NMR structural analysis of cardiac troponin C: Monitoring conformational changes induced by binding calcium, troponin I, and calcium -sensitizing drugs

Contraction of cardiac muscle is regulated through the Ca2+ dependent protein-protein interactions of the troponin complex (Tn). The critical role cardiac troponin C (cTnC) plays as the Ca2+ receptor in this complex makes it an attractive target for positive inotropic compounds. In this study, the ten Met methyl groups in cTnC, [98% 13C ϵ]-Met cTnC, are used as structural markers to monitor conformational changes in cTnC and identify sites of interaction between cTnC and cardiac troponin I (cTnI) responsible for the Ca2+ dependent interactions. In addition the structural consequences that a number of Ca2+-sensitizing compounds have on free cTnC and the cTnC·cTnI complex were characterized. Using heteronuclear NMR experiments and monitoring chemical shift changes in the ten Met methyl 1H-13C correlations in 3Ca2+ cTnC when bound to cTnI revealed an anti-parallel arrangement for the two proteins such that the N-domain of cTnI interacts with the C-domain of cTnC. The large chemical shifts in Mets-81, -120, and -157 identified points of contact between the proteins that include the C-domain hydrophobic surface in cTnC and the A, B, and D helical interface located in the regulatory N-domain of cTnC. TnI association [cTnI(33–80), cTnI(86–211), or cTnI(33–211)] was found also to dramatically reduce flexibility in the D/E central linker of cTnC as monitored by line broadening in the Met 1H- 13C correlations of cTnC induced by a nitroxide spin label, MTSSL, covalently attached to cTnC at Cys 84. TnI association resulted in an extended cTnC that is unlike the compact structure observed for free cTnC. The Met 1H-13C correlations also allowed the binding characteristics of bepridil, TFP, levosimendan, and EMD 57033 to the apo, 2Ca2+, and Ca2+ saturated forms of cTnC to be determined. In addition, the location of drug binding on the 3Ca2+cTnC·cTnI complex was identified for bepridil and TFP. Use of a novel spin-labeled phenothiazine, and detection of isotope filtered NOEs, allowed identification of drug binding sites in the shallow hydrophobic cup in the C-terminal domain, and on two hydrophobic surfaces on N-regulatory domain in free 3Ca2+ cTnC. In contrast, only one N-domain drug binding site exists in 3Ca2+ cTnC·cTnI complex. The methyl groups of Met 45, 60 and 80, which are grouped in a hydrophobic patch near site II in cTnC, showed the greatest change upon titration with bepridil or TFP, suggesting that this is a critical site of drug binding in both free cTnC and when associated with cTnI. The strongest NOEs were seen for Met-60 and -80, which are located on helices C and D, respectively, of Ca2+ binding site II. These results support the conclusion that the small hydrophobic patch which includes Met-45, -60, and -80 constitutes a drug binding site, and that binding drugs to this site will lead to an increase in Ca2+ binding affinity of site II while preserving maximal cTnC activity. Thus, the subregion in cTnC makes a likely target against which to design new and selective Ca2+-sensitizing compounds. ^

End -of -life planning and decision -making: Implications for adults with mental retardation

This research focused on the topic of end-of-life planning and decision-making for adults affected by mental retardation. Adults with mental retardation have unique challenges in this regard, including difficulty communicating their wishes without assistance and diminished decision-making skills. The primary research objective was to identify factors that can affect opportunities for adults with mental retardation in community-based services settings (and their advocates) to be involved in planning and deciding about their own end-of-life experience. ^ A descriptive qualitative inquiry was designed to explore issues related to death and dying, and the notion of end-of-life planning, from the perspective of adults with mental retardation who receive publicly-funded community services ("clients") and family members of individuals who receive such services. Study participants were recruited from a single mental retardation service provider in a large urban setting (the "Agency"). Sixteen clients and 14 families of Agency clients took part. Client data collection was accomplished through face-to-face interviews, focus group meetings, and record reviews; family members were involved in a face-to-face interview only. ^ An initial coding scheme was developed based upon literature and policy reviews, and themes related to the research questions. Analysis involved extracting data from transcripts and records and placing it into appropriate thematic categories, building support for each theme with the accumulated data. Coding themes were modified to accommodate new data when it challenged existing themes. ^ Findings suggest that adults with mental retardation do have the requisite knowledge, interest, and ability to participate in decisions about their end-of-life experience and handling of affairs. Siblings are overwhelmingly the chosen future surrogates and they (or their children) will likely be the end-of-life advocates for their brothers and sisters affected by mental retardation. Findings further point to a need for increased awareness, accurate information, and improved communication about end-of-life issues, both in general and particular to adults affected by mental retardation. Also suggested by the findings is a need to focus on creating accommodations and adaptations that can best uncover a person's authentic views on life and death and related end-of-life preferences. Practical implications and suggestions for further research are also discussed. ^

The attention/support to students in the socio-labor transition

In the present paper I intend to put forward some inquiries regarding the socio-labor guidance and transition of the young people in the co-ordinates of contemporary society. The outstanding paradox in which some young people are immersed when trying to get a first, second, third employment is also analyzed: "The job of looking for a job". This is a common experience shared by many young people when they try to find a place in the complex labor panorama. Looking for a job is a recurrent situation in contemporary society, given the high dose of precarious employment in which the productive world moves. Life-long jobs belong to the past and what is normal is that people strive to enter the competitive labor market repeatedly. Moreover, the paper offers the partial results of a larger research project that approaches tutorship, decision taking and expectations before the academic-labor future of the studentship in the last years of secondary school. Finally, I suggest some insights in the line that it is not longer feasible to work with old-fashioned guidance, trying to channel people's vocation and offering them information so that they get a job that provides them with the happiness of "doing what you like". Behind these ideas lay the normal biographies, the predictable itineraries. Labor market complexity makes the transition to working life very difficult. Once immersed in it, biographies and labor itineraries are constructed and reconstructed at the rhythm of the changing fortunes of times.

Conflictividad y resistencias en los márgenes de La Pampa (1990-2012)

Las formas de producción de territorios materiales e inmateriales entran en tensión y se yuxtaponen cuando espacios dominados por la territorialidad campesina son revalorizados por el capital y buscan imponer su espacialidad. En el extremo oeste pampeano, en el período 1990-2012, además de las nuevas racionalidades y prácticas de los campesinos, avanzaron el proceso de mercantilización y el accionar de productores-empresarios extralocales. En este contexto, emergieron los conflictos por el uso y la apropiación de los recursos, el modo de vida y la construcción social del espacio. La organización de las familias que ejercen actos posesorios para conservar la forma de manejo de los "campos abiertos" y el control del territorio dio origen a la conformación de estrategias de lucha colectivas. Ante los conflictos generados por el uso del monte y el achicamiento de los campos de los últimos diez años por la llegada de nuevos agentes que cercan las propiedades y desarticulan los circuitos de pastoreo, muchas familias del paraje comenzaron a organizarse con el apoyo de organizaciones urbanas para denunciar la reducción del espacio de pastoreo compartido. Con la finalidad de analizar los cambios y reconstruir las estrategias de organización campesinas, utilizamos distintos materiales empíricos en los que se combinan el análisis de estadísticas con entrevistas en profundidad e interpretación de diversos documentos

Conflictividad y resistencias en los márgenes de La Pampa (1990-2012)

Las formas de producción de territorios materiales e inmateriales entran en tensión y se yuxtaponen cuando espacios dominados por la territorialidad campesina son revalorizados por el capital y buscan imponer su espacialidad. En el extremo oeste pampeano, en el período 1990-2012, además de las nuevas racionalidades y prácticas de los campesinos, avanzaron el proceso de mercantilización y el accionar de productores-empresarios extralocales. En este contexto, emergieron los conflictos por el uso y la apropiación de los recursos, el modo de vida y la construcción social del espacio. La organización de las familias que ejercen actos posesorios para conservar la forma de manejo de los "campos abiertos" y el control del territorio dio origen a la conformación de estrategias de lucha colectivas. Ante los conflictos generados por el uso del monte y el achicamiento de los campos de los últimos diez años por la llegada de nuevos agentes que cercan las propiedades y desarticulan los circuitos de pastoreo, muchas familias del paraje comenzaron a organizarse con el apoyo de organizaciones urbanas para denunciar la reducción del espacio de pastoreo compartido. Con la finalidad de analizar los cambios y reconstruir las estrategias de organización campesinas, utilizamos distintos materiales empíricos en los que se combinan el análisis de estadísticas con entrevistas en profundidad e interpretación de diversos documentos

The attention/support to students in the socio-labor transition

In the present paper I intend to put forward some inquiries regarding the socio-labor guidance and transition of the young people in the co-ordinates of contemporary society. The outstanding paradox in which some young people are immersed when trying to get a first, second, third employment is also analyzed: "The job of looking for a job". This is a common experience shared by many young people when they try to find a place in the complex labor panorama. Looking for a job is a recurrent situation in contemporary society, given the high dose of precarious employment in which the productive world moves. Life-long jobs belong to the past and what is normal is that people strive to enter the competitive labor market repeatedly. Moreover, the paper offers the partial results of a larger research project that approaches tutorship, decision taking and expectations before the academic-labor future of the studentship in the last years of secondary school. Finally, I suggest some insights in the line that it is not longer feasible to work with old-fashioned guidance, trying to channel people's vocation and offering them information so that they get a job that provides them with the happiness of "doing what you like". Behind these ideas lay the normal biographies, the predictable itineraries. Labor market complexity makes the transition to working life very difficult. Once immersed in it, biographies and labor itineraries are constructed and reconstructed at the rhythm of the changing fortunes of times.

Conflictividad y resistencias en los márgenes de La Pampa (1990-2012)

Las formas de producción de territorios materiales e inmateriales entran en tensión y se yuxtaponen cuando espacios dominados por la territorialidad campesina son revalorizados por el capital y buscan imponer su espacialidad. En el extremo oeste pampeano, en el período 1990-2012, además de las nuevas racionalidades y prácticas de los campesinos, avanzaron el proceso de mercantilización y el accionar de productores-empresarios extralocales. En este contexto, emergieron los conflictos por el uso y la apropiación de los recursos, el modo de vida y la construcción social del espacio. La organización de las familias que ejercen actos posesorios para conservar la forma de manejo de los "campos abiertos" y el control del territorio dio origen a la conformación de estrategias de lucha colectivas. Ante los conflictos generados por el uso del monte y el achicamiento de los campos de los últimos diez años por la llegada de nuevos agentes que cercan las propiedades y desarticulan los circuitos de pastoreo, muchas familias del paraje comenzaron a organizarse con el apoyo de organizaciones urbanas para denunciar la reducción del espacio de pastoreo compartido. Con la finalidad de analizar los cambios y reconstruir las estrategias de organización campesinas, utilizamos distintos materiales empíricos en los que se combinan el análisis de estadísticas con entrevistas en profundidad e interpretación de diversos documentos

The attention/support to students in the socio-labor transition

In the present paper I intend to put forward some inquiries regarding the socio-labor guidance and transition of the young people in the co-ordinates of contemporary society. The outstanding paradox in which some young people are immersed when trying to get a first, second, third employment is also analyzed: "The job of looking for a job". This is a common experience shared by many young people when they try to find a place in the complex labor panorama. Looking for a job is a recurrent situation in contemporary society, given the high dose of precarious employment in which the productive world moves. Life-long jobs belong to the past and what is normal is that people strive to enter the competitive labor market repeatedly. Moreover, the paper offers the partial results of a larger research project that approaches tutorship, decision taking and expectations before the academic-labor future of the studentship in the last years of secondary school. Finally, I suggest some insights in the line that it is not longer feasible to work with old-fashioned guidance, trying to channel people's vocation and offering them information so that they get a job that provides them with the happiness of "doing what you like". Behind these ideas lay the normal biographies, the predictable itineraries. Labor market complexity makes the transition to working life very difficult. Once immersed in it, biographies and labor itineraries are constructed and reconstructed at the rhythm of the changing fortunes of times.

Conflictividad y resistencias en los márgenes de La Pampa (1990-2012)

Las formas de producción de territorios materiales e inmateriales entran en tensión y se yuxtaponen cuando espacios dominados por la territorialidad campesina son revalorizados por el capital y buscan imponer su espacialidad. En el extremo oeste pampeano, en el período 1990-2012, además de las nuevas racionalidades y prácticas de los campesinos, avanzaron el proceso de mercantilización y el accionar de productores-empresarios extralocales. En este contexto, emergieron los conflictos por el uso y la apropiación de los recursos, el modo de vida y la construcción social del espacio. La organización de las familias que ejercen actos posesorios para conservar la forma de manejo de los "campos abiertos" y el control del territorio dio origen a la conformación de estrategias de lucha colectivas. Ante los conflictos generados por el uso del monte y el achicamiento de los campos de los últimos diez años por la llegada de nuevos agentes que cercan las propiedades y desarticulan los circuitos de pastoreo, muchas familias del paraje comenzaron a organizarse con el apoyo de organizaciones urbanas para denunciar la reducción del espacio de pastoreo compartido. Con la finalidad de analizar los cambios y reconstruir las estrategias de organización campesinas, utilizamos distintos materiales empíricos en los que se combinan el análisis de estadísticas con entrevistas en profundidad e interpretación de diversos documentos

Case Study of Applied LIP Approach/Activities in the Philippines The Training Services Enhancement Project for Rural Life Improvement (TSEP-RLI) Experience

TSEP-RLI was a technical cooperation project jointly conducted by GOP thru DA-Agricultural Training Institute (ATI) and GOJ thru JICA aimed at institutionalizing the training program for Rural Life Improvement (RLI) at the (ATI). As expected, farmers, fisherfolk, women, youth and extension agents were provided with efficient and effective training services from ATI leading to the improvement of quality of life in the rural areas through efforts of human resource development. The ATI- Bohol was chosen as the model center where participatory trials and various activities of the project were undertaken for five years. These activities were participatory surveys and data collection of on-farm and off-farm productive activities; planning workshop for RLI; feedbacking of survey results and action plans to the community and the Local Government Units (LGUs), and signing of Memorandum of Agreement between the Project and participating LGUs. The above activities were done to facilitate the planning and development of most effective and necessary rural life improvement activities, to confirm the willingness of the people to support and participate and to formalize the partnership between the Project and the LGUs. Since the concept of rural life covers a vast range of activities, a consensus had been reached that the total aspects of rural life be grasped in three spheres, namely, Production & Livelihood (P/L), Rural Living Condition (RLC) and Community Environment (C/E). The RLI for Ubi (Yam) Growers was one of the pilot activities undertaken in two pilot barangays and the target beneficiaries were members of the Rural Improvement Club (RIC- a group of organized women) with the LGU of the Municipality of Corella as the implementing partner. During the planning workshop, the barangay residents articulated their desire to promote production and processing of ubi (sphere on P/L - as the entry point), lack of nutritious food was one of the identified problem (sphere on RLC- expansion point) and environmental degradation such as deforestation, and soil erosion was another problem articulated by the community people (sphere on C/E- expansion point). Major activities that were undertaken namely, Ubi cooking contest, cooking/processing seminar, training courses on entrepreneurial development, ubi production and storage technology, packaging and product design, human resource development and simplified bookkeeping motivated the beneficiaries as well as developed and enhanced their skills & capabilities while strengthening their associations. Their participation to the 5 ubi festivals and other related activities had brought some impacts on their economic and rural life improvement activities. The seven principles of TSEP-RLI include the participatory process, holistic approach, dialogical approach, bottom -up training needs assessment, demand-driven approach, cost sharing approach and collaborative implementation with other agencies including LGUs and the community.

Public support for the deployment of next generation access networks-Common themes, methodological caveats and further research

Next generation telecommunications infrastructures are considered as a principal example of a new technology for sustainable economic growth. From their deployment it is expected that a wealth of innovations – hopefully converted into economic growth – new sources of employment and improved quality of life will result. In line with these prospects, public administrations at supranational, national, regional and local levels have encouraged the development of these new infrastructures. Moreover, in times of economic crisis, public assistance to deploy such networks encompasses the promise of placing a weak economy on the road to prosperity. However, such arguments and political claims clearly require rigorous assessment. In particular, any such assessment must adequately address the appropriate form of modelling that best captures key elements for identifiable progress from next generation access networks (NGAN).

PubDNA Finder in a Nutshell. Searching the Life Sciences Literature with Sequences of Nucleic Acids

Biomedical researchers and clinicians working with molecular technologies in routine clinical practice often need to review the available literature to gather information regarding specific sequences of nucleic acids. This includes, for instance, finding articles related to a concrete DNA sequence, or identifying empirically-validated primer/probe sequences to evaluate the presence of different micro-organisms. Unfortunately, these hard and time-consuming tasks often need to be manually performed by researchers themselves since no publicly available biomedical literature search engine, e.g. PubMed, PubMed Central (PMC), etc., provides the required search functionalities. In this article, we describe PubDNA Finder, a web service that enables users to perform advanced searches on PubMed Central-indexed full text articles with sequences of nucleic acids

A Multi-agent System for Emergency Decision Support

This paper describes the multi-agent organization of a computer system that was designed to assist operators in decision making in the presence of emergencies. The application was developed for the case of emergencies caused by river floods. It operates on real-time receiving data recorded by sensors (rainfall, water levels, flows, etc.) and applies multi-agent techniques to interpret the data, predict the future behavior and recommend control actions. The system includes an advanced knowledge based architecture with multiple symbolic representation with uncertainty models (bayesian networks). This system has been applied and validated at two particular sites in Spain (the Jucar basin and the South basin).

Engineering Agent Systems for Decision Support

This paper discusses how agent technology can be applied to the design of advanced Information Systems for Decision Support. In particular, it describes the different steps and models that are necessary to engineer Decision Support Systems based on a multiagent architecture. The approach is illustrated by a case study in the traffic management domain.

A Process Asset Library to support Software Process Improvement in Small Settings

A main factor to the success of any organization process improvement effort is the Process Asset Library implementation that provides a central database accessible by anyone at the organization. This repository includes any process support materials to help process deployment. Those materials are composed of organization's standard software process, software process related documentation, descriptions of the software life cycles, guidelines, examples, templates, and any artefacts that the organization considers useful to help the process improvement. This paper describe the structure and contents of the Web-based Process Asset Library for Small businesses and small groups within large organizations. This library is structured using CMMI as reference model in order to implement those Process Areas described by this model.