Interaction of giant extracellular Glossoscolex paulistus hemoglobin (HbGp) with zwitterionic surfactant N-hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (HPS): Effects of oligomeric dissociation

The present work focuses on the interaction between the zwitterionic surfactant N-hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (HPS) and the giant extracellular hemoglobin of Glossoscolex paulistus (HbGp). Electronic optical absorption, fluorescence emission and circular dichroism spectroscopy techniques, together with Gel-filtration chromatography, were used in order to evaluate the oligomeric dissociation as well as the autoxidation of HbGp as a function of the interaction with HPS. A peculiar behavior was observed for the HPS-HbGp interaction: a complex ferric species formation equilibrium was promoted, as a consequence of the autoxidation and oligomeric dissociation processes. At pH 7.0, HPS is more effective up to 1 mM while at pH 9.0 the surfactant effect is more intense above 1 mM. Furthermore, the interaction of HPS with HbGp was clearly less intense than the interaction of this hemoglobin with cationic (CTAC) and anionic (SDS) surfactants. Probably, this lower interaction with HPS is due to two factors: (i) the lower electrostatic attraction between the HPS surfactant and the protein surface ionic sites when compared to the electrostatic interaction between HbGp and cationic and anionic surfactants, and (ii) the low cmc of HPS, which probably reduces the interaction of the surfactant in the monomeric form with the protein. The present work emphasizes the importance of the electrostatic contribution in the interaction between ionic surfactants and HbGp. Furthermore, in the whole HPS concentration range used in this study, no folding and autoxidation decrease induced by this surfactant were observed. This is quite different from the literature data on the interaction between surfactants and tetrameric hemoglobins, that supports the occurrence of this behavior for the intracellular hemoglobins at low surfactant concentration range. Spectroscopic data are discussed and compared with the literature in order to improve the understanding of hemoglobin-surfactant interaction as well as the acid isoelectric point (pI) influence of the giant extracellular hemoglobins on their structure-activity relationship. (c) 2007 Elsevier B.V. All rights reserved.

Interaction of bovine serum albumin (BSA) with ionic surfactants evaluated by electron paramagnetic resonance (EPR) spectroscopy

EPR spectra of 5- and 16-doxyl stearic acid nitroxide probes (5-DSA and 16-DSA, respectively) bound to bovine serum albumin (BSA) revealed that in the presence of ionic surfactants, at least, two label populations coexist in equilibrium. The rotational correlation times (tau) indicated that component I displays a more restricted mobility state, associated to the spin labels bound to the protein; the less immobilized component 2 is due to label localization in the surfactant aggregates. For both probes, the increase of surfactant concentration leads to higher motional levels of component 1 followed by a simultaneous decrease of this fraction of nitroxides and its conversion into component 2. For 10 mM cethyltrimethylammonium chloride (CTAC), the nitroxides are 100% bound to the protein, whereas at 10mM N-hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (HPS) and sodium dodecyl sulfate (SDS) the fractions of bound nitroxides are reduced to 18% and 86%, respectively. No significant polarity changes were observed in the whole surfactant concentration range for component 1. Moreover, at higher surfactant concentration, component 2 exhibited a similar polarity as in the pure surfactant micelles. For 16-DSA the surfactant effect is different: at 10mM of HPS and CTAC the fractions of bound nitroxides are 76% and 49%, respectively, while at 10 mM SDS they are present exclusively in a micellar environment, consistent with 100% of component 2. Overall, both SDS and HPS are able to effectively displace the nitroxide probes from the protein binding sites. while CTAC seems to affect the nitroxide binding to a significantly smaller extent. (C) 2008 Elsevier B.V. All rights reserved.

Microwave-assisted digestion procedures for biological samples with diluted nitric acid: Identification of reaction products

Microwave-assisted sample preparation using diluted nitric acid solutions is an alternative procedure for digesting organic samples. The efficiency of this procedure depends on the chemical properties of the samples and in this work it was evaluated by the determination of crude protein amount. fat and original carbon. Soybeans grains, bovine blood. bovine muscle and bovine viscera were digested in a cavity-microwave oven using oxidant mixtures in different acid concentrations. The digestion efficiency was evaluated based on the determination of residual carbon content and element recoveries using inductively coupled plasma optical emission spectrometry (ICP OES). In order to determine the main residual organic compounds, the digests were characterized by nuclear magnetic resonance (1 H NMR). Subsequently, studies concerning separation of nitrobenzoic acid isomers were performed by ion pair reversed phase liquid chromatography using a C18 stationary phase, water:acetonitrile:methanol (75:20:5, v/v/v) +0.05% (v/v) TFA as mobile phase and ultraviolet detection at 254 nm. Sample preparation based on diluted acids proved to be feasible and a recommendable alternative for organic sample digestion, reducing both the reagent volumes and the variability of the residues as a result of the process of decomposition. It was shown that biological matt-ices containing amino acids, proteins and lipids in their composition produced nitrobenzoic acid isomers and other organic compounds after cleavage of chemical bonds. (C) 2009 Elsevier B.V. All rights reserved.

Breaking the Conventions : Shakespeare, the Fair Young Man and the Dark Lady

The aim of this essay is to show how Shakespeare’s sonnets violated and reversed the conventional ideas in terms of beauty and idealisation. Furthermore, I will examine the way Shakespeare presented his beloved woman as an absolute opposite of the one created by Petrarch, and how he shifted all the divine metaphors from a woman to a fair young man, creating a new object of praise and admiration.

Elevers uppfattningar av vad man bör uppnå efter avslutad kurs i Idrott och Hälsa A : En studie av två gymnasieskolor i årskurs 2 – ett elevperspektiv

The purpose of this work is to see if the students, on two separate high schools, understand what skills you should achieve after completing the course in physical education and health A 100 points. The entire study is based on a student perspective. In order to best answer the purpose, the questionnaire used in the "available groups" in schools. The questions for the work are: To what extent and how students perceive what they should achieve after completing the course in Sport and Health A?, Which is the goal, based on the curriculum in Physical Education, that students feel are important to achieve? and how do a comparison like between these two schools on the basis of what students perceive that they must achieve in Sport and Health A?The result shows that the main goal that finds support in the course objectives is an assertion; Has knowledge of what to eat to maintain or improve health, which over 90% of students had checked in. The least important goal was the claim 14; Know woods and fields and carry out outdoor activities, as 30% of students had checked in. The most important goal that does not find support in the course objectives is claim 10; to be reversed to the lessons, which was 88% in response rate among students. Here was the least important goal, claim 6; to be skilled in various ball games, which received 17% of student responses. The findings revealed that there is a clear health perspective in physical education in the studied groups at the two schools, one can also see that there is an uneven distribution of the elements included in the curriculum of Sport and Health A, and the outdoor life and dance is rare in the teaching of the groups studied at the two schools. Finally, it appeared that the lesson content and teaching are likely to have a significant role in student perceptions of course goals.

Who Is the Shrew? : Irony as deconstruction in The Taming of the Shrew

In this thesis William Shakespeare’s comedy The Taming of The Shrew is analyzed from a feminist perspective. It is argued that the female characters Katherina and Bianca are mainly constructed through language expressed by male characters as binary oppositions. By an act of deconstruction within the play, these images, or constructed representations, are adjusted and somewhat reversed in the end. In addition, it is shown how the Induction, an introductory part of the play, adds aspects of construction and deconstruction, which supports an ironic reading and a questioning of the constructed gender roles.