Immobilized analogues of sunflower trypsin inhibitor-1 constitute a versatile group of affinity sorbents for selective isolation of serine proteases

Sunflower trypsin inhibitor-1 (SFI-1), a natural 14-residue cyclic peptide, and some of its synthetic acyclic variants are potent protease inhibitors displaying peculiar inhibitory profiles. Here we describe the synthesis and use of affinity sorbents prepared by coupling SFTI-1 analogues to agarose resin. Chymotrypsinand trypsin-like proteases could then be selectively isolated from pancreatin; similarly, other proteases were obtained from distinct biological sources. The binding capacity of [Lys5]-SFTI-1-agarose for trypsin was estimated at over 10 mg/mL of packed gel. SFTI-1-based resins could find application either to improve the performance of current purification protocols or as novel protease-discovery tools in different areas of biological investigation. (C) 2009 Elsevier B.V. All rights reserved.

Sample stacking in CZE using dynamic thermal junctions I. Analytes with low dpK(a)/dT crossing a single thermally induced pH junction in a BGE with high dpH/dT

The possibility to compress analyte bands at the beginning of CE runs has many advantages. Analytes at low concentration can be analyzed with high signal-to-noise ratios by using the so-called sample stacking methods. Moreover, sample injections with very narrow initial band widths (small initial standard deviations) are sometimes useful, especially if high resolutions among the bands are required in the shortest run time. In the present work, a method of sample stacking is proposed and demonstrated. It is based on BGEs with high thermal sensitive pHs (high dpH/dT) and analytes with low dpK(a)/dT. High thermal sensitivity means that the working pK(a) of the BGE has a high dpK(a)/dT in modulus. For instance, Tris and Ethanolamine have dpH/dT = -0.028/degrees C and -0.029/degrees C, respectively, whereas carboxylic acids have low dpK(a)/dT values, i.e. in the -0.002/degrees C to+0.002/degrees C range. The action of cooling and heating sections along the capillary during the runs affects also the local viscosity, conductivity, and electric field strength. The effect of these variables on electrophoretic velocity and band compression is theoretically calculated using a simple model. Finally, this stacking method was demonstrated for amino acids derivatized with naphthalene-2,3-dicarboxaldehyde and fluorescamine using a temperature difference of 70 degrees C between two neighbor sections and Tris as separation buffer. In this case, the BGE has a high pH thermal coefficient whereas the carboxylic groups of the analytes have low pK(a) thermal coefficients. The application of these dynamic thermal gradients increased peak height by a factor of two (and decreased the standard deviations of peaks by a factor of two) of aspartic acid and glutamic acid derivatized with naphthalene-2,3-dicarboxaldehyde and serine derivatized with fluorescamine. The effect of thermal compression of bands was not observed when runs were accomplished using phosphate buffer at pH 7 (negative control). Phosphate has a low dpH/dT in this pH range, similar to the dK(a)/dT of analytes. It is shown that vertical bar dK(a)/dT-dpH/dT vertical bar >> 0 is one determinant factor to have significant stacking produced by dynamic thermal junctions.

Sample stacking in CZE using dynamic thermal junctions II: Analytes with high dpK(a)/dT crossing a single thermal junction in a BGE with low dpH/dT

In a previous work [M. Mandaji, et al., this issue] a sample stacking method was theoretically modeled and experimentally demonstrated for analytes with low dpK(a)/dT (analytes carrying carboxylic groups) and BGEs with high dpH/dT (high pH-temperature-coefficients). In that work, buffer pH was modulated with temperature, inducing electrophoretic mobility changes in the analytes. In the present work, the opposite conditions are studied and tested, i.e. analytes with high dpK(a)/dT and BGEs that exhibit low dpH/dT. It is well known that organic bases such as amines, imidazoles, and benzimidazoles exhibit high dpK(a)/dT. Temperature variations induce instantaneous changes on the basicity of these and other basic groups. Therefore, the electrophoretic velocity of some analytes changes abruptly when temperature variations are applied along the capillary. This is true only if BGE pH remains constant or if it changes in the opposite direction of pK(a) of the analyte. The presence of hot and cold sections along the capillary also affects local viscosity, conductivity, and electric field strength. The effect of these variables on electrophoretic velocity and band stacking efficacy was also taken into account in the theoretical model presented. Finally, this stacking method is demonstrated for lysine partially derivatized with naphthalene-2,3-dicarboxaldehyde. In this case, the amino group of the lateral chain was left underivatized and only the alpha amino group was derivatized. Therefore, the basicity of the lateral amino group, and consequently the electrophoretic mobility, was modulated with temperature while the pH of the buffer used remained unchanged.

On the molecular mass of the extracellular hemoglobin of Glossoscolex paulistus: Analytical ultracentrifugation reexamination

The giant extracellular hemoglobin of Glossoscolex paulistus (HbGp) is constituted by Subunits containing heme groups with molecular masses (M) in the range of 15 to 19 kDa, monomers of 16 kDa (d), and trimers of 51 to 52 kDa (abc) linked by nonheme structures named linkers of 24 to 32 kDa (L). HbGp is homologous to Lumbricus terrestris hemoglobin (HbLt). Several reports propose M of HbLt in the range of 3.6 to 4.4 MDa. Based on subunits M determined by mass spectrometry and assuming HbGp stoichiometry of 12(abcd)(3)L(3) (Vinogradov model) plus 144 heme groups, a Value of M for HbGp oligomer of 3560 kDa can be predicted. This Value is nearly 500 kDa higher than the unique HbGp M Value reported in the literature. In the current work, sedimentation velocity analytical ultracentrifugation (AUC) experiments were performed to obtain M for HbGp in oxy and cyano-met forms. s(20,w)(0), values of 58.1 +/- 0.2 S and 59.6 +/- 0.2 S, respectively, for the two oxidation forms were obtained. The ratio between sedimentation and diffusion coefficients supplied values for M of approximately 3600 100 and 3700 100 kDa for oxy and cyano-met HbGp forms, respectively. An independent determination of the partial specific volume, V(bar), for HbGp was performed based on density measurements, providing a value of 0.764 +/- 0.008, in excellent agreement with the estimates from SEDFIT software. Our results show total consistency between M obtained by AUC and recent partial characterization by mass spectrometry. Therefore, HbGp possesses M very close to that of HbLt, suggesting an oligomeric assembly in agreement with the Vinogradov model. (c) 2008 Elsevier Inc. All rights reserved.

Analytical Assessment of a Home Made Capillary Electrophoresis Equipment with Linear Charge Coupled Device for Visible Light Absorption Detection in the Determination of Food Dyes

The performance of modular home made capillary electrophoresis equipment with spectrophotometric detection, at a visible region by means of a miniaturized linear charge coupled device, was evaluated for the determination of four food dyes. This system presents a simple but efficient home made cell detection scheme. A computer program that converts the spectral data after each run into the electropherograms was developed to evaluate the analytical parameters. The dyes selected for analytical evaluation of the system were Brilliant Blue FCF, Fast Green FCF, Sunset Yellow FCF, and Amaranth. Separation was carried out in a 29cm length and 75 mu m I.D fused silica capillary, using 10mmolL-1 borate buffer at pH 9, with separation voltage of 7.5kV. The detection limits for the dyes were between 0.3 and 1.5mgL-1 and the method presented adequate linearity over the ranges studied, with correlation coefficients greater than 0.99. The method was applied for determination and quantification of these dyes in fruit juices and candies.

TLC Separation and Identification of the Essential Oil Constituents from Aloysia gratissima

The present work had as objective the isolation of the five compounds by thin-layer Chromatography (TLC) from the essential oil of the Aloysia gratissima. For this, a number of systems of eluents were used for its separation, indicating that through the system acetone/hexane in proportions (v/v) 1:30 it was possible to isolate guaiol, elemol, pinocanphone (trans-3-pinanone), cis-pinocarvyl, and acorenone. The isolation of the compound acorenone from the other compounds was possible with the mixture of solvents hexane/dichloromethane in proportions (v/v) (1:1,3).

Comparison of the analytical performance of electrophoresis microchannels fabricated in PDMS, glass, and polyester-toner

This paper compares the analytical performance of microchannels fabricated in PDMS, glass, and polyester-toner for electrophoretic separations. Glass and PDMS chips were fabricated using well-established photolithographic and replica-molding procedures, respectively. PDMS channels were sealed against three different types of materials: native PDMS, plasma-oxidized PDMS, and glass. Polyester-toner chips were micromachined by a direct-printing process using an office laser printer. All microchannels were fabricated with similar dimensions according to the limitations of the direct-printing process (width/depth 150 mu m/12 mu m). LIF was employed for detection to rule out any losses in separation efficiency due to the detector configuration. Two fluorescent dyes, coumarin and fluorescein, were used as model analytes. Devices were evaluated for the following parameters related to electrophoretic separations: EOF, heat dissipation, injection reproducibility, separation efficiency, and adsorption to channel wall.

ANALYSIS & STUDY OF AI TECHNIQUES FORAUTOMATIC CONDITION MONITORING OFRAILWAY TRACK INFRASTRUCTURE : Artificial Intelligence Techniques

Since the last decade the problem of surface inspection has been receiving great attention from the scientific community, the quality control and the maintenance of products are key points in several industrial applications.The railway associations spent much money to check the railway infrastructure. The railway infrastructure is a particular field in which the periodical surface inspection can help the operator to prevent critical situations. The maintenance and monitoring of this infrastructure is an important aspect for railway association.That is why the surface inspection of railway also makes importance to the railroad authority to investigate track components, identify problems and finding out the way that how to solve these problems. In railway industry, usually the problems find in railway sleepers, overhead, fastener, rail head, switching and crossing and in ballast section as well. In this thesis work, I have reviewed some research papers based on AI techniques together with NDT techniques which are able to collect data from the test object without making any damage. The research works which I have reviewed and demonstrated that by adopting the AI based system, it is almost possible to solve all the problems and this system is very much reliable and efficient for diagnose problems of this transportation domain. I have reviewed solutions provided by different companies based on AI techniques, their products and reviewed some white papers provided by some of those companies. AI based techniques likemachine vision, stereo vision, laser based techniques and neural network are used in most cases to solve the problems which are performed by the railway engineers.The problems in railway handled by the AI based techniques performed by NDT approach which is a very broad, interdisciplinary field that plays a critical role in assuring that structural components and systems perform their function in a reliable and cost effective fashion. The NDT approach ensures the uniformity, quality and serviceability of materials without causing any damage of that materials is being tested. This testing methods use some way to test product like, Visual and Optical testing, Radiography, Magnetic particle testing, Ultrasonic testing, Penetrate testing, electro mechanic testing and acoustic emission testing etc. The inspection procedure has done periodically because of better maintenance. This inspection procedure done by the railway engineers manually with the aid of AI based techniques.The main idea of thesis work is to demonstrate how the problems can be reduced of thistransportation area based on the works done by different researchers and companies. And I have also provided some ideas and comments according to those works and trying to provide some proposal to use better inspection method where it is needed.The scope of this thesis work is automatic interpretation of data from NDT, with the goal of detecting flaws accurately and efficiently. AI techniques such as neural networks, machine vision, knowledge-based systems and fuzzy logic were applied to a wide spectrum of problems in this area. Another scope is to provide an insight into possible research methods concerning railway sleeper, fastener, ballast and overhead inspection by automatic interpretation of data.In this thesis work, I have discussed about problems which are arise in railway sleepers,fastener, and overhead and ballasted track. For this reason I have reviewed some research papers related with these areas and demonstrated how their systems works and the results of those systems. After all the demonstrations were taking place of the advantages of using AI techniques in contrast with those manual systems exist previously.This work aims to summarize the findings of a large number of research papers deploying artificial intelligence (AI) techniques for the automatic interpretation of data from nondestructive testing (NDT). Problems in rail transport domain are mainly discussed in this work. The overall work of this paper goes to the inspection of railway sleepers, fastener, ballast and overhead.

Pregnant Adolescents in Vietnam : Social context and health care needs

Background: The number of childbearing adolescents in Vietnam is relatively low but they are more prone to experience adverse outcome than adult women. Reports of increasing rates of abortion and prevalence of STIs including HIV among youth indicate a need to improve services and counselling for these groups. Midwives are key persons in the promotion of young people’s sexual and reproductive health in Vietnam. Aim: The overall aim of this thesis is to describe the prevalence and outcome of adolescent pregnancies in Vietnam (I), to explore the social context and health care seeking behavior of pregnant adolescents (II), as well as to explore the perspectives of health care providers and midwifery students regarding adolescent sexuality and reproductive health service needs (III, IV). Methods: The studies were conducted from 2002 to 2005, combining qualitative and quantitative research methods. A population based prospective survey was used to estimate rates and outcomes of adolescent pregnancies (I). Pregnant and newly delivered adolescents’ experiences of childbearing and their encounters with health care providers were studied using qualitative interviews (II). Health care providers’ perspective on adolescent sexual and reproductive health (ASRH) and views on how to improve the quality of abortion care was explored in focus group discussions (FGD). The values and attitudes of midwifery students about ASRH were investigated using questionnaires and interviews (IV). Descriptive statistics was used to analyse quantitative data (I, IV) and content analysis were applied for qualitative data (II, III, and IV). Findings: Adolescent birth rate was similar to previously reported in Vietnam but lower when compared to other Asian countries. The incidence of stillborn among adolescents was higher than for women in higher reproductive ages. The proportion of preterm deliveries was 20 % of all births, higher than previous findings from Vietnam. About 2 % of the deliveries were home deliveries, more common among women with low education, belonging to ethnic minority and/or living in mountainous areas (I). Ambivalence facing motherhood, pride and happiness but also worries and lack of self-confidence emerged as themes from the interviews; and experience of ‘being in the hands of others’ in a positive, caring sense but also in a sense of subordination in relation to husband, family and health care providers (II). Health care providers at abortion clinics and midwifery students generally disapproved of pre-marital sex, but had a pragmatic view on the need for contraceptive services and counselling to reduce the burden of unwanted pregnancies and abortions for young women. Providers and midwifery students expressed a need for training on ASRH issues (III, IV). Conclusion: Cultural norms and gender inequity make pregnant adolescent women in Vietnam vulnerable to sexual and reproductive health risks. Health care providers experience ethical dilemmas while counselling unmarried adolescents who come for abortion and this has a negative impact on the quality of care. Integrated ASRH in education and training programmes for health care providers, including midwives, as well as continued in-service training on these issues are suggested to improve reproductive health care services in Vietnam.

Review of the literature on credit risk modeling : development of the past 10 years

This paper traces the developments of credit risk modeling in the past 10 years. Our work can be divided into two parts: selecting articles and summarizing results. On the one hand, by constructing an ordered logit model on historical Journal of Economic Literature (JEL) codes of articles about credit risk modeling, we sort out articles which are the most related to our topic. The result indicates that the JEL codes have become the standard to classify researches in credit risk modeling. On the other hand, comparing with the classical review Altman and Saunders(1998), we observe some important changes of research methods of credit risk. The main finding is that current focuses on credit risk modeling have moved from static individual-level models to dynamic portfolio models.

Non-challenging education and teacher control as factors for marginalization of students in diverse settings

This article discusses teachers’ attitudes towards immigrant students in poor settings and the effect these attitudes have on organization of education on classroom level. It draws on results from two ethnographic studies where some primary school classes in Sweden were followed with participant observation and interviews as main research methods. The article focuses on classroom activities and teachers’ attitudes towards immigrant students and students with low socio-economic status. In the article is argued for the importance of presenting students in poor settings with demanding tasks and challenging education. In these cases, intellectually undemanding tasks in combination with little room for students’ own initiatives resulted in low enthusiasm among students regarding schoolwork and accordingly low learning, while classroom work that demanded active involvement by students in combination with high level of students’ influence on what took place in classrooms resulted in high level of students’ engagement and high outcome.

Degree projects improving co-operation between the teacher education program and schools : - Swedish experiences

In 1977, when teacher education inSwedenwas incorporated into the university system, the main reason was to transform it into an academic tradition. Now, nearly 30 years later, there is still tension between the academic and the vocational tradition; they show up as different and separated elements in a program that is meant to prepare students for a career as a teacher and for a possible future career as a researcher in this field. This tension gives rise to a risk of allowing parallel “tracks” to develop and of isolating the degree thesis work from other courses. On the teacher-training program in which we are involved, close co-operation with partner-schools, where the practical part of the program takes place, has been established. Here, the students´ degree theses are an important factor in making this co-operation work on a concrete level. Accordingly, the purpose of the degree thesis is both to reflect problems in schools and contribute to better teaching in the individual school, but also to offer relevant educational subject matter to the students, which may be adapted as an element in the university’s research environments. From these points of view, the degree thesis is an important part of teacher education and rather more than just a single course. The degree thesis should be an element that gives the students an opportunity to show that they have reached central goals in the teacher education program. It should also be an integral part in the development of critical and scholarly thinking, deepening pedagogical and didactic knowledge and giving the students an opportunity to apply research methods. We will here use two minor case studies: one that compares teacher education programs in five Swedish universities and one minor study at one of those universities, in order to elaborate on the questions: - What purpose has a degree thesis and what role does it play in teacher education? - What criteria are relevant to assessing a degree thesis and what qualities do the degree theses have? - Is it possible to assess a degree thesis fairly and what happens to students who do not pass? - How could the degree thesis be used to improve the contact between the teacher education program and its partner schools in order to contribute to the development of the individual school?

Desenvolvimento de um modelo de intervenção baseado no sistema toyota de produção e na teoria das restrições : autilização da pesquisa-ação em uma indústria de cerâmica vermelha de pequeno porte da região metropolitana de Porto Alegre

As teorias de gestão da produção, como o Sistema Toyota de Produção e a Teoria das Restrições, têm apresentado resultados positivos, em realidades organizacionais muito diferenciadas. Contudo, é preciso garantir a efetividade das ações em provocar as mudanças desejadas. Neste sentido, métodos de pesquisa participativa, como a pesquisaação, promovem a participação e o comprometimento das pessoas implicadas no processo de mudança. Esta dissertação propõe a construção de um modelo de intervenção visando aumentar a competitividade de uma realidade organizacional específica. Foram utilizados a pesquisa-ação, como método de trabalho e a Teoria das Restrições (TOC) e o Sistema Toyota de Produção (STP), como embasamento teórico. Cabe ressaltar que este modelo foi construído a partir de uma intervenção realizada em uma indústria de cerâmica vermelha da região metropolitana de Porto Alegre. Assim, a presente dissertação foi organizada da seguinte maneira: revisão bibliográfica do método de condução da pesquisa e adaptação do mesmo para o presente trabalho, fundamentação teórica, composta pelos princípios básicos de sustentação do STP e da TOC, análise do contexto do segmento industrial em questão, descrição da intervenção realizada e apresentação do modelo construído, análise dos resultados finais, conclusões e recomendações para futuras pesquisas. A análise dos resultados obtidos e as conclusões do estudo revelam a possibilidade de generalização parcial do modelo proposto, desde que observadas as características específicas da realidade industrial em questão.

Rádio peão: estudo dos mecanismos, tipologia e os efeitos dos rumores nas organizações

Esta dissertação tem como tema o estudo da rádio peão, equivalente brasileiro da grapevine americana e dos rumores e boatos que circulam nas organizações. O trabalho contém a revisão da teoria existente sobre o assunto, abrangendo seus principais mecanismos de . surgimento e transformação, a tipologia, e classificação dos rumores, segundo seus aspectos sociais e psicológicos. Também analisa as diversas metodologias de pesquisa do assunto. Ao final são feitas algumas recomendações para as empresas e sugeridos alguns temas para aprofundamento futuro do assunto.

Governança e coordenação em arranjos multinível de políticas públicas transversais: assistência social, educação e saúde no Programa Bolsa Família

Este estudo apresenta o desenvolvimento teórico e a aplicação de um modelo voltado à análise de arranjos de governança de políticas públicas que ocorrem quando múltiplas organizações estão envolvidas em lidar com questões complexas. A seguinte pergunta de pesquisa foi proposta: Como opera o conjunto de instrumentos de coordenação empregados na governança de uma política pública transversal? Um estudo de caso exploratório sobre o Programa Bolsa Família foi realizado, especificamente na sua dimensão relacionada à coordenação entre os setores de assistência social, educação e saúde. Os métodos de pesquisa consistiram em análise de conteúdo de documentos e transcrições de entrevistas semiestruturadas com atores-chave do Programa nos três setores de políticas. Os resultados demonstram como diferentes modos de governança coexistem no arranjo multinível estudado, revelando diversos elementos de coordenação cuja operação concomitante resulta em um modo de governança misto. Contribui-se, assim, para o aperfeiçoamento do Programa e para o desenvolvimento teórico, metodológico e aplicado dos conceitos de governança e de coordenação, ficando demonstrado ser possível integrá-los na análise de políticas públicas transversais.