Glutathione Transferases as Detoxification Agents: Structural and Functional Studies

Glutathione transferases (GSTs) are a diverse family of enzymes that catalyze the glutathione-dependent detoxification of toxic compounds. GSTs are responsible for the conjugation of the tripeptide glutathione (GSH) to a wide range of electrophilic substrates. These include industrial pollutants, drugs, genotoxic carcinogen metabolites, antibiotics, insecticides and herbicides. In light of applications in biomedicine and biotechnology as cellular detoxification agents, detailed structural and functional studies of GSTs are required. Plant tau class GSTs play crucial catalytic and non-catalytic roles in cellular xenobiotic detoxification process in agronomically important crops. The abundant existence of GSTs in Glycine max and their ability to provide resistance to abiotic and biotic stresses such as herbicide tolerance is of great interest in agriculture because they provide effective and suitable tools for selective weed control. Structural and catalytic studies on tau class GST isoenzymes from Glycine max (GmGSTU10-10, GmGSTU chimeric clone 14 (Sh14), and GmGSTU2-2) were performed. Crystal structures of GmGSTU10-10 in complex with glutathione sulfenic acid (GSOH) and Sh14 in complex with S-(p-nitrobenzyl)-glutathione (Nb-GSH) were determined by molecular replacement at 1.6 Å and 1.75 Å, respectively. Major structural variations that affect substrate recognition and catalytic mechanism were revealed in the upper part of helix H4 and helix H9 of GmGSTU10-10. Structural analysis of Sh14 showed that the Trp114Cys point mutation is responsible for the enhanced catalytic activity of the enzyme. Furthermore, two salt bridges that trigger an allosteric effect between the H-sites were identified at the dimer interface between Glu66 and Lys104. The 3D structure of GmGSTU2-2 was predicted using homology modeling. Structural and phylogenetic analysis suggested GmGSTU2-2 shares residues that are crucial for the catalytic activity of other tau class GSTs–Phe10, Trp11, Ser13, Arg20, Tyr30, Leu37, Lys40, Lys53, Ile54, Glu66 and Ser67. This indicates that the catalytic and ligand binding site in GmGSTU2-2 are well-conserved. Nevertheless, at the ligandin binding site a significant variation was observed. Tyr32 is replaced by Ser32 in GmGSTU2-2 and thismay affect the ligand recognition and binding properties of GmGSTU2-2. Moreover, docking studies revealed important amino acid residues in the hydrophobic binding site that can affect the substrate specificity of the enzyme. Phe10, Pro12, Phe15, Leu37, Phe107, Trp114, Trp163, Phe208, Ile212, and Phe216 could form the hydrophobic ligand binding site and bind fluorodifen. Additionally, side chains of Arg111 and Lys215 could stabilize the binding through hydrogen bonds with the –NO2 groups of fluorodifen. GST gene family from the pathogenic soil bacterium Agrobacterium tumefaciens C58 was characterized and eight GST-like proteins in A. tumefaciens (AtuGSTs) were identified. Phylogenetic analysis revealed that four members of AtuGSTs belong to a previously recognized bacterial beta GST class and one member to theta class. Nevertheless, three AtuGSTs do not belong to any previously known GST classes. The 3D structures of AtuGSTs were predicted using homology modeling. Comparative structural and sequence analysis of the AtuGSTs showed local sequence and structural characteristics between different GST isoenzymes and classes. Interactions at the G-site are conserved, however, significant variations were seen at the active site and the H5b helix at the C-terminal domain. H5b contributes to the formation of the hydrophobic ligand binding site and is responsible for recognition of the electrophilic moiety of the xenobiotic. It is noted that the position of H5b varies among models, thus providing different specificities. Moreover, AtuGSTs appear to form functional dimers through diverse modes. AtuGST1, AtuGST3, AtuGST4 and AtuGST8 use hydrophobic ‘lock–and–key’-like motifs whereas the dimer interface of AtuGST2, AtuGST5, AtuGST6 and AtuGST7 is dominated by polar interactions. These results suggested that AtuGSTs could be involved in a broad range of biological functions including stress tolerance and detoxification of toxic compounds.

Proces prekaryzacji pracy - nowe kierunki zmian społeczno-gospodarczych w świecie

Wydział Nauk Politycznych i Dziennikarstwa

Administrative justice and the control of bureaucratic decision-making: A study investigating how decision-makers in local authority education departments respond to the work of redress mechanisms

This socio-legal thesis has explored the factors responsible for explaining whether and how redress mechanisms control bureaucratic decision-making. The research considered the three principal institutions of administrative justice: courts, tribunals, and ombudsman schemes. The field setting was the local authority education area and the thesis examined bureaucratic decision-making about admissions to school, home-to-school transport, and Special Educational Needs (SEN). The thesis adopted a qualitative approach, using interviews and documentary research, within a multiple embedded case study design. The intellectual foundations of the research were inter-disciplinary, cutting across law, socio-legal studies, public administration, organization studies, and social policy. The thesis drew on these scholarly fields to explore the nature of bureaucratic decision-making, the extent to which it can be controlled and the way that learning occurs in bureaucracies and, finally, the extent to which redress mechanisms might exercise control. The concept of control was studied across all its dimensions – in relation both to ex post control in specific cases and the more challenging notion of ex ante or structuring control. The aim of the thesis was not to measure the prevalence of bureaucratic control by redress mechanisms, but to understand the factors that might explain its presence or absence in a particular area. The findings of the research have allowed for a number of analytical refinements and extensions to be made to existing theoretical and empirical understandings. 14 factors, along with 87 supporting propositions, have been set out with the aim of making empirically derived suggestions which can be followed up in future research. In terms of the thesis’ contribution to existing knowledge, its comparative focus and its emphasis on the broad notion of control offered the potential for new insights to be developed. Overall, the thesis claims to have made three contributions to the conceptual framework for understanding the exercise of control by redress mechanisms: it emphasizes the importance of ‘feedback’ in relation to the nature of the cases referred to redress mechanisms; it calls attention to the structure of bureaucratic decision-making as well as its normative character; and it discusses how the operational modes of redress mechanisms relate to their control functions.

PI Toolset Methodology for Virtual Enterprise Performance Assessment and Governance

Part 16: Performance Measurement Systems

Learning English is Fun! Incidental vocabulary acquisition of music and traditional class ninth graders via listening to pop songs

Finnish youth are constantly exposed to music and lyrics in English in their free time. It is likely that this has a positive effect on vocabulary learning. Learning vocabulary while simultaneously accompanied with melodies is likely to result in better learning outcomes. The present thesis covers a study on the vocabulary learning of traditional and music class ninth graders in a south-western upper comprehensive school in Finland, mainly concentrating on vocabulary learning as a by-product of listening to pop music and learning vocabulary through semantic priming. The theoretical background presents viable linguistic arguments and theories, which provide clarity for why it would be possible to learn English vocabulary via listening to pop songs. There is conflicting evidence on the benefits of music on vocabulary learning, and this thesis sets out to shed light on the situation. Additionally, incorporating pop music in English classes could assist in decreasing the gap between real world English and school English. The thesis is a mixed method research study consisting of both quantitative and qualitative research materials. The methodology comprises vocabulary tests both before and after pop music samples and a background questionnaire filled by students. According to the results, all students reported liking listening to music and they clearly listened to English pop music the most. A statistically significant difference was found when analysing the results of the differences in pre- and post-vocabulary tests. However, the traditional class appeared to listen to mainstream pop music more than the students in the music class, and thus it seems likely that the traditional class benefited more from vocabulary learning occurring via listening to pop songs. In conclusion, it can be established that it is possible to learn English vocabulary via listening to pop songs and that students wish their English lectures would involve more music-related vocabulary exercises in the future. Thus, when it comes to school learning, pop songs should be utilised in vocabulary learning, which could also in turn result in more diverse learning and the students could, more easily than before, relate to the themes and topics of the lectures. Furthermore, with the help of pop songs it would be possible to decrease the gap between school English and real-world English.

Optimal resource allocation and budgeting in libraries

Libraries since their inception 4000 years ago have been in a process of constant change. Although, changes were in slow motion for centuries, in the last decades, academic libraries have been continuously striving to adapt their services to the ever-changing user needs of students and academic staff. In addition, e-content revolution, technological advances, and ever-shrinking budgets have obliged libraries to efficiently allocate their limited resources among collection and services. Unfortunately, this resource allocation is a complex process due to the diversity of data sources and formats required to be analyzed prior to decision-making, as well as the lack of efficient integration methods. The main purpose of this study is to develop an integrated model that supports libraries in making optimal budgeting and resource allocation decisions among their services and collection by means of a holistic analysis. To this end, a combination of several methodologies and structured approaches is conducted. Firstly, a holistic structure and the required toolset to holistically assess academic libraries are proposed to collect and organize the data from an economic point of view. A four-pronged theoretical framework is used in which the library system and collection are analyzed from the perspective of users and internal stakeholders. The first quadrant corresponds to the internal perspective of the library system that is to analyze the library performance, and costs incurred and resources consumed by library services. The second quadrant evaluates the external perspective of the library system; user’s perception about services quality is judged in this quadrant. The third quadrant analyses the external perspective of the library collection that is to evaluate the impact of the current library collection on its users. Eventually, the fourth quadrant evaluates the internal perspective of the library collection; the usage patterns followed to manipulate the library collection are analyzed. With a complete framework for data collection, these data coming from multiple sources and therefore with different formats, need to be integrated and stored in an adequate scheme for decision support. A data warehousing approach is secondly designed and implemented to integrate, process, and store the holistic-based collected data. Ultimately, strategic data stored in the data warehouse are analyzed and implemented for different purposes including the following: 1) Data visualization and reporting is proposed to allow library managers to publish library indicators in a simple and quick manner by using online reporting tools. 2) Sophisticated data analysis is recommended through the use of data mining tools; three data mining techniques are examined in this research study: regression, clustering and classification. These data mining techniques have been applied to the case study in the following manner: predicting the future investment in library development; finding clusters of users that share common interests and similar profiles, but belong to different faculties; and predicting library factors that affect student academic performance by analyzing possible correlations of library usage and academic performance. 3) Input for optimization models, early experiences of developing an optimal resource allocation model to distribute resources among the different processes of a library system are documented in this study. Specifically, the problem of allocating funds for digital collection among divisions of an academic library is addressed. An optimization model for the problem is defined with the objective of maximizing the usage of the digital collection over-all library divisions subject to a single collection budget. By proposing this holistic approach, the research study contributes to knowledge by providing an integrated solution to assist library managers to make economic decisions based on an “as realistic as possible” perspective of the library situation.

Physiological and morphological responses to feeding in broad-nosed caiman (Caiman latirostris)

Broad nosed caiman are ectotherm sauropsids that naturally experience long fasting intervals. We have studied the postprandial responses by measuring oxygen consumption using respirometry, the size changes of the duodenum, the distal small intestine, and the liver, using repeated non-invasive ultrasonography, and by investigating structural changes on the level of tissues and cells by using light- and electron microscopy. The caimans showed the same rapid and reversible changes of organ size and identical histological features, down to the ultrastructure level, as previously described for other ectothermic sauropsids. We found a configuration change of the mucosa epithelium from pseudostratified during fasting to single layered during digestion, in association with hypertrophy of enterocytes by loading them with lipid droplets. Similar patterns were also found for the hepatocytes of the liver. By placing the results of our study in comparative relationship and by utilizing the phylogenetic bracket of crocodiles, birds and squamates, we suggest that the observed features are plesiomorphic characters of sauropsids. By extending the comparison to anurans, we suggest that morphological and physiological adjustments to feeding and fasting described here may have been a character of early tetrapods. In conclusion, we suggest that the ability to tolerate long fasting intervals and then swallow a single large meal as described for many sit-an-wait foraging sauropsids is a functional feature that was already present in ancestral tetrapods.

Acute lethal and sublethal effects of neem leaf extract on the neotropical freshwater fish Prochilodus lineatus

The aim of this study was to determine the toxicity of the aqueous extract of neem leaves, a product extensively used in fish-farms as alternative for the control of fish parasites and fish fry predators, for the neotropical fish Prochilodus lineatus. The 24 It LC(50) of neem leaf extract for juveniles P lineatus was estimated as 4.8 g L(-1); the fish were then exposed for 24 h to 2.5, 5.0 and 7.5 g L(-1) or only clean water (control). Plasma glucose levels were higher in fish exposed to 2.5 g L(-1) and 5.0 g L(-1) neem extract, relative to control, indicating a typical stress response. Neem extract did not interfere with the osmoregulating capacity of the fish, as their plasma sodium, chloride, total protein and osmolarity did not change. The presence of the biopesticide interfered with the antioxidant defense system of P. lineatus, as there was a decrease in liver catalase activity at all neem concentrations and the detoxifying enzyme glutathione-S-transferase was activated in fish exposed to 5.0 g L(-1). Fish exposed to all neem extract concentrations exhibited damaged gill and kidney tissue. These results indicate that although neem extract is less toxic to P. lineatus than other synthetic insecticides used in fish-farming it does cause functional and morphological changes in this fish species. (c) 2006 Elsevier B.V. All rights reserved.

Respiratory metabolism of Camponotus rufipes ants: Brood and adults

The respiratory metabolism of immature forms (eggs, larvae, prepupae and pupae) of Camponotus rufipes (Hymenoptera: Formicidae) was studied at 25 degrees C, using a Warburg respirometer. Mean respiratory rates (mu l O gamma mg(-1) live weight.hr(-1)) for eggs, first instars, second instars, third instars, fourth instars, prepupae, and pupae were respectively: 2.53, 5.07, 1.23, 0.32, 0.22, 0.19 and 0.13. Adult workers with body mass between 20 and 30 mg had a mean respiratory rate of 0.43. The high respiratory rate in first instars probably reflects, besides the size influence, the metabolic costs of differentiation that occurs in this phase. (C) 1998 Published by Elsevier B.V.

Organisational cognitive neuroscience drives theoretical progress, or: the curious case of the straw man murder

In this critical essay, we respond to Lindebaum’s (2016) argument that neuroscientific methodologies and data have been accepted prematurely in proposing novel management theory. We acknowledge that building new management theories requires firm foundations. We also find his distinction between demand and supply side forces helpful as an analytical framework identifying the momentum for the contemporary production of management theory. Nevertheless, some of the arguments Lindebaum (2016) puts forward, on closer inspection, can be contested, especially those related to the supply side of organizational cognitive neuroscience (OCN) research: fMRI data, motherhood statements and ethical concerns. We put forward a more positive case for OCN methodologies and data, as well as clarifying exactly what OCN really means, and its consequences for the development of strong management theory.

Literatura y pensamiento en Francia. Un ejemplo : el panfleto

First of all, with this dissertation we have set as principal goal to define and to characterize the pamphlet in a French-speaking context. To do this, we will try to delimit this type of text and to demonstrate that in many occasions this term is used for classifying documents whose principal characteristic is the polemic. In the approach to this dissertation, we have mainly found two problems: on the one hand to define what a pamphlet is and to demarcate it from other similar texts both from the point of view of the content as well as the form; and on the other hand, to try to discern if the pamphlet can be included under the title of Literature and if it constitutes a literary genre for itself. To do so, we will structure our investigation in several chapters gathered in two clearly delimited parts. At the end of each of them, we will draw some brief conclusions. The first part will be focused on the study of theoretical notions. We will approach this reflection from different perspectives: etymological, comparative, literary and generic. To do this, we will approach the theories of those schools or of those authors who are more convenient to our purpose, so both our matter of study and the areas where we want to study it, raise many problems of interpretation and classification. Our aim is not to do a study on the Literature concepts or on the Literary Genre, so we will simplify those theories that are related to its main characteristics...

Animação sociocultural em lares de idosos do concelho da Guarda

Dissertação Trabalho de Projeto apresentado à Escola Superior de Educação do Instituto Politécnico de Castelo Branco para cumprimento dos requisitos necessários à obtenção do grau de Mestre em Gerontologia Social.

Purity of Blood and the Curial Market in Iberian Cathedrals

This article analyses the dynamics of accessing cathedral chapters with purity of blood statutes in the Iberian Peninsula, from a comparative perspective, and the interaction with the phenomenon of commodification of ecclesiastical benefices. This phenomenon created a Curial market that was open to any applicant with sufficient economic capacity, regardless of their ancestry. What was the result of the clash between the Iberian concept of purity and the commodified Roman reality?

Nanofibrous-structured polymers and their engineering applications

Monolithic materials cannot always satisfy the demands of today’s advanced requirements. Only by combining several materials at different length-scales, as nature does, the requested performances can be met. Polymer nanocomposites are intended to overcome the common drawbacks of pristine polymers, with a multidisciplinary collaboration of material science with chemistry, engineering, and nanotechnology. These materials are an active combination of polymers and nanomaterials, where at least one phase lies in the nanometer range. By mimicking nature’s materials is possible to develop new nanocomposites for structural applications demanding combinations of strength and toughness. In this perspective, nanofibers obtained by electrospinning have been increasingly adopted in the last decade to improve the fracture toughness of Fiber Reinforced Plastic (FRP) laminates. Although nanofibers have already found applications in various fields, their widespread introduction in the industrial context is still a long way to go. This thesis aims to develop methodologies and models able to predict the behaviour of nanofibrous-reinforced polymers, paving the way for their practical engineering applications. It consists of two main parts. The first one investigates the mechanisms that act at the nanoscale, systematically evaluating the mechanical properties of both the nanofibrous reinforcement phase (Chapter 1) and hosting polymeric matrix (Chapter 2). The second part deals with the implementation of different types of nanofibers for novel pioneering applications, trying to combine the well-known fracture toughness enhancement in composite laminates with improving other mechanical properties or including novel functionalities. Chapter 3 reports the development of novel adhesive carriers made of nylon 6,6 nanofibrous mats to increase the fracture toughness of epoxy-bonded joints. In Chapter 4, recently developed rubbery nanofibers are used to enhance the damping properties of unidirectional carbon fiber laminates. Lastly, in Chapter 5, a novel self-sensing composite laminate capable of detecting impacts on its surface using PVDF-TrFE piezoelectric nanofibers is presented.

Speed, legislatures and international treaties: Evidence from Mercosur

What causes faster or slower procedures in the parliaments when considering international treaties? This question motivates the current research, which aims to understand how the nature of coalitions influence the duration of the legislative processes. For this, the analysis covers all the treaties signed by Mercosur between 1991 and 2021 and the internalisation processes in four member states (Argentina, Brazil, Paraguay and Uruguay). It observes how long each parliament took to approve the treaties and which was the effect of political and economic variables. A mixed-methods approach was adopted for the empirical research, combining Survival Analysis, Qualitative Comparative Analysis and Process Tracing. While the quantitative work investigates all the cases, the qualitative study illuminates the enlargement of Mercosur, with in-depth analysis of the Paraguayan approval of the Venezuelan and Bolivian accessions. This study provides important insights into the role of national legislatures in the Latin American regionalism, concluding that the government-opposition cleavage drives the parliamentarians’ behaviour on the topic of regional integration. The study also contributes to the field Mercosur studies with the characterisation of the treaties ratified domestically, by undertaking a longitudinal analysis at the 30th anniversary of the bloc.