Visualization of RNA polymerase II ternary transcription complexes formed in vitro on a Xenopus laevis vitellogenin gene.

Stable ternary transcription complexes assembled in vitro, using a HeLa whole-cell extract, have been isolated and visualized by electron microscopy. The formation of these stable complexes on the DNA fragment used as template, the 5' end region of the Xenopus laevis vitellogenin gene B2, depends on factors present in the whole-cell extract, RNA polymerase II and at least two nucleotides. Interestingly, bending in the DNA fragment was frequently observed at the binding site of RNA polymerase II. Dinucleotides that can prime initiation within a short sequence of approximately 10 contiguous nucleotides centered around the initiation site used in vivo, also favour the formation of stable complexes. In addition, pre-initiation complexes were isolated and it was shown that factors in the extract involved in their formation are more abundant than the RNA polymerase II molecules available for binding. The possible implication of this observation relative to the in vivo situation is discussed.

Protein-protein interactions between adenovirus DNA polymerase and nuclear factor I mediate formation of the DNA replication preinitiation complex.

The in vitro adenovirus (Ad) DNA replication system provides an assay to study the interaction of viral and host replication proteins with the DNA template in the formation of the preinitiation complex. This initiation system requires in addition to the origin DNA sequences 1) Ad DNA polymerase (Pol), 2) Ad preterminal protein (pTP), the covalent acceptor for protein-primed DNA replication, and 3) nuclear factor I (NFI), a host cell protein identical to the CCAAT box-binding transcription factor. The interactions of these proteins were studied by coimmunoprecipitation and Ad origin DNA binding assays. The Ad Pol can bind to origin sequences only in the presence of another protein which can be either pTP or NFI. While NFI alone can bind to its origin recognition sequence, pTP does not specifically recognize DNA unless Ad Pol is present. Thus, protein-protein interactions are necessary for the targetting of either Ad Pol or pTP to the preinitiation complex. DNA footprinting demonstrated that the Ad DNA site recognized by the pTP.Pol complex was within the first 18 bases at the end of the template which constitutes the minimal origin of replication. Mutagenesis studies have defined the Ad Pol interaction site on NFI between amino acids 68-150, which overlaps the DNA binding and replication activation domain of this factor. A putative zinc finger on the Ad Pol has been mutated to a product that fails to bind the Ad origin sequences but still interacts with pTP. These results indicate that both protein-protein and protein-DNA interactions mediate specific recognition of the replication origin by Ad DNA polymerase.

Cooperative kinking at distant sites in mechanically stressed DNA.

In cells, DNA is routinely subjected to significant levels of bending and twisting. In some cases, such as under physiological levels of supercoiling, DNA can be so highly strained, that it transitions into non-canonical structural conformations that are capable of relieving mechanical stress within the template. DNA minicircles offer a robust model system to study stress-induced DNA structures. Using DNA minicircles on the order of 100 bp in size, we have been able to control the bending and torsional stresses within a looped DNA construct. Through a combination of cryo-EM image reconstructions, Bal31 sensitivity assays and Brownian dynamics simulations, we have been able to analyze the effects of biologically relevant underwinding-induced kinks in DNA on the overall shape of DNA minicircles. Our results indicate that strongly underwound DNA minicircles, which mimic the physical behavior of small regulatory DNA loops, minimize their free energy by undergoing sequential, cooperative kinking at two sites that are located about 180° apart along the periphery of the minicircle. This novel form of structural cooperativity in DNA demonstrates that bending strain can localize hyperflexible kinks within the DNA template, which in turn reduces the energetic cost to tightly loop DNA.

« Les enjeux de l'analyse contrastive dans l'Union Soviétique des années 1920 »

http://w3.u-grenoble3.fr/ellug/index.html/fileadmin/template/ellug/Telechargements/tdm_grammaire_et_lexique.pdf

Nonspecific PCR amplification by high-fidelity polymerases: implications for next-generation sequencing of AFLP markers.

High-fidelity 'proofreading' polymerases are often used in library construction for next-generation sequencing projects, in an effort to minimize errors in the resulting sequence data. The increased template fidelity of these polymerases can come at the cost of reduced template specificity, and library preparation methods based on the AFLP technique may be particularly susceptible. Here, we compare AFLP profiles generated with standard Taq and two versions of a high-fidelity polymerase. We find that Taq produces fewer and brighter peaks than high-fidelity polymerase, suggesting that Taq performs better at selectively amplifying templates that exactly match the primer sequences. Because the higher accuracy of proofreading polymerases remains important for sequencing applications, we suggest that it may be more effective to use alternative library preparation methods.

First aid in burn injuries with counterintuitive warm water improves outcome

Objective: Cooling is considered a panacea in burn injury. However, burn injuries are characterized by an ischemic zone prone to progression, a phenomenon that can substantially increase morbidity. Cold-induced vasoconstriction potentially aggravates ischemia and promotes progression. Therefore we compared the effect of warm (37°C) and cold (17°C) water on burn progression. Methods: The comb burn model creates 4 rectangular burned surfaces separated by 3 unburned interspaces that become necrotic if untreated. After heating in boiling water the template was applied for 60 seconds on 24 Wistar rats randomized into 3 groups: no treatment (CON); treatment for 20 minutes with cold water (17°C: CW) or warm water (37°C: WW). Burn progression in surface (planimetry) and Departmenth (histology), as well as microcirculatory perfusion (laser Doppler flowmetry) were assessed after 1h, as well as 1, 4, and 7 days. Results: Both CW and WW delayed burn progression without reducing the final burn Departmenth (deep dermis). In contrast, only WW but not CW increased dermal perfusion (81 ± 2% (WW) vs. 62 ± 2% (CW) and 63 ± 1% (CON), p< 0·05) already 1 hour after burn induction. The difference observed after one hour led to a complete flow recovery during the observation period and translated into increased interspace survival, respectively less necrosis with WW(65 ± 4% vs. 81 + 4% (CW) and 91 ± 2% (CON), p< 0·05) after 7 days. Conclusions: Application of warm water significantly improved dermal perfusion, increased interspace survival, and delayed burn progression.However it didn't alter the ultimate burn Departmenth of the actually burned area. Therefore, warm water can create a therapeutic window for targeted nonsurgical treatment of burn progression.

Utilização do dióxido de carbono supercrítico na concentração de tocoferóis do destilado desodorizado do óleo de soja

O objetivo deste trabalho foi examinar a possibilidade do uso do dióxido de carbono supercrítico na extração e concentração de tocoferóis a partir do destilado desodorizado do óleo de soja. Trata-se da combinação seqüencial de duas extrações: pré-extração à temperatura de 80°C e pressão de 76 bars para remoção de substâncias interferentes e extração dos tocoferóis a 50°C e 197 bars. O coletor utilizado foi o octadesil-sílica, lavado com acetonitrila na pré-extração e hexano na extração, e a análise dos extratos obtidos foi feita por cromatografia de fase gasosa em coluna capilar. O resultado mostrou que o desodorizado do óleo de soja, contendo inicialmente 9,2% de tocoferóis totais, pôde ser concentrado para 40,6%. Sob as condições otimizadas, a extração pode ser realizada em 37 minutos.

Vulnerabilities of Fingerprint Reader to Fake Fingerprint Attacks

The purpose of this research is to assess the vulnerabilities of a high resolution fingerprint sensor when confronted with fake fingerprints. The study has not been focused on the decision outcome of the biometric device, but essentially on the scores obtained following the comparison between a query (genuine or fake) and a template using an AFIS system. To do this, fake fingerprints of 12 subjects have been produced with and without their cooperation. These fake fingerprints have been used alongside with real fingers. The study led to three major observations: First, genuine fingerprints produced scores higher than fake fingers (translating a closer proximity) and this tendency is observed considering each subject separately. Second, scores are however not sufficient as a single measure to differentiate these samples (fake from genuine) given the variation due to the donors themselves. That explains why fingerprint readers without vitality detection can be fooled. Third, production methods and subjects greatly influence the scores obtained for fake fingerprints.

Tolerância ao alumínio em porta-enxertos somaclonais de macieira cultivados em solução nutritiva

A macieira (Malus sp.) cultivada em solos ácidos com pH inferior a 5,0 pode apresentar distúrbios nutricionais e inibição de crescimento causados por níveis tóxicos de Al que podem afetar a absorção do Ca, o qual apresenta estreita correlação com a produção de frutos de melhor qualidade. O objetivo deste trabalho foi verificar a tolerância ao Al em 18 somaclones e três cultivares porta-enxertos provenientes da seleção in vitro. As plântulas foram cultivadas durante 45 dias em copos de plástico de 300 mL contendo solução nutritiva e utilizando-se sílica lavada como substrato. Os valores obtidos foram submetidos à análise multivariada pelos componentes principais e agrupamento hierárquico baseado no método de Ward, o qual revelou ser mais adequado na classificação dos clones quanto à tolerância ao Al do que os testes de médias. O porcentual de redução na matéria seca da parte aérea e no número de folhas foram características que melhor possibilitaram a discriminação dos clones. Os clones foram classificados em três grupos: bem sensível ou não tolerante, moderadamente tolerante e tolerante.

Iowa School Safety Guide, April 6, 2012

This document was developed for the schools of Iowa to use as a template to enhance current school safety programs; the creation of this document was a partnered effort at the state level between the aforementioned agencies. The purpose of this document is to give school districts and individual schools a planning resource to use when creating their school safety plans. Ultimately, schools can decide how much of this document they would like to incorporate into their current plan. The original document was created by the Minnesota Department of Homeland Security, and its use was granted to Iowa Homeland Security in 2011. Iowa pulled together a panel of experts to make this document specific to Iowa’s schools, and laws. It’s important to note the partnership created by this document is intended to continue through information sharing in relation to critical assets, infrastructure protection, and school safety. Iowa Homeland Security is a representative in the Iowa Department of Public Safety, Division of Intelligence Fusion Center. This partnership allows for streamlined information sharing to the critical infrastructure owner/operators across the state. The current plan for information sharing is through the Iowa Homeland Security and Emergency Management, Threat information and Infrastructure Protection Program (TIIPP) to the Iowa Department of Education for processing and dissemination statewide. Depending on the type of information being released it could be specific to a school, district or the education sector statewide.

A knowledge based signal processing approach to tonic identification in indian classical music

In this paper, we describe several techniques for detecting tonic pitch value in Indian classical music. In Indian music, the raga is the basic melodic framework and it is built on the tonic. Tonic detection is therefore fundamental for any melodic analysis in Indian classical music. This workexplores detection of tonic by processing the pitch histograms of Indian classic music. Processing of pitch histograms using group delay functions and its ability to amplify certain traits of Indian music in the pitch histogram, is discussed. Three different strategies to detect tonic, namely, the concert method, the template matching and segmented histogram method are proposed. The concert method exploits the fact that the tonic is constant over a piece/concert.templatematchingmethod and segmented histogrammethodsuse the properties: (i) the tonic is always present in the background, (ii) some notes are less inflected and dominant, to detect the tonic of individual pieces. All the three methods yield good results for Carnatic music (90−100% accuracy), while for Hindustanimusic, the templatemethod works best, provided the v¯adi samv¯adi notes for a given piece are known (85%).

Enhancing centralized enforcement of EU law : Pandora's toolbox ?

State compliance with EU Law is crucial to the very existence of the Union. Traditionally, it has been secured through a combination of strong "private" and of weak "centralized" enforcement. However, this arrangement is no longer perceived to be sufficient. By endowing the Union with new tools vis-à-vis its Member States - penalties, conditionality, and the like - current reforms try to complement symbolic sanctioning with real "consequences". The goal is to reinforce the authority of EU Law. In this article, we question whether the new toolbox is fit for the purpose, or whether it risks to produce adverse effects which might even go as far as upsetting the Union's constitutional template.

Anàlisi i quantificació de les accions i el temps de joc en l'hoquei sobre patins en equips de l'OK Lliga

Quantificar les variables temporal, les accions tècniques i els desplaçaments dels jugador al llarg de la competició proporciona una informació excel·lent per deduir les càrregues físiques i fisiològiques a les que estan sotmesos. Per aquest estudi es van visualitzar 4 partits de la Copa del Rei 2012. Es va enregistrà sistemàticament les dades en fulls de registre. Les principals dades analitzades van ser: temps total, temps real, temps de pausa, temps d’acció i relació entre el temps d’acció i el temps de pausa (densitat de treball). També es va analitzar: les incidències reglamentaries, les accions tècniques del jugador i els desplaçaments del jugador. El 45% de les accions es van produir entre 0 i 30s, i el promig d’interval de participació va ser 19:29s ±2:09. Més del 50% de les pauses es van produir entre 11s i 30s, i el promig d’interval de pausa va ser de 24:57s. Aquest va determinar que la densitat de treball era 1:1.3. Aproximadament el 70% de les incidències reglamentàries van ser faltes d’equip I tècniques. Van haver-hi 56 conduccions, 10 driblins, 12 remats, (3 xuts, 2 punxades/escopides, 5 remats de primeres i 2 arrossegades) i 80 passades. Finalment, en un partit el 42,23% dels desplaçaments van ser a baixa intensitat, el 27,02% van ser a intensitat mitjana i el 30,75% van ser a alta intensitat. A més a més, el 81,90% dels desplaçaments a alta intensitat van durar de 0s a 5s, el 54,56% a intensitat mitjana van durar de 6 a 10 s. I el 49,32% a intensitat baixa també van durar 6s a 10s. L’hoquei sobre patins és un esport en el que hi predominen els esforços intermitents a gran intensitat i de curta durada amb una important sol·licitació de la via anaeròbica alàctica. Analitzar la dimensió temporal, les accions tècniques i els desplaçaments dels jugador durant la competició permet estimar les exigències físiques i requeriments energètics, i aquest faciliten l’elaboració d’entrenaments més específics.

A review of atlas-based segmentation for magnetic resonance brain images.

Normal and abnormal brains can be segmented by registering the target image with an atlas. Here, an atlas is defined as the combination of an intensity image (template) and its segmented image (the atlas labels). After registering the atlas template and the target image, the atlas labels are propagated to the target image. We define this process as atlas-based segmentation. In recent years, researchers have investigated registration algorithms to match atlases to query subjects and also strategies for atlas construction. In this paper we present a review of the automated approaches for atlas-based segmentation of magnetic resonance brain images. We aim to point out the strengths and weaknesses of atlas-based methods and suggest new research directions. We use two different criteria to present the methods. First, we refer to the algorithms according to their atlas-based strategy: label propagation, multi-atlas methods, and probabilistic techniques. Subsequently, we classify the methods according to their medical target: the brain and its internal structures, tissue segmentation in healthy subjects, tissue segmentation in fetus, neonates and elderly subjects, and segmentation of damaged brains. A quantitative comparison of the results reported in the literature is also presented.

A high-efficiency HeLa cell nuclear transcription extract.

A HeLa cell nuclear transcription extract that is approximately 20 times more efficient than standard HeLa cell transcription extracts was developed. Transcription of the strong adenovirus II major late promoter by this extract results in the synthesis of 1.5-4 molecules of product RNA per molecule of template, indicating that the extract is capable of multiple rounds of initiation. Standard HeLa cell nuclear extracts transcribe closed circular and linear adenovirus major late promoter templates with equal efficiency. In contrast, the new extract exhibits an increase of approximately twofold on transcription of a closed circular, as opposed to a linear, major late promoter template.