Tailoring Heme-Thiolate Proteins into Efficient Biocatalysts with High Specificity and Selectivity

Cytochrome P450 monooxygenases, one of the most important classes of heme-thiolate proteins, have attracted considerable interest in the biochemical community because of its catalytic versatility, substrate diversity and great number in the superfamily. Although P450s are capable of catalyzing numerous difficult oxidation reactions, the relatively low stability, low turnover rates and the need of electron-donating cofactors have limited their practical biotechnological and pharmaceutical applications as isolated enzymes. The goal of this study is to tailor such heme-thiolate proteins into efficient biocatalysts with high specificity and selectivity by protein engineering and to better understand the structure-function relationship in cytochromes P450. In the effort to engineer P450cam, the prototype member of the P450 superfamily, into an efficient peroxygenase that utilizes hydrogen peroxide via the “peroxide-shunt” pathway, site-directed mutagenesis has been used to elucidate the critical roles of hydrophobic residues in the active site. Various biophysical, biochemical and spectroscopic techniques have been utilized to investigate the wild-type and mutant proteins. Three important P450cam variants were obtained showing distinct structural and functional features. In P450camV247H mutant, which exhibited almost identical spectral properties with the wild-type, it is demonstrated that a single amino acid switch turned the monooxygenase into an efficient preoxidase by increasing the peroxidase activity nearly one thousand folds. In order to tune the distal pocket of P450cam with polar residues, Leu 246 was replaced with a basic residue, lysine, resulting in a mutant with spectral features identical to P420, the inactive species of P450. But this inactive-species-like mutant showed catalytic activities without the facilitation of any cofactors. By substituting Gly 248 with a histidine, a novel Cys-Fe-His ligation set was obtained in P450cam which represented the very rare case of His ligation in heme-thiolate proteins. In addition to serving as a convenient model for hemoprotein structural studies, the G248H mutant also provided evidence about the nature of the axial ligand in cytochrome P420 and other engineered hemoproteins with thiolate ligations. Furthermore, attempts have been made to replace the proximal ligand in sperm whale myoglobin to construct a heme-thiolate protein model by mimicking the protein environment of cytochrome P450cam and chloroperoxidase.

Decisões comportamentais em contexto de forrageamento de dinoponera quadriceps

The behavioral decisions of animals do not occur randomly, because behaviors are adjusted to ensure the survival and reproduction of the animal. In this research, I examined behavioral decisions in the foraging context of the ant Dinoponera quadriceps with regard to orientation, food avaliation and foraging dynamic to individual level. The study was conducted at the Laboratory of Behavioral Biology at UFRN and in an area of secondary Atlantic Forest in FLONA-ICMBio Nísia Floresta/RN. In all observations and experiments, ants were marked individually with an alphanumeric code label fixed on the thorax. In the first part of the study, I analyzed the orientation cues used by D. quadriceps. The tests were performed in a maze of 17 compartments. Each forager was tested for 10 min in three sessions for six different treatments. The treatments consisted of the presence or absence of odor and superior or frontal visual cues. The workers demonstrated that the presence of odor is indispensable and front visual cues are more effective than superior visual cues. In the second part, I investigated the discrimination of food, considering the parameters, size, weight and volume. In a 'cafeteria' experiment, I offered cylindrical pieces of food (mortadella) in a Petri dish, within an experimental arena 1m². Initially, the pieces were of four different sizes; in a second step, the pieces were of the same size but with different weight; in the last step, the pieces had the same weight but different volumes. The results showed the effect of the size and weight parameters for food choice. In the third part of the study, I evaluated the influence of the activity of active foragers on inactive ones. In this part, the colonies were observed in a natural environment. The observations took place on three consecutive days in 10 episodes, total of 30 days for each colony, 12 hours/day. On the first day, I registered the output and input of workers; on the second day, the most active ants on the first day were taken and given back at the end of the observations; on the third day, the observations were similar to the first day. As a result, the workers of D. quadriceps show autostimulation and they do not show social facilitation and the colony compensates the absence of the most active workers. Based on the stated, I conclude that workers of D. quadriceps use chemical, frontal and superior visual orientation cues during their displacements. They discriminate the chosen food by size and weight. The regulation of activity dynamics of foragers is by autostimulation, an active worker does not influence the activity of an inactive worker, the successful search previous is the stimulus to the successful worker itself to continue foraging activity.

Semantikos: proposta de interfaces para aplicativo mediador de técnica de diferencial semântico em mobile tablet

This research aims to systematize a proposal of developing a mobile tablet application in order to help implementing the Semantic Differential technique – SD, under the approach of Participatory Design. In 1975, Osgood et al. created the Semantic Differential technique. Since then, many experiments use it to measure the affective perception of individuals concerning objects and concepts by means of compounded scales of bipolar adjectives, based on the theoretical models that support the technique: the conductible, spatial and metric models. During the application of the technique with potential users, the researcher must simultaneously manage several contexts, that is, audio recorder, when authorized, and observe and record spontaneous reports of the respondent. It is noticeable that often occurs a cognitive overload during this event. Thus, the use of a single application whose interface is assigned to its users and respondents could assist researchers in applying the SD technique. This research aimed to understand the processes inherent to the task of implementing the Semantic Differential technique and obeyed the following steps: a) training of users, b) background questionnaire c) interview with Focus Group, and d) cooperative evaluation. Besides these procedures, one can also observe the degrees of facilitation or difficulty concerning the use of the conventional model, which is the development and application of scales with the aid of printed material, pencil, pens, clipboards, and recorder software for editing the document and data analysis. This paper comprises reactions and impressions from the experiences of users of SD technique. Considering the data recollected from the user’s observation, the hypothesis of the experiment proved to be right. It means that the development of the application for mobile tablet employing the technique of Semantic Differential is viable, since it assembles all the steps in one only tool, increases the accomplishment of the task between user/researcher and user/respondent resulting in their mutual satisfaction.

Plant diversity influencing structure and functioning of Caatinga vegetation

O estudo dos efeitos que a diversidade de espécies pode causar nos processos ecossistêmicos tem crescido vertiginosamente nas últimas duas décadas. Diversos trabalhos experimentais realizados no mundo todo têm demonstrado que uma maior diversidade de plantas contribui para o aumento da produtividade de ecossistemas terrestres. Além disso, esse efeito pode influenciar processos em diversos níveis tróficos, contribuindo assim para a estabilidade dos processos ecossistêmicos a longo prazo. Paralelamente com os estudos do efeito da diversidade, muita atenção tem sido dada para desvendar o papel das características funcionais das espécies no funcionamento dos ecossistemas. Isto porque as características funcionais das espécies têm se mostrado importantes "peças" no entendimento dos efeitos que espécies individuais podem exercer nos ecossistemas e suas respostas ao ambiente. Nesta tese de doutorado eu explorei algumas lacunas de conhecimento dentro dessa área em crescente desenvolvimento conhecida na literatura ecológica como "biodiversidade e funcionamento dos ecossistemas". No primeiro capítulo, eu busquei evidências para mecanismos que podem explicar a relação positiva entre diversidade e funcionamento com foco em cinco mecanismos relacionados às interações entre plantas, tendo como parâmetro de funcionamento a produtividade primária. No segundo capítulo, eu utilizei técnicas para a estimativa de padrões de diversidade em escalas biogeográficas e bases de dados de satélites com longa duração para desvendar se a biodiversidade em escalas macroecológicas promove a estabilidade da produtividade dos ambientes terrestres no semiárido brasileiro. Por fim, o objetivo do terceiro capítulo foi entender como a perda da cobertura vegetal originária do uso da terra por comunidades tradicionais no semiárido brasileiro influenciaria os processos de interações entre plantas e o papel das características funcionais das espécies nessas interações. Acredito que a contribuição individual de cada capítulo preenche lacunas de conhecimento importantes dessa área da Ecologia que ainda se encontra em expansão.

Relational restorative justice pedagogy in educator professional development

What would a professional development experience rooted in the philosophy, principles, and practices of restorative justice look and feel like? This article describes how such a professional development project was designed to implement restorative justice principles and practices into schools in a proactive, relational and sustainable manner by using a comprehensive dialogic, democratic peacebuilding pedagogy. The initiative embodied a broad, transformative approach to restorative justice, grounded in participating educators’ identifying, articulating and applying personal core values. This professional development focused on diverse educators, their relationships, and conceptual understandings, rather than on narrow techniques for enhancing student understanding or changing student behaviour. Its core practice involved facilitated critical reflexive dialogue in a circle, organized around recognizing the impact of participants’ interactions on others, using three central, recurring questions: Am I honouring? Am I measuring? What message am I sending? Situated in the context of relational theory (Llewellyn, 2012), this restorative professional development approach addresses some of the challenges in implementing and sustaining transformative citizenship and peacebuilding pedagogies in schools. A pedagogical portrait of the rationale, design, and facilitation experience illustrates the theories, practices, and insights of the initiative, called Relationships First: Implementing Restorative Justice From the Ground Up.

Sexualidade e saúde mental: construindo sentidos entre pessoas usuárias de um CAPS

When expressed by mental health services users, sexuality is typically denied by professionals, viewed as another symptom or as if these people are not capable of practicing it. Once Brazilian health professionals haven’t shown lots of investment in this theme, and few are the studies in this field, it is necessary the attention to be focused on researches involving this public. Therefore, the main goal of this study was understand the meanings of sexuality of the mental health services users, which were negotiated in sexuality workshops. The secondary goals were: a) understand the meanings of themes about sexuality brought by users through their experiences of everyday life; b) to evaluate the facilitating experience of the workshops on sexuality at CAPS. Thus, 10 workshops on sexuality were held, with an average of an hour and twenty minutes each, distributed from December 2014 and April 2015. There were 43 participants, 29 women and 14 men. The meetings had the following central themes: sexuality; sexuality and mental health; myths, beliefs and sexual taboos; gender identity; sexual orientation; sexual and reproductive rights; safe sex; and STD/AIDS. The data collection was through audio-recording of these meetings. Later, was made the transcript of the workshops, a careful reading of these transcripts and then its analysis. It was identified categories to analyze the interfaces that permeate the focus of the study. Initially, the categories relating to mental health and sexuality: meanings about sexuality; gender issues; gender and religion; sexual rights, STD/AIDS prevention and attention or denial of sexuality at CAPS. Later, those relating to the workshops facilitating process: challenges in facilitating the workshops; and the perception of the participants. A variety of meanings about sexuality could be noticed in the users’ statements, relating it more with affection and respect than with intercourse. The gender issues that emerged during the workshops were related to marital relationship, sexism, domestic violence, psychological violence and male and female roles in society. Moreover, were also revealed some situations that associated gender differences with religious issues, such as the submission of women and homosexuality. It was also noticed some experiences of the participants involving worrying situations of family violence, suicidal ideation and chemical castration, were often mismanaged or ignored by the service professionals. With regard to the facilitation of the workshops, it was possible to legitimize it as places where users were able to talk openly about the suggested themes and highlight its importance to the study site. Besides, it’s possible to list a few challenges of its facilitation in a mental health service, which was in general positively evaluated by the participants. Thus, the research highlights the need for sexuality theme discussion in mental health services, in order to understand, discuss and inform the users. Also, it’s important to problematize the stigma created in the theme relation with the users, the professionals and the society, working its specificities and avoiding a pathological bias.

Psychometric functions for detection and discrimination with and without flankers.

Recent studies have reported that flanking stimuli broaden the psychometric function and lower detection thresholds. In the present study, we measured psychometric functions for detection and discrimination with and without flankers to investigate whether these effects occur throughout the contrast continuum. Our results confirm that lower detection thresholds with flankers are accompanied by broader psychometric functions. Psychometric functions for discrimination reveal that discrimination thresholds with and without flankers are similar across standard levels, and that the broadening of psychometric functions with flankers disappears as standard contrast increases, to the point that psychometric functions at high standard levels are virtually identical with or without flankers. Threshold-versus-contrast (TvC) curves with flankers only differ from TvC curves without flankers in occasional shallower dippers and lower branches on the left of the dipper, but they run virtually superimposed at high standard levels. We discuss differences between our results and other results in the literature, and how they are likely attributed to the differential vulnerability of alternative psychophysical procedures to the effects of presentation order. We show that different models of flanker facilitation can fit the data equally well, which stresses that succeeding at fitting a model does not validate it in any sense.

Motivational, volitional and multiple goal predictors of walking in people with type 2 diabetes

Acknowledgment MN's PhD scholarship was provided by Ministry of Health and Medical Education (Islamic Republic of Iran). This study was funded by the University of Aberdeen. FFS is funded by Fuse, the UK Clinical Research Collaboration Centre of Excellence for Translational Research in Public Health. The researchers gratefully acknowledge all the Type 2 diabetic patients and their household members who participated in the study for their contribution to this study; without them there would be no data. The researchers gratefully acknowledge the SDRN for providing the list of Type 2 diabetes and helping for sampling.

Motivational, volitional and multiple goal predictors of walking in people with type 2 diabetes

Acknowledgment MN's PhD scholarship was provided by Ministry of Health and Medical Education (Islamic Republic of Iran). This study was funded by the University of Aberdeen. FFS is funded by Fuse, the UK Clinical Research Collaboration Centre of Excellence for Translational Research in Public Health. The researchers gratefully acknowledge all the Type 2 diabetic patients and their household members who participated in the study for their contribution to this study; without them there would be no data. The researchers gratefully acknowledge the SDRN for providing the list of Type 2 diabetes and helping for sampling.

Les valves sans suture dans le traitement de la sténose aortique sévère

Rapport présenté en vue de l’obtention du grade de M.Sc.A génie biomédical option génie clinique

Analysis of Global Compliance and Implementation of the Goals of International Environmental Treaties: A Case study of the Convention on Biodiversity (CBD)

The Convention on Biodiversity (CBD) was created in 1992 to coordinate global governments to protect biological resources. The CBD has three goals: protection of biodiversity, achievement of sustainable use of biodiversity and facilitation of equitable sharing of the benefits of biological resources. The goal of protecting biological resources has remained both controversial and difficult to implement. This study focused more on the goal of biodiversity protection. The research was designed to examine how globally constructed environmental policies get adapted by national governments and then passed down to local levels where actual implementation takes place. Effectiveness of such policies depends on the extent of actual implementation at local levels. Therefore, compliance was divided and examined at three levels: global, national and local. The study then developed various criteria to measure compliance at these levels. Both qualitative and quantitative methods were used to analyze compliance and implementation. The study was guided by three questions broadly examining critical factors that most influence the implementation of biodiversity protection policies at the global, national and local levels. Findings show that despite an overall biodiversity deficit of 0.9 hectares per person, global compliance with the CBD goals is currently at 35%. Compliance is lowest at local levels at 14%, it is slightly better at national level at 50%, and much better at the international level 64%. Compliance appears higher at both national and international levels because compliance here is paper work based and policy formulation. If implementation at local levels continues to produce this low compliance, overall conservation outcomes can only get worse than what it is at present. There are numerous weaknesses and capacity challenges countries are yet to address in their plans. In order to increase local level compliance, the study recommends a set of robust policies that build local capacity, incentivize local resource owners, and implement biodiversity protection programs that are akin to local needs and aspirations.

Burrowing times and proportions of Mesodesma donacium

Data archives with raw data of burrowing times and proportions of succesfully burrowed clams.

Facilitating Inclusion: Austrian Wisdom Councils as Democratic Innovation between Consensus and Diversity

Democratic innovations face the challenge of realizing deliberative democratic ideals in the context of structural inequality. Consensus decision making and expertise have been said to have exclusive effects on marginalized groups like women and ethnic and sexual minorities, which obstructs diversity. Wisdom Councils as practiced in Austria attempt to counter inequalities by including marginalized groups through the moderation technique dynamic facilitation. Exploratory participatory observations and interviews with a moderator and the participants of two Wisdom Councils in Austria provide a deeper understanding of the inclusive processes at work in Wisdom Councils facilitating a productive combination of consensus and diversity.

Juniperus navicularis - a study of Portuguese population structure and fitness

Juniperus navicularis Gand. is a dioecious endemic conifer that constitutes the understory of seaside pine forests in Portugal, areas currently threatened by increasing urban expansion. The aim of this study is to assess the conservation status of previously known populations of this species located on its core area of distribution. The study was performed in south-west coast of Portugal. Three populations varying in size and pine density were analyzed. Number of individuals, population density, spatial distribution and individual characteristics of junipers were estimated. Female cone, seed characteristics and seed viability were also evaluated. Results suggest that J. navicularis populations are vulnerable because seminal recruitment is scarce, what may lead to a reduction of genetic variability due solely to vegetative propagation. This vulnerability seems to be strongly determined by climatic constraints toward increasing aridity. Ratio between male and female shrubs did not differ from 1:1 in any population. Deviations from 1:1 between mature and non-mature plants were found in all populations, denoting population ageing. Very low seed viability was observed. A major part of described Juniperus navicularis populations have disappeared through direct habitat loss to urban development, loss of fitness in drier and warmer locations and low seed viability. This study is the first to address J. navicularis conservation, and represents a valuable first step toward this species preservation. 

La música en el desarrollo de la espiritualidad y la religiosidad. Una aproximación al Cristianismo y al Budismo

La música puede afectar al individuo en todos sus niveles –físico, mental y espiritual–. El presente artículo se centra en el papel que ésta desempeña en el desarrollo de la vida espiritual y trascendental. Para ello, realizaremos un repaso histórico de su evolución estética y social, abordaremos dicho fenómeno a nivel fisiológico y presentaremos sus aplicaciones clínicas y sociales. Seguidamente y a modo de ejemplo de las concepciones de pensamiento occidental y oriental, trataremos la forma en que el cristianismo y el budismo conciben la música dentro de su doctrina. Finalizaremos con algunas reflexiones sobre el tema.