Fate of linear and supercoiled multinucleosomic templates during transcription.

Electron microscopy was used to monitor the fate of reconstituted nucleosome cores during in vitro transcription of long linear and supercoiled multinucleosomic templates by the prokaryotic T7 RNA polymerase and the eukaryotic RNA polymerase II. Transcription by T7 RNA polymerase disrupted the nucleosomal configuration in the transcribed region, while nucleosomes were preserved upstream of the transcription initiation site and in front of the polymerase. Nucleosome disruption was independent of the topology of the template, linear or supercoiled, and of the presence or absence of nucleosome positioning sequences in the transcribed region. In contrast, the nucleosomal configuration was preserved during transcription from the vitellogenin B1 promoter with RNA polymerase II in a rat liver total nuclear extract. However, the persistence of nucleosomes on the template was not RNA polymerase II-specific, but was dependent on another activity present in the nuclear extract. This was demonstrated by addition of the extract to the T7 RNA polymerase transcription reaction, which resulted in retention of the nucleosomal configuration. This nuclear activity, also found in HeLa cell nuclei, is heat sensitive and could not be substituted by nucleoplasmin, chromatin assembly factor (CAF-I) or a combination thereof. Altogether, these results identify a novel nuclear activity, called herein transcription-dependent chromatin stabilizing activity I or TCSA-I, which may be involved in a nucleosome transfer mechanism during transcription.

Transapical aortic 'valve-in-valve' procedure for degenerated stented bioprosthesis.

Standard surgical aortic valve replacement with a biological prosthesis remains the treatment of choice for low- and mid-risk elderly patients (traditionally >65 years of age) suffering from severe symptomatic aortic valve stenosis or insufficiency, and for young patients with formal contraindications to long-lasting anticoagulation. Unfortunately, despite the fact that several technical improvements have noticeably improved the resistance of pericardial and bovine bioprostheses to leaflet calcifications and ruptures, the risk of early valve failure with rapid degeneration still exists, especially for patients under haemodialysis and for patients <60 years of age at the time of surgery. Until now, redo open heart surgery under cardiopulmonary bypass and on cardioplegic arrest was the only available therapeutic option in case of bioprosthesis degeneration, but it carried a higher surgical risk when elderly patients with severe concomitant comorbidities were concerned. Since a few years, the advent of new transcatheter aortic valve procedures has opened new horizons in cardiac surgery and, in particular, the possibility of implanting stented valves within the degenerated stented bioprosthesis, the so-called 'valve-in-valve' (VinV) concept, has become a clinical practice in experienced cardiac centres. The VinV procedure represents a minimally invasive approach dedicated to high-risk redo patients, and published preliminary reports have shown a success rate of 100% with absence of significant valvular leaks, acceptable transvalvular gradients and low complication rate. However, this procedure is not riskless and the most important concerns are about the size mismatch and the right positioning within the degenerated bioprosthesis. In this article, we review the limited available literature about VinV procedures, underline important technical details for the positioning and provide guidelines to prevent valve-prosthesis mismatch comparing the three sizes of the only commercially available transapical device, the Edwards Sapien, with the inner diameter of three of the most commonly used stented bioprostheses.

The P-median problem in a changing network: The case of Barcelona

In this paper a p--median--like model is formulated to address theissue of locating new facilities when there is uncertainty. Severalpossible future scenarios with respect to demand and/or the travel times/distanceparameters are presented. The planner will want a strategy of positioning thatwill do as ``well as possible'' over the future scenarios. This paper presents a discrete location model formulation to address this P--Medianproblem under uncertainty. The model is applied to the location of firestations in Barcelona.

Actividades e Tarefas Desenvolvidas na Embaixada de Cabo Verde em Portugal

O presente relatório inclui os resultados do estudo dos Sectores de Actividades da Embaixada de Cabo Verde em Portugal. Este trabalho decorreu em três etapas essências. A primeira foi leitura de um Manual de Procedimentos sobre a “Concepção e Implementação de um Sistema Administrativo e de Controlo Orçamental. A segunda centrou-se no Sector Administrativo, onde foram aplicados conhecimentos estatísticos nas diversas actividades realizadas. A última centrou-se no sector da Tesouraria e Contabilidade, onde adquiri conhecimentos básicos sobre o sector financeiro. O relatório encontra–se estruturado em 5 capítulos: O capítulo I fornece, de um modo sintético, uma panorâmica histórica da Embaixada de Cabo Verde em Portugal. O capítulo II, tem um carácter descritivo, ou seja descreve sumáriamente, os diversos sectores de actividades. As mais variadas actividades que giram em torno do Serviço Administrativo e Financeiro, justificaram um tratamento detalhado no capítulo III. A questão da comunidade cabo-verdiana, residente em Portugal, os softwares de Contabilidade Primavera, e Gestão de Requisição são abordadas no capítulo IV, sendo analisado o posicionamento da população de origem cabo-verdiana que reside em Portugal, aplicando certos casos e exemplos nos próprios softwares. O capítulo V sintetiza e conjuga as principais conclusões parcelares que foram apresentadas nos diversos capítulos, incluindo também limitações e trabalho futuro. Por fim está organizado um conjunto de anexos compostos por: documentos da Embaixada, glossário financeiro, e scrip do software “R”, que serviram de suporte ao estudo.

Inovação e Tecnologia na Formação Agrícola – Agricultura de Precisão

O conceito de Agricultura de Precisão está normalmente associado à utilização de equipamento de alta tecnologia (seja hardware, no sentido genérico do termo, ou software) para avaliar, ou monitorizar, as condições numa determinada parcela de terreno, aplicando depois os diversos factores de produção (sementes, fertilizantes, fitofármacos, reguladores de crescimento, água, etc.) em conformidade. Tanto a monitorização como a aplicação diferenciada, ou à medida, exigem a utilização de tecnologias recentes, como os sistemas de posicionamento a partir de satélites (v.g. GPS - Global Positioning System), os Sistemas de Informação Geográfica (SIG) ou os sensores electrónicos, associados quer a reguladores automáticos de débito nas máquinas de distribuição quer a medidores de fluxo nas máquinas de colheita. A Agricultura de Precisão aparece, geralmente, associada a dois objectivos genéricos: o aumento do rendimento dos agricultores; e, a redução do impacte ambiental resultante da actividade agrícola. O primeiro destes objectivos pode, por sua vez, ser alcançado por duas vias distintas mas complementares: a redução dos custos de produção; e, o aumento da produtividade (e, por vezes, também da qualidade) das culturas. O cumprimento do segundo daqueles objectivos está relacionado com o rigor do controlo da aplicação dos factores de produção (sobretudo, produtos químicos, atendendo às externalidades ambientais negativas que lhes estão normalmente associadas), que deverá ser feita, tanto quanto possível, na justa medida das necessidades das plantas. De facto, se soubermos, por exemplo, que as necessidades de azoto em duas áreas distintas de uma mesma parcela de terreno não são iguais, por hipótese, em função dos resultados da análise de terras para as duas situações, teremos, do ponto de vista estritamente técnico e teórico, vantagem em fazer variar a quantidade de adubo azotado em conformidade. Deste modo, seríamos naturalmente tentados a aplicar mais adubo na área em que as necessidades ou o potencial produtivo são maiores, e a reduzir a quantidade na área em que o potencial produtivo ou as necessidades são menores, em vez de, como usualmente sucede, aplicar um valor médio e igual em toda a parcela. Com esta forma de actuar, não só aumentaríamos a produção, aplicando mais adubo onde este é necessário, como também reduziríamos os custos e o impacte ambiental da actividade, não aplicando adubo em excesso e precavendo a provável lixiviação do azoto. Na prática, tudo isto seria inquestionável, não fôra o caso de as tecnologias associadas à Agricultura de Precisão serem, quase sempre, complexas e caras. É exactamente por esta razão que o conceito não se encontra hoje em dia mais divulgado, nomeadamente no nosso país. Em primeiro lugar, só fará sentido recorrer à Agricultura de Precisão se os benefícios económicos daí decorrentes forem superiores ao investimento necessário à sua adopção; ora, infelizmente, são muito poucas as explorações, sobretudo em Portugal, com dimensão suficiente para, só por si, justificar ou viabilizar tais investimentos. Note-se que os investimentos a que nos referimos não passam apenas pela aquisição de determinados equipamentos (GPS, sensores, etc.), mas também pelo levantamento da situação de base e pela construção de um sistema de informação geograficamente referenciada. Em segundo lugar, existe ainda um longo caminho a percorrer pelas tecnologias de informação associadas, especialmente no que se refere à sua facilidade de uso e de integração no negócio, isto é, ainda é necessário um esforço razoável para efectuar a recolha e processamento da informação necessária à prática da agricultura de precisão, esforço este com custos muitas vezes inaceitáveis para o agricultor, nomeadamente quanto ao dispêndio do seu tempo/atenção. Em terceiro lugar, na hipótese de que os investimentos sejam rentáveis, é necessário que existam pessoas (agricultores e/ou técnicos) com conhecimentos suficientes para ajustar, desenvolver e utilizar estas tecnologias. O futuro, apesar de tudo, apresenta-se mais favorável. Por um lado, os equipamentos de alta tecnologia tendem a diminuir de preço, por vezes de forma muito marcada (um GPS de gama baixa, por exemplo, custava mais de 5000 euros há dez anos, existindo hoje à venda modelos similares por menos de 200 euros). Por outro, o nível educacional dos agricultores tem vindo a aumentar, existindo hoje cada vez mais estruturas de apoio técnico na agricultura. O nascimento e desenvolvimento de empresas especializadas no aluguer de máquinas e equipamentos agrícolas, que se tem vindo a registar nos últimos anos, pode igualmente contribuir para ultrapassar uma das maiores limitações à adopção destas tecnologias: a reduzida dimensão das explorações e os elevados custos de amortização daí decorrentes.

Transcatheter aortic valve implantation (TAVI): state of the art techniques and future perspectives.

Transcatheter aortic valve therapies are the newest established techniques for the treatment of high risk patients affected by severe symptomatic aortic valve stenosis. The transapical approach requires a left anterolateral mini-thoracotomy, whereas the transfemoral method requires an adequate peripheral vascular access and can be performed fully percutaneously. Alternatively, the trans-subclavian access has been recently proposed as a third promising approach. Depending on the technique, the fine stent-valve positioning can be performed with or without contrast injections. The transapical echo-guided stent-valve implantation without angiography (the Lausanne technique) relies entirely on transoesophageal echocardiogramme imaging for the fine stent-valve positioning and it has been proved that this technique prevents the onset of postoperative contrast-related acute kidney failure. Recent published reports have shown good hospital outcomes and short-term results after transcatheter aortic valve implantation, but there are no proven advantages in using the transfemoral or the transapical technique. In particular, the transapical series have a higher mean logistic Euroscore of 27-35%, a procedural success rate above 95% and a mean 30-day mortality between 7.5 and 17.5%, whereas the transfemoral results show a lower logistic Euroscore of 23-25.5%, a procedural success rate above 90% and a 30-day mortality of 7-10.8%. Nevertheless, further clinical trials and long-term results are mandatory to confirm this positive trend. Future perspectives in transcatheter aortic valve therapies would be the development of intravascular devices for the ablation of the diseased valve leaflets and the launch of new stent-valves with improved haemodynamic, different sizes and smaller delivery systems.

GPS analysis of human locomotion: further evidence for long-range correlations in stride-to-stride fluctuations of gait parameters.

During free walking, gait is automatically adjusted to provide optimal mechanical output and minimal energy expenditure; gait parameters, such as cadence, fluctuate from one stride to the next around average values. It was described that this fluctuation exhibited long-range correlations and fractal-like patterns. In addition, it was suggested that these long-range correlations disappeared if the participant followed the beep of metronome to regulate his or her pace. Until now, these fractal fluctuations were only observed for stride interval, because no technique existed to adequately analyze an extended time of free walking. The aim of the present study was to measure walking speed (WS), step frequency (SF) and step length (SL) with high accuracy (<1 cm) satellite positioning method (global positioning system or GPS) in order to detect long-range correlations in the stride-to-stride fluctuations. Eight participants walked 30 min under free and constrained (metronome) conditions. Under free walking conditions, DFA (detrended fluctuation analysis) and surrogate data tests showed that the fluctuation of WS, SL and SF exhibited a fractal pattern (i.e., scaling exponent alpha: 0.5 < alpha < 1) in a large majority of participants (7/8). Under constrained conditions (metronome), SF fluctuations became significantly anti-correlated (alpha < 0.5) in all participants. However, the scaling exponent of SL and WS was not modified. We conclude that, when the walking pace is controlled by an auditory signal, the feedback loop between the planned movement (at supraspinal level) and the sensory inputs induces a continual shifting of SF around the mean (persistent anti-correlation), but with no effect on the fluctuation dynamics of the other parameters (SL, WS).

A universal microscope manipulator

A modified and improved model of a mechanical manipulator for observation of pinned and mounted insects is described. This device allows movement of the observed object around three perpendicular axes in the field of vision at all magnifications of stereomicroscopes. The main improvement of this new model is positioning of the guiding knobs for rotating around two of the axes next to each other, allowing faster and easier manipulation of the studied object. Thus, one of the main advantages of this device is the possibility to rotate the specimen without the need to refocus. The device enables easily reaching a precession deviation in the intersection point of axes up to 0.5 mm in the process of assembling.

How to Preserve Renal Function in Transcatheter Aortic Valve Therapies

Background: Transcatheter aortic valve implantations (TAVI) are indicated in high risk patients requiring aortic valve replacement (AVR). However, CT-scans, coronary angiograms and intraoperative aortographies can induce contrast-related nephro-toxicity with a concrete risk of acute postoperative renal failure, especially in severely diseased patients. To prevent this complication, we routinely perform transapical (TA) TAVI guided by transesophageal echocardiogram and fluoroscopy without angiography. Material and Methods: From November 2008 to December 2009, 31 high-risk patients suffering from severe symptomatic aortic stenosis underwent TA-TAVI in our institution. The preoperative imaging assessment (cardiac CT-scan and coronary angiogram) was performed no less than 10 days before the TA-TAVI in all patients (to recover the renal function) with a low-dose protocol for injected contrast medium (equivalent to the patient's weight for the CT-scan). During the TA-TAVI, the stent-valve positioning was performed without any contrast injection. Results: 32 consecutive stent-valve were successfully positioned in 31 patients (mean age 80.76 8 8.3 years; mean EuroSCORE: 32.2 8 12.9%) through a transapical access (1 patient required 2 valves for valve embolisation). The mean preoperative creatinine and urea blood levels were 102.6 8 67.7 _ g/dl (range 53-339 _ g/dl) and 8.45 8 4.9 mmol/l, respectively. A chronic renal insufficiency affected 12 patients (38.7%) with 1 patient in pre-dialysis. Postoperatively, no patient developed acute myocardial infarction, atrio-ventricular block or acute renal insufficiency (mean creatinine level: 89.7 8 64.55 _ g/dl; urea level: 7.11 8 3.47 mmol/l) and the 30-days mortality was 9.67% (3 patients). Conclusion: Specific preoperative and intraoperative protocols that require lowdoses or absence of contrast medium are useful to preserve the renal function in high risk patients operated for TAVI.

Baerveldt aqueous shunt implantation into the ciliary sulcus

Purpose: Aqueous shunt implantation into the anterior chamber is associated with corneal decompensation in up to a third of eyes. Intracameral tube position may affect corneal endothelial cell loss. The authors set out to examine the efficacy and safety of Baerveldt shunt implantation into the ciliary sulcus combined with surgical peripheral iridectomy (SPI). Methods: One hundred eyes prospectively underwent Baerveldt shunt implantation into the cilliary sulcus combined with SPI, leaving a short intracameral tube length (1-2mm). Pre and post operative measures recorded included patient demographics, visual acuity, IOP, number of glaucoma medications (GMs) and all complications. Pre-existing corneal decompensation was recorded. Success was defined as IOP≤21mmHg and 20% reduction in IOP from baseline with or without GMs. Results: Mean age was 65.4 years (±20.4years). Mean follow-up was 10.8 months. Preoperatively IOP was 25.7mmHg (± 9.9mmHg), GMs were 2.9 (±1.2) and VA was 0.4 (±0.3). At one year postoperatively there was a significant drop in IOP (mean= 13.3mmHg (± 5.0mmHg); p<0.001) and number of GMs (mean= 1.3 (±1.4); p<0.001); and no significant change in VA (mean= 0.4 (±0.3); p=0.93). The success rate at one year was 83%. Complications were minor and non sight threatening (10%), there were no cases of postoperative corneal decompensation, tube blockage or iris/corneal-tube contact. Conclusions: The results demonstrate that placement of Baerveldt shunts into the ciliary sulcus with SPI is a safe and efficacious method of IOP reduction in comparison with standard shunt positioning in the anterior chamber. The intracameral tube position combined with SPI avoided tube-iris contact and corneal decompensation. Sulcus placement of aqueous shunts should be considered in pseudophakic eyes.

Development of a system for 3D high-resolution seismic reflection profiling on lakes

A high-resolution three-dimensional (3D) seismic reflection system for small-scale targets in lacustrine settings has been developed. Its main characteristics include navigation and shot-triggering software that fires the seismic source at regular distance intervals (max. error of 0.25 m) with real-time control on navigation using differential GPS (Global Positioning System). Receiver positions are accurately calculated (error < 0.20 m) with the aid of GPS antennas attached to the end of each of three 24-channel streamers. Two telescopic booms hold the streamers at a distance of 7.5 m from each other. With a receiver spacing of 2.5 m, the bin dimension is 1.25 m in inline and 3.75 m in crossline direction. To test the system, we conducted a 3D survey of about 1 km(2) in Lake Geneva, Switzerland, over a complex fault zone. A 5-m shot spacing resulted in a nominal fold of 6. A double-chamber bubble-cancelling 15/15 in(3) air gun (40-650 Hz) operated at 80 bars and 1 m depth gave a signal penetration of 300 m below water bottom and a best vertical resolution of 1.1 m. Processing followed a conventional scheme, but had to be adapted to the high sampling rates, and our unconventional navigation data needed conversion to industry standards. The high-quality data enabled us to construct maps of seismic horizons and fault surfaces in three dimensions. The system proves to be well adapted to investigate complex structures by providing non-aliased images of reflectors with dips up to 30 degrees.

Laparoscopic inguinal hernia repair with extraperitoneal double-mesh technique.

Totally extraperitoneal laparoscopic hernia repair is an efficient but technically demanding procedure. As mechanisms of hernia recurrence may be related to these technical difficulties, we have modified a previously described double-mesh technique in an effort to simplify the procedure. Extraperitoneal laparoscopic hernia repairs were performed in 82 male and 17 female patients having inguinal, femoral, and recurrent bilateral hernias. A standard propylene mesh measuring 15 x 15 cm was cut into two pieces of 4 x 15 cm and 11 x 15 cm. The smaller mesh was placed over both inguinal rings without splitting. The larger mesh was then inserted over the first mesh and stapled to low-risk zones, reinforcing the large-vessel area and the nerve transition zone. The mean procedure duration was 60 minutes for unilateral and 100 minutes for bilateral hernia repair. Patients were discharged from the hospital within 48 hours. The mean postoperative follow-up was 22 months, with no recurrences, neuralgia, or bleeding complications. Over a 2-year period, this technique was found to be satisfactory without recurrences or significant complications. In our hands, this technique was easier to perform: it allows for a less than perfect positioning of the meshes and avoids most of the stapling to crucial zones.

Extremity exposure in nuclear medicine: preliminary results of a European study.

The Work Package 4 of the ORAMED project, a collaborative project (2008-11) supported by the European Commission within its seventh Framework Programme, is concerned with the optimisation of the extremity dosimetry of medical staff in nuclear medicine. To evaluate the extremity doses and dose distributions across the hands of medical staff working in nuclear medicine departments, an extensive measurement programme has been started in 32 nuclear medicine departments in Europe. This was done using a standard protocol recording all relevant information for radiation exposure, i.e. radiation protection devices and tools. This study shows the preliminary results obtained for this measurement campaign. For diagnostic purposes, the two most-used radionuclides were considered: (99m)Tc and (18)F. For therapeutic treatments, Zevalin(®) and DOTATOC (both labelled with (90)Y) were chosen. Large variations of doses were observed across the hands depending on different parameters. Furthermore, this study highlights the importance of the positioning of the extremity dosemeter for a correct estimate of the maximum skin doses.

Contribución al diseño de GNSS-SDR, un receptor GNSS de código abierto

[ANGLÈS] This project introduces GNSS-SDR, an open source Global Navigation Satellite System software-defined receiver. The lack of reconfigurability of current commercial-of-the-shelf receivers and the advent of new radionavigation signals and systems make software receivers an appealing approach to design new architectures and signal processing algorithms. With the aim of exploring the full potential of this forthcoming scenario with a plurality of new signal structures and frequency bands available for positioning, this paper describes the software architecture design and provides details about its implementation, targeting a multiband, multisystem GNSS receiver. The result is a testbed for GNSS signal processing that allows any kind of customization, including interchangeability of signal sources, signal processing algorithms, interoperability with other systems, output formats, and the offering of interfaces to all the intermediate signals, parameters and variables. The source code release under the GNU General Public License (GPL) secures practical usability, inspection, and continuous improvement by the research community, allowing the discussion based on tangible code and the analysis of results obtained with real signals. The source code is complemented by a development ecosystem, consisting of a website (http://gnss-sdr.org), as well as a revision control system, instructions for users and developers, and communication tools. The project shows in detail the design of the initial blocks of the Signal Processing Plane of the receiver: signal conditioner, the acquisition block and the receiver channel, the project also extends the functionality of the acquisition and tracking modules of the GNSS-SDR receiver to track the new Galileo E1 signals available. Each section provides a theoretical analysis, implementation details of each block and subsequent testing to confirm the calculations with both synthetically generated signals and with real signals from satellites in space.

Décubitus ventral et syndrome de détresse respiratoire aiguë de l'adulte: de la théorie à la pratique [Prone position mechanical ventilation in the Acute Respiratory Distress syndrome: theoretical and practical considerations].

Mortality of the acute respiratory distress syndrome (ARDS) remains extremely high and only few evidence-based specific treatments are currently available. Protective mechanical ventilation has emerged as the comer stone of the management of ARDS to avoid the occurrence of ventilation-induced lung injuries (VILI). Mechanical ventilation in the prone position has often been considered as a rescue therapy reserved to refractory hypoxemia. Since the publication of the PROSEVA study in 2013, early prone positioning for mechanical ventilation should be recommended to improve survival of patients with severe ARDS. In this article, both the theoretical and practical aspects of mechanical ventilation in prone position are reviewed.