Engineering membrane scaffolds with both physical and biomolecular signaling.

PubMed

Tejeda-Montes, Esther; Smith, Katherine H; Poch, Marta; López-Bosque, María Jesús; Martín, Laura; Alonso, Matilde; Engel, Elisabeth; Mata, Alvaro

2012-03-01

We report on the combination of a top-down and bottom-up approach to develop thin bioactive membrane scaffolds based on functional elastin-like polymers (ELPs). Our strategy combines ELP cross-linking and assembly, and a variety of standard and novel micro/nanofabrication techniques to create self-supporting membranes down to ∼500 nm thick that incorporate both physical and biomolecular signals, which can be easily tailored for a specific application. In this study we used an ELP that included the cell-binding motif arginine-glycine-aspartic acid-serine (RGDS). Furthermore, fabrication processes were developed to create membranes that exhibited topographical patterns with features down to 200 nm in lateral dimensions and up to 10 μm in height on either one or both sides, uniform and well-defined pores, or multiple ELP layers. A variety of processing parameters were tested in order to optimize membrane fabrication, including ELP and cross-linker concentration, temperature, reaction time and ambient humidity. Membrane micro/nanopatterning, swelling and stiffness were characterized by atomic force microscopy, nanoindentation tests and scanning electron microscopy. Upon immersion in phosphate-buffered saline and an increase in temperature from 25 to 40°C, membranes exhibited a significant increase in surface stiffness, with the reduced Young's modulus increasing with temperature. Finally, rat mesenchymal stem cells were cultured on thin RGDS-containing membranes, which allowed cell adhesion, qualitatively enhanced spreading compared to membranes without RGDS epitopes and permitted proliferation. Furthermore, cell morphology was drastically affected by topographical patterns on the surface of the membranes. Copyright © 2011 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Immune-tolerant elastin-like polypeptides (iTEPs) and their application as CTL vaccine carriers.

PubMed

Cho, S; Dong, S; Parent, K N; Chen, M

2016-01-01

Cytotoxic T lymphocyte (CTL) vaccine carriers are known to enhance the efficacy of vaccines, but a search for more effective carriers is warranted. Elastin-like polypeptides (ELPs) have been examined for many medical applications but not as CTL vaccine carriers. We aimed to create immune tolerant ELPs using a new polypeptide engineering practice and create CTL vaccine carriers using the ELPs. Four sets of novel ELPs, termed immune-tolerant elastin-like polypeptide (iTEP) were generated according to the principles dictating humoral immunogenicity of polypeptides and phase transition property of ELPs. The iTEPs were non-immunogenic in mice. Their phase transition feature was confirmed through a turbidity assay. An iTEP nanoparticle (NP) was assembled from an amphiphilic iTEP copolymer plus a CTL peptide vaccine, SIINFEKL. The NP facilitated the presentation of the vaccine by dendritic cells (DCs) and enhanced vaccine-induced CTL responses. A new ELP design and development practice was established. The non-canonical motif and the immune tolerant nature of the iTEPs broaden our insights about ELPs. ELPs, for the first time, were successfully used as carriers for CTL vaccines. It is feasible to concurrently engineer both immune-tolerant and functional peptide materials. ELPs are a promising type of CTL vaccine carriers.

Dual stimuli-sensitive dendrimers: Photothermogenic gold nanoparticle-loaded thermo-responsive elastin-mimetic dendrimers.

PubMed

Fukushima, Daichi; Sk, Ugir Hossain; Sakamoto, Yasuhiro; Nakase, Ikuhiko; Kojima, Chie

2015-08-01

Dendrimers are synthetic macromolecules with unique structures that can work as nanoplatforms for both photothermogenic gold nanoparticles (AuNPs) and thermosensitive elastin-like peptides (ELPs) with valine-proline-glycine-valine-glycine (VPGVG) repeats. In this study, photothermogenic AuNPs were loaded into thermo-responsive elastin-mimetic dendrimers (dendrimers conjugating ELPs at their periphery) to produce dual stimuli-sensitive nanoparticles. Polyamidoamine G4 dendrimers were modified with acetylated VPGVG and (VPGVG)2, and the resulting materials were named ELP1-den and ELP2-den, respectively. The AuNPs were prepared by the reduction of Au ions using a dendrimer-nanotemplated method. The AuNP-loaded elastin-mimetic dendrimers exhibited photothermal properties. ELP1-den and ELP2-den showed similar temperature-dependent changes in their conformations. Phase transitions were observed at around 55°C and 35°C for the AuNP-loaded ELP1-den and AuNP-loaded ELP2-den, respectively, but not for the corresponding PEGylated dendrimer. In contrast to the AuNP-loaded PEGylated dendrimer, AuNP-loaded ELP2-den readily associated with cells and induced efficient photocytotoxicity at 37°C. The cell association and the photocytotoxicity properties of AuNP-loaded ELP2-den could be controlled by temperature. These results therefore suggest that dual stimuli-sensitive dendrimer nanoparticles of this type could be used for photothermal therapy. Copyright © 2015 Elsevier B.V. All rights reserved.

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

PubMed Central

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

2012-01-01

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

Exon Specific U1 snRNAs improve ELP1 exon 20 definition and rescue ELP1 protein expression in a Familial Dysautonomia mouse model.

PubMed

Donadon, Irving; Pinotti, Mirko; Rajkowska, Katarzyna; Pianigiani, Giulia; Barbon, Elena; Morini, Elisabetta; Motaln, Helena; Rogelj, Boris; Mingozzi, Federico; Slaugenhaupt, Susan A; Pagani, Franco

2018-04-25

Familial dysautonomia (FD) is a rare genetic disease with no treatment, caused by an intronic point mutation (c.2204 + 6T>C) that negatively affects the definition of exon 20 in the Elongator complex protein 1 gene (ELP1 also known as IKBKAP). This substitution modifies the 5' splice site and, in combination with regulatory splicing factors, induces different levels of exon 20 skipping, in various tissues. Here, we evaluated the therapeutic potential of a novel class of U1 snRNA molecules, Exon-Specific U1s (ExSpeU1s), in correcting ELP1 exon 20 recognition. Lentivirus-mediated expression of ELP1-ExSpeU1 in FD fibroblasts improved ELP1 splicing and protein levels. We next focused on a transgenic mouse model that recapitulates the same tissue-specific mis-splicing seen in FD patients. Intraperitoneal delivery of ELP1-ExSpeU1s-adeno-associated virus particles successfully increased the production of full-length human ELP1 transcript and protein. This splice-switching class of molecules is the first to specifically correct the ELP1 exon 20 splicing defect. Our data provide proof of principle of ExSpeU1s-adeno-associated virus particles as a novel therapeutic strategy for FD.

Proliferative activity of elastin-like-peptides depends on charge and phase transition.

PubMed

Yuan, Yuan; Koria, Piyush

2016-03-01

Elastin-like-peptides (ELPs) are stimulus-responsive protein-based polymers and are attractive biomaterials due to their biocompatibility and unique properties. This study shows that in addition to their physical properties, ELPs have biological activities that are conducive to tissue regeneration. Specifically, we found that ELPs induce fibroblast proliferation via cell surface heparan sulfate proteoglycans (HSPG). Furthermore, our data suggests that ELP based materials with differential proliferative potential can be designed by controlling the interaction of ELPs with HSPGs by incorporating either hydrophobic or positively charged residues within the ELP sequence. Fibroblast proliferation is important for granulation tissue formation which is important in chronic wounds as well as in healing of other tissues. The customizable biological activity of ELPs coupled with their unique physical properties will enable us to design novel, sustainable and cost effective therapies for different tissue regeneration applications. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 697-706, 2016. © 2015 Wiley Periodicals, Inc.

Selective Tuning of Elastin-like Polypeptide Properties via Methionine Oxidation.

PubMed

Petitdemange, Rosine; Garanger, Elisabeth; Bataille, Laure; Dieryck, Wilfrid; Bathany, Katell; Garbay, Bertrand; Deming, Timothy J; Lecommandoux, Sébastien

2017-02-13

We have designed and prepared a recombinant elastin-like polypeptide (ELP) containing precisely positioned methionine residues, and performed the selective and complete oxidation of its methionine thioether groups to both sulfoxide and sulfone derivatives. Since these oxidation reactions substantially increase methionine residue polarity, they were found to be a useful means to precisely adjust the temperature responsive behavior of ELPs in aqueous solutions. In particular, lower critical solution temperatures were found to be elevated in oxidized sample solutions, but were not eliminated. These transition temperatures were found to be further tunable by the use of solvents containing different Hofmeister salts. Overall, the ability to selectively and fully oxidize methionine residues in ELPs proved to be a convenient postmodification strategy for tuning their transition temperatures in aqueous media.

The Arabidopsis ELP3/ELO3 and ELP4/ELO1 genes enhance disease resistance in Fragaria vesca L.

PubMed

Silva, Katchen Julliany P; Brunings, Asha M; Pereira, Juliana A; Peres, Natalia A; Folta, Kevin M; Mou, Zhonglin

2017-12-01

Plant immune response is associated with a large-scale transcriptional reprogramming, which is regulated by numerous transcription regulators such as the Elongator complex. Elongator is a multitasking protein complex involved in diverse cellular processes, including histone modification, DNA methylation, and tRNA modification. In recent years, Elongator is emerging as a key regulator of plant immune responses. However, characterization of Elongator's function in plant immunity has been conducted only in the model plant Arabidopsis thaliana. It is thus unclear whether Elongator's role in plant immunity is conserved in higher plants. The objective of this study is to characterize transgenic woodland strawberry (Fragaria vesca L.) overexpressing the Arabidopsis Elongator (AtELP) genes, AtELP3 and AtELP4, and to determine whether F. vesca carries a functional Elongator complex. Transgenic F. vesca and Arabidopsis plants were produced via Agrobacterium-mediated genetic transformation and characterized by morphology, PCR, real-time quantitative PCR, and disease resistance test. The Student's t test was used to analyze the data. Overexpression of AtELP3 and AtELP4 in F. vesca impacts plant growth and development and confers enhanced resistance to anthracnose crown rot, powdery mildew, and angular leaf spot, which are caused by the hemibiotrophic fungal pathogen Colletotrichum gloeosporioides, the obligate biotrophic fungal pathogen Podosphaera aphanis, and the hemibiotrophic bacterial pathogen Xanthomonas fragariae, respectively. Moreover, the F. vesca genome encodes all six Elongator subunits by single-copy genes with the exception of FvELP4, which is encoded by two homologous genes, FvELP4-1 and FvELP4-2. We show that FvELP4-1 complemented the Arabidopsis Atelp4/elo1-1 mutant, indicating that FvELP4 is biologically functional. This is the first report on overexpression of Elongator genes in plants. Our results indicate that the function of Elongator in plant immunity is most likely conserved in F. vesca and suggest that Elongator genes may hold potential for helping mitigate disease severity and reduce the use of fungicides in strawberry industry.

Arabidopsis Elongator subunit 2 positively contributes to resistance to the necrotrophic fungal pathogens Botrytis cinerea and Alternaria brassicicola.

PubMed

Wang, Chenggang; Ding, Yezhang; Yao, Jin; Zhang, Yanping; Sun, Yijun; Colee, James; Mou, Zhonglin

2015-09-01

The evolutionarily conserved Elongator complex functions in diverse biological processes including salicylic acid-mediated immune response. However, how Elongator functions in jasmonic acid (JA)/ethylene (ET)-mediated defense is unknown. Here, we show that Elongator is required for full induction of the JA/ET defense pathway marker gene PLANT DEFENSIN1.2 (PDF1.2) and for resistance to the necrotrophic fungal pathogens Botrytis cinerea and Alternaria brassicicola. A loss-of-function mutation in the Arabidopsis Elongator subunit 2 (ELP2) alters B. cinerea-induced transcriptome reprogramming. Interestingly, in elp2, expression of WRKY33, OCTADECANOID-RESPONSIVE ARABIDOPSIS AP2/ERF59 (ORA59), and PDF1.2 is inhibited, whereas transcription of MYC2 and its target genes is enhanced. However, overexpression of WRKY33 or ORA59 and mutation of MYC2 fail to restore PDF1.2 expression and B. cinerea resistance in elp2, suggesting that ELP2 is required for induction of not only WRKY33 and ORA59 but also PDF1.2. Moreover, elp2 is as susceptible as coronatine-insensitive1 (coi1) and ethylene-insensitive2 (ein2) to B. cinerea, indicating that ELP2 is an important player in B. cinerea resistance. Further analysis of the lesion sizes on the double mutants elp2 coi1 and elp2 ein2 and the corresponding single mutants revealed that the function of ELP2 overlaps with COI1 and is additive to EIN2 for B. cinerea resistance. Finally, basal histone acetylation levels in the coding regions of WRKY33, ORA59, and PDF1.2 are reduced in elp2 and a functional ELP2-GFP fusion protein binds to the chromatin of these genes, suggesting that constitutive ELP2-mediated histone acetylation may be required for full activation of the WRKY33/ORA59/PDF1.2 transcriptional cascade. © 2015 The Authors The Plant Journal © 2015 John Wiley & Sons Ltd.

Elongator Plays a Positive Role in Exogenous NAD-Induced Defense Responses in Arabidopsis.

PubMed

An, Chuanfu; Ding, Yezhang; Zhang, Xudong; Wang, Chenggang; Mou, Zhonglin

2016-05-01

Extracellular NAD is emerging as an important signal molecule in animal cells, but its role in plants has not been well-established. Although it has been shown that exogenous NAD(+) activates defense responses in Arabidopsis, components in the exogenous NAD(+)-activated defense pathway remain to be fully discovered. In a genetic screen for mutants insensitive to exogenous NAD(+) (ien), we isolated a mutant named ien2. Map-based cloning revealed that IEN2 encodes ELONGATA3 (ELO3)/AtELP3, a subunit of the Arabidopsis Elongator complex, which functions in multiple biological processes, including histone modification, DNA (de)methylation, and transfer RNA modification. Mutations in the ELO3/AtELP3 gene compromise exogenous NAD(+)-induced expression of pathogenesis-related (PR) genes and resistance to the bacterial pathogen Pseudomonas syringae pv. maculicola ES4326, and transgenic expression of the coding region of ELO3/AtELP3 in elo3/Atelp3 restores NAD(+) responsiveness to the mutant plants, demonstrating that ELO3/AtELP3 is required for exogenous NAD(+)-induced defense responses. Furthermore, mutations in genes encoding the other five Arabidopsis Elongator subunits (ELO2/AtELP1, AtELP2, ELO1/AtELP4, AtELP5, and AtELP6) also compromise exogenous NAD(+)-induced PR gene expression and resistance to P. syringae pv. maculicola ES4326. These results indicate that the Elongator complex functions as a whole in exogenous NAD(+)-activated defense signaling in Arabidopsis.

Mammary cell-activating factor regulates the hormone-independent transcription of the early lactation protein (ELP) gene in a marsupial.

PubMed

Pharo, Elizabeth A; Renfree, Marilyn B; Cane, Kylie N

2016-11-15

The regulation of the tammar wallaby (Macropus eugenii) early lactation protein (ELP) gene is complex. ELP is responsive to the lactogenic hormones; insulin (I), hydrocortisone (HC) and prolactin (PRL) in mammary gland explants but could not be induced with lactogenic hormones in tammar primary mammary gland cells, nor in KIM-2 conditionally immortalised murine mammary epithelial cells. Similarly, ELP promoter constructs transiently-transfected into human embryonic kidney (HEK293T) cells constitutively expressing the prolactin receptor (PRLR) and Signal Transducer and Activator of Transcription (STAT)5A were unresponsive to prolactin, unlike the rat and mouse β-casein (CSN2) promoter constructs. Identification of the minimal promoter required for the hormone-independent transcription of tammar ELP in HEK293Ts and comparative analysis of the proximal promoters of marsupial ELP and the orthologous eutherian colostrum trypsin inhibitor (CTI) gene suggests that mammary cell-activating factor (MAF), an E26 transformation-specific (ETS) factor, may bind to an AGGAAG motif and activate tammar ELP. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Imparting large macroscopic changes with small changes in polypeptide composition

NASA Astrophysics Data System (ADS)

Sing, Michelle; McKinley, Gareth; Olsen, Bradley

Block copolymers composed of polypeptides provide an excellent platform for exploring the underlying physics surrounding macroscopic associative network behavior. Previous work in our group has elucidated a difference in the mechanical properties of two nearly identical elastin-like polypeptide (ELP) endblocks. In poly(ELP)s, this substitution is known to result in tighter beta turns. These beta turns exhibit slower responses to changes in temperature within the material. Under shear, the modulus for the alanine-containing ELP triblock is almost three times higher than the glycine-containing ELP. Additionally, preliminary tensile tests show higher stress and strain at break for the alanine ELP triblock. We are able to explain the reasons for this behavior using a variety of spectroscopic and analytical techniques. Small angle neutron and x-ray scattering indicate differences in ordering between the alanine and glycine containing ELP materials both in shear and in stagnant flow.

Improvement of physical properties of calcium phosphate cement by elastin-like polypeptide supplementation.

PubMed

Jang, Ji-Hyun; Shin, Sumi; Kim, Hyun-Jung; Jeong, Jinyoung; Jin, Hyo-Eon; Desai, Malav S; Lee, Seung-Wuk; Kim, Sun-Young

2018-03-26

Calcium phosphate cements (CPCs) are synthetic bioactive cements widely used as hard tissue substitutes. Critical limitations of use include their poor mechanical properties and poor anti-washout behaviour. To address those limitations, we combined CPC with genetically engineered elastin-like polypeptides (ELPs). We investigated the effect of the ELPs on the physical properties and biocompatibility of CPC by testing ELP/CPC composites with various liquid/powder ratios. Our results show that the addition of ELPs improved the mechanical properties of the CPC, including the microhardness, compressive strength, and washout resistance. The biocompatibility of ELP/CPC composites was also comparable to that of the CPC alone. However, supplementing CPC with ELPs functionalized with octaglutamate as a hydroxyapatite binding peptide increased the setting time of the cement. With further design and modification of our biomolecules and composites, our research will lead to products with diverse applications in biology and medicine.

Elastin-like polypeptide switches: A design strategy to detect multimeric proteins.

PubMed

Dhandhukia, Jugal P; Brill, Dab A; Kouhi, Aida; Pastuszka, Martha K; MacKay, J Andrew

2017-09-01

Elastin-Like Polypeptides (ELPs) reversibly phase separate in response to changes in temperature, pressure, concentration, pH, and ionic species. While powerful triggers, biological microenvironments present a multitude of more specific biological cues, such as antibodies, cytokines, and cell-surface receptors. To develop better biosensors and bioresponsive drug carriers, rational strategies are required to sense and respond to these target proteins. We recently reported that noncovalent association of two ELP fusion proteins to a "chemical inducer of dimerization" small molecule (1.5 kDa) induces phase separation at physiological temperatures. Having detected a small molecule, here we present the first evidence that ELP multimerization can also detect a much larger (60 kDa) protein target. To demonstrate this strategy, ELPs were biotinylated at their amino terminus and mixed with tetrameric streptavidin. At a stoichiometric ratio of [4:1], two to three biotin-ELPs associate with streptavidin into multimeric complexes with an apparent K d of 5 nM. The increased ELP density around a streptavidin core strongly promotes isothermal phase separation, which was tuned to occur at physiological temperature. This phase separation reverses upon saturation with excess streptavidin, which only favors [1:1] complexes. Together, these findings suggest that ELP association with multimeric biomolecules is a viable strategy to deliberately engineer ELPs that respond to multimeric protein substrates. © 2017 The Protein Society.

SU-F-T-10: Validation of ELP Dosimetry Using PRESAGE Dosimeter: Feasibility Test and Practical Considerations

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

Lambson, K; Lafata, K; Miles, D

Purpose: To validate the use of a PRESAGE dosimeter as a method to quantitatively measure dose distributions of injectable brachytherapy based on elastin-like polypeptide (ELP) nanoparticles. PRESAGE is a solid, transparent polyurethane-based dosimeter whose dose is proportional to a change in optical density, making it useful for visualizing the dose from a radionuclide-tagged-ELP injection. Methods: A PRESAGE dosimeter was designed to simulate an ELP injection. To calibrate, cuvette samples from the batch of PRESAGE were exposed to varying levels of radiation from 0–35.9Gy applied via a linear accelerator, then placed into a spectrophotometer to obtain the optical density change asmore » a function of dose. A pre-optical-CT scan was acquired of the phantom to obtain a baseline tomographic optical density. A 1cc saline solution of I-125 tagged-ELP with and activity concentration of 1mCi/cc was injected into the phantom and left for five days. After five days, the ELP was removed and the cavity cleaned of all remaining radioactive material. Post tomographic optical images were acquired to obtain a differential optical density dataset. Results: Initial results after the 5-day exposure revealed an opaque white film that resembled the volume of the ELP solution injected into the phantom. We think this is possibly due to the saline solution diffusing into the PRESAGE and causing a change in the index of refraction at this shallow depth. Therefore, initially the optical scanner yielded inconclusive results. After several more days, the saline seemed to have evaporated out of the injection site and the ELP dose distribution was visible via color change in the dosimeter. Conclusion: We have created the first experimental design to measure the dose distribution of I-125-tagged-ELP. The PRESAGE formulation proves to be a feasible option for such measurements. Future experimental measurements need to be obtained to further characterize ELP dosimetry.« less

Inhibition of ovarian cancer cell proliferation by a cell cycle inhibitory peptide fused to a thermally responsive polypeptide carrier.

PubMed

Massodi, Iqbal; Moktan, Shama; Rawat, Aruna; Bidwell, Gene L; Raucher, Drazen

2010-01-15

Current treatment of solid tumors is limited by normal tissue tolerance, resulting in a narrow therapeutic index. To increase drug specificity and efficacy and to reduce toxicity in normal tissues, we have developed a polypeptide carrier for a cell cycle inhibitory peptide, which has the potential to be thermally targeted to the tumor site. The design of this polypeptide is based on elastin-like polypeptide (ELP). The coding sequence of ELP was modified by the addition of the cell penetrating peptide Bac-7 at the N-terminus and a 23 amino acid peptide derived from p21 at the C-terminus (Bac-ELP1-p21). Bac-ELP1-p21 is soluble in aqueous solutions below physiological temperature (37 degrees C) but aggregates when the temperature is raised above 39 degrees C, making it a promising thermally responsive therapeutic carrier that may be actively targeted to solid tumors by application of focused hyperthermia. While Bac-ELP1-p21 at 37 degrees C did not have any effect on SKOV-3 cell proliferation, the use of hyperthermia increased the antiproliferative effect of Bac-ELP1-p21 compared with a thermally unresponsive control polypeptide. Bac-ELP1-p21 displayed both a cytoplasmic and nuclear distribution in the SKOV-3 cells, with nuclear-localized polypeptide enriched in the heated cells, as revealed by confocal microscopy. Using Western blotting, we show that Bac-ELP1-p21 caused a decrease in Rb phosphorylation levels in cells treated at 42 degrees C. The polypeptide also induced caspase activation, PARP cleavage, and cell cycle arrest in S-phase and G2/M-phase. These studies indicate that ELP is a promising macromolecular carrier for the delivery of cell cycle inhibitory peptides to solid tumors.

Hybrid elastin-like polypeptide-polyethylene glycol (ELP-PEG) hydrogels with improved transparency and independent control of matrix mechanics and cell ligand density.

PubMed

Wang, Huiyuan; Cai, Lei; Paul, Alexandra; Enejder, Annika; Heilshorn, Sarah C

2014-09-08

Hydrogels have been developed as extracellular matrix (ECM) mimics both for therapeutic applications and basic biological studies. In particular, elastin-like polypeptide (ELP) hydrogels, which can be tuned to mimic several biochemical and physical characteristics of native ECM, have been constructed to encapsulate various types of cells to create in vitro mimics of in vivo tissues. However, ELP hydrogels become opaque at body temperature because of ELP's lower critical solution temperature behavior. This opacity obstructs light-based observation of the morphology and behavior of encapsulated cells. In order to improve the transparency of ELP hydrogels for better imaging, we have designed a hybrid ELP-polyethylene glycol (PEG) hydrogel system that rapidly cross-links with tris(hydroxymethyl) phosphine (THP) in aqueous solution via Mannich-type condensation. As expected, addition of the hydrophilic PEG component significantly improves the light transmittance. Coherent anti-Stokes Raman scattering (CARS) microscopy reveals that the hybrid ELP-PEG hydrogels have smaller hydrophobic ELP aggregates at 37 °C. Importantly, this hydrogel platform enables independent tuning of adhesion ligand density and matrix stiffness, which is desirable for studies of cell-matrix interactions. Human fibroblasts encapsulated in these hydrogels show high viability (>98%) after 7 days of culture. High-resolution confocal microscopy of encapsulated fibroblasts reveals that the cells adopt a more spread morphology in response to higher RGD ligand concentrations and softer gel mechanics.

Cell penetrating peptides fused to a thermally targeted biopolymer drug carrier improve the delivery and antitumor efficacy of an acid-sensitive doxorubicin derivative.

PubMed

Walker, Leslie; Perkins, Eddie; Kratz, Felix; Raucher, Drazen

2012-10-15

Elastin-like polypeptide (ELP) is a macromolecular carrier with thermally responsive properties that can passively accumulate in solid tumors and additionally aggregate in tumor tissue when exposed to hyperthermia. In this study, ELP was conjugated to the anticancer drug doxorubicin (DOXO) and three different cell penetrating peptides (CPP) in order to inhibit tumor growth in mice compared to free doxorubicin. Fluorescence microscopy studies in MCF-7 breast carcinoma cells demonstrated that the three different CPP-ELP-DOXO conjugates delivered doxorubicin to the cell nucleus. All CPP-ELP-DOXO conjugates showed cytotoxicity with IC(50) values in the range of 12-30 μM at 42 °C, but the ELP carrier with SynB1 as the cell penetrating peptide had the lowest intrinsic cytotoxicity. Therefore, the antitumor efficacy of SynB1-ELP-DOXO was compared to doxorubicin under hyperthermic conditions. C57BL/6 female mice bearing syngeneic E0771 murine breast tumors were treated with either free doxorubicin or the SynB1-ELP-DOXO conjugate with or without focused hyperthermia on the tumor. Under hyperthermic conditions, tumor inhibition with SynB1-ELP-DOXO was 2-fold higher than under therapy with free doxorubicin at the equivalent dose, and is thus a promising lead candidate for optimizing thermally responsive drug polymer conjugates. Copyright © 2012 Elsevier B.V. All rights reserved.

Elp3 and Dph3 of Schizosaccharomyces pombe mediate cellular stress responses through tRNALysUUU modifications.

PubMed

Villahermosa, Desirée; Fleck, Oliver

2017-08-03

Efficient protein synthesis in eukaryotes requires diphthamide modification of translation elongation factor eEF2 and wobble uridine modifications of tRNAs. In higher eukaryotes, these processes are important for preventing neurological and developmental defects and cancer. In this study, we used Schizosaccharomyces pombe as a model to analyse mutants defective in eEF2 modification (dph1Δ), in tRNA modifications (elp3Δ), or both (dph3Δ) for sensitivity to cytotoxic agents and thermal stress. The dph3Δ and elp3Δ mutants were sensitive to a range of drugs and had growth defects at low temperature. dph3Δ was epistatic with dph1Δ for sensitivity to hydroxyurea and methyl methanesulfonate, and with elp3Δ for methyl methanesulfonate and growth at 16 °C. The dph1Δ and dph3Δ deletions rescued growth defects of elp3Δ in response to thiabendazole and at 37 °C. Elevated tRNA Lys UUU levels suppressed the elp3Δ phenotypes and some of the dph3Δ phenotypes, indicating that lack of tRNA Lys UUU modifications were responsible. Furthermore, we found positive genetic interactions of elp3Δ and dph3Δ with sty1Δ and atf1Δ, indicating that Elp3/Dph3-dependent tRNA modifications are important for efficient biosynthesis of key factors required for accurate responses to cytotoxic stress conditions.

Preparing Bilingual Teacher Candidates: A Linguistic Conundrum in a Changing Political Landscape

ERIC Educational Resources Information Center

Yturriago, Judith Kwiat; Gil-Garcia, Ana

2010-01-01

In schools across the country, many students who consistently score below their white peers on state standardized tests are English language learners (ELLs). Under No Child Left Behind (NCLB) and the reauthorized ESEA under President Obama, all 50 states are and will be required to have English language proficiency (ELP) standards and state…

Enterocolic lymphocytic phlebitis: clinicopathologic features and review of the literature.

PubMed

Ngo, Nyethane; Chang, Fuju

2007-07-01

Enterocolic lymphocytic phlebitis (ELP) is a recently described entity and is of unknown etiology and pathogenesis. It is characterized by phlebitis of the bowel wall and mesentery, without arterial involvement or evidence of systemic vasculitis. The clinical presentation of ELP is varied, but it most commonly manifests with signs of an acute abdomen. Clinical, radiologic, and endoscopic findings are often conflicting and misdiagnosis is common as venous thrombosis is not suspected. The diagnosis of ELP is obtained histologically. There is a spectrum of histologic features associated with ELP, which includes lymphocytic phlebitis, necrotizing phlebitis, granulomatous phlebitis, and myointimal hyperplasia. Other features include venous thrombi and acute ischemic changes of the intestine. Surgical resection of the affected bowel is usually curative and recurrences are rare. The clinical and histopathologic features of ELP are reviewed.

Evolution and function of eukaryotic-like proteins from sponge symbionts.

PubMed

Reynolds, David; Thomas, Torsten

2016-10-01

Sponges (Porifera) are ancient metazoans that harbour diverse microorganisms, whose symbiotic interactions are essential for the host's health and function. Although symbiosis between bacteria and sponges are ubiquitous, the molecular mechanisms that control these associations are largely unknown. Recent (meta-) genomic analyses discovered an abundance of genes encoding for eukaryotic-like proteins (ELPs) in bacterial symbionts from different sponge species. ELPs belonging to the ankyrin repeat (AR) class from a bacterial symbiont of the sponge Cymbastela concentrica were subsequently found to modulate amoebal phagocytosis. This might be a molecular mechanism, by which symbionts can control their interaction with the sponge. In this study, we investigated the evolution and function of ELPs from other classes and from symbionts found in other sponges to better understand the importance of ELPs for bacteria-eukaryote interactions. Phylogenetic analyses showed that all of the nine ELPs investigated were most closely related to proteins found either in eukaryotes or in bacteria that can live in association with eukaryotes. ELPs were then recombinantly expressed in Escherichia coli and exposed to the amoeba Acanthamoeba castellanii, which is functionally analogous to phagocytic cells in sponges. Phagocytosis assays with E. coli containing three ELP classes (AR, TPR-SEL1 and NHL) showed a significantly higher percentage of amoeba containing bacteria and average number of intracellular bacteria per amoeba when compared to negative controls. The result that various classes of ELPs found in symbionts of different sponges can modulate phagocytosis indicates that they have a broader function in mediating bacteria-sponge interactions. © 2016 John Wiley & Sons Ltd.

The effective local potential method: Implementation for molecules and relation to approximate optimized effective potential techniques

NASA Astrophysics Data System (ADS)

Izmaylov, Artur F.; Staroverov, Viktor N.; Scuseria, Gustavo E.; Davidson, Ernest R.; Stoltz, Gabriel; Cancès, Eric

2007-02-01

We have recently formulated a new approach, named the effective local potential (ELP) method, for calculating local exchange-correlation potentials for orbital-dependent functionals based on minimizing the variance of the difference between a given nonlocal potential and its desired local counterpart [V. N. Staroverov et al., J. Chem. Phys. 125, 081104 (2006)]. Here we show that under a mildly simplifying assumption of frozen molecular orbitals, the equation defining the ELP has a unique analytic solution which is identical with the expression arising in the localized Hartree-Fock (LHF) and common energy denominator approximations (CEDA) to the optimized effective potential. The ELP procedure differs from the CEDA and LHF in that it yields the target potential as an expansion in auxiliary basis functions. We report extensive calculations of atomic and molecular properties using the frozen-orbital ELP method and its iterative generalization to prove that ELP results agree with the corresponding LHF and CEDA values, as they should. Finally, we make the case for extending the iterative frozen-orbital ELP method to full orbital relaxation.

Comprehensive analysis of Arabidopsis expression level polymorphisms with simple inheritance

PubMed Central

Plantegenet, Stephanie; Weber, Johann; Goldstein, Darlene R; Zeller, Georg; Nussbaumer, Cindy; Thomas, Jérôme; Weigel, Detlef; Harshman, Keith; Hardtke, Christian S

2009-01-01

In Arabidopsis thaliana, gene expression level polymorphisms (ELPs) between natural accessions that exhibit simple, single locus inheritance are promising quantitative trait locus (QTL) candidates to explain phenotypic variability. It is assumed that such ELPs overwhelmingly represent regulatory element polymorphisms. However, comprehensive genome-wide analyses linking expression level, regulatory sequence and gene structure variation are missing, preventing definite verification of this assumption. Here, we analyzed ELPs observed between the Eil-0 and Lc-0 accessions. Compared with non-variable controls, 5′ regulatory sequence variation in the corresponding genes is indeed increased. However, ∼42% of all the ELP genes also carry major transcription unit deletions in one parent as revealed by genome tiling arrays, representing a >4-fold enrichment over controls. Within the subset of ELPs with simple inheritance, this proportion is even higher and deletions are generally more severe. Similar results were obtained from analyses of the Bay-0 and Sha accessions, using alternative technical approaches. Collectively, our results suggest that drastic structural changes are a major cause for ELPs with simple inheritance, corroborating experimentally observed indel preponderance in cloned Arabidopsis QTL. PMID:19225455

Applications of elastin-like polypeptides in drug delivery

PubMed Central

MacEwan, Sarah R; Chilkoti, Ashutosh

2014-01-01

Elastin-like polypeptides (ELPs) are biopolymers inspired by human elastin. Their lower critical solution temperature phase transition behavior and biocompatibility make them useful materials for stimulus-responsive applications in biological environments. Due to their genetically encoded design and recombinant synthesis, the sequence and size of ELPs can be exactly defined. These design parameters control the structure and function of the ELP with a precision that is unmatched by synthetic polymers. Due to these attributes, ELPs have been used extensively for drug delivery in a variety of different embodiments—as soluble macromolecular carriers, self-assembled nanoparticles, cross-linked microparticles, or thermally coacervated depots. These ELP systems have been used to deliver biologic therapeutics, radionuclides, and small molecule drugs to a variety of anatomical sites for the treatment of diseases including cancer, type 2 diabetes, osteoarthritis, and neuroinflammation. PMID:24979207

Simulation Studies of LCST-like Phase Transitions in Elastin-like Polypeptides (ELPs) and Conjugates of ELP with Rigid Macromolecules

NASA Astrophysics Data System (ADS)

Condon, Joshua; Martin, Tyler; Jayaraman, Arthi

We use atomistic (AA) and coarse-grained (CG) molecular dynamics simulations to elucidate the thermodynamic driving forces governing lower critical solution temperature (LCST)-like phase transition exhibited by elastin-like peptides (ELPs) and conjugates of ELP with other macromolecules. In the AA simulations, we study ELP oligomers in explicit water, and mark the transition as the temperature at which they undergo a change in ``hydration'' state. While AA simulations are restricted to small systems of short ELPs and do not capture the chain aggregation observed in experiments of ELPs, they guide the phenomenological CG model development by highlighting the solvent induced polymer-polymer effective interactions with changing temperature. In the CG simulations, we capture the LCST polymer aggregation by increasing polymer-polymer effective attractive interactions in an implicit solvent. We examine the impact of conjugating a block of LCST polymer to another rigid unresponsive macromolecular block on the LCST-like transition. We find that when multiple LCST polymers are conjugated to a rigid polymer block, increased crowding of the LCST polymers shifts the onset of chain aggregation to smaller effective polymer-polymer attraction compared to the free LCST polymers. These simulation results provide guidance on the design of conjugated bio-mimetic thermoresponsive materials, and shape the fundamental understanding of the impact of polymer crowding on phase behavior in thermoresponsive LCST polymer systems.

Re-Conceptualizing the ELP as a Web 2.0 Personal Language Learning Environment

ERIC Educational Resources Information Center

Haines, Kevin; van Engen, Jeroen

2013-01-01

This paper addresses the reconceptualization of the ELP as a Personal Language Learning Environment (PLLE), encouraging learners towards greater self-regulation. Such a development fits in with the pedagogical function of the ELP by scaffolding the plurilingual, lifelong learning of languages. Web 2.0 social media tools allow learners to work with…

Revisiting the Principles and Functions of the European Language Portfolio with a Focus on Tertiary Education

ERIC Educational Resources Information Center

Schärer, Rolf

2012-01-01

The European Language Portfolio (ELP) is based on principles which tend to challenge traditional learning and teaching practices and thus impact on education policy and systems. The primary owner of the ELP is the learner, and as a personal language learning companion, the ELP has three core functions: pedagogic, reporting and catalytic. For it to…

English language proficiency and physical activity among Mexican-origin women in South Texas and South Carolina.

PubMed

Salinas, Jennifer J; Hilfinger Messias, DeAnne K; Morales-Campos, Daisy; Parra-Medina, Deborah

2014-02-01

To examine the relationship between English language proficiency (ELP), physical activity, and physical activity-related psychosocial measures (i.e., exercise self-efficacy, exercise social support, perceptions of environmental supports) among Mexican-origin women in South Carolina and Texas. Adjusted robust regression and interaction modeling to evaluate baseline questionnaire data on self-reported ELP with CHAMPS leisure-time moderate-to-vigorous physical activity (MVPA), accelerometry data, Physical Activity Self-Efficacy, Physical Activity Social Support, and Environmental Support for Physical Activity in 118 Mexican-origin women. The adjusted regression revealed a significant association between ELP and perceived physical activity self-efficacy (β = 234.2, p = .004), but not with physical activity social support. In South Carolina, CHAMPS leisure-time MVPA (411.4 versus 114.3 minutes, p < .05) was significantly different between women in the high ELP quartile and those in the very low quartile. Among high ELP Mexican-origin women, participants in Texas reported significantly higher MVPA measured by accelerometry (p = .042) than those in South Carolina. Our findings indicate that ELP was associated with physical activity and that contextual factors may also play a role.

Hybrid Elastin-like Polypeptide–Polyethylene Glycol (ELP-PEG) Hydrogels with Improved Transparency and Independent Control of Matrix Mechanics and Cell Ligand Density

PubMed Central

2015-01-01

Hydrogels have been developed as extracellular matrix (ECM) mimics both for therapeutic applications and basic biological studies. In particular, elastin-like polypeptide (ELP) hydrogels, which can be tuned to mimic several biochemical and physical characteristics of native ECM, have been constructed to encapsulate various types of cells to create in vitro mimics of in vivo tissues. However, ELP hydrogels become opaque at body temperature because of ELP’s lower critical solution temperature behavior. This opacity obstructs light-based observation of the morphology and behavior of encapsulated cells. In order to improve the transparency of ELP hydrogels for better imaging, we have designed a hybrid ELP-polyethylene glycol (PEG) hydrogel system that rapidly cross-links with tris(hydroxymethyl) phosphine (THP) in aqueous solution via Mannich-type condensation. As expected, addition of the hydrophilic PEG component significantly improves the light transmittance. Coherent anti-Stokes Raman scattering (CARS) microscopy reveals that the hybrid ELP-PEG hydrogels have smaller hydrophobic ELP aggregates at 37 °C. Importantly, this hydrogel platform enables independent tuning of adhesion ligand density and matrix stiffness, which is desirable for studies of cell–matrix interactions. Human fibroblasts encapsulated in these hydrogels show high viability (>98%) after 7 days of culture. High-resolution confocal microscopy of encapsulated fibroblasts reveals that the cells adopt a more spread morphology in response to higher RGD ligand concentrations and softer gel mechanics. PMID:25111283

ALK1 expression in oral lichen planus: a possible relation to microvessel density.

PubMed

Hazzaa, Hala H A; El-Wakeel, Naglaa M; Attia, Enas A S; Abo Hager, Eman A

2016-05-01

To assess the expression of activin receptor-like kinase 1 (ALK1) and investigate its possible relationship with microvessel density (MVD) in different forms of oral lichen planus (OLP) compared to controls' biopsies. Biopsies from 20 reticular/papular OLP (R/PLP), 20 atrophic/erosive OLP (A/ELP) patients, and 20 healthy subjects were immunohistochemically analyzed and statistically compared and correlated for ALK1 expression and MVD as assessed by CD34 expression. All OLP specimens revealed the presence of positive cytoplasmic CD34 immunostaining in endothelial cells, with statistically high significant MVD in each of R/PLP (Median; M = 4.40) and A/ELP (M = 7.69) compared to controls (M = 1.16) (P < 0.001). Statistically significant MVD was found in A/ELP compared to R/PLP (P < 0.001). All control specimens revealed negative ALK1 immunostaining of the few inflammatory cells found, while 85% of A/ELP cases and 70% of R/PLP cases showed positively immunostained sections for ALK-1, with statistically significant higher ALK1 expression In A/ELP (M = 1.95) compared to R/PLP (M = 0.86) (P = 0.005). No significant correlation between CD34 and ALK1 was detected in R/PLP (r = 0.081), while a barely moderate positive correlation was found in A/ELP (r = 0.396). ALK1 expression and MVD are increased in OLP, particularly in A/ELP type. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Elastin-like Protein-Hyaluronic acid (ELP-HA) Hydrogels with Decoupled Mechanical and Biochemical cues for Cartilage Regeneration

PubMed Central

Zhu, Danqing; Wang, Huiyuan; Trinh, Pavin; Heilshorn, Sarah C.; Yang, Fan

2018-01-01

Hyaluronic acid (HA) is a major component of cartilage extracellular matrix and is an attractive material for use as 3D injectable matrices for cartilage regeneration. While previous studies have shown the promise of HA-based hydrogels to support cell-based cartilage formation, varying HA concentration generally led to simultaneous changes in both biochemical cues and stiffness. How cells respond to the change of biochemical content of HA remains largely unknown. Here we report an adaptable elastin-like protein-hyaluronic acid (ELP-HA) hydrogel platform using dynamic covalent chemistry, which allows varyiation of HA concentration without affecting matrix stiffness. ELP-HA hydrogels were created through dynamic hydrazone bonds via the reaction between hydrazine-modified ELP (ELP-HYD) and aldehyde-modified HA (HA-ALD). By tuning the stoichiometric ratio of aldehyde groups to hydrazine groups while maintaining ELP-HYD concentration constant, hydrogels with variable HA concentration (1.5%, 3%, or 5%) (w/v) were fabricated with comparable stiffness. To evaluate the effects of HA concentration on cell-based cartilage regeneration, chondrocytes were encapsulated within ELP-HA hydrogels with varying HA concentration. Increasing HA concentration led to a dose-dependent increase in cartilage-marker gene expression and enhanced sGAG deposition while minimizing undesirable fibrocartilage phenotype. The use of adaptable protein hydrogels formed via dynamic covalent chemistry may be broadly applicable as 3D scaffolds with decoupled niche properties to guide other desirable cell fates and tissue repair. PMID:28268018

Elastin-like protein-hyaluronic acid (ELP-HA) hydrogels with decoupled mechanical and biochemical cues for cartilage regeneration.

PubMed

Zhu, Danqing; Wang, Huiyuan; Trinh, Pavin; Heilshorn, Sarah C; Yang, Fan

2017-05-01

Hyaluronic acid (HA) is a major component of cartilage extracellular matrix and is an attractive material for use as 3D injectable matrices for cartilage regeneration. While previous studies have shown the promise of HA-based hydrogels to support cell-based cartilage formation, varying HA concentration generally led to simultaneous changes in both biochemical cues and stiffness. How cells respond to the change of biochemical content of HA remains largely unknown. Here we report an adaptable elastin-like protein-hyaluronic acid (ELP-HA) hydrogel platform using dynamic covalent chemistry, which allows variation of HA concentration without affecting matrix stiffness. ELP-HA hydrogels were created through dynamic hydrazone bonds via the reaction between hydrazine-modified ELP (ELP-HYD) and aldehyde-modified HA (HA-ALD). By tuning the stoichiometric ratio of aldehyde groups to hydrazine groups while maintaining ELP-HYD concentration constant, hydrogels with variable HA concentration (1.5%, 3%, or 5%) (w/v) were fabricated with comparable stiffness. To evaluate the effects of HA concentration on cell-based cartilage regeneration, chondrocytes were encapsulated within ELP-HA hydrogels with varying HA concentration. Increasing HA concentration led to a dose-dependent increase in cartilage-marker gene expression and enhanced sGAG deposition while minimizing undesirable fibrocartilage phenotype. The use of adaptable protein hydrogels formed via dynamic covalent chemistry may be broadly applicable as 3D scaffolds with decoupled niche properties to guide other desirable cell fates and tissue repair. Copyright © 2017 Elsevier Ltd. All rights reserved.

Engineered Protein Coatings to Improve the Osseointegration of Dental and Orthopaedic Implants

PubMed Central

Raphel, Jordan; Karlsson, Johan; Galli, Silvia; Wennerberg, Ann; Lindsay, Christopher; Haugh, Matthew; Pajarinen, Jukka; Goodman, Stuart B.; Jimbo, Ryo; Andersson, Martin; Heilshorn, Sarah C.

2016-01-01

Here we present the design of an engineered, elastin-like protein (ELP) that is chemically modified to enable stable coatings on the surfaces of titanium-based dental and orthopaedic implants by novel photocrosslinking and solution processing steps. The ELP includes an extended RGD sequence to confer bio-signaling and an elastin-like sequence for mechanical stability. ELP thin films were fabricated on cp-Ti and Ti6Al4V surfaces using scalable spin and dip coating processes with photoactive covalent crosslinking through a carbene insertion mechanism. The coatings withstood procedures mimicking dental screw and hip replacement stem implantations, a key metric for clinical translation. They promoted rapid adhesion of MG63 osteoblast-like cells, with over 80% adhesion after 24 hours, compared to 38% adhesion on uncoated Ti6Al4V. MG63 cells produced significantly more mineralization on ELP coatings compared to uncoated Ti6Al4V. Human bone marrow mesenchymal stem cells (hMSCs) had an earlier increase in alkaline phosphatase activity, indicating more rapid osteogenic differentiation and mineral deposition on adhesive ELP coatings. Rat tibia and femur in vivo studies demonstrated that cell-adhesive ELP-coated implants increased bone-implant contact area and interfacial strength after one week. These results suggest that ELP coatings withstand surgical implantation and promote rapid osseointegration, enabling earlier implant loading and potentially preventing micromotion that leads to aseptic loosening and premature implant failure. PMID:26790146

Tuning Thermoresponsive Properties of Cationic Elastin-like Polypeptides by Varying Counterions and Side-Chains.

PubMed

Petitdemange, Rosine; Garanger, Elisabeth; Bataille, Laure; Bathany, Katell; Garbay, Bertrand; Deming, Timothy J; Lecommandoux, Sébastien

2017-05-17

We report the synthesis of methionine-containing recombinant elastin-like polypeptides (ELPs) of different lengths that contain periodically spaced methionine residues. These ELPs were chemoselectively alkylated at all methionine residues to give polycationic derivatives. Some of these samples were found to possess solubility transitions in water, where the temperature of these transitions varied with ELP concentration, nature of the methionine alkylating group, and nature of the sulfonium counterions. These studies show that introduction and controlled spacing of methionine sulfonium residues into ELPs can be used as a means both to tune their solubility transition temperatures in water using a variety of different parameters and to introduce new side-chain functionality.

Elementary Teachers' Perceptions of Elementary Principals' Effectiveness

ERIC Educational Resources Information Center

Fridenvalds, Kriss R.

2012-01-01

This dissertation examined the beliefs of elementary teachers to determine if their perceptions of effective principal leadership align to transformational leadership theory vis-a-vis the Educational Leadership Policy Standards (ELPS). A phenomenological, single-case study approach was utilized by means of a mixed-methodological, Web-based survey,…

Biosynthesis of Thermoresponsive Magnetic Nanoparticles by Magnetosome Display System.

PubMed

Yoshino, Tomoko; Shimada, Takumi; Ito, Yasuhito; Honda, Toru; Maeda, Yoshiaki; Matsunaga, Tadashi; Tanaka, Tsuyoshi

2018-05-16

Thermoresponsive magnetic nanoparticles (MNPs) were synthesized using a magnetosome display system. An elastin-like polypeptide decamer of VPGVG (ELP 10 ), which is hydrophobic above the transition temperature ( T t ) and can form an insoluble aggregation, was immobilized on biogenic MNPs in the magnetotactic bacterium, Magnetospirillum magneticum AMB-1. It was suggested that hydrophobicity of the MNP surface increased at 60 °C compared with 20 °C by the immobilization of ELP 10 . Size distribution analysis indicated that the immobilization of ELP 10 onto MNPs induced the increased hydrophobicity with increasing temperatures up to 60 °C, promoting aggregation of the particles by hydrophobic and magnetic interactions. These results suggest that the acceleration of magnetic collection at 60 °C was caused by particle aggregation promoted by hydrophobic interaction between ELP-MNPs. Furthermore, the immobilization of ELP on MNPs gave a quick magnetic collection at 60 °C by external magnetic field. The thermoresponsive properties will further expand the utility of biotechnological applications of biogenic MNPs.

Energy-landscape paving for prediction of face-centered-cubic hydrophobic-hydrophilic lattice model proteins

NASA Astrophysics Data System (ADS)

Liu, Jingfa; Song, Beibei; Liu, Zhaoxia; Huang, Weibo; Sun, Yuanyuan; Liu, Wenjie

2013-11-01

Protein structure prediction (PSP) is a classical NP-hard problem in computational biology. The energy-landscape paving (ELP) method is a class of heuristic global optimization algorithm, and has been successfully applied to solving many optimization problems with complex energy landscapes in the continuous space. By putting forward a new update mechanism of the histogram function in ELP and incorporating the generation of initial conformation based on the greedy strategy and the neighborhood search strategy based on pull moves into ELP, an improved energy-landscape paving (ELP+) method is put forward. Twelve general benchmark instances are first tested on both two-dimensional and three-dimensional (3D) face-centered-cubic (fcc) hydrophobic-hydrophilic (HP) lattice models. The lowest energies by ELP+ are as good as or better than those of other methods in the literature for all instances. Then, five sets of larger-scale instances, denoted by S, R, F90, F180, and CASP target instances on the 3D FCC HP lattice model are tested. The proposed algorithm finds lower energies than those by the five other methods in literature. Not unexpectedly, this is particularly pronounced for the longer sequences considered. Computational results show that ELP+ is an effective method for PSP on the fcc HP lattice model.

Triple selectin knockout (ELP-/-) mice fail to develop OVA-induced acute asthma phenotype

PubMed Central

2011-01-01

Objective The recruitment of leukocytes from circulation to sites of inflammation requires several families of adhesion molecules among which are selectins expressed on a variety of cells. In addition, they have also been shown to play key roles in the activation of cells in inflammation. Methods To explore the collective role of E-, L-, and P- selectins in OVA-induced Th2 mediated response in acute asthma pathophysiology, ELP-/- mice were used and compared with age-matched wildtype (WT). Results Asthma phenotype was assessed by measuring pulmonary function, inflammation and OVA-specific serum IgE, which were completely abrogated in ELP-/- mice. Adoptive transfer of sensitized L selectin+CD4+ T cells into naïve ELP-/- mice which post-OVA challenge, developed asthma, suggesting that L-selectin may be critically involved in the onset of Th2 response in asthma. Tissue resident ELP-deficient cells were otherwise functionally competent as proved by normal proliferative response. Conclusions: Comparative studies between ELP-/- and WT mice uncovered functional roles of these three integrins in inflammatory response in allergic asthma. All three selectins seem to impede inflammatory migration while only L-selectin also possibly regulates activation of specific T cell subsets in lung and airways. PMID:21835035

Stepwise Mechanism of Temperature-Dependent Coacervation of the Elastin-like Peptide Analogue Dimer, (C(WPGVG)3)2.

PubMed

Tatsubo, Daiki; Suyama, Keitaro; Miyazaki, Masaya; Maeda, Iori; Nose, Takeru

2018-03-13

Elastin-like peptides (ELPs) are distinct, repetitive, hydrophobic sequences, such as (VPGVG) n , that exhibit coacervation, the property of reversible, temperature-dependent self-association and dissociation. ELPs can be found in elastin and have been developed as new scaffold biomaterials. However, the detailed relationship between their amino acid sequences and coacervation properties remains obscure because of the structural flexibility of ELPs. In this study, we synthesized a novel, dimeric ELP analogue (H-C(WPGVG) 3 -NH 2 ) 2 , henceforth abbreviated (CW3)2, and analyzed its self-assembly properties and structural factors as indicators of coacervation. Turbidity measurements showed that (CW3)2 demonstrated coacervation at a concentration much lower than that of its monomeric form and another ELP. In addition, the coacervate held water-soluble dye molecules. Thus, potent and distinct coacervation was obtained with a remarkably short sequence of (CW3)2. Furthermore, fluorescence microscopy, dynamic light scattering, and optical microscopy revealed that the coacervation of (CW3)2 was a stepwise process. The structural factors of (CW3)2 were analyzed by molecular dynamics simulations and circular dichroism spectroscopy. These measurements indicated that helical structures primarily consisting of proline and glycine became more disordered at high temperatures with concurrent, significant exposure of their hydrophobic surfaces. This extreme change in the hydrophobic surface contributes to the potent coacervation observed for (CW3)2. These results provide important insights into more efficient applications of ELPs and their analogues, as well as the coacervation mechanisms of ELP and elastin.

Elastin-like polypeptide matrices for enhancing adeno-associated virus-mediated gene delivery to human neural stem cells.

PubMed

Kim, J-S; Chu, H S; Park, K I; Won, J-I; Jang, J-H

2012-03-01

The successful development of efficient and safe gene delivery vectors continues to be a major obstacle to gene delivery in stem cells. In this study, we have developed an elastin-like polypeptide (ELP)-mediated adeno-associated virus (AAV) delivery system for transducing fibroblasts and human neural stem cells (hNSCs). AAVs have significant promise as therapeutic vectors because of their safety and potential for use in gene targeting in stem cell research. ELP has been recently employed as a biologically inspired 'smart' biomaterial that exhibits an inverse temperature phase transition, thereby demonstrating promise as a novel drug carrier. The ELP that was investigated in this study was composed of a repetitive penta-peptide with [Val-Pro-Gly-Val-Gly]. A novel AAV variant, AAV r3.45, which was previously engineered by directed evolution to enhance transduction in rat NSCs, was nonspecifically immobilized onto ELPs that were adsorbed beforehand on a tissue culture polystyrene surface (TCPS). The presence of different ELP quantities on the TCPS led to variations in surface morphology, roughness and wettability, which were ultimately key factors in the modulation of cellular transduction. Importantly, with substantially reduced viral quantities compared with bolus delivery, ELP-mediated AAV delivery significantly enhanced delivery efficiency in fibroblasts and hNSCs, which have great potential for use in tissue engineering applications and neurodegenerative disorder treatments, respectively. The enhancement of cellular transduction in stem cells, as well as the feasibility of ELPs for utilization in three-dimensional scaffolds, will contribute to the advancement of gene therapy for stem cell research and tissue regenerative medicine.

The Design and Delivery of a Thermally Responsive Peptide to Inhibit S100B Mediated Neurodegeneration

PubMed Central

Hearst, Scoty M; Walker, Leslie R; Shao, Qingmei; Lopez, Mariper; Raucher, Drazen; Vig, Parminder J S

2011-01-01

S100B, a glial secreted protein is believed to play a major role in neurodegeneration in Alzheimer's disease, Down syndrome, traumatic brain injury and spinocerebellar ataxia type 1 (SCA1). SCA1 is a trinucleotide repeat disorder in which the expanded polyglutamine mutation in the protein ataxin-1 primarily targets Purkinje cells (PCs) of the cerebellum. Currently, the exact mechanism of S100B mediated PC damage in SCA1 is not clear. However, here we show that S100B may act via the activation of the RAGE signaling pathway resulting in oxidative stress mediated injury to mutant ataxin-1 expressing neurons. To combat S100B mediated neurodegeneration, we have designed a selective thermally responsive S100B inhibitory peptide, Synb1-ELP-TRTK. Our therapeutic polypeptide was developed using three key elements: (1) the elastin-like polypeptide (ELP), a thermally responsive polypeptide, (2) the TRTK12 peptide, a known S100B inhibitory peptide and (3) a cell penetrating peptide, Synb1, to enhance intracellular delivery. Binding studies revealed that our peptide, Synb1-ELP-TRTK, interacts with its molecular target S100B and maintains a high S100B binding affinity as comparable with the TRTK12 peptide alone. In addition, in vitro studies revealed that Synb1-ELP-TRTK treatment reduces S100B uptake in SHSY5Y cells. Furthermore, the Synb1-ELP-TRTK peptide decreased S100B induced oxidative damage to mutant ataxin-1 expressing neurons. To test the delivery capabilities of ELP based therapeutic peptides to the cerebellum; we treated mice with fluorescently labeled Synb1-ELP and observed that thermal targeting enhanced peptide delivery to the cerebellum. Here, we have laid the framework for thermal based therapeutic targeting to regions of the brain, particularly the cerebellum. Overall, our data suggests that thermal targeting of ELP based therapeutic peptides to the cerebellum is a novel treatment strategy for cerebellar neurodegenerative disorders. PMID:21958864

Thermally targeted delivery of chemotherapeutics and anti-cancer peptides by elastin-like polypeptide.

PubMed

Raucher, Drazen; Massodi, Iqbal; Bidwell, Gene L

2008-03-01

Current chemotherapy treatment of solid tumors is limited due to a lack of specific delivery of the drugs to the tumor, leading to systemic toxicity. Therefore, it is necessary to develop targeted cancer therapies and tumor-targeted drug carriers. The authors review the development of elastin-like polypeptide (ELP) as a potential carrier for thermally targeted delivery of therapeutics. The authors searched Medline for articles concerning the application of ELP as a drug delivery vector for small molecule drugs and therapeutic peptides. ELP has been demonstrated to be a promising thermally targeted carrier. Further examination of the in vivo biodistribution and efficacy will provide the necessary data to advance ELP technology toward the ultimate goal of human therapeutics.

Immobilization of R-ω-transaminase on MnO2 nanorods for catalyzing the conversion of (R)-1-phenylethylamine.

PubMed

Sun, Jian; Cui, Wen-Hui; Du, Kun; Gao, Qian; Du, Mengmeng; Ji, Peijun; Feng, Wei

2017-03-10

R-ɷ-transaminases transfer an amino group from an amino donor (e.g. (R)-1-phenylethylamine) onto an amino acceptor (e.g. pyruvate), resulting a co-product (e.g. d-alanine). This work intends to immobilize R-ɷ-Transaminase on MnO 2 nanorods to achieve multienzyme catalysis. R-ɷ-Transaminase (RTA) and d-amino acid oxidase (DAAO) have been fused to an elastin-like polypeptide (ELP) separately through genetic engineering of the enzymes. ELP-RTA and ELP-DAAO have been separately immobilized on polydopamine-coated MnO 2 nanorods. When the two immobilized enzymes were used together in one pot, the transformation of (R)-1-phenylethylamine was catalyzed by the immobilized ELP-RTA, and the co-product d-alanine was converted back to pyruvate under the catalysis of the immobilized ELP-DAAO, achieving the recycling of pyruvate in situ. Thus pyruvate was maintained at a low concentration in order to reduce its negative effect. On the other hand, the generated H 2 O 2 of ELP-DAAO was decomposed by the MnO 2 nanorods, and the evolved oxygen oxidized the reduced cofactors of ELP-DAAO. Forming the circles of hydrogen peroxide→oxygen→hydrogen peroxide accelerated the deamination reaction. The highly efficient conversion of the co-product d-alanine back to pyruvate accelerated the forming of the pyruvate→d-alanine→pyruvate cycle between the two immobilized enzymes. The coordination of the pyruvate→d-alanine→pyruvate and hydrogen peroxide→oxygen→hydrogen peroxide cycles accelerated the transformation of (R)-1-phenylethylamine. As a result, As a result, the immobilized enzymes achieved a conversion of 98±1.8% in comparison to 69.6±1.2% by free enzymes. Copyright © 2017 Elsevier B.V. All rights reserved.

78 FR 55068 - Request for Information To Inform the Title III Evaluation and Research Studies Agenda

Federal Register 2010, 2011, 2012, 2013, 2014

2013-09-09

... DEPARTMENT OF EDUCATION [Docket ID ED-2013-OELA-0117] Request for Information To Inform the Title III Evaluation and Research Studies Agenda AGENCY: Office of English Language Acquisition, Language..., Title III of the ESEA requires States to develop English language proficiency (ELP) standards that are...

Effects of cell penetrating Notch inhibitory peptide conjugated to elastin-like polypeptide on glioblastoma cells.

PubMed

Opačak-Bernardi, Teuta; Ryu, Jung Su; Raucher, Drazen

2017-07-01

Notch pathway was found to be activated in most glioblastomas (GBMs), underlining the importance of Notch in formation and recurrence of GBM. In this study, a Notch inhibitory peptide, dominant negative MAML (dnMAML), was conjugated to elastin-like polypeptide (ELP) for tumor targeted delivery. ELP is a thermally responsive polypeptide that can be actively and passively targeted to the tumor site by localized application of hyperthermia. This complex was further modified with the addition of a cell penetrating peptide, SynB1, for improved cellular uptake and blood-brain barrier penetration. The SynB1-ELP1-dnMAML was examined for its cellular uptake, cytotoxicity, apoptosis, cell cycle inhibition and the inhibition of target genes' expression. SynB1-ELP1-dnMAML inhibited the growth of D54 and U251 cells by inducing apoptosis and cell cycle arrest, especially in the presence of hyperthermia. Hyperthermia increased overall uptake of the polypeptide by the cells and enhanced the resulting pharmacological effects of dnMAML, showing the inhibition of targets of Notch pathway such as Hes-1 and Hey-L. These results confirm that dnMAML is an effective Notch inhibitor and combination with ELP may allow thermal targeting of the SynB1-ELP1-dnMAML complex in cancer cells while avoiding the dangers of systemic Notch inhibition.

The design and delivery of a PKA inhibitory polypeptide to treat SCA1.

PubMed

Hearst, Scoty M; Shao, Qingmei; Lopez, Mariper; Raucher, Drazen; Vig, Parminder J S

2014-10-01

Spinocerebellar ataxia-1 (SCA1) is a neurodegenerative disease that primarily targets Purkinje cells (PCs) of the cerebellum. The exact mechanism of PC degeneration is unknown, however, it is widely believed that mutant ataxin-1 becomes toxic because of the phosphorylation of its serine 776 (S776) residue by cAMP-dependent protein kinase A (PKA). Therefore, to directly modulate mutant ATXN1 S776 phosphorylation and aggregation, we designed a therapeutic polypeptide to inhibit PKA. This polypeptide comprised of a thermally responsive elastin-like peptide (ELP) carrier, which increases peptide half-life, a PKA inhibitory peptide (PKI), and a cell-penetrating peptide (Synb1). We observed that our therapeutic polypeptide, Synb1-ELP-PKI, inhibited PKA activity at concentrations similar to the PKI peptide. Additionally, Synb1-ELP-PKI significantly suppressed mutant ATXN1 S776 phosphorylation and intranuclear inclusion formation in cell culture. Further, Synb1-ELP-PKI treatment improved SCA1 PC morphology in cerebellar slice cultures. Furthermore, the Synb1-ELP peptide carrier crossed the blood-brain barrier and localized to the cerebellum via the i.p. or intranasal route. Here, we show the intranasal delivery of ELP-based peptides to the brain as a novel delivery strategy. We also demonstrate that our therapeutic polypeptide has a great potential to target the neurotoxic S776 phosphorylation pathway in the SCA1 disease. © 2014 International Society for Neurochemistry.

Guanidinium can both Cause and Prevent the Hydrophobic Collapse of Biomacromolecules

PubMed Central

2017-01-01

A combination of Fourier transform infrared and phase transition measurements as well as molecular computer simulations, and thermodynamic modeling were performed to probe the mechanisms by which guanidinium (Gnd+) salts influence the stability of the collapsed versus uncollapsed state of an elastin-like polypeptide (ELP), an uncharged thermoresponsive polymer. We found that the cation’s action was highly dependent upon the counteranion with which it was paired. Specifically, Gnd+ was depleted from the ELP/water interface and was found to stabilize the collapsed state of the macromolecule when paired with well-hydrated anions such as SO42–. Stabilization in this case occurred via an excluded volume (or depletion) effect, whereby SO42– was strongly partitioned away from the ELP/water interface. Intriguingly, at low salt concentrations, Gnd+ was also found to stabilize the collapsed state of the ELP when paired with SCN–, which is a strong binder for the ELP. In this case, the anion and cation were both found to be enriched in the collapsed state of the polymer. The collapsed state was favored because the Gnd+ cross-linked the polymer chains together. Moreover, the anion helped partition Gnd+ to the polymer surface. At higher salt concentrations (>1.5 M), GndSCN switched to stabilizing the uncollapsed state because a sufficient amount of Gnd+ and SCN– partitioned to the polymer surface to prevent cross-linking from occurring. Finally, in a third case, it was found that salts which interacted in an intermediate fashion with the polymer (e.g., GndCl) favored the uncollapsed conformation at all salt concentrations. These results provide a detailed, molecular-level, mechanistic picture of how Gnd+ influences the stability of polypeptides in three distinct physical regimes by varying the anion. It also helps explain the circumstances under which guanidinium salts can act as powerful and versatile protein denaturants. PMID:28054487

Conserved genetic determinant of motor organ identity in Medicago truncatula and related legumes

PubMed Central

Chen, Jianghua; Moreau, Carol; Liu, Yu; Kawaguchi, Masayoshi; Hofer, Julie; Ellis, Noel; Chen, Rujin

2012-01-01

Plants exhibit various kinds of movements that have fascinated scientists and the public for centuries. Physiological studies in plants with the so-called motor organ or pulvinus suggest that cells at opposite sides of the pulvinus mediate leaf or leaflet movements by swelling and shrinking. How motor organ identity is determined is unknown. Using a genetic approach, we isolated a mutant designated elongated petiolule1 (elp1) from Medicago truncatula that fails to fold its leaflets in the dark due to loss of motor organs. Map-based cloning indicated that ELP1 encodes a putative plant-specific LOB domain transcription factor. RNA in situ analysis revealed that ELP1 is expressed in primordial cells that give rise to the motor organ. Ectopic expression of ELP1 resulted in dwarf plants with petioles and rachises reduced in length, and the epidermal cells gained characteristics of motor organ epidermal cells. By identifying ELP1 orthologs from other legume species, namely pea (Pisum sativum) and Lotus japonicus, we show that this motor organ identity is regulated by a conserved molecular mechanism. PMID:22689967

Sustained release of antibiotics from injectable and thermally responsive polypeptide depots.

PubMed

Adams, Samuel B; Shamji, Mohammed F; Nettles, Dana L; Hwang, Priscilla; Setton, Lori A

2009-07-01

Biodegradable polymeric scaffolds are of interest for delivering antibiotics to local sites of infection in orthopaedic applications, such as bone and diarthrodial joints. The objective of this study was to develop a biodegradable scaffold with ease of drug loading in aqueous solution, while providing for drug depot delivery via syringe injection. Elastin-like polypeptides (ELPs) were used for this application, biopolymers of repeating pentapeptide sequences that were thermally triggered to undergo in situ depot formation at body temperature. ELPs were modified to enable loading with the antibiotics, cefazolin, and vancomycin, followed by induction of the phase transition in vitro. Cefazolin and vancomycin concentrations were monitored, as well as bioactivity of the released antibiotics, to test an ability of the ELP depot to provide for prolonged release of bioactive drugs. Further tests of formulation viscosity were conducted to test suitability as an injectable drug carrier. Results demonstrate sustained release of therapeutic concentrations of bioactive antibiotics by the ELP, with first-order time constants for drug release of approximately 25 h for cefazolin and approximately 500 h for vancomycin. These findings illustrate that an injectable, in situ forming ELP depot can provide for sustained release of antibiotics with an effect that varies across antibiotic formulation. ELPs have important advantages for drug delivery, as they are known to be biocompatible, biodegradable, and elicit no known immune response. These benefits suggest distinct advantages over currently used carriers for antibiotic drug delivery in orthopedic applications. (c) 2008 Wiley Periodicals, Inc.

Phosphorylation of Elp1 by Hrr25 Is Required for Elongator-Dependent tRNA Modification in Yeast

PubMed Central

Abdel-Fattah, Wael; Jablonowski, Daniel; Di Santo, Rachael; Thüring, Kathrin L.; Scheidt, Viktor; Hammermeister, Alexander; ten Have, Sara; Helm, Mark; Schaffrath, Raffael; Stark, Michael J. R.

2015-01-01

Elongator is a conserved protein complex comprising six different polypeptides that has been ascribed a wide range of functions, but which is now known to be required for modification of uridine residues in the wobble position of a subset of tRNAs in yeast, plants, worms and mammals. In previous work, we showed that Elongator's largest subunit (Elp1; also known as Iki3) was phosphorylated and implicated the yeast casein kinase I Hrr25 in Elongator function. Here we report identification of nine in vivo phosphorylation sites within Elp1 and show that four of these, clustered close to the Elp1 C-terminus and adjacent to a region that binds tRNA, are important for Elongator's tRNA modification function. Hrr25 protein kinase directly modifies Elp1 on two sites (Ser-1198 and Ser-1202) and through analyzing non-phosphorylatable (alanine) and acidic, phosphomimic substitutions at Ser-1198, Ser-1202 and Ser-1209, we provide evidence that phosphorylation plays a positive role in the tRNA modification function of Elongator and may regulate the interaction of Elongator both with its accessory protein Kti12 and with Hrr25 kinase. PMID:25569479

Elastin-like polypeptides: Therapeutic applications for an emerging class of nanomedicines.

PubMed

Despanie, Jordan; Dhandhukia, Jugal P; Hamm-Alvarez, Sarah F; MacKay, J Andrew

2016-10-28

Elastin-like polypeptides (ELPs) constitute a genetically engineered class of 'protein polymers' derived from human tropoelastin. They exhibit a reversible phase separation whereby samples remain soluble below a transition temperature (T t ) but form amorphous coacervates above T t . Their phase behavior has many possible applications in purification, sensing, activation, and nanoassembly. As humanized polypeptides, they are non-immunogenic, substrates for proteolytic biodegradation, and can be decorated with pharmacologically active peptides, proteins, and small molecules. Recombinant synthesis additionally allows precise control over ELP architecture and molecular weight, resulting in protein polymers with uniform physicochemical properties suited to the design of multifunctional biologics. As such, ELPs have been employed for various uses including as anti-cancer agents, ocular drug delivery vehicles, and protein trafficking modulators. This review aims to offer the reader a catalogue of ELPs, their various applications, and potential for commercialization across a broad spectrum of fields. Copyright © 2015. Published by Elsevier B.V.

A non-chromatographic protein purification strategy using Src 3 homology domains as generalized capture domains.

PubMed

Kim, Heejae; Chen, Wilfred

2016-09-20

Protein purification using inverse phase transition of elastin-like polypeptide (ELP) domains is a useful alternative to chromatography. Genetic fusions of ELP domains to various proteins have the ability to reversibly transition between soluble monomers and micron-sized aggregates and this has been used to selectively purify many ELP fusions. Affinity domains can enhance this technology by using specific protein binding domains to enable ELP mediated affinity capture (EMAC) of proteins of interest (POI) that have been fused to corresponding affinity ligands. In this paper, we highlight the use of Src homology 3 (SH3) domains and corresponding peptide ligands in EMAC that have differential binding affinities towards SH3 for efficient capture and elution of proteins. Furthermore, differences between capture and elution of a monomeric and a multimeric protein were also studied. Copyright © 2016 Elsevier B.V. All rights reserved.

An incidental enterocolic lymphocytic phlebitis pattern is seen commonly in the rectal stump of patients with diversion colitis superimposed on inflammatory bowel disease.

PubMed

Chetty, R; Hafezi, S; Montgomery, E

2009-05-01

Enterocolic lymphocytic phlebitis (ELP) is an uncommon cause of bowel pathology and most frequently results in ischaemia. It is characterised by an artery-sparing, venulocentric lymphoid infiltrate that causes a phlebitis and vascular compromise. Rare cases of ELP have been encountered with lymphocytic colitis in the absence of ischaemic bowel change. The present study examined the occurrence of ELP in the setting of diversion colitis and inflammatory bowel disease, as well as in random colectomy specimens. The study cohort comprised the following: 26 completion proctectomy specimens for ulcerative colitis with superimposed diversion colitis in the rectal stump; 3 colectomy specimens for Crohn disease with diversion colitis; 6 colectomy specimens for adenocarcinoma and/or diverticular disease with diversion colitis; 34 resection specimens with ulcerative colitis only; 19 with Crohn disease only; and 100 random colon resection specimens for adenocarcinoma, adenoma, diverticular disease and ischaemia. ELP was present in 18 of the 26 ulcerative colitis cases with diversion colitis, 3/3 Crohn disease cases with diversion colitis, 1/6 cases of diverticular disease with diversion colitis, 6/34 cases of ulcerative colitis without diversion, 2/19 Crohn disease cases without diversion colitis, and only 1 of 100 colectomy cases without inflammatory bowel disease or diversion colitis. ELP occurs most frequently in cases that have been diverted for inflammatory bowel disease. Fewer cases of ELP were noted in cases of inflammatory bowel disease in the absence of diversion colitis. It is postulated that altered bowel flora and immune dysregulation may be pivotal in the causation of this association.

Purification of Microbially Expressed Recombinant Proteins via a Dual ELP Split Intein System.

PubMed

Shi, Changhua; Han, Tzu-Chiang; Wood, David W

2017-01-01

Fusions of elastin-like peptide (ELP) purification tags and self-cleaving inteins provide a powerful platform for purifying tagless recombinant proteins without the need for conventional packed-bed columns. A drawback to this method has been premature cleaving of the ELP tag during expression, before the purification procedure can take place. Here we demonstrate a split-intein method, where the self-cleaving intein is divided into two inactive segments during expression and purification. Spontaneous assembly of the purified intein segments then restores self-cleaving activity to deliver the tagless target protein.

Generation of polypeptide-templated gold nanoparticles using ionizing radiation.

PubMed

Walker, Candace Rae; Pushpavanam, Karthik; Nair, Divya Geetha; Potta, Thrimoorthy; Sutiyoso, Caesario; Kodibagkar, Vikram D; Sapareto, Stephen; Chang, John; Rege, Kaushal

2013-08-13

Ionizing radiation, including γ rays and X-rays, are high-energy electromagnetic radiation with diverse applications in nuclear energy, astrophysics, and medicine. In this work, we describe the use of ionizing radiation and cysteine-containing elastin-like polypeptides (C(n)ELPs, where n = 2 or 12 cysteines in the polypeptide sequence) for the generation of gold nanoparticles. In the presence of C(n)ELPs, ionizing radiation doses higher than 175 Gy resulted in the formation of maroon-colored gold nanoparticle dispersions, with maximal absorbance at 520 nm, from colorless metal salts. Visible color changes were not observed in any of the control systems, indicating that ionizing radiation, gold salt solution, and C(n)ELPs were all required for nanoparticle formation. The hydrodynamic diameters of nanoparticles, determined using dynamic light scattering, were in the range of 80-150 nm, while TEM imaging indicated the formation of gold cores 10-20 nm in diameter. Interestingly, C2ELPs formed 1-2 nm diameter gold nanoparticles in the absence of radiation. Our results describe a facile method of nanoparticle formation in which nanoparticle size can be tailored based on radiation dose and C(n)ELP type. Further improvements in these polypeptide-based systems can lead to colorimetric detection of ionizing radiation in a variety of applications.

Micellar Self-Assembly of Recombinant Resilin-/Elastin-Like Block Copolypeptides.

PubMed

Weitzhandler, Isaac; Dzuricky, Michael; Hoffmann, Ingo; Garcia Quiroz, Felipe; Gradzielski, Michael; Chilkoti, Ashutosh

2017-08-14

Reported here is the synthesis of perfectly sequence defined, monodisperse diblock copolypeptides of hydrophilic elastin-like and hydrophobic resilin-like polypeptide blocks and characterization of their self-assembly as a function of structural parameters by light scattering, cryo-TEM, and small-angle neutron scattering. A subset of these diblock copolypeptides exhibit lower critical solution temperature and upper critical solution temperature phase behavior and self-assemble into spherical or cylindrical micelles. Their morphologies are dictated by their chain length, degree of hydrophilicity, and hydrophilic weight fraction of the ELP block. We find that (1) independent of the length of the corona-forming ELP block there is a minimum threshold in the length of the RLP block below which self-assembly does not occur, but that once that threshold is crossed, (2) the RLP block length is a unique molecular parameter to independently tune self-assembly and (3) increasing the hydrophobicity of the corona-forming ELP drives a transition from spherical to cylindrical morphology. Unlike the self-assembly of purely ELP-based block copolymers, the self-assembly of RLP-ELPs can be understood by simple principles of polymer physics relating hydrophilic weight fraction and polymer-polymer and polymer-solvent interactions to micellar morphology, which is important as it provides a route for the de novo design of desired nanoscale morphologies from first principles.

Conformational dynamics of minimal elastin-like polypeptides: the role of proline revealed by molecular dynamics and nuclear magnetic resonance.

PubMed

Glaves, Rachel; Baer, Marcel; Schreiner, Eduard; Stoll, Raphael; Marx, Dominik

2008-12-22

Previous molecular dynamics studies of the elastin-like peptide (ELP) GVG(VPGVG) predict that this ELP undergoes a conformational transition from an open to a more compact closed state upon an increase in temperature. These structural changes occurring in this minimal elastin model at the so-called inverse temperature transition (ITT), which takes place when elastin is heated to temperatures of about 20-40 (omicron)C, are investigated further in this work by means of a combined theoretical and experimental approach. To do this, additional extensive classical molecular dynamics (MD) simulations of the capped octapeptide are carried out, analyzed, and compared to data obtained from homonuclear magnetic resonance (NMR) spectroscopy of the same octapeptide. Moreover, in the previous simulations, the proline residue in the ELP is found to act as a hinge, thereby allowing for the large-amplitude opening and closing conformational motion of the ITT. To explore the role of proline in such elastin repeating units, a point mutant (P5I), which replaces the proline residue with an isoleucine residue, is also investigated using the aforementioned theoretical and experimental techniques. The results show that the site-directed mutation completely alters the properties of this ELP, thus confirming the importance of the highly conserved proline residue in the ITT. Furthermore, a correlation between the two different methods employed is seen. Both methods predict the mutant ELP to be present in an unstructured form and the wild type ELP to have a beta-turn-like structure. Finally, the role of the peptidyl cis to trans isomerization of the proline hinge is assessed in detail.

Structural Model for the Flip-Flop Action in Thiamin Pyrophosphate-Dependent Human Pyruvate Dehydrogenase

NASA Technical Reports Server (NTRS)

Ciszak, Ewa; Dominiak, Paulina

2003-01-01

The derivative of vitamin B1 thiamin pyrophosphate (TPP) is a cofactor of enzymes performing catalysis in pathways of energy production, including (i) decarboxylation of alpha-keto acids followed by (ii) transketolation. These enzymes have shown a common mechanism of TPP activation by imposing an active V-conformation of this coenzyme that brings the N4 atom of the aminopyrimidine ring to the distance required for the intramolecular C-H N hydrogen-bonding with the C2- atom of the thiazolium ring. The reactive C2 atom of TPP is the nucleophile that attacks the carbonyl carbon of different substrates used by the TPP-dependent enzymes. The structure of the heterotetrameric human pyruvate dehydrogenase (Elp) recently determined in our laboratory (1) revealed the association pattern of the subunits and the specifics of two chemically equivalent cofactor binding sites. Dynamic nonequivalence of these two cofactor sites directs the flip-flop action of this enzyme, depending upon which two active sites effect each other (2). The crystal structure derived from the holo-form of Elp provided the basis for the model of the flip-flop action of Elp in which different steps of the catalytic reaction are performed in each of the two cofactor sites at any given moment, where these steps are governed by the concerted shuttle-like motion of the subunits. It is further proposed that balancing a hydrogen-bond network and related cofactor geometry determine the continuity of catalytic events.

78 FR 19711 - Center for Devices and Radiological Health: Experiential Learning Program

Federal Register 2010, 2011, 2012, 2013, 2014

2013-04-02

... Program (ELP). The ELP provides a formal training mechanism for regulatory review staff to visit research... medical device establishments, including, research, manufacturing, academia, and health care facilities.... Clinical use of orthopedic bone void Observation of surgical filler devices. procedures (posterolateral...

Hydration and conformational mechanics of single, end-tethered elastin-like polypeptides.

PubMed

Valiaev, Alexei; Lim, Dong Woo; Schmidler, Scott; Clark, Robert L; Chilkoti, Ashutosh; Zauscher, Stefan

2008-08-20

We investigated the effect of temperature, ionic strength, solvent polarity, and type of guest residue on the force-extension behavior of single, end-tethered elastin-like polypeptides (ELPs), using single molecule force spectroscopy (SMFS). ELPs are stimulus-responsive polypeptides that contain repeats of the five amino acids Val-Pro-Gly-Xaa-Gly (VPGXG), where Xaa is a guest residue that can be any amino acid with the exception of proline. We fitted the force-extension data with a freely jointed chain (FJC) model which allowed us to resolve small differences in the effective Kuhn segment length distributions that largely arise from differences in the hydrophobic hydration behavior of ELP. Our results agree qualitatively with predictions from recent molecular dynamics simulations and demonstrate that hydrophobic hydration modulates the molecular elasticity for ELPs. Furthermore, our results show that SMFS, when combined with our approach for data analysis, can be used to study the subtleties of polypeptide-water interactions and thus provides a basis for the study of hydrophobic hydration in intrinsically unstructured biomacromolecules.

Molecular Mobility in Hyperbranched Polymers and Their Interaction with an Epoxy Matrix

PubMed Central

Román, Frida; Colomer, Pere; Calventus, Yolanda; Hutchinson, John M.

2016-01-01

The molecular mobility related to the glass transition and secondary relaxations in a hyperbranched polyethyleneimine, HBPEI, and its relaxation behaviour when incorporated into an epoxy resin matrix are investigated by dielectric relaxation spectroscopy (DRS) and dynamic mechanical analysis (DMA). Three systems are analysed: HBPEI, epoxy and an epoxy/HBPEI mixture, denoted ELP. The DRS behaviour is monitored in the ELP system in three stages: prior to curing, during curing, and in the fully cured system. In the stage prior to curing, DRS measurements show three dipolar relaxations: γ, β and α, for all systems (HBPEI, epoxy and ELP). The α-relaxation for the ELP system deviates significantly from that for HBPEI, but superposes on that for the epoxy resin. The fully cured thermoset displays both β- and α-relaxations. In DMA measurements, both α- and β-relaxations are observed in all systems and in both the uncured and fully cured systems, similar to the behaviour identified by DRS. PMID:28773319

The Saccharomyces cerevisiae anaphase-promoting complex interacts with multiple histone-modifying enzymes to regulate cell cycle progression.

PubMed

Turner, Emma L; Malo, Mackenzie E; Pisclevich, Marnie G; Dash, Megan D; Davies, Gerald F; Arnason, Terra G; Harkness, Troy A A

2010-10-01

The anaphase-promoting complex (APC), a large evolutionarily conserved ubiquitin ligase complex, regulates cell cycle progression through mitosis and G(1). Here, we present data suggesting that APC-dependent cell cycle progression relies on a specific set of posttranslational histone-modifying enzymes. Multiple APC subunit mutants were impaired in total and modified histone H3 protein content. Acetylated H3K56 (H3K56(Ac)) levels were as reduced as those of total H3, indicating that loading histones with H3K56(Ac) is unaffected in APC mutants. However, under restrictive conditions, H3K9(Ac) and dimethylated H3K79 (H3K79(me2)) levels were more greatly reduced than those of total H3. In a screen for histone acetyltransferase (HAT) and histone deacetylase (HDAC) mutants that genetically interact with the apc5(CA) (chromatin assembly) mutant, we found that deletion of GCN5 or ELP3 severely hampered apc5(CA) temperature-sensitive (ts) growth. Further analyses showed that (i) the elp3Δ gcn5Δ double mutant ts defect was epistatic to that observed in apc5(CA) cells; (ii) gcn5Δ and elp3Δ mutants accumulate in mitosis; and (iii) turnover of the APC substrate Clb2 is not impaired in elp3Δ gcn5Δ cells. Increased expression of ELP3 and GCN5, as well as genes encoding the HAT Rtt109 and the chromatin assembly factors Msi1 and Asf1, suppressed apc5(CA) defects, while increased APC5 expression partially suppressed elp3Δ gcn5Δ growth defects. Finally, we demonstrate that Gcn5 is unstable during G(1) and following G(1) arrest and is stabilized in APC mutants. We present our working model in which Elp3/Gcn5 and the APC work together to facilitate passage through mitosis and G(1). To progress into S, we propose that at least Gcn5 must then be targeted for degradation in an APC-dependent fashion.

English Language Proficiency and Employability Framework for Australian Higher Education Institutions

ERIC Educational Resources Information Center

Arkoudis, Sophie; Baik, Chi; Bexley, Emmaline; Doughney, Lachlan

2014-01-01

This report presents the "English Language Proficiency" (ELP) and Employability Framework", which has been designed to inform and support higher education institutions' (HEIs) policies and practices on ELP and graduate employability. The "Framework" was developed through a review of the national and international…

Perspectives from the European Language Portfolio: Learner Autonomy and Self-Assessment

ERIC Educational Resources Information Center

Kuhn, Barbel, Ed.; Cavana, Maria Luisa Perez, Ed.

2012-01-01

Using constructivist principles and autonomous learning techniques the ELP has pioneered innovative and cutting edge approaches to learning languages that can be applied to learning across the spectrum. Although articles on the success of the ELP project have appeared in some academic journals, "Perspectives from the European Language…

Report of the Committee on the Extended University.

ERIC Educational Resources Information Center

Ohio Univ., Athens.

To offer extended learning opportunities to persons without access to traditional residential education is the purpose of the proposed extended university of Ohio University. This new academic unit would be responsible for the development and administration of the Extended Learning Program (ELP). The ELP admits students who have successfully…

Using the ELP as a Basis for Self- and Peer Assessment When Selecting "Best" Work in Modern-Language Degree Programmes

ERIC Educational Resources Information Center

Dalziel, Fiona; Davies, Gillian; Han, Amy

2016-01-01

The European Language Portfolio (ELP) was designed as a tool that "supports reflective learning and fosters the development of learner autonomy" (Little 2009, "The European Language Portfolio: Where pedagogy and assessment meet". Strasbourg: Council of Europe.…

Enterocolic lymphocytic phlebitis: an unusual cause of abdominal complaints.

PubMed

Gałązka, Krystyna; Tokarek, Tomasz; Gach, Tomasz; Szpor, Joanna

2012-03-01

Enterocolic lymphocytic phlebitis (ELP) is a rare disease of unknown etiology involving most often the intramural and mesenteric small and medium-sized veins of the gastrointestinal tract. The diagnosis of the disorder is based on the histopathological examination of a surgical specimen as endoscopically obtained diagnostic material is usually too superficial. Clinical manifestation of ELP most frequently is characterized by acute symptoms, such as acute abdomen, signs suggesting acute appendicitis, gastrointestinal hemorrhage, sometimes it manifests as chronic gastrointestinal complaints. We report, to our knowledge for the first time in Poland, a case of ELP with clinical symptoms pointing to acute appendicitis, on laparoscopy manifesting as a tumorous mass in the colonic wall with an unchanged appendix.

The Effects of English Vocabulary Mastery on Geometry Achievement

ERIC Educational Resources Information Center

Rockhold, Jonas

2013-01-01

This study examines socioeconomic status (SES), English language proficiency (ELP), vocabulary proficiency (VP), and math proficiency (MP) of students to determine if SES, as determined by the Free and Reduced Lunch Program (FRL), as well as English language proficiency (ELP) and vocabulary proficiency (VP), as measured by CELLA scale scores and…

Development of Intra-Articular Drug Delivery to Alter Progression of Arthritis Following Joint Injury

DTIC Science & Technology

2013-04-01

sTNFRII in combination. The treatments were encapsulated in elastin -like polypeptide (ELP) drug depots that have been associated with a 25-fold...or both IL-1Ra and sTNFRII. The drugs were encapsulated in elastin -like polypeptide (ELP) drug depots that slowly disaggregate for prolonged IA

Practice versus Politics in Danish Day-Care Centres: How to Bridge the Gap in Early Learning?

ERIC Educational Resources Information Center

Clasen, Line Engel; Jensen de López, Kristine

2016-01-01

It is essential that early educators in day-care services possess adequate pedagogical tools for supporting children's communicative development. Early literacy programmes (ELPs) are potential tools. However, studies investigating the effects of ELPs seldom address implementation processes or the programme users' perspectives. This study sheds…

Molecular and Functional Characterization of Lymphoid Progenitor Subsets Reveals a Bipartite Architecture of Human Lymphopoiesis.

PubMed

Alhaj Hussen, Kutaiba; Vu Manh, Thien-Phong; Guimiot, Fabien; Nelson, Elisabeth; Chabaane, Emna; Delord, Marc; Barbier, Maxime; Berthault, Claire; Dulphy, Nicolas; Alberdi, Antonio José; Burlen-Defranoux, Odile; Socié, Gerard; Bories, Jean Christophe; Larghero, Jerôme; Vanneaux, Valérie; Verhoeyen, Els; Wirth, Thierry; Dalod, Marc; Gluckman, Jean Claude; Cumano, Ana; Canque, Bruno

2017-10-17

The classical model of hematopoiesis established in the mouse postulates that lymphoid cells originate from a founder population of common lymphoid progenitors. Here, using a modeling approach in humanized mice, we showed that human lymphoid development stemmed from distinct populations of CD127 - and CD127 + early lymphoid progenitors (ELPs). Combining molecular analyses with in vitro and in vivo functional assays, we demonstrated that CD127 - and CD127 + ELPs emerged independently from lympho-mono-dendritic progenitors, responded differently to Notch1 signals, underwent divergent modes of lineage restriction, and displayed both common and specific differentiation potentials. Whereas CD127 - ELPs comprised precursors of T cells, marginal zone B cells, and natural killer (NK) and innate lymphoid cells (ILCs), CD127 + ELPs supported production of all NK cell, ILC, and B cell populations but lacked T potential. On the basis of these results, we propose a "two-family" model of human lymphoid development that differs from the prevailing model of hematopoiesis. Copyright © 2017 Elsevier Inc. All rights reserved.

Elongator subunit 3 positively regulates plant immunity through its histone acetyltransferase and radical S-adenosylmethionine domains

PubMed Central

2013-01-01

Background Pathogen infection triggers a large-scale transcriptional reprogramming in plants, and the speed of this reprogramming affects the outcome of the infection. Our understanding of this process has significantly benefited from mutants that display either delayed or accelerated defense gene induction. In our previous work we demonstrated that the Arabidopsis Elongator complex subunit 2 (AtELP2) plays an important role in both basal immunity and effector-triggered immunity (ETI), and more recently showed that AtELP2 is involved in dynamic changes in histone acetylation and DNA methylation at several defense genes. However, the function of other Elongator subunits in plant immunity has not been characterized. Results In the same genetic screen used to identify Atelp2, we found another Elongator mutant, Atelp3-10, which mimics Atelp2 in that it exhibits a delay in defense gene induction following salicylic acid treatment or pathogen infection. Similarly to AtELP2, AtELP3 is required for basal immunity and ETI, but not for systemic acquired resistance (SAR). Furthermore, we demonstrate that both the histone acetyltransferase and radical S-adenosylmethionine domains of AtELP3 are essential for its function in plant immunity. Conclusion Our results indicate that the entire Elongator complex is involved in basal immunity and ETI, but not in SAR, and support that Elongator may play a role in facilitating the transcriptional induction of defense genes through alterations to their chromatin. PMID:23856002

Elongator subunit 3 positively regulates plant immunity through its histone acetyltransferase and radical S-adenosylmethionine domains.

PubMed

Defraia, Christopher T; Wang, Yongsheng; Yao, Jiqiang; Mou, Zhonglin

2013-07-16

Pathogen infection triggers a large-scale transcriptional reprogramming in plants, and the speed of this reprogramming affects the outcome of the infection. Our understanding of this process has significantly benefited from mutants that display either delayed or accelerated defense gene induction. In our previous work we demonstrated that the Arabidopsis Elongator complex subunit 2 (AtELP2) plays an important role in both basal immunity and effector-triggered immunity (ETI), and more recently showed that AtELP2 is involved in dynamic changes in histone acetylation and DNA methylation at several defense genes. However, the function of other Elongator subunits in plant immunity has not been characterized. In the same genetic screen used to identify Atelp2, we found another Elongator mutant, Atelp3-10, which mimics Atelp2 in that it exhibits a delay in defense gene induction following salicylic acid treatment or pathogen infection. Similarly to AtELP2, AtELP3 is required for basal immunity and ETI, but not for systemic acquired resistance (SAR). Furthermore, we demonstrate that both the histone acetyltransferase and radical S-adenosylmethionine domains of AtELP3 are essential for its function in plant immunity. Our results indicate that the entire Elongator complex is involved in basal immunity and ETI, but not in SAR, and support that Elongator may play a role in facilitating the transcriptional induction of defense genes through alterations to their chromatin.

Soluble expression and purification of the recombinant bioactive peptide precursor BPP-1 in Escherichia coli using a cELP-SUMO dual fusion system.

PubMed

Rao, Shengqi; Zang, Xiangyu; Yang, Zhenquan; Gao, Lu; Yin, Yongqi; Fang, Weiming

2016-02-01

A bioactive peptide precursor (BPP-1, 14.3 kDa/115AA), a newly designed polypeptide that may exert a potential antihypertensive effect in vivo, is composed of many different ACE inhibitory peptides and antioxidant peptides tandemly linked according to the restriction sites of gastrointestinal proteases. In this report, we present a novel method to obtain soluble BPP-1 in Escherichia coli using cationic elastin-like polypeptide and SUMO (cELP-SUMO) tags. The cELP-SUMO-tagged fusion protein was expressed in soluble form at 20 °C for 20 h. After purification based on the inverse transition cycling (ITC) method, the purified cELP-SUMO-CFPP fusion protein was subsequently cleaved by a SUMO protease to release the mature BPP-1. After a subsequent simple salt precipitation process, approximately 167.2 mg of recombinant BPP-1 was obtained from 1 l of bacterial culture with at least 92% purity. The molecular mass (Mr) of the recombinant BPP-1 was confirmed by MALDI-TOF MS to equal 14,347. The purified BPP-1 was subjected to simulated gastrointestinal digestion, and the resulting hydrolysates exhibited notable ACE inhibitory and antioxidant activities in vitro. This report provides the first description of the soluble production of a bioactive peptide multimer with potential ACE inhibitory and antioxidant activities in E. coli using a cELP-SUMO tag. Copyright © 2015 Elsevier Inc. All rights reserved.

Endotoxin Exposure during Sensitization to Blomia tropicalis Allergens Shifts TH2 Immunity Towards a TH17-Mediated Airway Neutrophilic Inflammation: Role of TLR4 and TLR2

PubMed Central

Barboza, Renato; Câmara, Niels Olsen Saraiva; Gomes, Eliane; Sá-Nunes, Anderson; Florsheim, Esther; Mirotti, Luciana; Labrada, Alexis; Alcântara-Neves, Neuza Maria; Russo, Momtchilo

2013-01-01

Experimental evidence and epidemiological studies indicate that exposure to endotoxin lipopolysaccharide (eLPS) or other TLR agonists prevent asthma. We have previously shown in the OVA-model of asthma that eLPS administration during alum-based allergen sensitization blocked the development of lung TH2 immune responses via MyD88 pathway and IL-12/IFN-γ axis. In the present work we determined the effect of eLPS exposure during sensitization to a natural airborne allergen extract derived from the house dust mite Blomia tropicalis (Bt). Mice were subcutaneously sensitized with Bt allergens co-adsorbed onto alum with or without eLPS and challenged twice intranasally with Bt. Cellular and molecular parameters of allergic lung inflammation were evaluated 24 h after the last Bt challenge. Exposure to eLPS but not to ultrapure LPS (upLPS) preparation during sensitization to Bt allergens decreased the influx of eosinophils and increased the influx of neutrophils to the airways. Inhibition of airway eosinophilia was not observed in IFN-γdeficient mice while airway neutrophilia was not observed in IL-17RA-deficient mice as well in mice lacking MyD88, CD14, TLR4 and, surprisingly, TLR2 molecules. Notably, exposure to a synthetic TLR2 agonist (PamCSK4) also induced airway neutrophilia that was dependent on TLR2 and TLR4 molecules. In the OVA model, exposure to eLPS or PamCSK4 suppressed OVA-induced airway inflammation. Our results suggest that B. tropicalis allergens engage TLR4 that potentiates TLR2 signaling. This dual TLR activation during sensitization results in airway neutrophilic inflammation associated with increased frequency of lung TH17 cells. Our work highlight the complex interplay between bacterial products, house dust mite allergens and TLR signaling in the induction of different phenotypes of airway inflammation. PMID:23805294

Predicting the Fluid-Phase Behavior of Aqueous Solutions of ELP (VPGVG) Sequences Using SAFT-VR.

PubMed

Zhao, Binwu; Lindeboom, Tom; Benner, Steven; Jackson, George; Galindo, Amparo; Hall, Carol K

2017-10-24

The statistical associating fluid theory for potentials of variable range (SAFT-VR) is used to predict the fluid phase behavior of elastin-like polypeptide (ELP) sequences in aqueous solution with special focus on the loci of lower critical solution temperatures (LCSTs). A SAFT-VR model for these solutions is developed following a coarse-graining approach combining information from atomistic simulations and from previous SAFT models for previously reported relevant systems. Constant-pressure temperature-composition phase diagrams are determined for solutions of (VPGVG) n sequences + water with n = 1 to 300. The SAFT-VR equation of state lends itself to the straightforward calculation of phase boundaries so that complete fluid-phase equilibria can be calculated efficiently. A broad range of thermodynamic conditions of temperature and pressure are considered, and regions of vapor-liquid and liquid-liquid coexistence, including LCSTs, are found. The calculated phase boundaries at low concentrations match those measured experimentally. The temperature-composition phase diagrams of the aqueous ELP solutions at low pressure (0.1 MPa) are similar to those of types V and VI phase behavior in the classification of Scott and van Konynenburg. An analysis of the high-pressure phase behavior confirms, however, that a closed-loop liquid-liquid immiscibility region, separate from the gas-liquid envelope, is present for aqueous solutions of (VPGVG) 30 ; such a phase diagram is typical of type VI phase behavior. ELPs with shorter lengths exhibit both liquid-liquid and gas-liquid regions, both of which become less extensive as the chain length of the ELP is decreased. The strength of the hydrogen-bonding interaction is also found to affect the phase diagram of the (VPGVG) 30 system in that the liquid-liquid and gas-liquid regions expand as the hydrogen-bonding strength is decreased and shrink as it is increased. The LCSTs of the mixtures are seen to decrease as the ELP chain length is increased.

Conceptualizing Accessibility for English Language Proficiency Assessments. Research Report. ETS RR-16-07

ERIC Educational Resources Information Center

Guzman-Orth, Danielle; Laitusis, Cara; Thurlow, Martha; Christensen, Laurene

2016-01-01

This paper is the second in a series from Educational Testing Service (ETS) that conceptualizes next-generation English language proficiency (ELP) assessment systems for K-12 English learners (ELs) in the United States.The first paper articulated a high-level conceptualization of next-generation ELP assessment systems (Hauck, Wolf, & Mislevy,…

Diagnosis of a case of lipoid pneumonia by bronchoalveolar lavage.

PubMed

Spatafora, M; Bellia, V; Ferrara, G; Genova, G

1987-01-01

Exogenous lipoid pneumonia (ELP) was diagnosed by bronchoalveolar lavage (BAL) in a 57-year-old woman with a long history of using oily nose drops. Since clinical and roentgenological presentations are nonspecific, BAL, along with specific staining of recovered alveolar macrophages, represents a safe alternative to more invasive diagnostic procedures whenever a diagnosis of ELP is suspected.

Elastin-like-polypeptide based fusion proteins for osteogenic factor delivery in bone healing.

PubMed

McCarthy, Bryce; Yuan, Yuan; Koria, Piyush

2016-07-08

Modern treatments of bone injuries and diseases are becoming increasingly dependent on the usage of growth factors to stimulate bone growth. Bone morphogenetic protein-2 (BMP-2), a potent osteogenic inductive protein, exhibits promising results in treatment models, but recently has had its practical efficacy questioned due to the lack of local retention, ectopic bone formation, and potentially lethal inflammation. Where a new delivery technique of the BMP-2 is necessary, here we demonstrate the viability of an elastin-like peptide (ELP) fusion protein containing BMP-2 for delivery of the BMP-2. This fusion protein retains the performance characteristics of both the BMP-2 and ELP. The fusion protein was found to induce osteogenic differentiation of mesenchymal stem cells as evidenced by the production of alkaline phosphatase and extracellular calcium deposits in response to treatment by the fusion protein. Retention of the ELPs inverse phase transition property has allowed for expression of the fusion protein within a bacterial host (such as Escherichia coli) and easy and rapid purification using inverse transition cycling. The fusion protein formed self-aggregating nanoparticles at human-body temperature. The data collected suggests the viability of these fusion protein nanoparticles as a dosage-efficient and location-precise noncytotoxic delivery vehicle for BMP-2 in bone treatment. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1029-1037, 2016. © 2016 American Institute of Chemical Engineers.

The European Framework of Languages: A Piloting Sample of Cross-Curricular Strategy

ERIC Educational Resources Information Center

Mansilla, Paloma Ubeda; Riejos, Ana Maria Roldan

2007-01-01

This paper gives a short overview of the history of the Common European Framework of Reference (CEF) and European Language Portfolio (ELP) and explains their aims and functions. It then provides a summary of the ELP use in Europe and in Spain by showing a pilot study of its implementation carried out at the Schools of Civil Engineering and…

The International Language Portfolio: Appropriating a Tool for Learner Empowerment from the Council of Europe.

ERIC Educational Resources Information Center

Templar, Bill

2002-01-01

Foreign language educators need creative tools to empower students to make the languages they are learning truly their own. One such new instrument well worth inventive appropriation in Asia is the European Language Portfolio (ELP), launched by the Council of Europe in 2001. This report focuses on the three parts of ELP--a language passport,…

Next-Generation Summative English Language Proficiency Assessments for English Learners: Priorities for Policy and Research. Research Report. ETS RR-16-08

ERIC Educational Resources Information Center

Wolf, Mikyung Kim; Guzman-Orth, Danielle; Hauck, Maurice Cogan

2016-01-01

This paper is the third in a series concerning English language proficiency (ELP) assessments for K-12 English learners (ELs). The series, produced from Educational Testing Service (ETS), is intended to provide theory- and evidence-based principles and recommendations for improving next-generation ELP assessment systems, policies, and practices…

Characterization of Mixed Polypeptide Colloidal Particles by Light Scattering

NASA Astrophysics Data System (ADS)

Shuman, Hannah E.; Gaeckle, Grace K.; Gavin, John; Holland, Nolan B.; Streletzky, Kiril A.

2014-03-01

Temperature-dependent polymer surfactants have been developed by connecting three elastin-like polypeptide (ELP) chains to a charged protein domain (foldon), forming a three-armed star polymer. At low temperatures the polymer is soluble, while at higher temperatures it forms micelles. The behavior of mixtures of the three-armed star ELP (E20-Foldon) and H40-Linear ELP chains was analyzed under different salt and protein concentrations and various foldon to linear ELP ratio using Depolarized Dynamic Light Scattering. It was expected that under certain conditions the pure E20-Foldon would form spherical micelles, which upon adding the linear ELP would change in size and possibly shape. The pure E20-Foldon indeed formed largely spherical micelles with Rh of 10-20nm in solutions with 15-100mM salt and protein concentration between 10 μM and 100 μM. For the mixtures of 50 μM E20-Foldon and varying concentrations of H40-Linear in 25mM of salt, it was discovered that low and high H40-Linear concentration (4 μM and 50 μM) had only one transition. For the mixtures with of 10 and 25 μM of H40-Linear the two distinct transition temperatures were observed by spectrophotometry. The first transition corresponded to significantly elongated diffusive particles of apparent Rh of 30-50nm, while the second transition corresponded to slightly anisotropic diffusive particles with apparent Rh of about 20nm. At all H40-Linear concentrations studied, diffusive particles were seen above the second transition. Their radius and ability to depolarize light increased with the increase of H40-Linear concentration.

High-level production of human interleukin-10 fusions in tobacco cell suspension cultures

PubMed Central

Kaldis, Angelo; Ahmad, Adil; Reid, Alexandra; McGarvey, Brian; Brandle, Jim; Ma, Shengwu; Jevnikar, Anthony; Kohalmi, Susanne E; Menassa, Rima

2013-01-01

The production of pharmaceutical proteins in plants has made much progress in recent years with the development of transient expression systems, transplastomic technology and humanizing glycosylation patterns in plants. However, the first therapeutic proteins approved for administration to humans and animals were made in plant cell suspensions for reasons of containment, rapid scale-up and lack of toxic contaminants. In this study, we have investigated the production of human interleukin-10 (IL-10) in tobacco BY-2 cell suspension and evaluated the effect of an elastin-like polypeptide tag (ELP) and a green fluorescent protein (GFP) tag on IL-10 accumulation. We report the highest accumulation levels of hIL-10 obtained with any stable plant expression system using the ELP fusion strategy. Although IL-10-ELP has cytokine activity, its activity is reduced compared to unfused IL-10, likely caused by interference of ELP with folding of IL-10. Green fluorescent protein has no effect on IL-10 accumulation, but examining the trafficking of IL-10-GFP over the cell culture cycle revealed fluorescence in the vacuole during the stationary phase of the culture growth cycle. Analysis of isolated vacuoles indicated that GFP alone is found in vacuoles, while the full-size fusion remains in the whole-cell extract. This indicates that GFP is cleaved off prior to its trafficking to the vacuole. On the other hand, IL-10-GFP-ELP remains mostly in the ER and accumulates to high levels. Protein bodies were observed at the end of the culture cycle and are thought to arise as a consequence of high levels of accumulation in the ER. PMID:23297698

Using an e-Portfolio to Facilitate the Self-Assessment of Both Language and Intercultural Learning in Higher Education: A Case-Study Approach

ERIC Educational Resources Information Center

Kennedy, Fionnuala; Bruen, Jenny; Péchenart, Juliette

2012-01-01

The focus of this paper is twofold: first, on the development of an electronic version of a European Language Portfolio (ELP), known as the LOLIPOP (Language On-line Portfolio Project) ELP, and second on its integration into an undergraduate module on Intercultural Communication in an institute of higher education in Ireland. The paper begins by…

A Genetically Engineered Thermally Responsive Sustained Release Curcumin Depot to Treat Neuroinflammation

PubMed Central

Sinclair, S. Michael; Bhattacharyya, Jayanta; McDaniel, Jonathan R.; Gooden, David M.; Gopalaswamy, Ramesh; Chilkoti, Ashutosh; Setton, Lori A.

2014-01-01

Radiculopathy, a painful neuroinflammation that can accompany intervertebral disc herniation, is associated with locally increased levels of the pro-inflammatory cytokine tumor necrosis factor alpha (TNFα). Systemic administration of TNF antagonists for radiculopathy in the clinic has shown mixed results, and there is growing interest in the local delivery of anti-inflammatory drugs to treat this pathology as well as similar inflammatory events of peripheral nerve injury. Curcumin, a known antagonist of TNFα in multiple cell types and tissues, was chemically modified and conjugated to a thermally responsive elastin-like polypeptide (ELP) to create an injectable depot for sustained, local delivery of curcumin to treat neuroinflammation. ELPs are biopolymers capable of thermally-triggered in situ depot formation that have been successfully employed as drug carriers and biomaterials in several applications. ELP-curcumin conjugates were shown to display high drug loading, rapidly release curcumin in vitro via degradable carbamate bonds, and retain in vitro bioactivity against TNFα-induced cytotoxicity and monocyte activation with IC50 only two-fold higher than curcumin. When injected proximal to the sciatic nerve in mice via intramuscular (i.m.) injection, ELP-curcumin conjugates underwent a thermally triggered soluble-insoluble phase transition, leading to in situ formation of a depot that released curcumin over 4 days post-injection and decreased plasma AUC 7-fold. PMID:23830979

English Language Proficiency and Content Assessment Performance: A Comparison of English Learners and Native English Speakers Achievement

ERIC Educational Resources Information Center

Miley, Suzi Keller; Farmer, Aarek

2017-01-01

As a result of the accountability requirements established in Title III of the Elementary and Secondary Educational Act (ESEA) legislation, English Learners (ELs) are expected to make progress in both content area academic achievement and English Language Proficiency (ELP). In Tennessee ELs progress is measured by administering WIDA-Access to…

Exogenous Lipoid Pneumonia in Laryngectomy Patients: Radiological Findings.

PubMed

García Latorre, Raquel; Rodríguez Díaz, Ricardo; Barrios Barreto, Deisy; Ayala Carbonero, Ana; García Gómez-Muriel, María Isabel; Gorospe Sarasúa, Luis

2015-07-01

Exogenous lipoid pneumonia (ELP) is a rare (incidence 1.0%-2.5%), often under-diagnosed disease, caused by the aspiration and accumulation of exogenous lipids within the pulmonary alveoli. Various cases have been described due to inhalation of lubricants via the nasal passages and oropharynx, aspiration of mineral oils in laxatives in patients with eating disorders, application of lip gloss, occupational exposure to liquid paraffin or mineral oils ("fire-eaters", industrial use in washing of machinery, automobile workshops, plastic paints, etc.) and application of Vaseline during the insertion of nasogastric tubes and in the care of tracheotomy patients. ELP usually presents radiologically as areas of low-attenuation peribronchial consolidation and ground glass opacities, with a predominantly bibasal distribution. We present 5 cases of long-standing laryngectomy patients diagnosed with ELP who admitted using Vaseline in their tracheal stoma care. Copyright © 2014 SEPAR. Published by Elsevier Espana. All rights reserved.

Enterocolic lymphocytic phlebitis of the cecal pole and appendix vermiformis with increase of IgG4-positive plasma cells.

PubMed

Comtesse, Sarah; Friemel, Juliane; Fankhauser, René; Weber, Achim

2014-01-01

Here we describe the clinicopathological course of a 20-year-old female patient with enterocolic lymphocytic phlebitis (ELP) of the appendix vermiformis and cecal pole with increase of IgG4-positive plasma cells. The patient presented with acute abdomen, suspicious of acute appendicitis. Diagnostic laparoscopy showed tumefaction of the cecal pole and appendix vermiformis. Histologic examination revealed mural thickening and a dense lymphoplasmocytic, partly obliterative infiltrate of the veins with sparing of the arteries, diagnostic of ELP. In addition, we found an elevated number of IgG4-positive plasma cells blended in with the lymphocytes. The IgG4-to-IgG ratio accounted for >40 %. This case meets the histopathological criteria requested for IgG4-related disease (IgG4-RD) and thus opens the possibility that ELP might be part of the IgG4-RD spectrum.

Targeting prostate cancer cells with hybrid elastin-like polypeptide/liposome nanoparticles

PubMed Central

Zhang, Wei; Song, Yunmei; Eldi, Preethi; Guo, Xiuli; Hayball, John D; Garg, Sanjay; Albrecht, Hugo

2018-01-01

Prostate cancer cells frequently overexpress the gastrin-releasing peptide receptor, and various strategies have been applied in preclinical settings to target this receptor for the specific delivery of anticancer compounds. Recently, elastin-like polypeptide (ELP)-based self-assembling micelles with tethered GRP on the surface have been suggested to actively target prostate cancer cells. Poorly soluble chemotherapeutics such as docetaxel (DTX) can be loaded into the hydrophobic cores of ELP micelles, but only limited drug retention times have been achieved. Herein, we report the generation of hybrid ELP/liposome nanoparticles which self-assembled rapidly in response to temperature change, encapsulated DTX at high concentrations with slow release, displayed the GRP ligand on the surface, and specifically bound to GRP receptor expressing PC-3 cells as demonstrated by flow cytometry. This novel type of drug nanocarrier was successfully used to reduce cell viability of prostate cancer cells in vitro through the specific delivery of DTX. PMID:29391790

Bio-inspired synthesis of hybrid silica nanoparticles templated from elastin-like polypeptide micelles

NASA Astrophysics Data System (ADS)

Han, Wei; MacEwan, Sarah R.; Chilkoti, Ashutosh; López, Gabriel P.

2015-07-01

The programmed self-assembly of block copolymers into higher order nanoscale structures offers many attractive attributes for the development of new nanomaterials for numerous applications including drug delivery and biosensing. The incorporation of biomimetic silaffin peptides in these block copolymers enables the formation of hybrid organic-inorganic materials, which can potentially enhance the utility and stability of self-assembled nanostructures. We demonstrate the design, synthesis and characterization of amphiphilic elastin-like polypeptide (ELP) diblock copolymers that undergo temperature-triggered self-assembly into well-defined spherical micelles. Genetically encoded incorporation of the silaffin R5 peptide at the hydrophilic terminus of the diblock ELP leads to presentation of the silaffin R5 peptide on the coronae of the micelles, which results in localized condensation of silica and the formation of near-monodisperse, discrete, sub-100 nm diameter hybrid ELP-silica particles. This synthesis method, can be carried out under mild reaction conditions suitable for bioactive materials, and will serve as the basis for the development and application of functional nanomaterials. Beyond silicification, the general strategies described herein may also be adapted for the synthesis of other biohybrid nanomaterials as well.The programmed self-assembly of block copolymers into higher order nanoscale structures offers many attractive attributes for the development of new nanomaterials for numerous applications including drug delivery and biosensing. The incorporation of biomimetic silaffin peptides in these block copolymers enables the formation of hybrid organic-inorganic materials, which can potentially enhance the utility and stability of self-assembled nanostructures. We demonstrate the design, synthesis and characterization of amphiphilic elastin-like polypeptide (ELP) diblock copolymers that undergo temperature-triggered self-assembly into well-defined spherical micelles. Genetically encoded incorporation of the silaffin R5 peptide at the hydrophilic terminus of the diblock ELP leads to presentation of the silaffin R5 peptide on the coronae of the micelles, which results in localized condensation of silica and the formation of near-monodisperse, discrete, sub-100 nm diameter hybrid ELP-silica particles. This synthesis method, can be carried out under mild reaction conditions suitable for bioactive materials, and will serve as the basis for the development and application of functional nanomaterials. Beyond silicification, the general strategies described herein may also be adapted for the synthesis of other biohybrid nanomaterials as well. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr01407g

Development of Intra-Articular Drug Delivery to Alter Progression of Arthritis Following Joint Injury

DTIC Science & Technology

2012-04-01

2012Revised AnnualApril 2012 DESIGN: A novel injectable and in situ forming drug depot based on thermally-responsive elastin -like polypeptide (ELP) will... Elastin -like polypeptide, Drug Depot – technology allowing sustained release of biologically active agent , Active agents used include IL1Ra...The abstract has been removed and an appendix has been included. In brief this protocol explores the use of elastin like polypeptide (ELP) as a

Treatment of the Cornea Using Transcytotic Delivery into the Tear Film

DTIC Science & Technology

2015-12-01

early April 2013; however, we continue to produce additional material as required (Fig. 4). In milestone 3 we initiated in vivo evaluation of...SI, the minimum and maximum values are 12.4 and 34.4. 8 Aim 2) characterize Lacritin-ELP fusion proteins and evaluate as therapeutic agents...0.0001). Data were analyzed by a two-way ANOVA followed by Tukey’s multiple comparisons test (n=9). 10 Having evaluated the lacritin-ELP fusion in

Fostering Engagement with the "Common European Framework of Reference for Languages" and the European Language Portfolio: Learning from Good Practice in University Language Centres

ERIC Educational Resources Information Center

Carson, Lorna

2016-01-01

This article begins with a brief overview of the aims and activities of the CercleS CEFR/ELP Focus Group. It goes on to report on some of the outcomes of the 2015 CercleS CEFR/ELP seminar hosted by the Centre for Language and Communication Studies at Trinity College Dublin. The five examples presented during the seminar's plenary sessions are…

In vitro characterization of a collagen scaffold enzymatically cross-linked with a tailored elastin-like polymer.

PubMed

Garcia, Yolanda; Hemantkumar, Naik; Collighan, Russell; Griffin, Martin; Rodriguez-Cabello, Jose Carlos; Pandit, Abhay

2009-04-01

Collagen, the main structural component of the extracellular matrix (ECM), provides tensile stiffness to different structures and organs against rupture. However, collagen tissue-engineered implants are hereto still lacking in mechanical strength. Attempts to create stiffer scaffolds have resulted in increased brittleness of the material, reducing the versatility of the original component. The hypothesis behind this research is that the introduction of an elastic element in the scaffold will enhance the mechanical properties of the collagen-based scaffolds, as elastin does in the ECM to prevent irreversible deformation. In this study, an elastin-like polymer (ELP) designed and synthesized using recombinant DNA methodology is used with the view to providing increased proteolytic resistance and increased functionality to the scaffolds by carrying specific sequences for microbial transglutaminase cross-linking, endothelial cell adhesion, and drug delivery. Evaluation of the effects that cross-linking ELP-collagen has on the physicochemical properties of the scaffold such as porosity, presence of cross-linking, thermal behavior, and mechanical strength demonstrated that the introduction of enzymatically resistant covalent bonds between collagen and ELP increases the mechanical strength of the scaffolds in a dose-dependent manner without significantly affecting the porosity or thermal properties of the original scaffold. Importantly, the scaffolds also showed selective behavior, in a dose (ELP)-dependent manner toward human umbilical vein endothelial cells and smooth muscle cells when compared to fibroblasts.

National Evaluation of Title III Implementation Supplemental Report: Exploring Approaches to Setting English Language Proficiency Performance Criteria and Monitoring English Learner Progress

ERIC Educational Resources Information Center

Cook, Gary; Linquanti, Robert; Chinen, Marjorie; Jung, Hyekyung

2012-01-01

The "Elementary and Secondary Education Act" ("ESEA"), as amended by the "No Child Left Behind Act of 2001" inaugurated important changes in assessment and accountability for English Learner (EL) students. Specifically, Title III of the law required states to develop or adopt English-language proficiency (ELP)…

Modification of tRNALys UUU by Elongator Is Essential for Efficient Translation of Stress mRNAs

PubMed Central

Sansó, Miriam; Buhne, Karin; Carmona, Mercè; Paulo, Esther; Hermand, Damien; Rodríguez-Gabriel, Miguel; Ayté, José; Leidel, Sebastian; Hidalgo, Elena

2013-01-01

The Elongator complex, including the histone acetyl transferase Sin3/Elp3, was isolated as an RNA polymerase II-interacting complex, and cells deficient in Elongator subunits display transcriptional defects. However, it has also been shown that Elongator mediates the modification of some tRNAs, modulating translation efficiency. We show here that the fission yeast Sin3/Elp3 is important for oxidative stress survival. The stress transcriptional program, governed by the Sty1-Atf1-Pcr1 pathway, is affected in mutant cells, but not severely. On the contrary, cells lacking Sin3/Elp3 cannot modify the uridine wobble nucleoside of certain tRNAs, and other tRNA modifying activities such as Ctu1-Ctu2 are also essential for normal tolerance to H2O2. In particular, a plasmid over-expressing the tRNALys UUU complements the stress-related phenotypes of Sin3/Elp3 mutant cells. We have determined that the main H2O2-dependent genes, including those coding for the transcription factors Atf1 and Pcr1, are highly expressed mRNAs containing a biased number of lysine-coding codons AAA versus AAG. Thus, their mRNAs are poorly translated after stress in cells lacking Sin3/Elp3 or Ctu2, whereas a mutated atf1 transcript with AAA-to-AAG lysine codons is efficiently translated in all strain backgrounds. Our study demonstrates that the lack of a functional Elongator complex results in stress phenotypes due to its contribution to tRNA modification and subsequent translation inefficiency of certain stress-induced, highly expressed mRNAs. These results suggest that the transcriptional defects of these strain backgrounds may be a secondary consequence of the deficient expression of a transcription factor, Atf1-Pcr1, and other components of the transcriptional machinery. PMID:23874237

Modification of tRNA(Lys) UUU by elongator is essential for efficient translation of stress mRNAs.

PubMed

Fernández-Vázquez, Jorge; Vargas-Pérez, Itzel; Sansó, Miriam; Buhne, Karin; Carmona, Mercè; Paulo, Esther; Hermand, Damien; Rodríguez-Gabriel, Miguel; Ayté, José; Leidel, Sebastian; Hidalgo, Elena

2013-01-01

The Elongator complex, including the histone acetyl transferase Sin3/Elp3, was isolated as an RNA polymerase II-interacting complex, and cells deficient in Elongator subunits display transcriptional defects. However, it has also been shown that Elongator mediates the modification of some tRNAs, modulating translation efficiency. We show here that the fission yeast Sin3/Elp3 is important for oxidative stress survival. The stress transcriptional program, governed by the Sty1-Atf1-Pcr1 pathway, is affected in mutant cells, but not severely. On the contrary, cells lacking Sin3/Elp3 cannot modify the uridine wobble nucleoside of certain tRNAs, and other tRNA modifying activities such as Ctu1-Ctu2 are also essential for normal tolerance to H2O2. In particular, a plasmid over-expressing the tRNA(Lys) UUU complements the stress-related phenotypes of Sin3/Elp3 mutant cells. We have determined that the main H2O2-dependent genes, including those coding for the transcription factors Atf1 and Pcr1, are highly expressed mRNAs containing a biased number of lysine-coding codons AAA versus AAG. Thus, their mRNAs are poorly translated after stress in cells lacking Sin3/Elp3 or Ctu2, whereas a mutated atf1 transcript with AAA-to-AAG lysine codons is efficiently translated in all strain backgrounds. Our study demonstrates that the lack of a functional Elongator complex results in stress phenotypes due to its contribution to tRNA modification and subsequent translation inefficiency of certain stress-induced, highly expressed mRNAs. These results suggest that the transcriptional defects of these strain backgrounds may be a secondary consequence of the deficient expression of a transcription factor, Atf1-Pcr1, and other components of the transcriptional machinery.

The Emergency Landing Planner Experiment

NASA Technical Reports Server (NTRS)

Meuleau, Nocolas F.; Neukom, Christian; Plaunt, Christian John; Smith, David E.; Smith, Tristan B.

2011-01-01

In previous work, we described an Emergency Landing Planner (ELP) designed to assist pilots in choosing the best emergency landing site when damage or failures occur in an aircraft. In this paper, we briefly describe the system, but focus on the integration of this system into the cockpit of a 6 DOF full-motion simulator and a study designed to evaluate the ELP. We discuss the results of this study, the lessons learned, and some of the issues involved in advancing this work further.

Oligomerization triggered by foldon: a simple method to enhance the catalytic efficiency of lichenase and xylanase.

PubMed

Wang, Xinzhe; Ge, Huihua; Zhang, Dandan; Wu, Shuyu; Zhang, Guangya

2017-07-03

Effective and simple methods that lead to higher enzymatic efficiencies are highly sough. Here we proposed a foldon-triggered trimerization of the target enzymes with significantly improved catalytic performances by fusing a foldon domain at the C-terminus of the enzymes via elastin-like polypeptides (ELPs). The foldon domain comprises 27 residues and can forms trimers with high stability. Lichenase and xylanase can hydrolyze lichenan and xylan to produce value added products and biofuels, and they have great potentials as biotechnological tools in various industrial applications. We took them as the examples and compared the kinetic parameters of the engineered trimeric enzymes to those of the monomeric and wild type ones. When compared with the monomeric ones, the catalytic efficiency (k cat /K m ) of the trimeric lichenase and xylanase increased 4.2- and 3.9- fold. The catalytic constant (k cat ) of the trimeric lichenase and xylanase increased 1.8- fold and 5.0- fold than their corresponding wild-type counterparts. Also, the specific activities of trimeric lichenase and xylanase increased by 149% and 94% than those of the monomeric ones. Besides, the recovery of the lichenase and xylanase activities increased by 12.4% and 6.1% during the purification process using ELPs as the non-chromatographic tag. The possible reason is the foldon domain can reduce the transition temperature of the ELPs. The trimeric lichenase and xylanase induced by foldon have advantages in the catalytic performances. Besides, they were easier to purify with increased purification fold and decreased the loss of activities compared to their corresponding monomeric ones. Trimerizing of the target enzymes triggered by the foldon domain could improve their activities and facilitate the purification, which represents a simple and effective enzyme-engineering tool. It should have exciting potentials both in industrial and laboratory scales.

Estimation of Mangrove Net Primary Production and Carbon Sequestration service using Light Use Efficiency model in the Sunderban Biosphere region, India

NASA Astrophysics Data System (ADS)

Sannigrahi, Srikanta; Sen, Somnath; Paul, Saikat

2016-04-01

Net Primary Production (NPP) of mangrove ecosystem and its capacity to sequester carbon from the atmosphere may be used to quantify the regulatory ecosystem services. Three major group of parameters has been set up as BioClimatic Parameters (BCP): (Photosynthetically Active Radiation (PAR), Absorbed PAR (APAR), Fraction of PAR (FPAR), Photochemical Reflectance Index (PRI), Light Use Efficiency (LUE)), BioPhysical Parameters (BPP) :(Normalize Difference Vegetation Index (NDVI), scaled NDVI, Enhanced Vegetation Index (EVI), scaled EVI, Optimised and Modified Soil Adjusted Vegetation Index (OSAVI, MSAVI), Leaf Area Index (LAI)), and Environmental Limiting Parameters (ELP) (Temperature Stress (TS), Land Surface Water Index (LSWI), Normalize Soil Water Index (NSWI), Water Stress Scalar (WS), Inversed WS (iWS) Land Surface Temperature (LST), scaled LST, Vapor Pressure Deficit (VPD), scaled VPD, and Soil Water Deficit Index (SWDI)). Several LUE models namely Carnegie Ames Stanford Approach (CASA), Eddy Covariance - LUE (EC-LUE), Global Production Efficiency Model (GloPEM), Vegetation Photosynthesis Model (VPM), MOD NPP model, Temperature and Greenness Model (TG), Greenness and Radiation model (GR) and MOD17 was adopted in this study to assess the spatiotemporal nature of carbon fluxes. Above and Below Ground Biomass (AGB & BGB) was calculated using field based estimation of OSAVI and NDVI. Microclimatic zonation has been set up to assess the impact of coastal climate on environmental limiting factors. MODerate Resolution Imaging Spectroradiometer (MODIS) based yearly Gross Primary Production (GPP) and NPP product MOD17 was also tested with LUE based results with standard model validation statistics: Root Mean Square of Error (RMSE), Mean Absolute Error (MEA), Bias, Coefficient of Variation (CV) and Coefficient of Determination (R2). The performance of CASA NPP was tested with the ground based NPP with R2 = 0.89 RMSE = 3.28 P = 0.01. Among the all adopted models, EC-LUE and VPM models has explained the maximum variances (>80%) in comparison to the other model. Study result has also showed that the BPP has explained the maximum model variances (>93%) followed by BCP (>65%) and ELP (>50%). Scaled WS, iWS, LST, VPD, NDVI was performed better in a minimum ELP condition whereas surface moisture and wetness was highly correlated with the AGB and NPP (R2 = 0.86 RMSE = 1.83). During this study period (2000-2013), it was found that there was a significantly declining trend (R2 = 0.32 P = 0.05) of annual NPP and the maximum decrease was found in the eastern part where built-up area was mainly accounted for reduction of NPP. BCP are explained higher variances (>80%) in the optimum climatic condition exist along the coastal stretches in comparison to the landward extent (>45%).

High-yield recombinant expression and purification of marginally soluble, short elastin-like polypeptides.

PubMed

Bahniuk, Markian S; Alshememry, Abdullah K; Unsworth, Larry D

2016-12-01

The protocol described here is designed as an extension of existing techniques for creating elastin-like polypeptides. It allows for the expression and purification of elastin-like polypeptide (ELP) constructs that are poorly expressed or have very low transition temperatures. DNA concatemerization has been modified to reduce issues caused by methylation sensitivity and inefficient cloning. Linearization of the modified expression vector has been altered to greatly increase cleavage efficiency. The purification regimen is based upon using denaturing metal affinity chromatography to fully solubilize and, if necessary, pre-concentrate the target peptide before purification by inverse temperature cycling (ITC). This protocol has been used to express multiple leucine-containing elastin-like polypeptides, with final yields of 250-660 mg per liter of cells, depending on the specific construct. This was considerably greater than previously reported yields for similar ELPs. Due to the relative hydrophobicity of the tested constructs, even compared with commonly employed ELPs, conventional methods would not have been able to be purify these peptides.

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

Ghosh, Koushik; Balog, Eva Rose M.; Sista, Prakash

We report a method for creating hybrid organic-inorganic “nanoflowers” using calcium or copper ions as the inorganic component and a recombinantly expressed elastin-like polypeptide (ELP) as the organic component. Polypeptides provide binding sites for the dynamic coordination with metal ions, and then such noncovalent complexes become nucleation sites for primary crystals of metal phosphates. We have shown that the interaction between the stimuli-responsive ELP and Ca{sup 2+} or Cu{sup 2+}, in the presence of phosphate, leads to the growth of micrometer-sized particles featuring nanoscale patterns shaped like flower petals. The morphology of these flower-like composite structures is dependent upon themore » temperature of growth and has been characterized by scanning electron microscopy. The composition of nanoflowers has also been analyzed by energy-dispersive X-ray spectroscopy, X-ray photoelectron spectroscopy, and X-ray diffraction. The temperature-dependent morphologies of these hybrid nanostructures, which arise from the controllable phase transition of ELPs, hold potential for morphological control of biomaterials in emerging applications such as tissue engineering and biocatalysis.« less

Loss of Elp3 Impairs the Acetylation and Distribution of Connexin-43 in the Developing Cerebral Cortex

PubMed Central

Laguesse, Sophie; Close, Pierre; Van Hees, Laura; Chariot, Alain; Malgrange, Brigitte; Nguyen, Laurent

2017-01-01

The Elongator complex is required for proper development of the cerebral cortex. Interfering with its activity in vivo delays the migration of postmitotic projection neurons, at least through a defective α-tubulin acetylation. However, this complex is already expressed by cortical progenitors where it may regulate the early steps of migration by targeting additional proteins. Here we report that connexin-43 (Cx43), which is strongly expressed by cortical progenitors and whose depletion impairs projection neuron migration, requires Elongator expression for its proper acetylation. Indeed, we show that Cx43 acetylation is reduced in the cortex of Elp3cKO embryos, as well as in a neuroblastoma cell line depleted of Elp1 expression, suggesting that Cx43 acetylation requires Elongator in different cellular contexts. Moreover, we show that histones deacetylase 6 (HDAC6) is a deacetylase of Cx43. Finally, we report that acetylation of Cx43 regulates its membrane distribution in apical progenitors of the cerebral cortex. PMID:28507509

Solvent Properties of Water in Aqueous Solutions of Elastin-Like Polypeptide

PubMed Central

Ferreira, Luisa A.; Cole, James T.; Reichardt, Christian; Holland, Nolan B.; Uversky, Vladimir N.; Zaslavsky, Boris Y.

2015-01-01

The phase-transition temperatures of an elastin-like polypeptide (ELP) with the (GVGVP)40 sequence and solvent dipolarity/polarizability, hydrogen-bond donor acidity, and hydrogen-bond acceptor basicity in its aqueous solutions were quantified in the absence and presence of different salts (Na2SO4, NaCl, NaClO4, and NaSCN) and various osmolytes (sucrose, sorbitol, trehalose, and trimethylamine N-oxide (TMAO)). All osmolytes decreased the ELP phase-transition temperature, whereas NaCl and Na2SO4 decreased, and NaSCN and NaClO4 increased it. The determined phase-transition temperatures may be described as a linear combination of the solvent’s dipolarity/polarizability and hydrogen-bond donor acidity. The linear relationship established for the phase-transition temperature in the presence of salts differs quantitatively from that in the presence of osmolytes, in agreement with different (direct and indirect) mechanisms of the influence of salts and osmolytes on the ELP phase-transition temperature. PMID:26075870

Enterocolic lymphocytic phlebitis as a newly recognized manifestation of IgG4-related disease.

PubMed

Laco, Jan; Örhalmi, Július; Bártová, Jolana; Zimandlová, Dana

2015-04-01

Herein we present a case of a 65-year-old woman with enterocolic lymphocytic phlebitis (ELP) who presented with anemic syndrome and in whom severe stenosis of the right flexure of large bowel was detected. The microscopic examination revealed fibrosis of the submucosa and lymphoplasmacytic phlebitis of small veins and venules, whereas arteries were spared. There were 110 IgG4-positive and 160 IgG-positive plasma cells in 1 high-power field, respectively, with corresponding IgG4/IgG ratio of 0.69. The IgG4 serum level was 2.42 g/L. According to the currently proposed criteria, this ELP case is the first that may be diagnosed as definite IgG4-related disease (IgG4-RD). Although based on the sole case description, taken together with a recent review and a case report, we presume that a subset of ELPs is a manifestation of IgG4-RD. © The Author(s) 2014.

Electron Pair Repulsion Responsible for the Peculiar Edge Effects and Surface Chemistry of Black Phosphorus.

PubMed

Kong, Xiang-Peng; Shen, Xiaomei; Jang, Joonkyung; Gao, Xingfa

2018-03-01

The electronic and optical properties of black phosphorus (black-P) are significantly modulated by fabricating the edges of this two-dimensional material. Electron lone pairs (ELPs) are ubiquitous in black-P, but their role in creating the edge effects of black-P is poorly understood. Using first-principle calculations, we report ELPs of black-P experience severe Coulomb repulsion and play a central role in creating the edge effects of black-P. We discover the outermost P atoms of the zigzag edges of black-PQDs are free of the Coulomb repulsion, but the P atoms of the armchair edges do experience the Coulomb repulsion. The Coulomb repulsion serves as a new chemical driving force to make electron donor-acceptor bonds with chemical groups bearing vacant orbitals. Our results provide insights into the mechanism responsible for the peculiar edge effects of black-P and highlight the opportunity to use the ELPs of black-P for their damage-free surface functionalization.

Toughening of Thermoresponsive Arrested Networks of Elastin-Like Polypeptides To Engineer Cytocompatible Tissue Scaffolds.

PubMed

Glassman, Matthew J; Avery, Reginald K; Khademhosseini, Ali; Olsen, Bradley D

2016-02-08

Formulation of tissue engineering or regenerative scaffolds from simple bioactive polymers with tunable structure and mechanics is crucial for the regeneration of complex tissues, and hydrogels from recombinant proteins, such as elastin-like polypeptides (ELPs), are promising platforms to support these applications. The arrested phase separation of ELPs has been shown to yield remarkably stiff, biocontinuous, nanostructured networks, but these gels are limited in applications by their relatively brittle nature. Here, a gel-forming ELP is chain-extended by telechelic oxidative coupling, forming extensible, tough hydrogels. Small angle scattering indicates that the chain-extended polypeptides form a fractal network of nanoscale aggregates over a broad concentration range, accessing moduli ranging from 5 kPa to over 1 MPa over a concentration range of 5-30 wt %. These networks exhibited excellent erosion resistance and allowed for the diffusion and release of encapsulated particles consistent with a bicontinuous, porous structure with a broad distribution of pore sizes. Biofunctionalized, toughened networks were found to maintain the viability of human mesenchymal stem cells (hMSCs) in 2D, demonstrating signs of osteogenesis even in cell media without osteogenic molecules. Furthermore, chondrocytes could be readily mixed into these gels via thermoresponsive assembly and remained viable in extended culture. These studies demonstrate the ability to engineer ELP-based arrested physical networks on the molecular level to form reinforced, cytocompatible hydrogel matrices, supporting the promise of these new materials as candidates for the engineering and regeneration of stiff tissues.

Protein Bodies in Leaves Exchange Contents through the Endoplasmic Reticulum

DOE PAGES

Saberianfar, Reza; Sattarzadeh, Amirali; Joensuu, Jussi J.; ...

2016-05-23

Protein bodies (PBs) are organelles found in seeds whose main function is the storage of proteins that are used during germination for sustaining growth. PBs can also be induced to form in leaves when foreign proteins are produced at high levels in the endoplasmic reticulum (ER) and when fused to one of three tags: Zera®, elastin-like polypeptides (ELP), or hydrophobin-I (HFBI). Here in this study, we investigate the differences between ELP, HFBI and Zera PB formation, packing, and communication. Our results confirm the ER origin of all three fusion-tag-induced PBs. We show that secretory pathway proteins can be sequestered intomore » all types of PBs but with different patterns, and that different fusion tags can target a specific protein to different PBs. Zera PBs are mobile and dependent on actomyosin motility similar to ELP and HFBI PBs. We show in vivo trafficking of proteins between PBs using GFP photoconversion. We also show that protein trafficking between ELP or HFBI PBs is faster and proteins travel further when compared to Zera PBs. Our results indicate that fusion-tag-induced PBs do not represent terminally stored cytosolic organelles, but that they form in, and remain part of the ER, and dynamically communicate with each other via the ER. We hypothesize that the previously documented PB mobility along the actin cytoskeleton is associated with ER movement rather than independent streaming of detached organelles.« less

Subzero water permeability parameters and optimal freezing rates for sperm cells of the southern platyfish, Xiphophorus maculatus.

PubMed

Pinisetty, D; Huang, C; Dong, Q; Tiersch, T R; Devireddy, R V

2005-06-01

This study reports the subzero water transport characteristics (and empirically determined optimal rates for freezing) of sperm cells of live-bearing fishes of the genus Xiphophorus, specifically those of the southern platyfish Xiphophorus maculatus. These fishes are valuable models for biomedical research and are commercially raised as ornamental fish for use in aquariums. Water transport during freezing of X. maculatus sperm cell suspensions was obtained using a shape-independent differential scanning calorimeter technique in the presence of extracellular ice at a cooling rate of 20 degrees C/min in three different media: (1) Hanks' balanced salt solution (HBSS) without cryoprotective agents (CPAs); (2) HBSS with 14% (v/v) glycerol, and (3) HBSS with 10% (v/v) dimethyl sulfoxide (DMSO). The sperm cell was modeled as a cylinder with a length of 52.35 microm and a diameter of 0.66 microm with an osmotically inactive cell volume (Vb) of 0.6 V0, where V0 is the isotonic or initial cell volume. This translates to a surface area, SA to initial water volume, WV ratio of 15.15 microm(-1). By fitting a model of water transport to the experimentally determined volumetric shrinkage data, the best fit membrane permeability parameters (reference membrane permeability to water at 0 degrees C, Lpg or Lpg [cpa] and the activation energy, E(Lp) or E(Lp) [cpa]) were found to range from: Lpg or Lpg [cpa] = 0.0053-0.0093 microm/minatm; E(Lp) or E(Lp) [cpa] = 9.79-29.00 kcal/mol. By incorporating these membrane permeability parameters in a recently developed generic optimal cooling rate equation (optimal cooling rate, [Formula: see text] where the units of B(opt) are degrees C/min, E(Lp) or E(Lp) [cpa] are kcal/mol, L(pg) or L(pg) [cpa] are microm/minatm and SA/WV are microm(-1)), we determined the optimal rates of freezing X. maculatus sperm cells to be 28 degrees C/min (in HBSS), 47 degrees C/min (in HBSS+14% glycerol) and 36 degrees C/min (in HBSS+10% DMSO). Preliminary empirical experiments suggest that the optimal rate of freezing X. maculatus sperm in the presence of 14% glycerol to be approximately 25 degrees C/min. Possible reasons for the observed discrepancy between the theoretically predicted and experimentally determined optimal rates of freezing X. maculatus sperm cells are discussed.

A fast BK-type KCa current acts as a postsynaptic modulator of temporal selectivity for communication signals.

PubMed

Kohashi, Tsunehiko; Carlson, Bruce A

2014-01-01

Temporal patterns of spiking often convey behaviorally relevant information. Various synaptic mechanisms and intrinsic membrane properties can influence neuronal selectivity to temporal patterns of input. However, little is known about how synaptic mechanisms and intrinsic properties together determine the temporal selectivity of neuronal output. We tackled this question by recording from midbrain electrosensory neurons in mormyrid fish, in which the processing of temporal intervals between communication signals can be studied in a reduced in vitro preparation. Mormyrids communicate by varying interpulse intervals (IPIs) between electric pulses. Within the midbrain posterior exterolateral nucleus (ELp), the temporal patterns of afferent spike trains are filtered to establish single-neuron IPI tuning. We performed whole-cell recording from ELp neurons in a whole-brain preparation and examined the relationship between intrinsic excitability and IPI tuning. We found that spike frequency adaptation of ELp neurons was highly variable. Postsynaptic potentials (PSPs) of strongly adapting (phasic) neurons were more sharply tuned to IPIs than weakly adapting (tonic) neurons. Further, the synaptic filtering of IPIs by tonic neurons was more faithfully converted into variation in spiking output, particularly at short IPIs. Pharmacological manipulation under current- and voltage-clamp revealed that tonic firing is mediated by a fast, large-conductance Ca(2+)-activated K(+) (KCa) current (BK) that speeds up action potential repolarization. These results suggest that BK currents can shape the temporal filtering of sensory inputs by modifying both synaptic responses and PSP-to-spike conversion. Slow SK-type KCa currents have previously been implicated in temporal processing. Thus, both fast and slow KCa currents can fine-tune temporal selectivity.

The influence of surfactant HLB and oil/surfactant ratio on the formation and properties of self-emulsifying pellets and microemulsion reconstitution.

PubMed

Matsaridou, Irini; Barmpalexis, Panagiotis; Salis, Andrea; Nikolakakis, Ioannis

2012-12-01

Self-emulsifying oil/surfactant mixtures can be incorporated into pellets that have the advantages of the oral administration of both microemulsions and a multiple-unit dosage form. The purpose of this work was to study the effects of surfactant hydrophilic-lipophilic balance (HLB) and oil/surfactant ratio on the formation and properties of self-emulsifying microcrystalline cellulose (MCC) pellets and microemulsion reconstitution. Triglycerides (C(8)-C(10)) was the oil and Cremophor ELP and RH grades and Solutol the surfactants. Pellets were prepared by extrusion/spheronization using microemulsions with fixed oil/surfactant content but with different water proportions to optimize size and shape parameters. Microemulsion reconstitution from pellets suspended in water was evaluated by turbidimetry and light scattering size analysis, and H-bonding interactions of surfactant with MCC from FT-IR spectra. It was found that water requirements for pelletization increased linearly with increasing HLB. Crushing load decreased and deformability increased with increasing oil/surfactant ratio. Incorporation of higher HLB surfactants enhanced H-bonding and resulted in faster and more extensive disintegration of MCC as fibrils. Reconstitution was greater at high oil/surfactant ratios and the droplet size of the reconstituted microemulsions was similar to that in the wetting microemulsions. The less hydrophilic ELP with a double bond in the fatty acid showed weaker H-bonding and greater microemulsion reconstitution. Purified ELP gave greater reconstitution than the unpurified grade. Thus, the work demonstrates that the choice of type and quantity of the surfactant used in the formulation of microemulsions containing pellets has an important influence on their production and performance.

Temporal-pattern recognition by single neurons in a sensory pathway devoted to social communication behavior

PubMed Central

Carlson, Bruce A.

2010-01-01

Sensory systems often encode stimulus information into the temporal pattern of action potential activity. However, little is known about how the information contained within these patterns is extracted by postsynaptic neurons. Similar to temporal coding by sensory neurons, social information in mormyrid fish is encoded into the temporal patterning of an electric organ discharge (EOD). In the current study, sensitivity to temporal patterns of electrosensory stimuli was found to arise within the midbrain posterior exterolateral nucleus (ELp). Whole-cell patch recordings from ELp neurons in vivo revealed three patterns of interpulse interval (IPI) tuning: low-pass neurons tuned to long intervals, high-pass neurons tuned to short intervals and band-pass neurons tuned to intermediate intervals. Many neurons within each class also responded preferentially to either increasing or decreasing IPIs. Playback of electric signaling patterns recorded from freely behaving fish revealed that the IPI and direction tuning of ELp neurons resulted in selective responses to particular social communication displays characterized by distinct IPI patterns. The postsynaptic potential responses of many neurons indicated a combination of excitatory and inhibitory synaptic input, and the IPI tuning of ELp neurons was directly related to rate-dependent changes in the direction and amplitude of postsynaptic potentials. These results suggest that differences in the dynamics of short-term synaptic plasticity in excitatory and inhibitory pathways may tune central sensory neurons to particular temporal patterns of presynaptic activity. This may represent a general mechanism for the processing of behaviorally-relevant stimulus information encoded into temporal patterns of activity by sensory neurons. PMID:19641105

Temporal-pattern recognition by single neurons in a sensory pathway devoted to social communication behavior.

PubMed

Carlson, Bruce A

2009-07-29

Sensory systems often encode stimulus information into the temporal pattern of action potential activity. However, little is known about how the information contained within these patterns is extracted by postsynaptic neurons. Similar to temporal coding by sensory neurons, social information in mormyrid fish is encoded into the temporal patterning of an electric organ discharge. In the current study, sensitivity to temporal patterns of electrosensory stimuli was found to arise within the midbrain posterior exterolateral nucleus (ELp). Whole-cell patch recordings from ELp neurons in vivo revealed three patterns of interpulse interval (IPI) tuning: low-pass neurons tuned to long intervals, high-pass neurons tuned to short intervals, and bandpass neurons tuned to intermediate intervals. Many neurons within each class also responded preferentially to either increasing or decreasing IPIs. Playback of electric signaling patterns recorded from freely behaving fish revealed that the IPI and direction tuning of ELp neurons resulted in selective responses to particular social communication displays characterized by distinct IPI patterns. The postsynaptic potential responses of many neurons indicated a combination of excitatory and inhibitory synaptic input, and the IPI tuning of ELp neurons was directly related to rate-dependent changes in the direction and amplitude of postsynaptic potentials. These results suggest that differences in the dynamics of short-term synaptic plasticity in excitatory and inhibitory pathways may tune central sensory neurons to particular temporal patterns of presynaptic activity. This may represent a general mechanism for the processing of behaviorally relevant stimulus information encoded into temporal patterns of activity by sensory neurons.

Loss of Anticodon Wobble Uridine Modifications Affects tRNALys Function and Protein Levels in Saccharomyces cerevisiae

PubMed Central

Klassen, Roland; Grunewald, Pia; Thüring, Kathrin L.; Eichler, Christian; Helm, Mark; Schaffrath, Raffael

2015-01-01

In eukaryotes, wobble uridines in the anticodons of tRNALys UUU, tRNAGlu UUC and tRNAGln UUG are modified to 5-methoxy-carbonyl-methyl-2-thio-uridine (mcm5s2U). While mutations in subunits of the Elongator complex (Elp1-Elp6), which disable mcm5 side chain formation, or removal of components of the thiolation pathway (Ncs2/Ncs6, Urm1, Uba4) are individually tolerated, the combination of both modification defects has been reported to have lethal effects on Saccharomyces cerevisiae. Contrary to such absolute requirement of mcm5s2U for viability, we demonstrate here that in the S. cerevisiae S288C-derived background, both pathways can be simultaneously inactivated, resulting in combined loss of tRNA anticodon modifications (mcm5U and s2U) without a lethal effect. However, an elp3 disruption strain displays synthetic sick interaction and synergistic temperature sensitivity when combined with either uba4 or urm1 mutations, suggesting major translational defects in the absence of mcm5s2U modifications. Consistent with this notion, we find cellular protein levels drastically decreased in an elp3uba4 double mutant and show that this effect as well as growth phenotypes can be partially rescued by excess of tRNALys UUU. These results may indicate a global translational or protein homeostasis defect in cells simultaneously lacking mcm5 and s2 wobble uridine modification that could account for growth impairment and mainly originates from tRNALys UUU hypomodification and malfunction. PMID:25747122

Loss of anticodon wobble uridine modifications affects tRNA(Lys) function and protein levels in Saccharomyces cerevisiae.

PubMed

Klassen, Roland; Grunewald, Pia; Thüring, Kathrin L; Eichler, Christian; Helm, Mark; Schaffrath, Raffael

2015-01-01

In eukaryotes, wobble uridines in the anticodons of tRNA(Lys)UUU, tRNA(Glu)UUC and tRNA(Gln)UUG are modified to 5-methoxy-carbonyl-methyl-2-thio-uridine (mcm5s2U). While mutations in subunits of the Elongator complex (Elp1-Elp6), which disable mcm5 side chain formation, or removal of components of the thiolation pathway (Ncs2/Ncs6, Urm1, Uba4) are individually tolerated, the combination of both modification defects has been reported to have lethal effects on Saccharomyces cerevisiae. Contrary to such absolute requirement of mcm5s2U for viability, we demonstrate here that in the S. cerevisiae S288C-derived background, both pathways can be simultaneously inactivated, resulting in combined loss of tRNA anticodon modifications (mcm5U and s2U) without a lethal effect. However, an elp3 disruption strain displays synthetic sick interaction and synergistic temperature sensitivity when combined with either uba4 or urm1 mutations, suggesting major translational defects in the absence of mcm5s2U modifications. Consistent with this notion, we find cellular protein levels drastically decreased in an elp3uba4 double mutant and show that this effect as well as growth phenotypes can be partially rescued by excess of tRNA(Lys)UUU. These results may indicate a global translational or protein homeostasis defect in cells simultaneously lacking mcm5 and s2 wobble uridine modification that could account for growth impairment and mainly originates from tRNA(Lys)UUU hypomodification and malfunction.

The effect of reminder letters on the uptake of an e-learning programme on dementia: a randomized trial in general practice.

PubMed

Waldorff, Frans Boch; Siersma, Volkert; Nielsen, Bente; Steenstrup, Annette Plesner; Bro, Flemming

2009-12-01

The aim of the present study was to evaluate whether three reminder letters mailed to GPs after dissemination of a Dementia Guideline increased the GPs' use of the corresponding e-learning programme (ELP). Single-blinded randomized trial among all GPs in Copenhagen Municipality from 1 November 2006 to 1 May 2007. A total of 15 of 320 GPs (4.7%) had a web-based logon during the study period. The intervention group had a significantly increased frequency of web-based logons (P = 0.0192) equivalent to a hazard ratio of 8.0 (95% CI: 1.03-66.1; P = 0.047). NNT was calculated to 22.2. We could not detect any significant differences in any of the secondary outcomes. Three reminder letters added to a nation-wide dissemination increased the probability for a GP logon in the ELP by a Factor 8. However, in total, only a small proportion used the ELP. Thus, further research is needed in order to consider future implementation strategies for Internet-based Continuous Medical Education activities among not primed GPs.

Double network physical gels from elastin-like polypeptide block copolymers: nanoscale control of thermoresponsive reinforcement

NASA Astrophysics Data System (ADS)

Glassman, Matthew; Olsen, Bradley

2014-03-01

Triblock copolymers with associative protein midblocks and thermoresponsive endblocks form shear thinning hydrogels with a low yield stress at low temperatures, but can be reinforced by a self-assembled network of the endblock aggregates. Here, we compare the use of bioengineered elastin-like polypeptides (ELPs) to synthetic poly(N-isopropylacrylamide) (PNIPAM) as endblocks to control the self-assembly of the reinforcing network. The temperature dependence of the mechanics of these hydrogels is a strong function of the domain size and morphology in the endblock network. Despite the architectural similarities, triblock ELP fusions and PNIPAM bioconjugates exhibit distinct reinforcement maxima at fixed block composition and polymer concentration, and these differences can be attributed to the nanostructural features of the two systems. Furthermore, in ELP fusions, the amino acid sequence can be readily modified to manipulate the solvation kinetics of the endblock domains. Finally, various endblocks have been combined to form triblock terpolymer hydrogels, demonstrating how the choice of thermoresponsive blocks can be used to tune the reinforcement of shear thinning hydrogels.

An ethical leadership program for nursing unit managers.

PubMed

Jeon, Sang Hee; Park, Mihyun; Choi, Kyungok; Kim, Mi Kyoung

2018-03-01

The aims of this study were to evaluate the effect of an ethical leadership program (ELP) on ethical leadership, organizational citizenship behavior (OCB), and job outcomes of nursing unit managers (UMs) and to examine changes in staff nurses' perception about UMs' EL, OCB, job outcomes, and ethical work environments (EWEs) post-ELP. A quasi-experimental (pre- and post-test design) study conducted six-month intervention (ELP) using self-reported UM survey (n=44), and staff nurses (n=158) were randomly extracted by two steps. The Korean version of Ethical Leadership at Work for UMs' self-ethical leadership, the Ethical Leadership Scale for staff nurses' perceived ethical leadership, a 19-item OCB scale, and six dimensions of the medium-sized Copenhagen Psychosocial Questionnaire II for job outcomes and EWEs were administered at baseline and post-intervention. UMs' ethical leadership scores differed significantly over time in people orientation (p=0.041) and concern for ethical leadership sustainability (p=0.002) adjusting for UM experience duration and nursing unit type. Total mean and level of power-sharing of ethical leadership among UMs with <5years of UM experience improved significantly over time. Of staff nurses' perception changes about UMs' ethical leadership, OCB, job outcomes, and EWEs, significant improvement over time appeared only in EWEs' work influence level (p=0.007). This study provides useful information for clinical ELP development and examining the program's effect on leadership skills and followers' outcomes. Program facilitation relies on practical training methods, participant motivation, and assessment outcome designs by controlling clinical confounding factors. Findings have implications as an attempt for intervention to promote competencies related to ethical leadership of nursing unit managers. Copyright © 2017 Elsevier Ltd. All rights reserved.

Development of a program for toric intraocular lens calculation considering posterior corneal astigmatism, incision-induced posterior corneal astigmatism, and effective lens position.

PubMed

Eom, Youngsub; Ryu, Dongok; Kim, Dae Wook; Yang, Seul Ki; Song, Jong Suk; Kim, Sug-Whan; Kim, Hyo Myung

2016-10-01

To evaluate the toric intraocular lens (IOL) calculation considering posterior corneal astigmatism, incision-induced posterior corneal astigmatism, and effective lens position (ELP). Two thousand samples of corneal parameters with keratometric astigmatism ≥ 1.0 D were obtained using bootstrap methods. The probability distributions for incision-induced keratometric and posterior corneal astigmatisms, as well as ELP were estimated from the literature review. The predicted residual astigmatism error using method D with an IOL add power calculator (IAPC) was compared with those derived using methods A, B, and C through Monte-Carlo simulation. Method A considered the keratometric astigmatism and incision-induced keratometric astigmatism, method B considered posterior corneal astigmatism in addition to the A method, method C considered incision-induced posterior corneal astigmatism in addition to the B method, and method D considered ELP in addition to the C method. To verify the IAPC used in this study, the predicted toric IOL cylinder power and its axis using the IAPC were compared with ray-tracing simulation results. The median magnitude of the predicted residual astigmatism error using method D (0.25 diopters [D]) was smaller than that derived using methods A (0.42 D), B (0.38 D), and C (0.28 D) respectively. Linear regression analysis indicated that the predicted toric IOL cylinder power and its axis had excellent goodness-of-fit between the IAPC and ray-tracing simulation. The IAPC is a simple but accurate method for predicting the toric IOL cylinder power and its axis considering posterior corneal astigmatism, incision-induced posterior corneal astigmatism, and ELP.

Grip strength is a predictor of bone mineral density among adolescent combat sport athletes.

PubMed

Nasri, Raouf; Hassen Zrour, Saoussen; Rebai, Haithem; Fadhel Najjar, Mohamed; Neffeti, Fadoua; Bergaoui, Naceur; Mejdoub, Hafedh; Tabka, Zouhair

2013-01-01

The aim of this study was firstly to investigate the correlation between bone parameters and grip strength (GS) in hands, explosive legs power (ELP), and hormonal parameters; second, to identify the most determinant variables of bone mineral density (BMD) among adolescent combat sport athletes. Fifty combat sport athletes aged 17.1 ± 0.2 year were compared with 30 sedentary subjects matched for age, height, and pubertal stage. For all subjects, the BMD in deferent sites associated with anthropometric parameters were measured by dual-energy X-ray absorptiometry. The growth hormone (GH) and testosterone (TESTO) concentrations were tested. The GS in dominant (GSDA) and nondominant arms (GSNDA) and ELP were evaluated. All BMD measured were greater in athletes than in sedentary group (p<0.01). The GS and ELP showed higher values in athletes than in sedentary group (p<0.01). The BMD in all sites were correlated with weight, but without correlation with height. The GSNDA and ELP were significantly correlated with BMD of both spine and legs. The GH was correlated with the BMD of whole body and spine (p<0.05). The TESTO was only correlated with BMD of the arms (p<0.01). The best predictor of BMD measurements is GSNDA. This study has proved the osteogenic effect of combat sports practice, especially judo and karate kyokushinkai. Therefore, children and adolescent should be encouraged to participate in combat sport. Moreover, it suggested that the best model predicting BMD in different sites among adolescent combat sports athletes was the GSNDA. Copyright © 2013 The International Society for Clinical Densitometry. Published by Elsevier Inc. All rights reserved.

Error induced by the estimation of the corneal power and the effective lens position with a rotationally asymmetric refractive multifocal intraocular lens

PubMed Central

Piñero, David P.; Camps, Vicente J.; Ramón, María L.; Mateo, Verónica; Pérez-Cambrodí, Rafael J.

2015-01-01

AIM To evaluate the prediction error in intraocular lens (IOL) power calculation for a rotationally asymmetric refractive multifocal IOL and the impact on this error of the optimization of the keratometric estimation of the corneal power and the prediction of the effective lens position (ELP). METHODS Retrospective study including a total of 25 eyes of 13 patients (age, 50 to 83y) with previous cataract surgery with implantation of the Lentis Mplus LS-312 IOL (Oculentis GmbH, Germany). In all cases, an adjusted IOL power (PIOLadj) was calculated based on Gaussian optics using a variable keratometric index value (nkadj) for the estimation of the corneal power (Pkadj) and on a new value for ELP (ELPadj) obtained by multiple regression analysis. This PIOLadj was compared with the IOL power implanted (PIOLReal) and the value proposed by three conventional formulas (Haigis, Hoffer Q and Holladay I). RESULTS PIOLReal was not significantly different than PIOLadj and Holladay IOL power (P>0.05). In the Bland and Altman analysis, PIOLadj showed lower mean difference (-0.07 D) and limits of agreement (of 1.47 and -1.61 D) when compared to PIOLReal than the IOL power value obtained with the Holladay formula. Furthermore, ELPadj was significantly lower than ELP calculated with other conventional formulas (P<0.01) and was found to be dependent on axial length, anterior chamber depth and Pkadj. CONCLUSION Refractive outcomes after cataract surgery with implantation of the multifocal IOL Lentis Mplus LS-312 can be optimized by minimizing the keratometric error and by estimating ELP using a mathematical expression dependent on anatomical factors. PMID:26085998

Error induced by the estimation of the corneal power and the effective lens position with a rotationally asymmetric refractive multifocal intraocular lens.

PubMed

Piñero, David P; Camps, Vicente J; Ramón, María L; Mateo, Verónica; Pérez-Cambrodí, Rafael J

2015-01-01

To evaluate the prediction error in intraocular lens (IOL) power calculation for a rotationally asymmetric refractive multifocal IOL and the impact on this error of the optimization of the keratometric estimation of the corneal power and the prediction of the effective lens position (ELP). Retrospective study including a total of 25 eyes of 13 patients (age, 50 to 83y) with previous cataract surgery with implantation of the Lentis Mplus LS-312 IOL (Oculentis GmbH, Germany). In all cases, an adjusted IOL power (PIOLadj) was calculated based on Gaussian optics using a variable keratometric index value (nkadj) for the estimation of the corneal power (Pkadj) and on a new value for ELP (ELPadj) obtained by multiple regression analysis. This PIOLadj was compared with the IOL power implanted (PIOLReal) and the value proposed by three conventional formulas (Haigis, Hoffer Q and Holladay I). PIOLReal was not significantly different than PIOLadj and Holladay IOL power (P>0.05). In the Bland and Altman analysis, PIOLadj showed lower mean difference (-0.07 D) and limits of agreement (of 1.47 and -1.61 D) when compared to PIOLReal than the IOL power value obtained with the Holladay formula. Furthermore, ELPadj was significantly lower than ELP calculated with other conventional formulas (P<0.01) and was found to be dependent on axial length, anterior chamber depth and Pkadj. Refractive outcomes after cataract surgery with implantation of the multifocal IOL Lentis Mplus LS-312 can be optimized by minimizing the keratometric error and by estimating ELP using a mathematical expression dependent on anatomical factors.

Multicolor Luminescence from Conjugates of Genetically Encoded Elastin-like Polymers and Terpyridine-Lanthanides

DOE PAGES

Ghosh, Koushik; Balog, Eva Rose M.; Kahn, Jennifer L.; ...

2015-08-20

Functional hybrid materials with optically active metal-ligand moieties embedded within a polymer matrix have a great potential in (bio)materials science, including applications in light-emitting diode devices. Here, we report a simple strategy to incorporate terpyridine derivatives into the side chains of elastin-like polymers (ELPs). The further binding of trivalent lanthanide ions with the terpyridine ligands generated an array of photoluminescence ranging from the visible to the near-infrared regions. Lastly, as thin films, these ELP-based optical materials also exhibited distinct morphologies that depend upon the temperature of the aqueous solutions from which the hybrid polymers were spin coated or drop cast.

Engineering protein scaffolds for protein separation, biocatalysis and nanotechnology applications

NASA Astrophysics Data System (ADS)

Liu, Fang

Globally, there is growing appreciation for developing a sustainable economy that uses eco-efficient bio-processes. Biotechnology provides an increasing range of tools for industry to help reduce cost and improve environmental performance. Inspired by the naturally evolved machineries of protein scaffolds and their binding ligands, synthetic protein scaffolds were engineered based on cohesin-dockerin interactions and metal chelating peptides to tackle the challenges and make improvements in three specific areas: (1) protein purification, (2) biofuel cells, and (3) nanomaterial synthesis. The first objective was to develop efficient and cost-effective non-chromatographic purification processes to purify recombinant proteins in an effort to meet the dramatically growing market of protein drugs. In our design, the target protein was genetically fused with a dockerin domain from Clostridium thermocellum and direct purification and recovery was achieved using thermo-responsive elastin-like polypeptide (ELP) scaffold containing the cohesin domain from the same species. By exploiting the highly specific interaction between the dockerin and cohesin domain and the reversible aggregation property of ELP, highly purified and active dockerin-tagged proteins, such as endoglucanase CelA, chloramphenicol acetyl transferase (CAT) and enhanced green fluorescence protein (EGFP), were recovered directly from crude cell extracts in a single purification step with yields achieving over 90%. Incorporation of a self-cleaving intein domain enabled rapid removal of the affinity tag from the target proteins by another cycle of thermal precipitation. The purification cost can be further reduced by regenerating and recycling the ELP-cohesin capturing scaffolds. However, due to the high binding affinity between cohesin and dockerin domains, the bound dockerin-intein tag cannot be completely disassociated from ELP-cohesin scaffold after binding. Therefore, a truncated dockerin with the calcium-coordinating function impaired was used in replace of the original full length dockerin domain. The truncated dockerin domain maintained its functionality as an effective affinity tag, and efficient EDTA mediated dissociation of the bound dockerin-intein tag was also realized. The regenerated ELP capturing scaffold was reused for additional purification cycles without any decrease in efficiency. The second objective was to assemble biocatalysts for biofuel cells. Three beta-NAD dependent dehydrogenases, alcohol dehydrogenase (ADH), formaldehyde dehydrogenase (FALDH) and formate dehydrogenase (FDH), were site-specifically co-localized onto the scaffolds displayed on the yeast surface based on the high-affinity interactions between three orthogonal cohesin/dockerin pairs. The assembled multi-enzyme cascades, which can completely convert methanol to CO2, showed improved production yield compared with that of the non-complexed enzyme mixture, indicating efficient substrate channeling among the three enzymes. This strategy can be easily extended to other complex cascade reactions for enzymatic fuel cell applications. To further explore the role of biotechnology toward environmental sustainability, Escherichia coli was engineered to express phytochelatin synthase, which converted glutathione into the metal-binding peptide phytochelatin (PC). PCs served as peptide scaffolds and mediated synthesis of CdS nanocrystals. This approach may be generalized to guide the in vitro self-assembly of a wide range of nanocrystals with different compositions and sizes.

Evolutionary learning processes as the foundation for behaviour change.

PubMed

Crutzen, Rik; Peters, Gjalt-Jorn Ygram

2018-03-01

We argue that the active ingredients of behaviour change interventions, often called behaviour change methods (BCMs) or techniques (BCTs), can usefully be placed on a dimension of psychological aggregation. We introduce evolutionary learning processes (ELPs) as fundamental building blocks that are on a lower level of psychological aggregation than BCMs/BCTs. A better understanding of ELPs is useful to select the appropriate BCMs/BCTs to target determinants of behaviour, or vice versa, to identify potential determinants targeted by a given BCM/BCT, and to optimally translate them into practical applications. Using these insights during intervention development may increase the likelihood of developing effective interventions - both in terms of behaviour change as well as maintenance of behaviour change.

The Effects of NBF Gingival Gel Application in the Treatment of the Erosive Lichen Planus: Case Report.

PubMed

Popovska, Mirjana; Fidovski, Jasmin; Mindova, Sonja; Dirjanska, Katerina; Ristoska, Stevica; Stefanovska, Emilija; Radojkova-Nikolovska, Vera; Mitic, Kristina; Rusevska, Biljana

2016-03-15

The therapy of erosive lichen planus (ELP) has been particular problem in the treatment of oral lesions. This case of ELP in male patient 29 years old was treated with topic application of the NBF gingival gel, three times a day after meal, previously rinsed with Clorhexidine gluconate 0.12%. After 5 days of treatment, initial improvements were recorded, and after two weeks of application of the NBF gingival gel we observed significant improvement. Clinical monitoring after the fifth day showed mild epithelialization of the eroded mucosa, yet still present erythematous base of the lesion. After the second week the erythema area was significantly reduced and the eroded surfaces of the mucosa were minimal, measured less than 0.5 mm. After the third week there were no erosions to detect on the oral mucosa, yet still present vague redness, which completely pulled after the fourth week. Treatment ended after the fifth week when the topical application of the NBF gingival gel was terminated, and therapy was done, and clinically achieved effects remained stable even after the third month of the treatment. Topic application of the NBF gingival gel with ELP patients showed positive clinical effects in relatively short time period.

The Effects of NBF Gingival Gel Application in the Treatment of the Erosive Lichen Planus: Case Report

PubMed Central

Popovska, Mirjana; Fidovski, Jasmin; Mindova, Sonja; Dirjanska, Katerina; Ristoska, Stevica; Stefanovska, Emilija; Radojkova-Nikolovska, Vera; Mitic, Kristina; Rusevska, Biljana

2016-01-01

The therapy of erosive lichen planus (ELP) has been particular problem in the treatment of oral lesions. This case of ELP in male patient 29 years old was treated with topic application of the NBF gingival gel, three times a day after meal, previously rinsed with Clorhexidine gluconate 0.12%. After 5 days of treatment, initial improvements were recorded, and after two weeks of application of the NBF gingival gel we observed significant improvement. Clinical monitoring after the fifth day showed mild epithelialization of the eroded mucosa, yet still present erythematous base of the lesion. After the second week the erythema area was significantly reduced and the eroded surfaces of the mucosa were minimal, measured less than 0.5 mm. After the third week there were no erosions to detect on the oral mucosa, yet still present vague redness, which completely pulled after the fourth week. Treatment ended after the fifth week when the topical application of the NBF gingival gel was terminated, and therapy was done, and clinically achieved effects remained stable even after the third month of the treatment. Topic application of the NBF gingival gel with ELP patients showed positive clinical effects in relatively short time period. PMID:27275352

Bright lights, big city: influences of ecological light pollution on reciprocal stream-riparian invertebrate fluxes.

PubMed

Meyer, Lars A; Sullivan, S Mazeika P

2013-09-01

Cities produce considerable ecological light pollution (ELP), yet the effects of artificial night lighting on biological communities and ecosystem function have not been fully explored. From June 2010 to June 2011, we surveyed aquatic emergent insects, riparian arthropods entering the water, and riparian spiders of the family Tetragnathidae at nine stream reaches representing common ambient ELP levels of Columbus, Ohio, USA, streams (low, 0.1-0.5 lux; moderate, 0.6-2.0 lux; high, 2.1-4.0 lux). In August 2011, we experimentally increased light levels at the low- and moderate-treatment reaches to 10-12 lux to represent urban streams exposed to extremely high levels of ELP. Although season exerted the dominant influence on invertebrate fluxes over the course of the year, when analyzed by season, we found that light strongly influenced multiple invertebrate responses. The experimental light addition resulted in a 44% decrease in tetragnathid spider density (P = 0.035), decreases of 16% in family richness (P = 0.040) and 76% in mean body size (P = 0.022) of aquatic emergent insects, and a 309% increase in mean body size of terrestrial arthropods (P = 0.015). Our results provide evidence that artificial light sources can alter community structure and ecosystem function in streams via changes in reciprocal aquatic-terrestrial fluxes of invertebrates.

Tunable elastin-like polypeptide hollow sphere as a high payload and controlled delivery gene depot.

PubMed

Dash, Biraja C; Mahor, Sunil; Carroll, Oliver; Mathew, Asha; Wang, Wenxin; Woodhouse, Kimberly A; Pandit, Abhay

2011-06-30

Self-assembly driven processes can be utilized to produce a variety of nanostructures useful for various in vitro and in vivo applications. Characteristics such as size, stability, biocompatibility, high therapeutic loading and controlled delivery of these nanostructures are particularly crucial in relation to in vivo applications. In this study, we report the fabrication of tunable monodispersed elastin-like polypeptide (ELP) hollow spheres of 100, 300, 500 and 1000 nm by exploiting the self-assembly property and net positive charge of ELP. The microbial transglutaminase (mTGase) cross-linking provided robustness and stability to the hollow spheres while maintaining surface functional groups for further modifications. The resulting hollow spheres showed a higher loading efficiency of plasmid DNA (pDNA) by using polyplex (~70 μg pDNA/mg of hollow sphere) than that of self-assembled ELP particles and demonstrated controlled release triggered by protease and elastase. Moreover, polyplex-loaded hollow spheres showed better cell viability than polyplex alone and yielded higher luciferase expression by providing protection against endosomal degradation. Overall, the monodispersed, tunable hollow spheres with a capability of post-functionalization can provide an exciting new opportunity for use in a range of therapeutic and diagnostic applications. Copyright © 2011 Elsevier B.V. All rights reserved.

Systemic therapy for vulval Erosive Lichen Planus (the 'hELP' trial): study protocol for a randomised controlled trial.

PubMed

Simpson, Rosalind C; Murphy, Ruth; Bratton, Daniel J; Sydes, Matthew R; Wilkes, Sally; Nankervis, Helen; Dowey, Shelley; Thomas, Kim S

2016-01-04

Erosive lichen planus affecting the vulva (ELPV) is a relatively rare, chronic condition causing painful raw areas in the vulvovaginal region. Symptoms are pain and burning, which impact upon daily living. There is paucity of evidence regarding therapy. A 2012 Cochrane systematic review found no randomised controlled trials (RCTs) in this field. Topically administered corticosteroids are the accepted first-line therapy: however, there is uncertainty as to which second-line treatments to use. Several systemic agents have been clinically noted to show promise for ELPV refractory to topically administered corticosteroids but there is no RCT evidence to support these. The 'hELP' study is a RCT with an internal pilot phase designed to provide high-quality evidence. The objective is to test whether systemic therapy in addition to standard topical therapy is a beneficial second-line treatment for ELPV. Adjunctive systemic therapies used are hydroxychloroquine, methotrexate, mycophenolate mofetil and prednisolone. Topical therapy plus a short course of prednisolone given orally is considered the comparator intervention. The trial is a four-armed, open-label, pragmatic RCT which uses a blinded independent clinical assessor. To provide 80 % power for each comparison, 96 participants are required in total. The pilot phase aims to recruit 40 participants. The primary clinical outcome is the proportion of patients achieving treatment success at 6 months. 'Success' is defined by a composite measure of Patient Global Assessment score of 0 or 1 on a 4-point scale plus improvement from baseline on clinical photographs scored by a clinician blinded to treatment allocation. Secondary clinical outcomes include 6-month assessment of: (1) Reduction in pain/soreness; (2) Global assessment of disease; (3) Response at other affected mucosal sites; (4) Hospital Anxiety and Depression Scale scores; (5) Sexual function; (6) Health-related quality of life using 'Short Form 36' and 'Skindex-29' questionnaires; (7) Days of topical steroid use; (8) Treatment satisfaction; (9) Discontinuation of medications due to treatment failure; (10) Per participant cost of intervention in each treatment group. Adverse events will also be reported. 'hELP' is the first RCT to address second-line treatment of ELPV. The trial has encountered unique methodological challenges and has required collaborative efforts of the UK Dermatology Clinical Trials Network alongside expert clinicians. ISRCTN 81883379 . Date of registration 12 June 2014.

40 CFR 52.2309 - Emissions inventories.

Code of Federal Regulations, 2013 CFR

2013-07-01

.../Galveston (HGA), Beaumont/Port Arthur (BPA), El Paso (ELP), and Dallas/Fort Worth (DFW) ozone nonattainment... Counties; the BPA nonattainment area is classified as Serious and includes Hardin, Jefferson, and Orange...

40 CFR 52.2309 - Emissions inventories.

Code of Federal Regulations, 2011 CFR

2011-07-01

.../Galveston (HGA), Beaumont/Port Arthur (BPA), El Paso (ELP), and Dallas/Fort Worth (DFW) ozone nonattainment... Counties; the BPA nonattainment area is classified as Serious and includes Hardin, Jefferson, and Orange...

40 CFR 52.2309 - Emissions inventories.

Code of Federal Regulations, 2010 CFR

2010-07-01

.../Galveston (HGA), Beaumont/Port Arthur (BPA), El Paso (ELP), and Dallas/Fort Worth (DFW) ozone nonattainment... Counties; the BPA nonattainment area is classified as Serious and includes Hardin, Jefferson, and Orange...

40 CFR 52.2309 - Emissions inventories.

Code of Federal Regulations, 2012 CFR

2012-07-01

.../Galveston (HGA), Beaumont/Port Arthur (BPA), El Paso (ELP), and Dallas/Fort Worth (DFW) ozone nonattainment... Counties; the BPA nonattainment area is classified as Serious and includes Hardin, Jefferson, and Orange...

40 CFR 52.2309 - Emissions inventories.

Code of Federal Regulations, 2014 CFR

2014-07-01

.../Galveston (HGA), Beaumont/Port Arthur (BPA), El Paso (ELP), and Dallas/Fort Worth (DFW) ozone nonattainment... Counties; the BPA nonattainment area is classified as Serious and includes Hardin, Jefferson, and Orange...

Extensive excitatory network interactions shape temporal processing of communication signals in a model sensory system.

PubMed

Ma, Xiaofeng; Kohashi, Tsunehiko; Carlson, Bruce A

2013-07-01

Many sensory brain regions are characterized by extensive local network interactions. However, we know relatively little about the contribution of this microcircuitry to sensory coding. Detailed analyses of neuronal microcircuitry are usually performed in vitro, whereas sensory processing is typically studied by recording from individual neurons in vivo. The electrosensory pathway of mormyrid fish provides a unique opportunity to link in vitro studies of synaptic physiology with in vivo studies of sensory processing. These fish communicate by actively varying the intervals between pulses of electricity. Within the midbrain posterior exterolateral nucleus (ELp), the temporal filtering of afferent spike trains establishes interval tuning by single neurons. We characterized pairwise neuronal connectivity among ELp neurons with dual whole cell recording in an in vitro whole brain preparation. We found a densely connected network in which single neurons influenced the responses of other neurons throughout the network. Similarly tuned neurons were more likely to share an excitatory synaptic connection than differently tuned neurons, and synaptic connections between similarly tuned neurons were stronger than connections between differently tuned neurons. We propose a general model for excitatory network interactions in which strong excitatory connections both reinforce and adjust tuning and weak excitatory connections make smaller modifications to tuning. The diversity of interval tuning observed among this population of neurons can be explained, in part, by each individual neuron receiving a different complement of local excitatory inputs.

YAP-dependent Mechanotransduction is Required for Proliferation and Migration on Native-like Substrate Topography

PubMed Central

Mascharak, Shamik; Benitez, Patrick L.; Proctor, Amy C.; Madl, Christopher M.; Hu, Kenneth H.; Dewi, Ruby E.; Butte, Manish J.; Heilshorn, Sarah C.

2017-01-01

Native vascular extracellular matrices (vECM) consist of elastic fibers that impart varied topographical properties, yet most in vitro models designed to study the effects of topography on cell behavior are not representative of native architecture. Here, we engineer an electrospun elastin-like protein (ELP) system with independently tunable, vECM-mimetic topography and demonstrate that increasing topographical variation causes loss of endothelial cell-cell junction organization. This loss of VE-cadherin signaling and increased cytoskeletal contractility on more topographically varied ELP substrates in turn promote YAP activation and nuclear translocation, resulting in significantly increased endothelial cell migration and proliferation. Our findings identify YAP as a required signaling factor through which fibrous substrate topography influences cell behavior and highlights topography as a key design parameter for engineered biomaterials. PMID:27889666

High performance fuel electrodes fabricated by electroless plating of copper on BaZr0.8Ce0.1Y0.1O3-δ proton-conducting ceramic

NASA Astrophysics Data System (ADS)

Patki, Neil S.; Way, J. Douglas; Ricote, Sandrine

2017-10-01

The stability of copper at high temperatures in reducing and hydrocarbon-containing atmospheres makes it a good candidate for fabricating fuel electrodes on proton-conducting ceramics, such as BaZr0.9-xCexY0.1O3-δ (BZCY). In this work, the electrochemical performance of Cu-based electrodes fabricated by electroless plating (ELP) on BaZr0.8Ce0.1Y0.1O3-δ is studied with impedance spectroscopy. Three activation catalysts (Pd, Ru, and Cu) are investigated and ELP is compared to a commercial Cu paste (ESL 2312-G) for electrode fabrication. The area specific resistances (ASR) for Pd, Ru, and Cu activations at 700 °C in moist 5% H2 in Ar are 2.1, 3.2, and 13.4 Ω cm2, respectively. That is a 1-2 orders of magnitude improvement over the commercial Cu paste (192 Ω cm2). Furthermore, the ASR has contributions from electrode processes and charge transfer at the electrode/electrolyte interface. Additionally, the morphology of the as-fabricated electrode is unaffected by the activation catalyst. However, heat treatment at 750 °C in H2 for 24 h leads to sintering and large reorganization of the electrode fabricated with Cu activation (micron sized pores seen in the tested sample), while Pd and Ru activations are immune to such reorganization. Thus, Pd and Ru are identified as candidates for future work with improvements to charge transfer required for the former, and better electrode processes required for the latter.

YAP-dependent mechanotransduction is required for proliferation and migration on native-like substrate topography.

PubMed

Mascharak, Shamik; Benitez, Patrick L; Proctor, Amy C; Madl, Christopher M; Hu, Kenneth H; Dewi, Ruby E; Butte, Manish J; Heilshorn, Sarah C

2017-01-01

Native vascular extracellular matrices (vECM) consist of elastic fibers that impart varied topographical properties, yet most in vitro models designed to study the effects of topography on cell behavior are not representative of native architecture. Here, we engineer an electrospun elastin-like protein (ELP) system with independently tunable, vECM-mimetic topography and demonstrate that increasing topographical variation causes loss of endothelial cell-cell junction organization. This loss of VE-cadherin signaling and increased cytoskeletal contractility on more topographically varied ELP substrates in turn promote YAP activation and nuclear translocation, resulting in significantly increased endothelial cell migration and proliferation. Our findings identify YAP as a required signaling factor through which fibrous substrate topography influences cell behavior and highlights topography as a key design parameter for engineered biomaterials. Copyright © 2016 Elsevier Ltd. All rights reserved.

A familial pericentric inversion of chromosome 11 associated with a microdeletion of 163 kb and microduplication of 288 kb at 11p13 and 11q22.3 without aniridia or eye anomalies.

PubMed

Balay, Lara; Totten, Ellen; Okada, Luna; Zell, Sidney; Ticho, Benjamin; Israel, Jeannette; Kogan, Jillene

2016-01-01

Interstitial deletions of 11p13 involving MPPED2, DCDC5, DCDC1, DNAJC24, IMMP1L, and ELP4 are previously reported to have downstream transcriptional effects on the expression of PAX6, due to a downstream regulatory region (DRR). Currently, no clear genotype-phenotype correlations have been established allowing for conclusive information regarding the exact location of the PAX6 DRR, though its location has been approximated in mouse models to be within the Elp4 gene. Of the clinical reports currently published examining patients with intact PAX6 genes but harboring deletions identified in genes downstream of PAX6, 100% indicate phenotypes which include aniridia, whereas approximately half report additional eye deformities, autism, or intellectual disability. In this clinical report, we present a 12-year-old male patient, his brother, and mother with pericentric inversions of chromosome 11 associated with submicroscopic interstitial deletions of 11p13 and duplications of 11q22.3. The inversions were identified by standard cytogenetic analysis; microarray and FISH detected the chromosomal imbalance. The patient's phenotype includes intellectual disability, speech abnormalities, and autistic behaviors, but interestingly neither the patient, his brother, nor mother have aniridia or other eye anomalies. To the best of our knowledge, these findings in three family members represent the only reported cases with 11p13 deletions downstream of PAX6 not demonstrating phenotypic characteristics of aniridia or abnormal eye development. Although none of the deleted genes are obvious candidates for the patient's phenotype, the absence of aniridia in the presence of this deletion in all three family members further delineates the location of the DRR for PAX6. © 2015 Wiley Periodicals, Inc.

Sequence-Independent Cloning and Post-Translational Modification of Repetitive Protein Polymers through Sortase and Sfp-Mediated Enzymatic Ligation.

PubMed

Ott, Wolfgang; Nicolaus, Thomas; Gaub, Hermann E; Nash, Michael A

2016-04-11

Repetitive protein-based polymers are important for many applications in biotechnology and biomaterials development. Here we describe the sequential additive ligation of highly repetitive DNA sequences, their assembly into genes encoding protein-polymers with precisely tunable lengths and compositions, and their end-specific post-translational modification with organic dyes and fluorescent protein domains. Our new Golden Gate-based cloning approach relies on incorporation of only type IIS BsaI restriction enzyme recognition sites using PCR, which allowed us to install ybbR-peptide tags, Sortase c-tags, and cysteine residues onto either end of the repetitive gene polymers without leaving residual cloning scars. The assembled genes were expressed in Escherichia coli and purified using inverse transition cycling (ITC). Characterization by cloud point spectrophotometry, and denaturing polyacrylamide gel electrophoresis with fluorescence detection confirmed successful phosphopantetheinyl transferase (Sfp)-mediated post-translational N-terminal labeling of the protein-polymers with a coenzyme A-647 dye (CoA-647) and simultaneous sortase-mediated C-terminal labeling with a GFP domain containing an N-terminal GG-motif in a one-pot reaction. In a further demonstration, we installed an N-terminal cysteine residue into an elastin-like polypeptide (ELP) that was subsequently conjugated to a single chain poly(ethylene glycol)-maleimide (PEG-maleimide) synthetic polymer, noticeably shifting the ELP cloud point. The ability to straightforwardly assemble repetitive DNA sequences encoding ELPs of precisely tunable length and to post-translationally modify them specifically at the N- and C- termini provides a versatile platform for the design and production of multifunctional smart protein-polymeric materials.

Light Scattering Study of Mixed Micelles Made from Elastin-Like Polypeptide Linear Chains and Trimers

NASA Astrophysics Data System (ADS)

Terrano, Daniel; Tsuper, Ilona; Maraschky, Adam; Holland, Nolan; Streletzky, Kiril

Temperature sensitive nanoparticles were generated from a construct (H20F) of three chains of elastin-like polypeptides (ELP) linked to a negatively charged foldon domain. This ELP system was mixed at different ratios with linear chains of ELP (H40L) which lacks the foldon domain. The mixed system is soluble at room temperature and at a transition temperature (Tt) will form swollen micelles with the hydrophobic linear chains hidden inside. This system was studied using depolarized dynamic light scattering (DDLS) and static light scattering (SLS) to determine the size, shape, and internal structure of the mixed micelles. The mixed micelle in equal parts of H20F and H40L show a constant apparent hydrodynamic radius of 40-45 nm at the concentration window from 25:25 to 60:60 uM (1:1 ratio). At a fixed 50 uM concentration of the H20F, varying H40L concentration from 5 to 80 uM resulted in a linear growth in the hydrodynamic radius from about 11 to about 62 nm, along with a 1000-fold increase in VH signal. A possible simple model explaining the growth of the swollen micelles is considered. Lastly, the VH signal can indicate elongation in the geometry of the particle or could possibly be a result from anisotropic properties from the core of the micelle. SLS was used to study the molecular weight, and the radius of gyration of the micelle to help identify the structure and morphology of mixed micelles and the tangible cause of the VH signal.

Influence on photosynthesis of starlight, moonlight, planetlight, and light pollution (reflections on photosynthetically active radiation in the universe).

PubMed

Raven, J A; Cockell, C S

2006-08-01

Photosynthesis on Earth can occur in a diversity of organisms in the photosynthetically active radiation (PAR) range of 10 nmol of photons m(-2) s(-1) to 8 mmol of photons m(-2) s(-1). Similar considerations would probably apply to photosynthetic organisms on Earth-like planets (ELPs) in the continuously habitable zone of other stars. On Earth, starlight PAR is inadequate for photosynthetically supported growth. An increase in starlight even to reach the minimum theoretical levels to allow for photosynthesis would require a universe that was approximately ten million times older, or with a ten million times greater density of stars, than is the case for the present universe. Photosynthesis on an ELP using PAR reflected from a natural satellite with the same size as our Moon, but at the Roche limit, could support a low rate of photosynthesis at full Moon. Photosynthesis on an ELP-like satellite of a Jupiter-sized planet using light reflected from the planet could be almost 1% of the rate in full sunlight on Earth when the planet was full. These potential contributions to photosynthesis require that the contribution is compared with the rate of photosynthesis driven by direct radiation from the star. Light pollution on Earth only energizes photosynthesis by organisms that are very close to the light source. However, effects of light pollution on photosynthesis can be more widespread if the photosynthetic canopy is retained for more of the year, caused by effects on photoperiodism, with implications for the influence of civilizations on photosynthesis.

Trimethylamine N-oxide stabilizes proteins via a distinct mechanism compared with betaine and glycine.

PubMed

Liao, Yi-Ting; Manson, Anthony C; DeLyser, Michael R; Noid, William G; Cremer, Paul S

2017-03-07

We report experimental and computational studies investigating the effects of three osmolytes, trimethylamine N -oxide (TMAO), betaine, and glycine, on the hydrophobic collapse of an elastin-like polypeptide (ELP). All three osmolytes stabilize collapsed conformations of the ELP and reduce the lower critical solution temperature (LSCT) linearly with osmolyte concentration. As expected from conventional preferential solvation arguments, betaine and glycine both increase the surface tension at the air-water interface. TMAO, however, reduces the surface tension. Atomically detailed molecular dynamics (MD) simulations suggest that TMAO also slightly accumulates at the polymer-water interface, whereas glycine and betaine are strongly depleted. To investigate alternative mechanisms for osmolyte effects, we performed FTIR experiments that characterized the impact of each cosolvent on the bulk water structure. These experiments showed that TMAO red-shifts the OH stretch of the IR spectrum via a mechanism that was very sensitive to the protonation state of the NO moiety. Glycine also caused a red shift in the OH stretch region, whereas betaine minimally impacted this region. Thus, the effects of osmolytes on the OH spectrum appear uncorrelated with their effects upon hydrophobic collapse. Similarly, MD simulations suggested that TMAO disrupts the water structure to the least extent, whereas glycine exerts the greatest influence on the water structure. These results suggest that TMAO stabilizes collapsed conformations via a mechanism that is distinct from glycine and betaine. In particular, we propose that TMAO stabilizes proteins by acting as a surfactant for the heterogeneous surfaces of folded proteins.

Collagen like peptide bioconjugates for targeted drug delivery applications

NASA Astrophysics Data System (ADS)

Luo, Tianzhi

Collagen is the most abundant protein in mammals, and there has been long-standing interest in understanding and controlling collagen assembly in the design of new materials. Collagen-like peptides (CLP), also known as collagen-mimetic peptides (CMP), are short synthetic peptides which mimic the triple helical conformation of native collagens. In the past few decades, collagen like peptides and their conjugated hybrids have become a new class of biomaterials that possesses unique structures and properties. In addition to traditional applications of using CLPs to decipher the role of different amino acid residues and tripeptide motifs in stabilizing the collagen triple helix and mimicking collagen fibril formation, with the introduction of specific interactions including electrostatic interactions, pi-pi stacking interaction and metal-ligand coordination, a variety of artificial collagen-like peptides with well-defined sequences have been designed to create higher order assemblies with specific biological functions. The CLPs have also been widely used as bioactive domains or physical cross-linkers to fabricate hydrogels, which have shown potential to improve cell adhesion, proliferation and ECM macromolecule production. Despite this widespread use, the utilization of CLPs as domains in stimuli responsive bioconjugates represents a relatively new area for the development of functional polymeric materials. In this work, a new class of thermoresponsive diblock conjugates, containing collagen-like peptides and a thermoresponsive polymer, namely poly(diethylene glycol methyl ether methacrylate) (PDEGMEMA), is introduced. The CLP domain maintains its triple helix conformation after conjugation with the polymer. The engineered LCST of these conjugates has enabled temperature-induced assembly under aqueous conditions, at physiologically relevant temperatures, into well-defined vesicles with diameters of approximately 50-200 nm. The formation of nanostructures was driven by the coil/globule conformational transition of the PDEGMEMA building block above its LCST with stabilization of the nanostructures by the hydrophilic CLP. To the best of our knowledge, this is the first report on such assembled nanostructures from collagen-like peptide containing copolymers. Due to the strong propensity for CLPs to bind to natural collagen via strand invasion processes, these nanosized vesicles may be used as drug carriers for targeted delivery. In addition to synthetic polymers, the collagen like peptide is then conjugated with a thermoresponsive elastin-like peptide (ELP). The resulting ELP-CLP diblock conjugates show a remarkable reduction in the inverse transition temperature of the ELP domain, attributed to the anchoring effect of the CLP triple helix. The lower transition temperature of the conjugate enables facile formation of well-defined vesicles at physiological temperature and the unexpected resolubilization of the vesicles at elevated temperatures upon unfolding of the CLP domain. Given the ability of CLPs to modify collagens, this work provides not only a simple and versatile avenue for controlling the inverse transition behavior of elastin-like peptides, but also suggest future opportunities for these thermoresponsive nanostructures in biologically relevant environments. In the last section, the potential of using the ELP-CLP nanoparticles as drug delivery vehicles for targeting collagen containing matrices is evaluated. A sustained release of clinically relevant amount of encapsulated modelled drug is achieved within three weeks, followed by a thermally controlled burst release. As expected, the ELP-CLP nanoparticles show strong retention on collagen substrate, via specific binding through collagen triple helix hybridization. Additionally, cell viability and proliferation studies using fibroblasts and chondrocytes suggest the nanoparticles are non-cytotoxic. Additionally, almost no TNF-alpha expression from macrophages is observed, suggesting that the nanoparticles do not initiate inflammatory response. Endowed with specific collagen binding, controlled thermoresponsiveness, excellent cytocompatibility, and non-immune responsiveness, we believe the ELP-CLP nanoparticles are promising candidates as drug delivery vehicles for targeting collagen containing matrices. Considering the critical role of collagens in extracellular matrix and the unique ability of the CLP to target native collagens, our work offers significant opportunities for the design of collagen-like peptides and their bioconjugates for targeted application in the biomedical arena.

A novel method for simultaneous purification and immobilization of a xylanase-lichenase chimera via SpyTag/SpyCatcher spontaneous reaction.

PubMed

Lin, Yuanqing; Jin, Wenhui; Wang, Jindan; Cai, Zhengwen; Wu, Shuyu; Zhang, Guangya

2018-08-01

We generated a bifunctional enzyme chimera containing the xylanase and lichenase coupled with SpyTag between them. Meanwhile, we generated another chimera containing SpyCatcher and elastin-like polypeptides (ELPs). As ELPs could bond to the xylanase-lichenase chimera through SpyTag/SpyCatcher spontaneous reaction in mild condition, which would lead to the formation of a 3-arm star multifunctional chimera. We purified the xylanase-lichenase by the non-chromatographic purification tag of ELPs. Interestingly, 57.5% of the xylanase and 47.2% of the lichenase in chimera self-assembled into insoluble active particles during the process of purification, which could serve as immobilized bifunctional enzymes. Notably, the immobilized chimera xylanase-lichenase showed a remarkable stability even after 10 reaction cycles, which retained around 56% (lichenase) and 44% (xylanase) of their initial activities, respectively. Moreover, the enhanced thermostability of the immobilized enzymes was also achieved. After incubating at 60 °C for 60 min, the residual activity of the immobilized lichenase was 35%, while the free one was only 24%. Unexpectedly, the free xylanase almost lost its activity when incubated at 55 °C for 60 min, whereas the immobilized xylanase retained 10% of its activity. However, the catalytic efficiency (k cat /K m ) of the free xylanase was 1.7-fold higher than the immobilized one, while the free lichenase was 1.1-fold higher than the immobilized one. This is among the first known reports that two enzymes are purified and immobilized in one-step. This novel strategy is easy to scale up and may meet the demands of biofuel industry. It would have great potentials in other biotechnological fields, such as the multifunctional biomaterials systems. Copyright © 2018 Elsevier Inc. All rights reserved.

Molecular description of the LCST behavior of an elastin-like polypeptide.

PubMed

Li, Nan K; García Quiroz, Felipe; Hall, Carol K; Chilkoti, Ashutosh; Yingling, Yaroslava G

2014-10-13

Elastin-like polypeptides (ELPs) with the repeat sequence of VPGVG are widely used as a model system for investigation of lower critical solution temperature (LCST) transition behavior. In this paper, the effect of temperature on the structure, dynamics and association of (VPGVG)18 in aqueous solution is investigated using atomistic molecular dynamics simulations. Our simulations show that as the temperature increases the ELP backbones undergo gradual conformational changes, which are attributed to the formation of more ordered secondary structures such as β-strands. In addition, increasing temperature changes the hydrophobicity of the ELP by exposure of hydrophobic valine-side chains to the solvent and hiding of proline residues. Based on our simulations, we conclude that the transition behavior of (VPGVG)18 can be attributed to a combination of thermal disruption of the water network that surrounds the polypeptide, reduction of solvent accessible surface area of the polypeptide, and increase in its hydrophobicity. Simulations of the association of two (VPGVG)18 molecules demonstrated that the observed gradual changes in the structural properties of the single polypeptide chain are enough to cause the aggregation of polypeptides above the LCST. These results lead us to propose that the LCST phase behavior of poly(VPGVG) is a collective phenomenon that originates from the correlated gradual changes in single polypeptide structure and the abrupt change in properties of hydration water around the peptide and is a result of a competition between peptide-peptide and peptide-water interactions. This is a computational study of an important intrinsically disordered peptide system that provides an atomic-level description of structural features and interactions that are relevant in the LCST phase behavior.

The Polychromatic Laser Guide Star for tilt measurement: progress report of the demonstrator at Observatoire de Haute Provence

NASA Astrophysics Data System (ADS)

Foy, Renaud; Éric, Pierre; Eysseric, Jérôme; Foy, Françoise; Fusco, Thierry; Girard, Julien; Le Van Suu, Auguste; Perruchot, Sandrine; Richaud, Pierre; Richaud, Yoann; Rondeau, Xavier; Tallon, Michel; Thiébaut, Éric; Boër, Michel

2007-09-01

The Polychromatic Laser Guide Star aims at providing for the tilt measurement from a LGS without any natural guide star. Thus it allows adaptive optics to provide us with a full sky coverage. This is critical in particular to extend adaptive optics to the visible range, where isoplanatism is so small that the probability is negligible to find a natural star to measure the tilt. We report new results obtained within the framework of the Polychromatic LGS programme ELP-OA. Natural stars have been used to mimic the PLGS, in order to check the feasibility of using the difference in the tilt at two wavelengths to derive the tilt itself. We report results from the ATTILA experiment obtained at the 1.52 m telescope at Observatoire de Haute-Provence. Tilts derived from the differential tilts are compared with direct tilt measurements. The accuracy of the measurements is currently ~ 1.5 Airy disk rms at 550 nm. These results prove the feasibility of the Polychromatic Laser Guide Star programme ELP-OA. New algorithms based on inverse problems under development within our programme would lead to smaller error bars by 1 magnitude, as soon as they will run fast enough. We describe the ELP-OA demonstrator which we are setting up at the same telescope, with a special emphasis on the optimization of the excitation process, which definitely has to rely on the two-photon excitation of sodium atoms in the mesosphere. We will describe the implementation at the telescope, including the projector device, the focal instrumentation and the NdYAG pumped dye lasers.

Hydrophobic Hydration of Stimulus-Responsive Polyproteins Measured by Single Molecule Force Spectroscopy

NASA Astrophysics Data System (ADS)

Zauscher, Stefan

2007-03-01

We present a new procedure to reduce and analyze force-extension data obtained by single molecule force spectroscopy (SMFS). This approach allows, for the first time, to infer effects of solvent quality and minor changes in molecular architecture on molecular-elasticity of individual (bio)macromolecules. Specifically, we show how changes in the effective Kuhn segment length can be used to interpret the hydrophobic hydration behavior of elastin-like polypeptides (ELPs).Our results are intriguing as they suggest that SMFS in combination with our analysis procedure can be used to study the subtleties of polypeptide-water interactions on the single molecule level. We also report on the force-induced cis-trans isomerization of prolines, which are repeated every fifth residue in the main chain of ELPs. We present evidence for this mechanism by Monte Carlo simulations of the force-extension curves using an elastically coupled two-state system. Our results suggest that SMFS could be used to assay proline cis-trans isomerization in proteins and may thus have significant potential diagnostic utility.

Trimethylamine N-oxide stabilizes proteins via a distinct mechanism compared with betaine and glycine

PubMed Central

Liao, Yi-Ting; Manson, Anthony C.; DeLyser, Michael R.; Noid, William G.; Cremer, Paul S.

2017-01-01

We report experimental and computational studies investigating the effects of three osmolytes, trimethylamine N-oxide (TMAO), betaine, and glycine, on the hydrophobic collapse of an elastin-like polypeptide (ELP). All three osmolytes stabilize collapsed conformations of the ELP and reduce the lower critical solution temperature (LSCT) linearly with osmolyte concentration. As expected from conventional preferential solvation arguments, betaine and glycine both increase the surface tension at the air–water interface. TMAO, however, reduces the surface tension. Atomically detailed molecular dynamics (MD) simulations suggest that TMAO also slightly accumulates at the polymer–water interface, whereas glycine and betaine are strongly depleted. To investigate alternative mechanisms for osmolyte effects, we performed FTIR experiments that characterized the impact of each cosolvent on the bulk water structure. These experiments showed that TMAO red-shifts the OH stretch of the IR spectrum via a mechanism that was very sensitive to the protonation state of the NO moiety. Glycine also caused a red shift in the OH stretch region, whereas betaine minimally impacted this region. Thus, the effects of osmolytes on the OH spectrum appear uncorrelated with their effects upon hydrophobic collapse. Similarly, MD simulations suggested that TMAO disrupts the water structure to the least extent, whereas glycine exerts the greatest influence on the water structure. These results suggest that TMAO stabilizes collapsed conformations via a mechanism that is distinct from glycine and betaine. In particular, we propose that TMAO stabilizes proteins by acting as a surfactant for the heterogeneous surfaces of folded proteins. PMID:28228526

[Comparative study on microplate and anchor fixation in open-door cervical expansive laminoplasty].

PubMed

Zeng, Yun; Xiong, Min; Yu, Hualong; He, Ning; Wang, Zhiyong; Liu, Zhigang; Han, Heng; Chen, Sen

2011-08-01

To evaluate the effectiveness of microplate fixation in open-door cervical expansive laminoplasty (ELP) by comparing with anchor fixation. Between January 2005 and October 2008, 35 patients with multi-segment cervical spondylotic myelopathy were treated. Of them, 15 patients underwent ELP by microplate fixation (microplate group) and 20 patients underwent ELP by anchor fixation (anchor group). In microplate group, there were 10 males and 5 females with the age of (51.2 +/- 11.5) years; the disease duration ranged from 6 to 60 months (mean, 14 months); and the preoperative Japanese Orthopaedic Association (JOA) score was 7.7 +/- 2.5. In anchor group, there were 13 males and 7 females with the age of (50.7 +/- 10.8) years; the disease duration ranged from 3 to 58 months (mean, 17 months); and the preoperative JOA score was 7.8 +/- 2.9. There was no significant difference in the general data, such as gender, age, and JOA score between 2 groups (P > 0.05). All incisions healed by first intention. Thirty-five cases were followed up 24-68 months (mean, 32 months). The operation time was (113 +/- 24) minutes in anchor group and (111 +/- 27) minutes in microplate group, showing no significant difference (t = 0.231 3, P = 0.818 5). The rate of spinal canal expansion in microplate group (60% +/- 24%) was significantly higher than that in anchor group (40% +/- 18%) (t = 2.820, P = 0.008). The JOA scores of 2 groups at 3 months and 24 months after operation were significantly higher than the preoperative scores (P < 0.01). There was no significant difference in JOA score between 2 groups at 3 months after operation (t = 1.620 5, P = 0.114 6), but the JOA score of microplate group was significantly higher than that of anchor group at 24 months after operation (t = 3.454 3, P = 0.001 5). X-ray film, MRI, and CT scan at 3-6 months after operation displayed that door spindle reached bony fusion. There was no occurrence of "re-close of door" in 2 groups. The rate of complication in microplate group (13.3%, 2/15) was significantly lower than that in anchor group (25.0%, 5/20) (chi2 = 7.160 0, P = 0.008 6). ELP by microplate fixation can achieve the stability quickly after operation, which can help patients to do functional exercises early, and has satisfactory effectiveness and less complications.

Progressive response of large intestinal bacterial community and fermentation to the stepwise decrease of dietary crude protein level in growing pigs.

PubMed

Peng, Yu; Yu, Kaifan; Mu, Chunlong; Hang, Suqin; Che, Lianqiang; Zhu, Weiyun

2017-07-01

The study aimed to determine the effects of reduction of dietary crude protein (CP) level with balanced essential amino acids (EAA) on intestinal bacteria and their metabolites of growing pigs. Forty pigs (initial BW 13.50 ± 0.50 kg, 45 ± 2 days of age) were randomly assigned to four dietary treatments containing CP levels at 20.00% (normal crude protein, NP); 17.16% (medium crude protein, MP); 15.30% (low crude protein, LP); and 13.90% (extremely low crude protein, ELP), respectively. Crystalline AAs were added to meet the EAA requirement of pigs. After 4-week feeding, eight pigs per treatment (n = 8) were randomly selected and slaughtered for sampling of ileal, cecal, and colonic digesta and mucosa. Pigs with moderately reduced CP level had increased bacterial diversity, with the Shannon diversity indices for the colon digesta in the LP group and mucosa in the MP and LP groups significantly (P < 0.05) higher than those in the NP and ELP groups. As the CP level reduces, the Bifidobacterium population were linearly decreased (P < 0.05) both in ileum, cecum, and colon, and the ELP group had the lowest Bifidobacterium population in the cecum and colon, with its value significantly lower than NP and MP groups (P < 0.05). However, the ELP group had the highest population of Escherichia coli in the colon, with its value significantly higher than the LP group (P < 0.05). For bacterial metabolites, as CP level decreased, total short-chain fatty acid (T-SCFA), acetate, and butyrate were linearly increased (linear, P < 0.05) in the ileum, while all SCFAs except formate in the cecum and T-SCFA and acetate in the colon, were linearly decreased (P < 0.05). Reducing CP level led to a linear decrease of microbial crude protein (MCP) in the ileum (P < 0.05) and ammonia in all intestine segments (P < 0.05). The spermidine in cecum and total amines, cadaverine, methylamine, and spermidine in colon were shown a quadratic change (P < 0.05) as dietary CP decreases, with the highest concentration in LP group. These findings suggest that moderate reduction of dietary CP level may benefit large intestinal bacterial community and its fermentation, which was negatively affected by extremely low CP diet.

Using Electronic Portfolios for Second Language Assessment

ERIC Educational Resources Information Center

Cummins, Patricia W.; Davesne, Celine

2009-01-01

Portfolio assessment as developed in Europe presents a learner-empowering alternative to computer-based testing. The authors present the European Language Portfolio (ELP) and its American adaptations, LinguaFolio and the Global Language Portfolio, as tools to be used with the Common European Framework of Reference for languages and the American…

Accurately Predicting Future Reading Difficulty for Bilingual Latino Children at Risk for Language Impairment

ERIC Educational Resources Information Center

Petersen, Douglas B.; Gillam, Ronald B.

2013-01-01

Sixty-three bilingual Latino children who were at risk for language impairment were administered reading-related measures in English and Spanish (letter identification, phonological awareness, rapid automatized naming, and sentence repetition) and descriptive measures including English language proficiency (ELP), language ability (LA),…

Fractionating English Language Proficiency: Policy and Practice in Australian Higher Education

ERIC Educational Resources Information Center

Moore, Paul J.; Harrington, Michael

2016-01-01

Policy-makers and practitioners in Australian higher education increasingly view English language proficiency (ELP) as a tripartite construct consisting of English proficiency for academic study, interpersonal communication, and more recently, the workplace. This paper examines this assumption and identifies the significant implications it has for…

Integrin-mediated targeting of protein polymer nanoparticles carrying a cytostatic macrolide

NASA Astrophysics Data System (ADS)

Shi, Pu

Cytotoxicity, low water solubility, rapid clearance from circulation, and offtarget side-effects are common drawbacks of conventional small-molecule drugs. To overcome these shortcomings, many multifunctional nanocarriers have been proposed to enhance drug delivery. In concept, multifunctional nanoparticles might carry multiple agents, control release rate, biodegrade, and utilize target-mediated drug delivery; however, the design of these particles presents many challenges at the stage of pharmaceutical development. An emerging solution to improve control over these particles is to turn to genetic engineering. Genetically engineered nanocarriers are precisely controlled in size and structure and can provide specific control over sites for chemical attachment of drugs. Genetically engineered drug carriers that assemble nanostructures including nanoparticles and nanofibers can be polymeric or nonpolymeric. This chapter summarizes the recent development of applications in drug and gene delivery utilizing nanostructures of polymeric genetically engineered drug carriers such as elastin-like polypeptides, silk-like polypeptides, and silk-elastin-like protein polymers, and non-polymeric genetically engineered drug carriers such as vault proteins and viral proteins. This chapter explores an alternative encapsulation strategy based on high-specificity avidity between a small molecule drug and its cognate protein target fused to the corona of protein polymer nanoparticles. With the new strategy, the drug associates tightly to the carrier and releases slowly, which may decrease toxicity and promote tumor accumulation via the enhanced permeability and retention effect. To test this hypothesis, the drug Rapamycin (Rapa) was selected for its potent anti-proliferative properties, which give it immunosuppressant and anti-tumor activity. Despite its potency, Rapa has low solubility, low oral bioavailability, and rapid systemic clearance, which make it an excellent candidate for nanoparticulate drug delivery. To explore this approach, genetically engineered diblock copolymers were constructed from elastin-like polypeptides (ELPs) that assemble small nanoparticles. ELPs are protein polymers of the sequence (Val-Pro-Gly-Xaa-Gly)n, where the identity of Xaa and n determine their assembly properties. Initially, a screening assay for model drug encapsulation in ELP nanoparticles was developed, which showed that Rose Bengal and Rapa have high non-specific encapsulation in the core of ELP nanoparticles with a sequence where Xaa = Ile or Phe. While excellent at entrapping these drugs, their release was relatively fast compared to their intended mean residence time in the human body. Having determined that Rapa can be non-specifically entrapped in the core of ELP nanoparticles, FK506 binding protein 12 (FKBP), which is the cognate protein target of Rapa, was genetically fused to the surface of these nanoparticles (FSI) to enhance their avidity towards Rapa. The fusion of FKBP to these nanoparticles slowed the terminal half-life of drug release to 57.8 h. To determine if this class of drug carriers has potential applications in vivo, FSI/Rapa was administered to mice carrying a human breast cancer model (MDA-MB-468). Compared to free drug, FSI encapsulation significantly decreased gross toxicity and enhanced the anti-cancer activity. In conclusion, protein polymer nanoparticles decorated with the cognate receptor of a high potency, low solubility drug (Rapa) efficiently improved drug loading capacity and its release. This approach has applications to the delivery of Rapa and its analogs; furthermore, this strategy has broader applications in the encapsulation, targeting, and release of other potent small molecules. Elastin-like polypeptides (ELPs) are genetically encoded protein polymers that reversibly phase separate in response to stimuli. They respond sharply to small shifts in temperature and form dense microdomains in the living eukaryotic cytosol. This chapter illustrates how to tune the ELP sequence and architecture for either coassembly or sorting of distinct proteins into microdomains within a living cell. Passive tumor targeting utilizing enhanced permeability and retention (EPR) effect has limited efficiency in targeting non-leaky tumors such as MDA-MB-468 breast tumor; however, an RGD tri-peptide decorated micelle nanoparticle can effectively accumulate in tumor site via integrin-mediated active tumor targeting. Different from inefficient and cytotoxic chemical linkage reactions, an elastin-based multi-functional nanocarrier can be assembled by genetic protein fusion and micelle co-assembly technology. The novel drug carrier contains the cognate Rapamycin (Rapa) receptor -- FK506 binding protein (FKBP) as the high-avidity drug binding domain and an RGD peptide as the active tumor targeting domain. Here we show that by co-assembling FKBP and RGD contained protein polymers into mixed micelle nanoparticles, they not only competently targeted endothelial and tumor cells in cell assays, but specifically delivered the drug with a slow release half-life of 38h. It was demonstrated that the active tumor targeting formulation of Rapa more effectively suppressed tumor growth compared to the passive tumor targeting formulation and free drug in tumor regression studies of mouse MDA-MB-468 xenografts. We believe that the exciting results will provide a new tool for the development of next-generation "smart" multi-functional drug carriers. (Abstract shortened by UMI.).

The Challenge of Experiential Learning.

ERIC Educational Resources Information Center

Blower, Deborah F.; Parsons, Nancy A.

The Child Care Services Course (CCSC) at Red River Community College in Manitoba offers an innovative experiential learning component for the first year of a two-year diploma program. To apply for the experiential learning program (ELP), applicants must have 2 years of full-time experience in a children's center or licensed day care home working…

A Potentially Heteroglossic Policy Becomes Monoglossic in Context: An Ethnographic Analysis of Paraguayan Bilingual Education Policy

ERIC Educational Resources Information Center

Mortimer, Katherine S.

2016-01-01

Ethnographic and discursive approaches to educational language policy (ELP) that explore how policy is appropriated in context are important for understanding policy success/failure in meeting goals of educational equity for language-minoritized students. This study describes how Paraguayan national policy for universal bilingual education…

The ELP through Time: Background Motivation, Growing Experience, Current Beliefs

ERIC Educational Resources Information Center

Argondizzo, Carmen; Sasso, Maria I.

2016-01-01

This article offers an overview of research strategies currently in use at the Language Centre of the University of Calabria and aimed at observing university students' learning habits when they are asked to use the European Language Portfolio during language courses. We present evidence of how experimental groups of students belonging to…

A Framework for International Student Participation in Postsecondary U.S. English Language Programs

ERIC Educational Resources Information Center

Colón, Valeriana

2016-01-01

Postsecondary English language education is a growing field in the United States. While there has been considerable research on international student mobility in higher education, there is limited research on the population's participation in U.S. English language programs (ELPs). This study examined literature in related fields to create a…

The Effects of English Language Proficiency and Curricular Pathways: Latina/os' Mathematics Achievement in Secondary Schools

ERIC Educational Resources Information Center

Mosqueda, Eduardo; Maldonado, Saul I.

2013-01-01

This study analyzes nationally-representative quantitative data from the first (2002) and second (2004) waves of the Educational Longitudinal Study to examine the relationship between Latina/o secondary school students' degree of English-language proficiency (ELP), mathematics course-taking measures, and 12th grade mathematics achievement.…

An Application of the EPOSTL: Focus on Culture

ERIC Educational Resources Information Center

Arikan, Arda; Zorba, M. Galip

2016-01-01

Although Turkey is not a member state of the European Union (EU), educational and curricular documents produced by the EU such like the Common European Framework of Reference for Languages (CEFR), the European Language Portfolio (the ELP) and the European Portfolio for Student Teachers of Languages (EPOSTL) have been examined by Turkish…

Fostering Self-Regulated Learning through the European Language Portfolio: An Embedded Mixed Methods Study

ERIC Educational Resources Information Center

Ziegler, Nicholas Allan

2014-01-01

The European Language Portfolio (ELP) is an alternative assessment used in foreign language classes throughout Europe to support and record language learning. Directly linked to the Common European Framework of Reference for Languages (Council of Europe, 2001) proficiency guidelines, it is designed to achieve an ambitious dual goal: document…

Experiences with Autonomy: Learners' Voices on Language Learning

ERIC Educational Resources Information Center

Kristmanson, Paula; Lafargue, Chantal; Culligan, Karla

2013-01-01

This article focuses on the experiences of Grade 12 students using a language portfolio based on the principles and guidelines of the European Language Portfolio (ELP) in their second language classes in a large urban high school. As part of a larger action-research project, focus group interviews were conducted to gather data related to…

Educational Leadership Effectiveness: A Rasch Analysis

ERIC Educational Resources Information Center

Sinnema, Claire; Ludlow, Larry; Robinson, Viviane

2016-01-01

Purpose: The purposes of this paper are, first, to establish the psychometric properties of the ELP tool, and, second, to test, using a Rasch item response theory analysis, the hypothesized progression of challenge presented by the items included in the tool. Design/ Methodology/ Approach: Data were collected at two time points through a survey of…

Development of a portable Linux-based ECG measurement and monitoring system.

PubMed

Tan, Tan-Hsu; Chang, Ching-Su; Huang, Yung-Fa; Chen, Yung-Fu; Lee, Cheng

2011-08-01

This work presents a portable Linux-based electrocardiogram (ECG) signals measurement and monitoring system. The proposed system consists of an ECG front end and an embedded Linux platform (ELP). The ECG front end digitizes 12-lead ECG signals acquired from electrodes and then delivers them to the ELP via a universal serial bus (USB) interface for storage, signal processing, and graphic display. The proposed system can be installed anywhere (e.g., offices, homes, healthcare centers and ambulances) to allow people to self-monitor their health conditions at any time. The proposed system also enables remote diagnosis via Internet. Additionally, the system has a 7-in. interactive TFT-LCD touch screen that enables users to execute various functions, such as scaling a single-lead or multiple-lead ECG waveforms. The effectiveness of the proposed system was verified by using a commercial 12-lead ECG signal simulator and in vivo experiments. In addition to its portability, the proposed system is license-free as Linux, an open-source code, is utilized during software development. The cost-effectiveness of the system significantly enhances its practical application for personal healthcare.

Food grade microemulsion systems: Sunflower oil/castor oil derivative-ethanol/water. Rheological and physicochemical analysis.

PubMed

Mori Cortés, Noelia; Lorenzo, Gabriel; Califano, Alicia N

2018-05-01

Microemulsions are thermodynamically stable systems that have attracted considerable attention in the food industry as delivery systems for many hydrophobic nutrients. These spontaneous systems are highly dependent on ingredients and composition. In this work phase diagrams were constructed using two surfactants (Kolliphor RH40 and ELP), water, sunflower oil, and ethanol as cosurfactant, evaluating their physicochemical properties. Stability of the systems was studied at 25 and 60 °C, monitoring turbidity at 550 nm for over a month to identify the microemulsion region. Conductivity was measured to classify between water-in-oil and oil-in-water microemulsions. The phase diagram constructed with Kolliphor RH40 exhibited a larger microemulsion area than that formulated with Kolliphor ELP. All formulations showed a monomodal droplet size distribution with low polydispersity index (<0.30) and a mean droplet size below 20 nm. Systems with higher water content presented a Newtonian behavior; increasing the dispersed phase content produced a weak gel-like structure with pseudoplastic behavior under flow conditions that was satisfactorily modeled to obtain structural parameters. Copyright © 2018 Elsevier Ltd. All rights reserved.

tRNA tKUUU, tQUUG, and tEUUC wobble position modifications fine-tune protein translation by promoting ribosome A-site binding.

PubMed

Rezgui, Vanessa Anissa Nathalie; Tyagi, Kshitiz; Ranjan, Namit; Konevega, Andrey L; Mittelstaet, Joerg; Rodnina, Marina V; Peter, Matthias; Pedrioli, Patrick G A

2013-07-23

tRNA modifications are crucial to ensure translation efficiency and fidelity. In eukaryotes, the URM1 and ELP pathways increase cellular resistance to various stress conditions, such as nutrient starvation and oxidative agents, by promoting thiolation and methoxycarbonylmethylation, respectively, of the wobble uridine of cytoplasmic (tK(UUU)), (tQ(UUG)), and (tE(UUC)). Although in vitro experiments have implicated these tRNA modifications in modulating wobbling capacity and translation efficiency, their exact in vivo biological roles remain largely unexplored. Using a combination of quantitative proteomics and codon-specific translation reporters, we find that translation of a specific gene subset enriched for AAA, CAA, and GAA codons is impaired in the absence of URM1- and ELP-dependent tRNA modifications. Moreover, in vitro experiments using native tRNAs demonstrate that both modifications enhance binding of tK(UUU) to the ribosomal A-site. Taken together, our data suggest that tRNA thiolation and methoxycarbonylmethylation regulate translation of genes with specific codon content.

An injectable elastin-based gene delivery platform for dose-dependent modulation of angiogenesis and inflammation for critical limb ischemia.

PubMed

Dash, Biraja C; Thomas, Dilip; Monaghan, Michael; Carroll, Oliver; Chen, Xizhe; Woodhouse, Kimberly; O'Brien, Timothy; Pandit, Abhay

2015-10-01

Critical limb ischemia is a major clinical problem. Despite rigorous treatment regimes, there has been only modest success in reducing the rate of amputations in affected patients. Reduced level of blood flow and enhanced inflammation are the two major pathophysiological changes that occur in the ischemic tissue. The objective of this study was to develop a controlled dual gene delivery system capable of delivering therapeutic plasmid eNOS and IL-10 in a temporal manner. In order to deliver multiple therapeutic genes, an elastin-like polypeptide (ELP) based injectable system was designed. The injectable system was comprised of hollow spheres and an in situ-forming gel scaffold of elastin-like polypeptide capable of carrying gene complexes, with an extended manner release profile. In addition, the ELP based injectable system was used to deliver human eNOS and IL-10 therapeutic genes in vivo. A subcutaneous dose response study showed enhanced blood vessel density in the treatment groups of eNOS (20 μg) and IL-10 (10 μg)/eNOS (20 μg) and reduced inflammation with IL-10 (10 μg) alone. Next, we carried out a hind-limb ischemia model comparing the efficacy of the following interventions; Saline; IL-10, eNOS and IL-10/eNOS. The selected dose of eNOS, exhibited enhanced angiogenesis. IL-10 treatment groups showed reduction in the level of inflammatory cells. Furthermore, we demonstrated that eNOS up-regulated major proangiogenic growth factors such as vascular endothelial growth factors, platelet derived growth factor B, and fibroblast growth factor 1, which may explain the mechanism of this approach. These factors help in formation of a stable vascular network. Thus, ELP injectable system mediating non-viral delivery of human IL10-eNOS is a promising therapy towards treating limb ischemia. Copyright © 2015 Elsevier Ltd. All rights reserved.

The familial dysautonomia disease gene IKBKAP is required in the developing and adult mouse central nervous system

PubMed Central

Chaverra, Marta; George, Lynn; Thorne, Julian; Grindeland, Andrea; Ueki, Yumi; Eiger, Steven; Cusick, Cassie; Babcock, A. Michael; Carlson, George A.

2017-01-01

ABSTRACT Hereditary sensory and autonomic neuropathies (HSANs) are a genetically and clinically diverse group of disorders defined by peripheral nervous system (PNS) dysfunction. HSAN type III, known as familial dysautonomia (FD), results from a single base mutation in the gene IKBKAP that encodes a scaffolding unit (ELP1) for a multi-subunit complex known as Elongator. Since mutations in other Elongator subunits (ELP2 to ELP4) are associated with central nervous system (CNS) disorders, the goal of this study was to investigate a potential requirement for Ikbkap in the CNS of mice. The sensory and autonomic pathophysiology of FD is fatal, with the majority of patients dying by age 40. While signs and pathology of FD have been noted in the CNS, the clinical and research focus has been on the sensory and autonomic dysfunction, and no genetic model studies have investigated the requirement for Ikbkap in the CNS. Here, we report, using a novel mouse line in which Ikbkap is deleted solely in the nervous system, that not only is Ikbkap widely expressed in the embryonic and adult CNS, but its deletion perturbs both the development of cortical neurons and their survival in adulthood. Primary cilia in embryonic cortical apical progenitors and motile cilia in adult ependymal cells are reduced in number and disorganized. Furthermore, we report that, in the adult CNS, both autonomic and non-autonomic neuronal populations require Ikbkap for survival, including spinal motor and cortical neurons. In addition, the mice developed kyphoscoliosis, an FD hallmark, indicating its neuropathic etiology. Ultimately, these perturbations manifest in a developmental and progressive neurodegenerative condition that includes impairments in learning and memory. Collectively, these data reveal an essential function for Ikbkap that extends beyond the peripheral nervous system to CNS development and function. With the identification of discrete CNS cell types and structures that depend on Ikbkap, novel strategies to thwart the progressive demise of CNS neurons in FD can be developed. PMID:28167615

A bioinspired elastin-based protein for a cytocompatible underwater adhesive.

PubMed

Brennan, M Jane; Kilbride, Bridget F; Wilker, Jonathan J; Liu, Julie C

2017-04-01

The development of adhesives that can be applied and create strong bonds underwater is a significant challenge for materials engineering. When the adhesive is intended for biomedical applications, further criteria, such as biocompatibility, must be met. Current biomedical adhesive technologies do not meet these needs. In response, we designed a bioinspired protein system that shows promise to achieve biocompatible underwater adhesion coupled with environmentally responsive behavior that is "smart" - that is, it can be tuned to suit a specific application. The material, ELY 16 , is constructed from an elastin-like polypeptide (ELP) that can be produced in high yields from Escherichia coli and can coacervate in response to environmental factors such as temperature, pH, and salinity. To confer wet adhesion, we utilized design principles from marine organisms such as mussels and sandcastle worms. When expressed, ELY 16 is rich in tyrosine. Upon modification with the tyrosinase enzyme to form mELY 16 , the tyrosine residues are converted to 3,4-dihydroxyphenylalanine (DOPA). Both ELY 16 and mELY 16 exhibit cytocompatibility and significant dry adhesion strength (>2 MPa). Modification with DOPA increases protein adsorption to glass and provides moderate adhesion strength (∼240 kPa) in a highly humid environment. Furthermore, this ELP exhibits a tunable phase transition behavior that can be formulated to coacervate in physiological conditions and provides a convenient mechanism for application underwater. Finally, mELY 16 possesses significantly higher adhesion strength in dry, humid, and underwater environments compared with a commercially available fibrin sealant. To our knowledge, mELY 16 provides the strongest bonds of any rationally designed protein when used completely underwater, and its high yields make it more viable for commercial application compared to natural adhesive proteins. In conclusion, this ELP shows great potential to be a new "smart" underwater adhesive. Copyright © 2017 Elsevier Ltd. All rights reserved.

The effect of dimethylsulfoxide on the water transport response of rat hepatocytes during freezing.

PubMed

Smith, D J; Schulte, M; Bischof, J C

1998-10-01

Successful improvement of cryopreservation protocols for cells in suspension requires knowledge of how such cells respond to the biophysical stresses of freezing (intracellular ice formation, water transport) while in the presence of a cryoprotective agent (CPA). This work investigates the biophysical water transport response in a clinically important cell type--isolated hepatocytes--during freezing in the presence of dimethylsulfoxide (DMSO). Sprague-Dawley rat liver hepatocytes were frozen in Williams E media supplemented with 0, 1, and 2 M DMSO, at rates of 5, 10, and 50 degrees C/min. The water transport was measured by cell volumetric changes as assessed by cryomicroscopy and image analysis. Assuming that water is the only species transported under these conditions, a water transport model of the form dV/dT = f(Lpg([CPA]), ELp([CPA]), T(t)) was curve-fit to the experimental data to obtain the biophysical parameters of water transport--the reference hydraulic permeability (Lpg) and activation energy of water transport (ELp)--for each DMSO concentration. These parameters were estimated two ways: (1) by curve-fitting the model to the average volume of the pooled cell data, and (2) by curve-fitting individual cell volume data and averaging the resulting parameters. The experimental data showed that less dehydration occurs during freezing at a given rate in the presence of DMSO at temperatures between 0 and -10 degrees C. However, dehydration was able to continue at lower temperatures (< -10 degrees C) in the presence of DMSO. The values of Lpg and ELp obtained using the individual cell volume data both decreased from their non-CPA values--4.33 x 10(-13) m3/N-s (2.69 microns/min-atm) and 317 kJ/mol (75.9 kcal/mol), respectively--to 0.873 x 10(-13) m3/N-s (0.542 micron/min-atm) and 137 kJ/mol (32.8 kcal/mol), respectively, in 1 M DMSO and 0.715 x 10(-13) m3/N-s (0.444 micron/min-atm) and 107 kJ/mol (25.7 kcal/mol), respectively, in 2 M DMSO. The trends in the pooled volume values for Lpg and ELp were very similar, but the overall fit was considered worse than for the individual volume parameters. A unique way of presenting the curve-fitting results supports a clear trend of reduction of both biophysical parameters in the presence of DMSO, and no clear trend in cooling rate dependence of the biophysical parameters. In addition, these results suggest that close proximity of the experimental cell volume data to the equilibrium volume curve may significantly reduce the efficiency of the curve-fitting process.

Autonomy and Self-Assessment of Individual Learning Styles Using the European Language Portfolio (ELP)

ERIC Educational Resources Information Center

Peréz Cavana, María Luisa

2012-01-01

Interest in learning styles has produced innumerable studies over the last three decades. However, the application of knowledge of learning styles in education is a controversial matter, particularly in instruction, where there is less evidence of the usefulness of learning styles tests. This article is concerned with the role of reflection in…

Accommodations for English Language Learners with Disabilities in Federally-Mandated Statewide English Language Proficiency Assessments

ERIC Educational Resources Information Center

Kuti, Laura M.

2011-01-01

The purpose of this research study was to conduct exploratory research to examine federally-mandated annual English language proficiency (ELP) assessment of English language learners (ELLs) and their use of accommodations on the assessment. First the researcher provides a review of the literature regarding accommodations provided for ELLs,…

From the Learning Diary to the ELP: An E-Portfolio for Autonomous Language Learning

ERIC Educational Resources Information Center

Bertolotti, Greta; Beseghi, Micol

2016-01-01

In 2011 the Language Centre of the University of Parma introduced a self-study programme aimed at creating an autonomy-inspired language learning environment. Students are actively engaged in the management of their own learning and co-directed by advisors and teachers in the phases of planning, monitoring and assessment. Reflective diary writing…

Promoting Student Autonomy through the Use of the European Language Portfolio

ERIC Educational Resources Information Center

Gonzalez, Jesus Angel

2009-01-01

The European Language Portfolio (ELP) is a document launched by the Council of Europe in 2001 which consists of three sections: the Passport, the Language Biography, and the Dossier. It has two complementary functions: a pedagogic function (helping students to reflect on their learning and objectives) and a reporting function (providing a record…

"Putting Music On": Everyday Leisure Activities, Choice-Making and Person-Centred Planning in a Supported Living Scheme

ERIC Educational Resources Information Center

Hassan, Nedim

2017-01-01

Background: Person-centred planning, which commonly becomes formalised within services for people with learning disabilities through an Essential Lifestyle Plan (ELP), was intended to help place the choices of individuals at the forefront of service provision. However, beyond UK government policy rhetoric, scholars have raised issues regarding the…

The Achievement Progress of English Learner Students in Nevada. REL 2016-154

ERIC Educational Resources Information Center

Haas, Eric; Huang, Min; Tran, Loan; Yu, Airong

2016-01-01

The purpose of this study was to examine the cumulative progress of English learner students in Nevada in English language proficiency (ELP) and in academic content knowledge in both reading and mathematics. This study identified students in grades kindergarten, 3, and 6 who were designated as English learner students in 2006/07 and examined their…

Practitioners' Evaluation on the Procedural Aspects of an English Language Portfolio

ERIC Educational Resources Information Center

Karababa, Canan; Suzer, Sezgi Sarac

2010-01-01

This study aims to share English language practitioners' reflections on the adaptation of teaching methods and materials during the pre-validation period of the European Language Portfolio (ELP). In order to achieve this aim, a high school was selected as the scope of research, since it has started to adapt the English language teaching…

English Learner "Curricular Streams" in Four Middle Schools: Triage in the Trenches

ERIC Educational Resources Information Center

Estrada, Peggy

2014-01-01

Little is known about the curricular experiences schools provide English learner students (ELs) to meet the dual goals of attaining English language proficiency (ELP) and grade-level achievement. I introduce the concept of "Curricular Streams" to provide a more nuanced comparative analysis of four urban middle schools, focusing on: (a)…

Thermally Targeted Delivery of a c-Myc Inhibitory Peptide In Vivo Using Elastin-like Polypeptide

DTIC Science & Technology

2009-10-01

cytoplasm. Also, in a subset of cells, Bac-ELP1⁎-H1 showed very bright nuclear staining exclusive of nucleoli (Fig. 5, lower right, arrows). 3.6. Time...localization was very bright relative to the amount of polypeptide in the cytoplasm, and it appeared to be nucleoplasmic and excluded from nucleoli . The

Creating a Next-Generation System of K-12 English Learner Language Proficiency Assessments. Research Report. ETS RR-16-06

ERIC Educational Resources Information Center

Hauck, Maurice Cogan; Wolf, Mikyung Kim; Mislevy, Robert

2016-01-01

This paper is the first in a series from Educational Testing Service (ETS) concerning English language proficiency (ELP) assessments for K-12 English learners (ELs). The goal of this paper, and the series, is to present research-based ideas, principles, and recommendations for consideration by those who are conceptualizing, developing, and…

The Achievement Progress of English Learner Students in Utah. REL 2016-155

ERIC Educational Resources Information Center

Haas, Eric; Huang, Min; Tran, Loan; Yu, Airong

2016-01-01

The purpose of this study was to examine the cumulative progress of English learner students in Utah in English language proficiency (ELP) and in academic content knowledge in both English language arts (ELA) and mathematics. This study identified students in grades kindergarten, 3, and 6 who were designated as English learner students in 2006/07…

Implementing Project Based Survey Research Skills to Grade Six ELP Students with "The Survey Toolkit" and "TinkerPlots"[R

ERIC Educational Resources Information Center

Walsh, Thomas, Jr.

2011-01-01

"Survey Toolkit Collecting Information, Analyzing Data and Writing Reports" (Walsh, 2009a) is discussed as a survey research curriculum used by the author's sixth grade students. The report describes the implementation of "The Survey Toolkit" curriculum and "TinkerPlots"[R] software to provide instruction to students learning a project based…

Leading an Independent School Today Means Leading Change

ERIC Educational Resources Information Center

Walpole, Andrew Robert Nicholas

2014-01-01

This Executive Leadership Portfolio (ELP) is the story of my journey through the Ed.D. program that begins with me focusing my work on trying to reverse my school's attrition woes, then changes course as I try to make my project reverse my own failed head of school candidacies, and ends with me discovering that, along the way, I had learned to…

Toward a "Common Definition of English Learner": Guidance for States and State Assessment Consortia in Defining and Addressing Policy and Technical Issues and Options

ERIC Educational Resources Information Center

Linquanti, Robert; Cook, H. Gary

2013-01-01

States participating in the four federally-funded assessment consortia are required to establish a "common definition of English Learner." This includes the two Race to the Top academic assessment consortia and the two Enhanced Assessment Grant English language proficiency (ELP) assessment consortia. This paper provides guidance that…

IL-17 Plays a Role in Respiratory Syncytial Virus-induced Lung Inflammation and Emphysema in Elastase and LPS-injured Mice.

PubMed

Mebratu, Yohannes A; Tesfaigzi, Yohannes

2018-06-01

Respiratory syncytial virus (RSV) is associated with enhanced progression of chronic obstructive pulmonary disease (COPD) and COPD exacerbations. However, little is known about the role of IL-17 in RSV-induced lung injury. We first investigated the role of RSV infection in enhancing mucous cell hyperplasia (MCH) and airspace enlargement in the lungs of mice injured with elastase and LPS (E/LPS). Mice injured with E/LPS had an enhanced and prolonged neutrophilic response to RSV that was associated with decreased levels of type I IFN and increased levels of IL-17, IL-23, CXCL-1, granulocyte colony stimulating factor (GCSF), CXCL-5, and matrix metalloproteinase (MMP)-9. In addition, extent of MCH and mean weighted alveolar space were increased significantly in the lungs of E/LPS-injured mice infected with RSV compared with E/LPS-only or RSV-only controls. Interestingly, immunodepletion of IL-17 before viral infection diminished the RSV-driven MCH and airspace enlargement in the E/LPS-injured animals, suggesting that IL-17 may be a therapeutic target for MCH and airspace enlargement when enhanced by RSV infection.

Recognition of chromatin by the plant alkaloid, ellipticine as a dual binder

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

Banerjee, Amrita; Sanyal, Sulagna; Majumder, Parijat

Recognition of core histone components of chromatin along with chromosomal DNA by a class of small molecule modulators is worth examining to evaluate their intracellular mode of action. A plant alkaloid ellipticine (ELP) which is a putative anticancer agent has so far been reported to function via DNA intercalation, association with topoisomerase II and binding to telomere region. However, its effect upon the potential intracellular target, chromatin is hitherto unreported. Here we have characterized the biomolecular recognition between ELP and different hierarchical levels of chromatin. The significant result is that in addition to DNA, it binds to core histone(s) andmore » can be categorized as a ‘dual binder’. As a sequel to binding with histone(s) and core octamer, it alters post-translational histone acetylation marks. We have further demonstrated that it has the potential to modulate gene expression thereby regulating several key biological processes such as nuclear organization, transcription, translation and histone modifications. - Highlights: • Ellipticine acts a dual binder binding to both DNA and core histone(s). • It induces structural perturbations in chromatin, chromatosome and histone octamer. • It alters histones acetylation and affects global gene expression.« less

Astrometry of the Orcus/Vanth occultation on UT 7 March 2017

NASA Astrophysics Data System (ADS)

Bosh, Amanda S.; Zuluaga, Carlos; Levine, Stephen; Sickafoose, Amanda A.; Genade, Anja; Schindler, Karsten; Lister, Tim; Person, Michael J.

2017-10-01

On UT 7 March 2017, Orcus was predicted to occult a star with m=14.3. Observations were made at five observatories: the 0.6-m Astronomical Telescope of the University of Stuttgart (ATUS) at Sierra Remote Observatories (SRO), California; Las Cumbres Observatory’s 1-m telescope (ELP) at McDonald Observatory, Fort Davis, Texas; NASA’s 3-m InfraRed Telescope Facility (IRTF) on Mauna Kea, Hawaii; the 4.1-m Southern Astrophysical Research telescope (SOAR) on Cerro Pachón, Chile; and the 0.6-m Southeastern Association for Research in Astronomy telescope (SARA-CT) at Cerro Tololo, Chile. While observations at all sites were successful, only two—ELP and IRTF—observed solid-body occultation signatures. We will discuss the various predictions for this event and the reasons for the differences among them, including an offset of 130 mas for the star position from the position in the Gaia catalog. The sum of the positive and negative detections place constraints on the geometry of the Orcus/Vanth system, and we present our astrometric results for the geometric solution for this occultation. The implications of the light curve analyses are presented by Sickafoose et al., this conference.

Genome-wide association analysis identifies three new susceptibility loci for childhood body mass index

PubMed Central

Felix, Janine F.; Bradfield, Jonathan P.; Monnereau, Claire; van der Valk, Ralf J.P.; Stergiakouli, Evie; Chesi, Alessandra; Gaillard, Romy; Feenstra, Bjarke; Thiering, Elisabeth; Kreiner-Møller, Eskil; Mahajan, Anubha; Pitkänen, Niina; Joro, Raimo; Cavadino, Alana; Huikari, Ville; Franks, Steve; Groen-Blokhuis, Maria M.; Cousminer, Diana L.; Marsh, Julie A.; Lehtimäki, Terho; Curtin, John A.; Vioque, Jesus; Ahluwalia, Tarunveer S.; Myhre, Ronny; Price, Thomas S.; Vilor-Tejedor, Natalia; Yengo, Loïc; Grarup, Niels; Ntalla, Ioanna; Ang, Wei; Atalay, Mustafa; Bisgaard, Hans; Blakemore, Alexandra I.; Bonnefond, Amelie; Carstensen, Lisbeth; Eriksson, Johan; Flexeder, Claudia; Franke, Lude; Geller, Frank; Geserick, Mandy; Hartikainen, Anna-Liisa; Haworth, Claire M.A.; Hirschhorn, Joel N.; Hofman, Albert; Holm, Jens-Christian; Horikoshi, Momoko; Hottenga, Jouke Jan; Huang, Jinyan; Kadarmideen, Haja N.; Kähönen, Mika; Kiess, Wieland; Lakka, Hanna-Maaria; Lakka, Timo A.; Lewin, Alexandra M.; Liang, Liming; Lyytikäinen, Leo-Pekka; Ma, Baoshan; Magnus, Per; McCormack, Shana E.; McMahon, George; Mentch, Frank D.; Middeldorp, Christel M.; Murray, Clare S.; Pahkala, Katja; Pers, Tune H.; Pfäffle, Roland; Postma, Dirkje S.; Power, Christine; Simpson, Angela; Sengpiel, Verena; Tiesler, Carla M. T.; Torrent, Maties; Uitterlinden, André G.; van Meurs, Joyce B.; Vinding, Rebecca; Waage, Johannes; Wardle, Jane; Zeggini, Eleftheria; Zemel, Babette S.; Dedoussis, George V.; Pedersen, Oluf; Froguel, Philippe; Sunyer, Jordi; Plomin, Robert; Jacobsson, Bo; Hansen, Torben; Gonzalez, Juan R.; Custovic, Adnan; Raitakari, Olli T.; Pennell, Craig E.; Widén, Elisabeth; Boomsma, Dorret I.; Koppelman, Gerard H.; Sebert, Sylvain; Järvelin, Marjo-Riitta; Hyppönen, Elina; McCarthy, Mark I.; Lindi, Virpi; Harri, Niinikoski; Körner, Antje; Bønnelykke, Klaus; Heinrich, Joachim; Melbye, Mads; Rivadeneira, Fernando; Hakonarson, Hakon; Ring, Susan M.; Smith, George Davey; Sørensen, Thorkild I.A.; Timpson, Nicholas J.; Grant, Struan F.A.; Jaddoe, Vincent W.V.

2016-01-01

A large number of genetic loci are associated with adult body mass index. However, the genetics of childhood body mass index are largely unknown. We performed a meta-analysis of genome-wide association studies of childhood body mass index, using sex- and age-adjusted standard deviation scores. We included 35 668 children from 20 studies in the discovery phase and 11 873 children from 13 studies in the replication phase. In total, 15 loci reached genome-wide significance (P-value < 5 × 10−8) in the joint discovery and replication analysis, of which 12 are previously identified loci in or close to ADCY3, GNPDA2, TMEM18, SEC16B, FAIM2, FTO, TFAP2B, TNNI3K, MC4R, GPR61, LMX1B and OLFM4 associated with adult body mass index or childhood obesity. We identified three novel loci: rs13253111 near ELP3, rs8092503 near RAB27B and rs13387838 near ADAM23. Per additional risk allele, body mass index increased 0.04 Standard Deviation Score (SDS) [Standard Error (SE) 0.007], 0.05 SDS (SE 0.008) and 0.14 SDS (SE 0.025), for rs13253111, rs8092503 and rs13387838, respectively. A genetic risk score combining all 15 SNPs showed that each additional average risk allele was associated with a 0.073 SDS (SE 0.011, P-value = 3.12 × 10−10) increase in childhood body mass index in a population of 1955 children. This risk score explained 2% of the variance in childhood body mass index. This study highlights the shared genetic background between childhood and adult body mass index and adds three novel loci. These loci likely represent age-related differences in strength of the associations with body mass index. PMID:26604143

Genome-wide association analysis identifies three new susceptibility loci for childhood body mass index.

PubMed

Felix, Janine F; Bradfield, Jonathan P; Monnereau, Claire; van der Valk, Ralf J P; Stergiakouli, Evie; Chesi, Alessandra; Gaillard, Romy; Feenstra, Bjarke; Thiering, Elisabeth; Kreiner-Møller, Eskil; Mahajan, Anubha; Pitkänen, Niina; Joro, Raimo; Cavadino, Alana; Huikari, Ville; Franks, Steve; Groen-Blokhuis, Maria M; Cousminer, Diana L; Marsh, Julie A; Lehtimäki, Terho; Curtin, John A; Vioque, Jesus; Ahluwalia, Tarunveer S; Myhre, Ronny; Price, Thomas S; Vilor-Tejedor, Natalia; Yengo, Loïc; Grarup, Niels; Ntalla, Ioanna; Ang, Wei; Atalay, Mustafa; Bisgaard, Hans; Blakemore, Alexandra I; Bonnefond, Amelie; Carstensen, Lisbeth; Eriksson, Johan; Flexeder, Claudia; Franke, Lude; Geller, Frank; Geserick, Mandy; Hartikainen, Anna-Liisa; Haworth, Claire M A; Hirschhorn, Joel N; Hofman, Albert; Holm, Jens-Christian; Horikoshi, Momoko; Hottenga, Jouke Jan; Huang, Jinyan; Kadarmideen, Haja N; Kähönen, Mika; Kiess, Wieland; Lakka, Hanna-Maaria; Lakka, Timo A; Lewin, Alexandra M; Liang, Liming; Lyytikäinen, Leo-Pekka; Ma, Baoshan; Magnus, Per; McCormack, Shana E; McMahon, George; Mentch, Frank D; Middeldorp, Christel M; Murray, Clare S; Pahkala, Katja; Pers, Tune H; Pfäffle, Roland; Postma, Dirkje S; Power, Christine; Simpson, Angela; Sengpiel, Verena; Tiesler, Carla M T; Torrent, Maties; Uitterlinden, André G; van Meurs, Joyce B; Vinding, Rebecca; Waage, Johannes; Wardle, Jane; Zeggini, Eleftheria; Zemel, Babette S; Dedoussis, George V; Pedersen, Oluf; Froguel, Philippe; Sunyer, Jordi; Plomin, Robert; Jacobsson, Bo; Hansen, Torben; Gonzalez, Juan R; Custovic, Adnan; Raitakari, Olli T; Pennell, Craig E; Widén, Elisabeth; Boomsma, Dorret I; Koppelman, Gerard H; Sebert, Sylvain; Järvelin, Marjo-Riitta; Hyppönen, Elina; McCarthy, Mark I; Lindi, Virpi; Harri, Niinikoski; Körner, Antje; Bønnelykke, Klaus; Heinrich, Joachim; Melbye, Mads; Rivadeneira, Fernando; Hakonarson, Hakon; Ring, Susan M; Smith, George Davey; Sørensen, Thorkild I A; Timpson, Nicholas J; Grant, Struan F A; Jaddoe, Vincent W V

2016-01-15

A large number of genetic loci are associated with adult body mass index. However, the genetics of childhood body mass index are largely unknown. We performed a meta-analysis of genome-wide association studies of childhood body mass index, using sex- and age-adjusted standard deviation scores. We included 35 668 children from 20 studies in the discovery phase and 11 873 children from 13 studies in the replication phase. In total, 15 loci reached genome-wide significance (P-value < 5 × 10(-8)) in the joint discovery and replication analysis, of which 12 are previously identified loci in or close to ADCY3, GNPDA2, TMEM18, SEC16B, FAIM2, FTO, TFAP2B, TNNI3K, MC4R, GPR61, LMX1B and OLFM4 associated with adult body mass index or childhood obesity. We identified three novel loci: rs13253111 near ELP3, rs8092503 near RAB27B and rs13387838 near ADAM23. Per additional risk allele, body mass index increased 0.04 Standard Deviation Score (SDS) [Standard Error (SE) 0.007], 0.05 SDS (SE 0.008) and 0.14 SDS (SE 0.025), for rs13253111, rs8092503 and rs13387838, respectively. A genetic risk score combining all 15 SNPs showed that each additional average risk allele was associated with a 0.073 SDS (SE 0.011, P-value = 3.12 × 10(-10)) increase in childhood body mass index in a population of 1955 children. This risk score explained 2% of the variance in childhood body mass index. This study highlights the shared genetic background between childhood and adult body mass index and adds three novel loci. These loci likely represent age-related differences in strength of the associations with body mass index. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

The Biennial Report to Congress on the Implementation of the Title III State Formula Grant Program: School Years 2004-06

ERIC Educational Resources Information Center

Office of English Language Acquisition, US Department of Education, 2008

2008-01-01

The biennial report to Congress on the implementation of the Title III state formula grant program provides a snapshot of the status of the U.S. Department of Education's efforts to hold states accountable for ensuring that all limited English proficient (LEP) students attain English language proficiency (ELP) and are achieving in the content…

Taskmaster: A Prototype Graphical User-Interface to a Schedule Optimization Model

DTIC Science & Technology

1990-03-01

34Assimilation Paradox" .. ........................... 42 c. O nline H elp ......................................... 43 C. DIRECT MANIPULATION...advantage of these hardware capabilit’es. As a result of these changes, new markets are developing for a variety of computer apTIications that were once...the 1989 meeting of ORSATIMS. In that paper the authors make the following observation. 8 With traditionally developed systems, the potential market

Health-related quality of life and care satisfaction outcomes: Informing psychosocial oncology care among Latina and African-American young breast cancer survivors.

PubMed

Ashing, Kimlin Tam; George, Marshalee; Jones, Veronica

2018-04-01

When breast cancer occurs in young women, the medical, physical, psychosocial, and overall impacts can be more severe warranting targeted medical and psychosocial oncology care. Yet, despite their risk for poorer survival and survivorship outcomes, little research has focused on this group with critical gaps concerning ethnic minorities who are particularly medically vulnerable. Therefore, this preliminary study examined demographic characteristics and patient centered outcomes, ie, health-related quality of life (HRQOL), quality care satisfaction to inform targeted psychosocial oncology care among African-American and Latinas young breast cancer survivors (YBCS). A total of 116 African-American and Latina YBCS aged ≥21 to 50 years were recruited from cancer registries and community agencies. Based on prior research and the literature, Latinas were categorized into English language proficient (ELP) and Spanish language proficient (SLP) based on their choice of language to conduct the study including completion of the measures. SLP Latinas reported lower educational attainment and income (P < 0.001) and were more likely to report having a mastectomy (P < 0.01) but less likely to report breast reconstruction (P < 0.05). Satisfaction with care was correlated with patient-provider communication and overall HRQOL (P < 0.01) and physical, social/family, emotional and functional wellbeing (P < 0.01). SLP Latinas had lower emotional wellbeing than African-American YBCS (P < 0.01) and lower functional wellbeing than ELP Latina YBCS (P < 0.05). SLP Latina YBCS were less satisfied with their care compared with African-American and ELP Latina YBCS (P < 0.01). Financial toxicity seems to directly influence both access to care and quality care and survivorship outcomes. Investigating demographic characteristics and medical outcomes including HRQOL outcomes and satisfaction with care among ethnic minority YBCS is needed to advance the science as well as assist health professionals with precision care delivery. Greater translational and patient-centered research must focus on at-risk population such as YBCS to inform precision psychosocial oncology care and reduce health disparities. Copyright © 2018 John Wiley & Sons, Ltd.

Prediction of Carcass Composition Using Carcass Grading Traits in Hanwoo Steers.

PubMed

Lee, Jooyoung; Won, Seunggun; Lee, Jeongkoo; Kim, Jongbok

2016-09-01

The prediction of carcass composition in Hanwoo steers is very important for value-based marketing, and the improvement of prediction accuracy and precision can be achieved through the analyses of independent variables using a prediction equation with a sufficient dataset. The present study was conducted to develop a prediction equation for Hanwoo carcass composition for which data was collected from 7,907 Hanwoo steers raised at a private farm in Gangwon Province, South Korea, and slaughtered in the period between January 2009 and September 2014. Carcass traits such as carcass weight (CWT), back fat thickness (BFT), eye-muscle area (EMA), and marbling score (MAR) were used as independent variables for the development of a prediction equation for carcass composition, such as retail cut weight and percentage (RC, and %RC, respectively), trimmed fat weight and percentage (FAT, and %FAT, respectively), and separated bone weight and percentage (BONE, and %BONE), and its feasibility for practical use was evaluated using the estimated retail yield percentage (ELP) currently used in Korea. The equations were functions of all the variables, and the significance was estimated via stepwise regression analyses. Further, the model equations were verified by means of the residual standard deviation and the coefficient of determination (R(2)) between the predicted and observed values. As the results of stepwise analyses, CWT was the most important single variable in the equation for RC and FAT, and BFT was the most important variable for the equation of %RC and %FAT. The precision and accuracy of three variable equation consisting CWT, BFT, and EMA were very similar to those of four variable equation that included all for independent variables (CWT, BFT, EMA, and MAR) in RC and FAT, while the three variable equations provided a more accurate prediction for %RC. Consequently, the three-variable equation might be more appropriate for practical use than the four-variable equation based on its easy and cost-effective measurement. However, a relatively high average difference for the ELP in absolute value implies a revision of the official equation may be required, although the current official equation for predicting RC with three variables is still valid.

Polymeric Smart Skin Materials: Concepts, Materials, and Devices

DTIC Science & Technology

2006-03-31

nanotube actuators for both sensing and active control of surfaces. State-of-the-art OLED and photovoltaic materials have been developed for display...format. 14. SUBJECT TERMS Multi-sensor paints; carbon nanotube materials and devices; OLED , 15. NUMBER OF PAGES nhntovnlthir ndni elp.trAn-nntjc ’vicn...Significant advances in organic light emitting device ( OLED ) materials has also been achieved as is evident from the publications and invention

The "Common European Framework of Reference for Languages," the European Language Portfolio, and Language Teaching/Learning at University: An Argument and Some Proposals

ERIC Educational Resources Information Center

Little, David

2016-01-01

I begin this article by briefly explaining why I think CercleS should encourage university language centres to align their courses and assessment with the proficiency levels of the "Common European Framework of Reference for Languages" (CEFR) and why they should use a version of the European Language Portfolio (ELP) to support the…

The European Language Portfolio and Languages for Specific Purposes: A Project to Develop "Can Do" Descriptors Focused on Students' Interests and Motivation

ERIC Educational Resources Information Center

Gori, Federica

2014-01-01

Projects related to the European Language Portfolio (ELP) carried out at the Language Centre of the University of Trieste are concerned to examine whether and to what extent "can do" descriptors can be extended to courses in Languages for Specific Purposes. The project reported in this article had two aims. The first was to explore what…

Approaches to Automated Scoring of Speaking for K-12 English Language Proficiency Assessments. Policy Information Report and ETS Research Report Series No. RR-17-18

ERIC Educational Resources Information Center

Evanini, Keelan; Hauck, Maurice Cogan; Hakuta, Kenji

2017-01-01

This report is the fifth in a series concerning English language proficiency (ELP) assessments for English learners (ELs) in kindergarten through 12th grade in the United States. The series, produced by Educational Testing Service (ETS), is intended to provide theory and evidence-based principles and recommendations for improving next-generation…

Computational smart polymer design based on elastin protein mutability.

PubMed

Tarakanova, Anna; Huang, Wenwen; Weiss, Anthony S; Kaplan, David L; Buehler, Markus J

2017-05-01

Soluble elastin-like peptides (ELPs) can be engineered into a range of physical forms, from hydrogels and scaffolds to fibers and artificial tissues, finding numerous applications in medicine and engineering as "smart polymers". Elastin-like peptides are attractive candidates as a platform for novel biomaterial design because they exhibit a highly tunable response spectrum, with reversible phase transition capabilities. Here, we report the design of the first virtual library of elastin-like protein models using methods for enhanced sampling to study the effect of peptide chemistry, chain length, and salt concentration on the structural transitions of ELPs, exposing associated molecular mechanisms. We describe the behavior of the local molecular structure under increasing temperatures and the effect of peptide interactions with nearest hydration shell water molecules on peptide mobility and propensity to exhibit structural transitions. Shifts in the magnitude of structural transitions at the single-molecule scale are explained from the perspective of peptide-ion-water interactions in a library of four unique elastin-like peptide systems. Predictions of structural transitions are subsequently validated in experiment. This library is a valuable resource for recombinant protein design and synthesis as it elucidates mechanisms at the single-molecule level, paving a feedback path between simulation and experiment for smart material designs, with applications in biomedicine and diagnostic devices. Copyright © 2017. Published by Elsevier Ltd.

Bio-Orthogonally Crosslinked, Engineered Protein Hydrogels with Tunable Mechanics and Biochemistry for Cell Encapsulation.

PubMed

Madl, Christopher M; Katz, Lily M; Heilshorn, Sarah C

2016-06-07

Covalently-crosslinked hydrogels are commonly used as 3D matrices for cell culture and transplantation. However, the crosslinking chemistries used to prepare these gels generally cross-react with functional groups present on the cell surface, potentially leading to cytotoxicity and other undesired effects. Bio-orthogonal chemistries have been developed that do not react with biologically relevant functional groups, thereby preventing these undesirable side reactions. However, previously developed biomaterials using these chemistries still possess less than ideal properties for cell encapsulation, such as slow gelation kinetics and limited tuning of matrix mechanics and biochemistry. Here, engineered elastin-like proteins (ELPs) are developed that cross-link via strain-promoted azide-alkyne cycloaddition (SPAAC) or Staudinger ligation. The SPAAC-crosslinked materials form gels within seconds and complete gelation within minutes. These hydrogels support the encapsulation and phenotypic maintenance of human mesenchymal stem cells, human umbilical vein endothelial cells, and murine neural progenitor cells. SPAAC-ELP gels exhibit independent tuning of stiffness and cell adhesion, with significantly improved cell viability and spreading observed in materials containing a fibronectin-derived arginine-glycine-aspartic acid (RGD) domain. The crosslinking chemistry used permits further material functionalization, even in the presence of cells and serum. These hydrogels are anticipated to be useful in a wide range of applications, including therapeutic cell delivery and bioprinting.

Variability in gut mucosal secretory IgA in mice along a working day.

PubMed

Burns, Patricia; Oddi, Sofia; Forzani, Liliana; Tabacman, Eduardo; Reinheimer, Jorge; Vinderola, Gabriel

2018-02-05

To assess the variability of secretory immunoglobulin A (S-IgA) in the lumen and feces of mice along a working day. Mice were maintained under a 12 h light-dark cycle, light period starting at 8 AM. S-IgA was determined in feces and intestinal content (after one or three washes) at three points along the day: at the beginning, in the middle and at the end of the light period (ELP). Significant reduction in the content of S-IgA in the small intestine fluid and in feces was observed at the end of the light cycle, which coincides with the end of a regular working day (8 PM) in any given animal facility. It was also observed that three washes of the small intestine were more effective than one flush to recover a significant higher amount of S-IgA, with the smallest coefficient of variation observed by the ELP. A smaller CV would imply a reduced number of animals needed to achieve the same meaningful results. The results may be useful when designing animal trials for the selection of probiotic candidates based on their capacity of activating S-IgA, since it would imply a more rational use of experimental animals.

Intelligence: shared genetic basis between Mendelian disorders and a polygenic trait.

PubMed

Franić, Sanja; Groen-Blokhuis, Maria M; Dolan, Conor V; Kattenberg, Mathijs V; Pool, René; Xiao, Xiangjun; Scheet, Paul A; Ehli, Erik A; Davies, Gareth E; van der Sluis, Sophie; Abdellaoui, Abdel; Hansell, Narelle K; Martin, Nicholas G; Hudziak, James J; van Beijsterveldt, Catherina E M; Swagerman, Suzanne C; Hulshoff Pol, Hilleke E; de Geus, Eco J C; Bartels, Meike; Ropers, H Hilger; Hottenga, Jouke-Jan; Boomsma, Dorret I

2015-10-01

Multiple inquiries into the genetic etiology of human traits indicated an overlap between genes underlying monogenic disorders (eg, skeletal growth defects) and those affecting continuous variability of related quantitative traits (eg, height). Extending the idea of a shared genetic basis between a Mendelian disorder and a classic polygenic trait, we performed an association study to examine the effect of 43 genes implicated in autosomal recessive cognitive disorders on intelligence in an unselected Dutch population (N=1316). Using both single-nucleotide polymorphism (SNP)- and gene-based association testing, we detected an association between intelligence and the genes of interest, with genes ELP2, TMEM135, PRMT10, and RGS7 showing the strongest associations. This is a demonstration of the relevance of genes implicated in monogenic disorders of intelligence to normal-range intelligence, and a corroboration of the utility of employing knowledge on monogenic disorders in identifying the genetic variability underlying complex traits.

Cardiorespiratory, enzymatic and hormonal responses during and after walking while fasting

PubMed Central

Rosa, Claudio; Payan-Carreira, Rita; Lund, Rafael; Matos, Filipe; Garrido, Nuno

2018-01-01

The aim of the present study was to observe whether performing a low intensity endurance exercise following an overnight fasted (FAST) or fed (FED) condition promotes different cardiorespiratory, enzymatic and hormonal responses. Nine male physical active subjects, (age 21.89 ± 2.52 years old, height 175.89 ± 5.16 cm, weight 72.10 ± 4.31 kg, estimated body fat 7.25 ± 2.11%), randomly performed two sessions of 45 minutes’ low intensity exercise (individual ventilator threshold) interspersed by seven days, differentiated only in whether they were provided with a standardized meal or not. The oxygen consumption (VO2) and heart rate (HR) were measured continuously at the 30-min rest, the 45-min during and the 30-min post-exercise. The testosterone (T) and cortisol (C) hormones were measured at rest, immediately post-exercise and 15-min post-exercise. The Glucose (GLU), Free fatty acids (FFA) and enzyme lipase activity (ELP) were measured at rest, 15-min and 30-min exercise, immediately, 15-min and 30-min post-exercise. Significantly lower values were observed in FED compared to FAST with: C (nmol/L) from pre (428.87 ± 120.41; 454.62 ± 148.33, respectively) to immediately post-exercise (285.10 ± 85.86; 465.66 ± 137.70, respectively) and 15-min post-exercise (248.00 ± 87.88; 454.31 ± 112.72, respectively) (p<0.05); and GLU at all times, with an exception at 15-min post-exercise. The testosterone/cortisol ratio (T/C) was significantly higher in the FED compared with FAST from pre (0.05 ± 0.02, 0.05 ± 0.01, respectively) to 15-min post-exercise (0.08 ± 0.03, 0.05 ± 0.02, respectively). No other significant differences were observed between conditions. We conclude that fasting prior to low intensity endurance exercise does not seem be advantageous, when it comes to fat loss, compared with the same exercise performed after a meal. PMID:29494664

National Health Insurance by Regulation: Mandated Employee Benefits,

DTIC Science & Technology

1980-04-01

A0AO95 050 RANW CORP SANTA MONICA CA F/0 S/I1 NATIONAL HEALTH INSURANCE BY REKULATION: MANDATED EMPLOYEE NE-TC(U) APR 80 C E PI4ELPS LICLASSIFIED...31 ! 9 : I NATIONAL HEALTH INSURANCE BY REGULATION: MANDATED EMPLOYEE BENEFITS 1 I. INTRODUCTION Social issues have often been solved...offer a variety of insurance packages to employees , iThis paper was presented at the Conference on "National Health Insurance: Ihat Now, What Later, What

In planta production of ELPylated spidroin-based proteins results in non-cytotoxic biopolymers.

PubMed

Hauptmann, Valeska; Menzel, Matthias; Weichert, Nicola; Reimers, Kerstin; Spohn, Uwe; Conrad, Udo

2015-02-19

Spider silk is a tear-resistant and elastic biopolymer that has outstanding mechanical properties. Additionally, exiguous immunogenicity is anticipated for spider silks. Therefore, spider silk represents a potential ideal biomaterial for medical applications. All known spider silk proteins, so-called spidroins, reveal a composite nature of silk-specific units, allowing the recombinant production of individual and combined segments. In this report, a miniaturized spidroin gene, named VSO1 that contains repetitive motifs of MaSp1 has been synthesized and combined to form multimers of distinct lengths, which were heterologously expressed as elastin-like peptide (ELP) fusion proteins in tobacco. The elastic penetration moduli of layered proteins were analyzed for different spidroin-based biopolymers. Moreover, we present the first immunological analysis of synthetic spidroin-based biopolymers. Characterization of the binding behavior of the sera after immunization by competitive ELISA suggested that the humoral immune response is mainly directed against the fusion partner ELP. In addition, cytocompatibility studies with murine embryonic fibroblasts indicated that recombinant spidroin-based biopolymers, in solution or as coated proteins, are well tolerated. The results show that spidroin-based biopolymers can induce humoral immune responses that are dependent on the fusion partner and the overall protein structure. Furthermore, cytocompatibility assays gave no indication of spidroin-derived cytotoxicity, suggesting that recombinant produced biopolymers composed of spider silk-like repetitive elements are suitable for biomedical applications.

Light Scattering Characterization of Elastin-Like Polypeptide Trimer Micelles

NASA Astrophysics Data System (ADS)

Tsuper, Ilona; Terrano, Daniel; Maraschky, Adam; Holland, Nolan; Streletzky, Kiril

The elastin-like polypeptides (ELP) nanoparticles are composed of three-armed star polypeptides connected by a negatively charged foldon. Each of the three arms extending from the foldon domain includes 20 repeats of the (GVGVP) amino acid sequence. The ELP polymer chains are soluble at room temperature and become insoluble at the transition temperature (close to 50 ° C), forming micelles. The size and shape of the micelle are dependent on the temperature and the pH of the solution, and on the concentration of the phosphate buffered saline (PBS). The depolarized dynamic light scattering (DDLS) was employed to study the structure and dynamics of micelles at 62 ° C. The solution was maintained at an approximate pH level of 7.3 - 7.5, while varying PBS concentration. At low salt concentrations (<15 mM), the micelle radius was about 10nm but not very reproducible on account of unstable pH levels arising from low buffer concentrations. At intermediate salt concentrations (15 - 60 mM), the system formed spherically-shaped micelles, exhibiting a steady growth in the hydrodynamic radius (Rh) from 10 to 21 nm, with increasing PBS concentration. Interestingly, higher salt concentrations (>60 mM) displayed an apparent elongation of the micelles evident by a significant VH signal, along with a surge in the apparent Rh. A model of micelle growth (and potential elongation) with increase in salt concentration is considered.

Sequence Directionality Dramatically Affects LCST Behavior of Elastin-Like Polypeptides.

PubMed

Li, Nan K; Roberts, Stefan; Quiroz, Felipe Garcia; Chilkoti, Ashutosh; Yingling, Yaroslava G

2018-04-30

Elastin-like polypeptides (ELP) exhibit an inverse temperature transition or lower critical solution temperature (LCST) transition phase behavior in aqueous solutions. In this paper, the thermal responsive properties of the canonical ELP, poly(VPGVG), and its reverse sequence poly(VGPVG) were investigated by turbidity measurements of the cloud point behavior, circular dichroism (CD) measurements, and all-atom molecular dynamics (MD) simulations to gain a molecular understanding of mechanism that controls hysteretic phase behavior. It was shown experimentally that both poly(VPGVG) and poly(VGPVG) undergo a transition from soluble to insoluble in aqueous solution upon heating above the transition temperature ( T t ). However, poly(VPGVG) resolubilizes upon cooling below its T t , whereas the reverse sequence, poly(VGPVG), remains aggregated despite significant undercooling below the T t . The results from MD simulations indicated that a change in sequence order results in significant differences in the dynamics of the specific residues, especially valines, which lead to extensive changes in the conformations of VPGVG and VGPVG pentamers and, consequently, dissimilar propensities for secondary structure formation and overall structure of polypeptides. These changes affected the relative hydrophilicities of polypeptides above T t , where poly(VGPVG) is more hydrophilic than poly(VPGVG) with more extended conformation and larger surface area, which led to formation of strong interchain hydrogen bonds responsible for stabilization of the aggregated phase and the observed thermal hysteresis for poly(VGPVG).

Formal expressions and corresponding expansions for the exact Kohn-Sham exchange potential

NASA Astrophysics Data System (ADS)

Bulat, Felipe A.; Levy, Mel

2009-11-01

Formal expressions and their corresponding expansions in terms of Kohn-Sham (KS) orbitals are deduced for the exchange potential vx(r) . After an alternative derivation of the basic optimized effective potential integrodifferential equations is given through a Hartree-Fock adiabatic connection perturbation theory, we present an exact infinite expansion for vx(r) that is particularly simple in structure. It contains the very same occupied-virtual quantities that appear in the well-known optimized effective potential integral equation, but in this new expression vx(r) is isolated on one side of the equation. An orbital-energy modified Slater potential is its leading term which gives encouraging numerical results. Along different lines, while the earlier Krieger-Li-Iafrate approximation truncates completely the necessary first-order perturbation orbitals, we observe that the improved localized Hartree-Fock (LHF) potential, or common energy denominator potential (CEDA), or effective local potential (ELP), incorporates the part of each first-order orbital that consists of the occupied KS orbitals. With this in mind, the exact correction to the LHF, CEDA, or ELP potential (they are all equivalent) is deduced and displayed in terms of the virtual portions of the first-order orbitals. We close by observing that the newly derived exact formal expressions and corresponding expansions apply as well for obtaining the correlation potential from an orbital-dependent correlation energy functional.

Ex-Vivo Lymphatic Perfusion System for Independently Controlling Pressure Gradient and Transmural Pressure in Isolated Vessels

PubMed Central

Kornuta, Jeffrey A.; Dixon, J. Brandon

2015-01-01

In addition to external forces, collecting lymphatic vessels intrinsically contract to transport lymph from the extremities to the venous circulation. As a result, the lymphatic endothelium is routinely exposed to a wide range of dynamic mechanical forces, primarily fluid shear stress and circumferential stress, which have both been shown to affect lymphatic pumping activity. Although various ex-vivo perfusion systems exist to study this innate pumping activity in response to mechanical stimuli, none are capable of independently controlling the two primary mechanical forces affecting lymphatic contractility: transaxial pressure gradient, ΔP, which governs fluid shear stress; and average transmural pressure, Pavg, which governs circumferential stress. Hence, the authors describe a novel ex-vivo lymphatic perfusion system (ELPS) capable of independently controlling these two outputs using a linear, explicit model predictive control (MPC) algorithm. The ELPS is capable of reproducing arbitrary waveforms within the frequency range observed in the lymphatics in vivo, including a time-varying ΔP with a constant Pavg, time-varying ΔP and Pavg, and a constant ΔP with a time-varying Pavg. In addition, due to its implementation of syringes to actuate the working fluid, a post-hoc method of estimating both the flow rate through the vessel and fluid wall shear stress over multiple, long (5 sec) time windows is also described. PMID:24809724

Recent hybridization between Taenia asiatica and Taenia saginata.

PubMed

Yamane, Kanako; Suzuki, Yumi; Tachi, Eiko; Li, Tiaoying; Chen, Xingwang; Nakao, Minoru; Nkouawa, Agathe; Yanagida, Testuya; Sako, Yasuhito; Ito, Akira; Sato, Hiroshi; Okamoto, Munehiro

2012-06-01

Five Taenia tapeworms collected from humans in Tibetan Plateau, Sichuan, China, where three species of human Taenia are sympatrically endemic, were examined for the mitochondrial cox1 gene and two nuclear genes, ef1 and elp. Phylogenetic analyses of these genes revealed that two adult worms showed nuclear-mitochondrial discordance, suggesting that they originated from hybridization between Taenia saginata and Taenia asiatica. One of two worms had T. asiatica-type mtDNA, whereas another worm had T. saginata-type mtDNA, indicating that reciprocal hybridization between T. saginata and T. asiatica could occur. The worm having T. asiatica-type mtDNA was heterozygous at both nuclear loci with T. saginata-type alleles and T. asiatica-type alleles. In another worm, the ef1 locus was heterozygous with a T. saginata-type alleles and T. asiatica-type alleles, while the elp locus was homozygous with T. saginata-type alleles. Self-fertilization is the main reproductive method of the genus Taenia. Since self-fertilization represents a type of inbreeding, each locus in the offspring would become homozygous over generations with genetic drift. The fact that some nuclear loci are still heterozygous means that hybridization might have occurred recently. Hybridization between T. asiatica and T. saginata is probably an ongoing event in many areas in which they are sympatrically endemic. Crown Copyright © 2012. Published by Elsevier Ireland Ltd. All rights reserved.

Designing protein-based biomaterials for medical applications.

PubMed

Gagner, Jennifer E; Kim, Wookhyun; Chaikof, Elliot L

2014-04-01

Biomaterials produced by nature have been honed through billions of years, evolving exquisitely precise structure-function relationships that scientists strive to emulate. Advances in genetic engineering have facilitated extensive investigations to determine how changes in even a single peptide within a protein sequence can produce biomaterials with unique thermal, mechanical and biological properties. Elastin, a naturally occurring protein polymer, serves as a model protein to determine the relationship between specific structural elements and desirable material characteristics. The modular, repetitive nature of the protein facilitates the formation of well-defined secondary structures with the ability to self-assemble into complex three-dimensional architectures on a variety of length scales. Furthermore, many opportunities exist to incorporate other protein-based motifs and inorganic materials into recombinant protein-based materials, extending the range and usefulness of these materials in potential biomedical applications. Elastin-like polypeptides (ELPs) can be assembled into 3-D architectures with precise control over payload encapsulation, mechanical and thermal properties, as well as unique functionalization opportunities through both genetic and enzymatic means. An overview of current protein-based materials, their properties and uses in biomedicine will be provided, with a focus on the advantages of ELPs. Applications of these biomaterials as imaging and therapeutic delivery agents will be discussed. Finally, broader implications and future directions of these materials as diagnostic and therapeutic systems will be explored. Copyright © 2013 Elsevier Ltd. All rights reserved.

Elongator-dependent modification of cytoplasmic tRNALysUUU is required for mitochondrial function under stress conditions

PubMed Central

Tigano, Marco; Ruotolo, Roberta; Dallabona, Cristina; Fontanesi, Flavia; Barrientos, Antoni; Donnini, Claudia; Ottonello, Simone

2015-01-01

To gain a wider view of the pathways that regulate mitochondrial function, we combined the effect of heat stress on respiratory capacity with the discovery potential of a genome-wide screen in Saccharomyces cerevisiae. We identified 105 new genes whose deletion impairs respiratory growth at 37°C by interfering with processes such as transcriptional regulation, ubiquitination and cytosolic tRNA wobble uridine modification via 5-methoxycarbonylmethyl-2-thiouridine formation. The latter process, specifically required for efficient decoding of AA-ending codons under stress conditions, was covered by multiple genes belonging to the Elongator (e.g. ELP3) and urmylation (e.g., NCS6) pathways. ELP3 or NCS6 deletants had impaired mitochondrial protein synthesis. Their respiratory deficiency was selectively rescued by overexpression of tRNALysUUU as well by overexpression of genes (BCK1 and HFM1) with a strong bias for the AAA codon read by this tRNA. These data extend the mitochondrial regulome, demonstrate that heat stress can impair respiration by disturbing cytoplasmic translation of proteins critically involved in mitochondrial function and document, for the first time, the involvement in such process of the Elongator and urmylation pathways. Given the conservation of these pathways, the present findings may pave the way to a better understanding of the human mitochondrial regulome in health and disease. PMID:26240381

Test Takers' Writing Activities during the "TOEFL iBT"® Writing Tasks: A Stimulated Recall Study. "TOEFL iBT"® Research Report. TOEFL iBT-25. ETS Research Report No. RR-15-04

ERIC Educational Resources Information Center

Barkaoui, Khaled

2015-01-01

This study aimed to describe the writing activities that test takers engage in when responding to the writing tasks in the "TOEFL iBT"[superscript R] test and to examine the effects of task type and test-taker English language proficiency (ELP) and keyboarding skills on the frequency and distribution of these activities. Each of 22 test…

A Correlational Study of Graphic Organizers and Science Achievement of English Language Learners

NASA Astrophysics Data System (ADS)

Clarke, William Gordon

English language learners (ELLs) demonstrate lower academic performance and have lower graduation and higher dropout rates than their non-ELL peers. The primary purpose of this correlational quantitative study was to investigate the relationship between the use of graphic organizer-infused science instruction and science learning of high school ELLs. Another objective was to determine if the method of instruction, socioeconomic status (SES), gender, and English language proficiency (ELP) were predictors of academic achievement of high school ELLs. Data were gathered from a New York City (NYC) high school fall 2012-2013 archival records of 145 ninth-grade ELLs who had received biology instruction in freestanding English as a second language (ESL) classes, followed by a test of their learning of the material. Fifty-four (37.2%) of these records were of students who had learned science by the conventional textbook method, and 91 (62.8%) by using graphic organizers. Data analysis employed the Statistical Package for the Social Sciences (SPSS) software for multiple regression analysis, which found graphic organizer use to be a significant predictor of New York State Regents Living Environment (NYSRLE) test scores (p < .01). One significant regression model was returned whereby, when combined, the four predictor variables (method of instruction, SES, gender, and ELP) explained 36% of the variance of the NYSRLE score. Implications of the study findings noted graphic organizer use as advantageous for ELL science achievement. Recommendations made for practice were for (a) the adoption of graphic organizer infused-instruction, (b) establishment of a protocol for the implementation of graphic organizer-infused instruction, and (c) increased length of graphic organizer instructional time. Recommendations made for future research were (a) a replication quantitative correlational study in two or more high schools, (b) a quantitative quasi-experimental quantitative study to determine the influence of graphic organizer instructional intervention and ELL science achievement, (c) a quantitative quasi-experimental study to determine the effect of teacher-based factors on graphic organizer-infused instruction, and (c) a causal comparative study to determine the efficacy of graphic organizer use in testing modifications for high school ELL science.

The Clinician as Leader: Why, How, and When.

PubMed

Stoller, James K

2017-11-01

Clinicians are inveterate leaders. We lead patients through the difficult maze of illness, families through the travails of ill loved ones, and physicians-in-training through the gauntlet of learning medicine. Yet, in the context of a range of leadership styles that effective leaders must be able to deploy situationally, physician leaders have traditionally defaulted to a "command and control" style that fosters the concept of physicians as "Viking warriors" or "heroic lone healers." The perverse effects of "command and control" are that this style conspires against collaboration and tends to be perpetuated as aspiring leaders emulate their predecessors. Because healthcare faces challenges of cost, access, and quality and is in the throes of change, the current landscape requires effective leadership. Though still relatively uncommon among healthcare organizations, frontrunner organizations are offering leadership development programs. The design of such programs requires clarity about requisite leadership competencies and about how and when to best to deliver such curricula. As one example, the American Thoracic Society has launched its Emerging Leaders Program (ELP), which is currently offering a leadership development curriculum to 18 selected emerging leaders. The ATS ELP curriculum focuses on awareness of self and system and incorporates highly participatory sessions on emotional intelligence, teambuilding, change management, situational leadership, appreciative inquiry, process and quality improvement, strategic planning, and organizational culture. Short-term deliverables are the development and presentation of business plans for innovations proposed by the group. Hoped for longer-term outcomes include an enhanced leadership pipeline for global respiratory health.

Elongator-dependent modification of cytoplasmic tRNALysUUU is required for mitochondrial function under stress conditions.

PubMed

Tigano, Marco; Ruotolo, Roberta; Dallabona, Cristina; Fontanesi, Flavia; Barrientos, Antoni; Donnini, Claudia; Ottonello, Simone

2015-09-30

To gain a wider view of the pathways that regulate mitochondrial function, we combined the effect of heat stress on respiratory capacity with the discovery potential of a genome-wide screen in Saccharomyces cerevisiae. We identified 105 new genes whose deletion impairs respiratory growth at 37°C by interfering with processes such as transcriptional regulation, ubiquitination and cytosolic tRNA wobble uridine modification via 5-methoxycarbonylmethyl-2-thiouridine formation. The latter process, specifically required for efficient decoding of AA-ending codons under stress conditions, was covered by multiple genes belonging to the Elongator (e.g. ELP3) and urmylation (e.g., NCS6) pathways. ELP3 or NCS6 deletants had impaired mitochondrial protein synthesis. Their respiratory deficiency was selectively rescued by overexpression of tRNA(Lys) UUU as well by overexpression of genes (BCK1 and HFM1) with a strong bias for the AAA codon read by this tRNA. These data extend the mitochondrial regulome, demonstrate that heat stress can impair respiration by disturbing cytoplasmic translation of proteins critically involved in mitochondrial function and document, for the first time, the involvement in such process of the Elongator and urmylation pathways. Given the conservation of these pathways, the present findings may pave the way to a better understanding of the human mitochondrial regulome in health and disease. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

Cellular biophysics during freezing of rat and mouse sperm predicts post-thaw motility.

PubMed

Hagiwara, Mie; Choi, Jeung Hwan; Devireddy, Ramachandra V; Roberts, Kenneth P; Wolkers, Willem F; Makhlouf, Antoine; Bischof, John C

2009-10-01

Though cryopreservation of mouse sperm yields good survival and motility after thawing, cryopreservation of rat sperm remains a challenge. This study was designed to evaluate the biophysics (membrane permeability) of rat in comparison to mouse to better understand the cooling rate response that contributes to cryopreservation success or failure in these two sperm types. In order to extract subzero membrane hydraulic permeability in the presence of ice, a differential scanning calorimeter (DSC) method was used. By analyzing rat and mouse sperm frozen at 5 degrees C/min and 20 degrees C/min, heat release signatures characteristic of each sperm type were obtained and correlated to cellular dehydration. The dehydration response was then fit to a model of cellular water transport (dehydration) by adjusting cell-specific biophysical (membrane hydraulic permeability) parameters L(pg) and E(Lp). A "combined fit" (to 5 degrees C/min and 20 degrees C/min data) for rat sperm in Biggers-Whitten-Whittingham media yielded L(pg) = 0.007 microm min(-1) atm(-1) and E(Lp) = 17.8 kcal/mol, and in egg yolk cryopreservation media yielded L(pg) = 0.005 microm min(-1) atm(-1) and E(Lp) = 14.3 kcal/mol. These parameters, especially the activation energy, were found to be lower than previously published parameters for mouse sperm. In addition, the biophysical responses in mouse and rat sperm were shown to depend on the constituents of the cryopreservation media, in particular egg yolk and glycerol. Using these parameters, optimal cooling rates for cryopreservation were predicted for each sperm based on a criteria of 5%-15% normalized cell water at -30 degrees C during freezing in cryopreservation media. These predicted rates range from 53 degrees C/min to 70 degrees C/min and from 28 degrees C/min to 36 degrees C/min in rat and mouse, respectively. These predictions were validated by comparison to experimentally determined cryopreservation outcomes, in this case based on motility. Maximum motility was obtained with freezing rates between 50 degrees C/min and 80 degrees C/min for rat and at 20 degrees C/min with a sharp drop at 50 degrees C/min for mouse. In summary, DSC experiments on mouse and rat sperm yielded a difference in membrane permeability parameters in the two sperm types that, when implemented in a biophysical model of water transport, reasonably predict different optimal cooling rate outcomes for each sperm after cryopreservation.

The Polychromatic Laser Guide Star: the ELP-OA demonstrator at Observatoire de Haute Provence

NASA Astrophysics Data System (ADS)

Foy, R.; Chatagnat, M.; Dubet, D.; Éric, P.; Eysseric, J.; Foy, F.-C.; Fusco, T.; Girard, J.; Laloge, A.; Le van Suu, A.; Messaoudi, B.; Perruchot, S.; Richaud, P.; Richaud, Y.; Rondeau, X.; Tallon, M.; Thiébaut, É.; Boër, M.

2007-07-01

The correction of the tilt for adaptive optics devices from the only laser guide star can be done with the polychromatic laser guide star. We report the progress of the first demonstrator of the implementation of this concept, at Observatoire de Haute-Provence. We review the last steps of the feasibility studies, the optimization of the laser parameters, and the studies of the implementation at the OHP 1.52m telescope, including the beam propagation to the lasers room to the mesosphere and the algorithms for tip-tilt measurements.

A 2017 stellar occultation by Orcus/Vanth

NASA Astrophysics Data System (ADS)

Sickafoose, Amanda A.; Bosh, Amanda S.; Levine, Stephen; Zuluaga, Carlos A.; Genade, Anja; Schindler, Karsten; Lister, Tim; Person, Michael J.

2017-10-01

(90482) Orcus is a large trans-Neptunian object (TNO) of diameter ~900 km, located in the 3:2 orbital resonance with Neptune. This plutino has a satellite, Vanth, approximately 280 km in diameter. Vanth orbits roughly 9000 km from Orcus in a ~9.5-day period. This system is particularly interesting, as Orcus falls between the small, spectrally-bland TNOs and the large TNOs having spectra rich in volatile features, while Vanth might have resulted from either collision or capture.A stellar occultation by Orcus was predicted to occur on 07 March 2017. Observations were made from five sites: the 0.6-m Astronomical Telescope of the University of Stuttgart (ATUS) at Sierra Remote Observatories (SRO), California; Las Cumbres Observatory’s 1-m telescope (ELP) at McDonald Observatory, Fort Davis, Texas; NASA’s 3-m InfraRed Telescope Facility (IRTF) on Mauna Kea, Hawaii; the 4.1-m Southern Astrophysical Research telescope (SOAR) on Cerro Pachón, Chile; and the 0.6-m Southeastern Association for Research in Astronomy telescope (SARA-CT) at Cerro Tololo, Chile. High-speed, visible-wavelength images were taken at all sites, in addition to simultaneous K-band images at the IRTF. A solid-body occultation was observed at both ELP and the IRTF. Offset midtimes and incompatible light ratios suggest that two different stars were occulted by two different bodies, likely Orcus and Vanth. See Bosh et al. this conference for details of the astrometry for the event. Here, we present results from the observations, including light curves, size and albedo estimates, and upper limits on a possible atmosphere.

The United States Air Force in Southeast Asia. The War in South Vietnam. The Years of the Offensive 1965-1968

DTIC Science & Technology

1988-01-01

principle of introducing an international force, he had to discuss it with Maj. Gen. Tran Van Minh and Maj. Gen. Nguyen Van Thieu of the Armed Forces...from it within 2 days. Another attack in August on the camp at Duc Co was beaten back, largely by 280 tactical strike sorties. The final major enemy... Duc Co CORPS Nha Trang [3 CORPS Cam Ranh Bay Loc Ninh - 0 operation )’ Elp 4’Wng Vl pTo _fongN Ar0 ofPrloqe Cnfonat AttleMlbaroorroourdr ar Bate 138

Initial clinical experience with a radiation oncology dedicated open 1.0T MR‐simulation

PubMed Central

Wen, Ning; Hearshen, David; Kim, Joshua; Pantelic, Milan; Zhao, Bo; Mancell, Tina; Levin, Kenneth; Movsas, Benjamin; Chetty, Indrin J.; Siddiqui, M. Salim

2015-01-01

The purpose of this study was to describe our experience with 1.0T MR‐SIM including characterization, quality assurance (QA) program, and features necessary for treatment planning. Staffing, safety, and patient screening procedures were developed. Utilization of an external laser positioning system (ELPS) and MR‐compatible couchtop were illustrated. Spatial and volumetric analyses were conducted between CT‐SIM and MR‐SIM using a stereotactic QA phantom with known landmarks and volumes. Magnetic field inhomogeneity was determined using phase difference analysis. System‐related, in‐plane distortion was evaluated and temporal changes were assessed. 3D distortion was characterized for regions of interest (ROIs) 5–20 cm away from isocenter. American College of Radiology (ACR) recommended tests and impact of ELPS on image quality were analyzed. Combined ultrashort echotime Dixon (UTE/Dixon) sequence was evaluated. Amplitude‐triggered 4D MRI was implemented using a motion phantom (2–10 phases, ~2 cm excursion, 3–5 s periods) and a liver cancer patient. Duty cycle, acquisition time, and excursion were evaluated between maximum intensity projection (MIP) datasets. Less than 2% difference from expected was obtained between CT‐SIM and MR‐SIM volumes, with a mean distance of <0.2 mm between landmarks. Magnetic field inhomogeneity was <2 ppm. 2D distortion was <2 mm over 28.6–33.6 mm of isocenter. Within 5 cm radius of isocenter, mean 3D geometric distortion was 0.59±0.32 mm (maximum=1.65 mm) and increased 10–15 cm from isocenter (mean=1.57±1.06 mm, maximum=6.26 mm). ELPS interference was within the operating frequency of the scanner and was characterized by line patterns and a reduction in signal‐to‐noise ratio (4.6–12.6% for TE=50−150 ms). Image quality checks were within ACR recommendations. UTE/Dixon sequences yielded detectability between bone and air. For 4D MRI, faster breathing periods had higher duty cycles than slow (50.4% (3 s) and 39.4% (5 s), p<0.001) and ~ fourfold acquisition time increase was measured for ten‐phase versus two‐phase. Superior–inferior object extent was underestimated 8% (6 mm) for two‐phase as compared to ten‐phase MIPs, although <2% difference was obtained for ≥4 phases. 4D MRI for a patient demonstrated acceptable image quality in ~7 min. MR‐SIM was integrated into our workflow and QA procedures were developed. Clinical applicability was demonstrated for 4D MRI and UTE imaging to support MR‐SIM for single modality treatment planning. PACS numbers: 87.56.Fc, 87.61.‐c, 87.57.cp PMID:26103190

Production of the Main Celiac Disease Autoantigen by Transient Expression in Nicotiana benthamiana

PubMed Central

Marín Viegas, Vanesa S.; Acevedo, Gonzalo R.; Bayardo, Mariela P.; Chirdo, Fernando G.; Petruccelli, Silvana

2015-01-01

Celiac Disease (CD) is a gluten sensitive enteropathy that remains widely undiagnosed and implementation of massive screening tests is needed to reduce the long term complications associated to untreated CD. The main CD autoantigen, human tissue transglutaminase (TG2), is a challenge for the different expression systems available since its cross-linking activity affects cellular processes. Plant-based transient expression systems can be an alternative for the production of this protein. In this work, a transient expression system for the production of human TG2 in Nicotiana benthamiana leaves was optimized and reactivity of plant-produced TG2 in CD screening test was evaluated. First, a subcellular targeting strategy was tested. Cytosolic, secretory, endoplasmic reticulum (C-terminal SEKDEL fusion) and vacuolar (C-terminal KISIA fusion) TG2 versions were transiently expressed in leaves and recombinant protein yields were measured. ER-TG2 and vac-TG2 levels were 9- to 16-fold higher than their cytosolic and secretory counterparts. As second strategy, TG2 variants were co-expressed with a hydrophobic elastin-like polymer (ELP) construct encoding for 36 repeats of the pentapeptide VPGXG in which the guest residue X were V and F in ratio 8:1. Protein bodies (PB) were induced by the ELP, with a consequent two-fold-increase in accumulation of both ER-TG2 and vac-TG2. Subsequently, ER-TG2 and vac-TG2 were produced and purified using immobilized metal ion affinity chromatography. Plant purified ER-TG2 and vac-TG2 were recognized by three anti-TG2 monoclonal antibodies that bind different epitopes proving that plant-produced antigen has immunochemical characteristics similar to those of human TG2. Lastly, an ELISA was performed with sera of CD patients and healthy controls. Both vac-TG2 and ER-TG2 were positively recognized by IgA of CD patients while they were not recognized by serum from non-celiac controls. These results confirmed the usefulness of plant-produced TG2 to develop screening assays. In conclusion, the combination of subcellular sorting strategy with co-expression with a PB inducing construct was sufficient to increase TG2 protein yields. This type of approach could be extended to other problematic proteins, highlighting the advantages of plant based production platforms. PMID:26648956

A conserved modified wobble nucleoside (mcm5s2U) in lysyl-tRNA is required for viability in yeast

PubMed Central

Björk, Glenn R.; Huang, Bo; Persson, Olof P.; Byström, Anders S.

2007-01-01

Transfer RNAs specific for Gln, Lys, and Glu from all organisms (except Mycoplasma) and organelles have a 2-thiouridine derivative (xm5s2U) as wobble nucleoside. These tRNAs read the A- and G-ending codons in the split codon boxes His/Gln, Asn/Lys, and Asp/Glu. In eukaryotic cytoplasmic tRNAs the conserved constituent (xm5-) in position 5 of uridine is 5-methoxycarbonylmethyl (mcm5). A protein (Tuc1p) from yeast resembling the bacterial protein TtcA, which is required for the synthesis of 2-thiocytidine in position 32 of the tRNA, was shown instead to be required for the synthesis of 2-thiouridine in the wobble position (position 34). Apparently, an ancient member of the TtcA family has evolved to thiolate U34 in tRNAs of organisms from the domains Eukarya and Archaea. Deletion of the TUC1 gene together with a deletion of the ELP3 gene, which results in the lack of the mcm5 side chain, removes all modifications from the wobble uridine derivatives of the cytoplasmic tRNAs specific for Gln, Lys, and Glu, and is lethal to the cell. Since excess of the unmodified form of these three tRNAs rescued the double mutant elp3 tuc1, the primary function of mcm5s2U34 seems to be to improve the efficiency to read the cognate codons rather than to prevent mis-sense errors. Surprisingly, overexpression of the mcm5s2U-lacking tRNALys alone was sufficient to restore viability of the double mutant. PMID:17592039

Climate of Earth-Like Planets With and Without Ocean Heat Transport Orbiting a Range of M and K Stars

NASA Technical Reports Server (NTRS)

Kiang, N. Y.; Jablonski, Emma R.; Way, Michael J.; Del Genio, Anthony; Roberge, Aki

2015-01-01

The mean surface temperature of a planet is now acknowledged as insufficient to surmise its full potential habitability. Advancing our understanding requires exploration with 3D general circulation models (GCMs), which can take into account how gradients and fluxes across a planet's surface influence the distribution of heat, clouds, and the potential for heterogeneous distribution of liquid water. Here we present 3D GCM simulations of the effects of alternative stellar spectra, instellation, model resolution, and ocean heat transport, on the simulated distribution of heat and moisture of an Earth-like planet (ELP).

Structural Basis for Flip-Flop Action of Thiamin Pyrophosphate-Dependent Enzymes Revealed by Human Pyruvate Dehydrogenase

NASA Technical Reports Server (NTRS)

Dominiak, Paulina; Ciszak, Ewa M.; Korotchkina, Lioubov; Sidhu, Sukhdeep; Patel, Mulchand

2003-01-01

Thiamin pyrophosphate (TPP), the biologically active form of vitamin BI, is a cofactor of enzymes catalyzing reactions involving the cleavage of a carbon-carbon bond adjacent to an oxo group. TPP-dependent enzymes show a common mechanism of TPP activation by: (1) forming the ionic N-H...O(sup -) hydrogen bonding between the N1' atom of the aminopirymidine ring of the coenzyme and intrinsic gamma-carboxylate group of glutamate and (2) imposing an "active" V-conformation that brings the N4' atom of the aminopirymidine to the distance required for the intramolecular C-H.. .N hydrogen bonding with the thiazolium C2 atom. Within these two hydrogen bonds that rapidly exchange protons, protonation of the N1' atom is strictly coordinated with the deprotonation of the 4' -amino group and eventually abstraction of the proton from C2. The human pyruvate dehydrogenase Elp, component of human pyruvate dehydrogenase complex, catalyzes the irreversible decarboxylation of the pyruvate followed by the reductive acetylation of the lipoyl group of dihydrolipoyl acyltransferase. Elp is alpha(sub 2)beta(sub2)-heterotetrameric with a molecular mass of I54 kDa, which has two catalytic sites, each providing TPP and magnesium ion as cofactors and each formed on the interface between the PP and PYR domains. The dynamic nonequivalence of two otherwise chemically equivalent catalytic sites has been observed and the flip-flop mechanism was suggested, according to which two active sites affect each other and in which different steps of the catalytic reaction are performed in each of the sites at any given moment. Based on specific futures of human pyruvate dehydrogenase including rigid and flexible connections between domains that bind the cofactor we propose a mechanistic model for the flip-flop action of this enzyme. We postulate that the dynamic protein environment drives the exchange of tautomers in the 4' -aminopyrimidine ring of the cofactor through a concerted shuttl-like motion of tightly connected domains. The dynamic exchange of those tautomers, in turns, is required during the reactions of pyruvate decarboxylation and reductive acetylation of lipoamide. Thus the shuttle-like motion of the domains is coordinated with the reactions of decarboxylation and acetylation, which are carried out in each of the cofactor sites resulting in a flip-flop action of the enzyme. The structure-derived mechanism of action of human pyruvate dehydrogenase may be likely common for other TPP-dependent enzymes.

Intra-vitreal αB crystallin fused to elastin-like polypeptide provides neuroprotection in a mouse model of age-related macular degeneration.

PubMed

Sreekumar, Parameswaran G; Li, Zhe; Wang, Wan; Spee, Christine; Hinton, David R; Kannan, Ram; MacKay, J Andrew

2018-05-18

Age-related macular degeneration (AMD) is the leading cause of severe and irreversible central vision loss, and the primary site of AMD pathology is the retinal pigment epithelium (RPE). Geographic atrophy (GA) is an advanced form of AMD characterized by extensive RPE cell loss, subsequent degeneration of photoreceptors, and thinning of retina. This report describes the protective potential of a peptide derived from the αB crystallin protein using a sodium iodate (NaIO 3 ) induced mouse model of GA. Systemic NaIO 3 challenge causes degeneration of the RPE and neighboring photoreceptors, which have similarities to retinas of GA patients. αB crystallin is an abundant ocular protein that maintains ocular clarity and retinal homeostasis, and a small peptide from this protein (mini cry) displays neuroprotective properties. To retain this peptide for longer in the vitreous, mini cry was fused to an elastin-like polypeptide (ELP). A single intra-vitreal treatment by this crySI fusion significantly inhibits retinal degeneration in comparison to free mini cry. While mini cry is cleared from the eye with a mean residence time of 0.4 days, crySI is retained with a mean residence time of 3.0 days; furthermore, fundus photography reveals evidence of retention at two weeks. Unlike the free mini cry, crySI protects the RPE against NaIO 3 challenge for at least two weeks after administration. CrySI also inhibits RPE apoptosis and caspase-3 activation and protects the retina from cell death up to 1-month post NaIO 3 challenge. These results show that intra-ocular ELP-linked peptides such as crySI hold promise as protective agents to prevent RPE atrophy and progressive retinal degeneration in AMD. Copyright © 2018 Elsevier B.V. All rights reserved.

Elongator complex influences telomeric gene silencing and DNA damage response by its role in wobble uridine tRNA modification.

PubMed

Chen, Changchun; Huang, Bo; Eliasson, Mattias; Rydén, Patrik; Byström, Anders S

2011-09-01

Elongator complex is required for formation of the side chains at position 5 of modified nucleosides 5-carbamoylmethyluridine (ncm⁵U₃₄), 5-methoxycarbonylmethyluridine (mcm⁵U₃₄), and 5-methoxycarbonylmethyl-2-thiouridine (mcm⁵s²U₃₄) at wobble position in tRNA. These modified nucleosides are important for efficient decoding during translation. In a recent publication, Elongator complex was implicated to participate in telomeric gene silencing and DNA damage response by interacting with proliferating cell nuclear antigen (PCNA). Here we show that elevated levels of tRNA(Lys)(s²UUU), tRNA(Gln)(s²UUG), and tRNA(Glu)(s²UUC), which in a wild-type background contain the mcm⁵s²U nucleoside at position 34, suppress the defects in telomeric gene silencing and DNA damage response observed in the Elongator mutants. We also found that the reported differences in telomeric gene silencing and DNA damage response of various elp3 alleles correlated with the levels of modified nucleosides at U₃₄. Defects in telomeric gene silencing and DNA damage response are also observed in strains with the tuc2Δ mutation, which abolish the formation of the 2-thio group of the mcm⁵s²U nucleoside in tRNA(Lys)(mcm⁵s²UUU), tRNA(Gln)(mcm⁵s²UUG), and tRNA(Glu)(mcm⁵s²UUC). These observations show that Elongator complex does not directly participate in telomeric gene silencing and DNA damage response, but rather that modified nucleosides at U₃₄ are important for efficient expression of gene products involved in these processes. Consistent with this notion, we found that expression of Sir4, a silent information regulator required for assembly of silent chromatin at telomeres, was decreased in the elp3Δ mutants.

Elongator Complex Influences Telomeric Gene Silencing and DNA Damage Response by Its Role in Wobble Uridine tRNA Modification

PubMed Central

Chen, Changchun; Huang, Bo; Eliasson, Mattias; Rydén, Patrik; Byström, Anders S.

2011-01-01

Elongator complex is required for formation of the side chains at position 5 of modified nucleosides 5-carbamoylmethyluridine (ncm5U34), 5-methoxycarbonylmethyluridine (mcm5U34), and 5-methoxycarbonylmethyl-2-thiouridine (mcm5s2U34) at wobble position in tRNA. These modified nucleosides are important for efficient decoding during translation. In a recent publication, Elongator complex was implicated to participate in telomeric gene silencing and DNA damage response by interacting with proliferating cell nuclear antigen (PCNA). Here we show that elevated levels of tRNALys s2 UUU, tRNAGln s2 UUG, and tRNAGlu s2 UUC, which in a wild-type background contain the mcm5s2U nucleoside at position 34, suppress the defects in telomeric gene silencing and DNA damage response observed in the Elongator mutants. We also found that the reported differences in telomeric gene silencing and DNA damage response of various elp3 alleles correlated with the levels of modified nucleosides at U34. Defects in telomeric gene silencing and DNA damage response are also observed in strains with the tuc2Δ mutation, which abolish the formation of the 2-thio group of the mcm5s2U nucleoside in tRNALys mcm5s2UUU, tRNAGln mcm5s2UUG, and tRNAGlu mcm5s2UUC. These observations show that Elongator complex does not directly participate in telomeric gene silencing and DNA damage response, but rather that modified nucleosides at U34 are important for efficient expression of gene products involved in these processes. Consistent with this notion, we found that expression of Sir4, a silent information regulator required for assembly of silent chromatin at telomeres, was decreased in the elp3Δ mutants. PMID:21912530

The DREO Elint Browser Utility (DEBU) reference manual

NASA Astrophysics Data System (ADS)

Ford, Barbara; Jones, David

1992-04-01

An electronic intelligent database browsing tool called DEBU has been developed that allows databases such as ELP, Kilting, EWIR, and AFEWC to be reviewed and analyzed from a user-friendly environment on a personal computer. DEBU's basic function is to allow users to examine the contents of user-selected subfiles of user-selected emitters of user-selected databases. DEBU augments this functionality with support for selecting (filtering) and combining subsets of emitters by user-selected attributes such as name, parameter type, or parameter value. DEBU provides facilities for examining histograms and x-y plots of selected parameters, for doing ambiguity analysis and mode level analysis, and for generating and printing a variety of reports. A manual is provided for users of DEBU, including descriptions and illustrations of menus and windows.

Developing Primary Propulsion for the Ares I Crew Launch Vehicle and Ares V Cargo Launch Vehicle

NASA Technical Reports Server (NTRS)

Priskos, Alex S.; Williams, Thomas L.; Ezell, Timothy G.; Burt, Rick

2007-01-01

In accordance with the U.S. Vision for Space Exploration, NASA has been tasked to send human beings to the moon, Mars, and beyond. The first stage of NASA's new Ares I crew launch vehicle (Figure 1), which will loft the Orion crew exploration vehicle into low-Earth orbit early next decade, will consist of a Space Shuttle-derived five-segment Reusable Solid Rocket Booster (RSRB); a pair of similar RSRBs also will be used on the Ares V cargo launch vehicle's core stage propulsion system. This paper will discuss the basis for choosing this particular propulsion system; describe the activities the Exploration Launch Projects (ELP) Office is engaged in at present to develop the first stage; and offer a preview of future development activities related to the first Ares l integrated test flight, which is planned for 2009.

Laser sans mode et étoile laser

NASA Astrophysics Data System (ADS)

Pique, J.-P.; Farinotti, S.; Fesquet, V.

2004-11-01

Nous décrivons la chaîne laser étudiée dans le cadre du projet Etoile-Laser-Polychromatique pour l'Optique-Adaptative (ELP-OA). Cette chaîne est constituée d'un oscillateur sans-mode, d'un préamplificateur intra-cavité et d'un amplificateur. L'objectif fixé est : 20 W de puissance moyenne, avec un taux de répétition de 15 kHz et une largeur d'impulsion de 40 ns. Nous montrons que l'aspect sans-mode de cette chaîne permet de s'affranchir du phénomène de saturation de l'atome de sodium. Nous avons mesuré un gain de 5 sur l'intensité de l'étoile monochromatique D{2}. Un facteur 30 est attendu sur l'intensité de la composante UV de l'étoile polychromatique.

Proteolytic enzyme engineering: a tool for wool.

PubMed

Araújo, Rita; Silva, Carla; Machado, Raul; Casal, Margarida; Cunha, António M; Rodriguez-Cabello, José Carlos; Cavaco-Paulo, Artur

2009-06-08

One of the goals of protein engineering is to tailor the structure of enzymes to optimize industrial bioprocesses. In the present work, we present the construction of a novel high molecular weight subtilisin, based on the fusion of the DNA sequences coding for Bacillus subtilis prosubtilisin E and for an elastin-like polymer (ELP). The resulting fusion protein was biologically produced in Escherichia coli , purified and used for wool finishing assays. When compared to the commercial protease Esperase, the recombinant subtilisinE-VPAVG(220) activity was restricted to the cuticle of wool, allowing a significant reduction of pilling, weight loss and tensile strength loss of wool fibers. Here we report, for the first time, the microbial production of a functionalized high molecular weight protease for controlled enzymatic hydrolysis of wool surface. This original process overcomes the unrestrained diffusion and extended fiber damage which are the major obstacles for the use of proteases for wool finishing applications.

Screening of potential genes contributing to the macrocycle drug resistance of C. albicans via microarray analysis

PubMed Central

Yang, Jing; Zhang, Wei; Sun, Jian; Xi, Zhiqin; Qiao, Zusha; Zhang, Jinyu; Wang, Yan; Ji, Ying; Feng, Wenli

2017-01-01

The aim of the present study was to investigate the potential genes involved in drug resistance of Candida albicans (C. albicans) by performing microarray analysis. The gene expression profile of GSE65396 was downloaded from the Gene Expression Omnibus, including a control, 15-min and 45-min macrocyclic compound RF59-treated group with three repeats for each. Following preprocessing using RAM, the differentially expressed genes (DEGs) were screened using the Limma package. Subsequently, the Kyoto Encyclopedia of Genes and Genomes pathways of these genes were analyzed using the Database for Annotation, Visualization and Integrated Discovery. Based on interactions estimated by the Search Tool for Retrieval of Interacting Gene, the protein-protein interaction (PPI) network was visualized using Cytoscape. Subnetwork analysis was performed using ReactomeFI. A total of 154 upregulated and 27 downregulated DEGs were identified in the 15-min treated group, compared with the control, and 235 upregulated and 233 downregulated DEGs were identified in the 45-min treated group, compared with the control. The upregulated DEGs were significantly enriched in the ribosome pathway. Based on the PPI network, PRP5, RCL1, NOP13, NOP4 and MRT4 were the top five nodes in the 15-min treated comparison. GIS2, URA3, NOP58, ELP3 and PLP7 were the top five nodes in the 45-min treated comparison, and its subnetwork was significantly enriched in the ribosome pathway. The macrocyclic compound RF59 had a notable effect on the ribosome and its associated pathways of C. albicans. RCL1, NOP4, MRT4, GIS2 and NOP58 may be important in RF59-resistance. PMID:28944888

Potential Interference Among The Campanian Ignimbrite Eruption, Heinrich Event 4 and The Middle/upper Palaeolithic Shift In Late Pleistocene Europe

NASA Astrophysics Data System (ADS)

Fedele, F. G.; Giaccio, B.; Isaia, R.; Orsi, G.

The Campanian Ignimbrite (CI) eruption (Phlegraean Fields Caldera) was the largest volcanic eruption in the Greater Mediterranean area over the past 200 Ka (at least 150 km3 DRE). Ash layers correlated with CI have been found in sediments of the eastern Mediterranean Sea (Y5) and East Europe, from Italy to the former USSR. The recent dating of the CI eruption at 39.3 Ka BP draws attention to the coincidence be- tween this volcanic catastrophe and the suite of coeval biocultural modifications in Old World prehistory, here termed the European Late Pleistocene shift (ELPS). These included the Middle to Upper Palaeolithic cultural transition and the supposed change from Neanderthal to "modern" Homo sapiens anatomy, still a subject of sustained de- bate. The first results of our investigations show that: (1) at several archaeological sites of peninsular Italy a distinct tephra layer corresponding to the CI is regularly interbedded between the last documented Middle Palaeolithic and the earliest appear- ance of unquestionable Upper Palaeolithic assemblages; (2) at the same sites the CI tephra coincides with a interruption of occupation, several millennia long; (3) in the GISP2 Greenland ice-core, Lago Grande di Monticchio (southern Italy) lacustrine se- quences, and KET 8003 Tyrrhenian sea-core, a large volcanogenic sulfate signal (375 ppb, at 40,062 yr BP) to be correlated with the CI eruption and/or CI tephra layer occurs just before a sharp climatic shift which coincides with the onset of Heinrich event 4 (HE4). The concurrence of the CI eruption, Palaeolithic site abandonment and beginning of HE4 suggests that the overlapping of CI eruption and HE4 climatic im- pacts induced ecosystem crisis on a fairly large scale - human systems included - and well beyond the direct-impact zone. Moreover, the occurrence of the CI eruption just before HE4 probably corroborates the positive climate-volcanism feedback supposed for other high magnitude eruptions (e.g. Toba, 74 Ka). Without obviously claiming for the CI an overall evolutionary relevance within the ELPS, on the available data we nevertheless suggest that it deserves careful consideration as a contributing factor to the regional expression - or re-orientation - of cultural and population change.

Production and characterization of in planta transiently produced polygalacturanase from Aspergillus niger and its fusions with hydrophobin or ELP tags.

PubMed

Pereira, Eridan Orlando; Kolotilin, Igor; Conley, Andrew Jonathan; Menassa, Rima

2014-06-27

Pectinases play an important role in plant cell wall deconstruction and have potential in diverse industries such as food, wine, animal feed, textile, paper, fuel, and others. The demand for such enzymes is increasing exponentially, as are the efforts to improve their production and to implement their use in several industrial processes. The goal of this study was to examine the potential of producing polygalacturonase I from Aspergillus niger in plants and to investigate the effects of subcellular compartmentalization and protein fusions on its accumulation and activity. Polygalacturonase I from Aspergillus niger (AnPGI) was transiently produced in Nicotiana benthamiana by targeting it to five different cellular compartments: apoplast, endoplasmic reticulum (ER), vacuole, chloroplast and cytosol. Accumulation levels of 2.5%, 3.0%, and 1.9% of total soluble protein (TSP) were observed in the apoplast, ER, and vacuole, respectively, and specific activity was significantly higher in vacuole-targeted AnPGI compared to the same enzyme targeted to the ER or apoplast. No accumulation was found for AnPGI when targeted to the chloroplast or cytosol. Analysis of AnPGI fused with elastin-like polypeptide (ELP) revealed a significant increase in the protein accumulation level, especially when targeted to the vacuole where the protein doubles its accumulation to 3.6% of TSP, while the hydrophobin (HFBI) fusion impaired AnPGI accumulation and both tags impaired activity, albeit to different extents. The recombinant protein showed activity against polygalacturonic acid with optimum conditions at pH 5.0 and temperature from 30 to 50°C, depending on its fusion. In vivo analysis of reducing sugar content revealed a higher release of reducing sugars in plant tissue expressing recombinant AnPGI compared to wild type N. benthamiana leaves. Our results demonstrate that subcellular compartmentalization of enzymes has an impact on both the target protein accumulation and its activity, especially in the case of proteins that undergo post-translational modifications, and should be taken into consideration when protein production strategies are designed. Using plants to produce heterologous enzymes for the degradation of a key component of the plant cell wall could reduce the cost of biomass pretreatment for the production of cellulosic biofuels.

Production and characterization of in planta transiently produced polygalacturanase from Aspergillus niger and its fusions with hydrophobin or ELP tags

PubMed Central

2014-01-01

Background Pectinases play an important role in plant cell wall deconstruction and have potential in diverse industries such as food, wine, animal feed, textile, paper, fuel, and others. The demand for such enzymes is increasing exponentially, as are the efforts to improve their production and to implement their use in several industrial processes. The goal of this study was to examine the potential of producing polygalacturonase I from Aspergillus niger in plants and to investigate the effects of subcellular compartmentalization and protein fusions on its accumulation and activity. Results Polygalacturonase I from Aspergillus niger (AnPGI) was transiently produced in Nicotiana benthamiana by targeting it to five different cellular compartments: apoplast, endoplasmic reticulum (ER), vacuole, chloroplast and cytosol. Accumulation levels of 2.5%, 3.0%, and 1.9% of total soluble protein (TSP) were observed in the apoplast, ER, and vacuole, respectively, and specific activity was significantly higher in vacuole-targeted AnPGI compared to the same enzyme targeted to the ER or apoplast. No accumulation was found for AnPGI when targeted to the chloroplast or cytosol. Analysis of AnPGI fused with elastin-like polypeptide (ELP) revealed a significant increase in the protein accumulation level, especially when targeted to the vacuole where the protein doubles its accumulation to 3.6% of TSP, while the hydrophobin (HFBI) fusion impaired AnPGI accumulation and both tags impaired activity, albeit to different extents. The recombinant protein showed activity against polygalacturonic acid with optimum conditions at pH 5.0 and temperature from 30 to 50°C, depending on its fusion. In vivo analysis of reducing sugar content revealed a higher release of reducing sugars in plant tissue expressing recombinant AnPGI compared to wild type N. benthamiana leaves. Conclusion Our results demonstrate that subcellular compartmentalization of enzymes has an impact on both the target protein accumulation and its activity, especially in the case of proteins that undergo post-translational modifications, and should be taken into consideration when protein production strategies are designed. Using plants to produce heterologous enzymes for the degradation of a key component of the plant cell wall could reduce the cost of biomass pretreatment for the production of cellulosic biofuels. PMID:24970673

Single nucleotide polymorphisms for feed efficiency and performance in crossbred beef cattle

PubMed Central

2014-01-01

Background This study was conducted to: (1) identify new SNPs for residual feed intake (RFI) and performance traits within candidate genes identified in a genome wide association study (GWAS); (2) estimate the proportion of variation in RFI explained by the detected SNPs; (3) estimate the effects of detected SNPs on carcass traits to avoid undesirable correlated effects on these economically important traits when selecting for feed efficiency; and (4) map the genes to biological mechanisms and pathways. A total number of 339 SNPs corresponding to 180 genes were tested for association with phenotypes using a single locus regression (SLRM) and genotypic model on 726 and 990 crossbred animals for feed efficiency and carcass traits, respectively. Results Strong evidence of associations for RFI were located on chromosomes 8, 15, 16, 18, 19, 21, and 28. The strongest association with RFI (P = 0.0017) was found with a newly discovered SNP located on BTA 8 within the ELP3 gene. SNPs rs41820824 and rs41821600 on BTA 16 within the gene HMCN1 were strongly associated with RFI (P = 0.0064 and P = 0.0033, respectively). A SNP located on BTA 18 within the ZNF423 gene provided strong evidence for association with RFI (P = 0.0028). Genomic estimated breeding values (GEBV) from 98 significant SNPs were moderately correlated (0.47) to the estimated breeding values (EBVs) from a mixed animal model. The significant (P < 0.05) SNPs (98) explained 26% of the genetic variance for RFI. In silico functional analysis for the genes suggested 35 and 39 biological processes and pathways, respectively for feed efficiency traits. Conclusions This study identified several positional and functional candidate genes involved in important biological mechanisms associated with feed efficiency and performance. Significant SNPs should be validated in other populations to establish their potential utilization in genetic improvement programs. PMID:24476087

Low-Tech, Pilot Scale Purification of a Recombinant Spider Silk Protein Analog from Tobacco Leaves.

PubMed

Heppner, René; Weichert, Nicola; Schierhorn, Angelika; Conrad, Udo; Pietzsch, Markus

2016-10-09

Spider dragline is used by many members of the Araneae family not only as a proteinogenic safety thread but also for web construction. Spider dragline has been shown to possess high tensile strength in combination with elastic behavior. This high tensile strength can be attributed to the presence of antiparallel β-sheets within the thread; these antiparallel β-sheets are why the protein is classified as a silk. Due to the properties of spider silk and its technical and medical uses, including its use as a suture material and as a scaffold for tissue regeneration, spider dragline is a focus of the biotechnology industry. The production of sufficient amounts of spider silk is challenging, as it is difficult to produce large quantities of fibers because of the cannibalistic behavior of spiders and their large spatial requirements. In recent years, the heterologous expression of genes coding for spider silk analogs in various hosts, including plants such as Nicotiana tabacum , has been established. We developed a simple and scalable method for the purification of a recombinant spider silk protein elastin-like peptide fusion protein (Q-/K-MaSp1-100× ELP) after heterologous production in tobacco leaves involving heat and acetone precipitation. Further purification was performed using centrifugal Inverse Transition Cycling (cITC). Up to 400 mg of highly pure spider silk protein derivatives can be isolated from six kilograms of tobacco leaves, which is the highest amount of silk protein derivatives purified from plants thus far.

Expression and purification of moricin CM4 and human β-defensins 4 in Escherichia coli using a new technology.

PubMed

Shen, Yang; Ai, Hong-Xin; Song, Ren; Liang, Zhen-Ning; Li, Jian-Feng; Zhang, Shuang-Quan

2010-10-20

Different strategies have been developed to produce small antimicrobial peptides using recombinant techniques. Here we report a new technology of biosynthesis of moricin CM4 and human β-defensins 4 (HβD4) in the Escherichia coli. The CM4 and HβD4 gene were cloned into a vector containing the tags elastin-like peptide (ELP) and intein to construct the expression vector pET-EI-CM4 and pET-EI-HβD4. All the peptides, expressed as soluble fusions, were isolated from the protein debris by the method called inverse transition cycling (ITC) rather than traditional immobilized metal affinity chromatography (IMAC) and separated from the fusion leader by self-cleavage. Fully reduced peptides that were purified exhibited expected antimicrobial activity. The approach described here is a low-cost, convenient and potential way for generating small antimicrobial peptide. Copyright © 2010 Elsevier GmbH. All rights reserved.

Efficacy of reducing agent and surfactant contacting pattern on the performance characteristics of nickel electroless plating baths coupled with and without ultrasound.

PubMed

Agarwal, Amrita; Pujari, Murali; Uppaluri, Ramgopal; Verma, Anil

2014-07-01

This article addresses furthering the role of sonication for the optimal fabrication of nickel ceramic composite membranes using electroless plating. Deliberating upon process modifications for surfactant induced electroless plating (SIEP) and combined surfactant and sonication induced electroless plating (SSOEP), this article highlights a novel method of contacting of the reducing agent and surfactant to the conventional electroless nickel plating baths. Rigorous experimental investigations indicated that the combination of ultrasound (in degas mode), surfactant and reducing agent pattern had a profound influence in altering the combinatorial plating characteristics. For comparison purpose, purely surfactant induced nickel ELP baths have also been investigated. These novel insights consolidate newer research horizons for the role of ultrasound to achieve dense metal ceramic composite membranes in a shorter span of total plating time. Surface and physical characterizations were carried out using BET, FTIR, XRD, FESEM and nitrogen permeation experiments. It has been analyzed that the SSOEP baths provided maximum ratio of percent pore densification per unit metal film thickness (PPDδ) and hold the key for further fine tuning of the associated degrees of freedom. On the other hand SIEP baths provided lower (PPDδ) ratio but higher PPD. For SSOEP baths with dropwise reducing agent and bulk surfactant, the PPD and metal film thickness values were 73.4% and 8.4 μm which varied to 66.9% and 13.3 μm for dropwise reducing agent and drop surfactant case. Copyright © 2014 Elsevier B.V. All rights reserved.

Nematic director reorientation at solid and liquid interfaces under flow: SAXS studies in a microfluidic device

DOE PAGES

Silva, Bruno F. B.; Zepeda-Rosales, Miguel; Venkateswaran, Neeraja; ...

2014-10-30

In this work we investigate the interplay between flow and boundary condition effects on the orientation field of a thermotropic nematic liquid crystal under flow and confinement in a microfluidic device. Two types of experiments were performed using synchrotron small-angle X-ray-scattering (SAXS). In the first, a nematic liquid crystal flows through a square-channel cross section at varying flow rates, while the nematic director orientation projected onto the velocity/velocity gradient plane is measured using a 2D detector. At moderate-to-high flow rates, the nematic director is predominantly aligned in the flow direction, but with a small tilt angle of ~±11° in themore » velocity gradient direction. The director tilt angle is constant throughout most of the channel width but switches sign when crossing the center of the channel, in agreement with the Ericksen–Leslie–Parodi (ELP) theory. At low flow rates, boundary conditions begin to dominate, and a flow profile resembling the escaped radial director configuration is observed, where the director is seen to vary more smoothly from the edges (with homeotropic alignment) to the center of the channel. In the second experiment, hydrodynamic focusing is employed to confine the nematic phase into a sheet of liquid sandwiched between two layers of Triton X-100 aqueous solutions. The average nematic director orientation shifts to some extent from the flow direction toward the liquid boundaries, although it remains unclear if one tilt angle is dominant through most of the nematic sheet (with abrupt jumps near the boundaries) or if the tilt angle varies smoothly between two extreme values (~90 and 0°). Lastly, the technique presented here could be applied to perform high-throughput measurements for assessing the influence of different surfactants on the orientation of nematic phases and may lead to further improvements in areas such as boundary lubrication and clarifying the nature of defect structures in LC displays.« less

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

Silva, Bruno F. B.; Zepeda-Rosales, Miguel; Venkateswaran, Neeraja

In this work we investigate the interplay between flow and boundary condition effects on the orientation field of a thermotropic nematic liquid crystal under flow and confinement in a microfluidic device. Two types of experiments were performed using synchrotron small-angle X-ray-scattering (SAXS). In the first, a nematic liquid crystal flows through a square-channel cross section at varying flow rates, while the nematic director orientation projected onto the velocity/velocity gradient plane is measured using a 2D detector. At moderate-to-high flow rates, the nematic director is predominantly aligned in the flow direction, but with a small tilt angle of ~±11° in themore » velocity gradient direction. The director tilt angle is constant throughout most of the channel width but switches sign when crossing the center of the channel, in agreement with the Ericksen–Leslie–Parodi (ELP) theory. At low flow rates, boundary conditions begin to dominate, and a flow profile resembling the escaped radial director configuration is observed, where the director is seen to vary more smoothly from the edges (with homeotropic alignment) to the center of the channel. In the second experiment, hydrodynamic focusing is employed to confine the nematic phase into a sheet of liquid sandwiched between two layers of Triton X-100 aqueous solutions. The average nematic director orientation shifts to some extent from the flow direction toward the liquid boundaries, although it remains unclear if one tilt angle is dominant through most of the nematic sheet (with abrupt jumps near the boundaries) or if the tilt angle varies smoothly between two extreme values (~90 and 0°). Lastly, the technique presented here could be applied to perform high-throughput measurements for assessing the influence of different surfactants on the orientation of nematic phases and may lead to further improvements in areas such as boundary lubrication and clarifying the nature of defect structures in LC displays.« less

Isolation of autochthonous non-white rot fungi with potential for enzymatic upgrading of Venezuelan extra-heavy crude oil

PubMed Central

Naranjo, Leopoldo; Urbina, Hector; De Sisto, Angela; Leon, Vladimir

2007-01-01

The increasing world demand for fuels makes it necessary to exploit the largest reserve of extra-heavy crude oil (EHCO) of the Orinoco Oil Belt from Venezuela. We propose the use of extracellular oxidative enzymes, in particular, lignin-degrading enzyme systems (LDS) of fungi, for enzymatic improvement of EHCO. Autochthonous non-white rot fungal strains able to use EHCO, and several polycyclic aromatic hydrocarbons (PAHs) as sole carbon source and energy, were isolated from EHCO-polluted soils and identified as belonging to the genera Fusarium, Penicillium , Trichoderma , Aspergillus , Neosartorya, Pseudallescheria, Cladosporium, Pestalotiopsis , Phoma and Paecillomyces. Phenotypic and biochemical assays revealed the ability of these filamentous fungi to synthesize extracellular oxidative enzymes, and suggested a relationship between the LDS and EHCO bioconversion. This work reports, for the first time, the use of o-phenylenediamine dihydrochloride (OPD) as substrate to measure extracellular ligninolytic peroxidases (ELP) in culture broths of filamentous fungi (Fusarium solani HP-1), and constitutes the first formal study of the fungal community associated with the EHCO of the Orinoco Oil Belt. PMID:18833334

Current Treatments in Familial Dysautonomia

PubMed Central

Palma, Jose-Alberto; Kaufmann, Lucy; Fuente, Cristina; Percival, Leila; Mendoza, Carlos; Kaufmann, Horacio

2014-01-01

Introduction Familial dysautonomia (FD) is a rare hereditary sensory and autonomic neuropathy (type III). The disease is caused by a point mutation in the IKBKAP gene that affects the splicing of the elongator-1 protein (also known as IKAP). Patients have dramatic blood pressure instability due to baroreflex failure, chronic kidney disease, and impaired swallowing leading to recurrent aspiration pneumonia, which results in chronic lung disease. Diminished pain and temperature perception results in neuropathic joints and thermal injuries. Impaired proprioception leads to gait ataxia. Optic neuropathy and corneal opacities lead to progressive visual loss. Areas covered This article reviews current therapeutic strategies for the symptomatic treatment of FD, as well as the potential of new gene modifying agents. Expert opinion Therapeutic focus on FD is centered on reducing the catecholamine surges caused by baroreflex failure. Managing neurogenic dysphagia with effective protection of the airway passages and prompt treatment of aspiration pneumonias is necessary to prevent respiratory failure. Sedative medications should be used cautiously due to risk of respiratory depression. Non-invasive ventilation during sleep effectively manages apneas and prevents hypercapnia. Clinical trials of compounds that increase levels of IKAP (ELP-1) are underway and will determine whether they can reverse or slow disease progression. PMID:25323828

The impairment of learning and memory and synaptic loss in mouse after chronic nitrite exposure.

PubMed

Chen, Yongfang; Cui, Zhanjun; Wang, Lai; Liu, Hongliang; Fan, Wenjuan; Deng, Jinbo; Deng, Jiexin

2016-12-01

The objective of this study is to understand the impairment of learning and memory in mouse after chronic nitrite exposure. The animal model of nitrite exposure in mouse was created with the daily intubation of nitrite in adult healthy male mice for 3 months. Furthermore, the mouse's learning and memory abilities were tested with Morris water maze, and the expression of Synaptophysin and γ-Synuclein was visualized with immunocytochemistry and Western blot. Our results showed that nitrite exposure significantly prolonged the escape latency period (ELP) and decreased the values of the frequency across platform (FAP) as well as the accumulative time in target quadrant (ATITQ) compared to control, in dose-dependent manner. In addition, after nitrite exposure, synaptophysin (SYN) positive buttons in the visual cortex was reduced, in contrast the increase of γ-synuclein positive cells. The results above were supported by Western blot as well. We conclude that nitrite exposure could lead to a decline in mice's learning and memory. The overexpression of γ-synuclein contributed to the synaptic loss, which is most likely the cause of learning and memory impairment. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1720-1730, 2016. © 2015 Wiley Periodicals, Inc.

Preparation and mechanical properties of layers made of recombinant spider silk proteins and silk from silk worm

NASA Astrophysics Data System (ADS)

Junghans, F.; Morawietz, M.; Conrad, U.; Scheibel, T.; Heilmann, A.; Spohn, U.

2006-02-01

Layers of recombinant spider silks and native silks from silk worms were prepared by spin-coating and casting of various solutions. FT-IR spectra were recorded to investigate the influence of the different mechanical stress occurring during the preparation of the silk layers. The solubility of the recombinant spider silk proteins SO1-ELP, C16, AQ24NR3, and of the silk fibroin from Bombyx mori were investigated in hexafluorisopropanol, ionic liquids and concentrated salt solutions. The morphology and thickness of the layers were determined by Atomic Force Microscopy (AFM) or with a profilometer. The mechanical behaviour was investigated by acoustic impedance analysis by using a quartz crystal microbalance (QCMB) as well as by microindentation. The density of silk layers (d<300 nm) was determined based on AFM and QCMB measurements. At silk layers thicker than 300 nm significant changes of the half-band-half width can be correlated with increasing energy dissipation. Microhardness measurements demonstrate that recombinant spider silk and sericine-free Bombyx mori silk layers achieve higher elastic penetration modules EEP and Martens hardness values HM than those of polyethylenterephthalate (PET) and polyetherimide (PEI) foils.

Separation efficiency of free-solution conjugated electrophoresis with drag-tags incorporating a synthetic amino acid.

PubMed

Seo, Kyung-Ho; Chu, Hun-Su; Yoo, Tae Hyeon; Lee, Sun-Gu; Won, Jong-In

2016-03-01

DNA sequencing or separation by conventional capillary electrophoresis with a polymer matrix has some inherent drawbacks, such as the expense of polymer matrix and limitations in sequencing read length. As DNA fragments have a linear charge-to-friction ratio in free solution, DNA fragments cannot be separated by size. However, size-based separation of DNA is possible in free-solution conjugate electrophoresis (FSCE) if a "drag-tag" is attached to DNA fragments because the tag breaks the linear charge-to-friction scaling. Although several previous studies have demonstrated the feasibility of DNA separation by free-solution conjugated electrophoresis, generation of a monodisperse drag-tag and identification of a strong, site-specific conjugation method between a DNA fragment and a drag-tag are challenges that still remain. In this study, we demonstrate an efficient FSCE method by conjugating a biologically synthesized elastin-like polypeptide (ELP) and green fluorescent protein (GFP) to DNA fragments. In addition, to produce strong and site-specific conjugation, a methionine residue in drag-tags is replaced with homopropargylglycine (Hpg), which can be conjugated specifically to a DNA fragment with an azide site. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

30 CFR 815.15 - Performance standards for coal exploration.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 30 Mineral Resources 3 2012-07-01 2012-07-01 false Performance standards for coal exploration. 815... OF THE INTERIOR PERMANENT PROGRAM PERFORMANCE STANDARDS PERMANENT PROGRAM PERFORMANCE STANDARDS-COAL EXPLORATION § 815.15 Performance standards for coal exploration. (a) Habitats of unique or unusually high...

30 CFR 815.15 - Performance standards for coal exploration.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 30 Mineral Resources 3 2010-07-01 2010-07-01 false Performance standards for coal exploration. 815... OF THE INTERIOR PERMANENT PROGRAM PERFORMANCE STANDARDS PERMANENT PROGRAM PERFORMANCE STANDARDS-COAL EXPLORATION § 815.15 Performance standards for coal exploration. (a) Habitats of unique or unusually high...

30 CFR 815.15 - Performance standards for coal exploration.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 30 Mineral Resources 3 2013-07-01 2013-07-01 false Performance standards for coal exploration. 815... OF THE INTERIOR PERMANENT PROGRAM PERFORMANCE STANDARDS PERMANENT PROGRAM PERFORMANCE STANDARDS-COAL EXPLORATION § 815.15 Performance standards for coal exploration. (a) Habitats of unique or unusually high...

30 CFR 815.15 - Performance standards for coal exploration.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 30 Mineral Resources 3 2014-07-01 2014-07-01 false Performance standards for coal exploration. 815... OF THE INTERIOR PERMANENT PROGRAM PERFORMANCE STANDARDS PERMANENT PROGRAM PERFORMANCE STANDARDS-COAL EXPLORATION § 815.15 Performance standards for coal exploration. (a) Habitats of unique or unusually high...

30 CFR 815.15 - Performance standards for coal exploration.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 30 Mineral Resources 3 2011-07-01 2011-07-01 false Performance standards for coal exploration. 815... OF THE INTERIOR PERMANENT PROGRAM PERFORMANCE STANDARDS PERMANENT PROGRAM PERFORMANCE STANDARDS-COAL EXPLORATION § 815.15 Performance standards for coal exploration. (a) Habitats of unique or unusually high...

40 CFR 434.55 - New source performance standards (NSPS).

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 29 2010-07-01 2010-07-01 false New source performance standards (NSPS... PERFORMANCE STANDARDS Post-Mining Areas § 434.55 New source performance standards (NSPS). The following new source performance standards shall apply to the post-mining areas of all new source coal mines: (a...

Aiming High: Setting Performance Standards for Student Success

ERIC Educational Resources Information Center

Phillips, Gary; Garcia, Alicia N.

2015-01-01

Content standards, not performance standards, have been almost the sole focus of state policies and recent conversations about academic standards. Without rigorous content and performance standards, we cannot adequately prepare students for the global marketplace. A recent AIR study shows that state performance standards are consistently low and…

48 CFR 37.603 - Performance standards.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 48 Federal Acquisition Regulations System 1 2012-10-01 2012-10-01 false Performance standards. 37... CATEGORIES OF CONTRACTING SERVICE CONTRACTING Performance-Based Acquisition 37.603 Performance standards. (a) Performance standards establish the performance level required by the Government to meet the contract...

48 CFR 37.603 - Performance standards.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 48 Federal Acquisition Regulations System 1 2014-10-01 2014-10-01 false Performance standards. 37... CATEGORIES OF CONTRACTING SERVICE CONTRACTING Performance-Based Acquisition 37.603 Performance standards. (a) Performance standards establish the performance level required by the Government to meet the contract...

48 CFR 37.603 - Performance standards.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 48 Federal Acquisition Regulations System 1 2010-10-01 2010-10-01 false Performance standards. 37... CATEGORIES OF CONTRACTING SERVICE CONTRACTING Performance-Based Acquisition 37.603 Performance standards. (a) Performance standards establish the performance level required by the Government to meet the contract...

48 CFR 37.603 - Performance standards.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 48 Federal Acquisition Regulations System 1 2011-10-01 2011-10-01 false Performance standards. 37... CATEGORIES OF CONTRACTING SERVICE CONTRACTING Performance-Based Acquisition 37.603 Performance standards. (a) Performance standards establish the performance level required by the Government to meet the contract...

The Relevance of Using the Moon's Age as AN Alternative in Imkanur Rukyah Criteria

NASA Astrophysics Data System (ADS)

Anwar, S.; Omar, K. M.; Che Awang, M. S.

2016-09-01

The Imkanur rukyah criteria can be defined as the minimum limit in expecting the new moon's visibility in determining the beginning of Hijri months. It has been used in the development of Hijri calendar in Malaysia since 1992. Based on the criteria, the new moon is considered visible if the altitude at sunset is at least 2° and the elongation between the moon and the sun is at least 3°, or at moonset, the age of the moon is at least 8 hours. The altitude limit of 2° and the elongation limit of 3° indeed were determined according to the data of new moon visibility observed in Indonesia, whereas for the 8-hour moon's age, there is no written rule regarding to it. The use of the moon's age criterion as an alternative to the geometric criteria can lead to confusion if both conditions provide different results. Therefore, this study was performed to assess the relevance of using moon's age as an alternative in Imkanur rukyah criteria used in Malaysia. The study utilised the data of the sun and the moon's positions, the time of sunset, the time of moonset and the time of conjunction (new moon). The data for the sun were calculated based on VSOP87 theory, while for the moon, using ELP2000-82b. Based on the analysis, in determining Hijri dates from 1996 to 2015, there are 22 discrepancies found between the moon's age and the geometric criteria, in which, 5 of them occur in the month of Ramadan, Syawal and Zulhijjah. These conditions show that the moon's age criterion is not always consistent with the geometric criteria. Therefore, the use of moon's age as an alternate criterion in determining the beginning of Hijri month is considered irrelevant and should be further reviewed.

20 CFR 627.470 - Performance standards.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false Performance standards. 627.470 Section 627... GOVERNING PROGRAMS UNDER TITLES I, II, AND III OF THE ACT Administrative Standards § 627.470 Performance standards. (a) General. The Secretary shall prescribe performance standards for adult programs under title...

20 CFR 627.470 - Performance standards.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 20 Employees' Benefits 3 2012-04-01 2012-04-01 false Performance standards. 627.470 Section 627... GOVERNING PROGRAMS UNDER TITLES I, II, AND III OF THE ACT Administrative Standards § 627.470 Performance standards. (a) General. The Secretary shall prescribe performance standards for adult programs under title...

Numerical Study of Charged Inertial Particles in Turbulence using a Coupled Fluid-P3M Approach

NASA Astrophysics Data System (ADS)

Yao, Yuan; Capecelatro, Jesse

2017-11-01

Non-trivial interactions between charged particles and turbulence play an important role in many engineering and environmental flows, including clouds, fluidized bed reactors, charged hydrocarbon sprays and dusty plasmas. Due to the long-range nature of electrostatic forces, Coulomb interactions in systems with many particles must be handled carefully to avoid O(N2) computations. The particle-mesh (PM) method is typically employed in Eulerian-Lagrangian (EL) simulations as it avoids computing direct pairwise sums, but it fails to capture short-range interactions that are anticipated to be important when particles cluster. In this presentation, the particle-particle-particle-mesh (P3M) method that scales with O(NlogN) is implemented within a EL framework to simulate charged particles accurately in a tractable manner. The EL-P3M method is used to assess the competition between drag and Coulomb forces for a range of Stokes numbers and charges. Simulations of like- and oppositely-charged particles suspended in a two-dimensional Taylor-Green vortex and three-dimensional homogeneous isotropic turbulence are reported. One-point and two-point statistics obtained using PM and P3M are compared to assess the effect of added accuracy on collision rate and clustering.

Mutation of a chitinase-like gene causes ectopic deposition of lignin, aberrant cell shapes, and overproduction of ethylene.

PubMed

Zhong, Ruiqin; Kays, Stanley J; Schroeder, Betty P; Ye, Zheng-Hua

2002-01-01

Chitinase-like proteins have long been proposed to play roles in normal plant growth and development, but no mutations in chitinase-like genes have been obtained previously to support this hypothesis. In this study, we have shown that the gene responsible for the elp1 mutation in Arabidopsis encodes a chitinase-like protein (AtCTL1). Mutation of this chitinase-like gene caused ectopic deposition of lignin and aberrant shapes of cells with incomplete cell walls in the pith of inflorescence stems. The AtCTL1 gene was expressed in all organs during normal plant growth and development, but it was not induced by wounding, salicylic acid, pectin fragments, or ethylene. Consistent with its ubiquitous expression pattern, mutation of the AtCTL1 gene affected many aspects of plant growth and development, including exaggerated hook curvature, reduced length and increased diameter of hypocotyls in dark-grown seedlings, and reduced root length and increased number of root hairs in light-grown seedlings. The mutant phenotypes could be rescued partially by ethylene inhibitors, and ethylene production in the mutant was significantly greater than in the wild type. Together, these results suggest that AtCTL1, a chitinase-like gene, is essential for normal plant growth and development in Arabidopsis.

Kinetin improves IKBKAP mRNA splicing in patients with familial dysautonomia

PubMed Central

Axelrod, Felicia B.; Liebes, Leonard; Gold-von Simson, Gabrielle; Mendoza, Sandra; Mull, James; Leyne, Maire; Norcliffe-Kaufmann, Lucy; Kaufmann, Horacio; Slaugenhaupt, Susan A.

2011-01-01

Familial dysautonomia (FD) is caused by an intronic splice mutation in the IKBKAP gene that leads to partial skipping of exon 20 and tissue-specific reduction in I-κ-B kinase complex associated protein/ elongation protein 1 (IKAP/ELP-1) expression. Kinetin (6-furfurylaminopurine) has been shown to improve splicing and increase wild-type IKBKAP mRNA and IKAP protein expression in FD cell lines and carriers. To determine if oral kinetin treatment could alter mRNA splicing in FD subjects and was tolerable, we administered kinetin to eight FD individuals homozygous for the splice mutation. Subjects received 23.5 mg/Kg/day for 28 days. An increase in wild-type IKBKAP mRNA expression in leukocytes was noted after eight days in six of eight individuals; after 28 days the mean increase as compared to baseline was significant (p=0.002). We have demonstrated that kinetin is tolerable in this medically fragile population. Not only did kinetin produce the desired effect on splicing in FD patients, but also that effect appears to improve with time despite lack of dose change. This is the first report of a drug that produces in vivo mRNA splicing changes in individuals with FD and supports future long-term trials to determine if kinetin will prove therapeutic in FD patients. PMID:21775922

40 CFR 428.15 - Standards of performance for new sources.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 29 2010-07-01 2010-07-01 false Standards of performance for new...) EFFLUENT GUIDELINES AND STANDARDS RUBBER MANUFACTURING POINT SOURCE CATEGORY Tire and Inner Tube Plants Subcategory § 428.15 Standards of performance for new sources. The following standards of performance...

12 CFR 563e.26 - Small savings association performance standards.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 12 Banks and Banking 5 2010-01-01 2010-01-01 false Small savings association performance standards... COMMUNITY REINVESTMENT Standards for Assessing Performance § 563e.26 Small savings association performance standards. (a) Performance criteria—(1) Small savings associations that are not intermediate small savings...

12 CFR 228.26 - Small bank performance standards.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 12 Banks and Banking 3 2011-01-01 2011-01-01 false Small bank performance standards. 228.26... RESERVE SYSTEM COMMUNITY REINVESTMENT (REGULATION BB) Standards for Assessing Performance § 228.26 Small bank performance standards. (a) Performance criteria—(1) Small banks that are not intermediate small...

12 CFR 563e.26 - Small savings association performance standards.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 12 Banks and Banking 5 2011-01-01 2011-01-01 false Small savings association performance standards... COMMUNITY REINVESTMENT Standards for Assessing Performance § 563e.26 Small savings association performance standards. (a) Performance criteria—(1) Small savings associations that are not intermediate small savings...

12 CFR 25.26 - Small bank performance standards.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 12 Banks and Banking 1 2011-01-01 2011-01-01 false Small bank performance standards. 25.26 Section... REINVESTMENT ACT AND INTERSTATE DEPOSIT PRODUCTION REGULATIONS Regulations Standards for Assessing Performance § 25.26 Small bank performance standards. (a) Performance criteria—(1) Small banks that are not...

12 CFR 345.26 - Small bank performance standards.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 12 Banks and Banking 4 2011-01-01 2011-01-01 false Small bank performance standards. 345.26... GENERAL POLICY COMMUNITY REINVESTMENT Standards for Assessing Performance § 345.26 Small bank performance standards. (a) Performance criteria—(1) Small banks that are not intermediate small banks. The FDIC...

12 CFR 345.26 - Small bank performance standards.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 12 Banks and Banking 4 2010-01-01 2010-01-01 false Small bank performance standards. 345.26... GENERAL POLICY COMMUNITY REINVESTMENT Standards for Assessing Performance § 345.26 Small bank performance standards. (a) Performance criteria—(1) Small banks that are not intermediate small banks. The FDIC...

12 CFR 228.26 - Small bank performance standards.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 12 Banks and Banking 3 2010-01-01 2010-01-01 false Small bank performance standards. 228.26... RESERVE SYSTEM COMMUNITY REINVESTMENT (REGULATION BB) Standards for Assessing Performance § 228.26 Small bank performance standards. (a) Performance criteria—(1) Small banks that are not intermediate small...

12 CFR 25.26 - Small bank performance standards.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 12 Banks and Banking 1 2010-01-01 2010-01-01 false Small bank performance standards. 25.26 Section... REINVESTMENT ACT AND INTERSTATE DEPOSIT PRODUCTION REGULATIONS Regulations Standards for Assessing Performance § 25.26 Small bank performance standards. (a) Performance criteria—(1) Small banks that are not...

19 CFR 143.6 - Failure to maintain performance standards.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 19 Customs Duties 2 2010-04-01 2010-04-01 false Failure to maintain performance standards. 143.6... maintain performance standards. ABI participants must adhere to the performance requirements and... maximum of 90 days, if the participant's performance remains below standard, but, except for immediate...

32 CFR 1656.11 - Job performance standards and sanctions.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 32 National Defense 6 2013-07-01 2013-07-01 false Job performance standards and sanctions. 1656.11... ALTERNATIVE SERVICE § 1656.11 Job performance standards and sanctions. (a) Standards of Performance. An ASW is... employer of his other employees in similar jobs. If there are no other employees, the standards shall...

32 CFR 1656.11 - Job performance standards and sanctions.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 32 National Defense 6 2011-07-01 2011-07-01 false Job performance standards and sanctions. 1656.11... ALTERNATIVE SERVICE § 1656.11 Job performance standards and sanctions. (a) Standards of Performance. An ASW is... employer of his other employees in similar jobs. If there are no other employees, the standards shall...

32 CFR 1656.11 - Job performance standards and sanctions.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 32 National Defense 6 2012-07-01 2012-07-01 false Job performance standards and sanctions. 1656.11... ALTERNATIVE SERVICE § 1656.11 Job performance standards and sanctions. (a) Standards of Performance. An ASW is... employer of his other employees in similar jobs. If there are no other employees, the standards shall...

32 CFR 1656.11 - Job performance standards and sanctions.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 32 National Defense 6 2014-07-01 2014-07-01 false Job performance standards and sanctions. 1656.11... ALTERNATIVE SERVICE § 1656.11 Job performance standards and sanctions. (a) Standards of Performance. An ASW is... employer of his other employees in similar jobs. If there are no other employees, the standards shall...

2006 Hawaii State Performance Standards Review Commission: "Final Report"

ERIC Educational Resources Information Center

State of Hawaii Department of Education, 2006

2006-01-01

The 2006 Performance Standards Review Commission (PSRC) was appointed by the Board of Education "to assess the effectiveness of the performance standards and to review the implementation of the performance standards by the Board and the schools to determine whether the standards should be modified" (Section 302A-201, Hawaii Revised…

Self-Assembled Materials Made from Functional Recombinant Proteins.

PubMed

Jang, Yeongseon; Champion, Julie A

2016-10-18

Proteins are potent molecules that can be used as therapeutics, sensors, and biocatalysts with many advantages over small-molecule counterparts due to the specificity of their activity based on their amino acid sequence and folded three-dimensional structure. However, they also have significant limitations in their stability, localization, and recovery when used in soluble form. These opportunities and challenges have motivated the creation of materials from such functional proteins in order to protect and present them in a way that enhances their function. We have designed functional recombinant fusion proteins capable of self-assembling into materials with unique structures that maintain or improve the functionality of the protein. Fusion of either a functional protein or an assembly domain to a leucine zipper domain makes the materials design strategy modular, based on the high affinity between leucine zippers. The self-assembly domains, including elastin-like polypeptides (ELPs) and defined-sequence random coil polypeptides, can be fused with a leucine zipper motif in order to promote assembly of the fusion proteins into larger structures upon specific stimuli such as temperature and ionic strength. Fusion of other functional domains with the counterpart leucine zipper motif endows the self-assembled materials with protein-specific functions such as fluorescence or catalytic activity. In this Account, we describe several examples of materials assembled from functional fusion proteins as well as the structural characterization, functionality, and understanding of the assembly mechanism. The first example is zipper fusion proteins containing ELPs that assemble into particles when introduced to a model extracellular matrix and subsequently disassemble over time to release the functional protein for drug delivery applications. Under different conditions, the same fusion proteins can self-assemble into hollow vesicles. The vesicles display a functional protein on the surface and can also carry protein, small-molecule, or nanoparticle cargo in the vesicle lumen. To create a material with a more complex hierarchical structure, we combined calcium phosphate with zipper fusion proteins containing random coil polypeptides to produce hybrid protein-inorganic supraparticles with high surface area and porous structure. The use of a functional enzyme created supraparticles with the ability to degrade inflammatory cytokines. Our characterization of these protein materials revealed that the molecular interactions are complex because of the large size of the protein building blocks, their folded structures, and the number of potential interactions including hydrophobic interactions, electrostatic interactions, van der Waals forces, and specific affinity-based interactions. It is difficult or even impossible to predict the structures a priori. However, once the basic assembly principles are understood, there is opportunity to tune the material properties, such as size, through control of the self-assembly conditions. Our future efforts on the fundamental side will focus on identifying the phase space of self-assembly of these fusion proteins and additional experimental levers with which to control and tune the resulting materials. On the application side, we are investigating an array of different functional proteins to expand the use of these structures in both therapeutic protein delivery and biocatalysis.

42 CFR 421.122 - Performance standards.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 42 Public Health 3 2013-10-01 2013-10-01 false Performance standards. 421.122 Section 421.122... (CONTINUED) MEDICARE PROGRAM (CONTINUED) MEDICARE CONTRACTING Intermediaries § 421.122 Performance standards. (a) Development of standards. In addition to the performance criteria (§ 421.120), CMS develops...

42 CFR 421.122 - Performance standards.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 42 Public Health 3 2012-10-01 2012-10-01 false Performance standards. 421.122 Section 421.122... (CONTINUED) MEDICARE PROGRAM (CONTINUED) MEDICARE CONTRACTING Intermediaries § 421.122 Performance standards. (a) Development of standards. In addition to the performance criteria (§ 421.120), CMS develops...

42 CFR 421.122 - Performance standards.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 42 Public Health 3 2011-10-01 2011-10-01 false Performance standards. 421.122 Section 421.122... (CONTINUED) MEDICARE PROGRAM MEDICARE CONTRACTING Intermediaries § 421.122 Performance standards. (a) Development of standards. In addition to the performance criteria (§ 421.120), CMS develops detailed...

24 CFR 3400.113 - Performance standards.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 24 Housing and Urban Development 5 2014-04-01 2014-04-01 false Performance standards. 3400.113....113 Performance standards. (a) For HUD to determine that a state is providing effective supervision and enforcement, a supervisory authority must meet the following performance standards: (1) The...

24 CFR 3400.113 - Performance standards.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 24 Housing and Urban Development 5 2013-04-01 2013-04-01 false Performance standards. 3400.113....113 Performance standards. (a) For HUD to determine that a state is providing effective supervision and enforcement, a supervisory authority must meet the following performance standards: (1) The...

42 CFR 421.122 - Performance standards.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 42 Public Health 3 2014-10-01 2014-10-01 false Performance standards. 421.122 Section 421.122... (CONTINUED) MEDICARE PROGRAM (CONTINUED) MEDICARE CONTRACTING Intermediaries § 421.122 Performance standards. (a) Development of standards. In addition to the performance criteria (§ 421.120), CMS develops...

75 FR 20671 - Seventieth Meeting: RTCA Special Committee 147: Minimum Operational Performance Standards for...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-04-20

... Committee 147: Minimum Operational Performance Standards for Traffic Alert and Collision Avoidance Systems... Committee 147: Minimum Operational Performance Standards for Traffic Alert and Collision Avoidance Systems... RTCA Special Committee 147: Minimum Operational Performance Standards for Traffic Alert and Collision...

Setting Emissions Standards Based on Technology Performance

EPA Pesticide Factsheets

In setting national emissions standards, EPA sets emissions performance levels rather than mandating use of a particular technology. The law mandates that EPA use numerical performance standards whenever feasible in setting national emissions standards.

76 FR 11846 - Seventy-second Meeting: RTCA Special Committee 147: Minimum Operational Performance Standards for...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-03-03

... Committee 147: Minimum Operational Performance Standards for Traffic Alert and Collision Avoidance Systems... Committee 147 meeting: Minimum Operational Performance Standards for Traffic Alert and Collision Avoidance... RTCA Special Committee 147: Minimum Operational Performance Standards for Traffic Alert and Collision...

75 FR 52590 - Seventy-First Meeting: RTCA Special Committee 147: Minimum Operational Performance Standards for...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-08-26

... Committee 147: Minimum Operational Performance Standards for Traffic Alert and Collision Avoidance Systems... Committee 147: Minimum Operational Performance Standards for Traffic Alert and Collision Avoidance Systems... RTCA Special Committee 147: Minimum Operational Performance Standards for Traffic Alert and Collision...

76 FR 58077 - Seventy-Third Meeting: RTCA Special Committee 147: Minimum Operational Performance Standards for...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-09-19

... Committee 147: Minimum Operational Performance Standards for Traffic Alert and Collision Avoidance Systems... Committee 147 meeting: Minimum Operational Performance Standards for Traffic Alert and Collision Avoidance... RTCA Special Committee 147: Minimum Operational Performance Standards for Traffic Alert and Collision...

12 CFR 1008.113 - Performance standards.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 12 Banks and Banking 8 2013-01-01 2013-01-01 false Performance standards. 1008.113 Section 1008.... Act § 1008.113 Performance standards. (a) For the Bureau to determine that a state is providing effective supervision and enforcement, a supervisory authority must meet the following performance standards...

12 CFR 1008.113 - Performance standards.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 12 Banks and Banking 8 2014-01-01 2014-01-01 false Performance standards. 1008.113 Section 1008.... Act § 1008.113 Performance standards. (a) For the Bureau to determine that a state is providing effective supervision and enforcement, a supervisory authority must meet the following performance standards...

20 CFR 632.89 - Performance standards.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 20 Employees' Benefits 3 2012-04-01 2012-04-01 false Performance standards. 632.89 Section 632.89... EMPLOYMENT AND TRAINING PROGRAMS Program Design and Management § 632.89 Performance standards. The Department of Labor shall establish performance standards for all Native American grantees (section 401(h)(1...

40 CFR 264.343 - Performance standards.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 27 2012-07-01 2012-07-01 false Performance standards. 264.343 Section... Incinerators § 264.343 Performance standards. An incinerator burning hazardous waste must be designed... § 264.345, it will meet the following performance standards: (a)(1) Except as provided in paragraph (a...

12 CFR 1008.113 - Performance standards.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 12 Banks and Banking 8 2012-01-01 2012-01-01 false Performance standards. 1008.113 Section 1008.... Act § 1008.113 Performance standards. (a) For the Bureau to determine that a state is providing effective supervision and enforcement, a supervisory authority must meet the following performance standards...

40 CFR 264.343 - Performance standards.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 25 2010-07-01 2010-07-01 false Performance standards. 264.343 Section... Incinerators § 264.343 Performance standards. An incinerator burning hazardous waste must be designed... § 264.345, it will meet the following performance standards: (a)(1) Except as provided in paragraph (a...

40 CFR 264.343 - Performance standards.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 26 2014-07-01 2014-07-01 false Performance standards. 264.343 Section... Incinerators § 264.343 Performance standards. An incinerator burning hazardous waste must be designed... § 264.345, it will meet the following performance standards: (a)(1) Except as provided in paragraph (a...

40 CFR 264.343 - Performance standards.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 27 2013-07-01 2013-07-01 false Performance standards. 264.343 Section... Incinerators § 264.343 Performance standards. An incinerator burning hazardous waste must be designed... § 264.345, it will meet the following performance standards: (a)(1) Except as provided in paragraph (a...

20 CFR 632.89 - Performance standards.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false Performance standards. 632.89 Section 632.89... EMPLOYMENT AND TRAINING PROGRAMS Program Design and Management § 632.89 Performance standards. The Department of Labor shall establish performance standards for all Native American grantees (section 401(h)(1...

76 FR 7818 - Correction to Notice Soliciting Comments on Proposed Voluntary Product Standard PS 2-10 and...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-02-11

... ``Performance Standard for Wood-Based Structural-Use Panels,'' and appears in the body of the notice. NIST is... correct title of the proposed standard is ``Performance Standard for Wood-Based Structural-Use Panels... Product Standard (PS) 2-04, Performance Standard for Wood-Based Structural-Use Panels. This revised...

76 FR 66886 - Oil and Natural Gas Sector: New Source Performance Standards and National Emission Standards for...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-10-28

...-AP76 Oil and Natural Gas Sector: New Source Performance Standards and National Emission Standards for... and Natural Gas Sector: New Source Performance Standards and National Emission Standards for Hazardous... be charged for copying. World Wide Web. The EPA Web site for this rulemaking is located at: http...

40 CFR 421.194 - Standards of performance for new sources.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 29 2011-07-01 2009-07-01 true Standards of performance for new... Subcategory § 421.194 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Displacement Supernatant. NSPS for the...

40 CFR 421.204 - Standards of performance for new sources.

Code of Federal Regulations, 2010 CFR

2010-07-01

... Subcategory § 421.204 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Spent battery electrolyte. NSPS for the... 40 Protection of Environment 28 2010-07-01 2010-07-01 true Standards of performance for new...

40 CFR 421.254 - Standards of performance for new sources.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 29 2014-07-01 2012-07-01 true Standards of performance for new... Metals and Mercury Subcategory § 421.254 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Smelter wet air...

40 CFR 421.54 - Standards of performance for new sources.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 30 2013-07-01 2012-07-01 true Standards of performance for new... Electrolytic Copper Refining Subcategory § 421.54 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Subpart E...

40 CFR 421.54 - Standards of performance for new sources.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 29 2011-07-01 2009-07-01 true Standards of performance for new... Electrolytic Copper Refining Subcategory § 421.54 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Subpart E...

40 CFR 421.254 - Standards of performance for new sources.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 30 2013-07-01 2012-07-01 true Standards of performance for new... Metals and Mercury Subcategory § 421.254 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Smelter wet air...

40 CFR 421.194 - Standards of performance for new sources.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 29 2014-07-01 2012-07-01 true Standards of performance for new... Subcategory § 421.194 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Displacement Supernatant. NSPS for the...

40 CFR 421.194 - Standards of performance for new sources.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 28 2010-07-01 2010-07-01 true Standards of performance for new... Subcategory § 421.194 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Displacement Supernatant. NSPS for the...

40 CFR 421.254 - Standards of performance for new sources.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 30 2012-07-01 2012-07-01 false Standards of performance for new... Metals and Mercury Subcategory § 421.254 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Smelter wet air...

40 CFR 421.204 - Standards of performance for new sources.

Code of Federal Regulations, 2011 CFR

2011-07-01

... Subcategory § 421.204 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Spent battery electrolyte. NSPS for the... 40 Protection of Environment 29 2011-07-01 2009-07-01 true Standards of performance for new...

40 CFR 421.54 - Standards of performance for new sources.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 28 2010-07-01 2010-07-01 true Standards of performance for new... Electrolytic Copper Refining Subcategory § 421.54 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Subpart E...

40 CFR 421.204 - Standards of performance for new sources.

Code of Federal Regulations, 2014 CFR

2014-07-01

... Subcategory § 421.204 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Spent battery electrolyte. NSPS for the... 40 Protection of Environment 29 2014-07-01 2012-07-01 true Standards of performance for new...

40 CFR 421.204 - Standards of performance for new sources.

Code of Federal Regulations, 2013 CFR

2013-07-01

... Subcategory § 421.204 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Spent battery electrolyte. NSPS for the... 40 Protection of Environment 30 2013-07-01 2012-07-01 true Standards of performance for new...

40 CFR 421.194 - Standards of performance for new sources.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 30 2012-07-01 2012-07-01 false Standards of performance for new... Subcategory § 421.194 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Displacement Supernatant. NSPS for the...

40 CFR 421.54 - Standards of performance for new sources.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 30 2012-07-01 2012-07-01 false Standards of performance for new... Electrolytic Copper Refining Subcategory § 421.54 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Subpart E...

40 CFR 421.194 - Standards of performance for new sources.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 30 2013-07-01 2012-07-01 true Standards of performance for new... Subcategory § 421.194 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Displacement Supernatant. NSPS for the...

40 CFR 421.204 - Standards of performance for new sources.

Code of Federal Regulations, 2012 CFR

2012-07-01

... Subcategory § 421.204 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Spent battery electrolyte. NSPS for the... 40 Protection of Environment 30 2012-07-01 2012-07-01 false Standards of performance for new...

40 CFR 421.54 - Standards of performance for new sources.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 29 2014-07-01 2012-07-01 true Standards of performance for new... Electrolytic Copper Refining Subcategory § 421.54 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Subpart E...

21 CFR 898.12 - Performance standard.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Performance standard. 898.12 Section 898.12 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES PERFORMANCE STANDARD FOR ELECTRODE LEAD WIRES AND PATIENT CABLES § 898.12 Performance standard. (a...

21 CFR 898.12 - Performance standard.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Performance standard. 898.12 Section 898.12 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES PERFORMANCE STANDARD FOR ELECTRODE LEAD WIRES AND PATIENT CABLES § 898.12 Performance standard. (a...

21 CFR 898.12 - Performance standard.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Performance standard. 898.12 Section 898.12 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES PERFORMANCE STANDARD FOR ELECTRODE LEAD WIRES AND PATIENT CABLES § 898.12 Performance standard. (a...

21 CFR 898.12 - Performance standard.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Performance standard. 898.12 Section 898.12 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES PERFORMANCE STANDARD FOR ELECTRODE LEAD WIRES AND PATIENT CABLES § 898.12 Performance standard. (a...

30 CFR 740.19 - Performance standards.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 30 Mineral Resources 3 2012-07-01 2012-07-01 false Performance standards. 740.19 Section 740.19... § 740.19 Performance standards. (a) Operations and reclamation. (1) Surface coal mining and reclamation operations on lands subject to this part shall be conducted in accordance with the performance standards of...

30 CFR 740.19 - Performance standards.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 30 Mineral Resources 3 2011-07-01 2011-07-01 false Performance standards. 740.19 Section 740.19... § 740.19 Performance standards. (a) Operations and reclamation. (1) Surface coal mining and reclamation operations on lands subject to this part shall be conducted in accordance with the performance standards of...

30 CFR 750.16 - Performance standards.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 30 Mineral Resources 3 2014-07-01 2014-07-01 false Performance standards. 750.16 Section 750.16... Performance standards. After OSM issues a permit under this part, a person conducting surface coal mining... shall adhere to the performance standards of 30 CFR chapter VII, subchapter B. [49 FR 38477, Sept. 28...

30 CFR 750.16 - Performance standards.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 30 Mineral Resources 3 2013-07-01 2013-07-01 false Performance standards. 750.16 Section 750.16... Performance standards. After OSM issues a permit under this part, a person conducting surface coal mining... shall adhere to the performance standards of 30 CFR chapter VII, subchapter B. [49 FR 38477, Sept. 28...

30 CFR 750.16 - Performance standards.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 30 Mineral Resources 3 2011-07-01 2011-07-01 false Performance standards. 750.16 Section 750.16... Performance standards. After OSM issues a permit under this part, a person conducting surface coal mining... shall adhere to the performance standards of 30 CFR chapter VII, subchapter B. [49 FR 38477, Sept. 28...

30 CFR 740.19 - Performance standards.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 30 Mineral Resources 3 2014-07-01 2014-07-01 false Performance standards. 740.19 Section 740.19... § 740.19 Performance standards. (a) Operations and reclamation. (1) Surface coal mining and reclamation operations on lands subject to this part shall be conducted in accordance with the performance standards of...

21 CFR 898.12 - Performance standard.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Performance standard. 898.12 Section 898.12 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES PERFORMANCE STANDARD FOR ELECTRODE LEAD WIRES AND PATIENT CABLES § 898.12 Performance standard. (a...

30 CFR 740.19 - Performance standards.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 30 Mineral Resources 3 2013-07-01 2013-07-01 false Performance standards. 740.19 Section 740.19... § 740.19 Performance standards. (a) Operations and reclamation. (1) Surface coal mining and reclamation operations on lands subject to this part shall be conducted in accordance with the performance standards of...

30 CFR 750.16 - Performance standards.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 30 Mineral Resources 3 2012-07-01 2012-07-01 false Performance standards. 750.16 Section 750.16... Performance standards. After OSM issues a permit under this part, a person conducting surface coal mining... shall adhere to the performance standards of 30 CFR chapter VII, subchapter B. [49 FR 38477, Sept. 28...

30 CFR 750.16 - Performance standards.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 30 Mineral Resources 3 2010-07-01 2010-07-01 false Performance standards. 750.16 Section 750.16... Performance standards. After OSM issues a permit under this part, a person conducting surface coal mining... shall adhere to the performance standards of 30 CFR chapter VII, subchapter B. [49 FR 38477, Sept. 28...

30 CFR 740.19 - Performance standards.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 30 Mineral Resources 3 2010-07-01 2010-07-01 false Performance standards. 740.19 Section 740.19... § 740.19 Performance standards. (a) Operations and reclamation. (1) Surface coal mining and reclamation operations on lands subject to this part shall be conducted in accordance with the performance standards of...

40 CFR 60.372 - Standards for lead.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 7 2013-07-01 2013-07-01 false Standards for lead. 60.372 Section 60...) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Lead-Acid Battery Manufacturing Plants § 60.372 Standards for lead. (a) On and after the date on which the performance test...

40 CFR 60.372 - Standards for lead.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 7 2012-07-01 2012-07-01 false Standards for lead. 60.372 Section 60...) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Lead-Acid Battery Manufacturing Plants § 60.372 Standards for lead. (a) On and after the date on which the performance test...

40 CFR 60.372 - Standards for lead.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 7 2014-07-01 2014-07-01 false Standards for lead. 60.372 Section 60...) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Lead-Acid Battery Manufacturing Plants § 60.372 Standards for lead. (a) On and after the date on which the performance test...

40 CFR 60.372 - Standards for lead.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 6 2011-07-01 2011-07-01 false Standards for lead. 60.372 Section 60...) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Lead-Acid Battery Manufacturing Plants § 60.372 Standards for lead. (a) On and after the date on which the performance test...

40 CFR 60.474 - Test methods and procedures.

Code of Federal Regulations, 2010 CFR

2010-07-01

... compliance with the mass standard for the blowing still. If the Administrator finds that the facility was in compliance with the mass standard during the performance test but failed to meet the zero opacity standard... (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Asphalt...

40 CFR 60.474 - Test methods and procedures.

Code of Federal Regulations, 2011 CFR

2011-07-01

... compliance with the mass standard for the blowing still. If the Administrator finds that the facility was in compliance with the mass standard during the performance test but failed to meet the zero opacity standard... (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Asphalt...

40 CFR 421.84 - Standards of performance for new sources.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 29 2014-07-01 2012-07-01 true Standards of performance for new... Subcategory § 421.84 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Subpart H—Zinc Reduction Furnace Wet Air...

40 CFR 421.84 - Standards of performance for new sources.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 30 2013-07-01 2012-07-01 true Standards of performance for new... Subcategory § 421.84 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Subpart H—Zinc Reduction Furnace Wet Air...

40 CFR 421.84 - Standards of performance for new sources.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 30 2012-07-01 2012-07-01 false Standards of performance for new... Subcategory § 421.84 Standards of performance for new sources. Any new source subject to this subpart shall achieve the following new source performance standards: (a) Subpart H—Zinc Reduction Furnace Wet Air...

20 CFR 1001.124 - Standards of performance governing State agency cooperation and coordination with other agencies...

Code of Federal Regulations, 2010 CFR

2010-04-01

... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false Standards of performance governing State..., DEPARTMENT OF LABOR SERVICES FOR VETERANS Standards of Performance Governing State Agency Services to Veterans and Eligible Persons § 1001.124 Standards of performance governing State agency cooperation and...

Mathematics Content Standards Benchmarks and Performance Standards

ERIC Educational Resources Information Center

New Mexico Public Education Department, 2008

2008-01-01

New Mexico Mathematics Content Standards, Benchmarks, and Performance Standards identify what students should know and be able to do across all grade levels, forming a spiraling framework in the sense that many skills, once introduced, develop over time. While the Performance Standards are set forth at grade-specific levels, they do not exist as…

20 CFR 404.1643 - Performance accuracy standard.

Code of Federal Regulations, 2011 CFR

2011-04-01

... DISABILITY INSURANCE (1950- ) Determinations of Disability Performance Standards § 404.1643 Performance... well as the correctness of the decision. For example, if a particular item of medical evidence should... case, that is a performance error. Performance accuracy, therefore, is a higher standard than...

Intelligence Level Performance Standards Research for Autonomous Vehicles

PubMed Central

Bostelman, Roger B.; Hong, Tsai H.; Messina, Elena

2017-01-01

United States and European safety standards have evolved to protect workers near Automatic Guided Vehicles (AGV’s). However, performance standards for AGV’s and mobile robots have only recently begun development. Lessons can be learned from research and standards efforts for mobile robots applied to emergency response and military applications. Research challenges, tests and evaluations, and programs to develop higher intelligence levels for vehicles can also used to guide industrial AGV developments towards more adaptable and intelligent systems. These other efforts also provide useful standards development criteria for AGV performance test methods. Current standards areas being considered for AGVs are for docking, navigation, obstacle avoidance, and the ground truth systems that measure performance. This paper provides a look to the future with standards developments in both the performance of vehicles and the dynamic perception systems that measure intelligent vehicle performance. PMID:28649189

Intelligence Level Performance Standards Research for Autonomous Vehicles.

PubMed

Bostelman, Roger B; Hong, Tsai H; Messina, Elena

2015-01-01

United States and European safety standards have evolved to protect workers near Automatic Guided Vehicles (AGV's). However, performance standards for AGV's and mobile robots have only recently begun development. Lessons can be learned from research and standards efforts for mobile robots applied to emergency response and military applications. Research challenges, tests and evaluations, and programs to develop higher intelligence levels for vehicles can also used to guide industrial AGV developments towards more adaptable and intelligent systems. These other efforts also provide useful standards development criteria for AGV performance test methods. Current standards areas being considered for AGVs are for docking, navigation, obstacle avoidance, and the ground truth systems that measure performance. This paper provides a look to the future with standards developments in both the performance of vehicles and the dynamic perception systems that measure intelligent vehicle performance.

Setting Performance Standards for Technical and Nontechnical Competence in General Surgery.

PubMed

Szasz, Peter; Bonrath, Esther M; Louridas, Marisa; Fecso, Andras B; Howe, Brett; Fehr, Adam; Ott, Michael; Mack, Lloyd A; Harris, Kenneth A; Grantcharov, Teodor P

2017-07-01

The objectives of this study were to (1) create a technical and nontechnical performance standard for the laparoscopic cholecystectomy, (2) assess the classification accuracy and (3) credibility of these standards, (4) determine a trainees' ability to meet both standards concurrently, and (5) delineate factors that predict standard acquisition. Scores on performance assessments are difficult to interpret in the absence of established standards. Trained raters observed General Surgery residents performing laparoscopic cholecystectomies using the Objective Structured Assessment of Technical Skill (OSATS) and the Objective Structured Assessment of Non-Technical Skills (OSANTS) instruments, while as also providing a global competent/noncompetent decision for each performance. The global decision was used to divide the trainees into 2 contrasting groups and the OSATS or OSANTS scores were graphed per group to determine the performance standard. Parametric statistics were used to determine classification accuracy and concurrent standard acquisition, receiver operator characteristic (ROC) curves were used to delineate predictive factors. Thirty-six trainees were observed 101 times. The technical standard was an OSATS of 21.04/35.00 and the nontechnical standard an OSANTS of 22.49/35.00. Applying these standards, competent/noncompetent trainees could be discriminated in 94% of technical and 95% of nontechnical performances (P < 0.001). A 21% discordance between technically and nontechnically competent trainees was identified (P < 0.001). ROC analysis demonstrated case experience and trainee level were both able to predict achieving the standards with an area under the curve (AUC) between 0.83 and 0.96 (P < 0.001). The present study presents defensible standards for technical and nontechnical performance. Such standards are imperative to implementing summative assessments into surgical training.

National Unmanned Aerial System Standardized Performance Testing and Rating (NUSTAR)

NASA Technical Reports Server (NTRS)

Kopardekar, Parimal

2016-01-01

The overall objective of the NUSTAR Capability is to offer standardized tests and scenario conditions to assess performance of the UAS. The following are goals of the NU-STAR: 1. Create a prototype standardized tests and scenarios that vehicles can be tested against. 2. Identify key performance parameters of all UAS and their standardized measurement strategy. 3. Develop standardized performance reporting method (e.g., consumer report style) to assist prospective buyers. 4. Identify key performance metrics that could be used by judged towards overall safety of the UAS and operations. 5. If vehicle certification standard is made by a regulatory agency, the performance of individual UAS could be compared against the minimum requirement (e.g., sense and avoid detection time, stopping distance, kinetic energy, etc.).

21 CFR 861.7 - Contents of standards.

Code of Federal Regulations, 2011 CFR

2011-04-01

... DEVICES PROCEDURES FOR PERFORMANCE STANDARDS DEVELOPMENT General § 861.7 Contents of standards. Any performance standard established under this part will include such provisions as the Food and Drug... address (but need not be limited to): (a) Performance characteristics of the device; (b) The design...

46 CFR 197.520 - Performance standard.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 46 Shipping 7 2014-10-01 2014-10-01 false Performance standard. 197.520 Section 197.520 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) MARINE OCCUPATIONAL SAFETY AND HEALTH STANDARDS GENERAL PROVISIONS Benzene § 197.520 Performance standard. No person may be subjected to a personal...

46 CFR 197.520 - Performance standard.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 46 Shipping 7 2013-10-01 2013-10-01 false Performance standard. 197.520 Section 197.520 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) MARINE OCCUPATIONAL SAFETY AND HEALTH STANDARDS GENERAL PROVISIONS Benzene § 197.520 Performance standard. No person may be subjected to a personal...

46 CFR 197.520 - Performance standard.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 46 Shipping 7 2010-10-01 2010-10-01 false Performance standard. 197.520 Section 197.520 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) MARINE OCCUPATIONAL SAFETY AND HEALTH STANDARDS GENERAL PROVISIONS Benzene § 197.520 Performance standard. No person may be subjected to a personal...

46 CFR 197.520 - Performance standard.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 46 Shipping 7 2011-10-01 2011-10-01 false Performance standard. 197.520 Section 197.520 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) MARINE OCCUPATIONAL SAFETY AND HEALTH STANDARDS GENERAL PROVISIONS Benzene § 197.520 Performance standard. No person may be subjected to a personal...

46 CFR 197.520 - Performance standard.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 46 Shipping 7 2012-10-01 2012-10-01 false Performance standard. 197.520 Section 197.520 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) MARINE OCCUPATIONAL SAFETY AND HEALTH STANDARDS GENERAL PROVISIONS Benzene § 197.520 Performance standard. No person may be subjected to a personal...

40 CFR 60.474 - Test methods and procedures.

Code of Federal Regulations, 2014 CFR

2014-07-01

... a finding concerning compliance with the mass standard for the blowing still. If the Administrator finds that the facility was in compliance with the mass standard during the performance test but failed... (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Asphalt...

40 CFR 60.474 - Test methods and procedures.

Code of Federal Regulations, 2013 CFR

2013-07-01

... a finding concerning compliance with the mass standard for the blowing still. If the Administrator finds that the facility was in compliance with the mass standard during the performance test but failed... (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Asphalt...

40 CFR 463.34 - New source performance standards.

Code of Federal Regulations, 2012 CFR

2012-07-01

... performance standards (i.e., mass of pollutant discharged), which are calculated by multiplying the average... 40 Protection of Environment 31 2012-07-01 2012-07-01 false New source performance standards. 463... GUIDELINES AND STANDARDS (CONTINUED) PLASTICS MOLDING AND FORMING POINT SOURCE CATEGORY Finishing Water...

40 CFR 60.474 - Test methods and procedures.

Code of Federal Regulations, 2012 CFR

2012-07-01

... a finding concerning compliance with the mass standard for the blowing still. If the Administrator finds that the facility was in compliance with the mass standard during the performance test but failed... (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Asphalt...

40 CFR 463.24 - New source performance standards.

Code of Federal Regulations, 2012 CFR

2012-07-01

... performance standards (i.e., mass of pollutant discharged) calculated by multiplying the average process water... 40 Protection of Environment 31 2012-07-01 2012-07-01 false New source performance standards. 463... GUIDELINES AND STANDARDS (CONTINUED) PLASTICS MOLDING AND FORMING POINT SOURCE CATEGORY Cleaning Water...

A National Initiative to Advance School Mental Health Performance Measurement in the US

ERIC Educational Resources Information Center

Connors, Elizabeth Halsted; Stephan, Sharon Hoover; Lever, Nancy; Ereshefsky, Sabrina; Mosby, Amanda; Bohnenkamp, Jill

2016-01-01

Standardized health performance measurement has increasingly become an imperative for assuring quality standards in national health care systems. As compared to somatic health performance measures, behavioral health performance measures are less developed. There currently is no national standardized performance measurement system for monitoring…

40 CFR 420.44 - New source performance standards (NSPS).

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 28 2010-07-01 2010-07-01 true New source performance standards (NSPS... GUIDELINES AND STANDARDS IRON AND STEEL MANUFACTURING POINT SOURCE CATEGORY Steelmaking Subcategory § 420.44 New source performance standards (NSPS). The discharge of wastewater pollutants from any new source...

10 CFR 433.4 - Energy efficiency performance standard.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 10 Energy 3 2014-01-01 2014-01-01 false Energy efficiency performance standard. 433.4 Section 433.4 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY STANDARDS FOR NEW FEDERAL COMMERCIAL AND MULTI-FAMILY HIGH-RISE RESIDENTIAL BUILDINGS § 433.4 Energy efficiency performance standard...

10 CFR 433.4 - Energy efficiency performance standard.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 10 Energy 3 2012-01-01 2012-01-01 false Energy efficiency performance standard. 433.4 Section 433.4 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY STANDARDS FOR NEW FEDERAL COMMERCIAL AND MULTI-FAMILY HIGH-RISE RESIDENTIAL BUILDINGS § 433.4 Energy efficiency performance standard...

10 CFR 433.4 - Energy efficiency performance standard.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 10 Energy 3 2013-01-01 2013-01-01 false Energy efficiency performance standard. 433.4 Section 433.4 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY EFFICIENCY STANDARDS FOR NEW FEDERAL COMMERCIAL AND MULTI-FAMILY HIGH-RISE RESIDENTIAL BUILDINGS § 433.4 Energy efficiency performance standard...

40 CFR 455.25 - New source performance standards (NSPS).

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 31 2012-07-01 2012-07-01 false New source performance standards (NSPS... GUIDELINES AND STANDARDS (CONTINUED) PESTICIDE CHEMICALS Organic Pesticide Chemicals Manufacturing Subcategory § 455.25 New source performance standards (NSPS). (a) Any new source subject to this subpart which...

40 CFR 463.14 - New source performance standards.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 31 2012-07-01 2012-07-01 false New source performance standards. 463... GUIDELINES AND STANDARDS (CONTINUED) PLASTICS MOLDING AND FORMING POINT SOURCE CATEGORY Contact Cooling and Heating Water Subcategory § 463.14 New source performance standards. (a) NSPS for bis(2-ethylhexyl...

40 CFR 455.25 - New source performance standards (NSPS).

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 29 2010-07-01 2010-07-01 false New source performance standards (NSPS... GUIDELINES AND STANDARDS PESTICIDE CHEMICALS Organic Pesticide Chemicals Manufacturing Subcategory § 455.25 New source performance standards (NSPS). (a) Any new source subject to this subpart which discharges...

Hearing the voices of alternatively certified teachers in Texas: Narratives of teaching English language learners in urban secondary mainstream classrooms

NASA Astrophysics Data System (ADS)

Zannou, Yetunde Mobola

In Texas, nearly half of all new teachers are alternatively certified (AC) whilst English language learners (ELL) are over one-third of the public school population in some districts. As this trend continues, the likelihood that AC teachers will teach ELLs increases and alters what Texas teachers must know upon entering the classroom. This research explores teacher knowledge and beliefs about teaching ELLs through constructivist and narrative lenses. Four AC science teachers in two diverse school districts participated in in-depth interviews and reflective interviews following classroom observations to answer the research questions: (1) how do AC teachers describe and interpret their acts of teaching ELLs in mainstream classrooms; and (2) how do AC teachers describe and interpret their learning to teach ELLs in mainstream classrooms. Data were transcribed and analyzed using thematic narrative methods. This study found that participants saw ELL instruction as: (1) "just good teaching" strategies, (2) consisting primarily of cultural awareness and consideration for student comfort, and (3) less necessary in science where all students must learn the language. The most experienced teacher was the only participant to reference specific linguistic knowledge in describing ELL instruction. Many of the teachers described their work with ELL students as giving them an opportunity to improve their lives, which was consistent with their overall teaching philosophy and reason for entering the profession. Participant narratives about learning to teach ELLs described personal experience and person-to-person discussions as primary resources of knowledge. District support was generally described as unhelpful or incomplete. Participants portrayed their AC program as helpful in preparing them to work with ELL students, but everyone desired more relevant information from the program and more grade-appropriate strategies from the district. Participant narratives reveal AC teachers needed a pragmatic and less theoretical understanding of diversity during pre-service training. Participant tendency to draw upon "common sense", affective, and practical strategies in teaching ELL students in lieu of the state-mandated English language proficiency standards (ELPS) suggests AC programs should have teachers articulate and discuss their beliefs about ELL instruction in order to provide training targeted towards misconceptions about language development, particularly in science.

High-throughput purification of recombinant proteins using self-cleaving intein tags.

PubMed

Coolbaugh, M J; Shakalli Tang, M J; Wood, D W

2017-01-01

High throughput methods for recombinant protein production using E. coli typically involve the use of affinity tags for simple purification of the protein of interest. One drawback of these techniques is the occasional need for tag removal before study, which can be hard to predict. In this work, we demonstrate two high throughput purification methods for untagged protein targets based on simple and cost-effective self-cleaving intein tags. Two model proteins, E. coli beta-galactosidase (βGal) and superfolder green fluorescent protein (sfGFP), were purified using self-cleaving versions of the conventional chitin-binding domain (CBD) affinity tag and the nonchromatographic elastin-like-polypeptide (ELP) precipitation tag in a 96-well filter plate format. Initial tests with shake flask cultures confirmed that the intein purification scheme could be scaled down, with >90% pure product generated in a single step using both methods. The scheme was then validated in a high throughput expression platform using 24-well plate cultures followed by purification in 96-well plates. For both tags and with both target proteins, the purified product was consistently obtained in a single-step, with low well-to-well and plate-to-plate variability. This simple method thus allows the reproducible production of highly pure untagged recombinant proteins in a convenient microtiter plate format. Copyright © 2016 Elsevier Inc. All rights reserved.

Physical Interactions and Expression Quantitative Traits Loci Identify Regulatory Connections for Obesity and Type 2 Diabetes Associated SNPs

PubMed Central

Fadason, Tayaza; Ekblad, Cameron; Ingram, John R.; Schierding, William S.; O'Sullivan, Justin M.

2017-01-01

The mechanisms that underlie the association between obesity and type 2 diabetes are not fully understood. Here, we investigated the role of the 3D genome organization in the pathogeneses of obesity and type-2 diabetes. We interpreted the combined and differential impacts of 196 diabetes and 390 obesity associated single nucleotide polymorphisms (SNPs) by integrating data on the genes with which they physically interact (as captured by Hi-C) and the functional [i.e., expression quantitative trait loci (eQTL)] outcomes associated with these interactions. We identified 861 spatially regulated genes (e.g., AP3S2, ELP5, SVIP, IRS1, FADS2, WFS1, RBM6, HORMAD1, PYROXD2), which are enriched in tissues (e.g., adipose, skeletal muscle, pancreas) and biological processes and canonical pathways (e.g., lipid metabolism, leptin, and glucose-insulin signaling pathways) that are important for the pathogenesis of type 2 diabetes and obesity. Our discovery-based approach also identifies enrichment for eQTL SNP-gene interactions in tissues that are not classically associated with diabetes or obesity. We propose that the combinatorial action of active obesity and diabetes spatial eQTL SNPs on their gene pairs within different tissues reduces the ability of these tissues to contribute to the maintenance of a healthy energy metabolism. PMID:29081791

Investigating the interplay between substrate stiffness and ligand chemistry in directing mesenchymal stem cell differentiation within 3D macro-porous substrates.

PubMed

Haugh, Matthew G; Vaughan, Ted J; Madl, Christopher M; Raftery, Rosanne M; McNamara, Laoise M; O'Brien, Fergal J; Heilshorn, Sarah C

2018-07-01

Dimensionality can have a profound impact on stiffness-mediated differentiation of mesenchymal stem cells (MSCs). However, while we have begun to understand cellular response when encapsulated within 3D substrates, the behavior of cells within macro-porous substrates is relatively underexplored. The goal of this study was to determine the influence of macro-porous topographies on stiffness-mediated differentiation of MSCs. We developed macro-porous recombinant elastin-like protein (ELP) substrates that allow independent control of mechanical properties and ligand chemistry. We then used computational modeling to probe the impact of pore topography on the mechanical stimulus that cells are exposed to within these substrates, and finally we investigated stiffness induced biases towards adipogenic and osteogenic differentiation of MSCs within macro-porous substrates. Computational modeling revealed that there is significant heterogeneity in the mechanical stimuli that cells are exposed to within porous substrates and that this heterogeneity is predominantly due to the wide range of possible cellular orientations within the pores. Surprisingly, MSCs grown within 3D porous substrates respond to increasing substrate stiffness by up-regulating both osteogenesis and adipogenesis. These results demonstrate that within porous substrates the behavior of MSCs diverges from previously observed responses to substrate stiffness, emphasizing the importance of topography as a determinant of cellular behavior. Copyright © 2018 Elsevier Ltd. All rights reserved.

Light pollution reduces activity, food consumption and growth rates in a sandy beach invertebrate.

PubMed

Luarte, T; Bonta, C C; Silva-Rodriguez, E A; Quijón, P A; Miranda, C; Farias, A A; Duarte, C

2016-11-01

The continued growth of human activity and infrastructure has translated into a widespread increase in light pollution. Natural daylight and moonlight cycles play a fundamental role for many organisms and ecological processes, so an increase in light pollution may have profound effects on communities and ecosystem services. Studies assessing ecological light pollution (ELP) effects on sandy beach organisms have lagged behind the study of other sources of disturbance. Hence, we assessed the influence of this stressor on locomotor activity, foraging behavior, absorption efficiency and growth rate of adults of the talitrid amphipod Orchestoidea tuberculata. In the field, an artificial light system was assembled to assess the local influence of artificial light conditions on the amphipod's locomotor activity and use of food patches in comparison to natural (ambient) conditions. Meanwhile in the laboratory, two experimental chambers were set to assess amphipod locomotor activity, consumption rates, absorption efficiency and growth under artificial light in comparison to natural light-dark cycles. Our results indicate that artificial light have significantly adverse effects on the activity patterns and foraging behavior of the amphipods, resulting on reduced consumption and growth rates. Given the steady increase in artificial light pollution here and elsewhere, sandy beach communities could be negatively affected, with unexpected consequences for the whole ecosystem. Copyright © 2016 Elsevier Ltd. All rights reserved.

rRNA fragmentation induced by a yeast killer toxin.

PubMed

Kast, Alene; Klassen, Roland; Meinhardt, Friedhelm

2014-02-01

Virus like dsDNA elements (VLE) in yeast were previously shown to encode the killer toxins PaT and zymocin, which target distinct tRNA species via specific anticodon nuclease (ACNase) activities. Here, we characterize a third member of the VLE-encoded toxins, PiT from Pichia inositovora, and identify PiOrf4 as the cytotoxic subunit by conditional expression in Saccharomyces cerevisiae. In contrast to the tRNA targeting toxins, however, neither a change of the wobble uridine modification status by introduction of elp3 or trm9 mutations nor tRNA overexpression rescued from PiOrf4 toxicity. Consistent with a distinct RNA target, expression of PiOrf4 causes specific fragmentation of the 25S and 18S rRNA. A stable cleavage product comprising the first ∼ 130 nucleotides of the 18S rRNA was purified and characterized by linker ligation and subsequent reverse transcription; 3'-termini were mapped to nucleotide 131 and 132 of the 18S rRNA sequence, a region showing some similarity to the anticodon loop of tRNA(Glu)(UUC), the zymocin target. PiOrf4 residues Glu9 and His214, corresponding to catalytic sites Glu9 and His209 in the ACNase subunit of zymocin are essential for in vivo toxicity and rRNA fragmentation, raising the possibility of functionally conserved RNase modules in both proteins. © 2013 John Wiley & Sons Ltd.

TLR-activated repression of Fe-S cluster biogenesis drives a metabolic shift and alters histone and tubulin acetylation.

PubMed

Tong, Wing-Hang; Maio, Nunziata; Zhang, De-Liang; Palmieri, Erika M; Ollivierre, Hayden; Ghosh, Manik C; McVicar, Daniel W; Rouault, Tracey A

2018-05-22

Given the essential roles of iron-sulfur (Fe-S) cofactors in mediating electron transfer in the mitochondrial respiratory chain and supporting heme biosynthesis, mitochondrial dysfunction is a common feature in a growing list of human Fe-S cluster biogenesis disorders, including Friedreich ataxia and GLRX5-related sideroblastic anemia. Here, our studies showed that restriction of Fe-S cluster biogenesis not only compromised mitochondrial oxidative metabolism but also resulted in decreased overall histone acetylation and increased H3K9me3 levels in the nucleus and increased acetylation of α-tubulin in the cytosol by decreasing the lipoylation of the pyruvate dehydrogenase complex, decreasing levels of succinate dehydrogenase and the histone acetyltransferase ELP3, and increasing levels of the tubulin acetyltransferase MEC17. Previous studies have shown that the metabolic shift in Toll-like receptor (TLR)-activated myeloid cells involves rapid activation of glycolysis and subsequent mitochondrial respiratory failure due to nitric oxide (NO)-mediated damage to Fe-S proteins. Our studies indicated that TLR activation also actively suppresses many components of the Fe-S cluster biogenesis machinery, which exacerbates NO-mediated damage to Fe-S proteins by interfering with cluster recovery. These results reveal new regulatory pathways and novel roles of the Fe-S cluster biogenesis machinery in modifying the epigenome and acetylome and provide new insights into the etiology of Fe-S cluster biogenesis disorders.

77 FR 73968 - Reconsideration of Certain New Source and Startup/Shutdown Issues: National Emission Standards...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-12-12

... Units and Standards of Performance for Fossil-Fuel-Fired Electric Utility, Industrial-Commercial...- and Oil-fired Electric Utility Steam Generating Units and Standards of Performance for Fossil-Fuel... Units and Standards of Performance for Fossil-Fuel-Fired Electric Utility, Industrial-Commercial...

40 CFR 1700.14 - Marine Pollution Control Device (MPCD) Performance Standards. [Reserved

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 34 2012-07-01 2012-07-01 false Marine Pollution Control Device (MPCD... UNIFORM NATIONAL DISCHARGE STANDARDS FOR VESSELS OF THE ARMED FORCES Marine Pollution Control Device (MPCD) Performance Standards § 1700.14 Marine Pollution Control Device (MPCD) Performance Standards. [Reserved] ...

40 CFR 1700.14 - Marine Pollution Control Device (MPCD) Performance Standards. [Reserved

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 34 2013-07-01 2013-07-01 false Marine Pollution Control Device (MPCD... UNIFORM NATIONAL DISCHARGE STANDARDS FOR VESSELS OF THE ARMED FORCES Marine Pollution Control Device (MPCD) Performance Standards § 1700.14 Marine Pollution Control Device (MPCD) Performance Standards. [Reserved] ...

40 CFR 1700.14 - Marine Pollution Control Device (MPCD) Performance Standards. [Reserved

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 33 2014-07-01 2014-07-01 false Marine Pollution Control Device (MPCD... UNIFORM NATIONAL DISCHARGE STANDARDS FOR VESSELS OF THE ARMED FORCES Marine Pollution Control Device (MPCD) Performance Standards § 1700.14 Marine Pollution Control Device (MPCD) Performance Standards. [Reserved] ...

40 CFR 60.184 - Standard for visible emissions.

Code of Federal Regulations, 2010 CFR

2010-07-01

... (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Primary Lead Smelters § 60.184 Standard for visible emissions. (a) On and after the date on which the performance test... furnace, or sintering machine discharge end any visible emissions which exhibit greater than 20 percent...

40 CFR 60.722 - Standards for volatile organic compounds.

Code of Federal Regulations, 2010 CFR

2010-07-01

... PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Industrial Surface Coating: Surface Coating of Plastic Parts for Business Machines § 60.722 Standards for... the date on which the initial performance test, required by §§ 60.8 and 60.723 is completed, but not...

40 CFR 60.722 - Standards for volatile organic compounds.

Code of Federal Regulations, 2011 CFR

2011-07-01

... PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Industrial Surface Coating: Surface Coating of Plastic Parts for Business Machines § 60.722 Standards for... the date on which the initial performance test, required by §§ 60.8 and 60.723 is completed, but not...

40 CFR 60.184 - Standard for visible emissions.

Code of Federal Regulations, 2011 CFR

2011-07-01

... (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Primary Lead Smelters § 60.184 Standard for visible emissions. (a) On and after the date on which the performance test... furnace, or sintering machine discharge end any visible emissions which exhibit greater than 20 percent...

40 CFR 1700.14 - Marine Pollution Control Device (MPCD) Performance Standards. [Reserved

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 33 2011-07-01 2011-07-01 false Marine Pollution Control Device (MPCD... UNIFORM NATIONAL DISCHARGE STANDARDS FOR VESSELS OF THE ARMED FORCES Marine Pollution Control Device (MPCD) Performance Standards § 1700.14 Marine Pollution Control Device (MPCD) Performance Standards. [Reserved] ...

40 CFR 1700.14 - Marine Pollution Control Device (MPCD) Performance Standards. [Reserved

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 32 2010-07-01 2010-07-01 false Marine Pollution Control Device (MPCD... UNIFORM NATIONAL DISCHARGE STANDARDS FOR VESSELS OF THE ARMED FORCES Marine Pollution Control Device (MPCD) Performance Standards § 1700.14 Marine Pollution Control Device (MPCD) Performance Standards. [Reserved] ...

40 CFR 464.34 - New source performance standards.

Code of Federal Regulations, 2012 CFR

2012-07-01

...-continuous dischargers, annual average mass standards and maximum day and maximum for monthly average concentration (mg/l) standards shall apply. Concentration standards and annual average mass standards shall only... 40 Protection of Environment 31 2012-07-01 2012-07-01 false New source performance standards. 464...

40 CFR 464.34 - New source performance standards.

Code of Federal Regulations, 2014 CFR

2014-07-01

...-continuous dischargers, annual average mass standards and maximum day and maximum for monthly average concentration (mg/l) standards shall apply. Concentration standards and annual average mass standards shall only... 40 Protection of Environment 30 2014-07-01 2014-07-01 false New source performance standards. 464...

40 CFR 464.34 - New source performance standards.

Code of Federal Regulations, 2013 CFR

2013-07-01

...-continuous dischargers, annual average mass standards and maximum day and maximum for monthly average concentration (mg/l) standards shall apply. Concentration standards and annual average mass standards shall only... 40 Protection of Environment 31 2013-07-01 2013-07-01 false New source performance standards. 464...

40 CFR 52.939 - Original identification of plan section.

Code of Federal Regulations, 2012 CFR

2012-07-01

... Performance for Existing Solvent Metal Cleaning Equipment. (7) Regulation 6.19 Standard of Performance for Existing Metal Furniture Surface Coating Operations. (8) Regulation 6.23 Standard of Performance for...) Regulation 6.31 Standard of Performance for Existing Miscellaneous Metal Parts and Products Surface Coating...

40 CFR 52.939 - Original identification of plan section.

Code of Federal Regulations, 2013 CFR

2013-07-01

... Performance for Existing Solvent Metal Cleaning Equipment. (7) Regulation 6.19 Standard of Performance for Existing Metal Furniture Surface Coating Operations. (8) Regulation 6.23 Standard of Performance for...) Regulation 6.31 Standard of Performance for Existing Miscellaneous Metal Parts and Products Surface Coating...

40 CFR 52.939 - Original identification of plan section.

Code of Federal Regulations, 2011 CFR

2011-07-01

... Performance for Existing Solvent Metal Cleaning Equipment. (7) Regulation 6.19 Standard of Performance for Existing Metal Furniture Surface Coating Operations. (8) Regulation 6.23 Standard of Performance for...) Regulation 6.31 Standard of Performance for Existing Miscellaneous Metal Parts and Products Surface Coating...

33 CFR 332.5 - Ecological performance standards.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Ecological performance standards..., DEPARTMENT OF DEFENSE COMPENSATORY MITIGATION FOR LOSSES OF AQUATIC RESOURCES § 332.5 Ecological performance... objective and verifiable. Ecological performance standards must be based on the best available science that...

40 CFR 426.15 - Standards of performance for new sources.

Code of Federal Regulations, 2013 CFR

2013-07-01

... Fiberglass Subcategory § 426.15 Standards of performance for new sources. The following standards of performance establish the quantity or quality of pollutants or pollutant properties which may be discharged by...

40 CFR 426.15 - Standards of performance for new sources.

Code of Federal Regulations, 2012 CFR

2012-07-01

... Fiberglass Subcategory § 426.15 Standards of performance for new sources. The following standards of performance establish the quantity or quality of pollutants or pollutant properties which may be discharged by...

40 CFR 426.15 - Standards of performance for new sources.

Code of Federal Regulations, 2014 CFR

2014-07-01

... Fiberglass Subcategory § 426.15 Standards of performance for new sources. The following standards of performance establish the quantity or quality of pollutants or pollutant properties which may be discharged by...

40 CFR 443.15 - Standards of performance for new sources.

Code of Federal Regulations, 2010 CFR

2010-07-01

... PERFORMANCE AND PRETREATMENT STANDARDS FOR NEW SOURCES FOR THE PAVING AND ROOFING MATERIALS (TARS AND ASPHALT) POINT SOURCE CATEGORY Asphalt Emulsion Subcategory § 443.15 Standards of performance for new sources...

40 CFR 443.25 - Standards of performance for new sources.

Code of Federal Regulations, 2011 CFR

2011-07-01

... PERFORMANCE AND PRETREATMENT STANDARDS FOR NEW SOURCES FOR THE PAVING AND ROOFING MATERIALS (TARS AND ASPHALT) POINT SOURCE CATEGORY Asphalt Concrete Subcategory § 443.25 Standards of performance for new sources...

40 CFR 443.45 - Standards of performance for new sources.

Code of Federal Regulations, 2010 CFR

2010-07-01

... PERFORMANCE AND PRETREATMENT STANDARDS FOR NEW SOURCES FOR THE PAVING AND ROOFING MATERIALS (TARS AND ASPHALT) POINT SOURCE CATEGORY Linoleum and Printed Asphalt Felt Subcategory § 443.45 Standards of performance...

40 CFR 443.25 - Standards of performance for new sources.

Code of Federal Regulations, 2010 CFR

2010-07-01

... PERFORMANCE AND PRETREATMENT STANDARDS FOR NEW SOURCES FOR THE PAVING AND ROOFING MATERIALS (TARS AND ASPHALT) POINT SOURCE CATEGORY Asphalt Concrete Subcategory § 443.25 Standards of performance for new sources...

40 CFR 443.45 - Standards of performance for new sources.

Code of Federal Regulations, 2011 CFR

2011-07-01

... PERFORMANCE AND PRETREATMENT STANDARDS FOR NEW SOURCES FOR THE PAVING AND ROOFING MATERIALS (TARS AND ASPHALT) POINT SOURCE CATEGORY Linoleum and Printed Asphalt Felt Subcategory § 443.45 Standards of performance...

40 CFR 443.15 - Standards of performance for new sources.

Code of Federal Regulations, 2011 CFR

2011-07-01

... PERFORMANCE AND PRETREATMENT STANDARDS FOR NEW SOURCES FOR THE PAVING AND ROOFING MATERIALS (TARS AND ASPHALT) POINT SOURCE CATEGORY Asphalt Emulsion Subcategory § 443.15 Standards of performance for new sources...

40 CFR 420.134 - New source performance standards (NSPS).

Code of Federal Regulations, 2010 CFR

2010-07-01

... Source Performance Standards (NSPS) Pollutant Maximum daily 1 Maximum monthly avg. 1 TSS 0.00998 0.00465... operations. Subpart M—New Source Performance Standards (NSPS) Pollutant Maximum daily 1 Maximum monthly avg...

40 CFR 420.134 - New source performance standards (NSPS).

Code of Federal Regulations, 2011 CFR

2011-07-01

... Source Performance Standards (NSPS) Pollutant Maximum daily 1 Maximum monthly avg. 1 TSS 0.00998 0.00465... operations. Subpart M—New Source Performance Standards (NSPS) Pollutant Maximum daily 1 Maximum monthly avg...

76 FR 52737 - Oil and Natural Gas Sector: New Source Performance Standards and National Emission Standards for...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-08-23

...This action announces how the EPA proposes to address the reviews of the new source performance standards for volatile organic compound and sulfur dioxide emissions from natural gas processing plants. We are proposing to add to the source category list any oil and gas operation not covered by the current listing. This action also includes proposed amendments to the existing new source performance standards for volatile organic compounds from natural gas processing plants and proposed standards for operations that are not covered by the existing new source performance standards. In addition, this action proposes how the EPA will address the residual risk and technology review conducted for the oil and natural gas production and natural gas transmission and storage national emission standards for hazardous air pollutants. This action further proposes standards for emission sources within these two source categories that are not currently addressed, as well as amendments to improve aspects of these national emission standards for hazardous air pollutants related to applicability and implementation. Finally, this action addresses provisions in these new source performance standards and national emission standards for hazardous air pollutants related to emissions during periods of startup, shutdown and malfunction.

40 CFR 432.15 - New source performance standards (NSPS).

Code of Federal Regulations, 2013 CFR

2013-07-01

... § 432.12(a)(1); and standards for ammonia (as N) are as follows: Performance Standards [NSPS] Regulatedparameter Maximum daily 1 Maximum monthly avg. 1 Ammonia (as N) 0.34 0.17 1 Pounds per 1000 lbs (or g/kg... ammonia (as N) apply: Performance Standards [NSPS] Regulatedparameter Maximum daily 1 Maximum monthly avg...

46 CFR 164.023-7 - Performance; non-standard thread.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 46 Shipping 6 2010-10-01 2010-10-01 false Performance; non-standard thread. 164.023-7 Section 164... Performance; non-standard thread. (a) Use Codes 1, 2, 3, 4BC, 4RB, 5 (any). Each non-standard thread which...) testing machine. (2) Single strand breaking strength (after weathering). After exposure in a sunshine...

46 CFR 164.023-7 - Performance; non-standard thread.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 46 Shipping 6 2011-10-01 2011-10-01 false Performance; non-standard thread. 164.023-7 Section 164... Performance; non-standard thread. (a) Use Codes 1, 2, 3, 4BC, 4RB, 5 (any). Each non-standard thread which...) testing machine. (2) Single strand breaking strength (after weathering). After exposure in a sunshine...

40 CFR 60.542a - Alternate standard for volatile organic compounds.

Code of Federal Regulations, 2010 CFR

2010-07-01

... PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for the Rubber Tire Manufacturing Industry § 60.542a Alternate standard for volatile organic compounds. (a) On and after the date on which the initial performance test, required by § 60.8, is completed, but no...

30 CFR 828.11 - In situ processing: Performance standards.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 30 Mineral Resources 3 2011-07-01 2011-07-01 false In situ processing: Performance standards. 828... STANDARDS-IN SITU PROCESSING § 828.11 In situ processing: Performance standards. (a) The person who conducts in situ processing activities shall comply with 30 CFR 817 and this section. (b) In situ processing...

30 CFR 828.11 - In situ processing: Performance standards.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 30 Mineral Resources 3 2012-07-01 2012-07-01 false In situ processing: Performance standards. 828... STANDARDS-IN SITU PROCESSING § 828.11 In situ processing: Performance standards. (a) The person who conducts in situ processing activities shall comply with 30 CFR 817 and this section. (b) In situ processing...

30 CFR 828.11 - In situ processing: Performance standards.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 30 Mineral Resources 3 2013-07-01 2013-07-01 false In situ processing: Performance standards. 828... STANDARDS-IN SITU PROCESSING § 828.11 In situ processing: Performance standards. (a) The person who conducts in situ processing activities shall comply with 30 CFR 817 and this section. (b) In situ processing...

30 CFR 828.11 - In situ processing: Performance standards.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 30 Mineral Resources 3 2010-07-01 2010-07-01 false In situ processing: Performance standards. 828... STANDARDS-IN SITU PROCESSING § 828.11 In situ processing: Performance standards. (a) The person who conducts in situ processing activities shall comply with 30 CFR 817 and this section. (b) In situ processing...

30 CFR 828.11 - In situ processing: Performance standards.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 30 Mineral Resources 3 2014-07-01 2014-07-01 false In situ processing: Performance standards. 828... STANDARDS-IN SITU PROCESSING § 828.11 In situ processing: Performance standards. (a) The person who conducts in situ processing activities shall comply with 30 CFR 817 and this section. (b) In situ processing...

40 CFR 443.35 - Standards of performance for new sources.

Code of Federal Regulations, 2011 CFR

2011-07-01

... PERFORMANCE AND PRETREATMENT STANDARDS FOR NEW SOURCES FOR THE PAVING AND ROOFING MATERIALS (TARS AND ASPHALT) POINT SOURCE CATEGORY Asphalt Roofing Subcategory § 443.35 Standards of performance for new sources. The...

40 CFR 443.35 - Standards of performance for new sources.

Code of Federal Regulations, 2010 CFR

2010-07-01

... PERFORMANCE AND PRETREATMENT STANDARDS FOR NEW SOURCES FOR THE PAVING AND ROOFING MATERIALS (TARS AND ASPHALT) POINT SOURCE CATEGORY Asphalt Roofing Subcategory § 443.35 Standards of performance for new sources. The...

30 CFR 828.1 - Scope.

Code of Federal Regulations, 2014 CFR

2014-07-01

... PERFORMANCE STANDARDS SPECIAL PERMANENT PROGRAM PERFORMANCE STANDARDS-IN SITU PROCESSING § 828.1 Scope. This part sets forth special environmental protection performance, reclamation and design standards for in situ processing activities. [44 FR 15455, Mar. 13, 1979] ...

30 CFR 828.1 - Scope.

Code of Federal Regulations, 2012 CFR

2012-07-01

... PERFORMANCE STANDARDS SPECIAL PERMANENT PROGRAM PERFORMANCE STANDARDS-IN SITU PROCESSING § 828.1 Scope. This part sets forth special environmental protection performance, reclamation and design standards for in situ processing activities. [44 FR 15455, Mar. 13, 1979] ...

30 CFR 828.1 - Scope.

Code of Federal Regulations, 2011 CFR

2011-07-01

... PERFORMANCE STANDARDS SPECIAL PERMANENT PROGRAM PERFORMANCE STANDARDS-IN SITU PROCESSING § 828.1 Scope. This part sets forth special environmental protection performance, reclamation and design standards for in situ processing activities. [44 FR 15455, Mar. 13, 1979] ...

30 CFR 828.1 - Scope.

Code of Federal Regulations, 2013 CFR

2013-07-01

... PERFORMANCE STANDARDS SPECIAL PERMANENT PROGRAM PERFORMANCE STANDARDS-IN SITU PROCESSING § 828.1 Scope. This part sets forth special environmental protection performance, reclamation and design standards for in situ processing activities. [44 FR 15455, Mar. 13, 1979] ...

Improving performance of Zambia Defence Force antiretroviral therapy providers: evaluation of a standards-based approach

PubMed Central

Kim, Young Mi; Banda, Joseph; Kanjipite, Webby; Sarkar, Supriya; Bazant, Eva; Hiner, Cyndi; Tholandi, Maya; Reinhardt, Stephanie; Njobvu, Panganani Dalisani; Kols, Adrienne; Benavides, Bruno

2013-01-01

ABSTRACT Background: The Zambia Defence Force (ZDF) has applied the Standards-Based Management and Recognition (SBM-R®) approach, which uses detailed performance standards, at some health facilities to improve HIV-related services offered to military personnel and surrounding civilian communities. This study examines the effectiveness of the SBM-R approach in improving facility readiness and provider performance at ZDF facilities. Methods: We collected data on facility readiness and provider performance before and after the 2010–2012 intervention at 4 intervention sites selected for their relatively poor performance and 4 comparison sites. Assessors observed whether each facility met 16 readiness standards and whether providers met 9 performance standards during consultations with 354 returning antiretroviral therapy (ART) clients. We then calculated the percentages of criteria achieved for each readiness and performance standard and conducted bivariate and multivariate analyses of provider performance data. Results: Facilities' ART readiness scores exceeded 80% before the intervention at both intervention and comparison sites. At endline, scores improved on 4 facility readiness standards in the intervention group but on only 1 standard in the comparison group. Multivariate analysis found that the overall provider performance score increased significantly in the intervention group (from 58% to 84%; P<.01) but not in the comparison group (from 62% to 70%). The before-and-after improvement in scores was significantly greater among intervention sites than among comparison sites for 2 standards—initial assessment of the client's condition and nutrition counseling. Conclusion: The standards-based approach, which involved intensive and mutually reinforcing intervention activities, showed modest improvements in some aspects of providers' performance during ART consultations. Further research is needed to determine whether improvements in provider performance affect client outcomes such as adherence to ART. PMID:25276534

Academy of nutrition and dietetics: revised 2014 standards of practice and standards of professional performance for registered dietitian nutritionists (competent, proficient, and expert) in sports nutrition and dietetics.

PubMed

Steinmuller, Patricia L; Kruskall, Laura J; Karpinski, Christine A; Manore, Melinda M; Macedonio, Michele A; Meyer, Nanna L

2014-04-01

Sports nutrition and dietetics addresses relationships of nutrition with physical activity, including weight management, exercise, and physical performance. Nutrition plays a key role in the prevention and treatment of obesity and chronic disease and for maintenance of health, and the ability to engage in physical activity, sports, and other aspects of physical performance. Thus, the Sports, Cardiovascular, and Wellness Nutrition Dietetic Practice Group, with guidance from the Academy of Nutrition and Dietetics Quality Management Committee, has developed the Revised 2014 Standards of Practice and Standards of Professional Performance as a resource for Registered Dietitian Nutritionists working in sports nutrition and dietetics to assess their current skill levels and to identify areas for further professional development in this emerging practice area. The revised document reflects advances in sports nutrition and dietetics practice since the original standards were published in 2009 and replaces those standards. The Standards of Practice represents the four steps in the Nutrition Care Process as applied to the care of patients/clients. The Standards of Professional Performance covers six standards of professional performance: quality in practice, competence and accountability, provision of services, application of research, communication and application of knowledge, and utilization and management of resources. Within each standard, specific indicators provide measurable action statements that illustrate how the standards can be applied to practice. The indicators describe three skill levels (competent, proficient, and expert) for Registered Dietitian Nutritionists working in sports nutrition and dietetics. The Standards of Practice and Standards of Professional Performance are complementary resources for Registered Dietitian Nutritionists in sports nutrition and dietetics practice. Copyright © 2014 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

Private and social costs of surface mine reforestation performance criteria.

PubMed

Sullivan, Jay; Amacher, Gregory S

2010-02-01

We study the potentially unnecessary costs imposed by strict performance standards for forest restoration of surface coal mines in the Appalachian region under the Surface Mining Control and Reclamation Act of 1977 (SMCRA) that can vary widely across states. Both the unnecessary private costs to the mine operator and costs to society (social costs) are reported for two performance standards, a ground cover requirement, and a seedling survival target. These standards are examined using numerical analyses under a range of site productivity class and market conditions. We show that a strict (90%) ground cover standard may produce an unnecessary private cost of more than $700/ha and a social cost ranging from $428/ha to $710/ha, as compared with a 70% standard. A strict tree survival standard of 1235 trees/ha, as compared with the more typical 1087 trees/ha standard, may produce an unnecessary private cost of approximately $200/ha, and a social cost in the range of $120 to $208/ha. We conclude that strict performance standards may impose substantial unnecessary private costs and social costs, that strict performance standards may be discouraging the choice of forestry as a post-mining land use, and that opportunities exist for reform of reforestation performance standards. Our study provides a basis for evaluating tradeoffs between regulatory efficiency and optimal reforestation effort.

40 CFR 447.15 - Standards of performance for new sources.

Code of Federal Regulations, 2012 CFR

2012-07-01

...) EFFLUENT GUIDELINES AND STANDARDS (CONTINUED) INK FORMULATING POINT SOURCE CATEGORY Oil-Base Solvent Wash Ink Subcategory § 447.15 Standards of performance for new sources. The following standards of...

40 CFR 447.15 - Standards of performance for new sources.

Code of Federal Regulations, 2014 CFR

2014-07-01

...) EFFLUENT GUIDELINES AND STANDARDS (CONTINUED) INK FORMULATING POINT SOURCE CATEGORY Oil-Base Solvent Wash Ink Subcategory § 447.15 Standards of performance for new sources. The following standards of...

40 CFR 447.15 - Standards of performance for new sources.

Code of Federal Regulations, 2013 CFR

2013-07-01

...) EFFLUENT GUIDELINES AND STANDARDS (CONTINUED) INK FORMULATING POINT SOURCE CATEGORY Oil-Base Solvent Wash Ink Subcategory § 447.15 Standards of performance for new sources. The following standards of...

40 CFR 427.85 - Standards of performance for new sources.

Code of Federal Regulations, 2010 CFR

2010-07-01

...) EFFLUENT GUIDELINES AND STANDARDS ASBESTOS MANUFACTURING POINT SOURCE CATEGORY Coating or Finishing of Asbestos Textiles Subcategory § 427.85 Standards of performance for new sources. The following standards of...

40 CFR 417.75 - Standards of performance for new sources.

Code of Federal Regulations, 2014 CFR

2014-07-01

...) EFFLUENT GUIDELINES AND STANDARDS SOAP AND DETERGENT MANUFACTURING POINT SOURCE CATEGORY Manufacture of Bar Soaps Subcategory § 417.75 Standards of performance for new sources. The following standards of...

40 CFR 417.85 - Standards of performance for new sources.

Code of Federal Regulations, 2014 CFR

2014-07-01

...) EFFLUENT GUIDELINES AND STANDARDS SOAP AND DETERGENT MANUFACTURING POINT SOURCE CATEGORY Manufacture of Liquid Soaps Subcategory § 417.85 Standards of performance for new sources. The following standards of...

40 CFR 417.75 - Standards of performance for new sources.

Code of Federal Regulations, 2010 CFR

2010-07-01

...) EFFLUENT GUIDELINES AND STANDARDS SOAP AND DETERGENT MANUFACTURING POINT SOURCE CATEGORY Manufacture of Bar Soaps Subcategory § 417.75 Standards of performance for new sources. The following standards of...

40 CFR 417.85 - Standards of performance for new sources.

Code of Federal Regulations, 2013 CFR

2013-07-01

...) EFFLUENT GUIDELINES AND STANDARDS SOAP AND DETERGENT MANUFACTURING POINT SOURCE CATEGORY Manufacture of Liquid Soaps Subcategory § 417.85 Standards of performance for new sources. The following standards of...

40 CFR 417.85 - Standards of performance for new sources.

Code of Federal Regulations, 2010 CFR

2010-07-01

...) EFFLUENT GUIDELINES AND STANDARDS SOAP AND DETERGENT MANUFACTURING POINT SOURCE CATEGORY Manufacture of Liquid Soaps Subcategory § 417.85 Standards of performance for new sources. The following standards of...

40 CFR 417.85 - Standards of performance for new sources.

Code of Federal Regulations, 2012 CFR

2012-07-01

...) EFFLUENT GUIDELINES AND STANDARDS SOAP AND DETERGENT MANUFACTURING POINT SOURCE CATEGORY Manufacture of Liquid Soaps Subcategory § 417.85 Standards of performance for new sources. The following standards of...

40 CFR 417.85 - Standards of performance for new sources.

Code of Federal Regulations, 2011 CFR

2011-07-01

...) EFFLUENT GUIDELINES AND STANDARDS SOAP AND DETERGENT MANUFACTURING POINT SOURCE CATEGORY Manufacture of Liquid Soaps Subcategory § 417.85 Standards of performance for new sources. The following standards of...

40 CFR 417.75 - Standards of performance for new sources.

Code of Federal Regulations, 2011 CFR

2011-07-01

...) EFFLUENT GUIDELINES AND STANDARDS SOAP AND DETERGENT MANUFACTURING POINT SOURCE CATEGORY Manufacture of Bar Soaps Subcategory § 417.75 Standards of performance for new sources. The following standards of...

40 CFR 417.75 - Standards of performance for new sources.

Code of Federal Regulations, 2013 CFR

2013-07-01

...) EFFLUENT GUIDELINES AND STANDARDS SOAP AND DETERGENT MANUFACTURING POINT SOURCE CATEGORY Manufacture of Bar Soaps Subcategory § 417.75 Standards of performance for new sources. The following standards of...

40 CFR 417.75 - Standards of performance for new sources.

Code of Federal Regulations, 2012 CFR

2012-07-01

...) EFFLUENT GUIDELINES AND STANDARDS SOAP AND DETERGENT MANUFACTURING POINT SOURCE CATEGORY Manufacture of Bar Soaps Subcategory § 417.75 Standards of performance for new sources. The following standards of...

40 CFR 446.15 - Standards of performance for new sources.

Code of Federal Regulations, 2013 CFR

2013-07-01

...) EFFLUENT GUIDELINES AND STANDARDS (CONTINUED) PAINT FORMULATING POINT SOURCE CATEGORY Oil-Base Solvent Wash Paint Subcategory § 446.15 Standards of performance for new sources. The following standards of...

40 CFR 446.15 - Standards of performance for new sources.

Code of Federal Regulations, 2012 CFR

2012-07-01

...) EFFLUENT GUIDELINES AND STANDARDS (CONTINUED) PAINT FORMULATING POINT SOURCE CATEGORY Oil-Base Solvent Wash Paint Subcategory § 446.15 Standards of performance for new sources. The following standards of...

40 CFR 446.15 - Standards of performance for new sources.

Code of Federal Regulations, 2014 CFR

2014-07-01

...) EFFLUENT GUIDELINES AND STANDARDS (CONTINUED) PAINT FORMULATING POINT SOURCE CATEGORY Oil-Base Solvent Wash Paint Subcategory § 446.15 Standards of performance for new sources. The following standards of...

78 FR 69928 - First Meeting: RTCA Special Committee 228-Minimum Operational Performance Standards for Unmanned...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-11-21

... 228--Minimum Operational Performance Standards for Unmanned Aircraft Systems AGENCY: Federal Aviation...--Minimum Operational Performance Standards for Unmanned Aircraft Systems. SUMMARY: The FAA is issuing this notice to advise the public of a meeting of RTCA Special Committee 228--Minimum Operational Performance...

78 FR 38093 - First Meeting: RTCA Special Committee 228-Minimum Operational Performance Standards for Unmanned...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-25

... 228--Minimum Operational Performance Standards for Unmanned Aircraft Systems AGENCY: Federal Aviation...--Minimum Operational Performance Standards for Unmanned Aircraft Systems. SUMMARY: The FAA is issuing this notice to advise the public of a meeting of RTCA Special Committee 228--Minimum Operational Performance...

Business Education. Vocational Education Program Courses Standards.

ERIC Educational Resources Information Center

Florida State Dept. of Education, Tallahassee. Div. of Vocational, Adult, and Community Education.

This document contains vocational education program courses standards (curriculum frameworks and student performance standards) for business technology education programs in Florida. Each program courses standard is composed of two parts: a curriculum framework and student performance standards. The curriculum framework includes four major…

40 CFR 60.62 - Standards.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 7 2013-07-01 2013-07-01 false Standards. 60.62 Section 60.62 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Portland Cement Plants § 60.62 Standards...

40 CFR 60.62 - Standards.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 7 2014-07-01 2014-07-01 false Standards. 60.62 Section 60.62 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Portland Cement Plants § 60.62 Standards...

40 CFR 60.174 - Standard for visible emissions.

Code of Federal Regulations, 2011 CFR

2011-07-01

... (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Primary Zinc... acid plant to comply with the standard set forth in § 60.173, any visible emissions which exhibit...

Ethics and choosing appropriate means to an end: problems with coal mine and nuclear workplace safety.

PubMed

Shrader-Frechette, Kristin; Cooke, Roger

2004-02-01

A common problem in ethics is that people often desire an end but fail to take the means necessary to achieve it. Employers and employees may desire the safety end mandated by performance standards for pollution control, but they may fail to employ the means, specification standards, necessary to achieve this end. This article argues that current (de jure) performance standards, for lowering employee exposures to ionizing radiation, fail to promote de facto worker welfare, in part because employers and employees do not follow the necessary means (practices known as specification standards) to achieve the end (performance standards) of workplace safety. To support this conclusion, the article argues that (1) safety requires attention to specification, as well as performance, standards; (2) coal-mine specification standards may fail to promote performance standards; (3) nuclear workplace standards may do the same; (4) choosing appropriate means to the end of safety requires attention to the ways uncertainties and variations in exposure may mask violations of standards; and (5) correcting regulatory inattention to differences between de jure and de facto is necessary for achievement of ethical goals for safety.

A Note on the Determination of "Acceptable" Performance in Thorndike's Standard of Fair Selection.

ERIC Educational Resources Information Center

Brown, Charles

1980-01-01

The determination of acceptable performance in Thorndike's constant ratio standard of fair selection is considered. It is shown that suitable choice of acceptable performance can make any minority-majority selection disparity consistent with Thorndike's standard. A rule for determining acceptable performance which avoids the Petersen-Novick…

49 CFR 179.400-4 - Insulation system and performance standard.

Code of Federal Regulations, 2012 CFR

2012-10-01

... of heat transfer used for determining the satisfactory performance of the insulation system of a... 49 Transportation 3 2012-10-01 2012-10-01 false Insulation system and performance standard. 179... DOT-113 and 107A) § 179.400-4 Insulation system and performance standard. (a) For the purposes of this...

49 CFR 179.400-4 - Insulation system and performance standard.

Code of Federal Regulations, 2010 CFR

2010-10-01

... of heat transfer used for determining the satisfactory performance of the insulation system of a... 49 Transportation 2 2010-10-01 2010-10-01 false Insulation system and performance standard. 179... DOT-113 and 107A) § 179.400-4 Insulation system and performance standard. (a) For the purposes of this...

49 CFR 179.400-4 - Insulation system and performance standard.

Code of Federal Regulations, 2013 CFR

2013-10-01

... of heat transfer used for determining the satisfactory performance of the insulation system of a... 49 Transportation 3 2013-10-01 2013-10-01 false Insulation system and performance standard. 179... DOT-113 and 107A) § 179.400-4 Insulation system and performance standard. (a) For the purposes of this...

49 CFR 179.400-4 - Insulation system and performance standard.

Code of Federal Regulations, 2011 CFR

2011-10-01

... of heat transfer used for determining the satisfactory performance of the insulation system of a... 49 Transportation 3 2011-10-01 2011-10-01 false Insulation system and performance standard. 179... DOT-113 and 107A) § 179.400-4 Insulation system and performance standard. (a) For the purposes of this...

49 CFR 179.400-4 - Insulation system and performance standard.

Code of Federal Regulations, 2014 CFR

2014-10-01

... of heat transfer used for determining the satisfactory performance of the insulation system of a... 49 Transportation 3 2014-10-01 2014-10-01 false Insulation system and performance standard. 179... DOT-113 and 107A) § 179.400-4 Insulation system and performance standard. (a) For the purposes of this...

Medicaid management information systems performance standards: Health Care Financing Administration. Notice with comment period.

PubMed

1981-06-30

This notice contains performance standards (review elements and factors). We are required by section 1903(r)(6)(E) of the Social Security Act to notify all States of proposed procedures, standards, and other requirements at least one quarter prior to the fiscal year in which the procedures, standards, and other requirements will be used for Medicaid Management Information Systems reapproval reviews. This Notice meets that statutory requirements. By October 1, 1981, we will use the performance standards and existing systems requirements when conducting the annual review of State system performance.

21 CFR 861.30 - Development of standards.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Development of standards. 861.30 Section 861.30...) MEDICAL DEVICES PROCEDURES FOR PERFORMANCE STANDARDS DEVELOPMENT Procedures for Performance Standards Development and Publication § 861.30 Development of standards. The Food and Drug Administration (FDA), while...

21 CFR 861.30 - Development of standards.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Development of standards. 861.30 Section 861.30...) MEDICAL DEVICES PROCEDURES FOR PERFORMANCE STANDARDS DEVELOPMENT Procedures for Performance Standards Development and Publication § 861.30 Development of standards. The Food and Drug Administration (FDA), while...

40 CFR 464.44 - New source performance standards.

Code of Federal Regulations, 2010 CFR

2010-07-01

..., total phenols, oil and grease, and TSS. For non-continuous dischargers, annual average mass standards.... Concentration standards and annual average mass standards shall only apply to non-continuous dischargers. (a... 40 Protection of Environment 29 2010-07-01 2010-07-01 false New source performance standards. 464...

40 CFR 464.14 - New source performance standards.

Code of Federal Regulations, 2013 CFR

2013-07-01

..., total phenols, oil and grease, and TSS. For non-continuous dischargers, annual average mass standards.... Concentration standards and annual average mass standards shall only apply to non-continuous dischargers. (a... 40 Protection of Environment 31 2013-07-01 2013-07-01 false New source performance standards. 464...

40 CFR 464.14 - New source performance standards.

Code of Federal Regulations, 2010 CFR

2010-07-01

..., total phenols, oil and grease, and TSS. For non-continuous dischargers, annual average mass standards.... Concentration standards and annual average mass standards shall only apply to non-continuous dischargers. (a... 40 Protection of Environment 29 2010-07-01 2010-07-01 false New source performance standards. 464...

40 CFR 464.44 - New source performance standards.

Code of Federal Regulations, 2014 CFR

2014-07-01

..., total phenols, oil and grease, and TSS. For non-continuous dischargers, annual average mass standards.... Concentration standards and annual average mass standards shall only apply to non-continuous dischargers. (a... 40 Protection of Environment 30 2014-07-01 2014-07-01 false New source performance standards. 464...

40 CFR 464.24 - New source performance standards.

Code of Federal Regulations, 2013 CFR

2013-07-01

..., total phenols, oil and grease, and TSS. For non-continuous dischargers, annual average mass standards.... Concentration standards and annual average mass standards shall only apply to non-continuous dischargers. (a... 40 Protection of Environment 31 2013-07-01 2013-07-01 false New source performance standards. 464...

40 CFR 464.24 - New source performance standards.

Code of Federal Regulations, 2011 CFR

2011-07-01

..., total phenols, oil and grease, and TSS. For non-continuous dischargers, annual average mass standards.... Concentration standards and annual average mass standards shall only apply to non-continuous dischargers. (a... 40 Protection of Environment 30 2011-07-01 2011-07-01 false New source performance standards. 464...

40 CFR 464.24 - New source performance standards.

Code of Federal Regulations, 2010 CFR

2010-07-01

..., total phenols, oil and grease, and TSS. For non-continuous dischargers, annual average mass standards.... Concentration standards and annual average mass standards shall only apply to non-continuous dischargers. (a... 40 Protection of Environment 29 2010-07-01 2010-07-01 false New source performance standards. 464...

40 CFR 464.44 - New source performance standards.

Code of Federal Regulations, 2013 CFR

2013-07-01

..., total phenols, oil and grease, and TSS. For non-continuous dischargers, annual average mass standards.... Concentration standards and annual average mass standards shall only apply to non-continuous dischargers. (a... 40 Protection of Environment 31 2013-07-01 2013-07-01 false New source performance standards. 464...

40 CFR 464.44 - New source performance standards.

Code of Federal Regulations, 2012 CFR

2012-07-01

..., total phenols, oil and grease, and TSS. For non-continuous dischargers, annual average mass standards.... Concentration standards and annual average mass standards shall only apply to non-continuous dischargers. (a... 40 Protection of Environment 31 2012-07-01 2012-07-01 false New source performance standards. 464...

40 CFR 464.24 - New source performance standards.

Code of Federal Regulations, 2014 CFR

2014-07-01

..., total phenols, oil and grease, and TSS. For non-continuous dischargers, annual average mass standards.... Concentration standards and annual average mass standards shall only apply to non-continuous dischargers. (a... 40 Protection of Environment 30 2014-07-01 2014-07-01 false New source performance standards. 464...

40 CFR 464.44 - New source performance standards.

Code of Federal Regulations, 2011 CFR

2011-07-01

..., total phenols, oil and grease, and TSS. For non-continuous dischargers, annual average mass standards.... Concentration standards and annual average mass standards shall only apply to non-continuous dischargers. (a... 40 Protection of Environment 30 2011-07-01 2011-07-01 false New source performance standards. 464...

40 CFR 464.24 - New source performance standards.

Code of Federal Regulations, 2012 CFR

2012-07-01

..., total phenols, oil and grease, and TSS. For non-continuous dischargers, annual average mass standards.... Concentration standards and annual average mass standards shall only apply to non-continuous dischargers. (a... 40 Protection of Environment 31 2012-07-01 2012-07-01 false New source performance standards. 464...

40 CFR 464.14 - New source performance standards.

Code of Federal Regulations, 2014 CFR

2014-07-01

..., total phenols, oil and grease, and TSS. For non-continuous dischargers, annual average mass standards.... Concentration standards and annual average mass standards shall only apply to non-continuous dischargers. (a... 40 Protection of Environment 30 2014-07-01 2014-07-01 false New source performance standards. 464...

40 CFR 464.14 - New source performance standards.

Code of Federal Regulations, 2011 CFR

2011-07-01

..., total phenols, oil and grease, and TSS. For non-continuous dischargers, annual average mass standards.... Concentration standards and annual average mass standards shall only apply to non-continuous dischargers. (a... 40 Protection of Environment 30 2011-07-01 2011-07-01 false New source performance standards. 464...

40 CFR 464.14 - New source performance standards.

Code of Federal Regulations, 2012 CFR

2012-07-01

..., total phenols, oil and grease, and TSS. For non-continuous dischargers, annual average mass standards.... Concentration standards and annual average mass standards shall only apply to non-continuous dischargers. (a... 40 Protection of Environment 31 2012-07-01 2012-07-01 false New source performance standards. 464...

40 CFR 60.253 - Standards for pneumatic coal-cleaning equipment.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 40 Protection of Environment 7 2013-07-01 2013-07-01 false Standards for pneumatic coal-cleaning... PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Coal Preparation and Processing Plants § 60.253 Standards for pneumatic coal-cleaning equipment. (a) On and after...

40 CFR 60.253 - Standards for pneumatic coal-cleaning equipment.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 40 Protection of Environment 6 2011-07-01 2011-07-01 false Standards for pneumatic coal-cleaning... PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Coal Preparation and Processing Plants § 60.253 Standards for pneumatic coal-cleaning equipment. (a) On and after...

40 CFR 60.253 - Standards for pneumatic coal-cleaning equipment.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 40 Protection of Environment 7 2012-07-01 2012-07-01 false Standards for pneumatic coal-cleaning... PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Coal Preparation and Processing Plants § 60.253 Standards for pneumatic coal-cleaning equipment. (a) On and after...

40 CFR 60.253 - Standards for pneumatic coal-cleaning equipment.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 40 Protection of Environment 6 2010-07-01 2010-07-01 false Standards for pneumatic coal-cleaning... PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Coal Preparation and Processing Plants § 60.253 Standards for pneumatic coal-cleaning equipment. (a) On and after...

40 CFR 60.253 - Standards for pneumatic coal-cleaning equipment.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 40 Protection of Environment 7 2014-07-01 2014-07-01 false Standards for pneumatic coal-cleaning... PROGRAMS (CONTINUED) STANDARDS OF PERFORMANCE FOR NEW STATIONARY SOURCES Standards of Performance for Coal Preparation and Processing Plants § 60.253 Standards for pneumatic coal-cleaning equipment. (a) On and after...