Mutational analysis of NM23-H2/NDP kinase identifies the structural domains critical to recognition of a c-myc regulatory element.

NM23-H2, a presumed regulator of tumor metastasis in humans, is a hexameric protein with both enzymatic (NDP kinase) and regulatory (transcriptional activation) activity. While the structure and catalytic mechanisms have been well characterized, the mode of DNA binding is not known. We examined this latter function in a site-directed mutational study and identified residues and domains essential for the recognition of a c-myc regulatory sequence. Three amino acids, Arg-34, Asn-69, and Lys-135, were found among 30 possibilities to be critical for DNA binding. Two of these, Asn-69 and Lys-135, are not conserved between NM23 variants differing in DNA-binding potential, suggesting that DNA recognition resides partly in nonconserved amino acids. All three DNA-binding defective mutant proteins are active enzymatically and appear to be stable hexamers, suggesting that they perform at the level of DNA recognition and that separate functional domains exist for enzyme catalysis and DNA binding. In the context of the known crystal structure of NM23-H2, the DNA-binding residues are located within distinct structural motifs in the monomer, which are exposed to the surface near the 2-fold axis of adjacent subunits in the hexamer. These findings are explained by a model in which NM23-H2 binds DNA with a combinatorial surface consisting of the "outer" face of the dimer. Chemical crosslinking data support a dimeric DNA-binding mode by NM23-H2.

Self-recognition in primates: phylogeny and the salience of species-typical features.

Self-recognition has been explored in nonlinguistic organisms by recording whether individuals touch a dye-marked area on visually inaccessible parts of their face while looking in a mirror or inspect parts of their body while using the mirror's reflection. Only chimpanzees, gorillas, orangutans, and humans over the age of approximately 2 years consistently evidence self-directed mirror-guided behavior without experimenter training. To evaluate the inferred phylogenetic gap between hominoids and other animals, a modified dye-mark test was conducted with cotton-top tamarins (Saguinus oedipus), a New World monkey species. The white hair on the tamarins' head was color-dyed, thereby significantly altering a visually distinctive species-typical feature. Only individuals with dyed hair and prior mirror exposure touched their head while looking in the mirror. They looked longer in the mirror than controls, and some individuals used the mirror to observe visually inaccessible body parts. Prior failures to pass the mirror test may have been due to methodological problems, rather than to phylogenetic differences in the capacity for self-recognition. Specifically, an individual's sensitivity to experimentally modified parts of its body may depend crucially on the relative saliency of the modified part (e.g., face versus hair). Moreover, and in contrast to previous claims, we suggest that the mirror test may not be sufficient for assessing the concept of self or mental state attribution in nonlinguistic organisms.

Molecular approaches to smart materials via interface recognition and conformationally switchable cavitands

In this thesis the molecular level design of functional materials and systems is reported. In the first part, tetraphosphonate cavitand (Tiiii) recognition properties towards amino acids are studied both in the solid state, through single crystal X-ray diffraction, and in solution, via NMR and ITC experiments. The complexation ability of these supramolecular receptors is then applied to the detection of biologically remarkable N-methylated amino acids and peptides using complex dynamic emulsions-based sensing platforms. In the second part, a general supramolecular approach for surface decoration with single-molecule magnets (SMMs) is presented. The self-assembly of SMMs is achieved through the formation of a multiple hydrogen bonds architecture (UPy-NaPy complexation). Finally we explore the possibility to impart auxetic behavior to polymeric material through the introduction of conformationally switchable monomers, namely tetraquinoxaline cavitands (QxCav). Their interconversion from a closed vase conformation to an extended kite form is studied first in solution, then in polymeric matrixes via pH and tensile stimuli by UV-Vis spectroscopy.

Biochemical characterization and DNA binding properties of the MocR family member GabR, a PLP-dependent transcription factor

GabR è un fattore di trascrizione chimerico appartenente alla famiglia dei MocR/GabR, costituito da un dominio N-terminale elica-giro-elica di legame al DNA e un dominio effettore e/o di oligomerizzazione al C-terminale. I due domini sono connessi da un linker flessibile di 29 aminoacidi. Il dominio C-terminale è strutturalmente omologo agli enzimi aminotransferasici fold-type I, i quali, utilizzando il piridossal-5’-fosfato (PLP) come cofattore, sono direttamente coinvolti nel metabolismo degli aminoacidi. L’interazione contemporanea di PLP e acido γ-aminobutirrico (GABA) a GabR fa sì che questa promuova la trascrizione di due geni, gabT e gabD, implicati nel metabolismo del GABA. GabR cristallizza come un omodimero con una configurazione testa-coda. Il legame con la regione promotrice gabTD avviene attraverso il riconoscimento specifico di due sequenze dirette e ripetute (ATACCA), separate da uno spacer di 34 bp. In questo studio sono state indagate le proprietà biochimiche, strutturali e di legame al DNA della proteina GabR di Bacillus subtilis. L’analisi spettroscopica dimostra che GabR interagisce con il PLP formando l’aldimina interna, mentre in presenza di GABA si ottiene l’aldimina esterna. L’interazione fra il promotore gabTD e le forme holo e apo di GabR è stata monitorata mediante Microscopia a Forza atomica (AFM). In queste due condizioni di legame è stata stimata una Kd di circa 40 ηM. La presenza di GABA invece, determinava un incremento di circa due volte della Kd, variazioni strutturali nei complessi GabR-DNA e una riduzione del compattamento del DNA alla proteina, indipendentemente dalla sequenza del promotore in esame. Al fine di valutare il ruolo delle caratteristiche topologiche del promotore, sono state inserite cinque e dieci bp all’interno della regione spacer che separa le due sequenze ripetute dirette riconosciute da GabR. I significativi cambiamenti topologici riscontrati nel frammento aggiunto di cinque bp si riflettono anche sulla forte riduzione dell’affinità di legame verso la proteina. Al contrario, l’inserzione di 10 bp provoca solamente l’allontanamento delle sequenze ripetute dirette. L’assenza quindi di cambiamenti significativi nella topologia di questo promotore fa sì che l’affinità di legame per GabR rimanga pressoché inalterata rispetto al promotore non mutato. L’analisi del potenziale elettrostatico superficiale di GabR mostra la presenza di una fascia carica positivamente che si estende lungo un’intera faccia della proteina. Per verificare l’importanza di questa caratteristica di GabR nel meccanismo di interazione al DNA, sono stati preparati ed indagati i mutanti R129Q e K362-366Q, in cui la carica positiva superficiale risultava indebolita. L’affinità di legame dei mutanti di GabR per il DNA era inferiore rispetto alla proteina non mutata, in particolar modo nel mutante K362-366Q. Le evidenze acquisite suggeriscono che la curvatura intrinseca del promotore ed il corretto orientamento delle sequenze sulla doppia elica, più della distanza che le separa, siano critici per sostenere l’interazione con GabR. Oltre a questo, la superficie positiva di GabR è richiesta per accomodare la curvatura del DNA sul corpo della proteina. Alla luce di questo, l’interazione GabR-gabTD è un esempio di come il riconoscimento specifico di sequenze, la topologia del DNA e le caratteristiche strutturali della proteina siano contemporaneamente necessarie per sostenere un’interazione proteina-DNA stabile.

Grammar specification for the recognition of temporal expressions

This paper tells about the recognition of temporal expressions and the resolution of their temporal reference. A proposal of the units we have used to face up this tasks over a restricted domain is shown. We work with newspapers' articles in Spanish, that is why every reference we use is in Spanish. For the identification and recognition of temporal expressions we base on a temporal expression grammar and for the resolution on a dictionary, where we have the information necessary to do the date operation based on the recognized expressions. In the evaluation of our proposal we have obtained successful results for the examples studied.

Riconoscimento delle emozioni tramite strumentazione per EEG

Lo scopo di questa trattazione è quindi di illustrare il lavoro svolto nel tentativo di classificare le reazioni emozionali ad immagini con una forte carica emozionale, sia positiva che negativa. A tale scopo sono stati acquisiti i segnali EEG di diversi soggetti durante l’esposizione ad immagini di vario contenuto, insieme alla loro reazione dichiarata alle immagini stesse. Queste sono state immagazzinate, elaborate utilizzando diversi metodi di estrazione delle informazioni, ed infine si è tentato di effettuare un riconoscimento di pattern sui segnali tramite algoritmi di apprendimento supervisionato; i dati sono stati quindi divisi tra dati di “training”, utilizzati per la strutturazione dell’algoritmo, e dati di test, necessari per la verifica dell’affidabilità dell’algoritmo.

Assessing facial approximation accuracy: How do resemblance ratings of disparate faces compare to recognition tests?

In the past, the accuracy of facial approximations has been assessed by resemblance ratings (i.e., the comparison of a facial approximation directly to a target individual) and recognition tests (e.g., the comparison of a facial approximation to a photo array of faces including foils and a target individual). Recently, several research studies have indicated that recognition tests hold major strengths in contrast to resemblance ratings. However, resemblance ratings remain popularly employed and/or are given weighting when judging facial approximations, thus indicating that no consensus has been reached. This study aims to further investigate the matter by comparing the results of resemblance ratings and recognition tests for two facial approximations which clearly differed in their morphological appearance. One facial approximation was constructed by an experienced practitioner privy to the appearance of the target individual (practitioner had direct access to an antemortem frontal photograph during face construction), while the other facial approximation was constructed by a novice under blind conditions. Both facial approximations, whilst clearly morphologically different, were given similar resemblance scores even though recognition test results produced vastly different results. One facial approximation was correctly recognized almost without exception while the other was not correctly recognized above chance rates. These results suggest that resemblance ratings are insensitive measures of the accuracy of facial approximations and lend further weight to the use of recognition tests in facial approximation assessment. (c) 2006 Elsevier Ireland Ltd. All rights reserved.

AÇÃO PASTORAL DA IGREJA CATÓLICA APOSTÓLICA ROMANA FACE AO DIREITO À INSERÇÃO SOCIAL DE PESSOAS EM SITUAÇÃO DE RUA

A proposta do presente estudo é verificar a ação pastoral da Igreja Católica junto ao povo de rua da cidade de São Paulo, tendo como objetivo formar um conceito teórico sobre a contribuição social da pastoral em um contexto urbano, a partir da ação de Entid ades de apoio ao povo de rua. A metodologia utilizada foi a bibliográfica. As implicações do estudo foram o direcionamento que a práxis pastoral está direcionada à priorização da superação e do reconhecimento da necessidade material e psicossocial de quem está morando na rua. A concretização da práxis se dá por meio de uma prática interventora sócio-politica, a qual visa a efetivação de medidas públicas para uma demanda de pessoas que usam a rua como moradia. A ação pastoral contribui ao mostrar a ausência de política pública que dificulta o reconhecimento deste grupo social como pessoas capacitadas a produzir e pertencer a sociedade em geral. E, ao mesmo tempo em que aponta a lacuna exposta pelo poder público, o agir pastoral sinaliza alternativa para o reconhecimento de pessoas que moram na rua com parcerias entre entidades não governamentais e movimentos sociais, como o MST, sendo assim uma via de reinserção social, além da promoção de Fóruns para a criação de medidas públicas com participação direta de pessoas que vivem na rua e albergues da cidade de São Paulo. Portanto, verifica-se uma práxis pastoral fundamentada por uma responsabilidade social dinamizada pela prática de parceria participativa que envolva as diversas esferas sociais para efetivação concreta dos direitos sociais da pessoa em situação de rua que vive em áreas urbana como a Cidade de São Paulo.

Character recognition systems using Fourier transformation in incoherent light

Many attempts have been made to overcome problems involved in character recognition which have resulted in the manufacture of character reading machines. An investigation into a new approach to character recognition is described. Features for recognition are Fourier coefficients. These are generated optically by convolving characters with periodic gratings. The development of hardware to enable automatic measurement of contrast and position of periodic shadows produced by the convolution is described. Fourier coefficients of character sets were measured, many of which are tabulated. Their analysis revealed that a few low frequency sampling points could be selected to recognise sets of numerals. Limited treatment is given to show the effect of type face variations on the values of coefficients which culminated in the location of six sampling frequencies used as features to recognise numerals in two type fonts. Finally, the construction of two character recognition machines is compared and contrasted. The first is a pilot plant based on a test bed optical Fourier analyser, while the second is a more streamlined machine d(3signed for high speed reading. Reasons to indicate that the latter machine would be the most suitable to adapt for industrial and commercial applications are discussed.

Contrast- and illumination-invariant object recognition from active sensation

It has been suggested that the deleterious effect of contrast reversal on visual recognition is unique to faces, not objects. Here we show from priming, supervised category learning, and generalization that there is no such thing as general invariance of recognition of non-face objects against contrast reversal and, likewise, changes in direction of illumination. However, when recognition varies with rendering conditions, invariance may be restored, and effects of continuous learning may be reduced, by providing prior object knowledge from active sensation. Our findings suggest that the degree of contrast invariance achieved reflects functional characteristics of object representations learned in a task-dependent fashion.

The heteromeric 5-HT3A/B receptor:the effect of 5-HT3B subunits on receptor structure and ligand recognition

The 5-HT3 receptors are members of the cys-loop family of ligand-gated ion channels. Two functional subtypes are known, the homomeric 5HT3A and the heteromeric 5HT3A/B receptors, which exhibit distinct biophysical characteristics but are difficult to differentiate pharmacologically. Atomic force microscopy has been used to determine the stoichiometry and architecture of the heteromeric 5HT3A/B receptor. Each subunit was engineered to express a unique C-terminal epitope tag, together with six sequential histidine residues to facilitate nickel affinity purification. The 5-HT3 receptors, ectopically expressed in HEK293 cells, were solubilised, purified and decorated with antibodies to the subunit specific epitope tags. Imaging of individual receptors by atomic force microscopy revealed a pentameric arrangement of subunits in the order BBABA, reading anti-clockwise when viewed from the extracellular face. Homology models for the heteromeric receptor were then constructed using both the electron microscopic structure of the nicotinic acetylcholine receptor, from Torpedo marmorata, and the X-ray crystallographic structure of the soluble acetylcholine binding protein, from Lymnaea stagnalis, as templates. These homology models were used, together with equivalent models constructed for the homomeric receptor, to interpret mutagenesis experiments designed to explore the minimal recognition differences of both the natural agonist, 5-HT, and the competitive antagonist, granisetron, for the two human receptor subtypes. The results of this work revealed that the 5-HT3B subunit residues within the ligand binding site, for both the agonist and antagonist, are accommodating to conservative mutations. They are consistent with the view that the 5-HT3A subunit provides the principal and the 5-HT38 subunit the complementary recognition interactions at the binding interface.

Footwear for gender recognition

Conventionally, biometrics resources, such as face, gait silhouette, footprint, and pressure, have been utilized in gender recognition systems. However, the acquisition and processing time of these biometrics data makes the analysis difficult. This letter demonstrates for the first time how effective the footwear appearance is for gender recognition as a biometrics resource. A footwear database is also established with reprehensive shoes (footwears). Preliminary experimental results suggest that footwear appearance is a promising resource for gender recognition. Moreover, it also has the potential to be used jointly with other developed biometrics resources to boost performance.

Dimensions of perception and recognition of danger

Despite abundant literature on human behaviour in the face of danger, much remains to be discovered. Some descriptive models of behaviour in the face of danger are reviewed in order to identify areas where documentation is lacking. It is argued that little is known about recognition and assessment of danger and yet, these are important aspects of cognitive processes. Speculative arguments about hazard assessment are reviewed and tested against the results of previous studies. Once hypotheses are formulated, the reason for retaining the reportory grid as the main research instrument are outlined, and the choice of data analysis techniques is described. Whilst all samples used repertory grids, the rating scales were different between samples; therefore, an analysis is performed of the way in which rating scales were used in the various samples and of some reasons why the scales were used differently. Then, individual grids are looked into and compared between respondents within each sample; consensus grids are also discussed. the major results from all samples are then contrasted and compared. It was hypothesized that hazard assessment would encompass three main dimensions, i.e. 'controllability', 'severity of consequences' and 'likelihood of occurrence', which would emerge in that order. the results suggest that these dimensions are but facets of two broader dimensions labelled 'scope of human intervention' and 'dangerousness'. It seems that these two dimensions encompass a number of more specific dimensions some of which can be further fragmented. Thus, hazard assessment appears to be a more complex process about which much remains to be discovered. Some of the ways in which further discovery might proceed are discussed.

Development of configural 3D object recognition

There is evidence for the late development in humans of configural face and animal recognition. We show that the recognition of artificial three-dimensional (3D) objects from part configurations develops similarly late. We also demonstrate that the cross-modal integration of object information reinforces the development of configural recognition more than the intra-modal integration does. Multimodal object representations in the brain may therefore play a role in configural object recognition. © 2003 Elsevier B.V. All rights reserved.

Late development of metric part-relational processing in object recognition

Four experiments with unfamiliar objects examined the remarkably late consolidation of part-relational relative to part-based object recognition (Jüttner, Wakui, Petters, Kaur, & Davidoff, 2013). Our results indicate a particularly protracted developmental trajectory for the processing of metric part relations. Schoolchildren aged 7 to 14 years and adults were tested in 3-Alternative-Forced-Choice tasks to judge the correct appearance of upright and inverted newly learned multipart objects that had been manipulated in terms of individual parts or part relations. Experiment 1 showed that even the youngest tested children were close to adult levels of performance for recognizing categorical changes of individual parts and relative part position. By contrast, Experiment 2 demonstrated that performance for detecting metric changes of relative part position was distinctly reduced in young children compared with recognizing metric changes of individual parts, and did not approach the latter until 11 to 12 years. A similar developmental dissociation was observed in Experiment 3, which contrasted the detection of metric relative-size changes and metric part changes. Experiment 4 showed that manipulations of metric size that were perceived as part (rather than part-relational) changes eliminated this dissociation. Implications for theories of object recognition and similarities to the development of face perception are discussed. © 2014 American Psychological Association.