Structure-function studies of the plant cyclotides: The role of a circular protein backbone

The traditional idea of proteins as linear chains of amino acids is being challenged with the discovery of miniproteins that contain a circular backbone. The cyclotide family is the largest group of circular proteins and is characterized by an amide-circularized protein backbone and six conserved cysteine residues. These conserved cysteines are paired to form a knotted network of disulfide bonds. The combination of the circular backbone and a cystine knot, known as the cyclic cystine knot (CCK) motif, confers exceptional stability upon the cyclotides. This review discusses the role of the circular backbone based on studies of both the oxidative folding of kalata B1, the prototypical cyclotide, and a comparison of the structure and activity of kalata B1 and its acyclic permutants.

Equilibrium swelling measurements of network and semicrystalline polymers in supercritical carbon dioxide using high-pressure NMR

The extent of swelling of cross-linked poly(dimethylsiloxane) and linear low-density poly(ethylene) in supercritical CO2 has been investigated using high-pressure NMR spectroscopy and microscopy. Poly(dimethylsiloxane) was cross-linked to four different cross-link densities and swollen in supercritical CO2. The Flory-Huggins interaction parameter, x, was found to be 0.62 at 300 bar and 45 degrees C, indicating that supercritical CO2 is a relatively poor solvent compared to toluene or benzene. Linear low-density poly(ethylene) was shown to exhibit negligible swelling upon exposure to supercritical CO2 up to 300 bar. The effect Of CO2 pressure on the amorphous region of the poly(ethylene) was investigated by observing changes in the H-1 T-2 relaxation times of the polymer. These relaxation times decreased with increasing pressure, which was attributed to a decrease in mobility of the polymer chains as a result of compressive pressure.

Segmented polyurethane nanocomposites: Impact of controlled particle size nanofillers on the morphological response to uniaxial deformation

A series of TPU nanocomposites were prepared by incorporating organically modified layered silicates with controlled particle size. To our knowledge, this is the first study into the effects of layered silicate diameter in polymer nanocomposites utilizing the same mineral for each size fraction. The tensile properties of these materials were found to be highly dependent upon the size of the layered silicates. A decrease in disk diameter was associated with a sharp upturn in the stress-strain curve and a pronounced increase in tensile strength. Results from SAXS/SANS experiments showed that the layered silicates did not affect the bulk TPU microphase structure and the morphological response of the host TPU to deformation or promote/hinder strain-induced soft segment crystallization. The improved tensile properties of the nanocomposites containing the smaller nanofillers resulted from the layered silicates aligning in the direction of strain and interacting with the TPU sequences via secondary bonding. This phenomenon contributes predominantly above 400% strain once the microdomain architecture has largely been disassembled. Large tactoids that are unable to align in the strain direction lead to concentrated tensile stresses between the polymer and filler, instead of desirable shear stresses, resulting in void formation and reduced tensile properties. In severe cases, such as that observed for the composite containing the largest silicate, these voids manifest visually as stress whitening.

Recombinant human activated protein C improves pulmonary function in ovine acute lung injury resulting from smoke inhalation and sepsis

Objective: To investigate the effects of recombinant human activated protein C (rhAPC) on pulmonary function in acute lung injury (ALI) resulting from smoke inhalation in association with a bacterial challenge. Design: Prospective, randomized, controlled, experimental animal study with repeated measurements. Setting: Investigational intensive care unit at a university hospital. Subjects: Eighteen sheep (37.2 +/- 1.0 kg) were operatively prepared and randomly allocated to either the sham, control, or rhAPC group (n = 6 each). After a tracheotomy had been performed, ALI was produced in the control and rhAPC group by insufflation of 4 sets of 12 breaths of cotton smoke. Then, a 30 mL suspension of live Pseudomonas aeruginosa bacteria (containing 2-5 x 10(11) colony forming units) was instilled into the lungs according to an established protocol. The sham group received only the vehicle, i.e., 4 sets of 12 breaths of room air and instillation of 30 mL normal saline. The sheep were studied in the awake state for 24 hrs and were ventilated with 100% oxygen. RhAPC (24 mu g/kg/hr) was intravenously administered. The infusion was initiated 1 hr post-injury and lasted until the end of the experiment. The animals were resuscitated with Ringer's lactate solution to maintain constant pulmonary artery occlusion pressure. Measurements and Main Results., In comparison with nontreatment in controls, the infusion of rhAPC significantly attenuated the fall in PaO2/FiO(2) ratio (control group values were 521 +/- 22 at baseline [BL], 72 +/- 5 at 12 hrs, and 74 +/- 7 at 24 hrs, vs. rhAPC group values of 541 +/- 12 at BL, 151 +/- 29 at 12 hours [p < .05 vs. control], and 118 +/- 20 at 24 hrs), and significantly reduced the increase in pulmonary microvascular shunt fraction (Qs/Qt; control group at BL, 0.14 +/- 0.02, and at 24 hrs, 0.65 +/- 0.08; rhAPC group at BL, 0.24 +/- 0.04, and at 24 hrs, 0.45 +/- 0.02 [p < .05 vs. control]) and the increase in peak airway pressure (mbar; control group at BL, 20 +/- 1, and at 24 hrs, 36 +/- 4; rhAPC group at BL, 21 +/- 1, and at 24 hrs, 28 +/- 2 [p < .05 vs. control]). In addition, rhAPC limited the increase in lung 3-nitrotyrosine (after 24 hrs [%]: sham, 7 +/- 2; control, 17 +/- 1; rhAPC, 12 +/- 1 [p < .05 vs. control]), a reliable indicator of tissue injury. However, rhAPC failed to prevent lung edema formation. RhAPC-treated sheep showed no difference in activated clotting time or platelet count but exhibited less fibrin degradation products (1/6 animals) than did controls (4/6 animals). Conclusions. Recombinant human activated protein C attenuated ALI after smoke inhalation and bacterial challenge in sheep, without bleeding complications.

Development of the Minimum Information Specification for in situ Hybridization and Immunohistochemistry Experiments (MISFISHIE)

We describe the creation process of the Minimum Information Specification for In Situ Hybridization and Immunohistochemistry Experiments (MISFISHIE). Modeled after the existing minimum information specification for microarray data, we created a new specification for gene expression localization experiments, initially to facilitate data sharing within a consortium. After successful use within the consortium, the specification was circulated to members of the wider biomedical research community for comment and refinement. After a period of acquiring many new suggested requirements, it was necessary to enter a final phase of excluding those requirements that were deemed inappropriate as a minimum requirement for all experiments. The full specification will soon be published as a version 1.0 proposal to the community, upon which a more full discussion must take place so that the final specification may be achieved with the involvement of the whole community. This paper is part of the special issue of OMICS on data standards.

Simulation of glassy state relaxations in polymers: A static analysis of methyl group and methoxy group rotation in poly(vinyl methyl ether)

We show that the simple quasi-static technique, also called the adiabatic mapping technique, can be used to determine the energetics of rotation of methyl and methoxy groups in amorphous poly(vinyl methyl ether) even though the latter process is too slow to be amenable to direct molecular dynamics simulation. For the methyl group rotation, we find that the mean and standard deviation of the simulated rotational barrier heights agree well with experimental data from quasi-elastic neutron scattering. In the case of the methoxy groups we find that just 4% of the groups contribute more than 90% of the observed dielectric relaxation strength. The groups which make the most contribution are those which, by virtue of their particular conformation and local environment, have two alternative positions of similar energy.

Solution properties of star and linear poly(N-isopropylacrylamide)

The LCST transitions of novel N-isopropylacrylamide ( NIPAM) star polymers, prepared using the four-armed RAFT agent pentaerythritoltetrakis(3-(S-benzyltrithiocarbonyl) propionate) (PTBTP) and their hydrolyzed linear arms were studied using H-1 NMR, PFG-NMR, and DLS. The aim was to determine the effect of polymer architecture and the presence of end groups derived from RAFT agents on the LCST. The LCST transitions of star PNIPAM were significantly depressed by the presence of the hydrophobic star core and possibly the benzyl end groups. The effect was molecular weight dependent and diminished once the number of repeating units per arm >= 70. The linear PNIPAM exhibited an LCST of 35 degrees C, regardless of molecular weight; the presence of both hydrophilic and hydrophobic end groups after hydrolysis from the star core was suggested to cancel effects on the LCST. A significant decrease in R-H was observed below the LCST for star and linear PNIPAM and was attributed to the formation of n-clusters. Application of a scaling law to the linear PNIPAM data indicated the cluster size n = 6. Tethering to the hydrophobic star core appeared to inhibit n-cluster formation in the lowest molecular weight stars; this may be due to enhanced stretching of the polymer chains, or the presence of larger numbers of n-clusters at temperatures below those measured.

DOSY NMR studies of chemical exchange behavior of poly(2-hydroxyethyl methacrylate)

PFG-NMR was used to study the chemical exchange of linear PHEMA having a range of molecular weights with water in DMSO containing varying quantities of water. The aim was to investigate the use of PFG-NMR to study chemical exchange between a polymer with exchangeable protons and a small fast diffusing molecule to provide insight into the conformation adopted by a polymer in solution. The experimental data were simulated closely for the two-site exchange case using the Bloch equations modified for chemical exchange and diffusion. The exchange rate could be used to detect changes in polymer conformation resulting from changes in the solvent. PHEMA of MW 10 000 showed significant time-dependent changes in exchange rate, resulting from preferential solvation of the OH sites by water, and subsequent conformational changes which altered accessibility of the OH sites to water. This behavior was not observed for larger MW PHEMA, which adopted a stable conformation immediately. Large changes in the exchange rate were not reflected in changes to the hydrodynamic radius, suggesting that a minimal overall change in the chain dimensions occurred. DMSO was found to be a poor solvent for PHEMA, which adopts a compact conformation in DMSO. This work has demonstrated that PFG-NMR is a sensitive method for detecting subtle changes in polymer conformation in polymers with exchangeable protons.

Measuring and predicting the dynamics of linear monodisperse entangled polymers in rapid flow through an abrupt contraction. A small angle neutron scattering study

Small-angle neutron scattering measurements on a series of monodisperse linear entangled polystyrene melts in nonlinear flow through an abrupt 4:1 contraction have been made. Clear signatures of melt deformation and subsequent relaxation can be observed in the scattering patterns, which were taken along the centerline. These data are compared with the predictions of a recently derived molecular theory. Two levels of molecular theory are used: a detailed equation describing the evolution of molecular structure over all length scales relevant to the scattering data and a simplified version of the model, which is suitable for finite element computations. The velocity field for the complex melt flow is computed using the simplified model and scattering predictions are made by feeding these flow histories into the detailed model. The modeling quantitatively captures the full scattering intensity patterns over a broad range of data with independent variation of position within the contraction geometry, bulk flow rate and melt molecular weight. The study provides a strong, quantitative validation of current theoretical ideas concerning the microscopic dynamics of entangled polymers which builds upon existing comparisons with nonlinear mechanical stress data. Furthermore, we are able to confirm the appreciable length scale dependence of relaxation in polymer melts and highlight some wider implications of this phenomenon.

VALORAÇÃO DE ATIVOS E MULTAS AMBIENTAIS: ANÁLISE SIMULADA DE EMPREENDIMENTOS HOTELEIROS EM ÁREAS PROTEGIDAS

O crescimento desordenado e a ausência de políticas públicas mais eficientes levaram a uma diversificação quanto ao uso dos recursos naturais, principalmente no que se refere à água para o saneamento básico. O marco regulatório estabelecido na década de 1980 registrou as políticas públicas para o meio ambiente, que trouxeram avanços sobre o tema, desencadeando uma série de ações voltadas tanto para a estrutura burocrática e da prevenção, quanto solução para os problemas de degradação e esgotamento dos recursos naturais. Com o advento das leis específicas de proteção aos mananciais e mediante a lei 13.579/09 do Estado de São Paulo que trata sobre a área da Billings, percebeu-se um avanço na questão do gerenciamento para proteção e desenvolvimento de acordo com as características da região. Instrumentos de políticas públicas para conter as ações referentes aos danos causados ao meio ambiente, como a lei contra crimes ambientais foram às ações práticas do Estado para conter tais ações. O Objetivo desta pesquisa é analisar as possíveis discrepâncias entre as penalidades financeiras aplicadas na ocorrência das infrações ambientais e os modelos de valoração dos ativos ambientais, utilizando a simulação de implantação de um hotel em áreas de proteção e recuperação de manancial no Município de São Bernardo do Campo no Estado de São Paulo. O desenvolvimento da pesquisa se baseou no método de custo de reposição (MCR) para dimensionar os possíveis impactos gerados por um empreendimento hoteleiro e seu respectivo valor econômico. Posteriormente, os impactos ambientais foram relacionados com a legislação do município para determinar os valores das possíveis penalidades aplicáveis ao dano causado. Dentre os resultados levantados, verificou-se uma significativa discrepância entre a valoração econômica e as multas aplicáveis, sendo que nos impactos referentes a impermeabilidade do solo e contaminação do lençol freático, com diferenças superiores em relação às penalidades de R$ 804.922,78 e R$ 453.333,33 respectivamente. A partir da metodologia aplicada na pesquisa, observou-se que as penalidades incidentes em casos de danos ao meio ambiente, muitas vezes não atinge o objetivo, que é inibir a ação do infrator, pois o real custo econômico não é medido na aplicação do valor da multa.

O MERCADO DE TRABALHO PARA O PROFISSIONAL DA HOTELARIA NAS CIDADES DE GUARULHOS E SÃO BERNARDO DO CAMPO

O objetivo desse estudo é descobrir se as cidades de Guarulhos e São Bernardo do Campo têm chances de, ao explorar sua vocação turística para os negócios, obter reais possibilidades de ampliar seu mercado de trabalho na área de hotelaria. Para realizar o que se propôs, utilizou-se dos dados primários advindos das entrevistas pessoais com alguns profissionais ligados ao setor; além dos dados secundários e informações coletadas nos próprios sites das prefeituras, sindicatos, meios de hospedagem e outras associações que integrados à pesquisa bibliográfica verificou-se que as duas cidades possuem potencial turístico para os negócios porque contam com parques industriais que possibilitam a entrada e permanência de pessoas que vem às cidades para realizar diversos tipos de negócios. Para tanto, precisam hospedar-se, geralmente, por um espaço de tempo em que pretendem descansar, alimentar-se, banhar-se como se estivessem em suas casas. Além desse potencial, as cidades apresentam mais algumas características em comum como a localização de aeroportos, Cumbica e Congonhas, bem próximos de seus centros e, ainda, servem de passagem para outras cidades consideradas mais acolhedoras como Santos, Guarujá entre outras localizadas no litoral e São Paulo, na região conhecida como a grande São Paulo. Por isso Guarulhos e São Bernardo acabam sendo pouco aproveitadas no que se refere aos seus potenciais turísticos. A pesquisa aponta que se houver investimentos nesse setor, o turismo poderá crescer tornando-as cidades receptivas àqueles que as visitam por motivo de trabalho e que precisam de uma boa hospedagem e, muitas vezes, de um lugar para utilizar como seu próprio escritório. Com isso, aumenta-se a necessidade de profissionais qualificados para atender às expectativas desses turistas. Conseqüentemente amplia-se o mercado de trabalho nas áreas voltadas ao turismo e hotelaria das cidades. Entre elas, destacam-se Transportes, Comércio, Segurança, Gastronomia, Cultura, Esportes e Lazer, Redes Hoteleiras e até mesmo Escolas Técnicas e Faculdades. Identificou-se que as duas cidades estudadas possuem potencial turístico para os negócios e podem ampliar o mercado de trabalho hoteleiro a partir da conscientização das partes interessadas em investirem na infra-estrutura, na capacitação dos Recursos Humanos, na estrutura dos empreendimentos hoteleiros e na divulgação do local, dos produtos e dos serviços oferecidos.(AU)

A linguistic resurrection? Aller + infinitive in narrative past contexts

This article studies the go periphrasis emerging in Contemporary French narrations and compares it with the narrative go periphrasis found in Middle French.