Microscale mesoarrays created by dip-pen nanolithography for screening of protein-protein interactions

Using microarrays to probe protein-protein interactions is becoming increasingly attractive due to their compatibility with highly sensitive detection techniques, selectivity of interaction, robustness and capacity for examining multiple proteins simultaneously. The major drawback to using this approach is the relatively large volumes and high concentrations necessary. Reducing the protein array spot size should allow for smaller volumes and lower concentrations to be used as well as opening the way for combination with more sensitive detection technologies. Dip-Pen Nanolithography (DPN) is a recently developed technique for structure creation on the nano to microscale with the capacity to create biological architectures. Here we describe the creation of miniaturised microarrays, 'mesoarrays', using DPN with protein spots 400× smaller by area compared to conventional microarrays. The mesoarrays were then used to probe the ERK2-KSR binding event of the Ras/Raf/MEK/ERK signalling pathway at a physical scale below that previously reported. Whilst the overall assay efficiency was determined to be low, the mesoarrays could detect KSR binding to ERK2 repeatedly and with low non-specific binding. This study serves as a first step towards an approach that can be used for analysis of proteins at a concentration level comparable to that found in the cellular environment.

The extreme right in France:from Pétain to Le Pen

The Front National has for some years been France's third political party and the most notable far-right force in Europe; its leader, Jean-Marie Le Pen, contested the 2002 presidential election run-off with 5.5 million votes. What do Le Pen and the FN represent? What are their historical roots, their values and their policies? Who votes for them and why? And what has been their impact on the political agenda in France? Adopting an essentially chronological approach, the book traces the political lineage of Le Pen and the FN through key figures, movements and events on the French extreme right from the Vichy regime to the present, providing a detailed historical perspective for understanding the FN today. Part I provides a historical study of the extreme right in France since 1940, examining • the Vichy regime, collaboration and ‘collaborationism’, • the aftermath of Liberation and the post-war extreme right, • the Poujadist movement and the politics of populism, • the Algerian War as a catalyst for change, • the ‘Nouvelle Droite’ and the search for doctrinal renewal, • old and new forms of extreme-right ideology and activism. Part II undertakes a comprehensive study of the FN, analysing • the party’s early development and electoral rise, • its evolving programme and strategy, • the factors underlying its popular appeal, • the geography and composition of its electorate, • its exercise of local power and regional influence, • and its defining impact on the national political agenda. The FN, it is argued, represents both the latest manifestation of a long tradition of authoritarian nationalism and a complex new phenomenon within the changing social and political dynamics of contemporary France.

Support for Le Pen in France:two elections in trompe l’oeil

There is a comforting consensus among political commentators that the 2007 presidential election marked the end of Jean-Marie Le Pen as a force in French politics. The shock election of the Front National leader to the presidential run-off in 2002, by contrast, is explained as a surge in the Le Pen vote specific to the prevailing electoral conditions. This article challenges that interpretation of both elections. It shows that, despite Le Pen’s unforeseen success in 2002, there was no surge of support for him, and that despite Le Pen’s supposed collapse in 2007, he won close to 4 million votes while popular agreement with his ideas rose to its highest recorded level. The article concludes that Le Pen remains a powerful presence in French politics and that his supporters continue to constitute a large and highly influential constituency.

Marine Le Pen and the 'New' FN:a change of style or of substance?

The electoral challenge of the far right is an enduringly problematic feature of contemporary French politics. In the first rounds of the 2012 presidential and parliamentary elections, the Front National (FN) under new leader Marine Le Pen attracted a combined total of ten million votes, bringing its ultra-nationalist policies to the centre of national political debate. This article examines the FN's impact on these elections and its implications for French politics. Drawing on official FN programmes, detailed election results and a range of opinion polling data, it assesses the strength of support for Le Pen and her party and seeks to explain their electoral appeal. In particular, it subjects to analysis the claim that the new leader has ‘de-demonised’ the FN, transforming it from perennial outsider to normal participant in mainstream French politics; and it reflects on the strategic dilemma posed for the centre-right by this newly invigorated far-right challenge.

Seismic reflection data, calcium/iron ratios, stable istopes, and magnetic susceptibility of sediment cores from the Pen Duick Escarpment, Gulf of Cadiz

Here we present a case study of three cold-water coral mounds in a juvenile growth stage on top of the Pen Duick Escarpment in the Gulf of Cadiz; Alpha, Beta and Gamma mounds. Although cold-water corals are a common feature on the adjacent cliffs, mud volcanoes and open slope, no actual living cold-water coral has been observed. This multidisciplinary and integrated study comprises geophysical, sedimentological and (bio)geochemical data and aims to present a holistic view on the interaction of both environmental and geological drivers in cold-water coral mound development in the Gulf of Cadiz. Coring data evidences (past or present) methane seepage near the Pen Duick Escarpment. Several sources and pathways are proposed, among which a stratigraphic migration through uplifted Miocene series underneath the escarpment. The dominant morphology of the escarpment has influenced the local hydrodynamics within the course of the Pliocene, as documented by the emplacement of a sediment drift. Predominantly during post-Middle Pleistocene glacial episodes, favourable conditions were present for mound growth. An additional advantage for mound formation near the top of Pen Duick Escarpment is presented by seepage-related carbonate crusts which might have offered a suitable substrate for coral settling. The spatially and temporally variable character and burial stage of the observed open reef frameworks, formed by cold-water coral rubble, provides a possible model for the transition from cold-water coral reef patches towards juvenile mound. These rubble "graveyards" not only act as sediment trap but also as micro-habitat for a wide range of organisms. The presence of a fluctuating Sulphate-Methane Transition Zone has an important effect on early diagenetic processes, affecting both geochemical and physical characteristics, transforming the buried reef into a solid mound. Nevertheless, the responsible seepage fluxes seem to be locally variable. As such, the origin and evolution of the cold-water coral mounds on top of the Pen Duick Escarpment is, probably more than any other NE Atlantic cold-water coral mound province, located on the crossroads of environmental (hydrodynamic) and geological (seepage) pathways.

Implementing the European quality assurance in vocational education and training (EQAVET) at national level: Some insights from the PEN Leonardo project

EQAVET, the quality assurance tool in vocational and educational training, was developed in response to the need for a supply of a trained workforce for labour market needs. Implementation of EQAVET at national level, however, remains a challenge. The research reported here focused on the implementation of QA processes by VET providers in 4 countries: Malta, Italy, Turkey, and Sweden. Data was collected through a questionnaire with 62 VET providers. Responses showed that there is an overall commitment to quality. There is, however, little knowledge of EQAVET across the countries, with the exception of Malta. None the less, all VET providers have implemented some aspects of EQAVET, even if not always intentionally. The situation is, however, far from EQAVET being fully implemented. Reflections are made on whether the EQAVET model specifically or qualification assurance principles assurances should be promoted across Europe. (DIPF/Orig.)

The Sword and the Pen: Life Writings by Militant-Authors of the Việt Minh and Front de Libération Nationale (FLN)

This dissertation examines four life writings by militant-authors of the Việt Minh and Front de la Libération Nationale (FLN): Ngô Văn Chiêu’s Journal d’un combattant Viet-Minh (1955), Đặng Văn Việt’s De la RC 4 à la N 4: la campagne des frontières (2000), Si Azzedine’s On nous appelait fellaghas (1976), and Saadi Yacef’s two-volume La Bataille d’Alger (2002). In describing the Vietnamese and Algerian Revolutions through the perspectives of combatants who participated in their respective countries’ national liberation struggles, the texts reveal that four key factors motivated the militants and led them to believe that independence was historically inevitable: (1) a philosophical, political, and ideological framework, (2) the support of multiple segments of the local population, (3) the effective use of guerrilla and psychological warfare, and (4) military, moral, and political assistance provided by international allies. By fighting for the independence of their countries and documenting their revolutionary experiences, the four militant-authors leave their mark on the world using both the sword and the pen.

Formación en el oficio artesanal del trabajo en totumo como estrategia para contribuir a la generación de actividades productivas en Peñoncito, municipio de El Peñón, Sur de Bolívar.

157 hojas : ilustraciones, fotografías.

Effect Of Pain Chronification And Chronic Pain On An Endogenous Pain Modulation Circuit In Rats.

We tested the hypothesis that chronic pain development (pain chronification) and ongoing chronic pain (chronic pain) reduce the activity and induce plastic changes in an endogenous analgesia circuit, the ascending nociceptive control. An important mechanism mediating this form of endogenous analgesia, referred to as capsaicin-induced analgesia, is its dependence on nucleus accumbens μ-opioid receptor mechanisms. Therefore, we also investigated whether pain chronification and chronic pain alter the requirement for nucleus accumbens μ-opioid receptor mechanisms in capsaicin-induced analgesia. We used an animal model of pain chronification in which daily subcutaneous prostaglandin E2 (PGE2) injections into the rat's hind paw for 14 days, referred to as the induction period of persistent hyperalgesia, induce a long-lasting state of nociceptor sensitization referred to as the maintenance period of persistent hyperalgesia, that lasts for at least 30 days following the cessation of the PGE2 treatment. The nociceptor hypersensitivity was measured by the shortening of the time interval for the animal to respond to a mechanical stimulation of the hind paw. We found a significant reduction in the duration of capsaicin-induced analgesia during the induction and maintenance period of persistent mechanical hyperalgesia. Intra-accumbens injection of the μ-opioid receptor selective antagonist Cys(2),Tyr(3),Orn(5),Pen(7)amide (CTOP) 10 min before the subcutaneous injection of capsaicin into the rat's fore paw blocked capsaicin-induced analgesia. Taken together, these findings indicate that pain chronification and chronic pain reduce the duration of capsaicin-induced analgesia, without affecting its dependence on nucleus accumbens μ-opioid receptor mechanisms. The attenuation of endogenous analgesia during pain chronification and chronic pain suggests that endogenous pain circuits play an important role in the development and maintenance of chronic pain.

Validity and reliability of methods for the detection of secondary caries around amalgam restorations in primary teeth

Secondary caries has been reported as the main reason for restoration replacement. The aim of this in vitro study was to evaluate the performance of different methods - visual inspection, laser fluorescence (DIAGNOdent), radiography and tactile examination - for secondary caries detection in primary molars restored with amalgam. Fifty-four primary molars were photographed and 73 suspect sites adjacent to amalgam restorations were selected. Two examiners evaluated independently these sites using all methods. Agreement between examiners was assessed by the Kappa test. To validate the methods, a caries-detector dye was used after restoration removal. The best cut-off points for the sample were found by a Receiver Operator Characteristic (ROC) analysis, and the area under the ROC curve (Az), and the sensitivity, specificity and accuracy of the methods were calculated for enamel (D2) and dentine (D3) thresholds. These parameters were found for each method and then compared by the McNemar test. The tactile examination and visual inspection presented the highest inter-examiner agreement for the D2 and D3 thresholds, respectively. The visual inspection also showed better performance than the other methods for both thresholds (Az = 0.861 and Az = 0.841, respectively). In conclusion, the visual inspection presented the best performance for detecting enamel and dentin secondary caries in primary teeth restored with amalgam.

A novel disposable electrochemical microcell: construction and characterization

An alternative technique for the fabrication of disposable electrochemical microcells containing working, reference and auxiliary electrodes on a single device is reported. The procedure is based on thermal-transfer of toner masks onto CD-R (recordable compact discs) gold surfaces to define the layout of the electrodes (contour). In a subsequent step, the layout is manually painted with a permanent marker pen. The unprotected gold surface is conveniently etched (chemical corrosion) and the ink is then easily removed with ethanol, generating gold surfaces without contamination. The final and reproducible area of the electrodes is defined by heat transference of a second toner mask. Silver epoxy is deposited on one of the gold bands which is the satisfactorily used as reference electrode. These microcells were electrochemically characterized by cyclic, linear, and square wave voltammetry, and several electroactive species were used as model systems. The area reproducibility of the electrodes for different microcells was studied and a relative standard deviation better than 1,0% (n = 10) was obtained. Disposable electrochemical microcells were successfully used in analysis of liquid samples with volumes lower than 200 µL and good stability and reproducibility (RSD less than 2.0%) were achieved. These microcells were also evaluated for quantification of paracetamol and dipyrone in pharmaceutical formulations.

New methodology to assess activity status of occlusal caries in primary teeth using laser fluorescence device

An in vivo study was conducted to verify the ability of laser fluorescence (LF) to assess the activity status of occlusal caries in primary teeth, using different air-drying times. Occlusal sites (707) were examined using LF (DIAGNOdent) after air-drying for 3 s and 15 s, and the difference between readings (DIF15 s-3 s) was calculated. For concurrent validation of LF, visual criteria-Nyvad (NY) and Lesion Activity Assessment associated with the International Caries Detection and Assessment System (LAA-ICDAS)-were the reference standards for lesion activity. Histological exam using a pH-indicator dye (0.1% methyl red) was performed in 46 exfoliated/extracted teeth for criterion validation. LF readings and DIF15 s-3 s were compared using Kruskall-Wallis and Mann-Whitney tests. Receiver operating characteristic analyses were performed and validity parameters calculated, considering the caries activity assessment. Using NY, active lesions (3 s: 30.0 +/- 29.3; 15 s: 34.2 +/- 30.6) presented higher LF readings than inactive lesions (3 s: 17.0 +/- 16.3; 15 s: 19.2 +/- 17.3; p <0.05), different from LAA-ICDAS. Active cavitated caries resulted in higher LF readings (3 s: 50.3 +/- 3.5; 15 s: 54.7 +/- 30.2) than inactive cavitated caries (3 s: 19.9 +/- 16.3; 15 s: 22.8 +/- 16.8). Therefore, LF can distinguish cavitated active and inactive lesions classified by NY, but not by LAA-ICDAS; however, this difference might be related to the visual system rather than to LF. The air-drying time could be an alternative to improve the caries activity assessment; however, longer air-drying time is suggested to be tested subsequently. (C) 2010 Society of Photo-Optical Instrumentation Engineers. [DOI: 10.1117/1.3463007]

Programmable diagnostic devices made from paper and tape

This paper describes three-dimensional microfluidic paper-based analytical devices (3-D mu PADs) that can be programmed (postfabrication) by the user to generate multiple patterns of flow through them. These devices are programmed by pressing single-use 'on' buttons, using a stylus or a ballpoint pen. Pressing a button closes a small space (gap) between two vertically aligned microfluidic channels, and allows fluids to wick from one channel to the other. These devices are simple to fabricate, and are made entirely out of paper and double-sided adhesive tape. Programmable devices expand the capabilities of mu PADs and provide a simple method for controlling the movement of fluids in paper-based channels. They are the conceptual equivalent of field-programmable gate arrays (FPGAs) widely used in electronics.