affect virus release, although the progeny virions were poorly infectious. Combined, these data demonstrate that structural ablation of the α5 of MA inhibits virus release. - Highlights: • Deletions were identified in the C-terminus of matrix that block virus release. • These deletion mutants still multimerized and associated with membranes. • TEM showed the mutant particles were tethered to the cell surface. • Amino acid mutagenesis of the region did not affect release. • The data suggests that disruption of matrix structure blocks virus release.

Synthesis and GGCT Inhibitory Activity of N-Glutaryl-L-alanine Analogues.

Science.gov (United States)

Ii, Hiromi; Yoshiki, Tatsuhiro; Hoshiya, Naoyuki; Uenishi, Jun'ichi

2016-01-01

γ-Glutamylcyclotransferase (GGCT) is an important enzyme that cleaves γ-glutamyl-amino acid in the γ-glutamyl cycle to release 5-oxoproline and amino acid. Eighteen N-acyl-L-alanine analogues including eleven new compounds have been synthesized and examined for their inhibitory activity against recombinant human GGCT protein. Simple N-glutaryl-L-alanine was found to be the most potent inhibitor for GGCT. Other N-glutaryl-L-alanine analogues having methyl and dimethyl substituents at the 2-position were moderately effective, while N-(3R-aminoglutary)-L-alanine, the substrate having an (R)-amino group at the 3-position or N-(N-methyl-3-azaglutaryl)-L-alanine, the substrate having an N-methyl substituent on the 3-azaglutaryl carbon, in constract, exhibited excellent inhibition properties.

Stereoselective aminoacylation of a dinucleoside monophosphate by the imidazolides of DL-alanine and N-(tert-butoxycarbonyl)-DL-alanine

Science.gov (United States)

Profy, A. T.; Usher, D. A.

1984-01-01

The aminoacylation of diinosine monophosphate was studied experimentally. When the acylating agent was the imidazolide of N-(tert-butoxycarbonyl)-DL-alanine, a 40 percent enantiomeric excess of the isomer was incorporated at the 2' site and the positions of equilibrium for the reversible 2'-3' migration reaction differed for the D and L enantiomers. The reactivity of the nucleoside hydroxyl groups was found to decrease on the order 2'(3') less than internal 2' and less than 5', and the extent of the reaction was affected by the concentration of the imidazole buffer. Reaction of IpI with imidazolide of unprotected DL-alanine, by contrast, led to an excess of the D isomer at the internal 2' site. Finally, reaction with the N-carboxy anhydride of DL-alanine occurred without stereoselection. These results are found to be relevant to the study of the evolution of optical chemical activity and the origin of genetically directed protein synthesis.

Identification of basic/helix-loop-helix transcription factors reveals candidate genes involved in anthocyanin biosynthesis from the strawberry white-flesh mutant

OpenAIRE

Zhao, Fengli; Li, Gang; Hu, Panpan; Zhao, Xia; Li, Liangjie; Wei, Wei; Feng, Jiayue; Zhou, Houcheng

2018-01-01

As the second largest transcription factor family in plant, the basic helix-loop-helix (bHLH) transcription factor family, characterized by the conserved bHLH domain, plays a central regulatory role in many biological process. However, the bHLH transcription factor family of strawberry has not been systematically identified, especially for the anthocyanin biosynthesis. Here, we identified a total of 113 bHLH transcription factors and described their chromosomal distribution and bioinformatics...

Application of a Biodegradable Polyesteramide Derived from L-Alanine as Novel Excipient for Controlled Release Matrix Tablets.

Science.gov (United States)

Bonillo Martínez, Ana Dora; Galán, Inés Carmen Rodríguez; Bellver, María Victoria Margarit

2017-11-01

This pre-formulation study assays the capacity of the polyesteramide PADAS, poly (L-alanine-dodecanediol-L-alanine-sebacic), as an insoluble tablet excipient matrix for prolonged drug release. The flow properties of PADAS were suitable for tableting, and the compressibility of tablets containing exclusively PADAS was evaluated by ESEM observation of the microstructure. The tablets were resistant to crushing and non-friable and they did not undergo disintegration (typical features of an inert matrix). Tablets containing 33.33% sodium diclofenac (DF), ketoprofen (K) or dexketoprofen trometamol (DK-T) as a model drug, in addition with 66.67% of polymer, were formulated, and the absence of interactions between the components was confirmed by differential scanning calorimetry. Dissolution tests showed that PADAS retained DF and K and prolonged drug release, following a Higuchi kinetic. The tablets containing DK-T did not retain the drug sufficiently for prolonged release to be established. Tablets containing DK-T and 66.67, 83.33 or 91.67% PADAS, compressed at 44.48 or 88.96 kN, were elaborated to determine the influence of the polymer amount and of the compression force on DK-T release. Both parameters significantly delayed drug release, except when the proportion of polymer was 91.67%.

21 CFR 862.1030 - Alanine amino transferase (ALT/SGPT) test system.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Alanine amino transferase (ALT/SGPT) test system... Test Systems § 862.1030 Alanine amino transferase (ALT/SGPT) test system. (a) Identification. An alanine amino transferase (ALT/SGPT) test system is a device intended to measure the activity of the...

TOWARDS UNDERSTANDING OF HELIX B BASED CONFORMATIONAL DISEASES IN SERPIN

Directory of Open Access Journals (Sweden)

Mohamad Aman Jairajpuri

2012-12-01

Full Text Available Serine protease inhibitors (serpins are a unique family of protease inhibitors that are prone to polymer formation due to their metastable nature and a complex inhibition mechanism that involves large scale conformational change. Helix B is in the shutter region near the strand 2A and strand 3A of �-sheet A, where reactive centre loop inserts during the serpin inhibition mechanism. Helix B region in serpins is a mutation hotspot for naturally occurring variants that result in pathological conditions due to polymerization. Helix B residues are completely buried in the native state and loop inserted latent state but not in the inhibitory loop inserted cleaved conformation. Native to cleaved transition during inhibition forms a large cavity in the shutter region, which invariably is the largest cavity in most serpins in native state. In a recent paper we had for the first time hypothesized that exposure of helix B at the N-terminal end is important for smooth insertion of the reactive center loop during serpin inhibition mechanism. It is therefore possible that natural variant that induces conformational deformation of helix B probably alter the cavity size which increases the rate of loop-sheet interaction between the monomers resulting in increased polymerization.

In Quest of the Alanine R3 Radical: Multivariate EPR Spectral Analyses of X-Irradiated Alanine in the Solid State.

Science.gov (United States)

Jåstad, Eirik O; Torheim, Turid; Villeneuve, Kathleen M; Kvaal, Knut; Hole, Eli O; Sagstuen, Einar; Malinen, Eirik; Futsaether, Cecilia M

2017-09-28

The amino acid l-α-alanine is the most commonly used material for solid-state electron paramagnetic resonance (EPR) dosimetry, due to the formation of highly stable radicals upon irradiation, with yields proportional to the radiation dose. Two major alanine radical components designated R1 and R2 have previously been uniquely characterized from EPR and electron-nuclear double resonance (ENDOR) studies as well as from quantum chemical calculations. There is also convincing experimental evidence of a third minor radical component R3, and a tentative radical structure has been suggested, even though no well-defined spectral signature has been observed experimentally. In the present study, temperature dependent EPR spectra of X-ray irradiated polycrystalline alanine were analyzed using five multivariate methods in further attempts to understand the composite nature of the alanine dosimeter EPR spectrum. Principal component analysis (PCA), maximum likelihood common factor analysis (MLCFA), independent component analysis (ICA), self-modeling mixture analysis (SMA), and multivariate curve resolution (MCR) were used to extract pure radical spectra and their fractional contributions from the experimental EPR spectra. All methods yielded spectral estimates resembling the established R1 spectrum. Furthermore, SMA and MCR consistently predicted both the established R2 spectrum and the shape of the R3 spectrum. The predicted shape of the R3 spectrum corresponded well with the proposed tentative spectrum derived from spectrum simulations. Thus, results from two independent multivariate data analysis techniques strongly support the previous evidence that three radicals are indeed present in irradiated alanine samples.

40 CFR 721.520 - Alanine, N-(2-carboxyethyl)-N-alkyl-, salt.

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Alanine, N-(2-carboxyethyl)-N-alkyl... Specific Chemical Substances § 721.520 Alanine, N-(2-carboxyethyl)-N-alkyl-, salt. (a) Chemical substance... alanine, N-(2-carboxyethyl)-N- alkyl-, salt (P-89-336) is subject to reporting under this section for the...

cDNA cloning and characterization of Type I procollagen alpha1 chain in the skate Raja kenojei.

Science.gov (United States)

Hwang, Jae-Ho; Yokoyama, Yoshihiro; Mizuta, Shoshi; Yoshinaka, Reiji

2006-05-01

A full-length cDNA of the Type I procollagen alpha1 [pro-alpha1(I)] chain (4388 bp), coding for 1463 amino acid residues in the total length, was determined by RACE PCR using a cDNA library constructed from 4-week embryo of the skate Raja kenojei. The helical region of the skate pro-alpha1(I) chain consisted of 1014 amino acid residues - the same as other fibrillar collagen alpha chains from higher vertebrates. Comparison on denaturation temperatures of Type I collagens from the skate, rainbow trout (Oncorhynchus mykiss) and rat (Rattus norvegicus) revealed that the number of Gly-Pro-Pro and Gly-Gly in the alpha1(I) chains could be directly related to the thermal stability of the helix. The expression property of the skate pro-alpha1(I) chain mRNA and phylogenetic analysis with other vertebrate pro-alpha1(I) chains suggested that skate pro-alpha1(I) chain could be a precursor form of the skate Type I collagen alpha1 chain. The present study is the first evidence for the primary structure of full-length pro-alpha1(I) chain in an elasmobranch.

Nucleic acid helix structure determination from NMR proton chemical shifts

Energy Technology Data Exchange (ETDEWEB)

Werf, Ramon M. van der; Tessari, Marco; Wijmenga, Sybren S., E-mail: S.Wijmenga@science.ru.nl [Radboud University Nijmegen, Department of Biophysical Chemistry, Institute of Molecules and Materials (Netherlands)

2013-06-15

We present a method for de novo derivation of the three-dimensional helix structure of nucleic acids using non-exchangeable proton chemical shifts as sole source of experimental restraints. The method is called chemical shift de novo structure derivation protocol employing singular value decomposition (CHEOPS) and uses iterative singular value decomposition to optimize the structure in helix parameter space. The correct performance of CHEOPS and its range of application are established via an extensive set of structure derivations using either simulated or experimental chemical shifts as input. The simulated input data are used to assess in a defined manner the effect of errors or limitations in the input data on the derived structures. We find that the RNA helix parameters can be determined with high accuracy. We finally demonstrate via three deposited RNA structures that experimental proton chemical shifts suffice to derive RNA helix structures with high precision and accuracy. CHEOPS provides, subject to further development, new directions for high-resolution NMR structure determination of nucleic acids.

Control of alanine metabolism in rat liver by transport processes or cellular metabolism.

OpenAIRE

Fafournoux, P; Rémésy, C; Demigné, C

1983-01-01

1. Factors governing hepatic utilization of alanine were studied in vivo and in vitro in rats adapted to increasing dietary protein. 2. Hepatic alanine utilization was enhanced 5-fold with a 90%-casein diet, compared with a 13%-casein diet. The increased uptake resulted from enhanced fractional extraction in the presence of high concentrations of alanine in the portal vein. 3. The increase in alanine metabolism on high-protein diets was associated with an increase in alanine aminotransferase ...

Dose intercomparison study involving Fricke, ethanol chlorobenzene, PMMA and alanine dosimeters

International Nuclear Information System (INIS)

Lanuza, L.G.; Cabalfin, E.G.; Kojima, T.; Tachibana, H.

1999-01-01

A dose intercomparison study was carried out between the Philippine Nuclear Research Institute (PNRI) and Takasaki Radiation Chemistry Research Establishment, Japan Atomic Energy Research Institute (JAERI) to determine reliability of the dosimetry systems being used by PNRI employing ethanol chlorobenzene (ECB), Fricke and alanine dosimeters. The Fricke and ECB dosimeters were prepared at PNRI while the alanine-polystyrene dosimeter was provided by JAERI. Fricke or ECB dosimeters were irradiated together with alanine at PNRI gamma irradiation facilities. Analyses of the Fricke and ECB dosimeters were performed at PNRI while alanine dosimeters were analyzed at JAERI. A comparison study between alanine and polymethylmethacrylate (PMMA, Radix RN15) dosimeters was also undertaken at JAERI. The dosimeters were irradiated together under different irradiation conditions using the gamma irradiation facilities of JAERI and Radia Industry Co. Ltd. (Japan). Evaluations of PMMA and alanine dosimeters were both performed at JAERI. Result of the dose intercomparison of PNRI with the International Atomic Energy Agency through the International Dose Assurance Service (IDAS) is also presented. (author)

Dose intercomparison study involving Fricke, ethanol chlorobenzene, PMMA and alanine dosimeters

Energy Technology Data Exchange (ETDEWEB)

Lanuza, L G; Cabalfin, E G [Philippine Nuclear Research Institute, Quezon City (Philippines); Kojima, T; Tachibana, H [Takasaki Radiation Chemistry Research Establishment, Japan Atomic Energy Research institute, Takasaki (Japan)

1999-03-01

A dose intercomparison study was carried out between the Philippine Nuclear Research Institute (PNRI) and Takasaki Radiation Chemistry Research Establishment, Japan Atomic Energy Research Institute (JAERI) to determine reliability of the dosimetry systems being used by PNRI employing ethanol chlorobenzene (ECB), Fricke and alanine dosimeters. The Fricke and ECB dosimeters were prepared at PNRI while the alanine-polystyrene dosimeter was provided by JAERI. Fricke or ECB dosimeters were irradiated together with alanine at PNRI gamma irradiation facilities. Analyses of the Fricke and ECB dosimeters were performed at PNRI while alanine dosimeters were analyzed at JAERI. A comparison study between alanine and polymethylmethacrylate (PMMA, Radix RN15) dosimeters was also undertaken at JAERI. The dosimeters were irradiated together under different irradiation conditions using the gamma irradiation facilities of JAERI and Radia Industry Co. Ltd. (Japan). Evaluations of PMMA and alanine dosimeters were both performed at JAERI. Result of the dose intercomparison of PNRI with the International Atomic Energy Agency through the International Dose Assurance Service (IDAS) is also presented. (author) 8 refs, 3 figs, 4 tabs

The antiproton depth–dose curve measured with alanine detectors

CERN Document Server

Bassler, Niels; Palmans, Hugo; Holzscheiter, Michael H; Kovacevic, Sandra

2008-01-01

n this paper we report on the measurement of the antiproton depth–dose curve, with alanine detectors. The results are compared with simulations using the particle energy spectrum calculated by FLUKA, and using the track structure model of Hansen and Olsen for conversion of calculated dose into response. A good agreement is observed between the measured and calculated relative effectiveness although an underestimation of the measured values beyond the Bragg-peak remains unexplained. The model prediction of response of alanine towards heavy charged particles encourages future use of the alanine detectors for dosimetry of mixed radiation fields.

The Antiproton Depth Dose Curve Measured with Alanine Detectors

DEFF Research Database (Denmark)

Bassler, Niels; Hansen, Johnny Witterseh; Palmans, Hugo

2008-01-01

In this paper we report on the measurement of the antiproton depth dose curve, with alanine detectors. The results are compared with simulations using the particle energy spectrum calculated by FLUKA, and using the track structure model of Hansen et Olsen for conversion of calculated dose...... into response. A good agreement was observed between the measured and calculated relative effectiveness although a slight underestimation of the calculated values in the Bragg peak remains unexplained. The model prediction of response of alanine towards heavy charged particles encourages future use...... of the alanine detectors for dosimetry of mixed radiation fields....

A High Sensitivity EPR Technique for Alanine Dosimetry (invited paper)

International Nuclear Information System (INIS)

Haskell, E.H.; Hayes, R.B.; Kenner, G.H.

1998-01-01

Uncertainties of ± 5 mGy were achieved in the measurement of alanine dosemeters using optimised EPR parameters, instrumentation, spectral manipulation and subtraction techniques. Modulation amplitude and microwave power were adjusted to combine resonances of two neighbouring alanine signals. Instrumental variations were minimised by combining and subtracting pre- and post-measurement spectra of the empty EPR tube. A spectrum of the native signal of non-dosed alanine was generated from a single batch of dosemeters and subtracted from spectra of the irradiated dosemeters, also from the same batch. Field alignment was adjusted with the use of an in-cavity Mn ++ standard. A constant rotation goniometer was used to eliminate anisotropies in the EPR tube and alanine samples. Finally, digital filters were applied to the resulting spectra. (author)

Genome-wide identification of basic helix-loop-helix and NF-1 motifs underlying GR binding sites in male rat hippocampus

DEFF Research Database (Denmark)

Pooley, John R.; Flynn, Ben P.; Grøntved, Lars

2017-01-01

linked to structural and organizational roles, an absence of major tethering partners for GRs, and little or no evidence for binding at negative glucocorticoid response elements. A basic helix-loop-helix motif closely resembling a NeuroD1 or Olig2 binding site was found underlying a subset of GR binding......Glucocorticoids regulate hippocampal function in part by modulating gene expression through the glucocorticoid receptor (GR). GR binding is highly cell type specific, directed to accessible chromatin regions established during tissue differentiation. Distinct classes of GR binding sites...

Regional Dimensions of the Triple Helix Model: Setting the Context

Science.gov (United States)

Todeva, Emanuela; Danson, Mike

2016-01-01

This paper introduces the rationale for the special issue and its contributions, which bridge the literature on regional development and the Triple Helix model. The concept of the Triple Helix at the sub-national, and specifically regional, level is established and examined, with special regard to regional economic development founded on…

FMRFamide receptors of Helix aspersa

International Nuclear Information System (INIS)

Payza, K.

1988-01-01

A receptor binding assay and an isolated heart bioassay were used to identify and characterize the FMRFamide receptors in Helix. In the heart bioassay, FMRFamide increased myocardial contraction force. A potent FMRFamide analog, desaminoTyr-Phe-norLeu-arg-Phe-amide (daYFnLRFamide), was used as a radioiodinated receptor ligand. The high affinity binding of 125 I-daYFnLRFamide at 0 degree C to Helix brain membranes was reversible, saturable, pH-dependent and specific, with a K D of 13-14 nM. A lower affinity (245 nM) site was also observed. Radioligand binding sites were also identified in the heart, male reproductive organs and digestive organs. The structure-activity relations (SAR) of cardiostimulation correlated with the specificity of 125 I-daYFnLRFamide binding to brain and heart receptors. The SAR were similar to those of other molluscan FMRFamide bioassays, except that they showed a marked preference for some analogs with blocked amino-terminals

Elevated temperature triggers human respiratory syncytial virus F protein six-helix bundle formation

International Nuclear Information System (INIS)

Yunus, Abdul S.; Jackson, Trent P.; Crisafi, Katherine; Burimski, Irina; Kilgore, Nicole R.; Zoumplis, Dorian; Allaway, Graham P.; Wild, Carl T.; Salzwedel, Karl

2010-01-01

Human respiratory syncytial virus (RSV) is a major cause of severe lower respiratory tract infection in infants, immunocompromised patients, and the elderly. The RSV fusion (F) protein mediates fusion of the viral envelope with the target cell membrane during virus entry and is a primary target for antiviral drug and vaccine development. The F protein contains two heptad repeat regions, HR1 and HR2. Peptides corresponding to these regions form a six-helix bundle structure that is thought to play a critical role in membrane fusion. However, characterization of six-helix bundle formation in native RSV F protein has been hindered by the fact that a trigger for F protein conformational change has yet to be identified. Here we demonstrate that RSV F protein on the surface of infected cells undergoes a conformational change following exposure to elevated temperature, resulting in the formation of the six-helix bundle structure. We first generated and characterized six-helix bundle-specific antibodies raised against recombinant peptides modeling the RSV F protein six-helix bundle structure. We then used these antibodies as probes to monitor RSV F protein six-helix bundle formation in response to a diverse array of potential triggers of conformational changes. We found that exposure of 'membrane-anchored' RSV F protein to elevated temperature (45-55 deg. C) was sufficient to trigger six-helix bundle formation. Antibody binding to the six-helix bundle conformation was detected by both flow cytometry and cell-surface immunoprecipitation of the RSV F protein. None of the other treatments, including interaction with a number of potential receptors, resulted in significant binding by six-helix bundle-specific antibodies. We conclude that native, untriggered RSV F protein exists in a metastable state that can be converted in vitro to the more stable, fusogenic six-helix bundle conformation by an increase in thermal energy. These findings help to better define the mechanism of

Determination of the dose of traffic in HDR brachytherapy with ALANINE/R PE technique

International Nuclear Information System (INIS)

Guzman Calcina, C. S.; Chen, F.; Almeida, A. de; Baffa, O.

2001-01-01

It determines, experimentally, the dose of traffic in brachytherapy for High Dose Rate (HDR), using for the first-time the Electronic Paramagnetic Resonance (EPR) technique with alanine detectors. The value obtained is the published next to obtained using lithium fluoride thermoluminescent dosimeters [es

Importance of intrahepatic mechanisms to gluconeogenesis from alanine during exercise and recovery

International Nuclear Information System (INIS)

Wasserman, D.H.; Williams, P.E.; Lacy, D.B.; Green, D.R.; Cherrington, A.D.

1988-01-01

These studies were performed to assess the importance of intrahepatic mechanisms to gluconeogenesis in the dog during 150 min of treadmill exercise and 90 min of recovery. Sampling catheters were implanted in an artery and portal and hepatic veins 16 days before experimentation. Infusions of [U- 14 C]alanine, [3- 3 H]glucose, and indocyanine green were used to assess gluconeogenesis. During exercise, a decline in arterial and portal vein plasma alanine and in hepatic blood flow led to a decrease in hepatic alanine delivery. During recovery, hepatic blood flow was restored to basal, causing an increase in hepatic alanine delivery beyond exercise rates but still below resting rates. Hepatic fractional alanine extraction increased from 0.26 +/- 0.02 at rest to 0.64 +/- 0.03 during exercise and remained elevated during recovery. Net hepatic alanine uptake was 2.5 +/- 0.2 mumol.kg-1.min-1 at rest and remained unchanged during exercise but was increased during recovery. The conversion rate of [ 14 C]alanine to glucose had increased by 248 +/- 38% by 150 min of exercise and had increased further during recovery. The efficiency with which alanine was channeled into glucose in the liver was accelerated to a rate of 338 +/- 55% above basal by 150 min of exercise but declined slightly during recovery. In conclusion, 1) gluconeogenesis from alanine is accelerated during exercise, due to an increase in the hepatic fractional extraction of the amino acid and through intrahepatic mechanisms that more efficiently channel it into glucose

Mitochondrial defects associated with β-alanine toxicity: relevance to hyper-beta-alaninemia

Science.gov (United States)

Shetewy, Aza; Shimada-Takaura, Kayoko; Warner, Danielle; Jong, Chian Ju; Mehdi, Abu-Bakr Al; Alexeyev, Mikhail; Takahashi, Kyoko; Schaffer, Stephen W.

2016-01-01

Hyper-beta-alaninemia is a rare metabolic condition that results in elevated plasma and urinary β-alanine levels and is characterized by neurotoxicity, hypotonia, and respiratory distress. It has been proposed that at least some of the symptoms are caused by oxidative stress; however, only limited information is available on the mechanism of reactive oxygen species generation. The present study examines the hypothesis that β-alanine reduces cellular levels of taurine, which are required for normal respiratory chain function; cellular taurine depletion is known to reduce respiratory function and elevate mitochondrial superoxide generation. To test the taurine hypothesis, isolated neonatal rat cardiomyocytes and mouse embryonic fibroblasts were incubated with medium lacking or containing β-alanine. β-alanine treatment led to mitochondrial superoxide accumulation in conjunction with a decrease in oxygen consumption. The defect in β-alanine-mediated respiratory function was detected in permeabilized cells exposed to glutamate/malate but not in cells utilizing succinate, suggesting that β-alanine leads to impaired complex I activity. Taurine treatment limited mitochondrial superoxide generation, supporting a role for taurine in maintaining complex I activity. Also affected by taurine is mitochondrial morphology, as β-alanine-treated fibroblasts undergo fragmentation, a sign of unhealthy mitochondria that is reversed by taurine treatment. If left unaltered, β-alanine-treated fibroblasts also undergo mitochondrial apoptosis, as evidenced by activation of caspases 3 and 9 and the initiation of the mitochondrial permeability transition. Together, these data show that β-alanine mediates changes that reduce ATP generation and enhance oxidative stress, factors that contribute to heart failure. PMID:27023909

Crosslinked Aspartic Acids as Helix-Nucleating Templates.

Science.gov (United States)

Zhao, Hui; Liu, Qi-Song; Geng, Hao; Tian, Yuan; Cheng, Min; Jiang, Yan-Hong; Xie, Ming-Sheng; Niu, Xiao-Gang; Jiang, Fan; Zhang, Ya-Ou; Lao, Yuan-Zhi; Wu, Yun-Dong; Xu, Nai-Han; Li, Zi-Gang

2016-09-19

Described is a facile helix-nucleating template based on a tethered aspartic acid at the N-terminus [terminal aspartic acid (TD)]. The nucleating effect of the template is subtly influenced by the substituent at the end of the side-chain-end tether as indicated by circular dichroism, nuclear magnetic resonance, and molecular dynamics simulations. Unlike most nucleating strategies, the N-terminal amine is preserved, thus enabling further modification. Peptidomimetic estrogen receptor modulators (PERMs) constructed using this strategy show improved therapeutic properties. The current strategy can be regarded as a good complement to existing helix-stabilizing methods. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

The features of radiation damages in L-alanine crystals

International Nuclear Information System (INIS)

Zaitov, V.R.; Onischuk, V.A.

1996-01-01

The method of the ESR alanine dosimetry has appeared the most convenient one for measurement of radiation dose in the range 1-10 6 Gy. Its peculiarities are the wide dose range, the high accuracy, the absence fading at room temperature, the possibility of many times repeated measurements as dosemeter accumulates dose, the simplicity of measurements. Because of this performance ESR alanine dosimetry technique can be applied to continuous monitoring radiation doses absorbed by materials on nuclear power stations as well as of dose fields and restoration doses after an accident situation. In order to determine accurately the absorbed dose in an accident on background of accumulated dose for previous period, it is necessary to the utmost increase the accuracy of dosimetry system. For this reason it is necessary to know how the properties of free radicals which formings in irradiated L-alanine are displayed in signal ESR. With the purpose to detect the structure of the free radicals the ESR spectra the L-alanine and L-alanine-d 3 single crystals were studied. The samples were grown by slow evaporation of the saturated aqueous solution. For obtain the L-alanine-d 3 the three-divisible recrystallization in heavy water had been used. The samples were irradiated with 60 Co at room temperature and in liquid nitrogen. The irradiation doses were 10 kGy and dose rate was 8,3 Gy/s. To increase the resolution of the ESR spectra hyperfine structure the second derivative for the absorption curve was registered. The measurements were conducted in X-range at temperatures 77-430 K. (author)

Genipin-cross-linked poly(L-lysine)-based hydrogels: synthesis, characterization, and drug encapsulation.

Science.gov (United States)

Wang, Steven S S; Hsieh, Ping-Lun; Chen, Pei-Shan; Chen, Yu-Tien; Jan, Jeng-Shiung

2013-11-01

Genipin-cross-linked hydrogels composed of biodegradable and pH-sensitive cationic poly(L-lysine) (PLL), poly(L-lysine)-block-poly(L-alanine) (PLL-b-PLAla), and poly(L-lysine)-block-polyglycine (PLL-b-PGly) polypeptides were synthesized, characterized, and used as carriers for drug delivery. These polypeptide hydrogels can respond to pH-stimulus and their gelling and mechanical properties, degradation rate, and drug release behavior can be tuned by varying polypeptide composition and cross-linking degree. Comparing with natural polymers, the synthetic polypeptides with well-defined chain length and composition can warrant the preparation of the hydrogels with tunable properties to meet the criteria for specific biomedical applications. These hydrogels composed of natural building blocks exhibited good cell compatibility and enzyme degradability and can support cell attachment/proliferation. The evaluation of these hydrogels for in vitro drug release revealed that the controlled release profile was a biphasic pattern with a mild burst release and a moderate release rate thereafter, suggesting the drug molecules were encapsulated inside the gel matrix. With the versatility of polymer chemistry and conjugation of functional moieties, it is expected these hydrogels can be useful for biomedical applications such as polymer therapeutics and tissue engineering. Copyright © 2013 Elsevier B.V. All rights reserved.

Asymmetric adsorption of alanine by quartz powder from ethanol solution

Energy Technology Data Exchange (ETDEWEB)

Furuyama, Shozo; Sawada, Michio; Hachiya, Kinji; Morimoto, Tetsuo (Okayama Univ. (Japan). Faculty of Science)

1982-11-01

The asymmetric adsorption of the racemic alanine by the optically active quartz from ethanol solution at 8/sup 0/C was studied by the /sup 14/C-tracer method and the newly developed /sup 14/C-tracer ninhydrin-colorimetry combination method. The preferential adsorption of L-alanine by levorotatory quartz (l-quartz) and D-alanine by dextrorotatory quartz (d-quartz) was confirmed. The asymmetric adsorptivity (Asub(s)) falls in the range of 1.1 - 1.3, which is comparable with the value determined at - 80/sup 0/C in the previous paper. The effects of water content in the ethanol solution and of the adsorption temperature upon the adsorption affinity of alanine to quartz were also measured. The cause for the asymmetric adsorption is discussed from the crystallographic point of view.

On the helix equation

Directory of Open Access Journals (Sweden)

Taouil Hajer

2012-08-01

Full Text Available This paper is devoted to the helices processes, i.e. the solutions H : ℝ × Ω → ℝd, (t, ω ↦ H(t, ω of the helix equation egin{eqnarray} H(0,o=0; quad H(s+t,o= H(s,Phi(t,o +H(t,oonumber end{eqnarray} H ( 0 ,ω = 0 ;   H ( s + t,ω = H ( s, Φ ( t,ω + H ( t,ω where Φ : ℝ × Ω → Ω, (t, ω ↦ Φ(t, ω is a dynamical system on a measurable space (Ω, ℱ. More precisely, we investigate dominated solutions and non differentiable solutions of the helix equation. For the last case, the Wiener helix plays a fundamental role. Moreover, some relations with the cocycle equation defined by Φ, are investigated. Ce papier est consacré aux hélices, c’est-à-dire les solutions H : ℝ × Ω → ℝd, (t, ω ↦ H(t, ω de l’équation fonctionnelle egin{eqnarray} H(0,o=0; quad H(s+t,o= H(s,Phi(t,o +H(t,o onumber end{eqnarray} H ( 0 ,ω = 0 ;   H ( s + t,ω = H ( s, Φ ( t,ω + H ( t,ω où Φ : ℝ × Ω → Ω, (t, ω ↦ Φ(t, ω est un système dynamique défini sur un espace mesurable (Ω, ℱ. Plus présisément, nous déterminons d’abord les hélices dominées puis nous caractérisons les hélices non différentiables. Dans ce dernier cas, l’hélice de Wiener joue un rôle important. Nous précisons aussi quelques relations des hélices avec les cocycles définis par Φ.

Genome-wide identification and analysis of the chicken basic helix-loop-helix factors.

Science.gov (United States)

Liu, Wu-Yi; Zhao, Chun-Jiang

2010-01-01

Members of the basic helix-loop-helix (bHLH) family of transcription factors play important roles in a wide range of developmental processes. In this study, we conducted a genome-wide survey using the chicken (Gallus gallus) genomic database, and identified 104 bHLH sequences belonging to 42 gene families in an effort to characterize the chicken bHLH transcription factor family. Phylogenetic analyses revealed that chicken has 50, 21, 15, 4, 8, and 3 bHLH members in groups A, B, C, D, E, and F, respectively, while three members belonging to none of these groups were classified as ''orphans". A comparison between chicken and human bHLH repertoires suggested that both organisms have a number of lineage-specific bHLH members in the proteomes. Chromosome distribution patterns and phylogenetic analyses strongly suggest that the bHLH members should have arisen through gene duplication at an early date. Gene Ontology (GO) enrichment statistics showed 51 top GO annotations of biological processes counted in the frequency. The present study deepens our understanding of the chicken bHLH transcription factor family and provides much useful information for further studies using chicken as a model system.

A genome-wide survey on basic helix-loop-helix transcription factors in giant panda.

Directory of Open Access Journals (Sweden)

Chunwang Dang

Full Text Available The giant panda (Ailuropoda melanoleuca is a critically endangered mammalian species. Studies on functions of regulatory proteins involved in developmental processes would facilitate understanding of specific behavior in giant panda. The basic helix-loop-helix (bHLH proteins play essential roles in a wide range of developmental processes in higher organisms. bHLH family members have been identified in over 20 organisms, including fruit fly, zebrafish, mouse and human. Our present study identified 107 bHLH family members being encoded in giant panda genome. Phylogenetic analyses revealed that they belong to 44 bHLH families with 46, 25, 15, 4, 11 and 3 members in group A, B, C, D, E and F, respectively, while the remaining 3 members were assigned into "orphan". Compared to mouse, the giant panda does not encode seven bHLH proteins namely Beta3a, Mesp2, Sclerax, S-Myc, Hes5 (or Hes6, EBF4 and Orphan 1. These results provide useful background information for future studies on structure and function of bHLH proteins in the regulation of giant panda development.

Determination of the torsion angles of alanine and glycine residues of model compounds of spider silk (AGG){sub 10} using solid-state NMR methods

Energy Technology Data Exchange (ETDEWEB)

Ashida, Jun; Ohgo, Kosuke; Komatsu, Kohei; Kubota, Ayumi; Asakura, Tetsuo [Tokyo University of Agriculture and Technology, Department of Biotechnology (Japan)], E-mail: asakura@cc.tuat.ac.jp

2003-02-15

Spiders synthesize several kinds of silk fibers. In the primary structure of spider silk, one of the major ampullate (dragline, frame) silks, spidroin 1, and flagelliform silk (core fibers of adhesive spiral), there are common repeated X-Gly-Gly (X = Ala, Leu, Pro, Tyr, Glu, and Arg) sequences, which are considered to be related to the elastic character of these fibers. In this paper, two dimensional spin diffusion solid-state NMR under off magic angle spinning (OMAS), {sup 13}C chemical shift contour plots, and Rotational Echo DOuble Resonance (REDOR) were applied to determine the torsion angles of one Ala and two kinds of Gly residues in the Ala-Gly-Gly sequence of {sup 13}C=O isotope-labeled (Ala-Gly-Gly){sub 10}. The torsion angles were determined to be ({phi}, {psi}) = (-90 deg., 150 deg.) within an experimental error of {+-}10 deg. for each residue. This conformation is characterized as 3{sub 1} helix which is in agreement with the structure proposed from the X-ray powder diffraction pattern of poly(Ala-Gly-Gly). The 3{sub 1} helix of (Ala-Gly-Gly){sub 10} does not change by formic acid treatment although (Ala-Gly){sub 15} easily changes from the silk I conformation (the structure of Bombyx mori silk fibroin before spinning in the solid state) to silk II conformation (the structure of the silk fiber after spinning) by such treatment. Thus, the 3{sub 1} helix conformation of (Ala-Gly-Gly){sub 10} is considered very stable. Furthermore, the torsion angles of the 16th Leu residue of (Leu-Gly-Gly){sub 10} were also determined as ({phi}, {psi}) = (-90 deg., 150 deg.) and this peptide is also considered to take 3{sub 1} helix conformation.

Organizing product innovation: hierarchy, market or triple-helix networks?

Science.gov (United States)

Fitjar, Rune Dahl; Gjelsvik, Martin; Rodríguez-Pose, Andrés

This paper assesses the extent to which the organization of the innovation effort in firms, as well as the geographical scale at which this effort is pursued, affects the capacity to benefit from product innovations. Three alternative modes of organization are studied: hierarchy, market and triple-helix-type networks. Furthermore, we consider triple-helix networks at three geographical scales: local, national and international. These relationships are tested on a random sample of 763 firms located in five urban regions of Norway which reported having introduced new products or services during the preceding 3 years. The analysis shows that firms exploiting internal hierarchy or triple-helix networks with a wide range of partners managed to derive a significantly higher share of their income from new products, compared to those that mainly relied on outsourcing within the market. In addition, the analysis shows that the geographical scale of cooperation in networks, as well as the type of partner used, matters for the capacity of firms to benefit from product innovation. In particular, firms that collaborate in international triple-helix-type networks involving suppliers, customers and R&D institutions extract a higher share of their income from product innovations, regardless of whether they organize the processes internally or through the network.

Poly-P storage by natural biofilms in streams with varying biogeochemistry

Science.gov (United States)

Carrick, H. J.

2015-12-01

Anthropogenic inputs of nitrogen (N) and phosphorus (P) have increased in many watersheds throughout the world; these inputs have been linked to the eutrophication of inland and coastal waters worldwide. We selected and surveyed 20, third-order streams that supported a range of water column biogeochemical conditions (conductivity, nutrient concentrations) located in the mid-Atlantic region, USA. Biofilm biomass, algal taxonomic composition, and nutrient stoichiometry (C, N, P, and poly-P) were measured at all stream sites. Pulse-amplitude modulation fluorometry (PAM) was used to estimate photosynthetic parameters for stream biofilms (e.g., alpha, Pmax), while microbiology techniques were used to verify poly-P storage by pro- and eukaryotic components of the biofilm (e.g., epi-fluorescent staining). As anticipated, chlorophyll ranged over 2 orders of magnitude among the streams (range 10-1,000 mg/m2). Biofilm chlorophyll and algal biovolume levels increased with water column nutrient contents, while the C:P ratio within the biofilm decreased. Both pro and eukaryotic organisms were present in resident biofilms and actively stored intracellular poly-P. Finally, the rate of photosynthetic within the biofilms appeared to be driven the nutritional condition of the biofilms; pmax and alpha values increased with significantly with stream biofilm poly-P content (r2 = 0.35 and 0.44, respectively). These results indicated that where nutrients are plentiful, biofilms P storage is favored, and this is likely a key regulator of stream biofilm biomass and productivity.

A new synthesis of [3-11C]pyruvic acid using alanine racemase

International Nuclear Information System (INIS)

Ikemoto, M.; Okamoto, E.; Sasaki, M.; Haradahira, T.; Omura, H.; Furuya, Y.; Suzuki, K.; Watanabe, Y.

1998-01-01

The synthesis of [3- 11 C]pyruvic acid was attempted by two reaction systems (A: alanine racemase and D-amino acid oxidase, B: alanine racemase and L-alanine dehydrogenase) utilizing a new thermostable enzyme, alanine racemase. Conversion rates from D,L-[3- 11 C]alanine to [3- 11 C]pyruvic acid were almost 100% in both methods. Similar results were obtained with immobilized enzymes packed in a single column. Furthermore, the same column could be used repeatedly without a remarkable decrease of the [3- 11 C]pyruvic acid yield. Various matrices were tested for the immobilizing enzyme, and Aminopropyl-CPG was concluded to be the most suitable since the loss of the enzyme activity was the least in the studied matrices

Uses of polymer-alanine film/ESR dosimeters in dosimetry of ionizing radiation

International Nuclear Information System (INIS)

Xie Liqing; Zhang Yinfeng; Dai Jinxian; Lu Ting; Chen Ruyi; Yang Hua

1993-01-01

Alanine ESR dosimetry is a reliable method, used in a various fields of ionizing radiation. The polymer-alanine film/ESR dosimeters of 0.3 -0.4 mm thickness were prepared and their dosimetric properties were studied for 60 Co γ photons and 3 - 5 MeV electrons in the dose range from 20 Gy to 100 kGy. The results show that under normal conditions the alanine calibration curves are linear in the dose range from 100 Gy to 10kGy. The dose profiles at the electron radiation field were measured with the film alanine dosimeters. The polymer-alanine film dosimeters were used for ion implantation of 400 keV ion implantor. Their dose response and energy dependence were investigated initially. (Author)

HIF-1{alpha} is necessary to support gluconeogenesis during liver regeneration

Energy Technology Data Exchange (ETDEWEB)

Tajima, Toshihide [Department of Obstetrics and Gynecology, School of Medicine, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582 (Japan); Goda, Nobuhito, E-mail: goda@waseda.jp [Department of Life Science and Medical Bio-Science, School of Advanced Science and Engineering, Waseda University, TWIns 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo 162-8480 (Japan); Fujiki, Natsuko; Hishiki, Takako; Nishiyama, Yasumasa [Department of Biochemistry and Integrative Medical Biology, School of Medicine, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582 (Japan); Senoo-Matsuda, Nanami [Department of Life Science and Medical Bio-Science, School of Advanced Science and Engineering, Waseda University, TWIns 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo 162-8480 (Japan); Shimazu, Motohide [Department of Surgery, Tokyo Medical University Hachioji Medical Center, 1163 Tatemachi, Hachioji, Tokyo 193-0998 (Japan); Soga, Tomoyoshi [The Institute for Advanced Biosciences, Keio University, Tsuruoka City, Yamagata 997-0052 (Japan); Yoshimura, Yasunori [Department of Obstetrics and Gynecology, School of Medicine, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582 (Japan); Johnson, Randall S. [Molecular Biology Section, Division of Biology, University of California, San Diego, La Jolla, CA 92093 (United States); Suematsu, Makoto [Department of Biochemistry and Integrative Medical Biology, School of Medicine, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582 (Japan)

2009-10-02

Coordinated recovery of hepatic glucose metabolism is prerequisite for normal liver regeneration. To examine roles of hypoxia inducible factor-1{alpha} (HIF-1{alpha}) for hepatic glucose homeostasis during the reparative process, we inactivated the gene in hepatocytes in vivo. Following partial hepatectomy (PH), recovery of residual liver weight was initially retarded in the mutant mice by down-regulation of hepatocyte proliferation, but occurred comparably between the mutant and control mice at 72 h after PH. At this time point, the mutant mice showed lowered blood glucose levels with enhanced accumulation of glycogen in the liver. The mutant mice exhibited impairment of hepatic gluconeogenesis as assessed by alanine tolerance test. This appeared to result from reduced expression of PGK-1 and PEPCK since 3-PG, PEP and malate were accumulated to greater extents in the regenerated liver. In conclusion, these findings provide evidence for roles of HIF-1{alpha} in the regulation of gluconeogenesis under liver regeneration.

Thin layer alanine dosimeter with optical spectrophotometric evaluation

International Nuclear Information System (INIS)

Zagorski, Z.P.

2000-01-01

Experience in the high dose dosimetry of gamma radiation, gathered in our group from the sixties till now, allows to express the opinion, that techniques applied are adequate to solve problems. It can be confirmed by the fact that 60% of laboratories participating in the international comparison during the duration of the contract obtained satisfactory results. Adaptation of these methods, in particular of the alanine-ESR dosimetry to highly inhomogeneous fields of EB gives poor results, as it has been shown on thin films of the alanine/polymer composite. However, the applications of these films give excellent results if the concentration of the radical CH 3 C·H CO 2 - is measured by diffuse reflection spectrophotometry, which tolerates poor transparency of the composite and is insensitive to the orientation of crystals of alanine in thin films, what is disqualifying the ESR measurements. The development of thin-film dosimeters for EB processing was possible due to new developments in solid state radiation chemistry. The research has revealed some unsolved questions, e.g. of the high temperature coefficient of alanine based dosimeters, of the role of the size of spurs and the necessity to adapt dosimetry to the energy spectrum of electrons, because every type of accelerators differs in that respect. (author)

Alanine turnover in the postabsorptive state and during parenteral hyperalimentation before and after surgery

NARCIS (Netherlands)

Sauerwein, H. P.; Michels, R. P.; Cejka, V.

1981-01-01

Influence of total parenteral nutrition and operation on alanine turnover and venous alanine concentration was determined in 5 patients with stomach carcinoma using single technique of U-14C alanine. Every patient served at his own control. In the postabsorptive state alanine turnover was 1.63 +/-

Murine elongation factor 1 alpha (EF-1 alpha) is posttranslationally modified by novel amide-linked ethanolamine-phosphoglycerol moieties. Addition of ethanolamine-phosphoglycerol to specific glutamic acid residues on EF-1 alpha

International Nuclear Information System (INIS)

Whiteheart, S.W.; Shenbagamurthi, P.; Chen, L.; Cotter, R.J.; Hart, G.W.

1989-01-01

Elongation Factor 1 alpha (EF-1 alpha), an important eukaryotic translation factor, transports charged aminoacyl-tRNA from the cytosol to the ribosomes during poly-peptide synthesis. Metabolic radiolabeling with [ 3 H] ethanolamine shows that, in all cells examined, EF-1 alpha is the major radiolabeled protein. Radiolabeled EF-1 alpha has an apparent Mr = 53,000 and a basic isoelectric point. It is cytosolic and does not contain N-linked oligosaccharides. Trypsin digestion of murine EF-1 alpha generated two major [ 3 H]ethanolamine-labeled peptides. Three peptides were sequenced and were identical to two distinct regions of the human EF-1 alpha protein. Blank sequencing cycles coinciding with glutamic acid in the human cDNA-derived sequence were also found to release [ 3 H]ethanolamine, and compositional analysis of these peptides confirmed the presence of glutamic acid. Dansylation analysis demonstrates that the amine group of the ethanolamine is blocked. These results indicate that EF-1 alpha is posttranslationally modified by the covalent attachment of ethanolamine via an amide bond to at least two specific glutamic acid residues (Glu-301 and Glu-374). The hydroxyl group of the attached ethanolamine was shown by mass spectrometry and compositional analysis, to be further modified by the addition of a phosphoglycerol unit. This novel posttranslational modification may represent an important alteration of EF-1 alpha, comparable to the regulatory effects of posttranslational methylation of EF-1 alpha lysine residues

Role of L-alanine for redox self-sufficient amination of alcohols.

Science.gov (United States)

Klatte, Stephanie; Wendisch, Volker F

2015-01-23

In white biotechnology biocatalysis represents a key technology for chemical functionalization of non-natural compounds. The plasmid-born overproduction of an alcohol dehydrogenase, an L-alanine-dependent transaminase and an alanine dehydrogenase allows for redox self-sufficient amination of alcohols in whole cell biotransformation. Here, conditions to optimize the whole cell biocatalyst presented in (Bioorg Med Chem 22:5578-5585, 2014), and the role of L-alanine for efficient amine functionalization of 1,10-decanediol to 1,10-diaminodecane were analyzed. The enzymes of the cascade for amine functionalization of alcohols were characterized in vitro to find optimal conditions for an efficient process. Transaminase from Chromobacterium violaceum, TaCv, showed three-fold higher catalytic efficiency than transaminase from Vibrio fluvialis, TaVf, and improved production at 37°C. At 42°C, TaCv was more active, which matched thermostable alcohol dehydrogenase and alanine dehydrogenase and improved the 1,10-diaminodecane production rate four-fold. To study the role of L-alanine in the whole cell biotransformation, the L-alanine concentration was varied and 1,10.diaminodecane formation tested with constant 10 mM 1,10- decanediol and 100 mM NH4Cl. Only 5.6% diamine product were observed without added L-alanine. L-alanine concentrations equimolar to that of the alcohol enabled for 94% product formation but higher L-alanine concentrations allowed for 100% product formation. L-alanine was consumed by the E. coli biocatalyst, presumably due to pyruvate catabolism since up to 16 mM acetate accumulated. Biotransformation employing E. coli strain YYC202/pTrc99a-ald-adh-ta Cv, which is unable to catabolize pyruvate, resulted in conversion with a selectivity of 42 mol-%. Biotransformation with E. coli strains only lacking pyruvate oxidase PoxB showed similar reduced amination of 1,10-decanediol indicating that oxidative decarboxylation of pyruvate to acetate by PoxB is primarily

Preparation of alanine/ESR dosimeter using different binder of polymer blend

International Nuclear Information System (INIS)

Razzak, M.T.; Sudiro, Sutjipto; Sudradjat, Adjat; Waskito, Ashar; Djamili, M.F.

1995-01-01

Different composition of polymer blend of low density polyethylene (PE) and polystyrene (PS) have been studied to be used as a binder for the preparation of Alanine/ESR dosimeter. The polymer binder and Alanine powder were blended in Laboplastomil Mixer at 140 o C and then it was pressed into a plastic film of 0.50 mm thickness. The film was cut into sample size of 250 mm x 2.5 mm and irradiated by gamma rays from a cobalt-60 source at different dose and dose rate. It was found that a blend of Alanine, PS and PE in composition of 60:30:10 is appropriate to prepare the Alanine/ESR dosimeter. (author)

Dynamics of solid alanine by means of nuclear magnetic resonance relaxometry

Science.gov (United States)

Kubica-Misztal, A.; Rochowski, P.; Florek-Wojciechowska, M.; Kruk, D.

2017-04-01

1H nuclear magnetic resonance relaxometry was applied to investigate the dynamics of l-alanine in the solid phase (powder). The experimental studies were carried out in a very broad frequency range, covering four orders of magnitude—from 4 kHz to 40 MHz (referring to the 1H resonance frequency) in order to probe motional processes of much different time scales by a single experiment. To get access to the dynamics of different proton groups of alanine, the 1H spin-lattice relaxation measurements were performed for non-deuterated and partially deuterated alanine. The experiments were carried out in the temperature range of 293 K-370 K (non-deuterated alanine) and 318 K-370 K (partially deuterated alanine). As a result of a thorough theoretical analysis of the extensive set of experimental results, three motional processes occurring on different time scales are identified and quantitatively described. The slowest process occurs on a time scale of μs and it is attributed to the collective dynamics of a 3D hydrogen bond network of alanine, while the intermediate, attributed to the dynamics of the NH3 group, corresponds to the range of tenths of ns. The fast process describes the rotation of the CH3 group.

Characterization of the l-alanine exporter AlaE of Escherichia coli and its potential role in protecting cells from a toxic-level accumulation of l-alanine and its derivatives

OpenAIRE

Kim, Seryoung; Ihara, Kohei; Katsube, Satoshi; Hori, Hatsuhiro; Ando, Tasuke; Isogai, Emiko; Yoneyama, Hiroshi

2015-01-01

We previously reported that the alaE gene of Escherichia coli encodes the l-alanine exporter AlaE. The objective of this study was to elucidate the mechanism of the AlaE exporter. The minimum inhibitory concentration of l-alanine and l-alanyl-l-alanine in alaE-deficient l-alanine-nonmetabolizing cells MLA301ΔalaE was 4- and >4000-fold lower, respectively, than in the alaE-positive parent cells MLA301, suggesting that AlaE functions as an efflux pump to avoid a toxic-level accumulation of intr...

Determination of ammonium ion using a reagentless amperometric biosensor based on immobilized alanine dehydrogenase.

Science.gov (United States)

Tan, Ling Ling; Musa, Ahmad; Lee, Yook Heng

2011-01-01

The use of the enzyme alanine dehydrogenase (AlaDH) for the determination of ammonium ion (NH(4)(+)) usually requires the addition of pyruvate substrate and reduced nicotinamide adenine dinucleotide (NADH) simultaneously to effect the reaction. This addition of reagents is inconvenient when an enzyme biosensor based on AlaDH is used. To resolve the problem, a novel reagentless amperometric biosensor using a stacked methacrylic membrane system coated onto a screen-printed carbon paste electrode (SPE) for NH(4)(+) ion determination is described. A mixture of pyruvate and NADH was immobilized in low molecular weight poly(2-hydroxyethyl methacrylate) (pHEMA) membrane, which was then deposited over a photocured pHEMA membrane (photoHEMA) containing alanine dehydrogenase (AlaDH) enzyme. Due to the enzymatic reaction of AlaDH and the pyruvate substrate, NH(4)(+) was consumed in the process and thus the signal from the electrocatalytic oxidation of NADH at an applied potential of +0.55 V was proportional to the NH(4)(+) ion concentration under optimal conditions. The stacked methacrylate membranes responded rapidly and linearly to changes in NH(4)(+) ion concentrations between 10-100 mM, with a detection limit of 0.18 mM NH(4)(+) ion. The reproducibility of the amperometrical NH(4)(+) biosensor yielded low relative standard deviations between 1.4-4.9%. The stacked membrane biosensor has been successfully applied to the determination of NH(4)(+) ion in spiked river water samples without pretreatment. A good correlation was found between the analytical results for NH(4)(+) obtained from the biosensor and the Nessler spectrophotometric method.

Sensitivity comparison of two L-alanine doped blends to different photon energies

International Nuclear Information System (INIS)

Chen, Felipe; Vega Ramirez, Jose; Nicolucci, Patricia; Baffa, Oswaldo

2008-01-01

Full text: Blends of L-alanine (85% weight proportion) with KI (10%) and with PbI 2 (10%), these last two compounds acting as dopants, and with PVA (5%) acting as binder, were prepared in water at 80 C degrees. A blend of pure L-alanine (95%) with PVA (5%) was also prepared. The three blends were irradiated with photon beams of different energies (120 kV, 60 Co and 10 MV) with a unique dose of 30 Gy to compare their sensitivities for those three energies. EPR spectra of the three irradiated blends were recorded in a K-Band spectrometer (24 GHz) taking aliquots of about 4 mg for each blend. The energy sensitivity of a blend was defined as the peak-to-peak amplitude of its EPR spectrum central line. For the 60 Co energy (1.25 MeV) the blends presented practically the same sensitivity indicating that the presence of the dopants does not affect the sensitivity of L-alanine. For 10 MV X-rays there was an increment (around 20% - 30 %) in sensitivity for the two L-alanine doped blends compared with the pure L-alanine blend (not doped). In the case of 120 kV X-rays, the blends ala+KI and ala+PbI 2 showed an increment of 10 and 20 times, respectively, more sensitivity than the pure L-alanine blend. It is concluded that the dopants KI and PbI 2 produce a great enhance of the L-alanine sensitivity to low-energy photons. For the same dopant's content (10%) in the blend, PbI 2 showed a better performance. These results encourage us to try to enhance the sensitivity of L-alanine even more increasing the dopant's content in the blend. Application of these L-alanine doped blends in the dosimetry in diagnostic radiology could be possible. (author)

Postirradiation effects in alanine dosimeter probes of two different suppliers

International Nuclear Information System (INIS)

Anton, Mathias

2008-01-01

The measurand relevant for the dosimetry for radiation therapy is the absorbed dose to water, D W . The Physikalisch-Technische Bundesanstalt (PTB) is establishing a secondary standard for D W for high-energy photon and electron radiation based on electron spin resonance (ESR) of the amino acid alanine. For practical applications, like, for example, intercomparison measurements using the ESR/alanine dosimetry system, the temporal evolution of the ESR signal of irradiated probes is an important issue. This postirradiation behaviour is investigated for alanine pellets of two different suppliers for different storage conditions. The influence of the storage conditions on the temporal evolution may be dependent on the type of probes used. The measurement and analysis method developed at the PTB is able to circumvent the apparent difficulties in the case of alanine/paraffin probes. Care has to be taken in case this method cannot be applied

Double helix vortex breakdown in a turbulent swirling annular jet flow

NARCIS (Netherlands)

Vanierschot, M.; Perçin, M.; van Oudheusden, B.W.

2018-01-01

In this paper, we report on the structure and dynamics of double helix vortex breakdown in a turbulent annular swirling jet. Double helix breakdown has been reported previously for the laminar flow regime, but this structure has rarely been observed in turbulent flow. The flow field is

EPR structure of the gamma irradiated alanine spectrum

Energy Technology Data Exchange (ETDEWEB)

Cabral P, A; Jimenez D, H; Urena N, F; Galindo, S; Bosch, P

1992-03-15

In this study is shown that the broadened five-line EPR pattern of the gamma irradiated alanine possibly decomposes into a more complex pattern when the recorded spectrum is subject to an operation of deconvolution. The EPR powder spectra of gamma irradiated DL- and L-alanine with and without binders are analysed. In all recorded spectra, each observed line is resolved into an asymmetrical triplet when a Gaussian distribution of 8.2 gauss width is removed, by deconvolution, from the observed spectrum. On the other hand, from a simple fitting analysis carried out on the original data, one encounters that some calculated relations between characteristic parameters, such as intensity ratios, deviate consistently from assumed height ratios. Both, from deconvolution and fitting results, a different structure is suggested for the observed broadened five-line EPR pattern of {gamma}-irradiated powder DL- and L-alanine. (Author)

EPR structure of the gamma irradiated alanine spectrum

International Nuclear Information System (INIS)

Cabral P, A.; Jimenez D, H.; Urena N, F.; Galindo, S.; Bosch, P.

1992-03-01

In this study is shown that the broadened five-line EPR pattern of the gamma irradiated alanine possibly decomposes into a more complex pattern when the recorded spectrum is subject to an operation of deconvolution. The EPR powder spectra of gamma irradiated DL- and L-alanine with and without binders are analysed. In all recorded spectra, each observed line is resolved into an asymmetrical triplet when a Gaussian distribution of 8.2 gauss width is removed, by deconvolution, from the observed spectrum. On the other hand, from a simple fitting analysis carried out on the original data, one encounters that some calculated relations between characteristic parameters, such as intensity ratios, deviate consistently from assumed height ratios. Both, from deconvolution and fitting results, a different structure is suggested for the observed broadened five-line EPR pattern of γ-irradiated powder DL- and L-alanine. (Author)

Introduction of a proline residue into position 31 of the loop of the dimeric 4-alpha-helical protein ROP causes a drastic destabilization.

Science.gov (United States)

Peters, K; Hinz, H J; Cesareni, G

1997-10-01

The exchange of an alanine with a proline residue in position 31 of the loop region of the dimeric 4-alpha-helical-bundle protein ROP causes a reduction in the alpha-helix content of 7% and a reduction in stability of about 40% compared to the wild type parameters. The Gibbs energy of unfolding by denaturants extrapolated linearly to zero denaturant concentration, delta G0D (buffer, 25 degrees C), has been determined to be 43 kJ (mol dimer)-1. The corresponding ROPwt value is 72 kJ (mol dimer)-1 (Steif et al., 1993). The extrapolated delta G0D values obtained from urea and GdmHCI un- and refolding studies are identical within error limits. Deconvolution of the stability values into enthalpy and entropy terms resulted in the following parameters. At T1/2 = 43 degrees C (Cprotein = 0.05 mg.ml-1) the ROP A31P mutant is characterized by delta Hv.H.0 = 272 kJ (mol dimer)-1, delta Cp = 7.2 kJ (mol dimer)-1 K-1, delta S0 = 762 J (mol dimer)-1 K-1. These parameters are only approximately 50% as large as the corresponding values of ROPwt. We assume that the significant reduction in stability reflects the absence of at least one hydrogen bond as well as deformation of the protein structure. This interpretation is supported by the reduction in the change in heat capacity observed for the A31P mutant relative to ROPwt, by the increased aggregation tendency of the mutant and by the reduced specific CD absorption at 222 nm. All results support the view that in the case of ROP protein the loop region plays a significant role in the maintenance of native structure and conformational stability.

Brain alanine formation as an ammonia-scavenging pathway during hyperammonemia

DEFF Research Database (Denmark)

Dadsetan, Sherry; Kukolj, Eva; Bak, Lasse Kristoffer

2013-01-01

(MSO) enhances incorporation of (15)NH4(+) in alanine during acute hyperammonemia. We observed a fourfold increased amount of (15)NH4 incorporation in brain alanine in rats treated with MSO. Furthermore, co-cultures of neurons and astrocytes exposed to (15)NH4Cl in the absence or presence of MSO...... demonstrated a dose-dependent incorporation of (15)NH4 into alanine together with increased (15)N incorporation in glutamate. These findings provide evidence that ammonia is detoxified by the concerted action of GDH and ALAT both in vivo and in vitro, a mechanism that is accelerated in the presence of MSO...

Isotope effect of optical activity measurements on L-α-alanine

International Nuclear Information System (INIS)

Darge, W.; Laczko, I.; Thiemann, W.

1976-01-01

If an optically active organic substance is labelled in the chirality center with another isotopic species (such as 15 N for 14 N) a pronounced variation of rotatory power is predicted. It was tried to varify this idea experimentally on L-α-alanine and found an isotope effect in ORD (optical rotatory dispersion). The magnitude of the rotation is mainly dependent on the pH of the solvent. The ratio of the optical activity alanine- 14 N/alanine- 15 N is about 1.02. It can be seen that the ratios of the molecular rotations are consistently lower than the corresponding ratios of the specific rotations. This is of course due to the fact that the molecular mass 15 M is larger than 14 M. This means tthat the mass difference is already taken into account so that the ratio of the molecular rotations could be defined as the ''net'' isotope effect in the ORDs of 15 N-substitued alanine. From the fact the ORD is different for the isotope-substitued alanine, one can reasonably assume that the absorption coefficient is also different. This leads to speculations about certain problems in the chemical evolution of the biosphere, such as the origin of optical activity. (T.G.)

Unraveling the Role of the C-terminal Helix Turn Helix of the Coat-binding Domain of Bacteriophage P22 Scaffolding Protein*

Science.gov (United States)

Padilla-Meier, G. Pauline; Gilcrease, Eddie B.; Weigele, Peter R.; Cortines, Juliana R.; Siegel, Molly; Leavitt, Justin C.; Teschke, Carolyn M.; Casjens, Sherwood R.

2012-01-01

Many viruses encode scaffolding and coat proteins that co-assemble to form procapsids, which are transient precursor structures leading to progeny virions. In bacteriophage P22, the association of scaffolding and coat proteins is mediated mainly by ionic interactions. The coat protein-binding domain of scaffolding protein is a helix turn helix structure near the C terminus with a high number of charged surface residues. Residues Arg-293 and Lys-296 are particularly important for coat protein binding. The two helices contact each other through hydrophobic side chains. In this study, substitution of the residues of the interface between the helices, and the residues in the β-turn, by aspartic acid was used examine the importance of the conformation of the domain in coat binding. These replacements strongly affected the ability of the scaffolding protein to interact with coat protein. The severity of the defect in the association of scaffolding protein to coat protein was dependent on location, with substitutions at residues in the turn and helix 2 causing the most significant effects. Substituting aspartic acid for hydrophobic interface residues dramatically perturbs the stability of the structure, but similar substitutions in the turn had much less effect on the integrity of this domain, as determined by circular dichroism. We propose that the binding of scaffolding protein to coat protein is dependent on angle of the β-turn and the orientation of the charged surface on helix 2. Surprisingly, formation of the highly complex procapsid structure depends on a relatively simple interaction. PMID:22879595

Structure and Intramolecular Proton Transfer of Alanine Radical Cations

International Nuclear Information System (INIS)

Lee, Gab Yong

2012-01-01

The structures of the four lowest alanine conformers, along with their radical cations and the effect of ionization on the intramolecular proton transfer process, are studied using the density functional theory and MP2 method. The energy order of the radical cations of alanine differs from that of the corresponding neutral conformers due to changes in the basicity of the NH 2 group upon ionization. Ionization favors the intramolecular proton transfer process, leading to a proton-transferred radical-cation structure, [NH 3 + -CHCH 3 -COO·], which contrasts with the fact that a proton-transferred zwitterionic conformer is not stable for a neutral alanine in the gas phase. The energy barrier during the proton transfer process is calculated to be about 6 kcal/mol

Genome-Wide Identification and Analysis of the Chicken Basic Helix-Loop-Helix Factors

Directory of Open Access Journals (Sweden)

Wu-yi Liu

2010-01-01

Full Text Available Members of the basic helix-loop-helix (bHLH family of transcription factors play important roles in a wide range of developmental processes. In this study, we conducted a genome-wide survey using the chicken (Gallus gallus genomic database, and identified 104 bHLH sequences belonging to 42 gene families in an effort to characterize the chicken bHLH transcription factor family. Phylogenetic analyses revealed that chicken has 50, 21, 15, 4, 8, and 3 bHLH members in groups A, B, C, D, E, and F, respectively, while three members belonging to none of these groups were classified as ‘‘orphans’’. A comparison between chicken and human bHLH repertoires suggested that both organisms have a number of lineage-specific bHLH members in the proteomes. Chromosome distribution patterns and phylogenetic analyses strongly suggest that the bHLH members should have arisen through gene duplication at an early date. Gene Ontology (GO enrichment statistics showed 51 top GO annotations of biological processes counted in the frequency. The present study deepens our understanding of the chicken bHLH transcription factor family and provides much useful information for further studies using chicken as a model system.

Development of portable ESR spectrometer as a reader for alanine dosimeters

International Nuclear Information System (INIS)

Kojima, T.; Haruyama, Y.; Tachibana, H.; Tanaka, R.; Okamoto, J.

1993-01-01

A prototype portable electron spin resonance (ESR) spectrometer was designed and tested, and its feasibility as a reader of alanine dosimeters was studied from the two standpoints of reproducibility of readings and sensitivity sufficient for dosimetry in the absorbed dose range 1-100 kGy. It has two main components: a permanent magnet and resonator; and a unit box with a microwave and auto-frequency control (AFC) circuit, a sweep controller of magnetic field, display, etc. In the present preliminary study, reproducibility values are measured with the same ESR parameters and alanine-polystyrene (alanine-PS) dosimeter at a dose of 1 kGy: repeatedly measuring without removing dosimeter from the cavity; individual measurement with removing and inserting again into the cavity with readjustment of ESR parameters. Alanine/ESR dosimetry using this spectrometer has a measurable dose range from 1 to 100 kGy with relatively high precision within ± 3% (1σ) as a preliminary result. The portable ESR spectrometer may also be modified as an automatic, more precise, dedicated alanine dosimeter reader. (author)

d-Alanine metabolism is essential for growth and biofilm formation of Streptococcus mutans.

Science.gov (United States)

Qiu, W; Zheng, X; Wei, Y; Zhou, X; Zhang, K; Wang, S; Cheng, L; Li, Y; Ren, B; Xu, X; Li, Y; Li, M

2016-10-01

Part of the d-alanine (d-Ala) metabolic pathway in bacteria involves the conversion of l-alanine to d-Ala by alanine racemase and the formation of d-alanyl-d-alanine by d-alanine-d-alanine ligase, the product of which is involved in cell wall peptidoglycan synthesis. At present, drugs that target the metabolic pathway of d-Ala are already in clinical use - e.g. d-cycloserine (DCS) is used as an antibiotic against Mycobacterium tuberculosis. Streptococcus mutans is the main cariogenic bacterium in the oral cavity. Its d-Ala metabolism-associated enzymes alanine racemase and d-alanine-d-alanine ligase are encoded by the genes smu.1834 and smu.599, respectively, which may be potential targets for inhibitors. In this study, the addition of DCS blocked the d-Ala metabolic pathway in S. mutans, leading to bacterial cell wall defects, significant inhibition of bacterial growth and biofilm formation, and reductions in extracellular polysaccharide production and bacterial adhesion. However, the exogenous addition of d-Ala could reverse the inhibitory effect of DCS. Through the means of drug regulation, our study demonstrated, for the first time, the importance of d-Ala metabolism in the survival and biofilm formation of S. mutans. If the growth of S. mutans can be specifically inhibited by designing drugs that target d-Ala metabolism, then this may serve as a potential new treatment for dental caries. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Synthesis of silver nanoparticles using DL-alanine for ESR dosimetry applications

International Nuclear Information System (INIS)

Guidelli, Eder José; Ramos, Ana Paula; Zaniquelli, Maria Elisabete D.; Nicolucci, Patricia; Baffa, Oswaldo

2012-01-01

The potential use of alanine for the production of nanoparticles is presented here for the first time. Silver nanoparticles were synthesized using a simple green method, namely the thermal treatment of silver nitrate aqueous solutions with DL-alanine. The latter compound was employed both as a reducing and a capping agent. Particles with average size equal to 7.5 nm, face-centered cubic crystalline structure, narrow size distribution, and spherical shape were obtained. Interaction between the silver ions present on the surface of the nanoparticles and the amine group of the DL-alanine molecule seems to be responsible for reduction of the silver ions and for the stability of the colloid. The bio-hybrid nano-composite was used as an ESR dosimeter. The amount of silver nanoparticles in the nanocomposite was not sufficient to cause considerable loss of tissue equivalency. Moreover, the samples containing nanoparticles presented increased sensitivity and reduced energetic dependence as compared with pure DL-alanine, contributing to the construction of small-sized dosimeters. - Highlights: ► The synthesis is environmentally benign, easy to perform, and of low-cost. ► DL-Alanine was employed both as reducing and capping agent. ► Mean size of 7.5 nm, narrow size distribution, and spherical shape of particles. ► Increased sensitivity and reduced energetic dependence compared with pure alanine. ► The nanocomposite has potential application for ESR dosimetry.

Prolonged continuous intravenous infusion of the dipeptide L-alanine- L-glutamine significantly increases plasma glutamine and alanine without elevating brain glutamate in patients with severe traumatic brain injury.

Science.gov (United States)

Nägeli, Mirjam; Fasshauer, Mario; Sommerfeld, Jutta; Fendel, Angela; Brandi, Giovanna; Stover, John F

2014-07-02

Low plasma glutamine levels are associated with worse clinical outcome. Intravenous glutamine infusion dose- dependently increases plasma glutamine levels, thereby correcting hypoglutaminemia. Glutamine may be transformed to glutamate which might limit its application at a higher dose in patients with severe traumatic brain injury (TBI). To date, the optimal glutamine dose required to normalize plasma glutamine levels without increasing plasma and cerebral glutamate has not yet been defined. Changes in plasma and cerebral glutamine, alanine, and glutamate as well as indirect signs of metabolic impairment reflected by increased intracranial pressure (ICP), lactate, lactate-to-pyruvate ratio, electroencephalogram (EEG) activity were determined before, during, and after continuous intravenous infusion of 0.75 g L-alanine-L-glutamine which was given either for 24 hours (group 1, n = 6) or 5 days (group 2, n = 6) in addition to regular enteral nutrition. Lab values including nitrogen balance, urea and ammonia were determined daily. Continuous L-alanine-L-glutamine infusion significantly increased plasma and cerebral glutamine as well as alanine levels, being mostly sustained during the 5 day infusion phase (plasma glutamine: from 295 ± 62 to 500 ± 145 μmol/ l; brain glutamine: from 183 ± 188 to 549 ± 120 μmol/ l; plasma alanine: from 327 ± 91 to 622 ± 182 μmol/ l; brain alanine: from 48 ± 55 to 89 ± 129 μmol/ l; p alanine-L-glutamine infusion (0.75 g/ kg/ d up to 5 days) increased plasma and brain glutamine and alanine levels. This was not associated with elevated glutamate or signs of potential glutamate-mediated cerebral injury. The increased nitrogen load should be considered in patients with renal and hepatic dysfunction. Clinicaltrials.gov NCT02130674. Registered 5 April 2014.

Changes in alanine turnover rate due to nutritional and genetic obesity in the rat.

Science.gov (United States)

Yebras, M; Salvadó, J; Arola, L; Remesar, X; Segués, T

1994-08-01

The changes in alanine turnover were determined in Zucker rats, which were either genetically obese (fa/fa) or rendered obese by dietary treatment (cafeteria fed). The whole body rate of alanine turnover was higher in genetically obese rats than in rats in which obesity was induced by diet (cafeteria). This is possibly due to variations in the rate of the amino acid incorporation into proteins, since the rate of whole body alanine degradation is the same for both groups. Thus, the different pattern followed by alanine turnover rate in these types of obese animals reflects the differences in the nitrogen economy of these animals, pointing to a higher alanine utilization in the genetically obese animals and a conservative management of alanine in the cafeteria-fed animals.

Substrate specificity of the aspartate:alanine antiporter (AspT) of Tetragenococcus halophilus in reconstituted liposomes.

Science.gov (United States)

Sasahara, Ayako; Nanatani, Kei; Enomoto, Masaru; Kuwahara, Shigefumi; Abe, Keietsu

2011-08-19

The aspartate:alanine antiporter (AspT) of the lactic acid bacterium Tetragenococcus halophilus is a member of the aspartate:alanine exchanger (AAEx) transporter family. T. halophilus AspT catalyzes the electrogenic exchange of L-aspartate(1-) with L-alanine(0). Although physiological functions of AspT were well studied, L-aspartate(1-):L-alanine(0) antiport mechanisms are still unsolved. Here we report that the binding sites of L-aspartate and L-alanine are independently present in AspT by means of the kinetic studies. We purified His(6)-tagged T. halophilus AspT and characterized its kinetic properties when reconstituted in liposomes (K(m) = 0.35 ± 0.03 mm for L-aspartate, K(m) = 0.098 ± 0 mm for D-aspartate, K(m) = 26 ± 2 mm for L-alanine, K(m) = 3.3 ± 0.2 mm for D-alanine). Competitive inhibition by various amino acids of L-aspartate or L-alanine in self-exchange reactions revealed that L-cysteine selectively inhibited L-aspartate self-exchange but only weakly inhibited L-alanine self-exchange. Additionally, L-serine selectively inhibited L-alanine self-exchange but barely inhibited L-aspartate self-exchange. The aspartate analogs L-cysteine sulfinic acid, L-cysteic acid, and D-cysteic acid competitively and strongly inhibited L-aspartate self-exchange compared with L-alanine self-exchange. Taken together, these kinetic data suggest that the putative binding sites of L-aspartate and L-alanine are independently located in the substrate translocation pathway of AspT.

Substrate Specificity of the Aspartate:Alanine Antiporter (AspT) of Tetragenococcus halophilus in Reconstituted Liposomes*

Science.gov (United States)

Sasahara, Ayako; Nanatani, Kei; Enomoto, Masaru; Kuwahara, Shigefumi; Abe, Keietsu

2011-01-01

The aspartate:alanine antiporter (AspT) of the lactic acid bacterium Tetragenococcus halophilus is a member of the aspartate:alanine exchanger (AAEx) transporter family. T. halophilus AspT catalyzes the electrogenic exchange of l-aspartate1− with l-alanine0. Although physiological functions of AspT were well studied, l-aspartate1−:l-alanine0 antiport mechanisms are still unsolved. Here we report that the binding sites of l-aspartate and l-alanine are independently present in AspT by means of the kinetic studies. We purified His6-tagged T. halophilus AspT and characterized its kinetic properties when reconstituted in liposomes (Km = 0.35 ± 0.03 mm for l-aspartate, Km = 0.098 ± 0 mm for d-aspartate, Km = 26 ± 2 mm for l-alanine, Km = 3.3 ± 0.2 mm for d-alanine). Competitive inhibition by various amino acids of l-aspartate or l-alanine in self-exchange reactions revealed that l-cysteine selectively inhibited l-aspartate self-exchange but only weakly inhibited l-alanine self-exchange. Additionally, l-serine selectively inhibited l-alanine self-exchange but barely inhibited l-aspartate self-exchange. The aspartate analogs l-cysteine sulfinic acid, l-cysteic acid, and d-cysteic acid competitively and strongly inhibited l-aspartate self-exchange compared with l-alanine self-exchange. Taken together, these kinetic data suggest that the putative binding sites of l-aspartate and l-alanine are independently located in the substrate translocation pathway of AspT. PMID:21719707

EPR dosimetry of radiotherapy photon beams in inhomogeneous media using alanine films

Energy Technology Data Exchange (ETDEWEB)

Oesteraas, Bjoern Helge [Department of Radiation Biology, Institute for Cancer Research, Norwegian Radium Hospital, Montebello, N-0310 Oslo (Norway); Hole, Eli Olaug [Department of Physics, University of Oslo, PO Box 1048 Blindern, N-0316 Oslo (Norway); Olsen, Dag Rune [Department of Radiation Biology, Institute for Cancer Research, The Norwegian Radium Hospital, Montebello, N-0310 Oslo (Norway); Malinen, Eirik [Department of Radiation Biology, Institute for Cancer Research, The Norwegian Radium Hospital, Montebello, N-0310 Oslo (Norway)

2006-12-21

In the current work, EPR (electron paramagnetic resonance) dosimetry using alanine films (134 {mu}m thick) was utilized for dose measurements in inhomogeneous phantoms irradiated with radiotherapy photon beams. The main phantom material was PMMA, while either Styrofoam or aluminium was introduced as an inhomogeneity. The phantoms were irradiated to a maximum dose of about 30 Gy with 6 or 15 MV photons. The performance of the alanine film dosimeters was investigated and compared to results from ion chamber dosimetry, Monte Carlo simulations and radiotherapy treatment planning calculations. It was found that the alanine film dosimeters had a linear dose response above approximately 5 Gy, while a background signal obscured the response at lower dose levels. For doses between 5 and 60 Gy, the standard deviation of single alanine film dose estimates was about 2%. The alanine film dose estimates yielded results comparable to those from the Monte Carlo simulations and the ion chamber measurements, with absolute differences between estimates in the order of 1-15%. The treatment planning calculations exhibited limited applicability. The current work shows that alanine film dosimetry is a method suitable for estimating radiotherapeutical doses and for dose measurements in inhomogeneous media.

EPR dosimetry of radiotherapy photon beams in inhomogeneous media using alanine films

International Nuclear Information System (INIS)

Oesteraas, Bjoern Helge; Hole, Eli Olaug; Olsen, Dag Rune; Malinen, Eirik

2006-01-01

In the current work, EPR (electron paramagnetic resonance) dosimetry using alanine films (134 μm thick) was utilized for dose measurements in inhomogeneous phantoms irradiated with radiotherapy photon beams. The main phantom material was PMMA, while either Styrofoam or aluminium was introduced as an inhomogeneity. The phantoms were irradiated to a maximum dose of about 30 Gy with 6 or 15 MV photons. The performance of the alanine film dosimeters was investigated and compared to results from ion chamber dosimetry, Monte Carlo simulations and radiotherapy treatment planning calculations. It was found that the alanine film dosimeters had a linear dose response above approximately 5 Gy, while a background signal obscured the response at lower dose levels. For doses between 5 and 60 Gy, the standard deviation of single alanine film dose estimates was about 2%. The alanine film dose estimates yielded results comparable to those from the Monte Carlo simulations and the ion chamber measurements, with absolute differences between estimates in the order of 1-15%. The treatment planning calculations exhibited limited applicability. The current work shows that alanine film dosimetry is a method suitable for estimating radiotherapeutical doses and for dose measurements in inhomogeneous media

Structural features and kinetic characterization of alanine racemase from Bacillus pseudofirmus OF4.

Science.gov (United States)

Dong, Hui; Hu, Tingting; He, Guangzheng; Lu, Deren; Qi, Jianxun; Dou, Yanshu; Long, Wei; He, Xin; Ju, Jiansong; Su, Dan

2018-02-26

Alanine racemase (Alr) is a pyridoxal-5'-phosphate-dependent (PLP) enzyme that catalyzes a reversible racemization between the enantiomers of alanine. d-Alanine is an indispensable constituent in the biosynthesis of bacterial cell-wall peptidoglycan, and its inhibition is lethal to prokaryotes, which makes it an attractive target for designing antibacterial drugs. In this study, the molecular structure of alanine racemase from Bacillus pseudofirmus OF4 (DadX OF4 ) was determined by X-ray crystallography to a resolution of 1.8 Å. The comparison of DadX OF4 with alanine racemases from other bacteria demonstrated a conserved overall fold. Enzyme kinetics analysis showed that the conserved residues at the substrate entryway and the salt bridge at the dimer interface are critical for enzyme activity. These structural and biochemical findings provide a template for future structure-based drug-development efforts targeting alanine racemases. Copyright © 2018 Elsevier Inc. All rights reserved.

Alanine Radiation Detectors in Therapeutic Carbon Ion Beams

DEFF Research Database (Denmark)

Herrmann, Rochus; Jäkel, Oliver; Palmans, Hugo

of the depth dose curves. Solid state detectors, such as diamond detectors, radiochromic films, TLDs and the amino acid alanine are used due to there good spatial resolution. If used in particle beams their response often exhibits a dependence on particle energy and type, so the acquired signal is not always...... proportional to absorbed dose. A model by Hansen and Olsen, based on the Track Structure Theory is available, which can predict the relative efficiency of some detectors, when the particle spectrum is known. For alanine detectors the model was successfully validated by Hansen and Olsen for several ion species...... at energies below 20 MeV/u. We implemented this model in the Monte Carlo code FLUKA. At the GSI heavy ion facility in Darmstadt, Germany, alanine has been irradiated with carbon ions at energies between 88 an 400 MeV/u, which is the energy range used for therapy. The irradiation and the detector response have...

Interleukin-10 to tumor necrosis factor-alpha ratio is a predictive biomarker in nonalcoholic fatty liver disease: interleukin-10 to tumor necrosis factor-alpha ratio in steatohepatitis.

Science.gov (United States)

Hashem, Reem M; Mahmoud, Mona F; El-Moselhy, Mohamed A; Soliman, Hala M

2008-10-01

Fatty liver disease is commonly associated with diabetes mellitus (DM). Insulin resistance (IR) as an investigative biomarker is only concerned with fatty liver that results from DM type 2 associated with metabolic syndrome. Irrespective of IR, DM is generally characterized by overproduction of the proinflammatory cytokine tumor necrosis factor-alpha (TNF-alpha), whereas action of the latter is modulated by the anti-inflammatory cytokine interleukin-10 (IL-10). The aim of this study was to investigate the efficacy of using TNF-alpha alone or IL-10/TNF-alpha ratio compared to IR, as a promising biomarker for fatty liver assessment in DM. Furthermore, we hypothesized that using garlic as an immunomodulator may decrease TNF-alpha and increase IL-10 production to improve steatohepatitis. DM was induced metabolically by a high-fat diet to bring about IR, or chemically by alloxan, producing insulin deficiency, in male albino rats. Garlic powder was supplemented (15 mg/kg per day) for 3 weeks. Fatty liver was depicted histologically and biochemically (aspartic aminotransferase, alanine aminotransferase, HOMA-IR, TNF-alpha, IL-10, IL-10/TNF-alpha ratio). We found that, in contrast to obese rats, garlic decreased IL-10/TNF-alpha ratio, despite decreasing TNF-alpha in alloxan diabetic rats in agreement with the histology, which revealed more prominent improvement in the obese group. Moreover, the effect of garlic was not linked to improvement of IR in obese rats. We conclude that IL-10/TNF-alpha ratio may be considered as a convenient biomarker for investigation of fatty liver of different grades, apart from being associated with IR, and immunomodulation of this ratio in favor of increasing it may exert significant improvement.

The Discovery of the Double Helix

CERN Multimedia

CERN. Geneva

2011-01-01

Professor James D. Watson has kindly agreed to make a presentation on the 1953 finding of the Double Helix at the Cavendish Laboratory by Francis Crick and himself. Being one of the greatest scientific discoveries in human history, little else needs to be added.

A gene duplication led to specialized gamma-aminobutyrate and beta-alanine aminotransferase in yeast

DEFF Research Database (Denmark)

Andersen, Gorm; Andersen, Birgit; Dobritzsch, D.

2007-01-01

and related yeasts have two different genes/enzymes to apparently 'distinguish' between the two reactions in a single cell. It is likely that upon duplication similar to 200 million years ago, a specialized Uga1p evolved into a 'novel' transaminase enzyme with broader substrate specificity.......In humans, beta-alanine (BAL) and the neurotransmitter gamma-aminobutyrate (GABA) are transaminated by a single aminotransferase enzyme. Apparently, yeast originally also had a single enzyme, but the corresponding gene was duplicated in the Saccharomyces kluyveri lineage. SkUGA1 encodes a homologue...... to characterize the substrate specificity and kinetic parameters of the four enzymes. It was found that the cofactor pyridoxal 5'-phosphate is needed for enzymatic activity and alpha-ketoglutarate, and not pyruvate, as the amino group acceptor. SkPyd4p preferentially uses BAL as the amino group donor (V...

Green reduction of graphene oxide using alanine

International Nuclear Information System (INIS)

Wang, Jiabin; Salihi, Elif Caliskan; Šiller, Lidija

2017-01-01

There remains a real need for the easy, eco-friendly and scalable preparation method of graphene due to various potential applications. Chemical reduction is the most versatile method for the large scale production of graphene. Here we report the operating conditions for a one-step, economical and green synthesis method for the reduction of graphene oxide using a biomolecule (alanine). Graphene oxide was produced by the oxidation and exfoliation of natural graphite flake with strong oxidants using Hummers method (Hummers and Offeman, 1958), but the method was revised in our laboratory to set up a safe and environmentally friendly route. The reduction of graphene oxide was investigated using alanine at various operating conditions in order to set up optimum conditions (treatment time, temperature and concentration of the reagent). Samples have been characterized by using UV–Visible spectroscopy, Fourier transform infrared spectroscopy, transmission electron microscopy, Raman spectroscopy and X-ray diffraction analysis. - Highlights: • An environmentally friendly route was reported for the chemical reduction of graphene oxide (GO). • Alanine could reduce GO to rGO (reduced graphene oxide) without using any stabilizer or alcaline medium. • Characterization studies confirmed the successful deoxygenation of GO.

Green reduction of graphene oxide using alanine

Energy Technology Data Exchange (ETDEWEB)

Wang, Jiabin [Newcastle University, School of Chemical Engineering and Advanced Materials, Newcastle upon Tyne NE1 7RU (United Kingdom); Salihi, Elif Caliskan, E-mail: caliskanelif@gmail.com [Newcastle University, School of Chemical Engineering and Advanced Materials, Newcastle upon Tyne NE1 7RU (United Kingdom); Marmara University, Faculty of Pharmacy, Department of Basic Pharmaceutical Sciences, 34668 Istanbul (Turkey); Šiller, Lidija [Newcastle University, School of Chemical Engineering and Advanced Materials, Newcastle upon Tyne NE1 7RU (United Kingdom)

2017-03-01

There remains a real need for the easy, eco-friendly and scalable preparation method of graphene due to various potential applications. Chemical reduction is the most versatile method for the large scale production of graphene. Here we report the operating conditions for a one-step, economical and green synthesis method for the reduction of graphene oxide using a biomolecule (alanine). Graphene oxide was produced by the oxidation and exfoliation of natural graphite flake with strong oxidants using Hummers method (Hummers and Offeman, 1958), but the method was revised in our laboratory to set up a safe and environmentally friendly route. The reduction of graphene oxide was investigated using alanine at various operating conditions in order to set up optimum conditions (treatment time, temperature and concentration of the reagent). Samples have been characterized by using UV–Visible spectroscopy, Fourier transform infrared spectroscopy, transmission electron microscopy, Raman spectroscopy and X-ray diffraction analysis. - Highlights: • An environmentally friendly route was reported for the chemical reduction of graphene oxide (GO). • Alanine could reduce GO to rGO (reduced graphene oxide) without using any stabilizer or alcaline medium. • Characterization studies confirmed the successful deoxygenation of GO.

Implementation of an alanine dosimetry service

International Nuclear Information System (INIS)

Gago Arias, A.; Nunez Pelaez, N.; Peteiro Vilaseco, E.; Gomez Rodriguez, F.; Gonzalez Castano, D. M.

2011-01-01

This work facing the implementation of an alanine dosimetry service, linked to the installation of Co 6 0 Radio physics Laboratory (LP) and Paramagnetic Resonance Service of the University of Santiago de Compostela (USC).

Modulating Transmembrane α-Helix Interactions through pH-Sensitive Boundary Residues.

Science.gov (United States)

Ng, Derek P; Deber, Charles M

2016-08-09

Changes in pH can alter the structure and activity of proteins and may be used by the cell to control molecular function. This coupling can also be used in non-native applications through the design of pH-sensitive biomolecules. For example, the pH (low) insertion peptide (pHLIP) can spontaneously insert into a lipid bilayer when the pH decreases. We have previously shown that the α-helicity and helix-helix interactions of the TM2 α-helix of the proteolipid protein (PLP) are sensitive to the local hydrophobicity at its C-terminus. Given that there is an ionizable residue (Glu-88) at the C-terminus of this transmembrane (TM) segment, we hypothesized that changing the ionization state of this residue through pH may alter the local hydrophobicity of the peptide enough to affect both its secondary structure and helix-helix interactions. To examine this phenomenon, we synthesized peptide analogues of the PLP TM2 α-helix (wild-type sequence (66)AFQYVIYGTASFFFLYGALLLAEGF(90)). Using circular dichroism and Förster resonance energy transfer in the membrane-mimetic detergent sodium dodecyl sulfate, we found that a decrease in pH increases both peptide α-helicity and the extent of self-association. This pH-dependent effect is due specifically to the presence of Glu-88 at the C-terminus. Additional experiments in which Phe-90 was mutated to residues of varying hydrophobicities indicated that the strength of this effect is dependent on the local hydrophobicity near Glu-88. Our results have implications for the design of TM peptide switches and improve our understanding of how membrane protein structure and activity can be regulated through local molecular environmental changes.

Exploring the membrane fusion mechanism through force-induced disassembly of HIV-1 six-helix bundle

Energy Technology Data Exchange (ETDEWEB)

Gao, Kai [Key Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101 (China); Beijing Key Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101 (China); University of Chinese Academy of Sciences, Beijing 100049 (China); Zhang, Yong [Key Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101 (China); Beijing Key Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101 (China); Lou, Jizhong, E-mail: jlou@ibp.ac.cn [Key Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101 (China); Beijing Key Laboratory of Noncoding RNA, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101 (China)

2016-05-13

Enveloped virus, such as HIV-1, employs membrane fusion mechanism to invade into host cell. HIV-1 gp41 ectodomain uses six-helix bundle configuration to accomplish this process. Using molecular dynamic simulations, we confirmed the stability of this six-helix bundle by showing high occupancy of hydrogen bonds and hydrophobic interactions. Key residues and interactions important for the bundle integration were characterized by force-induced unfolding simulations of six-helix bundle, exhibiting the collapse order of these groups of interactions. Moreover, our results in some way concerted with a previous theory that the formation of coiled-coil choose a route which involved cooperative interactions between the N-terminal and C-terminal helix. -- Highlights: •Unfolding of HIV-1 gp41 six-helix bundle is studied by molecular dynamics simulations. •Specific interactions responsible for the stability of HIV-1 envelope post-fusion conformation were identified. •The gp41 six-helix bundle transition inducing membrane fusion might be a cooperative process of the three subunits.

Raman study of magnesium induced conversion of polyU·polyA duplexes to polyU·polyA·polyU triplexes

OpenAIRE

Herrera, S. J. Espinoza; Štepánek, J.

2010-01-01

Raman titration experiment with magnesium salt added gradually to aqueous solution of duplexes formed by RNA homopolynucleotides polyU and polyA was performed to reveal its effect on homopolynucleotide complexes. Statistical analysis of obtained spectral set has confirmed the effect already found by less structurally sensitive methods [Nucleic Acids Res. 31(17) (2003), 5101–5107] that at sufficiently high concentrations magnesium causes transformation of polyU·polyA duplexes to polyU·polyA·po...

FPGA helix tracking algorithm for PANDA

Energy Technology Data Exchange (ETDEWEB)

Liang, Yutie; Galuska, Martin; Gessler, Thomas; Kuehn, Wolfgang; Lange, Jens Soeren; Muenchow, David; Spruck, Bjoern [II. Physikalisches Institut, Giessen University (Germany); Ye, Hua [Institute of High Energy Physics, Beijing (China); Collaboration: PANDA-Collaboration

2015-07-01

The PANDA detector is a general-purpose detector for physics with high luminosity cooled antiproton beams, planed to operate at the FAIR facility in Darmstadt, Germany. The central detector includes a silicon Micro Vertex Detector (MVD) and a Straw Tube Tracker (STT). Without any hardware trigger, large amounts of raw data are streaming into the data acquisition system. The data reduction task is performed in the online system by reconstruction algorithms programmed on FPGAs (Field Programmable Gate Arrays) as first level and on a farm of GPUs or PCs as a second level. One important part in the system is the online track reconstruction. In this presentation, an online tracking algorithm for helix tracking reconstruction in the solenoidal field is shown. The tracking algorithm is composed by two parts, a road finding module followed by an iterative helix parameter calculation module. A performance study using C++ and the status of the VHDL implementation are presented.

Intersegment interactions and helix-coil transition within the generalized model of polypeptide chains approach

Science.gov (United States)

Badasyan, A. V.; Hayrapetyan, G. N.; Tonoyan, Sh. A.; Mamasakhlisov, Y. Sh.; Benight, A. S.; Morozov, V. F.

2009-09-01

The generalized model of polypeptide chains is extended to describe the helix-coil transition in a system comprised of two chains interacting side-by-side. The Hamiltonian of the model takes into account four possible types of interactions between repeated units of the two chains, i.e., helix-helix, helix-coil, coil-helix, and coil-coil. Analysis reveals when the energy Ihh+Icc of (h-h, c-c) interactions overwhelms the energy Ihc+Ich of mixed (h-c, c-h) interactions, the correlation length rises substantially, resulting in narrowing of the transition interval. In the opposite case, when Ihh+Icchelix formation and disfavored intersegment interactions from the same theoretical perspective.

Alanine-EPR as a transfer standard dosimetry system for low energy X radiation

International Nuclear Information System (INIS)

Khoury, H.J.; Silva, E.J. da; Mehta, K.; Barros, V.S. de; Asfora, V.K.; Guzzo, P.L.; Parker, A.G.

2015-01-01

The purpose of this paper is to evaluate the use of alanine-EPR as a transfer standard dosimetry system for low energy X radiation, such as that in RS-2400, which operates in the range from 25 to 150 kV and 2 to 45 mA. Two types of alanine dosimeters were investigated. One is a commercial alanine pellets from Aérial-Centre de Ressources Technologiques, France and one was prepared in our laboratory (LMRI-DEN/UFPE). The EPR spectra of the irradiated dosimeters were recorded in the Nuclear Energy Department of UFPE, using a Bruker EMX10 EPR spectrometer operating in the X-band. The alanine-EPR dosimetry system was calibrated in the range of 20–220 Gy in this X-ray field, against an ionization chamber calibrated at the relevant X-ray energy with traceability to PTB. The results showed that both alanine dosimeters presented a linear dose response the same sensitivity, when the EPR signal was normalized to alanine mass. The total uncertainty in the measured dose was estimated to be about 3%. The results indicate that it is possible to use the alanine-EPR dosimetry system for validation of a low-energy X ray irradiator, such as RS-2400.

Regulation of TCF ETS-domain transcription factors by helix-loop-helix motifs.

Science.gov (United States)

Stinson, Julie; Inoue, Toshiaki; Yates, Paula; Clancy, Anne; Norton, John D; Sharrocks, Andrew D

2003-08-15

DNA binding by the ternary complex factor (TCF) subfamily of ETS-domain transcription factors is tightly regulated by intramolecular and intermolecular interactions. The helix-loop-helix (HLH)-containing Id proteins are trans-acting negative regulators of DNA binding by the TCFs. In the TCF, SAP-2/Net/ERP, intramolecular inhibition of DNA binding is promoted by the cis-acting NID region that also contains an HLH-like motif. The NID also acts as a transcriptional repression domain. Here, we have studied the role of HLH motifs in regulating DNA binding and transcription by the TCF protein SAP-1 and how Cdk-mediated phosphorylation affects the inhibitory activity of the Id proteins towards the TCFs. We demonstrate that the NID region of SAP-1 is an autoinhibitory motif that acts to inhibit DNA binding and also functions as a transcription repression domain. This region can be functionally replaced by fusion of Id proteins to SAP-1, whereby the Id moiety then acts to repress DNA binding in cis. Phosphorylation of the Ids by cyclin-Cdk complexes results in reduction in protein-protein interactions between the Ids and TCFs and relief of their DNA-binding inhibitory activity. In revealing distinct mechanisms through which HLH motifs modulate the activity of TCFs, our results therefore provide further insight into the role of HLH motifs in regulating TCF function and how the inhibitory properties of the trans-acting Id HLH proteins are themselves regulated by phosphorylation.

Selective intercalation of six ligands molecules in a self-assembled triple helix

NARCIS (Netherlands)

Mateos timoneda, Miguel; Kerckhoffs, J.M.C.A.; Reinhoudt, David; Crego Calama, Mercedes

2007-01-01

The addition of a ligand molecule to an artificial self-assembled triple helix leads to the selective intercalation of two hydrogen-bonded trimers in specific binding pockets. Furthermore, the triple helix suffers large conformational rearrangements in order to accommodate the ligand molecules in a

Copper(II) 12-metallacrown-4 complexes of alpha-, beta- and gamma-aminohydroxamic acids: a comparative thermodynamic study in aqueous solution.

Science.gov (United States)

Tegoni, Matteo; Remelli, Maurizio; Bacco, Dimitri; Marchiò, Luciano; Dallavalle, Francesco

2008-05-28

A complete thermodynamic study of the protonation and Cu(II) complex formation equilibria of a series of alpha- and beta-aminohydroxamic acids in aqueous solution was performed. The thermodynamic parameters obtained for the protonation of glycine-, (S)-alpha-alanine-, (R,S)-valine-, (S)-leucine-, beta-alanine- and (R)-aspartic-beta-hydroxamic acids were compared with those previously reported for gamma-amino- and (S)-glutamic-gamma-hydroxamic acids. The enthalpy/entropy parameters calculated for the protonation microequilibria of these three types of ligands are in very good agreement with the literature values for simple amines and hydroxamic acids. The pentanuclear complexes [Cu5L4H(-4)]2+ contain the ligands acting as (NH2,N-)-(O,O-) bridging bis-chelating and correspond to 12-metallacrown-4 (12-MC-4) which are formed by self-assembly between pH 4 and 6 with alpha-aminohydroxamates (HL), while those with beta- and gamma-derivatives exist in a wider pH range (4-11). The stability order of these metallomacrocycles is beta- > alpha- > gamma-aminohydroxamates. The formation of 12-MC-4 with alpha-aminohydroxamates is entropy-driven, and that with beta-derivatives is enthalpy-driven, while with gamma-GABAhydroxamate both effects occur. These results are interpreted on the basis of specific enthalpies or entropy contributions related to chelate ring dimensions, charge neutralization and solvation-desolvation effects. The enthalpy/entropy parameters of 12-MC-4 with alpha-aminohydroxamic acids considered are also dependent on the optical purity of the ligands. Actually, that with (R,S)-valinehydroxamic acid presents an higher entropy and a lower enthalpy value than those of enantiopure ligands, although the corresponding stabilities are almost equivalent. Moreover, DFT calculations are in agreement with a more exothermic enthalpy found for metallacrowns with enantiomerically pure ligands.

Thermal helix-coil transition in UV irradiated collagen from rat tail tendon.

Science.gov (United States)

Sionkowska, A; Kamińska, A

1999-05-01

The thermal helix-coil transition in UV irradiated collagen solution, collagen film and pieces of rat tail tendon (RTT) were compared. Their thermal stability's were determined by differential scanning calorimeter (DSC) and by viscometric measurements. The denaturation temperatures of collagen solution, film and pieces of RTT were different. The helix-coil transition occur near 40 degrees C in collagen solution, near 112 degrees C in collagen film, and near 101 degrees C in pieces of RTT. After UV irradiation the thermal helix-coil transition of collagen samples were changed. These changes depend on the degree of hydratation.

Effect of beta-alanine, with and without sodium bicarbonate, on 2000-m rowing performance.

Science.gov (United States)

Hobson, Ruth M; Harris, Roger C; Martin, Dan; Smith, Perry; Macklin, Ben; Gualano, Bruno; Sale, Craig

2013-10-01

To examine the effect of beta-alanine only and beta-alanine with sodium bicarbonate supplementation on 2,000-m rowing performance. Twenty well-trained rowers (age 23 ± 4 y; height 1.85 ± 0.08 m; body mass 82.5 ± 8.9 kg) were assigned to either a placebo or beta-alanine (6.4 g · d(-1) for 4 weeks) group. A 2,000-m rowing time trial (TT) was performed before supplementation (Baseline) and after 28 and 30 days of supplementation. The post supplementation trials involved supplementation with either maltodextrin or sodium bicarbonate in a double-blind, crossover design, creating four study conditions (placebo with maltodextrin; placebo with sodium bicarbonate; beta-alanine with maltodextrin; beta-alanine with sodium bicarbonate). Blood lactate, pH, bicarbonate, and base excess were measured pre-TT, immediately post-TT and at TT+5 min. Performance data were analyzed using magnitude based inferences. Beta-alanine supplementation was very likely to be beneficial to 2,000-m rowing performance (6.4 ± 8.1 s effect compared with placebo), with the effect of sodium bicarbonate having a likely benefit (3.2 ± 8.8 s). There was a small (1.1 ± 5.6 s) but possibly beneficial additional effect when combining chronic beta-alanine supplementation with acute sodium bicarbonate supplementation compared with chronic beta-alanine supplementation alone. Sodium bicarbonate ingestion led to increases in plasma pH, base excess, bicarbonate, and lactate concentrations. Both chronic beta-alanine and acute sodium bicarbonate supplementation alone had positive effects on 2,000-m rowing performance. The addition of acute sodium bicarbonate to chronic beta-alanine supplementation may further enhance rowing performance.

L-alanine uptake in membrane vesicles from Mytilus edulis gills

International Nuclear Information System (INIS)

Pajor, A.M.; Wright, S.H.

1986-01-01

Previous studies have shown that gills from M. edulis can accumulate L-alanine from seawater by a saturable process specific for α-neutral amino acids. This uptake occurs against chemical gradients in excess of 10 6 to 1. To further characterize this uptake, membrane vesicles were prepared from M. edulis gill tissue by differential centrifugation. Enrichments of putative enzyme markers (relative to that in combined initial fractions) were as follows: γ-Glutamyltranspeptidase, 25-30x; Alkaline Phosphatase, 5-6x; K + -dependent para-Nitrophenyl Phosphatase, 3-5x; Succinate Dehydrogenase 0.1-0.2x. These results suggest that the preparation is enriched in plasma membranes, although histochemical studies will be needed to verify this. The time course of 14 C-L-alanine uptake in the presence of inwardly-directed Na + gradient showed a transient overshoot (3-5 fold) at 10 minutes which decreased to equilibrium after six hours. The size of the overshoot and early uptake rates depended on the size of the inwardly-directed Na + gradient. No overshoot was seen in the presence of inwardly-directed gradients of LiCl or choline-Cl, or with equilibrium concentrations NaCl or mannitol. A reduced overshoot was seen with a gradient of NaSCN. A small overshoot was seen with an inwardly-directed gradient of KCl. Transport of L-alanine included saturable and diffusive components. Uptake of 6 μM L-alanine was inhibited more than 80% by 100 μM α-zwitterionic amino acids (alanine, leucine, glycine); by 30 to 75% by proline, aspartate and lysine; and less than 20% by a β-amino acid, taurine. The results of these experiments agree with those from intact gill studies and support the hypothesis that L-alanine is transported into gill epithelial cells by a secondary active transport process involving Na +

Spectrophotometric readout for an alanine dosimeter for food irradiation applications

International Nuclear Information System (INIS)

Ebraheem, S.; Beshir, W.B.; Eid, S.; Sobhy, R.; Kovacs, A.

2003-01-01

The alanine-electron spin resonance (EPR) readout system is well known as a reference and transfer dosimetry system for the evaluation of high doses in radiation processing. The high cost of an EPR/alanine dosimetry system is a serious handicap for large-scale routine application in irradiation facilities. In this study, the use of a complex produced by dissolving irradiated L-alanine in 1,4-phenyl diammonium dichloride solution was investigated for dosimetry purposes. This complex--having a purple colour--has an increasing absorbance with increasing dose in the range of 1-20 kGy. The applicability of spectrophotometric evaluation was studied by measuring the absorbance intensity of this complex at 360 and 505 nm, respectively. Fluorimetric evaluation was also investigated by measuring the emission of the complex at 435 nm as a function of dose. The present method is easy for routine application. The effect of the dye concentration as well as the suitable amount of irradiated alanine has been studied. With respect to routine application, the stability of the product complex after its formation was also investigated

Triple helix interactions for eco-innovation

DEFF Research Database (Denmark)

Hermann, Roberto Rivas; Riisgaard, Henrik; Remmen, Arne

the role of science parks in promoting eco-innovation. This study uses qualitative data gathered in two units of analysis: Panama Canal Authority and City of Knowledge Science Park. The study examines how Triple Helix interactions have built the regional system of eco-innovation at the Panama Canal...

PGC-1α regulates alanine metabolism in muscle cells.

Science.gov (United States)

Hatazawa, Yukino; Qian, Kun; Gong, Da-Wei; Kamei, Yasutomi

2018-01-01

The skeletal muscle is the largest organ in the human body, depositing energy as protein/amino acids, which are degraded in catabolic conditions such as fasting. Alanine is synthesized and secreted from the skeletal muscle that is used as substrates of gluconeogenesis in the liver. During fasting, the expression of PGC-1α, a transcriptional coactivator of nuclear receptors, is increased in the liver and regulates gluconeogenesis. In the present study, we observed increased mRNA expression of PGC-1α and alanine aminotransferase 2 (ALT2) in the skeletal muscle during fasting. In C2C12 myoblast cells overexpressing PGC-1α, ALT2 expression was increased concomitant with an increased alanine level in the cells and medium. In addition, PGC-1α, along with nuclear receptor ERR, dose-dependently enhanced the ALT2 promoter activity in reporter assay using C2C12 cells. In the absence of glucose in the culture medium, mRNA levels of PGC-1α and ALT2 increased. Endogenous PGC-1α knockdown in C2C12 cells reduced ALT2 gene expression level, induced by the no-glucose medium. Taken together, in the skeletal muscle, PGC-1α activates ALT2 gene expression, and alanine production may play roles in adaptation to fasting.

Application of an alanine dosimetry system for industrial irradiation and radiation protection

International Nuclear Information System (INIS)

Gohs, U.

1996-01-01

This paper reports the application of alanine dosimetry in radiation processing. Continuous checks of the EPR measuring conditions as well as using high-quality alanine dosimeters and consistent technique for dose determination guarantee an accuracy of about ± 3% intermediate dose levels. The alanine dosimetry system was applied for dose mapping measurements during irradiator qualification and performance qualification of different products, routine dosimetry, and special radiation protection applications within the gamma irradiator. (author)

Contribution of liver and skeletal muscle to alanine and lactate metabolism in humans

International Nuclear Information System (INIS)

Consoli, A.; Nurjhan, N.; Reilly, J.J. Jr.; Bier, D.M.; Gerich, J.E.

1990-01-01

To quantitate alanine and lactate gluconeogenesis in postabsorptive humans and to test the hypothesis that muscle is the principal source of these precursors, we infused normal volunteers with [3-14C]lactate, [3-13C]alanine, and [6-3H]glucose and calculated alanine and lactate incorporation into plasma glucose corrected for tricarboxylic acid cycle carbon exchange, the systemic appearance of these substrates, and their forearm fractional extraction, uptake, and release. Forearm alanine and lactate fractional extraction averaged 37 +/- 3 and 27 +/- 2%, respectively; muscle alanine release (2.94 +/- 0.27 mumol.kg body wt-1.min-1) accounted for approximately 70% of its systemic appearance (4.18 +/- 0.31 mumol.kg body wt-1.min-1); muscle lactate release (5.51 +/- 0.42 mumol.kg body wt-1.min-1) accounted for approximately 40% of its systemic appearance (12.66 +/- 0.77 mumol.kg body wt-1.min-1); muscle alanine and lactate uptake (1.60 +/- 0.7 and 3.29 +/- 0.36 mumol.kg body wt-1.min-1, respectively) accounted for approximately 30% of their overall disappearance from plasma, whereas alanine and lactate incorporation into plasma glucose (1.83 +/- 0.20 and 4.24 +/- 0.44 mumol.kg body wt-1.min-1, respectively) accounted for approximately 50% of their disappearance from plasma. We therefore conclude that muscle is the major source of plasma alanine and lactate in postabsorptive humans and that factors regulating their release from muscle may thus exert an important influence on hepatic gluconeogenesis

Democracy and environment as references for quadruple and quintuple helix innovation systems

Science.gov (United States)

Carayannis, Elias G.; Campbell, David F. J.; Orr, Barron J.

2015-04-01

The perspective of democracy and the ecological context define key references for knowledge production and innovation in innovation systems. Particularly under conditions of environmental change where enhancing the potential for adaptation is critical, this requires a closer look at ecological responsibility and sensitivity in the different innovation models and governance regimes. The "Quintuple Helix" innovation model is an approach that stresses the necessary socio-ecological transition of society and economy by adding an environment helix to an innovation system already made up of three (university-industry-government) or four (civil society relations) helices in a way that supports adaptation by incorporating global warming as both a challenge to and a driver of innovation. There is the proposition that knowledge production and innovation co-evolve with democracy (Carayannis and Campbell, 2014). In the Triple Helix model (Etzkowitz and Leydesdorff, 2000) the existence of a democracy does not appear to be necessary for knowledge production and innovation. However, the Quadruple Helix (Carayannis and Campbell, 2009, 2010 and 2014) is defined and represented by additional key attributes and components: "media-based and culture-based public", "civil society" and "arts, artistic research and arts-based innovation" (Bast, Carayannis and Campbell, 2015). Implications of this are that the fourth helix in the Quadruple Helix innovation systems brings in and represents the perspective of "dimension of democracy" or the "context of democracy" for knowledge in general and knowledge production and innovation in more particular. Within theories of democracy there is a competition between narrow and broader concepts of democracy (Campbell, 2013). This is particularly true when democracy is to be understood to transcend more substantially the narrow understanding of being primarily based on or being primarily rooted in government institutions (within a Triple Helix

Structure of bacteriophage [phi]29 head fibers has a supercoiled triple repeating helix-turn-helix motif

Energy Technology Data Exchange (ETDEWEB)

Xiang, Ye; Rossmann, Michael G. (Purdue)

2011-12-22

The tailed bacteriophage {phi}29 capsid is decorated with 55 fibers attached to quasi-3-fold symmetry positions. Each fiber is a homotrimer of gene product 8.5 (gp8.5) and consists of two major structural parts, a pseudohexagonal base and a protruding fibrous portion that is about 110 {angstrom} in length. The crystal structure of the C-terminal fibrous portion (residues 112-280) has been determined to a resolution of 1.6 {angstrom}. The structure is about 150 {angstrom} long and shows three distinct structural domains designated as head, neck, and stem. The stem region is a unique three-stranded helix-turn-helix supercoil that has not previously been described. When fitted into a cryoelectron microscope reconstruction of the virus, the head structure corresponded to a disconnected density at the distal end of the fiber and the neck structure was located in weak density connecting it to the fiber. Thin section studies of Bacillus subtilis cells infected with fibered or fiberless {phi}29 suggest that the fibers might enhance the attachment of the virions onto the host cell wall.

Structural and functional aspects of winged-helix domains at the core of transcription initiation complexes.

Science.gov (United States)

Teichmann, Martin; Dumay-Odelot, Hélène; Fribourg, Sébastien

2012-01-01

The winged helix (WH) domain is found in core components of transcription systems in eukaryotes and prokaryotes. It represents a sub-class of the helix-turn-helix motif. The WH domain participates in establishing protein-DNA and protein-protein-interactions. Here, we discuss possible explanations for the enrichment of this motif in transcription systems.

Earthworms accumulate alanine in response to drought.

Science.gov (United States)

Holmstrup, Martin; Slotsbo, Stine; Henriksen, Per G; Bayley, Mark

2016-09-01

Earthworms have ecologically significant functions in tropical and temperate ecosystems and it is therefore important to understand how these animals survive during drought. In order to explore the physiological responses to dry conditions, we simulated a natural drought incident in a laboratory trial exposing worms in slowly drying soil for about one month, and then analyzed the whole-body contents of free amino acids (FAAs). We investigated three species forming estivation chambers when soils dry out (Aporrectodea tuberculata, Aporrectodea icterica and Aporrectodea longa) and one species that does not estivate during drought (Lumbricus rubellus). Worms subjected to drought conditions (alanine that was significantly upregulated in all tested species. Alanine was the most important FAA reaching 250-650μmolg(-1) dry weight in dehydrated Aporrectodea species and 300μmolg(-1) dry weight in L. rubellus. Proline was only weakly upregulated in some species as were a few other FAAs. Species forming estivation chambers (Aporrectodea spp.) did not show a better ability to conserve body water than the non-estivating species (L. rubellus) at the same drought level. These results suggest that the accumulation of alanine is an important adaptive trait in drought tolerance of earthworms in general. Copyright © 2016 Elsevier Inc. All rights reserved.

Characteristics and application of alanine dosimeter 'Aminogray'

International Nuclear Information System (INIS)

Kashiwazaki, Shigeru; Matsuyama, Shigeki; Hatta, Toshimasa; Yagyu, Hideki; Kojima, Takuji; Tanaka, Ryuichi; Morita, Yohsuke.

1988-01-01

Recently, accompanying the progress of nuclear power generation and space development, the evaluation of reliability for the materials and parts used under irradiation has become important. For the evaluation of reliability, the accurate grasp of radiation dose is the prerequisite. In some case, the measurement of cumulative dose in a long period in an actual environment becomes necessary. In this paper, the characteristics and application of a new dosimeter element 'Aminogray' which is suitable to the above requirement are reported. Aminogray is rodshape element made by forming alanine, a kind of amino acid, using a binder polymer, and the alanine content is 70 wt.%, and the polymer is polystyrene. An element of 3 mm diameter and 30 mm length is enclosed in a polystyrene cylinder of 4 mm thickness. This thickness was determined by considering the electronic equilibrium condition in Co-60 gamma-ray irradiation. The principle of the measurement is to determine a dose by measuring the amount of free radicals produced in alanine by radiation using ESR method. The free radicals are extremely stable, and exist for a long period, and the amount of radical production is proportional to absorbed dose. The development, characteristics and application of Aminogray are reported. (K.I.)

Regulation of the ald Gene Encoding Alanine Dehydrogenase by AldR in Mycobacterium smegmatis

Science.gov (United States)

Jeong, Ji-A; Baek, Eun-Young; Kim, Si Wouk; Choi, Jong-Soon

2013-01-01

The regulatory gene aldR was identified 95 bp upstream of the ald gene encoding l-alanine dehydrogenase in Mycobacterium smegmatis. The AldR protein shows sequence similarity to the regulatory proteins of the Lrp/AsnC family. Using an aldR deletion mutant, we demonstrated that AldR serves as both activator and repressor for the regulation of ald gene expression, depending on the presence or absence of l-alanine. The purified AldR protein exists as a homodimer in the absence of l-alanine, while it adopts the quaternary structure of a homohexamer in the presence of l-alanine. The binding affinity of AldR for the ald control region was shown to be increased significantly by l-alanine. Two AldR binding sites (O1 and O2) with the consensus sequence GA-N2-ATC-N2-TC and one putative AldR binding site with the sequence GA-N2-GTT-N2-TC were identified upstream of the ald gene. Alanine and cysteine were demonstrated to be the effector molecules directly involved in the induction of ald expression. The cellular level of l-alanine was shown to be increased in M. smegmatis cells grown under hypoxic conditions, and the hypoxic induction of ald expression appears to be mediated by AldR, which senses the intracellular level of alanine. PMID:23749971

No effect of β-alanine on muscle function and kayak performance

DEFF Research Database (Denmark)

Bech, Signe Refsgaard; Nielsen, Tobias Schmidt; Hald, Martin

2018-01-01

PURPOSE: It was investigated if β-alanine supplementation counteracts muscular fatigue development or improves athletic performance. METHODS: Elite kayak rowers (10 males and 7 females) were supplemented with either 80 mg/kg body mass/day β-alanine or placebo for 8 weeks. Muscular fatigue...

Effect of β-alanine supplementation on 20 km cycling time trial performance

Directory of Open Access Journals (Sweden)

Ruth Margaret JAMES

2014-09-01

Full Text Available The effects of β-alanine supplementation on high-intensity cycling performance and capacity have been evaluated, although the effects on longer duration cycling performance are unclear. Nineteen UK category 1 male cyclists completed four 20 km cycling time trials, two before and two after supplementation with either 6.4 g•d-1 β-alanine (n = 10; BA or a matched placebo (n = 9; P. Performance time for the 20 km time trial and 1 km split times were recorded. There was no significant effect of β-alanine supplementation on 20 km time trial performance (BA-pre 1943 ± 129 s; BA-post 1950 ± 147 s; P-pre 1989 ± 106 s; P-post 1986 ± 115 s or on the performance of each 1 km split. The effect of β-alanine on 20 km time trial performance was deemed unclear as determined by magnitude based inferences. Supplementation with 6.4 g•d-1 of β-alanine for 4 weeks did not affect 20 km cycling time trial performance in well trained male cyclists.

Alanine Counteracts the Destabilizing Effect that Urea has on RNase-A.

Science.gov (United States)

Chowhan, Rimpy K; Ali, Fasil; Bhat, Mohd Y; Rahman, Safikur; Singh, Laishram R; Ahmad, Faizan; Dar, Tanveer A

2016-01-01

It is generally believed that organisms use and accumulate methylamine osmolytes to prevent urea's damaging effect on protein stability and activity. However, urea-rich cells not only accumulate methylamines but also many other methylated and non-methylated compounds as well. But, so far it is not known whether osmolytes that are not accumulated in urea-rich cells could also confer urea-counteracting properties. We investigated the behavior of a non-methylamine osmolyte, alanine for its counteracting effect against urea denaturation of a model protein, ribonuclease A (RNase-A). We have measured structure and thermodynamic parameters (Tm, ΔHm, and ΔGD°) of RNase-A in the presence of alanine, urea and their combination. The results were also compared with the ability of glycine (osmolyte lacking one methyl group when compared with alanine) to counter urea's effect on protein stability. We observed that alanine but not glycine counteracts urea's harmful effect on RNase-A stability. The results indicated that alanine (in addition to methylamine osmolytes) may serve as an alternate urea-counteractant. Since glycine fails to protect RNase-A from urea's destabilizing effect, it seems that methylation to glycine might have some evolutionary significance to protect proteins against harmful effects of urea.

Inactivation of colicin Y by intramembrane helix–helix interaction with its immunity protein

Czech Academy of Sciences Publication Activity Database

Šmajs, D.; Doležalová, M.; Macek, Pavel; Žídek, L.

2008-01-01

Roč. 275, č. 21 (2008), s. 5325-5331 ISSN 1742-464X Institutional research plan: CEZ:AV0Z50200510 Keywords : colicin immunity * colicin y * helix-helix interaction Subject RIV: CE - Biochemistry Impact factor: 3.139, year: 2008

ESR/Alanine {gamma}-dosimetry in the 10-30 Gy range

Energy Technology Data Exchange (ETDEWEB)

Fainstein, C. E-mail: cfainstein@cab.cnea.gov.ar; Winkler, E.; Saravi, M

2000-05-15

We report Alanine Dosimeter preparation, procedures for using the ESR/Dosimetry method, and the resulting calibration curve for {gamma}-irradiation in the range from 10-30 Gy. We use calibration curve to measure the irradiation dose in {gamma}-irradiation of human blood, as required in Blood Transfusion Therapy. The ESR/Alanine results are compared against those obtained using the thermoluminescent dosimetry (TLD) method.

Inducible l-Alanine Exporter Encoded by the Novel Gene ygaW (alaE) in Escherichia coli ▿

Science.gov (United States)

Hori, Hatsuhiro; Yoneyama, Hiroshi; Tobe, Ryuta; Ando, Tasuke; Isogai, Emiko; Katsumata, Ryoichi

2011-01-01

We previously isolated a mutant hypersensitive to l-alanyl-l-alanine from a non-l-alanine-metabolizing Escherichia coli strain and found that it lacked an inducible l-alanine export system. Consequently, this mutant showed a significant accumulation of intracellular l-alanine and a reduction in the l-alanine export rate compared to the parent strain. When the mutant was used as a host to clone a gene(s) that complements the dipeptide-hypersensitive phenotype, two uncharacterized genes, ygaW and ytfF, and two characterized genes, yddG and yeaS, were identified. Overexpression of each gene in the mutant resulted in a decrease in the intracellular l-alanine level and enhancement of the l-alanine export rate in the presence of the dipeptide, suggesting that their products function as exporters of l-alanine. Since ygaW exhibited the most striking impact on both the intra- and the extracellular l-alanine levels among the four genes identified, we disrupted the ygaW gene in the non-l-alanine-metabolizing strain. The resulting isogenic mutant showed the same intra- and extracellular l-alanine levels as observed in the dipeptide-hypersensitive mutant obtained by chemical mutagenesis. When each gene was overexpressed in the wild-type strain, which does not intrinsically excrete alanine, only the ygaW gene conferred on the cells the ability to excrete alanine. In addition, expression of the ygaW gene was induced in the presence of the dipeptide. On the basis of these results, we concluded that YgaW is likely to be the physiologically most relevant exporter for l-alanine in E. coli and proposed that the gene be redesignated alaE for alanine export. PMID:21531828

PH4 of petunia is an R2R3-MYB protein that activates vacuolar acidification through interactions with Basic-Helix-Loop-Helix transcription factors of the anthocyanin pathway.

NARCIS (Netherlands)

Quattrocchio, F.M.; Verweij, C.W.; Spelt, C.E.; Mol, J.N.M.; Koes, R.E.

2007-01-01

The Petunia hybrids genes ANTHOCYANIN1 (AN1) and AN2 encode transcription factors with a basic-helix-loop-helix (BHLH) and a MYB domain, respectively, that are required for anthocyanin synthesis and acidification of the vacuole in petal cells. Mutation of PH4 results in a bluer flower color,

New Generation of self-calibrated SS/EPR dosimeters: Alanine/EPR dosimeters

International Nuclear Information System (INIS)

Yordanov, N.D.; Gancheva, V.

1999-01-01

A new type of solid state/EPR dosimeters is described. Principally, it contains radiation sensitive diamagnetic material, some quantity of EPR active, but radiation insensitive, substance (for example Mn 2+ /MgO) and a binding material. In the present case alanine is used as a radiation sensitive substance. With this dosimeter, the EPR spectra of alanine and Mn 2+ are simultaneously recorded and the calibration graph represents the ratio of alanine versus Mn 2+ EPR signal intensity as a function of absorbed dose. In this way the reproducibility of the results is expected to be improved significantly including their intercomparison among different laboratories. Homogeneity of the prepared dosimeters and their behaviour (fading of EPR signals with time, influence of different meteorological conditions) show satisfactory reproducibility and stability with time. Because two different EPR active samples are recorded simultaneously, the influence of some instrument setting parameters (microwave power, modulation amplitude and modulation frequency) on the ratio I alanine /I Mn is also investigated. (author)

Effect of combined β-alanine and sodium bicarbonate supplementation on cycling performance.

Science.gov (United States)

Bellinger, Phillip M; Howe, Samuel T; Shing, Cecilia M; Fell, James W

2012-08-01

The purpose of this study was to investigate the effects of 28 d of β-alanine supplementation on 4-min cycling time trial performance and to determine whether there was an additive effect of combined β-alanine and sodium bicarbonate (NaHCO3) supplementation on high-intensity cycling performance. Fourteen highly trained cyclists (mean ± SD: age = 25.4 ± 7.2 yr, mass = 71.1 ± 7.1 kg, V˙O(2max) = 66.6 ± 5.7 mL·kg·min) supplemented for 28 d with β-alanine (65 mg·kg body mass each day) or placebo. A maximal 4-min bout of cycling was performed before supplementation (baseline) and twice after supplementation: after ingestion of NaHCO3 (300 mg·kg body mass) and ingestion of a placebo using a randomized crossover design with 2 d between trials. Blood pH and HCO3 concentration were determined before loading (postsupplementation trials) and at pretest and posttest. In the acute NaHCO3 loading trials, blood pH and HCO3 were elevated from before loading to pretest, and the magnitude of the change in HCO3 from pretest to posttest was significantly greater compared with the acute placebo loading trial (P < 0.001). Average power output in the 4-min cycling performance trial was increased in placebo + NaHCO3 (+3.1% ± 1.8%) and β-alanine + NaHCO3 (+3.3% ± 3.0%) compared with baseline (P < 0.05). β-alanine + placebo did not significantly improve average power output compared with baseline (+1.6% ± 1.7%, P = 0.20); however, magnitude-based inferences demonstrated that β-alanine + placebo was associated with a 37% likelihood of producing average power improvements. In trained cyclists, β-alanine supplementation did not significantly improve 4-min cycling performance; however, there may be a small meaningful improvement in performance. Acute NaHCO3 supplementation significantly improved 4-min cycling performance. There seemed to be a minimal additive effect of combined β-alanine and NaHCO3 supplementation.

SU-E-T-643: Pure Alanine Dosimeter for Verification Dosimetry in IMRT

International Nuclear Information System (INIS)

Al-Karmi, Anan M.; Zraiqat, Fadi

2015-01-01

Purpose: The objective of this study was evaluation of accuracy of pure alanine dosimeters measuring intensity-modulated radiation therapy (IMRT) dose distributions in a thorax phantom. Methods: Alanine dosimeters were prepared in the form of 110 mg pure L-α-alanine powder filled into clear tissue-equivalent polymethylmethacrylate (PMMA) plastic tubes with the dimensions 25 mm length, 3 mm inner diameter, and 1 mm wall thickness. A dose-response calibration curve was established for the alanine by placing the dosimeters at 1.5 cm depth in a 30×30×30 cm 3 solid water phantom and then irradiating on a linac with 6 MV photon beam at 10×10 cm 2 field size to doses ranging from 1 to 5 Gy. Electron paramagnetic resonance (EPR) spectroscopy was used to determine the absorbed dose in alanine. An IMRT treatment plan was designed for a commercial heterogeneous CIRS thorax phantom and the dose values were calculated at three different points located in tissue, lung, and bone equivalent materials. A set of dose measurements was carried out to compare measured and calculated dose values by placing the alanine dosimeters at those selected locations inside the thorax phantom and delivering the IMRT to the phantom. Results: The alanine dose measurements and the IMRT plan dose calculations were found to be in agreement within ±2%. Specifically, the deviations were −0.5%, 1.3%, and −1.7% for tissue, lung, and bone; respectively. The slightly large deviations observed for lung and bone may be attributed to tissue inhomogeneity, steep dose gradients in these regions, and uncontrollable changes in spectrometer conditions. Conclusion: The results described herein confirmed that pure alanine dosimeter was suitable for in-phantom dosimetry of IMRT beams because of its high sensitivity and acceptable accuracy. This makes the dosimeter a promising option for quality control of the therapeutic beams, complementing the commonly used ionization chambers, TLDs, and films

Chondrogenic potential of macroporous biodegradable cryogels based on synthetic poly(.alpha.-amino acids)

Czech Academy of Sciences Publication Activity Database

Sedlačík, Tomáš; Acar, O. K.; Studenovská, Hana; Kotelnikov, Ilya; Kučka, Jan; Konečná, Z.; Zikmund, T.; Kaiser, J.; Köse, G. T.; Rypáček, František

2018-01-01

Roč. 14, č. 2 (2018), s. 228-238 ISSN 1744-683X Institutional support: RVO:61389013 Keywords : poly( amino acids ) * cryogelation * scaffold Subject RIV: CD - Macromolecular Chemistry OBOR OECD: Polymer science Impact factor: 3.889, year: 2016

The Alanine Racemase of Mycobacterium smegmatis Is Essential for Growth in the Absence of d-Alanine▿ †

Science.gov (United States)

Milligan, Daniel L.; Tran, Sieu L.; Strych, Ulrich; Cook, Gregory M.; Krause, Kurt L.

2007-01-01

Alanine racemase, encoded by the gene alr, is an important enzyme in the synthesis of d-alanine for peptidoglycan biosynthesis. Strains of Mycobacterium smegmatis with a deletion mutation of the alr gene were found to require d-alanine for growth in both rich and minimal media. This indicates that alanine racemase is the only source of d-alanine for cell wall biosynthesis in M. smegmatis and confirms alanine racemase as a viable target gene for antimycobacterial drug development. PMID:17827284

Dextran Sulfate Sodium Inhibits Alanine Synthesis in Caco-2 Cells

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Carolyn M. Slupsky

2011-04-01

Full Text Available To understand and characterize the pathogenic mechanisms of inflammatory bowel disease, dextran sulfate sodium (DSS has been used to induce acute and chronic colitis in animal models by causing intestinal epithelium damage. The mechanism of action of DSS in producing this outcome is not well understood. In an effort to understand how DSS might impact epithelial cell metabolism, we studied the intestinal epithelial cell line Caco-2 incubated with 1% DSS over 56 hours using 1H NMR spectroscopy. We observed no difference in cell viability as compared to control cultures, and an approximately 1.5-fold increase in IL-6 production upon incubation with 1% DSS. The effect on Caco-2 cell metabolism as measured through changes in the concentration of metabolites in the cell supernatant included a three-fold decrease in the concentration of alanine. Given that the concentrations of other amino acids in the cell culture supernatant were not different between treated and control cultures over 56 hours suggest that DSS inhibits alanine synthesis, specifically alanine aminotransferase, without affecting other key metabolic pathways. The importance of alanine aminotransferase in inflammatory bowel disease is discussed.

Synthesis, characterization, and biocompatible properties of alanine-grafted chitosan copolymers.

Science.gov (United States)

Park, Gyu Han; Kang, Min-Sil; Knowles, Jonathan C; Gong, Myoung-Seon

2016-04-01

In order to overcome major problems regarding the lack of affinity to solvents and limited reactivity of the free amines of chitosan, introduction of appropriate spacer arms having terminal amine function is considered of interest. L-Alanine-N-carboxyanhydride was grafted onto chitosan via anionic ring-opening polymerization. The chemical and structural characterizations of L-alanine-grafted chitosan (Ala-g-Cts) were confirmed through Fourier transform infrared spectroscopy and proton nuclear magnetic resonance spectroscopy ((1)H NMR). In addition, the viscoelastic properties of Ala-g-Cts were examined by means of a rotational viscometer, and thermal analysis was carried out with a thermogravimetric analyzer and differential scanning calorimetry. Morphological changes in the chitosan L-alanine moiety were determined by x-ray diffraction. To determine the feasibility of using these films as biomedical materials, we investigated the effects of their L-alanine content on physical and mechanical properties. The biodegradation results of crosslinked Ala-g-Cts films were evaluated in phosphate-buffered solution containing lysozyme at 37℃. Proliferation of MC3T3-E1 cells on crosslinked Ala-g-Cts films was also investigated with use of the CCK-8 assay. © The Author(s) 2016.

High-Temperature unfolding of a trp-Cage mini-protein: a molecular dynamics simulation study

Directory of Open Access Journals (Sweden)

Seshasayee Aswin Sai Narain

2005-03-01

Full Text Available Abstract Background Trp cage is a recently-constructed fast-folding miniprotein. It consists of a short helix, a 3,10 helix and a C-terminal poly-proline that packs against a Trp in the alpha helix. It is known to fold within 4 ns. Results High-temperature unfolding molecular dynamics simulations of the Trp cage miniprotein have been carried out in explicit water using the OPLS-AA force-field incorporated in the program GROMACS. The radius of gyration (Rg and Root Mean Square Deviation (RMSD have been used as order parameters to follow the unfolding process. Distributions of Rg were used to identify ensembles. Conclusion Three ensembles could be identified. While the native-state ensemble shows an Rg distribution that is slightly skewed, the second ensemble, which is presumably the Transition State Ensemble (TSE, shows an excellent fit. The denatured ensemble shows large fluctuations, but a Gaussian curve could be fitted. This means that the unfolding process is two-state. Representative structures from each of these ensembles are presented here.

Lysine as helix C-capping residue in a synthetic peptide.

Science.gov (United States)

Esposito, G; Dhanapal, B; Dumy, P; Varma, V; Mutter, M; Bodenhausen, G

1997-01-01

The structure of the synthetic peptide CH3CO(Leu-Ser-Leu-Leu-Leu-Ser-Leu)3Lys-NH2 in trifluoroethanol/water 60/40 (volume ratio) was characterized by two-dimensional nmr spectroscopy. The peptide, closely related to the amphiphilic helix models designed by W. F. De-Grado and co-workers to mimic protein ion channels [(1988) Science, Vol. 240, p. 1177-1181], folds into a regular helix spanning residues 1-20. Evidence for a helix C-terminal capping conformation, involving the terminal lysine residue, was observed from Overhauser effects and checked for consistency by restrained molecular dynamics simulations. The side-chain amino group of Lys22 forms a hydrogen bond with the carbonyl of Leu18, and the distorted helical geometry of the terminal dipeptide allows the inclusion of a water bridge between the backbone NH of the Lys22 residue and the carbonyls of Leu19 and Ser20.

Thermodynamic Effects of Replacements of Pro Residues in Helix Interiors of Maltose-Binding Protein

OpenAIRE

Prajapati, RS; Lingaraju, GM; Bacchawat, Kiran; Surolia, Avadhesha; Varadarajan, Raghavan

2003-01-01

Introduction of Pro residues into helix interiors results in protein destabilization. It is currently unclear if the converse substitution (i.e., replacement of Pro residues that naturally occur in helix interiors would be stabilizing). Maltose-binding protein is a large 370-amino acid protein that contains 21 Pro residues. Of these, three nonconserved residues (P48, P133, and P159) occur at helix interiors. Each of the residues was replaced with Ala and Ser. Stabilities were characterized by...

Dual-purpose linker for alpha helix stabilization and imaging agent conjugation to glucagon-like peptide-1 receptor ligands.

Science.gov (United States)

Zhang, Liang; Navaratna, Tejas; Liao, Jianshan; Thurber, Greg M

2015-02-18

Peptides display many characteristics of efficient imaging agents such as rapid targeting, fast background clearance, and low non-specific cellular uptake. However, poor stability, low affinity, and loss of binding after labeling often preclude their use in vivo. Using glucagon-like peptide-1 receptor (GLP-1R) ligands exendin and GLP-1 as a model system, we designed a novel α-helix-stabilizing linker to simultaneously address these limitations. The stabilized and labeled peptides showed an increase in helicity, improved protease resistance, negligible loss or an improvement in binding affinity, and excellent in vivo targeting. The ease of incorporating azidohomoalanine in peptides and efficient reaction with the dialkyne linker enable this technique to potentially be used as a general method for labeling α helices. This strategy should be useful for imaging beta cells in diabetes research and in developing and testing other peptide targeting agents.

Eukaryotic beta-alanine synthases are functionally related but have a high degree of structural diversity

DEFF Research Database (Denmark)

Gojkovic, Zoran; Sandrini, Michael; Piskur, Jure

2001-01-01

no pyrimidine catabolic pathway, it enabled growth on N-carbamyl- beta -alanine as the sole nitrogen source. The D. discoideum and D. melanogaster PYD3 gene products are similar to mammalian beta -alanine synthases. In contrast, the S. kluyveri protein is quite different from these and more similar to bacterial......beta -Alanine synthase (EC 3.5.1.6), which catalyzes the final step of pyrimidine catabolism, has only been characterized in mammals. A Saccharomyces kluyveri pyd3 mutant that is unable to grow on N-carbamy-beta -alanine as the sole nitrogen source and exhibits diminished beta -alanine synthase...... N- carbamyl amidohydrolases. All three beta -alanine synthases are to some degree related to various aspartate transcarbamylases, which catalyze the second step of the de novo pyrimidine biosynthetic pathway. PYD3 expression in yeast seems to be inducible by dihydrouracil and N...

Electron Paramagnetic Resonance signal ratio of Alanine Pellets In Nuclear Power Plants

International Nuclear Information System (INIS)

Choi, Hoon; Sung, In Bok; Lee, Byung Il; Lim, Young Ki

2011-01-01

As a dosimeter for ESR dosimetry, alanine has many useful features including relatively long endurance time of radicals and almost no difference with the radiation dose rate and radiation quality. Alanine dosimeters have been accepted as transfer dosimeters for their good precision at high radiation range. For alanine/ESR spectra, it was reported that the peaks of the spectra are due to the three kinds of radicals induced by radiation. The ratio of the weak 'satellite line' and the central peak of the three man in alanine specta(in this study x /y ratio ) are changed on the LET value of radiation. In case of lithium formate monohydrate was recently identified as a promising dosimetric material. in addition, it was reported the the peak height in the signal spectra is not easily saturated when power is irradiated with relatively high LET radiation such as neutron rays. The difference in the peak height ratio was reported to be caused by increased local radical density following the radiation of high LET. The spectrum shape of some alanine dosimeter installed in the containment buildings of NPPs showed differences in comparison with dosimeters exposed only to gamma rays. There was apparent change of spectra, expressed as the 'x/y ratio'. As noted in other papers, high LET radiation such as neutron rays causes shape changes of the spectrum of alanine dosimeters. Thus, the unanticipated high dose level and low 'x/y ratio' of some alanine dosimeters from the containment building could be explained b exposure to mixed radiation with high LET. Generally, the locations evaluated in this study are regarded as being exposed only to gamma rays, because the positions are blocked from direct neutron rays, because the positions are blocked from direct neutron rays from reactor by thick cement barriers and heavy instruments.

One Peptide Reveals the Two Faces of α-Helix Unfolding-Folding Dynamics.

Science.gov (United States)

Jesus, Catarina S H; Cruz, Pedro F; Arnaut, Luis G; Brito, Rui M M; Serpa, Carlos

2018-04-12

The understanding of fast folding dynamics of single α-helices comes mostly from studies on rationally designed peptides displaying sequences with high helical propensity. The folding/unfolding dynamics and energetics of α-helix conformations in naturally occurring peptides remains largely unexplored. Here we report the study of a protein fragment analogue of the C-peptide from bovine pancreatic ribonuclease-A, RN80, a 13-amino acid residue peptide that adopts a highly populated helical conformation in aqueous solution. 1 H NMR and CD structural studies of RN80 showed that α-helix formation displays a pH-dependent bell-shaped curve, with a maximum near pH 5, and a large decrease in helical content in alkaline pH. The main forces stabilizing this short α-helix were identified as a salt bridge formed between Glu-2 and Arg-10 and the cation-π interaction involving Tyr-8 and His-12. Thus, deprotonation of Glu-2 or protonation of His-12 are essential for the RN80 α-helix stability. In the present study, RN80 folding and unfolding were triggered by laser-induced pH jumps and detected by time-resolved photoacoustic calorimetry (PAC). The photoacid proton release, amino acid residue protonation, and unfolding/folding events occur at different time scales and were clearly distinguished using time-resolved PAC. The partial unfolding of the RN80 α-helix, due to protonation of Glu-2 and consequent breaking of the stabilizing salt bridge between Glu-2 and Arg-10, is characterized by a concentration-independent volume expansion in the sub-microsecond time range (0.8 mL mol -1 , 369 ns). This small volume expansion reports the cost of peptide backbone rehydration upon disruption of a solvent-exposed salt bridge, as well as backbone intrinsic expansion. On the other hand, RN80 α-helix folding triggered by His-12 protonation and subsequent formation of a cation-π interaction leads to a microsecond volume contraction (-6.0 mL mol -1 , ∼1.7 μs). The essential role of two

[A systematic review of biosynthesis of poly (3-hydroxypropionate)].

Science.gov (United States)

Chang, Le; Zhan, Yuanlong; Liu, Changli

2018-04-25

Poly (3-hydroxypropionate) (P3HP), a new member of thermoplastic of family polyhydroxyalkanoates (PHAs), has excellent characteristics of biodegradability and biocompatibility. By now no reports can be found about wild-type bacteria that naturally synthesize P3HP, so the main way to produce P3HP is chemical and biological methods. Chemical method by adding high cost 3-HP monomers or their structural analogs as precursors, has the drawbacks of toxicity, low effectiveness and high cost. Biological method using engineered strain may utilize inexpensive and renewable carbon source to produce P3HP and has gradually become more and more popular. We systematically review here the biosynthesis of P3HP research progress. The advantages and disadvantages of biosynthesis pathways of glycerol pathway, malonyl-CoA pathway and β-alanine pathway were analyzed.

Conformational Diffusion and Helix Formation Kinetics

International Nuclear Information System (INIS)

Hummer, Gerhard; Garcia, Angel E.; Garde, Shekhar

2000-01-01

The time, temperature, and sequence dependences of helix formation kinetics of fully atomistic peptide models in explicit solvent are described quantitatively by a diffusive search within the coil state with barrierless transitions into the helical state. Conformational diffusion leads to nonexponential kinetics and jump-width dependences in temperature jump experiments. (c) 2000 The American Physical Society

Conformational Diffusion and Helix Formation Kinetics

Energy Technology Data Exchange (ETDEWEB)

Hummer, Gerhard [Laboratory of Chemical Physics, Building 5, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892-0520 (United States); Garcia, Angel E. [Theoretical Biology and Biophysics Group T-10, MS K710, Los Alamos National Laboratory, Los Alamos, New Mexico 87545 (United States); Garde, Shekhar [Department of Chemical Engineering, Rensselaer Polytechnic Institute, 110 8th Street, Troy, New York 12180 (United States)

2000-09-18

The time, temperature, and sequence dependences of helix formation kinetics of fully atomistic peptide models in explicit solvent are described quantitatively by a diffusive search within the coil state with barrierless transitions into the helical state. Conformational diffusion leads to nonexponential kinetics and jump-width dependences in temperature jump experiments. (c) 2000 The American Physical Society.

Disruption of the LOV-Jalpha helix interaction activates phototropin kinase activity.

Science.gov (United States)

Harper, Shannon M; Christie, John M; Gardner, Kevin H

2004-12-28

Light plays a crucial role in activating phototropins, a class of plant photoreceptors that are sensitive to blue and UV-A wavelengths. Previous studies indicated that phototropin uses a bound flavin mononucleotide (FMN) within its light-oxygen-voltage (LOV) domain to generate a protein-flavin covalent bond under illumination. In the C-terminal LOV2 domain of Avena sativa phototropin 1, formation of this bond triggers a conformational change that results in unfolding of a helix external to this domain called Jalpha [Harper, S. M., et al. (2003) Science 301, 1541-1545]. Though the structural effects of illumination were characterized, it was unknown how these changes are coupled to kinase activation. To examine this, we made a series of point mutations along the Jalpha helix to disrupt its interaction with the LOV domain in a manner analogous to light activation. Using NMR spectroscopy and limited proteolysis, we demonstrate that several of these mutations displace the Jalpha helix from the LOV domain independently of illumination. When placed into the full-length phototropin protein, these point mutations display constitutive kinase activation, without illumination of the sample. These results indicate that unfolding of the Jalpha helix is the critical event in regulation of kinase signaling for the phototropin proteins.

An efficient protocol for the complete incorporation of methyl-protonated alanine in perdeuterated protein

International Nuclear Information System (INIS)

Ayala, Isabel; Sounier, Remy; Use, Nathalie; Gans, Pierre; Boisbouvier, Jerome

2009-01-01

A strategy for the introduction of ( 1 H, 13 C-methyl)-alanine into perdeuterated proteins is described. Specific protonation of alanine methyl groups to a level of 95% can be achieved by overexpressing proteins in M9/D 2 O based bacterial growth medium supplemented with 800 mg/l of 2-[ 2 H], 3-[ 13 C] l-alanine. However, though simple, this approach results in undesired, non-specific background labeling due to isotope scrambling via different amino acid metabolic pathways. Following a careful analysis of known metabolic pathways we found that co-addition of perdeuterated forms of α-ketoisovalerate-d 7 , succinate-d 4 and l-isoleucine-d 10 with labeled l-alanine, reduces undesired background labeling to <1%. When combined with recently developed methyl TROSY experiments, this methyl-specific labeling protocol permits the acquisition of excellent quality correlation spectra of alanine methyl groups in high molecular weight proteins. Our cost effective strategy offers a significant enhancement in the level of incorporation of methyl-labeled alanine in overexpressed proteins over previously reported methods

Polymerization of alanine in the presence of a non-swelling montmorillonite

Science.gov (United States)

Paecht-Horowitz, M.; Lahav, N.

1977-01-01

Alanine, starting from alanine-adenylate, has been polymerized in the presence of non-swelling Al-montmorillonite. The yield of polymerization is much lower than that obtained in the presence of swelling Na-montmorillonite. The possibility that the changing interlayer spacing in Na-montmorillonite might be responsible for its catalytic properties, is discussed.

A thin alanine-polyethylene film dosimetry system with diffuse reflection spectrophotometric evaluation

International Nuclear Information System (INIS)

Zagorski, Z.P.; Rafalski, A.

1995-01-01

Characteristics of a new alanine dosimeter in the shape of a thin film, with the measurement of optical absorption of the CH 3 CHCOO - radical is described. That type of dosimeter, ALA/DRS (for diffuse reflection spectrophotometry) is compared, to an alanine dosimeter with EPR evaluation (ALA/EPR for short). In many respects the simple ALA/DRS version, as the alanine-polyethylene composite is superior. The paper shows the importance of the new experimental approach to free radical research in solid state radiation chemistry. (author). 7 refs., 3 figs

Degradation of pyrimidines in Saccharomyces kluyveri: transamination of beta-alanine

DEFF Research Database (Denmark)

Schnackerz, K D; Andersen, G; Dobritzsch, D

2008-01-01

Beta-alanine is an intermediate in the reductive degradation of uracil. Recently we have identified and characterized the Saccharomyces kluyveri PYD4 gene and the corresponding enzyme beta -alanine aminotransferase ((Sk)Pyd4p), highly homologous to eukaryotic gamma-aminobutyrate aminotransferase ...

13-Helix folding of a β/γ-peptide manifold designed from a "minimal-constraint" blueprint.

Science.gov (United States)

Grison, Claire M; Robin, Sylvie; Aitken, David J

2016-06-14

A bottom-up design rationale was adopted to devise β/γ-peptide foldamer manifolds which would adopt preferred 13-helix conformations, relying on minimal steric imposition brought by the constituent amino acid residues. In this way, a well-defined 13-helix conformer was revealed for short oligomers of trans-2-aminocyclobutanecarboxylic acid and γ(4)-amino acids in alternation, which gave good topological superposition upon an α-helix motif.

Relative response of the alanine dosimeter to medium energy x-rays.

Science.gov (United States)

Anton, M; Büermann, L

2015-08-07

The response of the alanine dosimeter to kilovoltage x-rays with respect to the dose to water was measured, relative to the response to Co-60 radiation.Two series of x-ray qualities were investigated, one ranging from 30 kV to 100 kV tube voltage (TW series), the other one ranging from 70 kV to 280 kV (TH series). Due to the use of the water calorimeter as a primary standard, the uncertainty of the delivered dose is significantly lower than for other published data. The alanine response was measured as described in a previous publication (Anton et al 2013 Phys. Med. Biol. 58 3259-82). The uncertainty component due to the alanine measurement and analysis is ⩽0.4%, the major part of the combined uncertainty of the relative response originates from the uncertainty of the delivered dose. The relative uncertainties of the relative response vary from ⩽2% for the TW series to ⩽1.1% for the TH series.Different from the behaviour of the alanine dosimeter for megavoltage x-rays or electrons, the relative response drops significantly from unity for Co-60 radiation to less than 64% for the TW quality with a tube voltage of 30 kV. In order to reproduce this behaviour through Monte Carlo simulations, not only the ratio of the absorbed dose to alanine to the absorbed dose to water has to be known, but also the intrinsic efficiency, i.e. the dependence of the number of free radicals generated per unit of absorbed dose on the photon energy. This quantity is not yet accessible for the TW series.For a possible use of the alanine dosimeter for kilovoltage x-rays, for example in electronic brachytherapy, users should rely on the measured data for the relative response which have become available with this publication.

End-to-end tests using alanine dosimetry in scanned proton beams

Science.gov (United States)

Carlino, A.; Gouldstone, C.; Kragl, G.; Traneus, E.; Marrale, M.; Vatnitsky, S.; Stock, M.; Palmans, H.

2018-03-01

This paper describes end-to-end test procedures as the last fundamental step of medical commissioning before starting clinical operation of the MedAustron synchrotron-based pencil beam scanning (PBS) therapy facility with protons. One in-house homogeneous phantom and two anthropomorphic heterogeneous (head and pelvis) phantoms were used for end-to-end tests at MedAustron. The phantoms were equipped with alanine detectors, radiochromic films and ionization chambers. The correction for the ‘quenching’ effect of alanine pellets was implemented in the Monte Carlo platform of the evaluation version of RayStation TPS. During the end-to-end tests, the phantoms were transferred through the workflow like real patients to simulate the entire clinical workflow: immobilization, imaging, treatment planning and dose delivery. Different clinical scenarios of increasing complexity were simulated: delivery of a single beam, two oblique beams without and with range shifter. In addition to the dose comparison in the plastic phantoms the dose obtained from alanine pellet readings was compared with the dose determined with the Farmer ionization chamber in water. A consistent systematic deviation of about 2% was found between alanine dosimetry and the ionization chamber dosimetry in water and plastic materials. Acceptable agreement of planned and delivered doses was observed together with consistent and reproducible results of the end-to-end testing performed with different dosimetric techniques (alanine detectors, ionization chambers and EBT3 radiochromic films). The results confirmed the adequate implementation and integration of the new PBS technology at MedAustron. This work demonstrates that alanine pellets are suitable detectors for end-to-end tests in proton beam therapy and the developed procedures with customized anthropomorphic phantoms can be used to support implementation of PBS technology in clinical practice.

Reovirus FAST Proteins Drive Pore Formation and Syncytiogenesis Using a Novel Helix-Loop-Helix Fusion-Inducing Lipid Packing Sensor.

Directory of Open Access Journals (Sweden)

Jolene Read

2015-06-01

Full Text Available Pore formation is the most energy-demanding step during virus-induced membrane fusion, where high curvature of the fusion pore rim increases the spacing between lipid headgroups, exposing the hydrophobic interior of the membrane to water. How protein fusogens breach this thermodynamic barrier to pore formation is unclear. We identified a novel fusion-inducing lipid packing sensor (FLiPS in the cytosolic endodomain of the baboon reovirus p15 fusion-associated small transmembrane (FAST protein that is essential for pore formation during cell-cell fusion and syncytiogenesis. NMR spectroscopy and mutational studies indicate the dependence of this FLiPS on a hydrophobic helix-loop-helix structure. Biochemical and biophysical assays reveal the p15 FLiPS preferentially partitions into membranes with high positive curvature, and this partitioning is impeded by bis-ANS, a small molecule that inserts into hydrophobic defects in membranes. Most notably, the p15 FLiPS can be functionally replaced by heterologous amphipathic lipid packing sensors (ALPS but not by other membrane-interactive amphipathic helices. Furthermore, a previously unrecognized amphipathic helix in the cytosolic domain of the reptilian reovirus p14 FAST protein can functionally replace the p15 FLiPS, and is itself replaceable by a heterologous ALPS motif. Anchored near the cytoplasmic leaflet by the FAST protein transmembrane domain, the FLiPS is perfectly positioned to insert into hydrophobic defects that begin to appear in the highly curved rim of nascent fusion pores, thereby lowering the energy barrier to stable pore formation.

Photoluminescence Enhancement of Poly(3-methylthiophene Nanowires upon Length Variable DNA Hybridization

Directory of Open Access Journals (Sweden)

Jingyuan Huang

2018-01-01

Full Text Available The use of low-dimensional inorganic or organic nanomaterials has advantages for DNA and protein recognition due to their sensitivity, accuracy, and physical size matching. In this research, poly(3-methylthiophene (P3MT nanowires (NWs are electrochemically prepared with dopant followed by functionalization with probe DNA (pDNA sequence through electrostatic interaction. Various lengths of pDNA sequences (10-, 20- and 30-mer are conjugated to the P3MT NWs respectively followed with hybridization with their complementary target DNA (tDNA sequences. The nanoscale photoluminescence (PL properties of the P3MT NWs are studied throughout the whole process at solid state. In addition, the correlation between the PL enhancement and the double helix DNA with various lengths is demonstrated.

Geometry of the toroidal N-helix: optimal-packing and zero-twist

DEFF Research Database (Denmark)

Olsen, Kasper; Bohr, Jakob

2012-01-01

Two important geometrical properties of N-helix structures are influenced by bending. One is maximizing the volume fraction, which is called optimal-packing, and the other is having a vanishing strain-twist coupling, which is called zero-twist. Zero-twist helices rotate neither in one nor...... helix. General N-helices are discussed, as well as zero-twist helices for N > 1. The derived geometrical restrictions are gradually modified by changing the aspect ratio of the torus....

Lack of Effect of Sodium Benzoate at Reported Clinical Therapeutic Concentration on d-Alanine Metabolism in Dogs.

Science.gov (United States)

Popiolek, Michael; Tierney, Brendan; Steyn, Stefanus J; DeVivo, Michael

2018-06-19

Cognitive decline and psychosis have been hypothesized to be mediated by N-methyl-d-aspartate receptor (NMDAR) hypofunction. Consistent with this hypothesis, chronic treatment with d-alanine, a coagonist at the glycine site of the NMDAR, leads to an improvement of positive and cognitive symptoms in schizophrenic patients. d-alanine is oxidized by d-amino acid oxidase (DAAO); thus, an inhibitor of DAAO would be expected to enhance d-alanine levels and likewise lead to desirable clinical outcomes. Sodium benzoate, on the basis of d-amino acid inhibition, was observed to display beneficial clinical effects in schizophrenic and Alzheimer's patients. However, in the clinical pilot studies using sodium benzoate, d-amino acids were not quantified to verify that sodium benzoate's efficacy was mediated through DAAO inhibition. In this study, d-alanine content was monitored in cerebral spinal fluid (CSF) of dogs treated with daily injections of d-alanine (30 mg/kg) alone and in combination with sodium benzoate (30 mg/kg) for seven consecutive days. We reasoned that the cerebral spinal fluid d-alanine quantity is reflective of the brain d-alanine levels and it would increase as a consequence of DAAO inhibition with sodium benzoate. We found that d-alanine treatment lead to maximal concentration of 7.51 μM CSF d-alanine level; however, coadministration of sodium benzoate and d-alanine did not change CSF d-alanine level beyond that of d-alanine treatment alone. As a consequence, we conclude that clinical efficacy associated with chronic administration of sodium benzoate in schizophrenic and Alzheimer's patients is likely not mediated through inhibition of DAAO.

Carnosine and anserine homeostasis in skeletal muscle and heart is controlled by β‐alanine transamination

Science.gov (United States)

Blancquaert, Laura; Baba, Shahid P.; Kwiatkowski, Sebastian; Stautemas, Jan; Stegen, Sanne; Barbaresi, Silvia; Chung, Weiliang; Boakye, Adjoa A.; Hoetker, J. David; Bhatnagar, Aruni; Delanghe, Joris; Vanheel, Bert; Veiga‐da‐Cunha, Maria; Derave, Wim

2016-01-01

Key points Using recombinant DNA technology, the present study provides the first strong and direct evidence indicating that β‐alanine is an efficient substrate for the mammalian transaminating enzymes 4‐aminobutyrate‐2‐oxoglutarate transaminase and alanine‐glyoxylate transaminase.The concentration of carnosine and anserine in murine skeletal and heart muscle depends on circulating availability of β‐alanine, which is in turn controlled by degradation of β‐alanine in liver and kidney.Chronic oral β‐alanine supplementation is a popular ergogenic strategy in sports because it can increase the intracellular carnosine concentration and subsequently improve the performance of high‐intensity exercises. The present study can partly explain why the β‐alanine supplementation protocol is so inefficient, by demonstrating that exogenous β‐alanine can be effectively routed toward oxidation. Abstract The metabolic fate of orally ingested β‐alanine is largely unknown. Chronic β‐alanine supplementation is becoming increasingly popular for improving high‐intensity exercise performance because it is the rate‐limiting precursor of the dipeptide carnosine (β‐alanyl‐l‐histidine) in muscle. However, only a small fraction (3–6%) of the ingested β‐alanine is used for carnosine synthesis. Thus, the present study aimed to investigate the putative contribution of two β‐alanine transamination enzymes, namely 4‐aminobutyrate‐2‐oxoglutarate transaminase (GABA‐T) and alanine‐glyoxylate transaminase (AGXT2), to the homeostasis of carnosine and its methylated analogue anserine. We found that, when transfected into HEK293T cells, recombinant mouse and human GABA‐T and AGXT2 are able to transaminate β‐alanine efficiently. The reaction catalysed by GABA‐T is inhibited by vigabatrin, whereas both GABA‐T and AGXT2 activity is inhibited by aminooxyacetic acid (AOA). Both GABA‐T and AGXT2 are highly expressed in the mouse liver and

[Alanine solution as enzyme reaction buffer used in A to O blood group conversion].

Science.gov (United States)

Li, Su-Bo; Zhang, Xue; Zhang, Yin-Ze; Tan, Ying-Xia; Bao, Guo-Qiang; Wang, Ying-Li; Ji, Shou-Ping; Gong, Feng; Gao, Hong-Wei

2014-06-01

The aim of this study was to investigate the effect of alanine solution as α-N-acetylgalactosaminidase enzyme reaction buffer on the enzymatic activity of A antigen. The binding ability of α-N-acetylgalactosaminidase with RBC in different reaction buffer such as alanine solution, glycine solution, normal saline (0.9% NaCl), PBS, PCS was detected by Western blot. The results showed that the efficiency of A to O conversion in alanine solution was similar to that in glycine solution, and Western blot confirmed that most of enzymes blinded with RBC in glycine or alanine solution, but few enzymes blinded with RBC in PBS, PCS or normal saline. The evidences indicated that binding of enzyme with RBC was a key element for A to O blood group conversion, while the binding ability of α-N-acetylgalactosaminidase with RBC in alanine or glycine solution was similar. It is concluded that alanine solution can be used as enzyme reaction buffer in A to O blood group conversion. In this buffer, the α-N-acetylgalactosaminidase is closely blinded with RBC and α-N-acetylgalactosaminidase plays efficient enzymatic activity of A antigen.

Physiological hypercortisolemia increases proteolysis, glutamine, and alanine production

International Nuclear Information System (INIS)

Darmaun, D.; Matthews, D.E.; Bier, D.M.

1988-01-01

Physiological elevations of plasma cortisol levels, as are encountered in stress and severe trauma, were produced in six normal subjects by infusing them with hydrocortisone for 64 h. Amino acid kinetics were measured in the postabsorptive state using three 4-h infusions of L-[1- 13 C]leucine, L-phenyl[ 2 H 5 ]phenylalanine, L-[2- 15 N]glutamine, and L-[1- 13 C]alanine tracers (1) before, (2) at 12 h, and (3) at 60 h of cortisol infusion. Before and throughout the study, the subjects ate a normal diet of adequate protein and energy intake. The cortisol infusion raised plasma cortisol levels significantly from 10 ± 1 to 32 ± 4 μg/dl, leucine flux from 83 ± 3 to 97 ± 3 μmol·kg -1 ·h -1 , and phenylalanine flux from 34 ± 1 to 39 ± 1 (SE) μmol·kg -1 ·h -1 after 12 h of cortisol infusion. These increases were maintained until the cortisol infusion was terminated. These nearly identical 15% increases in two different essential amino acid appearance rates are reflective of increased whole body protein breakdown. Glutamine flux rose by 12 h of cortisol infusion and remained elevated at the same level at 64 h. The increase in flux was primarily due to a 55% increase in glutamine de novo synthesis. Alanine flux increased with acute hypercortisolemia and increased further at 60 h of cortisol infusion, a result primarily of increased alanine de novo synthesis. Insulin, alanine, and lactate plasma levels responded similarly with significant rises between the acute and chronic periods of cortisol infusion. Thus hypercortisolemia increases both protein breakdown and the turnover of important nonessential amino acids for periods of up to 64 h

Role of α-globin H helix in the building of tetrameric human hemoglobin: interaction with α-hemoglobin stabilizing protein (AHSP) and heme molecule.

Science.gov (United States)

Domingues-Hamdi, Elisa; Vasseur, Corinne; Fournier, Jean-Baptiste; Marden, Michael C; Wajcman, Henri; Baudin-Creuza, Véronique

2014-01-01

Alpha-Hemoglobin Stabilizing Protein (AHSP) binds to α-hemoglobin (α-Hb) or α-globin and maintains it in a soluble state until its association with the β-Hb chain partner to form Hb tetramers. AHSP specifically recognizes the G and H helices of α-Hb. To investigate the degree of interaction of the various regions of the α-globin H helix with AHSP, this interface was studied by stepwise elimination of regions of the α-globin H helix: five truncated α-Hbs α-Hb1-138, α-Hb1-134, α-Hb1-126, α-Hb1-123, α-Hb1-117 were co-expressed with AHSP as two glutathione-S-transferase (GST) fusion proteins. SDS-PAGE and Western Blot analysis revealed that the level of expression of each truncated α-Hb was similar to that of the wild type α-Hb except the shortest protein α-Hb1-117 which displayed a decreased expression. While truncated GST-α-Hb1-138 and GST-α-Hb1-134 were normally soluble; the shorter globins GST-α-Hb1-126 and GST-α-Hb1-117 were obtained in very low quantities, and the truncated GST-α-Hb1-123 provided the least material. Absorbance and fluorescence studies of complexes showed that the truncated α-Hb1-134 and shorter forms led to modified absorption spectra together with an increased fluorescence emission. This attests that shortening the H helix leads to a lower affinity of the α-globin for the heme. Upon addition of β-Hb, the increase in fluorescence indicates the replacement of AHSP by β-Hb. The CO binding kinetics of different truncated AHSPWT/α-Hb complexes showed that these Hbs were not functionally normal in terms of the allosteric transition. The N-terminal part of the H helix is primordial for interaction with AHSP and C-terminal part for interaction with heme, both features being required for stability of α-globin chain.

NMR solution structure of the mitochondrial F1beta presequence from Nicotiana plumbaginifolia.

Science.gov (United States)

Moberg, Per; Nilsson, Stefan; Ståhl, Annelie; Eriksson, Anna-Carin; Glaser, Elzbieta; Mäler, Lena

2004-03-05

We have isolated, characterized and determined the three-dimensional NMR solution structure of the presequence of ATPsynthase F1beta subunit from Nicotiana plumbaginifolia. A general method for purification of presequences is presented. The method is based on overexpression of a mutant precursor containing a methionine residue introduced at the processing site, followed by CNBr-cleavage and purification of the presequence on a cation-exchange column. The F1beta presequence, 53 amino acid residues long, retained its native properties as evidenced by inhibition of in vitro mitochondrial import and processing at micromolar concentrations. CD spectroscopy revealed that the F1beta presequence formed an alpha-helical structure in membrane mimetic environments such as SDS and DPC micelles (approximately 50% alpha-helix), and in acidic phospholipid bicelles (approximately 60% alpha-helix). The NMR solution structure of the F1beta presequence in SDS micelles was determined on the basis of 518 distance and 21 torsion angle constraints. The structure was found to contain two helices, an N-terminal amphipathic alpha-helix (residues 4-15) and a C-terminal alpha-helix (residues 43-53), separated by a largely unstructured 27 residue long internal domain. The N-terminal amphipathic alpha-helix forms the putative Tom20 receptor binding site, whereas the C-terminal alpha-helix is located upstream of the mitochondrial processing peptidase cleavage site.

Solid State Structure of Poly(9,9-dinonylfluorene)

DEFF Research Database (Denmark)

Torkkeli, Mika; Galbrecht, Frank; Scherf, Ullrich

2015-01-01

We report on X-ray diffraction and grazing-incidence X-ray diffraction data of poly(9,9-dinonylfluorene) (PF9) in bulk, thin films and in the 1% methylcyclohexane gel. We denote the main crystalline phase as alpha phase and propose that the unit cell is monoclinic (a = 29.31 angstrom, b = 23.......1 angstrom, and c = 16.7 angstrom). Structural analogues to other 9,9-di-n-alkyl-substituted polyfluorenes are discussed in terms of unit cell parameters and backbone geometry....

Spectrophotometric readout for alanine dosimeter to be used for food irradiation applications

International Nuclear Information System (INIS)

Ebraheem, S.; Beshir, W.B.; Eid, S.

2002-01-01

The electron spin resonance (EPR) readout of radical concentration produced upon irradiation of L-alanine is well known as a transfer dosimetry system for high dose level. The highly cost of EPR/alanine dosimetry system is a serious handicap for large scale routine application to be used in irradiation facilities. In this study, the reaction between L-alanine and 1,4-phenyl diammonium dichloride (PAC) solution produces a complex has a purple color. This complex has a variable absorbance with applied doses in the range from 1-20 kGy. Spectrophotometric evaluation at 368 and 505 nm for the absorbance intensity of this color as well as fluorimetric emission wavelength 435 nm were investigated, as a function of dose. The used method is uncomplicated and very easy for routine application. Evaluation of the dye concentration and the suitable amount of alanine has been studied. Stability of the product complex, for long period of time after the reaction had taken place, was also investigated

14N nuclear quadrupole interaction in Cu(II) doped L-alanine

International Nuclear Information System (INIS)

Murgich, J.; Calvo, R.; Oseroff, S.B.; Instituto Venezolano de Investigaciones Cientificas, Caracas. Dept. de Quimica)

1980-01-01

The 14 N nuclear quadrupole interaction tensor Psub(N) measured by ENDOR in Cu(II) doped L-alanine is analyzed in terms of the Townes and Daily theory assuming a tetra-hedrally bonded N atom. The results of this analysis are compared with those for the 14 N in pure L-alanine and it is found that the principal directions of the Psub(N) tensor are drastically changed upon metal complexation as a consequence of the higher electron affinity of Cu(II) with respect to C and H. Comparison of the corresponding bond populations in pure and Cu(II) doped L-alanine indicates that the Cu draws 0.11 more electron from the N than the substituted H atom. (orig.)

Revised mechanism of d-alanine incorporation into cell wall polymers in Gram-positive bacteria

Science.gov (United States)

Reichmann, Nathalie T.; Cassona, Carolina Picarra

2013-01-01

Teichoic acids (TAs) are important for growth, biofilm formation, adhesion and virulence of Gram-positive bacterial pathogens. The chemical structures of the TAs vary between bacteria, though they typically consist of zwitterionic polymers that are anchored to either the peptidoglycan layer as in the case of wall teichoic acid (WTA) or the cell membrane and named lipoteichoic acid (LTA). The polymers are modified with d-alanines and a lack of this decoration leads to increased susceptibility to cationic antimicrobial peptides. Four proteins, DltA–D, are essential for the incorporation of d-alanines into cell wall polymers and it has been established that DltA transfers d-alanines in the cytoplasm of the cell onto the carrier protein DltC. However, two conflicting models have been proposed for the remainder of the mechanism. Using a cellular protein localization and membrane topology analysis, we show here that DltC does not traverse the membrane and that DltD is anchored to the outside of the cell. These data are in agreement with the originally proposed model for d-alanine incorporation through a process that has been proposed to proceed via a d-alanine undecaprenyl phosphate membrane intermediate. Furthermore, we found that WTA isolated from a Staphylococcus aureus strain lacking LTA contains only a small amount of d-alanine, indicating that LTA has a role, either direct or indirect, in the efficient d-alanine incorporation into WTA in living cells. PMID:23858088

Rapid mapping of protein functional epitopes by combinatorial alanine scanning

OpenAIRE

Weiss, GA; Watanabe, CK; Zhong, A; Goddard, A; Sidhu, SS

2000-01-01

A combinatorial alanine-scanning strategy was used to determine simultaneously the functional contributions of 19 side chains buried at the interface between human growth hormone and the extracellular domain of its receptor. A phage-displayed protein library was constructed in which the 19 side chains were preferentially allowed to vary only as the wild type or alanine. The library pool was subjected to binding selections to isolate functional clones, and DNA sequencing was used to determine ...

Mechanism of mercurial inhibition of sodium-coupled alanine uptake in liver plasma membrane vesicles from Raja erinacea

International Nuclear Information System (INIS)

Sellinger, M.; Ballatori, N.; Boyer, J.L.

1991-01-01

In mammalian hepatocytes the L-alanine carrier contains a sulfhydryl group that is essential for its activity and is inhibited by mercurials. In hepatocytes of the evolutionarily primitive little skate (Raja erinacea), HgCl2 inhibits Na(+)-dependent alanine uptake and Na+/K(+)-ATPase and increase K+ permeability. To distinguish between direct effects of HgCl2 on the Na(+)-alanine cotransporter and indirect effects on membrane permeability, [3H]alanine transport was studied in plasma membrane vesicles. [3H]Alanine uptake was stimulated by an out-to-in Na+ but not K+ gradient and was saturable confirming the presence of Na(+)-alanine cotransport in liver plasma membranes from this species. Preincubation of the vesicles with HgCl2 for 5 min reduced initial rates of Na(+)-dependent but not Na(+)-independent alanine uptake in a dose-dependent manner (10-200 microM). In the presence of equal concentrations of NaCl or KCl inside and outside of the vesicles, 75 microM HgCl2 directly inhibited sodium-dependent alanine-[3H]alanine exchange, demonstrating that HgCl2 directly affected the alanine cotransporter. Inhibition of Na(+)-dependent alanine uptake by 30 microM HgCl2 was reversed by dithiothreitol (1 mM). HgCl2 (10-30 microM) also increased initial rates of 22Na uptake (at 5 sec), whereas 22Na uptake rates were decreased at HgCl2 concentrations greater than 50 microM. Higher concentrations of HgCl2 (100-200 microM) produced nonspecific effects on vesicle integrity. These studies indicate that HgCl2 inhibits Na(+)-dependent alanine uptake in skate hepatocytes by three different concentration-dependent mechanisms: direct interaction with the transporters, dissipation of the driving force (Na+ gradient), and loss of membrane integrity

The application of Alanine/ESR dosimetry in nuclear power plants

Energy Technology Data Exchange (ETDEWEB)

Choi, Hoon; Lee, Byung Il [Radiation Health Research Institute, Seoul (Korea, Republic of)

2014-05-15

Alanine/Electron spin resonance(ESR) has been proven very effective tool which dosimetric characteristics is better suitable than generally used personnel dosimeter for long term dose estimation. L-α-alanine has unusual stability of radiation induced radicals. The fading is known as about 1% a year by IAEA technical documentary. It also has linear signal response on gamma rays over the wide range of dose quantity. Alanine is a kind of unnecessary amino acid in 20 amino acids. Estimation of the accumulated gamma dose is important to predict the life expectancy of cables. However, exact estimation of gamma dose at containment building of NPP is very difficult, because the variability of estimation value is apparently depending on the each installation position in containment building. Especially, some installation positions near reactor change extremely. So, the data from ESR measurement should be checked to the details on referring installation map and pictures.

Progesterone modulation of transmembrane helix-helix interactions between the α-subunit of Na/K-ATPase and phospholipid N-methyltransferase in the oocyte plasma membrane

Directory of Open Access Journals (Sweden)

Askari Amir

2010-05-01

Full Text Available Abstract Background Progesterone binding to the surface of the amphibian oocyte initiates the meiotic divisions. Our previous studies with Rana pipiens oocytes indicate that progesterone binds to a plasma membrane site within the external loop between the M1 and M2 helices of the α-subunit of Na/K-ATPase, triggering a cascade of lipid second messengers and the release of the block at meiotic prophase. We have characterized this site, using a low affinity ouabain binding isoform of the α1-subunit. Results Preparations of isolated plasma membranes from Rana oocytes demonstrate that physiological levels of progesterone (or the non-metabolizable progestin R5020 successively activate phosphatidylethanolamine-N-methyltransferase (PE-NMT and sphingomyelin synthase within seconds. Inhibition of PE-NMT blocks the progesterone induction of meiosis in intact oocytes, whereas its initial product, phosphatidylmonomethylethanolamine (PME, can itself initiate meiosis in the presence of the inhibitor. Published X-ray crystallographic data on Na/K-ATPase, computer-generated 3D projections, heptad repeat analysis and hydrophobic cluster analysis of the transmembrane helices predict that hydrophobic residues L, V, V, I, F and Y of helix M2 of the α1-subunit interact with F, L, G, L, L and F, respectively, of helix M3 of PE-NMT. Conclusion We propose that progesterone binding to the first external loop of the α1-subunit facilitates specific helix-helix interactions between integral membrane proteins to up-regulate PE-NMT, and, that successive interactions between two or more integral plasma membrane proteins induce the signaling cascades which result in completion of the meiotic divisions.

Optical signal response pf the alanine gel solution for photons and electrons clinical beams

International Nuclear Information System (INIS)

Silva, Cleber Feijo; Campos, Leticia Lucente

2009-01-01

Alanine gel dosimeter is a new gel material developed at IPEN that presents significant improvement on previous alanine systems developed by Costa (1994). The measure technique is based on the transformation of ferrous ions (Fe 2+ ) in ferric ions (Fe 3+ ) after irradiation. The DL-Alanine (C 3 H 7 NO 2 ) is an aminoacid tissue equivalent that improves the production of ferric ions in the solution. This work aims to study the comparison of optical signal response of the alanine gel solution for photons and electrons clinical beams. It was observed that the calibration factor can be considered independent of quality of the radiation for photons and electrons clinical beams. Therefore, it can be used the same calibration factor for evaluating the absorbed dose in photons and electrons fields in the energy of 6 MeV. Alanine Gel Dosimeter presents good performance and can be useful as alternative dosimeter in the radiotherapy area using MRI technique for 3D dose distribution evaluation. (author)

ald of Mycobacterium tuberculosis Encodes both the Alanine Dehydrogenase and the Putative Glycine Dehydrogenase

Science.gov (United States)

Giffin, Michelle M.; Modesti, Lucia; Raab, Ronald W.; Wayne, Lawrence G.

2012-01-01

The putative glycine dehydrogenase of Mycobacterium tuberculosis catalyzes the reductive amination of glyoxylate to glycine but not the reverse reaction. The enzyme was purified and identified as the previously characterized alanine dehydrogenase. The Ald enzyme was expressed in Escherichia coli and had both pyruvate and glyoxylate aminating activities. The gene, ald, was inactivated in M. tuberculosis, which resulted in the loss of all activities. Both enzyme activities were found associated with the cell and were not detected in the extracellular filtrate. By using an anti-Ald antibody, the protein was localized to the cell membrane, with a smaller fraction in the cytosol. None was detected in the extracellular medium. The ald knockout strain grew without alanine or glycine and was able to utilize glycine but not alanine as a nitrogen source. Transcription of ald was induced when alanine was the sole nitrogen source, and higher levels of Ald enzyme were measured. Ald is proposed to have several functions, including ammonium incorporation and alanine breakdown. PMID:22210765

L-alanine-induced germination in Bacillus licheniformis -the impact of native gerA sequences.

Science.gov (United States)

Madslien, Elisabeth H; Granum, Per Einar; Blatny, Janet M; Lindbäck, Toril

2014-04-22

L-alanine, acting through the GerA receptor, was recently found to be an efficient germinant in Bacillus licheniformis ATCC14580/DSM13. In this study, we show that several of 46 examined B. licheniformis strains germinate remarkably slower than the type strain when exposed to L-alanine. These strains are not necessarily closely related, as determined by MLST (multi-locus sequence typing). Three of the slow-germinating strains were further examined in order to see whether nucleotide substitutions in the gerA sequences were responsible for the slow L-alanine germination. This was performed by complementing the transformable type strain derivate MW3ΔgerAA with gerA variants from the three slow-germinating strains; NVH1032, NVH1112 and NVH800. A wide selection of B. licheniformis strains was evaluated for L-alanine-induced germination efficiency. Our results show that gerA substitutions could only partially explain why spores of some B. licheniformis strains responded slower than others in the presence of L-alanine.

Radical formation of irradiated α-alanine and N-acetyl alanine with heavy ion beams. Effects of the irradiation temperature

International Nuclear Information System (INIS)

Minegishi, Atsuko; Nagasaki, Jun; Mori, Wasuke; Amano, Chikara; Takagi, Shinji; Murakami, Takeshi; Kanai, Tatsuaki; Furusawa, Yoshiya; Iwata, Yoshiyuki

2003-01-01

The characteristics of irradiation with C290 MeV/u ion beams were investigated using X-band electron spin resonance (ESR) spectroscopy for a polycrystalline powder of L-α-alanine at from 77K to 310K. The formed main radicals at 190K∼310K were the deamino radical and the decarboxyl radical. Because of the first-derivative ESR, decarboxyl radical showed an expanded spectral width and a lower peak height because of its amino hydrogen and nitrogen than that of the same amount of deamino radical. The ESR of irradiated L-α-alanine predominantly indicates the spectrum of the deamino radical. On the irradiated, L-α-alanine at from 77K to 310K ESR showed 1:4:6:4:1 lines at 220K and at room temperature, which indicate that the methyl group of the radical was rotating. On the other hand, at 77K ESR the spectrum showed nearly 1:5:5:5:1 lines, like the teeth of a saw, on samples irradiated at 270K∼350K (range IV), and 1:4:6:4:1 lines for those irradiated at 180K∼260K (range II and III), respectively. It is considered that the radical conformation of the deamino radical is planar (most stable conformation) on an irradiated sample in range IV, and a pyramidal structure on the irradiated sample in ranges II and III. (author)

A Clostridium difficile alanine racemase affects spore germination and accommodates serine as a substrate.

Science.gov (United States)

Shrestha, Ritu; Lockless, Steve W; Sorg, Joseph A

2017-06-23

Clostridium difficile has become one of the most common bacterial pathogens in hospital-acquired infections in the United States. Although C. difficile is strictly anaerobic, it survives in aerobic environments and transmits between hosts via spores. C. difficile spore germination is triggered in response to certain bile acids and glycine. Although glycine is the most effective co-germinant, other amino acids can substitute with varying efficiencies. Of these, l-alanine is an effective co-germinant and is also a germinant for most bacterial spores. Many endospore-forming bacteria embed alanine racemases into their spore coats, and these enzymes are thought to convert the l-alanine germinant into d-alanine, a spore germination inhibitor. Although the C. difficile Alr2 racemase is the sixth most highly expressed gene during C. difficile spore formation, a previous study reported that Alr2 has little to no role in germination of C. difficile spores in rich medium. Here, we hypothesized that Alr2 could affect C. difficile l-alanine-induced spore germination in a defined medium. We found that alr2 mutant spores more readily germinate in response to l-alanine as a co-germinant. Surprisingly, d-alanine also functioned as a co-germinant. Moreover, we found that Alr2 could interconvert l- and d-serine and that Alr2 bound to l- and d-serine with ∼2-fold weaker affinity to that of l- and d-alanine. Finally, we demonstrate that l- and d-serine are also co-germinants for C. difficile spores. These results suggest that C. difficile spores can respond to a diverse set of amino acid co-germinants and reveal that Alr2 can accommodate serine as a substrate. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Structural surprises in friction-deposited films of poly(tetrafluoroethylene)

DEFF Research Database (Denmark)

Breiby, Dag Werner; Sølling, Theis Ivan; Bunk, Oliver

2005-01-01

Thin films of poly(tetrafluoroethylene) (PTFE) produced by friction deposition were studied using grazing incidence X-ray diffraction as the principal tool. The structure of the deposited thin films was compared with that of the surface of the PTFE bar used for depositing the films. Both exhibited...... the 15/7 helix conformation characteristic of crystal PTFE phase IV. A high degree of biaxial orientation was found for the highly crystalline thin films. Whereas the unit cell of the bar surface material appeared to be single-stem hexagonal, the film displayed diffraction characteristics consistent...... the possibility of a continuous transition between the low-order single-stem hexagonal and the multistem high-order unit cell. The degree of chain orientation was much lower at the surface of the bar than in the thin film. A modification of the commonly accepted mechanism for the transfer of material from the bar...

Genome-wide identification and analysis of basic helix-loop-helix domains in dog, Canis lupus familiaris.

Science.gov (United States)

Wang, Xu-Hua; Wang, Yong; Liu, A-Ke; Liu, Xiao-Ting; Zhou, Yang; Yao, Qin; Chen, Ke-Ping

2015-04-01

The basic helix-loop-helix (bHLH) domain is a highly conserved amino acid motif that defines a group of DNA-binding transcription factors. bHLH proteins play essential regulatory roles in a variety of biological processes in animal, plant, and fungus. The domestic dog, Canis lupus familiaris, is a good model organism for genetic, physiological, and behavioral studies. In this study, we identified 115 putative bHLH genes in the dog genome. Based on a phylogenetic analysis, 51, 26, 14, 4, 12, and 4 dog bHLH genes were assigned to six separate groups (A-F); four bHLH genes were categorized as ''orphans''. Within-group evolutionary relationships inferred from the phylogenetic analysis were consistent with positional conservation, other conserved domains flanking the bHLH motif, and highly conserved intron/exon patterns in other vertebrates. Our analytical results confirmed the GenBank annotations of 89 dog bHLH proteins and provided information that could be used to update the annotations of the remaining 26 dog bHLH proteins. These data will provide good references for further studies on the structures and regulatory functions of bHLH proteins in the growth and development of dogs, which may help in understanding the mechanisms that underlie the physical and behavioral differences between dogs and wolves.

Serum alanine aminotransferase levels, hematocrit rate and body weight correlations before and after hemodialysis session

Directory of Open Access Journals (Sweden)

Edmundo Pessoa Lopes

2009-01-01

Full Text Available PURPOSE: To evaluate alanine aminotransferase levels before and after a hemodialysis session and to correlate these values with the hematocrit rate and weight loss during hemodialysis. PATIENTS AND METHODS: The serum alanine aminotransferase levels, hematocrit rate and body weight were measured and correlated before and after a single hemodialysis session for 146 patients with chronic renal failure. An receiver operating characteristic (ROC curve for the serum alanine aminotransferase levels collected before and after hemodialysis was plotted to identify hepatitis C virus-infected patients. RESULTS: The mean weight loss of the 146 patients during hemodialysis was 5.3% (p < 0.001. The mean alanine aminotransferase levels before and after hemodialysis were 18.8 and 23.9 IU/, respectively, denoting a significant 28.1% increase. An equally significant increase of 16.4% in the hematocrit rate also occurred after hemodialysis. The weight loss was inversely correlated with the rise in both the alanine aminotransferase level (r = 0.3; p < 0.001 and hematocrit rate (r = 0.5; p < 0.001. A direct correlation was found between the rise in alanine aminotransferase levels and the hematocrit during the hemodialysis session (r = 0.4; p < 0.001. Based on the ROC curve, the upper limit of the normal alanine aminotransferase level should be reduced by 40% relative to the upper limit of normal if the blood samples are collected before the hemodialysis session or by 60% if blood samples are collected after the session. CONCLUSION: In the present study, significant elevations in the serum alanine aminotransferase levels and hematocrit rates occurred in parallel to a reduction in body weight after the hemodialysis session. These findings suggest that one of the factors for low alanine aminotransferase levels prior to hemodialysis could be hemodilution in patients with chronic renal failure.

Effect of the ionizing radiation on alanine solution for a dosimeter application

International Nuclear Information System (INIS)

Abdessamad, Nour El Houda

2007-01-01

The amino acid alanine is well known as a dosimetric detector material for high level dosimetry. Its application is based on the formation of radicals by ionising radiation. In this study the effect of several parameters such as: the ionising radiation, the concentration, the dose on the pH, conductivity and the oscillotitrometric answer of L a lanine solution was investigated. The results show that there is a significant production of new species. The formation of these species increases upon increasing dose. The comparison between the repeatability of the used techniques led us to choose of the system alanine/pH and the alanine/conductivity as the most adapted. (Author)

Development of a dosimeter for high doses assessment based on Alanine/EPR

International Nuclear Information System (INIS)

Galante, O.L.; Rodrigues, O. Jr.; Campos, L.L.

2000-01-01

The increasing use of radiation sources of high activity for industrial and medical applications becomes important the research and the development of detectors and dosimetric methods for quality control of the applied doses. This work presents the current stage of the research at IPEN/CNEN-SP that has as objective the development of a standard dosimetric system for high doses assessment based on the alanine as radiation detector and electron paramagnetic resonance (EPR) as measurement technique. The developed system consists of the cylindrical container built in polyethylene of high density and the detector element based on DL-alanine commercially available. For the detector preparation different binding materials such as paraffin and acetate polyvinyl solution (pva) and also the use of a polyethylene tube of low density with 3.2 mm of external diameter, 2 mm of internal diameter and 30 mm of length were tested to provide the easier preparation method and the most sensitive detector. For the alanine + paraffin detector it was used 80% of alanine and 20% of paraffin, for the alanine + pva detector it was used 70% of alanine and 30% of pva solution, and pure alanine was encapsulated, compacted and sealed in the case of the polyethylene tube. The obtained results with respect to handling, packing and construction easiness showed that the polyethylene tube presents all characteristics to obtain of a good detector element. The validation of the dosimetric system was carried out with gamma radiation of the cobalt-60 with doses in the range between 0.2 Gy to 200 kGy. Type tests such as fading, lowest detection limit, reproducibility and energy dependence of the sign EPR were performed. All measurements were carried out at room temperature using a spectrometer of electron paramagnetic resonance (EPR) Bruker model MXE. Taking into account the results obtained: linearity of the EPR signal between 10 Gy and 50 kGy, reproducibility better than 3%, low fading associated with

The influence of measurement and storage conditions on alanine ESR dosimeters

International Nuclear Information System (INIS)

Alexandre, A.C.

1992-01-01

Alanine has several desirable properties as an ESR dosemeter e.g. tissue equivalence, low fading and an approximately linear response for doses up to 10 kGy. This work reports on a simple system to produce the alanine dosemeter, the signal intensity for a range of doses and energies, and the effect of the air humidity and the spectrometer settings on the ESR signal. (Author)

Tah1 helix-swap dimerization prevents mixed Hsp90 co-chaperone complexes

International Nuclear Information System (INIS)

Morgan, Rhodri M. L.; Pal, Mohinder; Roe, S. Mark; Pearl, Laurence H.; Prodromou, Chrisostomos

2015-01-01

A helix swap involving the fifth helix between two adjacently bound Tah1 molecules restores the normal binding environment of the conserved MEEVD peptide of Hsp90. Dimerization also explains how other monomeric TPR-domain proteins are excluded from forming inappropriate mixed co-chaperone complexes with Hsp90 and Tah1. Specific co-chaperone adaptors facilitate the recruitment of client proteins to the Hsp90 system. Tah1 binds the C-terminal conserved MEEVD motif of Hsp90, thus linking an eclectic set of client proteins to the R2TP complex for their assembly and regulation by Hsp90. Rather than the normal complement of seven α-helices seen in other tetratricopeptide repeat (TPR) domains, Tah1 unusually consists of the first five only. Consequently, the methionine of the MEEVD peptide remains exposed to solvent when bound by Tah1. In solution Tah1 appears to be predominantly monomeric, and recent structures have failed to explain how Tah1 appears to prevent the formation of mixed TPR domain-containing complexes such as Cpr6–(Hsp90) 2 –Tah1. To understand this further, the crystal structure of Tah1 in complex with the MEEVD peptide of Hsp90 was determined, which shows a helix swap involving the fifth α-helix between two adjacently bound Tah1 molecules. Dimerization of Tah1 restores the normal binding environment of the bound Hsp90 methionine residue by reconstituting a TPR binding site similar to that in seven-helix-containing TPR domain proteins. Dimerization also explains how other monomeric TPR-domain proteins are excluded from forming inappropriate mixed co-chaperone complexes

Tah1 helix-swap dimerization prevents mixed Hsp90 co-chaperone complexes

Energy Technology Data Exchange (ETDEWEB)

Morgan, Rhodri M. L.; Pal, Mohinder; Roe, S. Mark; Pearl, Laurence H., E-mail: laurence.pearl@sussex.ac.uk; Prodromou, Chrisostomos, E-mail: laurence.pearl@sussex.ac.uk [University of Sussex, Falmer, Brighton BN1 9RQ (United Kingdom)

2015-05-01

A helix swap involving the fifth helix between two adjacently bound Tah1 molecules restores the normal binding environment of the conserved MEEVD peptide of Hsp90. Dimerization also explains how other monomeric TPR-domain proteins are excluded from forming inappropriate mixed co-chaperone complexes with Hsp90 and Tah1. Specific co-chaperone adaptors facilitate the recruitment of client proteins to the Hsp90 system. Tah1 binds the C-terminal conserved MEEVD motif of Hsp90, thus linking an eclectic set of client proteins to the R2TP complex for their assembly and regulation by Hsp90. Rather than the normal complement of seven α-helices seen in other tetratricopeptide repeat (TPR) domains, Tah1 unusually consists of the first five only. Consequently, the methionine of the MEEVD peptide remains exposed to solvent when bound by Tah1. In solution Tah1 appears to be predominantly monomeric, and recent structures have failed to explain how Tah1 appears to prevent the formation of mixed TPR domain-containing complexes such as Cpr6–(Hsp90){sub 2}–Tah1. To understand this further, the crystal structure of Tah1 in complex with the MEEVD peptide of Hsp90 was determined, which shows a helix swap involving the fifth α-helix between two adjacently bound Tah1 molecules. Dimerization of Tah1 restores the normal binding environment of the bound Hsp90 methionine residue by reconstituting a TPR binding site similar to that in seven-helix-containing TPR domain proteins. Dimerization also explains how other monomeric TPR-domain proteins are excluded from forming inappropriate mixed co-chaperone complexes.

Fluorophores, environments, and quantification techniques in the analysis of transmembrane helix interaction using FRET.

Science.gov (United States)

Khadria, Ambalika S; Senes, Alessandro

2015-07-01

Förster resonance energy transfer (FRET) has been widely used as a spectroscopic tool in vitro to study the interactions between transmembrane (TM) helices in detergent and lipid environments. This technique has been instrumental to many studies that have greatly contributed to quantitative understanding of the physical principles that govern helix-helix interactions in the membrane. These studies have also improved our understanding of the biological role of oligomerization in membrane proteins. In this review, we focus on the combinations of fluorophores used, the membrane mimetic environments, and measurement techniques that have been applied to study model systems as well as biological oligomeric complexes in vitro. We highlight the different formalisms used to calculate FRET efficiency and the challenges associated with accurate quantification. The goal is to provide the reader with a comparative summary of the relevant literature for planning and designing FRET experiments aimed at measuring TM helix-helix associations. © 2015 Wiley Periodicals, Inc.

Calculations of the relative effectiveness of alanine for neutrons with energies up to 17.1 MeV

International Nuclear Information System (INIS)

Gerstenberg, H.M.; Coyne, J.J.

1990-01-01

The relative effectiveness (RE) of alanine has been calculated for neutrons using the RE of alanine for charged particles. The neutrons interact with one or more of the elements (hydrogen, carbon, nitrogen and oxygen) that compose the alanine. These interactions produce spectra of secondary charged particles consisting of ions of H, D, He, Be, B, C, N and O. From a combination of the calculated secondary charged particle spectra generated by the slowing down neutrons, and the calculated RE of the ions produced, a RE for the neutrons can be obtained. In addition, lineal energy spectra were determined for neutrons with energies up to 17.1 MeV interacting with alanine. An analytical code was used to calculate these spectra for a 1 μm diameter alanine cell surrounded by an alanine medium. For comparison, similar calculations were made for muscle tissue. Finally, the calculated differential RE was folded with dose distributions to obtain RE-weighted distributions for alanine. (author)

Glucose-stimulated prehepatic insulin secretion is associated with circulating alanine, triglyceride, glucagons, lactate and TNF-alfa in patients with HIV-lipodystrophy

DEFF Research Database (Denmark)

Haugaard, Steen B; Andersen, Ove; Pedersen, SB

2006-01-01

with the remaining HIV-infected patients (all Ptriglyceride, alanine, glucagon, lactate and TNF-alpha may be associated with alterations in the first-phase prehepatic insulin secretion response to intravenous glucose in normoglycaemic lipodystrophic HIV-infected patients.......OBJECTIVES: We examined whether insulin-resistant lipodystrophic HIV-infected patients with known high fasting prehepatic insulin secretion rates (FISRs) displayed alterations in first-phase prehepatic insulin response to intravenous glucose (ISREG0-10 min). METHODS: Eighteen normoglycaemic...... lipodystrophic HIV-infected (LIPO) patients and 25 normoglycaemic nonlipodystrophic HIV-infected patients (controls) were included in the study. The prehepatic insulin secretion rate was estimated by deconvolution of C-peptide concentrations, and insulin sensitivity (SIRd) was estimated by the glucose clamp...

Effects of dose fractionation on the response of alanine dosimetry

International Nuclear Information System (INIS)

Lundahl, Brad; Logar, John; Desrosiers, Marc; Puhl, James

2014-01-01

Alanine dosimetry is well established as a transfer standard and is becoming more prevalently used in routine dosimetry systems for radiation processing. Many routine measurement applications in radiation processing involve absorbed dose measurements resulting from fractioned exposures to ionizing radiation. Fractioning of absorbed dose is identified as an influence quantity (ISO/ASTM, 2013). This paper reports on study results of absorbed dose fractioning characteristics of alanine for gamma and high energy electron beam radiation sources. The results of this study indicate a radiation response difference due to absorbed dose fractioning in response can be observed after four fractionations for high-energy electron beams and no difference up to seven fractions for gamma rays using an ANOVA evaluation method. - Highlights: • Fractioning effects signaled in electron beam using an ANOVA at 6 equal increments. • Fractioning effects not signaled in gamma using an ANOVA up to 7 equal increments. • Insensitivity of alanine to dose fractioning indicates nominal impact on calibration

Determination of the carbon, hydrogen and nitrogen contents of alanine and their uncertainties using the certified reference material L-alanine (NMIJ CRM 6011-a).

Science.gov (United States)

Itoh, Nobuyasu; Sato, Ayako; Yamazaki, Taichi; Numata, Masahiko; Takatsu, Akiko

2013-01-01

The carbon, hydrogen, and nitrogen (CHN) contents of alanine and their uncertainties were estimated using a CHN analyzer and the certified reference material (CRM) L-alanine. The CHN contents and their uncertainties, as measured using the single-point calibration method, were 40.36 ± 0.20% for C, 7.86 ± 0.13% for H, and 15.66 ± 0.09% for N; the results obtained using the bracket calibration method were also comparable. The method described in this study is reasonable, convenient, and meets the general requirement of having uncertainties ≤ 0.4%.

Kinetic and modelling studies of NAD+ and poly(ethylene glycol)-bound NAD+ in horse liver alcohol dehydrogenase

NARCIS (Netherlands)

Vanhommerig, S.A.M.; Sluyterman, L.A.A.E.; Meijer, E.M.

1996-01-01

Poly(ethylene glycol)-bound nicotinamide adenine dinucleotide (PEG-NAD+) has been successfully employed in the continuous production of L-amino acids from the corresponding alpha-keto acids by stereospecific reductive amination. Like many other dehydrogenases also horse liver alcohol dehydrogenase

Dependence of alanine gel dosimeter response as a function of photon clinical beams dose rate

International Nuclear Information System (INIS)

Silva, Cleber Feijo; Campos, Leticia Lucente

2013-01-01

Gel dosimetry is a new area developed by Gore, it is ery useful for application in radiotherapy because using NMR imaging as evaluation technique is possible to evaluate three dimensional absorbed dose distribution. The measure technique is based on difference of ferrous (Fe 2+ ) and ferric (Fe 3+ ) ) ions concentration that can be measured also by spectrophotometry technique. The Alanine gel dosimeter was developed at IPEN. The alanine is an amino acid and tissue equivalent material that presents significant improvement on previous alanine dosimetry systems. The addition of Alanine increases the production of ferric ions in the solution. This work aims to study the dose rate dependence of photon clinical beams radiation on the alanine gel dosimeter optical response, as well as the response repeatability and gel production reproducibility, since this property is very important for characterization and standardization of any dosimeter. (author)

The photon energy dependence of the alanine/EPR dosimetry system, an experimental investigation

International Nuclear Information System (INIS)

Bergstrand, E.S.; Hole, E.O.; Shortt, K.R.; Ross, C.K.

2002-01-01

The energy dependence of a dosimetry system based on electron paramagnetic resonance (EPR) spectroscopy of alanine has been studied to determine its suitability for use in dose verification for radiotherapy. A few experiments with high-energy photon irradiation of alanine have been reported in the literature. However, the reported results disagree whether the ratio of dose in alanine to dose in water is independent of the radiation energy or whether there is a small dependence for photon energies of relevance to radiotherapy. The concentration of free radicals in alanine is proportional to the absorbed dose in alanine over a wide dose range covering three decades. The relative number of radicals may be determined by examining the EPR spectrum, and hence it is possible to determine the dose with a system that has been calibrated using a known dose of 60 Co radiation. In the present work, irradiations of alanine dosimeters were performed at the National Research Council (NRC), in Ottawa, Canada. The radiation qualities investigated were 10, 20 and 30 MV x-rays using the NRC linac. For each radiation quality, 30 dosimeters were irradiated in a water phantom with a level of absorbed dose to water ranging from 10 to 50 Gy. For reference purposes, irradiations using the NRC 60 Co source were performed on more or less the same day as the irradiations at each specific linac quality. In all beams, the dose to water was measured using a graphite-walled NE2571 ionisation chamber that was originally calibrated by comparison with a sealed-water calorimeter. The alanine dosimeters were evaluated at the EPR laboratory at the University of Oslo, Norway, using an X-band Bruker ESP300E spectrometer with a rectangular double resonator. One of the resonators contained a Mn 2+ /MgO sample that was read after each dosimeter reading, in order to provide independence from short-term sensitivity fluctuations in the spectrometer. All dosimeters irradiated at one specific linac quality were

An investigation of the photon energy dependence of the EPR alanine dosimetry system

International Nuclear Information System (INIS)

Bergstrand, Eva Stabell; Shortt, Ken R; Ross, Carl K; Hole, Eli Olaug

2003-01-01

The electron paramagnetic resonance (EPR) alanine dosimetry system is based on EPR measurements of radicals formed in alanine by ionizing radiation. The system has been studied to determine its energy dependence for photons in the 10-30 MV region relative to those of 60 Co and to find out if the system would be suitable for dosimetry comparisons. The irradiations were carried out at the National Research Council, Ottawa, Canada and the doses ranged from 8 to 54 Gy. The EPR measurements were performed at the University of Oslo, Norway. The ratio of the slope of the alanine reading versus dose-to-water curve for a certain linac photon beam quality and the corresponding slope for a reference 60 Co γ-radiation gives an experimental measure of the relative dose-to-water response of the EPR alanine dosimetry system. For calculating the linear regression coefficients of these alanine reading versus dose curves, the method of weighted least squares was used. This method is assumed to produce more accurate regression coefficients when applied to EPR dosimetry than the common method of standard least squares. The overall uncertainty on the ratio of slopes was between 0.5 and 0.6% for all three linac energies. The relative response for all the linac beams compared to cobalt was less than unity: by about 0.5% for the 20 and 30 MV points but by more than 1% for the 10 MV point. The given standard uncertainties negate concluding that there is any significant internal variation in the measured response as a function of beam quality between the three linac energies. Thus, we calculated the average dose response for all three energies and found that the alanine response is 0.8% (±0.5%) lower for high energy x-rays than for 60 Co γ-rays. This result indicates a small energy dependence in the alanine response for the high-energy photons relative to 60 Co which may be significant. This result is specific to our dosimetry system (alanine with 20% polyethylene binder pressed into a

Atomistic model of the spider silk nanostructure

Science.gov (United States)

Keten, Sinan; Buehler, Markus J.

2010-04-01

Spider silk is an ultrastrong and extensible self-assembling biopolymer that outperforms the mechanical characteristics of many synthetic materials including steel. Here we report atomic-level structures that represent aggregates of MaSp1 proteins from the N. Clavipes silk sequence based on a bottom-up computational approach using replica exchange molecular dynamics. We discover that poly-alanine regions predominantly form distinct and orderly beta-sheet crystal domains while disorderly structures are formed by poly-glycine repeats, resembling 31-helices. These could be the molecular source of the large semicrystalline fraction observed in silks, and also form the basis of the so-called "prestretched" molecular configuration. Our structures are validated against experimental data based on dihedral angle pair calculations presented in Ramachandran plots, alpha-carbon atomic distances, as well as secondary structure content.

Spectrophotometric readout for alanine dosimeter to be used for food irradiation applications

International Nuclear Information System (INIS)

Ebraheem, S.; Beshir, W.B.; Sobhy, R.; Kovacs, A.; Wojnarovits, L.

2002-01-01

Complete text of publication follows. The electron spin resonance (EPR) readout of radical concentration produced upon irradiation of L-alanine is well known as a transfer dosimetry system for high dose range. The high cost of EPR/alanine dosimetry system is a serious handicap for large-scale routine application for use in irradiation facilities. In this study the irradiated L-alanine was dissolved in 1,4 -phenyl diammonium dichloride (PAC) solution resulting a complex of a purple colour. This complex has a variable absorbance with applied dose in the range from 1 - 20 kGy. The absorbance of this complex using spectrophotometric evaluation was investigated at 401 and 478 nm with increasing dose. The fluorimetric emission was also studied at the wavelength of 475 nm as a function of dose. This methos for dose evaluation is uncomplicated and very easy for routine application. The effect of the dye concentration as well as the suitable amount of alanine has been studied with respect to practical use. The stability of the complex for long period of time was also investigated

Selfcalibrated alanine/EPR dosimeters. A new generation of solid state/EPR dosimeters

International Nuclear Information System (INIS)

Yordanov, N.D.; Gancheva, V.

1999-01-01

Alanine/EPR dosimeters are well established as secondary, reference dosimeters for high-energy radiation. However, there are various sources of uncertainty in the evaluation of absorbed dose. This arises primarily from the necessity to calibrate each EPR spectrometer and each batch of dosimeters before their use. In order to overcome this disadvantage, a new generation alanine/EPR dosimeter has been developed, and its possibilities as a radiation detector are reported. Principally, it is a mixture of alanine, some quantity of EPR active substance, and a binding material. The EPR active substance, acting as an internal EPR standard, is chosen to have EPR parameters which are independent of the irradiation dose. The simultaneous recording of the spectra of both the sample and the standard under the same experimental conditions and the estimation of the ratio I alanine /I Mn as a function of the absorbed dose strongly reduces the uncertainties. The response of these dosimeters for 60 Co γ-radiation exhibits excellent linearity and reproducibility in the range of absorbed dose, 10 2 - 5 x 10 4 Gy. (author)

Influence of the irradiation temperature on the free-radical response of alanine

International Nuclear Information System (INIS)

Wieser, A.; Siegele, R.; Regulla, D.F.

1989-01-01

GSF operates the only IAEA high-level dosimetry reference laboratory and, as a joint project with the IAEA, the International Dose Assurance Service (IDAS). Dosimetry is based on long-lived free radicals in organic alanine induced by ionizing radiation and readout by ESR spectroscopy. The thermal time response of the radical concentration in alanine is fairly constant after irradiation provided that the alanine samples are stored at temperatures below 50 0 C. By contrast, a positive temperature coefficient had earlier been found at GSF for the production rate of alanine radicals, for irradiation temperatures between 0 and 50 0 C. This effect has to be considered for reference dosimetry in radiation processing. Radiation processing is also of interest at irradiation temperatures below 0 0 C. The present study describes experiments on the influence of irradiation temperatures between +50 and -100 0 C. Comparison is made between the present and earlier results, in the overlapping temperature range. An empirical function is proposed for the temperature coefficient based on the experimental data. (author)

Gate-controlled switching between persistent and inverse persistent spin helix states

International Nuclear Information System (INIS)

Yoshizumi, K.; Sasaki, A.; Kohda, M.; Nitta, J.

2016-01-01

We demonstrate gate-controlled switching between persistent spin helix (PSH) state and inverse PSH state, which are detected by quantum interference effect on magneto-conductance. These special symmetric spin states showing weak localization effect give rise to a long spin coherence when the strength of Rashba spin-orbit interaction (SOI) is close to that of Dresselhaus SOI. Furthermore, in the middle of two persistent spin helix states, where the Rashba SOI can be negligible, the bulk Dresselhaus SOI parameter in a modulation doped InGaAs/InAlAs quantum well is determined.

Gate-controlled switching between persistent and inverse persistent spin helix states

Energy Technology Data Exchange (ETDEWEB)

Yoshizumi, K.; Sasaki, A.; Kohda, M.; Nitta, J. [Department of Materials Science, Tohoku University, Sendai 980-8579 (Japan)

2016-03-28

We demonstrate gate-controlled switching between persistent spin helix (PSH) state and inverse PSH state, which are detected by quantum interference effect on magneto-conductance. These special symmetric spin states showing weak localization effect give rise to a long spin coherence when the strength of Rashba spin-orbit interaction (SOI) is close to that of Dresselhaus SOI. Furthermore, in the middle of two persistent spin helix states, where the Rashba SOI can be negligible, the bulk Dresselhaus SOI parameter in a modulation doped InGaAs/InAlAs quantum well is determined.

The Quadruple Helix Model Enhancing Innovative Performance Of Indonesian Creative Industry

Directory of Open Access Journals (Sweden)

Sri Wahyu Lelly Hana Setyanti

2017-11-01

Full Text Available The creative industry in Indonesia has contributed positively to the national economic growth. Creative industry grows from the creativity and innovation performance of the business actors. The challenge of creative industry is how to completely understand the creative and innovative processes in business management. Therefore it requires an approach that combines the synergy between academicians entrepreneurs government and society in a quadruple helix model. The objective of this research is to develop a creativity model through a quadruple helix model in improving innovation performance of the creative industry.

Design and synthesis of DNA four-helix bundles

Energy Technology Data Exchange (ETDEWEB)

Rangnekar, Abhijit; Gothelf, Kurt V [Department of Chemistry, Centre for DNA Nanotechnology (CDNA) and Interdisciplinary Nanoscience Center (iNANO), Aarhus University, DK-8000 Aarhus C (Denmark); LaBean, Thomas H, E-mail: kvg@chem.au.dk, E-mail: thl@cs.duke.edu [Department of Chemistry, Duke University, Durham, NC 27708 (United States)

2011-06-10

The field of DNA nanotechnology has evolved significantly in the past decade. Researchers have succeeded in synthesizing tile-based structures and using them to form periodic lattices in one, two and three dimensions. Origami-based structures have also been used to create nanoscale structures in two and three dimensions. Design and construction of DNA bundles with fixed circumference has added a new dimension to the field. Here we report the design and synthesis of a DNA four-helix bundle. It was found to be extremely rigid and stable. When several such bundles were assembled using appropriate sticky-ends, they formed micrometre-long filaments. However, when creation of two-dimensional sheet-like arrays of the four-helix bundles was attempted, nanoscale rings were observed instead. The exact reason behind the nanoring formation is yet to be ascertained, but it provides an exciting prospect for making programmable circular nanostructures using DNA.

Design and synthesis of DNA four-helix bundles

International Nuclear Information System (INIS)

Rangnekar, Abhijit; Gothelf, Kurt V; LaBean, Thomas H

2011-01-01

The field of DNA nanotechnology has evolved significantly in the past decade. Researchers have succeeded in synthesizing tile-based structures and using them to form periodic lattices in one, two and three dimensions. Origami-based structures have also been used to create nanoscale structures in two and three dimensions. Design and construction of DNA bundles with fixed circumference has added a new dimension to the field. Here we report the design and synthesis of a DNA four-helix bundle. It was found to be extremely rigid and stable. When several such bundles were assembled using appropriate sticky-ends, they formed micrometre-long filaments. However, when creation of two-dimensional sheet-like arrays of the four-helix bundles was attempted, nanoscale rings were observed instead. The exact reason behind the nanoring formation is yet to be ascertained, but it provides an exciting prospect for making programmable circular nanostructures using DNA.

Different β-alanine dimeric forms in trifluoromethanesulfonic acid salts. XRD and vibrational studies

Science.gov (United States)

Wołoszyn, Łukasz; Ilczyszyn, Maria M.

2018-03-01

Two new crystalline salts: β-alaninium trifluoromethanesulfonate (β-AlaOTf) and bis(β-alanine) trifluoromethanesulfonate (β-2AlaOTf) were obtained. The former one contains diprotonated β-alanine dimer, the latter one monoprotonated β-alanine dimer. Both compounds were studied by single crystal XRD, vibrational (IR and Raman) spectroscopy and calorimetric method. The quantum-mechanical calculations (DFT/B3LYP/6-311 ++G(2d,2p)) for the diprotonated dimer were carried out. The β-AlaOTf salt crystallizes in the P 1 bar space group of triclinic system (Z = 2), the β-2AlaOTf in the P21/m space group of monoclinic system (Z = 2). The vibrational data for the studied compounds are discussed in relation to their crystal structure, and provide insight into the character of hydrogen bonds and β-alanine protonation. The studied crystals do not exhibit phase transitions in the solid state.

Potential energy surface of alanine polypeptide chains

DEFF Research Database (Denmark)

Solov'yov, Ilia; Yakubovich, Alexander V.; Solov'yov, Andrey V.

2006-01-01

The multidimensional potential energy surfaces of the peptide chains consisting of three and six alanine (Ala) residues have been studied with respect to the degrees of freedom related to the twist of these molecules relative to the peptide backbone (these degrees of freedom are responsible...

Absorbed dose determination in kilovoltage X-ray synchrotron radiation using alanine dosimeters.

Science.gov (United States)

Butler, D J; Lye, J E; Wright, T E; Crossley, D; Sharpe, P H G; Stevenson, A W; Livingstone, J; Crosbie, J C

2016-12-01

Alanine dosimeters from the National Physical Laboratory (NPL) in the UK were irradiated using kilovoltage synchrotron radiation at the imaging and medical beam line (IMBL) at the Australian Synchrotron. A 20 × 20 mm 2 area was irradiated by scanning the phantom containing the alanine through the 1 mm × 20 mm beam at a constant velocity. The polychromatic beam had an average energy of 95 keV and nominal absorbed dose to water rate of 250 Gy/s. The absorbed dose to water in the solid water phantom was first determined using a PTW Model 31014 PinPoint ionization chamber traceable to a graphite calorimeter. The alanine was read out at NPL using correction factors determined for 60 Co, traceable to NPL standards, and a published energy correction was applied to correct for the effect of the synchrotron beam quality. The ratio of the doses determined by alanine at NPL and those determined at the synchrotron was 0.975 (standard uncertainty 0.042) when alanine energy correction factors published by Waldeland et al. (Waldeland E, Hole E O, Sagstuen E and Malinen E, Med. Phys. 2010, 37, 3569) were used, and 0.996 (standard uncertainty 0.031) when factors by Anton et al. (Anton M, Büermann L., Phys Med Biol. 2015 60 6113-29) were used. The results provide additional verification of the IMBL dosimetry.

Dose Response of Alanine Detectors Irradiated with Carbon Ion Beams

DEFF Research Database (Denmark)

Herrmann, Rochus; Jäkel, Oliver; Palmans, Hugo

2011-01-01

Purpose: The dose response of the alanine detector shows a dependence on particle energy and type, when irradiated with ion beams. The purpose of this study is to investigate the response behaviour of the alanine detector in clinical carbon ion beams and compare the results with model predictions......-dose curves deviate from predictions in the peak region, most pronounced at the distal edge of the peak. Conclusions: The used model and its implementation show a good overall agreement for quasi mono energetic measurements. Deviations in depth-dose measurements are mainly attributed to uncertainties...

Spatial attributes of the four-helix bundle group of bacteriocins – The high-resolution structure of BacSp222 in solution

KAUST Repository

Nowakowski, Michał

2017-11-01

BacSp222 is a multifunctional bacteriocin produced by Staphylococcus pseudintermedius strain 222, an opportunistic pathogen of domestic animals. At micromolar concentrations, BacSp222 kills Gram-positive bacteria and is cytotoxic toward mammalian cells, while at nanomolar doses, it acts as an immunomodulatory factor, enhancing nitric oxide release in macrophage-like cell lines. The bacteriocin is a cationic, N-terminally formylated, 50-amino-acid-long linear peptide that is rich in tryptophan residues.In this study, the solution structure of BacSp222 was determined and compared to the currently known structures of similar bacteriocins. BacSp222 was isolated from a liquid culture medium in a uniformly 13C- and 15N-labeled form, and NMR data were collected. The structure was calculated based on NMR-derived constraints and consists of a rigid and tightly packed globular bundle of four alpha-helices separated by three short turns.Although the amino acid sequence of BacSp222 has no significant similarity to any known peptide or protein, a 3D structure similarity search indicates a close relation to other four-helix bundle-motif bacteriocins, such as aureocin A53, lacticin Q and enterocins 7A/7B. Assuming similar functions, biology, structure and physicochemical properties, we propose to distinguish the four-helix bundle bacteriocins as a new Type A in subclass IId of bacteriocins, containing linear, non-pediocin-like peptides.

Green reduction of graphene oxide using alanine.

Science.gov (United States)

Wang, Jiabin; Salihi, Elif Caliskan; Šiller, Lidija

2017-03-01

There remains a real need for the easy, eco-friendly and scalable preparation method of graphene due to various potential applications. Chemical reduction is the most versatile method for the large scale production of graphene. Here we report the operating conditions for a one-step, economical and green synthesis method for the reduction of graphene oxide using a biomolecule (alanine). Graphene oxide was produced by the oxidation and exfoliation of natural graphite flake with strong oxidants using Hummers method (Hummers and Offeman, 1958), but the method was revised in our laboratory to set up a safe and environmentally friendly route. The reduction of graphene oxide was investigated using alanine at various operating conditions in order to set up optimum conditions (treatment time, temperature and concentration of the reagent). Samples have been characterized by using UV-Visible spectroscopy, Fourier transform infrared spectroscopy, transmission electron microscopy, Raman spectroscopy and X-ray diffraction analysis. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

Alanine-EPR dosimetry system. Why we like it?

International Nuclear Information System (INIS)

Stuglik, Z.

2007-01-01

To develop a new high-dose dosimeter we should: (1) to find material with radiation effect monotonically (if possible linearly) dependent on an absorbed dose; (2) to investigate its dosimetric characteristics (sensitivity, dose range, repeatability, accuracy, post-effects); (3) to evaluate economical parameters of new method (cost and availability of dosimetric material, cost of analytical instrument and its services); (4) to evaluate operational features of new dosimeter (sensitivity for environmental conditions, time from irradiation to the read-out); (5) to perform a calibration curve, i.e. functional dependence between radiation effect (dosimetric signal) and absorbed dose. On the base of this very stable stable ammonium radical (SAR) generated in crystalline α-alanine was established in the INCT as an alanine-EPR dosimetry system. Presented lecture describes the main features of this dosimeter

A method to improve application technique in Alanine/ESR dosimetry

International Nuclear Information System (INIS)

Choi, Hoon; Ha, Ju Hee; Choi, Won; Lim, Young Khi

2008-01-01

Full text: For long-term radiation monitoring to assess the cable aging in harsh condition of nuclear power plant, ESR dosimetry method using alanine dosimeters was already recommended in many technical document and paper. Several ESR dosimetry systems were already produced and used widely, but the actual application of these systems for industrial dosimetry needs careful consideration of error sources in process of dose measurement. The alanine dosimeters were measured by E-scan alanine analyzer system or EMX ESR spectrometer. For the accurate measurement of the dosimeters, we have studied various source of errors. First, this paper discusses sources of inaccuracy related to data processing. To make a meaningful dose assessment, the dosimeter measurements need to be compared with measurements made using certified dosimeters of known dose. This is achieved by performing the routine calibration procedure which creates a calibration curve and corresponding fit coefficients from measurements made with a set of dosimeters with known certified dose. The calibration curves in ESR dosimetry are usually constructed by means of the least-squares technique in its simplest variant. The recommended alternative linearity several replicate measurements of Y at each used X value. Also, measurement is subject to error, so repeat measurements will not be identical. A technical description of a linear calibration is assumed that the dispersion of the measurements is the same for each standard. But in some cases, the standard deviation has to be specified separately for each value of concentration. In this case, it used the WLS (Weighted Least-Squared Regression) method instead of the OLS (Ordinary Least-Squared Regression) method. Second, the precision of the ESR spectrum was showed as a reproducibility test for the two ESR systems. The reproducibility test was performed at an absorbed dose of 1 k Gy, which is at least three orders of magnitude above the background reading of an

Different β-alanine dimeric forms in trifluoromethanesulfonic acid salts. XRD and vibrational studies.

Science.gov (United States)

Wołoszyn, Łukasz; Ilczyszyn, Maria M

2018-03-15

Two new crystalline salts: β-alaninium trifluoromethanesulfonate (β-AlaOTf) and bis(β-alanine) trifluoromethanesulfonate (β-2AlaOTf) were obtained. The former one contains diprotonated β-alanine dimer, the latter one monoprotonated β-alanine dimer. Both compounds were studied by single crystal XRD, vibrational (IR and Raman) spectroscopy and calorimetric method. The quantum-mechanical calculations (DFT/B3LYP/6-311++G(2d,2p)) for the diprotonated dimer were carried out. The β-AlaOTf salt crystallizes in the P1¯ space group of triclinic system (Z=2), the β-2AlaOTf in the P2 1 /m space group of monoclinic system (Z=2). The vibrational data for the studied compounds are discussed in relation to their crystal structure, and provide insight into the character of hydrogen bonds and β-alanine protonation. The studied crystals do not exhibit phase transitions in the solid state. Copyright © 2017 Elsevier B.V. All rights reserved.

Electron paramagnetic resonance radiation dosimetry: possible inorganic alternatives to the EPR/alanine dosimeter

International Nuclear Information System (INIS)

Keizer, P.N.; Morton, J.R.; Preston, K.F.

1991-01-01

The intensity of the EPR spectrum of γ-irradiated L-α-alanine has been accepted by the International Atomic Energy Agency as a secondary standard for high-dose (10-100 000 Gy) dosimetry. The alanine dosimeter is not without its disadvantages, however, and in this article alternative EPR dosimeters are explored. These include SO 3 - in irradiated K 2 CH 2 (SO 3 ) 2 and CO 2 - in irradiated sodium formate (NaHCO 2 ), both of which have some advantages over CH 3 CHCO 2 - in L-α-alanine. Using as a readout parameter the peak-to-peak excursion of the strongest line, these systems have a four-fold sensitivity advantage over alanine. The radicals SO 3 - and CO 2 - are, moreover, found in a wide variety of matrices, and it may be possible to find one in which they are even stronger. The need to discover a dosimeter material sensitive enough to function in the 'clinical' dose range (below 10 Gy) is emphasized. (author)

Knockout of the alanine racemase gene in Lactobacillus plantarum results in septation defects and cell wall perforation

NARCIS (Netherlands)

Palumbo, E.; Favier, C.F.; Deghorain, M.; Cocconcelli, P.S.; Grangette, C.; Mercenier, A.M.E.; Vaughan, E.E.; Hols, P.

2004-01-01

A stable mutant of Lactobacillus plantarum deficient in alanine racemase (Alr) was constructed by two successive homologous recombination steps. When the mutant was supplemented with D-alanine, growth and viability were unaffected. Surprisingly, deprivation Of D-alanine during exponential growth did

Effect of poly and mono-unsaturated fatty acids on stability and structure of recombinant S100A8/A9.

Science.gov (United States)

Asghari, Hamideh; Chegini, Koorosh Goodarzvand; Amini, Abbas; Gheibi, Nematollah

2016-03-01

Recombinant pET 15b vectors containing the coding sequences S100A8 and S100A9 are expressed in Escherichia coli BL21 (DE3) and purified using Ni-NTA affinity chromatography. The structural changes of S100A8/A9 complex are analyzed upon interaction with poly/mono-unsaturated fatty acids (UFAs). The thermodynamic values, Gibbs free energy and the protein melting point, are obtained through thermal denaturation of protein both with and without UFAs by thermal scanning of protein emission using the fluorescence spectroscopy technique. The far-ultraviolet circular dichroism spectra show that all studied unsaturated fatty acids, including arachidonic, linoleic, alpha-linolenic and oleic acids, induce changes in the secondary structure of S100A8/A9 by reducing the α-helix and β-sheet structures. The tertiary structure of S100A8/A9 has fluctuations in the fluorescence emission spectra after the incubation of protein with UFAs. The blue-shift of emission maximum wavelength and the increase in fluorescence intensity of anilino naphthalene-8-sulfonic acid confirm that the partial unfolding is caused by the conformational changes in the tertiary structure in the presence of UFAs. The structural changes in S100A8/A9 and its lower stability in the presence of UFAs may be necessary for S100A8/A9 to play a biological role in the inflammatory milieu. Copyright © 2015 Elsevier B.V. All rights reserved.

Preliminary assessment of LiF and alanine detectors for the dosimetry of proton therapy beams

International Nuclear Information System (INIS)

Fattibene, P.; Calicchia, A.; De Angelis, C.; Onori, S.; Egger, E.

1996-01-01

An experimental intercomparison between the proton response of LiF TLD-100 and alanine detectors is reported. The investigations were performed with LiF chips and alanine pellets in a 62 MeV proton beam at the Paul Scherrer Institut in Villigen (CH). Results were compared with reference dosimetry provided by Markus type parallel plate ionization chamber. The response of the detectors was studied, in a phantom, at different beam penetration depths in pristine and modulated beams. For both alanine and TL detectors, within the experimental uncertainty of the measurements, no significant energy dependence in the response was observed down to the Bragg peak region. The sensitivity of alanine and LiF detectors to protons was measured in the centre of modulated Bragg peak and no significant difference was found with respect to 60 Co. Contrary to LiF, alanine also offers a remarkable tissue equivalence which favours its choice for in-phantom dosimetry. (author)

Sorption and Microbial Uptake of Alanine, Glucose and Acetate in Soil

Science.gov (United States)

Fischer, H.; Ingwersen, J.; Kuzyakov, Y.

2009-04-01

Low molecular weight organic substances (LMWOS), e. g. amino acids, sugars, and carboxylic acids, are C compounds that are most rapidly turned-over in the C cycle of soil. Despite of their importance it is still unknown how sorption to the soil matrix affects their turnover in soil solution. The goals of this study were (1) to describe the dynamics of the fluxes of LMWOS (10 µmol l-1) in various pools (dissolved, adsorbed, decomposed to CO2, incorporated into microbial biomass) and (2) to assess the LMWOS distribution in these pools in dependence of very wide range of concentration (0.01 to 1000 µmol l-1). Representatives of each LMWOS group (glucose for sugars, alanine for amino acids, Na-acetate for carboxylic acids) uniformly labeled with 14C were added to sterilized or non-sterilized soil and analyzed in dif-ferent compartments between 1 min and 5.6 hours after addition. LMWOS were almost completely taken up by microorganisms within the first 30 min. Microbial uptake was much faster than the physicochemical sorption (estimated in sterilized soil), which needed to reach quasi-equilibrium 60 min for alanine and about 400 min for glucose. Only sorption of acetate was instantaneous (>1 min). While for acetate the maximum sorption capacity was reached at 100 µmol l-1 no such maximum was found for glucose and alanine in the studied concentra-tion range. At the concentration of 100 µmol l-1, microbial decomposition after 4.5 h hours was higher for alanine (76.7±1.1%) than acetate (55.2±0.9%) and glucose (28.5±1.5%). On the contrary, incorporation into microbial biomass was higher for glucose (59.8±1.2%) than for acetate (23.4±5.9%) and alanine (5.2±2.8%). Within 10 to 500 µmol l-1 the pathways of the three LMWOS transformation changed: at 500 µmol l-1 alanine and acetate were less mineralized and more incorporated into microbial biomass than at 10 µmol l-1, while glucose incorporation decreased. Consequently, the concentrations of alanine, glucose, and

Interfacial electrostatics of poly(vinylamine hydrochloride), poly(diallyldimethylammonium chloride), poly-l-lysine, and poly-l-arginine interacting with lipid bilayers.

Science.gov (United States)

McGeachy, A C; Dalchand, N; Caudill, E R; Li, T; Doğangün, M; Olenick, L L; Chang, H; Pedersen, J A; Geiger, F M

2018-04-25

Charge densities of cationic polymers adsorbed to lipid bilayers are estimated from second harmonic generation (SHG) spectroscopy and quartz crystal microbalance with dissipation monitoring (QCM-D) measurements. The systems surveyed included poly(vinylamine hydrochloride) (PVAm), poly(diallyldimethylammonium chloride) (PDADMAC), poly-l-lysine (PLL), and poly-l-arginine (PLR), as well as polyalcohol controls. Upon accounting for the number of positive charges associated with each polyelectrolyte, the binding constants and apparent free energies of adsorption as estimated from SHG data are comparable despite differences in molecular masses and molecular structure, with ΔGads values of -61 ± 2, -58 ± 2, -57 ± 1, -52 ± 2, -52 ± 1 kJ mol-1 for PDADMAC400, PDADMAC100, PVAm, PLL, and PLR, respectively. Moreover, we find charge densities for polymer adlayers of approximately 0.3 C m-2 for poly(diallyldimethylammonium chloride) while those of poly(vinylamine) hydrochloride, poly-l-lysine, and poly-l-arginine are approximately 0.2 C m-2. Time-dependent studies indicate that polycation adsorption to supported lipid bilayers is only partially reversible for most of the polymers explored. Poly(diallyldimethylammonium chloride) does not demonstrate reversible binding even over long timescales (>8 hours).

Dosimetric calibration of humidity chamber inside the 60Co-PANBIT irradiator using alanine ESR dosimeter

International Nuclear Information System (INIS)

Murali, S.; Venkataramani, R.; Pushparaja; Natarajan, V.; Sastry, M.D.; Bora, J.S.; Venkatacharyulu, K.

1998-01-01

In the present work the suitability of the electron spin resonance (ESR) technique, using DL - α - alanine sample is examined. It is well documented that radiation induced free radicals in alanine give strong ESR spectrum which can be used for dosimetry purposes. The aspects that are relevant to the present work are: (I) stability of the radicals in temperature and humidity conditions in which the experiments were carried out; (II) linearity at high doses; and (III) establishing the utility of alanine ESR dosimeter by cross checking the dose values with more established dosimeters. The details of these investigations are presented and it is shown that alanine ESR dosimeter will meet all the required conditions satisfactorily

Mechanism of death at high temperatures in Helix and Patella

Energy Technology Data Exchange (ETDEWEB)

Grainger, J N.R.

1975-10-01

In Patella vulgata and Helix aspersa which had been killed by exposure to high temperatures, the rates of oxygen consumption of gill, foot muscle and hepatopancreas are remarkably steady when measured at lower temperatures, although the absolute levels are in some cases different from normal animals. These tissues are thus substantially metabolically intact in heat dead individuals. In Helix there is a fall in blood sodium and a rise in blood potassium during heat death. In Patella there is a marked rise in blood Na/sup +/ and a consequent disturbance of the Na/sup +//K/sup +/ ratio. These ionic disturbances are thought to be a prime cause of heat death. The significance of the results is discussed.

Alanine dosimetry using a spectrophotometric ferric-xylenol orange complex readout

International Nuclear Information System (INIS)

Laere, K. van; Buysse, J.; Berkvens, P.

1989-01-01

The spectrophotometric dosimetric method using the indirect oxidation of ferrous ions after dissolution of irradiated DL-and L-alanine has been thoroughly investigated with respect to its composition, read-out procedure and dose-response. Optimal concentration of 0.10 N H 2 SO 4 , 0.2 mM xylenol orange and 0.2 mM Fe 2+ were found, giving an absorption maximum at 547 nm. Standardization of chemical processing procedures allows a reproducibility better than 0.5%. The useful dose range has been extended to 0.03-12 kGy by means of slightly different read-out procedure. The quantitative concept of ''indirect yield'', G id , was introduced for this procedure as a measure of the indirect oxidation capacity of the radicals. It was found to be G id,0 (Fe 3+ ) 7.1 ions/100 eV transferred into the alanine. The spectrophotometric readout combines the highly advantageous use of alanine as a dosemeter with the straightforwardness, accuracy and low costs of the chemical procedure. (author)

[Establishing biological reference intervals of alanine transaminase for clinical laboratory stored database].

Science.gov (United States)

Guo, Wei; Song, Binbin; Shen, Junfei; Wu, Jiong; Zhang, Chunyan; Wang, Beili; Pan, Baishen

2015-08-25

To establish an indirect reference interval based on the test results of alanine aminotransferase stored in a laboratory information system. All alanine aminotransferase results were included for outpatients and physical examinations that were stored in the laboratory information system of Zhongshan Hospital during 2014. The original data were transformed using a Box-Cox transformation to obtain an approximate normal distribution. Outliers were identified and omitted using the Chauvenet and Tukey methods. The indirect reference intervals were obtained by simultaneously applying nonparametric and Hoffmann methods. The reference change value was selected to determine the statistical significance of the observed differences between the calculated and published reference intervals. The indirect reference intervals for alanine aminotransferase of all groups were 12 to 41 U/L (male, outpatient), 12 to 48 U/L (male, physical examination), 9 to 32 U/L (female, outpatient), and 8 to 35 U/L (female, physical examination), respectively. The absolute differences when compared with the direct results were all smaller than the reference change value of alanine aminotransferase. The Box-Cox transformation combined with the Hoffmann and Tukey methods is a simple and reliable technique that should be promoted and used by clinical laboratories.

ESR investigation of L-α-alanine and sucrose radicals produced by heavy-ion irradiation

International Nuclear Information System (INIS)

Nakagawa, K.; Sato, Y.

2005-01-01

We investigated sucrose and L-α-alanine radicals produced by heavy (particle) ion irradiation with various LETs (linear energy transfer). The impact of the heavy ions on the samples produced stable free radicals, which were analyzed by ESR (electron spin resonance). Identical spectra were measured after one year. The obtained spectral patterns were the same as those for helium (He), carbon (C), and neon (Ne) ions irradiation. The absorbed dose dependences for the irradiated sucrose and alanine samples were examined. The ESR response has a linear relation with the absorbed dose. The ESR response at 60 Gy was slightly lower than a linear line for sucrose; however, the response showed good linearity for the alanine. In addition, the total spin concentration obtained by heavy-ion irradiation correlated logarithmically with the LET. Qualitative ESR analyse showed that the production of sucrose and alanine radicals depended on both different particle irradiation and the LET under the same dose. Thus, the present ESR results imply that sucrose together with L-α-alanine can be used to monitor LET as well as the number of ionizing particle for the production of stable free radicals. (author)

Thermochemistry of the solution of β-alanine in (H2O + alcohol) mixtures at 298.15 K

International Nuclear Information System (INIS)

Smirnov, Valeriy I.; Badelin, Valentin G.

2013-01-01

Highlights: • Enthalpies of β-alanine dissolution have been measured in aqueous solution of MeOH, EtOH, 1-PrOH and 2-PrOH. • Measured data were reported as functions of composition of water + alcohol mixtures. • Enthalpy coefficients of pairwise interactions have been analyzed in terms of McMillan–Mayer theory. - Abstract: The enthalpies of the solution of β-alanine in H 2 O + (methanol, ethanol, 1-propanol and 2-propanol) mixtures with alcohol content up to 0.4 mol fractions, have been determined calorimetrically at T = 298.15 K. The standard enthalpies of the solution and transfer of β-alanine from water to aqueous alcohol have been calculated. The effect of structure properties of a mixed solvent on specified enthalpy characteristics of β-alanine is discussed. The enthalpy coefficients of pairwise interactions between β-alanine and alcohol molecules have been computed. It has been found that these coefficients become increasingly positive in methanol, ethanol, 1-propanol, and 2-propanol sequence. A comparative analysis of thermodynamic characteristics of dissolution of β-alanine and D,L-α-alanine in the mixtures studied has been made

De novo design, synthesis and characterisation of MP3, a new catalytic four-helix bundle hemeprotein.

Science.gov (United States)

Faiella, Marina; Maglio, Ornella; Nastri, Flavia; Lombardi, Angela; Lista, Liliana; Hagen, Wilfred R; Pavone, Vincenzo

2012-12-07

A new artificial metalloenzyme, MP3 (MiniPeroxidase 3), designed by combining the excellent structural properties of four-helix bundle protein scaffolds with the activity of natural peroxidases, was synthesised and characterised. This new hemeprotein model was developed by covalently linking the deuteroporphyrin to two peptide chains of different compositions to obtain an asymmetric helix-loop-helix/heme/helix-loop-helix sandwich arrangement, characterised by 1) a His residue on one chain that acts as an axial ligand to the iron ion; 2) a vacant distal site that is able to accommodate exogenous ligands or substrates; and 3) an Arg residue in the distal site that should assist in hydrogen peroxide activation to give an HRP-like catalytic process. MP3 was synthesised and characterised as its iron complex. CD measurements revealed the high helix-forming propensity of the peptide, confirming the appropriateness of the model procedure; UV/Vis, MCD and EPR experiments gave insights into the coordination geometry and the spin state of the metal. Kinetic experiments showed that Fe(III)-MP3 possesses peroxidase-like activity comparable to R38A-hHRP, highlighting the possibility of mimicking the functional features of natural enzymes. The synergistic application of de novo design methods, synthetic procedures, and spectroscopic characterisation, described herein, demonstrates a method by which to implement and optimise catalytic activity for an enzyme mimetic. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Structural plasticity of the N-terminal capping helix of the TPR domain of kinesin light chain.

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The Quyen Nguyen

Full Text Available Kinesin1 plays a major role in neuronal transport by recruiting many different cargos through its kinesin light chain (KLC. Various structurally unrelated cargos interact with the conserved tetratricopeptide repeat (TPR domain of KLC. The N-terminal capping helix of the TPR domain exhibits an atypical sequence and structural features that may contribute to the versatility of the TPR domain to bind different cargos. We determined crystal structures of the TPR domain of both KLC1 and KLC2 encompassing the N-terminal capping helix and show that this helix exhibits two distinct and defined orientations relative to the rest of the TPR domain. Such a difference in orientation gives rise, at the N-terminal part of the groove, to the formation of one hydrophobic pocket, as well as to electrostatic variations at the groove surface. We present a comprehensive structural analysis of available KLC1/2-TPR domain structures that highlights that ligand binding into the groove can be specific of one or the other N-terminal capping helix orientations. Further, structural analysis reveals that the N-terminal capping helix is always involved in crystal packing contacts, especially in a TPR1:TPR1' contact which highlights its propensity to be a protein-protein interaction site. Together, these results underline that the structural plasticity of the N-terminal capping helix might represent a structural determinant for TPR domain structural versatility in cargo binding.

Comparison of the UDP-N-Acetylmuramate:l-Alanine Ligase Enzymes from Mycobacterium tuberculosis and Mycobacterium leprae

OpenAIRE

Mahapatra, Sebabrata; Crick, Dean C.; Brennan, Patrick J.

2000-01-01

In the peptidoglycan of Mycobacterium leprae, l-alanine of the side chain is replaced by glycine. When expressed in Escherichia coli, MurC (UDP-N-acetyl-muramate:l-alanine ligase) of M. leprae showed Km and Vmax for l-alanine and glycine similar to those of Mycobacterium tuberculosis MurC, suggesting that another explanation should be sought for the presence of glycine.

Effect of the ionizing radiation on alanine solution for a dosimeter application

International Nuclear Information System (INIS)

Ketata, Ameni

2011-01-01

The electron spin resonance spectroscopy is well known as a reference and transfer dosimetry system for the evaluation of high doses in radiation processing. The high cost of an EPR/alanine dosimetry system is a serious handicap for large-scale routine application. In this study, the use of irradiated L-alanine dissolved in color indicator solutions (bromothymol blue and fuchsin) was investigated for dosimetry purposes. This solution has an absorbance varies linearly with the absorbed dose in the dose range of 0-25 kGy for the bromothymol blue, and 0-45 kGy for the fuchsin. The effects of the dye and the alanine concentration, the p H value as well as of the solvent have been studied. With respect to routine application, the stability of dosimeters was also investigated

Controlling chirality with helix inversion in cholesteric liquid crystals

NARCIS (Netherlands)

Katsonis, Nathalie Hélène; Lacaze, E.; Ferrarini, A.

2012-01-01

The helical organization of cholesteric liquid crystals is omnipresent in living matter. Achieving control over the structure of the cholesteric helix consequently holds great potential for developing stimuli-responsive materials matching the level of sophistication of biological systems. In

Accurate measurement of the optical activity of alanine crystals and the determination of their absolute chirality

Science.gov (United States)

Ishikawa, Kazuhiko; Terasawa, Yukana; Tanaka, Masahito; Asahi, Toru

2017-05-01

Wavelength dependence measurements of the chiroptical properties in alanine crystals have so far been unsuccessful using conventional spectroscopic techniques. We describe our attempts to measure the wavelength dependence of the optical activity in L- and D-alanine crystals along each crystallographic axis, and to determine the absolute chirality of alanine crystals by correlating the absolute structure to the optical activity using an x-ray diffractometer and a generalized high accuracy universal polarimeter. We have succeeded in accurately measuring the optical rotatory dispersion in the direction, which shows that the optical rotation of the D-alanine crystal is dextrorotatory and that of the L-alanine crystal is laevorotatory, thereby determining the absolute chirality. Furthermore, comparison with the optical activity in solution shows that the optical activity in alanine crystals is different not only in value, but also in the sign. These results have led us to conclude that the optical rotatory power in the crystalline state should not be simply the summation of molecular optical rotatory power values. We propose the necessity of a theory, which contains the contribution of molecular interactions within the crystal, in order to calculate the optical rotatory power of the crystalline state.

Dosimetry auditing procedure with alanine dosimeters for light ion beam therapy

DEFF Research Database (Denmark)

Ableitinger, Alexander; Vatnitsky, Stanislav; Herrmann, Rochus

2013-01-01

Background and purpose In the next few years the number of facilities providing ion beam therapy with scanning beams will increase. An auditing process based on an end-to-end test (including CT imaging, planning and dose delivery) could help new ion therapy centres to validate their entire logistic...... of the biological dose is out of scope of the current work. Materials and methods The audit procedure was based on a homogeneous phantom that mimics the dimension of a head (20 × 20 × 21 cm3). The phantom can be loaded either with an ionisation chamber or 20 alanine dosimeters plus 2 radiochromic EBT films. Dose...... fluence in the alanine detector. A pilot run was performed with protons and carbon ions at the Heidelberg Ion Therapy facility (HIT). Results The mean difference of the absolute physical dose measured with the alanine dosimeters compared with the expected dose from the treatment planning system was −2...

Alanine and TLD coupled detectors for fast neutron dose measurements in neutron capture therapy (NCT)

Energy Technology Data Exchange (ETDEWEB)

Cecilia, A.; Baccaro, S.; Cemmi, A. [ENEA-FIS-ION, Casaccia RC, Via Anguillarese 301, 00060 Santa Maria di Galeria, Rome (Italy); Colli, V.; Gambarini, G. [Dept. of Physics of the Univ., INFN, Via Celoria 16, 20133 Milan (Italy); Rosi, G. [ENEA-FIS-ION, Casaccia RC, Via Anguillarese 301, 00060 Santa Maria di Galeria, Rome (Italy); Scolari, L. [Dept. of Physics of the Univ., INFN, Via Celoria 16, 20133 Milan (Italy)

2004-07-01

A method was investigated to measure gamma and fast neutron doses in phantoms exposed to an epithermal neutron beam designed for neutron capture therapy (NCT). The gamma dose component was measured by TLD-300 [CaF{sub 2}:Tm] and the fast neutron dose, mainly due to elastic scattering with hydrogen nuclei, was measured by alanine dosemeters [CH{sub 3}CH(NH{sub 2})COOH]. The gamma and fast neutron doses deposited in alanine dosemeters are very near to those released in tissue, because of the alanine tissue equivalence. Couples of TLD-300 and alanine dosemeters were irradiated in phantoms positioned in the epithermal column of the Tapiro reactor (ENEA-Casaccia RC). The dosemeter response depends on the linear energy transfer (LET) of radiation, hence the precision and reliability of the fast neutron dose values obtained with the proposed method have been investigated. Results showed that the combination of alanine and TLD detectors is a promising method to separate gamma dose and fast neutron dose in NCT. (authors)

Thermal decomposition behavior of potassium and sodium jarosite synthesized in the presence of methylamine and alanine

Energy Technology Data Exchange (ETDEWEB)

J. Michelle Kotler; Nancy W. Hinman; C. Doc Richardson; Jill R. Scott

2010-10-01

Biomolecules, methylamine and alanine, found associated with natural jarosite samples peaked the interest of astrobiologists and planetary geologists. How the biomolecules are associated with jarosite remains unclear although the mechanism could be important for detecting biosignatures in the rock record on Earth and other planets. A series of thermal gravimetric experiments using synthetic K-jarosite and Na-jarosite were conducted to determine if thermal analysis could differentiate physical mixtures of alanine and methylamine with jarosite from samples where the methylamine or alanine was incorporated into the synthesis procedure. Physical mixtures and synthetic experiments with methylamine and alanine could be differentiated from one another and from the standards by thermal analysis for both the K-jarosite and Na-jarosite end-member suites. Changes included shifts in on-set temperatures, total temperature changes from on-set to final, and the presence of indicator peaks for methylamine and alanine in the physical mixture experiments.

Equilibrium shift in solution: molecular shape recognition and precipitation of a synthetic double helix using helicene-grafted silica nanoparticles.

Science.gov (United States)

Miyagawa, Masamichi; Ichinose, Wataru; Yamaguchi, Masahiko

2014-01-27

Chiral silica nanoparticles (70 nm) grafted with (P)-helicene recognized the molecular shape of double helix and random coil (P)-ethynylhelicene oligomers in solution. A mixture of the (P)-nanoparticles and double helix precipitated much faster than a mixture of the (P)-nanoparticles and random coil, and the precipitate contained only the double helix. The mixture of the (P)-nanoparticles and (P)-ethynylhelicene pentamer reversibly dispersed in trifluoromethylbenzene upon heating at 70 °C and precipitated upon cooling at 25 °C. When a 10:90 equilibrium mixture of the double helix and random coil in solution was treated with the (P)-nanoparticles, the double helix was precipitated in 53% yield and was accompanied by equilibrium shift. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Classification and evolutionary analysis of the basic helix-loop-helix gene family in the green anole lizard, Anolis carolinensis.

Science.gov (United States)

Liu, Ake; Wang, Yong; Zhang, Debao; Wang, Xuhua; Song, Huifang; Dang, Chunwang; Yao, Qin; Chen, Keping

2013-08-01

Helix-loop-helix (bHLH) proteins play essential regulatory roles in a variety of biological processes. These highly conserved proteins form a large transcription factor superfamily, and are commonly identified in large numbers within animal, plant, and fungal genomes. The bHLH domain has been well studied in many animal species, but has not yet been characterized in non-avian reptiles. In this study, we identified 102 putative bHLH genes in the genome of the green anole lizard, Anolis carolinensis. Based on phylogenetic analysis, these genes were classified into 43 families, with 43, 24, 16, 3, 10, and 3 members assigned into groups A, B, C, D, E, and F, respectively, and 3 members categorized as "orphans". Within-group evolutionary relationships inferred from the phylogenetic analysis were consistent with highly conserved patterns observed for introns and additional domains. Results from phylogenetic analysis of the H/E(spl) family suggest that genome and tandem gene duplications have contributed to this family's expansion. Our classification and evolutionary analysis has provided insights into the evolutionary diversification of animal bHLH genes, and should aid future studies on bHLH protein regulation of key growth and developmental processes.

Structure of alanine racemase from Oenococcus oeni with bound pyridoxal 5′-phosphate

International Nuclear Information System (INIS)

Palani, Kandavelu; Burley, Stephen K.; Swaminathan, Subramanyam

2012-01-01

Alanine racemase from O. oeni exists as a dimer in the crystal structure. Both monomers contribute to the two active sites present, one for each monomer. The crystal structure of alanine racemase from Oenococcus oeni has been determined at 1.7 Å resolution using the single-wavelength anomalous dispersion (SAD) method and selenium-labelled protein. The protein exists as a symmetric dimer in the crystal, with both protomers contributing to the two active sites. Pyridoxal 5′-phosphate, a cofactor, is bound to each monomer and forms a Schiff base with Lys39. Structural comparison of alanine racemase from O. oeni (Alr) with homologous family members revealed similar domain organization and cofactor binding

Comparison of the UDP-N-Acetylmuramate:l-Alanine Ligase Enzymes from Mycobacterium tuberculosis and Mycobacterium leprae

Science.gov (United States)

Mahapatra, Sebabrata; Crick, Dean C.; Brennan, Patrick J.

2000-01-01

In the peptidoglycan of Mycobacterium leprae, l-alanine of the side chain is replaced by glycine. When expressed in Escherichia coli, MurC (UDP-N-acetyl-muramate:l-alanine ligase) of M. leprae showed Km and Vmax for l-alanine and glycine similar to those of Mycobacterium tuberculosis MurC, suggesting that another explanation should be sought for the presence of glycine. PMID:11073931

The metabolism of histamine in the Drosophila optic lobe involves an ommatidial pathway: β-alanine recycles through the retina

Science.gov (United States)

Borycz, Janusz; Borycz, Jolanta A.; Edwards, Tara N.; Boulianne, Gabrielle L.; Meinertzhagen, Ian A.

2012-01-01

SUMMARY Flies recycle the photoreceptor neurotransmitter histamine by conjugating it to β-alanine to form β-alanyl-histamine (carcinine). The conjugation is regulated by Ebony, while Tan hydrolyses carcinine, releasing histamine and β-alanine. In Drosophila, β-alanine synthesis occurs either from uracil or from the decarboxylation of aspartate but detailed roles for the enzymes responsible remain unclear. Immunohistochemically detected β-alanine is present throughout the fly’s entire brain, and is enhanced in the retina especially in the pseudocone, pigment and photoreceptor cells of the ommatidia. HPLC determinations reveal 10.7 ng of β-alanine in the wild-type head, roughly five times more than histamine. When wild-type flies drink uracil their head β-alanine increases more than after drinking l-aspartic acid, indicating the effectiveness of the uracil pathway. Mutants of black, which lack aspartate decarboxylase, cannot synthesize β-alanine from l-aspartate but can still synthesize it efficiently from uracil. Our findings demonstrate a novel function for pigment cells, which not only screen ommatidia from stray light but also store and transport β-alanine and carcinine. This role is consistent with a β-alanine-dependent histamine recycling pathway occurring not only in the photoreceptor terminals in the lamina neuropile, where carcinine occurs in marginal glia, but vertically via a long pathway that involves the retina. The lamina’s marginal glia are also a hub involved in the storage and/or disposal of carcinine and β-alanine. PMID:22442379

Designer interface peptide grafts target estrogen receptor alpha dimerization

International Nuclear Information System (INIS)

Chakraborty, S.; Asare, B.K.; Biswas, P.K.; Rajnarayanan, R.V.

2016-01-01

The nuclear transcription factor estrogen receptor alpha (ERα), triggered by its cognate ligand estrogen, regulates a variety of cellular signaling events. ERα is expressed in 70% of breast cancers and is a widely validated target for anti-breast cancer drug discovery. Administration of anti-estrogen to block estrogen receptor activation is still a viable anti-breast cancer treatment option but anti-estrogen resistance has been a significant bottle-neck. Dimerization of estrogen receptor is required for ER activation. Blocking ERα dimerization is therefore a complementary and alternative strategy to combat anti-estrogen resistance. Dimer interface peptide “I-box” derived from ER residues 503–518 specifically blocks ER dimerization. Recently using a comprehensive molecular simulation we studied the interaction dynamics of ERα LBDs in a homo-dimer. Based on this study, we identified three interface recognition peptide motifs LDKITDT (ERα residues 479–485), LQQQHQRLAQ (residues 497–506), and LSHIRHMSNK (residues 511–520) and reported the suitability of using LQQQHQRLAQ (ER 497–506) as a template to design inhibitors of ERα dimerization. Stability and self-aggregation of peptide based therapeutics poses a significant bottle-neck to proceed further. In this study utilizing peptide grafted to preserve their pharmacophoric recognition motif and assessed their stability and potential to block ERα mediated activity in silico and in vitro. The Grafted peptides blocked ERα mediated cell proliferation and viability of breast cancer cells but did not alter their apoptotic fate. We believe the structural clues identified in this study can be used to identify novel peptidometics and small molecules that specifically target ER dimer interface generating a new breed of anti-cancer agents. - Highlights: • Designer peptide grafts retain core molecular recognition motif during MD simulations. • Designer peptide grafts with Poly-ALA helix form stable

Designer interface peptide grafts target estrogen receptor alpha dimerization

Energy Technology Data Exchange (ETDEWEB)

Chakraborty, S. [Laboratory of Computational Biophysics & Bioengineering, Department of Physics, Tougaloo College, Tougaloo, MS 39174 (United States); Asare, B.K. [Department of Pharmacology and Toxicology, University of Buffalo, Buffalo, NY 14214 (United States); Biswas, P.K., E-mail: pbiswas@tougaloo.edu [Laboratory of Computational Biophysics & Bioengineering, Department of Physics, Tougaloo College, Tougaloo, MS 39174 (United States); Rajnarayanan, R.V., E-mail: rajendra@buffalo.edu [Department of Pharmacology and Toxicology, University of Buffalo, Buffalo, NY 14214 (United States)

2016-09-09

The nuclear transcription factor estrogen receptor alpha (ERα), triggered by its cognate ligand estrogen, regulates a variety of cellular signaling events. ERα is expressed in 70% of breast cancers and is a widely validated target for anti-breast cancer drug discovery. Administration of anti-estrogen to block estrogen receptor activation is still a viable anti-breast cancer treatment option but anti-estrogen resistance has been a significant bottle-neck. Dimerization of estrogen receptor is required for ER activation. Blocking ERα dimerization is therefore a complementary and alternative strategy to combat anti-estrogen resistance. Dimer interface peptide “I-box” derived from ER residues 503–518 specifically blocks ER dimerization. Recently using a comprehensive molecular simulation we studied the interaction dynamics of ERα LBDs in a homo-dimer. Based on this study, we identified three interface recognition peptide motifs LDKITDT (ERα residues 479–485), LQQQHQRLAQ (residues 497–506), and LSHIRHMSNK (residues 511–520) and reported the suitability of using LQQQHQRLAQ (ER 497–506) as a template to design inhibitors of ERα dimerization. Stability and self-aggregation of peptide based therapeutics poses a significant bottle-neck to proceed further. In this study utilizing peptide grafted to preserve their pharmacophoric recognition motif and assessed their stability and potential to block ERα mediated activity in silico and in vitro. The Grafted peptides blocked ERα mediated cell proliferation and viability of breast cancer cells but did not alter their apoptotic fate. We believe the structural clues identified in this study can be used to identify novel peptidometics and small molecules that specifically target ER dimer interface generating a new breed of anti-cancer agents. - Highlights: • Designer peptide grafts retain core molecular recognition motif during MD simulations. • Designer peptide grafts with Poly-ALA helix form stable

Purification, characterization and gene cloning of thermostable O-acetyl-L-serine sulfhydrylase forming β-cyano-L-alanine

International Nuclear Information System (INIS)

Omura, Hironori; Yoshida, Toyokazu; Nagasawa, Toru; Kobayashi, Michihiko; Shimizu, Sakayu

2003-01-01

A thermophilic and cyanide ion-tolerant bacterium, Bacillus stearothermophilus CN3 isolated from a hot spring in Japan, was found to produce thermostable β-cyano-L-alanine synthase. The enzyme catalyzes the synthesis of β-cyano-L-alanine from O-acetyl-L-serine and cyanide ions. The purified enzyme has a molecular mass of approximately 70 kDa and consists of two identical sub-units. It was stable in the pH range of 6.0 to 10.0 and up to 70degC. The enzyme also catalyzes the synthesis of various β-substituted-L-alanine derivatives from O-acetyl-L-serine and nucleophilic reagents. The gene encoding the β-cyano-L-alanine synthase was isolated from B. stearothermophilus CN3. Sequence homology analysis revealed that the β-cyano-L-alanine synthase of the bacterium is O-acetyl-L-serine sulfhydrylase. A recombinant plasmid, constructed by ligation of the cloned gene and an expression vector, pKK223-3, was introduced into E. coli JM109. The transformed E. coli cells overexpressed β-cyano-L-alanine synthase. Heat stable β-cyano-L-alanine synthase can be applied to the synthesis of [4- 11 C]L-2,4-diaminobutyric acid as a tracer for positron emission tomography. (author)

Purification, characterization and gene cloning of thermostable O-acetyl-L-serine sulfhydrylase forming {beta}-cyano-L-alanine

Energy Technology Data Exchange (ETDEWEB)

Omura, Hironori; Yoshida, Toyokazu; Nagasawa, Toru [Gifu Univ. (Japan). Dept. of Biomolecular Science; Kuroda, Masako [Ikeda Food Research Co., Ltd., Fukuyama, Hiroshima (Japan); Kobayashi, Michihiko; Shimizu, Sakayu [Kyoto Univ. (Japan). Agricultural Sciences

2003-10-01

A thermophilic and cyanide ion-tolerant bacterium, Bacillus stearothermophilus CN3 isolated from a hot spring in Japan, was found to produce thermostable {beta}-cyano-L-alanine synthase. The enzyme catalyzes the synthesis of {beta}-cyano-L-alanine from O-acetyl-L-serine and cyanide ions. The purified enzyme has a molecular mass of approximately 70 kDa and consists of two identical sub-units. It was stable in the pH range of 6.0 to 10.0 and up to 70degC. The enzyme also catalyzes the synthesis of various {beta}-substituted-L-alanine derivatives from O-acetyl-L-serine and nucleophilic reagents. The gene encoding the {beta}-cyano-L-alanine synthase was isolated from B. stearothermophilus CN3. Sequence homology analysis revealed that the {beta}-cyano-L-alanine synthase of the bacterium is O-acetyl-L-serine sulfhydrylase. A recombinant plasmid, constructed by ligation of the cloned gene and an expression vector, pKK223-3, was introduced into E. coli JM109. The transformed E. coli cells overexpressed {beta}-cyano-L-alanine synthase. Heat stable {beta}-cyano-L-alanine synthase can be applied to the synthesis of [4-{sup 11}C]L-2,4-diaminobutyric acid as a tracer for positron emission tomography. (author)

Gating at the mouth of the acetylcholine receptor channel: energetic consequences of mutations in the alphaM2-cap.

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Pallavi A Bafna

2008-06-01

Full Text Available Gating of nicotinic acetylcholine receptors from a C(losed to an O(pen conformation is the initial event in the postsynaptic signaling cascade at the vertebrate nerve-muscle junction. Studies of receptor structure and function show that many residues in this large, five-subunit membrane protein contribute to the energy difference between C and O. Of special interest are amino acids located at the two transmitter binding sites and in the narrow region of the channel, where CO gating motions generate a lowhigh change in the affinity for agonists and in the ionic conductance, respectively. We have measured the energy changes and relative timing of gating movements for residues that lie between these two locations, in the C-terminus of the pore-lining M2 helix of the alpha subunit ('alphaM2-cap'. This region contains a binding site for non-competitive inhibitors and a charged ring that influences the conductance of the open pore. alphaM2-cap mutations have large effects on gating but much smaller effects on agonist binding, channel conductance, channel block and desensitization. Three alphaM2-cap residues (alphaI260, alphaP265 and alphaS268 appear to move at the outset of channel-opening, about at the same time as those at the transmitter binding site. The results suggest that the alphaM2-cap changes its secondary structure to link gating motions in the extracellular domain with those in the channel that regulate ionic conductance.

Phylogeny, Functional Annotation, and Protein Interaction Network Analyses of the Xenopus tropicalis Basic Helix-Loop-Helix Transcription Factors

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Wuyi Liu

2013-01-01

Full Text Available The previous survey identified 70 basic helix-loop-helix (bHLH proteins, but it was proved to be incomplete, and the functional information and regulatory networks of frog bHLH transcription factors were not fully known. Therefore, we conducted an updated genome-wide survey in the Xenopus tropicalis genome project databases and identified 105 bHLH sequences. Among the retrieved 105 sequences, phylogenetic analyses revealed that 103 bHLH proteins belonged to 43 families or subfamilies with 46, 26, 11, 3, 15, and 4 members in the corresponding supergroups. Next, gene ontology (GO enrichment analyses showed 65 significant GO annotations of biological processes and molecular functions and KEGG pathways counted in frequency. To explore the functional pathways, regulatory gene networks, and/or related gene groups coding for Xenopus tropicalis bHLH proteins, the identified bHLH genes were put into the databases KOBAS and STRING to get the signaling information of pathways and protein interaction networks according to available public databases and known protein interactions. From the genome annotation and pathway analysis using KOBAS, we identified 16 pathways in the Xenopus tropicalis genome. From the STRING interaction analysis, 68 hub proteins were identified, and many hub proteins created a tight network or a functional module within the protein families.

A spectrophotometric readout for γ irradiated alanine solution - a dosimetric application

International Nuclear Information System (INIS)

Marzouk, Asma

2007-01-01

Alanine is a stable dosimeter of reference in its solid state. Its installation in solution as being a dosimetric system of routine remains very useful. A follow-up of the behaviour of the irradiated alanine solution with 15 kGy according to the concentration is carried out by UV-Visible spectrophotometry. The results obtained prove the difficulty in analytical studies of the radiolysis of aqueous solutions by optical absorption due to the ambiguous broad spectra of the species and the reaction products. (Author). 47 refs

Observation of helix associations for insertion of a retinal molecule and distortions of helix structures in bacteriorhodopsin

Science.gov (United States)

Urano, Ryo; Okamoto, Yuko

2015-12-01

We applied a newly proposed prediction method for membrane protein structures to bacteriorhodopsin that has distorted transmembrane helices in the native structure. This method uses an implicit membrane model, which restricts sampling space during folding in a membrane region, and includes helix bending. Replica-exchange simulations were performed with seven transmembrane helices only without a retinal molecule. Obtained structures were classified into clusters of similar structures, which correspond to local-minimum free energy states. The two lowest free energy states corresponded to a native-like structure with the correct empty space for retinal and a structure with this empty space filled with a helix. Previous experiments of bacteriorhodopsin suggested that association of transmembrane helices enables them to make a room for insertion of a retinal. Our results are consistent with these results. Moreover, distortions of helices in the native-like structures were successfully reproduced. In the distortions, whereas the locations of kinks for all helices were similar to those of Protein Data Bank's data, the amount of bends was more similar for helices away from the retinal than for those close to the retinal in the native structure. This suggests a hypothesis that the amino-acid sequence specifies the location of kinks in transmembrane helices and that the amount of distortions depends on the interactions with the surrounding molecules such as neighboring helices, lipids, and retinal.

Identification and elucidation of in vivo function of two alanine racemases from Pseudomonas putida KT2440.

Science.gov (United States)

Duque, Estrella; Daddaoua, Abdelali; Cordero, Baldo F; De la Torre, Jesús; Antonia Molina-Henares, Maria; Ramos, Juan-Luis

2017-10-01

The genome of Pseudomonas putida KT2440 contains two open reading frames (ORFs), PP_3722 and PP_5269, that encode proteins with a Pyridoxal phosphate binding motif and a high similarity to alanine racemases. Alanine racemases play a key role in the biosynthesis of D-alanine, a crucial amino acid in the peptidoglycan layer. For these ORFs, we generated single and double mutants and found that inactivation of PP_5269 resulted in D-alanine auxotrophy, while inactivation of PP_3722 did not. Furthermore, as expected, the PP_3722/PP_5269 double mutant was a strict auxotroph for D-alanine. These results indicate that PP_5269 is an alr allele and that it is the essential alanine racemase in P. putida. We observed that the PP_5269 mutant grew very slowly, while the double PP_5269/PP_3722 mutant did not grow at all. This suggests that PP_3722 may replace PP_5269 in vivo. In fact, when the ORF encoding PP_3772 was cloned into a wide host range expression vector, ORF PP_3722 successfully complemented P. putida PP_5269 mutants. We purified both proteins to homogeneity and while they exhibit similar K M values, the V max of PP_5269 is fourfold higher than that of PP_3722. Here, we propose that PP_5269 and PP_3722 encode functional alanine racemases and that these genes be named alr-1 and alr-2 respectively. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

Swift heavy ion irradiation effects in {alpha} poly(vinylidene fluoride); Etude des effets induits par les ions lourds energetiques dans le poly(fluorure de vinylidene)

Energy Technology Data Exchange (ETDEWEB)

Le Bouedec, A

1999-11-29

The goal of this study is to characteristic and to localised defects created in {alpha} Poly (vinylidene fluoride) after swift heavy ion irradiations. PVDF films are irradiated with several Swift Heavy Ions (SHI), in the electronic stopping power (dE/dx){sub e}, in order to study the influence of irradiation parameters (absorbed dose, ion). These irradiated films are studied by different analysis techniques such as FTIR, ESR (X and Q band) spectroscopies and DSC. The crystalline level of PVDF is about 50% and we follow it destruction and amorphization as the absorbed dose increase by DSC and FTIR studies. The variation of the various FTIR bands allow us to observe the unsaturations induced by SHI radiations. Two sets of defects are observed: those which yield is sensitive to an increase of (dE/dx){sub e} and those that are not. A spatial distribution of the various defects within the talent track is provided and defects that are difficult to create are the closest of the ion path. The different kind of radicals created after irradiations are studied by ESR spectroscopy. Alkyl, peroxy and polyenyl radicals are detected after SHI radiations like after electron or {gamma} irradiations. Their yield of creation is independent of (dE/dx){sub e} and their localised in the crystalline zone or/and at the interfacial zone between crystalline and amorphous one. An other kind of radicals is created only after SHI radiations that are specific of the SHI-polymer interaction. We observe that these radicals are localised on a carbon cluster, in the core of the latent track for low doses and highly sensitive at the (dE/dx){sub e} of the ion. (author)

An Investigation of the Polypeptide, Poly - L - Glutamic Acid, Using Neutron Inelastic Scattering

Energy Technology Data Exchange (ETDEWEB)

Whittemore, W. L. [Gulf General Atomic Incorporated, San Diego, CA (United States)

1968-09-15

The polypeptides are synthetic polymers of amino acids with many similarities to natural proteins. In a large number of cases, one of the conformations for both the synthetic and natural proteins is the {alpha} - helix. The simplest of the synthetic polymers with no side chains is polyglycine and the simplest of the synthetic polymers with a small side chain (methyl group) is polyalanine. Dispersion curves have been computed by Gupta for both of these polymers. Polyglutamic acid is similar to polyalanine in that the composition of the basic residue and radius of helix is the same. Polyglutamic acid has a more complicated side chain which will contribute a number of additional natural frequencies that are expected to be essentially independent of conformation. On the other hand, the dispersion curves already derived for polyalanine in the {alpha} -helix form should be correct in many specific details for polyglutamic acid. An experimental study has been undertaken for polyglutamic acid at room temperature using the techniques of inelastic neutron scattering. In the first measurements, 'cold' neutrons from a reactor were used to investigate the energy level structure up to Asymptotically-Equal-To 3 kT for both conformations of the polymer. In addition, the scattering of monoenergetic high-energy neutrons ( > 0.15 eV) provided- by an electron Linac was used to study energy levels above 3 kT. These latter measurements permit comparisons to be made between the calculated and measured results for a much larger range of frequencies (and hence permit a check for a larger number of dispersion curves). This extension of the experimental results to higher frequencies has made it possible to check on the earlier assumption that only the lower frequencies are altered when the conformation is changed. This assumption underlies the evaluation of changes in internal energy with conformation from only the 'cold' neutron data, as is done with the present data. An experiment was

Perturbation correction for alanine dosimeters in different phantom materials in high-energy photon beams.

Science.gov (United States)

von Voigts-Rhetz, P; Anton, M; Vorwerk, H; Zink, K

2016-02-07

In modern radiotherapy the verification of complex treatments plans is often performed in inhomogeneous or even anthropomorphic phantoms. For dose verification small detectors are necessary and therefore alanine detectors are most suitable. Though the response of alanine for a wide range of clinical photon energies in water is well know, the knowledge about the influence of the surrounding phantom material on the response of alanine is sparse. Therefore we investigated the influence of twenty different surrounding/phantom materials for alanine dosimeters in clinical photon fields via Monte Carlo simulations. The relative electron density of the used materials was in the range [Formula: see text] up to 1.69, covering almost all materials appearing in inhomogeneous or anthropomorphic phantoms used in radiotherapy. The investigations were performed for three different clinical photon spectra ranging from 6 to 25 MV-X and Co-60 and as a result a perturbation correction [Formula: see text] depending on the environmental material was established. The Monte Carlo simulation show, that there is only a small dependence of [Formula: see text] on the phantom material and the photon energy, which is below  ±0.6%. The results confirm the good suitability of alanine detectors for in-vivo dosimetry.

Alanine racemase is essential for the growth and interspecies competitiveness of Streptococcus mutans.

Science.gov (United States)

Wei, Yuan; Qiu, Wei; Zhou, Xue-Dong; Zheng, Xin; Zhang, Ke-Ke; Wang, Shi-Da; Li, Yu-Qing; Cheng, Lei; Li, Ji-Yao; Xu, Xin; Li, Ming-Yun

2016-12-16

D-alanine (D-Ala) is an essential amino acid that has a key role in bacterial cell wall synthesis. Alanine racemase (Alr) is a unique enzyme that interconverts L-alanine and D-alanine in most bacteria, making this enzyme a potential target for antimicrobial drug development. Streptococcus mutans is a major causative factor of dental caries. The factors involved in the survival, virulence and interspecies interactions of S. mutans could be exploited as potential targets for caries control. The current study aimed to investigate the physiological role of Alr in S. mutans. We constructed alr mutant strain of S. mutans and evaluated its phenotypic traits and interspecies competitiveness compared with the wild-type strain. We found that alr deletion was lethal to S. mutans. A minimal supplement of D-Ala (150 μg·mL -1 ) was required for the optimal growth of the alr mutant. The depletion of D-alanine in the growth medium resulted in cell wall perforation and cell lysis in the alr mutant strain. We also determined the compromised competitiveness of the alr mutant strain relative to the wild-type S. mutans against other oral streptococci (S. sanguinis or S. gordonii), demonstrated using either conditioned medium assays or dual-species fluorescent in situ hybridization analysis. Given the importance and necessity of alr to the growth and competitiveness of S. mutans, Alr may represent a promising target to modulate the cariogenicity of oral biofilms and to benefit the management of dental caries.

Thermally Induced Alpha-Helix to Beta-Sheet Transition in Regenerated Silk Fibers and Films

Energy Technology Data Exchange (ETDEWEB)

Drummy,L.; Phillips, D.; Stone, M.; Farmer, B.; Naik, R.

2005-01-01

The structure of thin films cast from regenerated solutions of Bombyx mori cocoon silk in hexafluoroisopropyl alcohol (HFIP) was studied by synchrotron X-ray diffraction during heating. A solid-state conformational transition from an alpha-helical structure to the well-known beta-sheet silk II structure occurred at a temperature of approximately 140 degrees C. The transition appeared to be homogeneous, as both phases do not coexist within the resolution of the current study. Modulated differential scanning calorimetry (DSC) of the films showed an endothermic melting peak followed by an exothermic crystallization peak, both occurring near 140 degrees C. Oriented fibers were also produced that displayed this helical molecular conformation. Subsequent heating above the structural transition temperature produced oriented beta-sheet fibers very similar in structure to B. mori cocoon fibers. Heat treatment of silk films at temperatures well below their degradation temperature offers a controllable route to materials with well-defined structures and mechanical behavior.

Adsorption characteristics of 14C-labeled alanine, aspartic acid and adenosine triphosphate by metal-chelating resins

International Nuclear Information System (INIS)

Ishiyama, Toshio; Matsunami, Tadao; Shibata, Setsuko; Honda, Yoshihide.

1987-01-01

(1) Adsorption properties of 14 C-alanine, 14 C-ATP (adenosine triphosphate) and 14 C-aspartic acid on the metal-chelating resins were determined and found that the Cu(II)-Chelex 100 and Fe(III)-Unicellex UR10, Fe(III)-Chelex 100 chelating resins were highly effective for the adsorption of 14 C-alanine and 14 C-ATP, respectively. (2) Desorption rate of 14 C-ATP from the Fe(III)-Unicellex UR10 and Fe(III)-Chelex 100 resins was somewhat higher than the case of 14 C-alanine, probably because the coordination bonds of Cu-alanine might be stronger than those of Fe-ATP. Thus, 14 C-labeled organic compounds such as 14 C-alanine and 14 C-ATP of a low activity concentration (3.7 mBq/ml) (1 x 10 -7 μCi/ml) in aqueous solution may be measured with liquid scintillation counter after pre-concentration by use of the Fe(III)- and Cu(II)-chelating resin columns. (author)

Alanine dosimetry at NPL - the development of a mailed reference dosimetry service at radiotherapy dose levels

International Nuclear Information System (INIS)

Sharpe, P.H.G.; Sephton, J.P.

1999-01-01

In this paper we describe the work that has been carried out at National Physical Laboratory (NPL) to develop a mailed alanine reference dosimetry service for radiotherapy dose levels. The service is based on alanine/paraffin wax dosimeters produced at NPL. Using a data analysis technique based on spectrum fitting, it has been possible to achieve a precision of dose measurement better than ±0.05 Gy (1σ). A phantom set has been developed for use in high energy photon beams, which enables simultaneous irradiation of alanine dosimeters and ionisation chambers in a well defined geometry. Studies in photon beams of energies between 60 Co and 20 MeV have shown no significant energy dependence (<1%) for alanine relative to dose determination using a graphite calorimeter. Work is underway to extend the service to electron beams, and preliminary results are presented on the direct calibration of alanine in electron beams using a graphite calorimeter. (author)

Synthesis of {sup 15}N isotope labeled alanine; Sintese da alanina enriquecida com {sup 15}N

Energy Technology Data Exchange (ETDEWEB)

Oliveira, Claudineia R. de; Bendassolli, Jose Albertino; Sant' Ana, Carlos Roberto; Tagliassachi, Romulo Barbieri; Maximo, Everaldo; Prestes, Clelber Vieira [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil). Dept. de Isotopos Estaveis]. E-mail: crolivei@cena.usp.br

2005-07-01

The application of light chemical elements and their stable isotopes in biological studies have been increased over the last years. The use of {sup 15}N labeled amino acids is an important tool for elucidation of peptides structures. This paper describe a method for the synthesis of {sup 15}N isotope labeled alanine at lower costs than international ones, as well as the details of the recovery system of the nitrogen residues. In the present work an amination of {alpha}-haloacids, with the bromopropionic carboxylic acid and labeled aqua ammonia ({sup 15}NH{sub 3} aq) was carried out. In order to avoid eventually losses of {sup 15}NH{sub 3}, special cares were adopted, since the production cost is high. Although the acquisition cost of the {sup 13}N (radioactive) labeled compounds is lower, the obtained stable tracer will allow the accomplishment of important studies of the nitrogen cycling in living things, less occupational and environment hazards, and the time limitation problems in field studies. The tests took place in triplicates with NH{sub 3} (aq) being employed. With the establishment of the system for {sup 15}NH{sub 3} recovery, an average of 94 % of the ammonia employed in the synthesis process was recovered. The purity of the amino acid was state determined by TLC (Thin Layer Chromatography) and HPLC (High-Performance Liquid Chromatography) with a fluorescence detector. The Rf and the retention time of the synthesized sample were similar the sigma standard. Finally, regarding the established conditions, it was possible to obtain the alanine with a production cost about 40 % lower than the international price. (author)

Effects of high-salinity seawater acclimation on the levels of D-alanine in the muscle and hepatopancreas of kuruma prawn, Marsupenaeus japonicus.

Science.gov (United States)

Yoshikawa, Naoko; Yokoyama, Masahumi

2015-12-10

Changes in D- and L-alanine contents were determined in the muscle and hepatopancreas of kuruma prawn Marsupenaeus japonicus, during acclimation from seawater containing 100% salinity to artificial seawater containing 150% salinity. In the hepatopancreas, contents of both amino acids increased by approximately threefold. The activity of alanine racemase, which catalyzes the interconversion of D- and L-alanine, also increased in the high-salinity seawater. In addition, the expression of the gene encoding alanine racemase increased in the hepatopancreas with an increase in the alanine racemase activity. These data indicate that the biosynthesis of D- and L-alanine is controlled by the gene expression level of alanine racemase, and D-alanine in the hepatopancreas functions as a major osmolyte for isosmotic regulation. In contrast, the content of D-alanine and alanine racemase activity did not change in the muscle during hyper-osmotic acclimation. Therefore, we suggest that D-alanine, which exists in the several tissues of M. japonicus, is considered to be utilized in some different physiological phenomena in different tissues. Copyright © 2015 Elsevier B.V. All rights reserved.

Photochemical redox reactions of copper(II)-alanine complexes in aqueous solutions.

Science.gov (United States)

Lin, Chen-Jui; Hsu, Chao-Sheng; Wang, Po-Yen; Lin, Yi-Liang; Lo, Yu-Shiu; Wu, Chien-Hou

2014-05-19

The photochemical redox reactions of Cu(II)/alanine complexes have been studied in deaerated solutions over an extensive range of pH, Cu(II) concentration, and alanine concentration. Under irradiation, the ligand-to-metal charge transfer results in the reduction of Cu(II) to Cu(I) and the concomitant oxidation of alanine, which produces ammonia and acetaldehyde. Molar absorptivities and quantum yields of photoproducts for Cu(II)/alanine complexes at 313 nm are characterized mainly with the equilibrium Cu(II) speciation where the presence of simultaneously existing Cu(II) species is taken into account. By applying regression analysis, individual Cu(I) quantum yields are determined to be 0.094 ± 0.014 for the 1:1 complex (CuL) and 0.064 ± 0.012 for the 1:2 complex (CuL2). Individual quantum yields of ammonia are 0.055 ± 0.007 for CuL and 0.036 ± 0.005 for CuL2. Individual quantum yields of acetaldehyde are 0.030 ± 0.007 for CuL and 0.024 ± 0.007 for CuL2. CuL always has larger quantum yields than CuL2, which can be attributed to the Cu(II) stabilizing effect of the second ligand. For both CuL and CuL2, the individual quantum yields of Cu(I), ammonia, and acetaldehyde are in the ratio of 1.8:1:0.7. A reaction mechanism for the formation of the observed photoproducts is proposed.

Self-assembled RNA-triple-helix hydrogel scaffold for microRNA modulation in the tumour microenvironment

Science.gov (United States)

Conde, João; Oliva, Nuria; Atilano, Mariana; Song, Hyun Seok; Artzi, Natalie

2016-03-01

The therapeutic potential of miRNA (miR) in cancer is limited by the lack of efficient delivery vehicles. Here, we show that a self-assembled dual-colour RNA-triple-helix structure comprising two miRNAs--a miR mimic (tumour suppressor miRNA) and an antagomiR (oncomiR inhibitor)--provides outstanding capability to synergistically abrogate tumours. Conjugation of RNA triple helices to dendrimers allows the formation of stable triplex nanoparticles, which form an RNA-triple-helix adhesive scaffold upon interaction with dextran aldehyde, the latter able to chemically interact and adhere to natural tissue amines in the tumour. We also show that the self-assembled RNA-triple-helix conjugates remain functional in vitro and in vivo, and that they lead to nearly 90% levels of tumour shrinkage two weeks post-gel implantation in a triple-negative breast cancer mouse model. Our findings suggest that the RNA-triple-helix hydrogels can be used as an efficient anticancer platform to locally modulate the expression of endogenous miRs in cancer.

Strong contributions from vertical triads to helix-partner preferences in parallel coiled coils.

Science.gov (United States)

Steinkruger, Jay D; Bartlett, Gail J; Woolfson, Derek N; Gellman, Samuel H

2012-09-26

Pairing preferences in heterodimeric coiled coils are determined by complementarities among side chains that pack against one another at the helix-helix interface. However, relationships between dimer stability and interfacial residue identity are not fully understood. In the context of the "knobs-into-holes" (KIH) packing pattern, one can identify two classes of interactions between side chains from different helices: "lateral", in which a line connecting the adjacent side chains is perpendicular to the helix axes, and "vertical", in which the connecting line is parallel to the helix axes. We have previously analyzed vertical interactions in antiparallel coiled coils and found that one type of triad constellation (a'-a-a') exerts a strong effect on pairing preferences, while the other type of triad (d'-d-d') has relatively little impact on pairing tendencies. Here, we ask whether vertical interactions (d'-a-d') influence pairing in parallel coiled-coil dimers. Our results indicate that vertical interactions can exert a substantial impact on pairing specificity, and that the influence of the d'-a-d' triad depends on the lateral a' contact within the local KIH motif. Structure-informed bioinformatic analyses of protein sequences reveal trends consistent with the thermodynamic data derived from our experimental model system in suggesting that heterotriads involving Leu and Ile are preferred over homotriads involving Leu and Ile.

Reinforcement of poly (vinyl alcohol films with alpha-chitin nanowhiskers

Directory of Open Access Journals (Sweden)

Hugo Lisboa

2018-03-01

Full Text Available Abstract Composites Films were produced using Poly (Vinyl Alcohol as the soft material and reinforced with Chitin Nanowhiskers(NWCH as the rigid material. The present work studies the reinforcing mechanisms of NWCH in PVA films, made through a solvent casting technique and characterized for their calorimetric, swelling and mechanical properties. DSC tests revealed a sharp increase of 45 °C in glass transition temperatures with only 1.5% NWCH, while melting temperature had a small increases suggesting an anti-plasticizing effect. Swelling tests revealed decreasing hygoscopy when NWCH volume fraction increases. Estimates for elastic tensile modulus using a model that predicts the formation of a percolating network were not consistent with the experimental data of tensile tests suggesting that contrary to the reinforcement with cellulose nanowhiskers the percolating network is not primarily responsible for the reinforcement of the films. By adjusting the Halpin-Tsai equations, modified by Nielsen it was found that the mechanical properties were mainly influenced by the packing of the NWCH.

What can triple helix frameworks offer to the analysis of eco-innovation dynamics? Theoretical and methodological considerations

DEFF Research Database (Denmark)

Yang, Yan; Holgaard, Jette Egelund; Remmen, Arne

2012-01-01

stakeholder groups are interacting in this connection. Taking the triple helix as the theoretical departure point, this paper discusses the opportunities offered by these triple helix frameworks for analyzing eco-innovation dynamics from both theoretical and practical perspectives. It adds to the debate about......Bringing environmental concerns into focus of innovation processes will in several cases also expand the numbers of actors involved. Eco-innovation and triple helix are often frameworks applied to analyse how environmental concerns are integrated in the innovation processes and how different...