Development of a hand-gesture recognition system for human-computer interaction

The aim of this Master Thesis is the analysis, design and development of a robust and reliable Human-Computer Interaction interface, based on visual hand-gesture recognition. The implementation of the required functions is oriented to the simulation of a classical hardware interaction device: the mouse, by recognizing a specific hand-gesture vocabulary in color video sequences. For this purpose, a prototype of a hand-gesture recognition system has been designed and implemented, which is composed of three stages: detection, tracking and recognition. This system is based on machine learning methods and pattern recognition techniques, which have been integrated together with other image processing approaches to get a high recognition accuracy and a low computational cost. Regarding pattern recongition techniques, several algorithms and strategies have been designed and implemented, which are applicable to color images and video sequences. The design of these algorithms has the purpose of extracting spatial and spatio-temporal features from static and dynamic hand gestures, in order to identify them in a robust and reliable way. Finally, a visual database containing the necessary vocabulary of gestures for interacting with the computer has been created.

"CRISE DE IDENTIDADE NO JORNALISMO" Pensamentos e posicionamentos de jornalistas em tempos difíceis

Esta pesquisa estuda o fenômeno crise de identidade do jornalista e do Jornalismo motivada pelas mudanças conjunturais oferecidas e impostas a esta área, em sua trajetória brasileira, de variadas formas e em diversos momentos. A evolução da sociedade e dos meios, as inovações técnicas e tecnológicas, a necessidade de formação, bem como os ataques à exigência do diploma e o cancelamento deste em nome do direito ao livre pensamento, entre outros. Por conta disso, busca-se a compreensão do indivíduo jornalista e da construção de sua identidade, bem como da sustentação de sua imagem, aspectos que ajudam a perceber sua importância na tessitura social. O esforço em localizar elementos da identidade e do status desse profissional considerou a análise de discursos de vários autores, no traçado que vai da década de 50 à contemporaneidade (ano de 2005). Para tanto recorreu-se às seguintes ferramentas: o aporte teórico relacionado à vontade de potência sugerida por Friedrich Nietzsche e a Ação Comunicativa de Jürgen Habermas. Do primeiro, adotou-se um modelo híbrido relacionando unidades-temáticas e sentidos de valor; do segundo procurou-se a interface do diálogo mantido por meio de parâmetros normativos, de vivência, e da memória armazenada pelos jornalistas no decorrer do tempo. Os resultados obtidos mostram, nas três fases adotadas, (décs. 50-60, 70-80, e 90-05) a recorrência aos valores ligados à liberdade e à ética, em variadas proporções e intensidades. A questão técnica foi praticamente deixada de lado, parecendo ter sido assimilada, ao passo que as relacionadas à formação foram acionadas esporadicamente. Também percebe-se em variados momentos e de variadas formas, os ataques ao jornalista, geralmente desqualificando-o ou colocando-o sob suspeita. Por outro lado, em todos, os casos os jornalistas procuram defender-se sempre mobilizando recursos, que na maioria das vezes deixam abertas as possibilidades ao diálogo, assim como tentam recuperar sua importância mostrando seus valores por meio da capacidade que possuem de representar a sociedade.(AU)

"CRISE DE IDENTIDADE NO JORNALISMO" Pensamentos e posicionamentos de jornalistas em tempos difíceis

Esta pesquisa estuda o fenômeno crise de identidade do jornalista e do Jornalismo motivada pelas mudanças conjunturais oferecidas e impostas a esta área, em sua trajetória brasileira, de variadas formas e em diversos momentos. A evolução da sociedade e dos meios, as inovações técnicas e tecnológicas, a necessidade de formação, bem como os ataques à exigência do diploma e o cancelamento deste em nome do direito ao livre pensamento, entre outros. Por conta disso, busca-se a compreensão do indivíduo jornalista e da construção de sua identidade, bem como da sustentação de sua imagem, aspectos que ajudam a perceber sua importância na tessitura social. O esforço em localizar elementos da identidade e do status desse profissional considerou a análise de discursos de vários autores, no traçado que vai da década de 50 à contemporaneidade (ano de 2005). Para tanto recorreu-se às seguintes ferramentas: o aporte teórico relacionado à vontade de potência sugerida por Friedrich Nietzsche e a Ação Comunicativa de Jürgen Habermas. Do primeiro, adotou-se um modelo híbrido relacionando unidades-temáticas e sentidos de valor; do segundo procurou-se a interface do diálogo mantido por meio de parâmetros normativos, de vivência, e da memória armazenada pelos jornalistas no decorrer do tempo. Os resultados obtidos mostram, nas três fases adotadas, (décs. 50-60, 70-80, e 90-05) a recorrência aos valores ligados à liberdade e à ética, em variadas proporções e intensidades. A questão técnica foi praticamente deixada de lado, parecendo ter sido assimilada, ao passo que as relacionadas à formação foram acionadas esporadicamente. Também percebe-se em variados momentos e de variadas formas, os ataques ao jornalista, geralmente desqualificando-o ou colocando-o sob suspeita. Por outro lado, em todos, os casos os jornalistas procuram defender-se sempre mobilizando recursos, que na maioria das vezes deixam abertas as possibilidades ao diálogo, assim como tentam recuperar sua importância mostrando seus valores por meio da capacidade que possuem de representar a sociedade.(AU)

Membrane-bound state of the colicin E1 channel domain as an extended two-dimensional helical array

Atomic level structures have been determined for the soluble forms of several colicins and toxins, but the structural changes that occur after membrane binding have not been well characterized. Changes occurring in the transition from the soluble to membrane-bound state of the C-terminal 190-residue channel polypeptide of colicin E1 (P190) bound to anionic membranes are described. In the membrane-bound state, the α-helical content increases from 60–64% to 80–90%, with a concomitant increase in the average length of the helical segments from 12 to 16 or 17 residues, close to the length required to span the membrane bilayer in the open channel state. The average distance between helical segments is increased and interhelix interactions are weakened, as shown by a major loss of tertiary structure interactions, decreased efficiency of fluorescence resonance energy transfer from an energy donor on helix V of P190 to an acceptor on helix IX, and decreased resonance energy transfer at higher temperatures, not observed in soluble P190, implying freedom of motion of helical segments. Weaker interactions are also shown by a calorimetric thermal transition of low cooperativity, and the extended nature of the helical array is shown by a 3- to 4-fold increase in the average area subtended per molecule to 4,200 Å2 on the membrane surface. The latter, with analysis of the heat capacity changes, implies the absence of a developed hydrophobic core in the membrane-bound P190. The membrane interfacial layer thus serves to promote formation of a highly helical extended two-dimensional flexible net. The properties of the membrane-bound state of the colicin channel domain (i.e., hydrophobic anchor, lengthened and loosely coupled α-helices, and close association with the membrane interfacial layer) are plausible structural features for the state that is a prerequisite for voltage gating, formation of transmembrane helices, and channel opening.

The EF-hand Ca2+-binding Protein p22 Associates with Microtubules in an N-Myristoylation–dependent Manner

Proteins containing the EF-hand Ca2+-binding motif, such as calmodulin and calcineurin B, function as regulators of various cellular processes. Here we focus on p22, an N-myristoylated, widely expressed EF-hand Ca2+-binding protein conserved throughout evolution, which was shown previously to be required for membrane traffic. Immunofluorescence studies show that p22 distributes along microtubules during interphase and mitosis in various cell lines. Moreover, we report that p22 associates with the microtubule cytoskeleton indirectly via a cytosolic microtubule-binding factor. Gel filtration studies indicate that the p22–microtubule-binding activity behaves as a 70- to 30-kDa globular protein. Our results indicate that p22 associates with microtubules via a novel N-myristoylation–dependent mechanism that does not involve classic microtubule-associated proteins and motor proteins. The association of p22 with microtubules requires the N-myristoylation of p22 but does not involve p22’s Ca2+-binding activity, suggesting that the p22–microtubule association and the role of p22 in membrane traffic are functionally related, because N-myristoylation is required for both events. Therefore, p22 is an excellent candidate for a protein that can mediate interactions between the microtubule cytoskeleton and membrane traffic.

Stretching single-domain proteins: Phase diagram and kinetics of force-induced unfolding

Single-molecule force spectroscopy reveals unfolding of domains in titin on stretching. We provide a theoretical framework for these experiments by computing the phase diagrams for force-induced unfolding of single-domain proteins using lattice models. The results show that two-state folders (at zero force) unravel cooperatively, whereas stretching of non-two-state folders occurs through intermediates. The stretching rates of individual molecules show great variations reflecting the heterogeneity of force-induced unfolding pathways. The approach to the stretched state occurs in a stepwise “quantized” manner. Unfolding dynamics and forces required to stretch proteins depend sensitively on topology. The unfolding rates increase exponentially with force f till an optimum value, which is determined by the barrier to unfolding when f = 0. A mapping of these results to proteins shows qualitative agreement with force-induced unfolding of Ig-like domains in titin. We show that single-molecule force spectroscopy can be used to map the folding free energy landscape of proteins in the absence of denaturants.

A genomic approach to gene fusion technology

Gene expression profiling provides powerful analyses of transcriptional responses to cellular perturbation. In contrast to DNA array-based methods, reporter gene technology has been underused for this application. Here we describe a genomewide, genome-registered collection of Escherichia coli bioluminescent reporter gene fusions. DNA sequences from plasmid-borne, random fusions of E. coli chromosomal DNA to a Photorhabdus luminescens luxCDABE reporter allowed precise mapping of each fusion. The utility of this collection covering about 30% of the transcriptional units was tested by analyzing individual fusions representative of heat shock, SOS, OxyR, SoxRS, and cya/crp stress-responsive regulons. Each fusion strain responded as anticipated to environmental conditions known to activate the corresponding regulatory circuit. Thus, the collection mirrors E. coli's transcriptional wiring diagram. This genomewide collection of gene fusions provides an independent test of results from other gene expression analyses. Accordingly, a DNA microarray-based analysis of mitomycin C-treated E. coli indicated elevated expression of expected and unanticipated genes. Selected luxCDABE fusions corresponding to these up-regulated genes were used to confirm or contradict the DNA microarray results. The power of partnering gene fusion and DNA microarray technology to discover promoters and define operons was demonstrated when data from both suggested that a cluster of 20 genes encoding production of type I extracellular polysaccharide in E. coli form a single operon.

High-sensitivity array analysis of gene expression for the early detection of disseminated breast tumor cells in peripheral blood

Early detection is an effective means of reducing cancer mortality. Here, we describe a highly sensitive high-throughput screen that can identify panels of markers for the early detection of solid tumor cells disseminated in peripheral blood. The method is a two-step combination of differential display and high-sensitivity cDNA arrays. In a primary screen, differential display identified 170 candidate marker genes differentially expressed between breast tumor cells and normal breast epithelial cells. In a secondary screen, high-sensitivity arrays assessed expression levels of these genes in 48 blood samples, 22 from healthy volunteers and 26 from breast cancer patients. Cluster analysis identified a group of 12 genes that were elevated in the blood of cancer patients. Permutation analysis of individual genes defined five core genes (P ≤ 0.05, permax test). As a group, the 12 genes generally distinguished accurately between healthy volunteers and patients with breast cancer. Mean expression levels of the 12 genes were elevated in 77% (10 of 13) untreated invasive cancer patients, whereas cluster analysis correctly classified volunteers and patients (P = 0.0022, Fisher's exact test). Quantitative real-time PCR confirmed array results and indicated that the sensitivity of the assay (1:2 × 108 transcripts) was sufficient to detect disseminated solid tumor cells in blood. Expression-based blood assays developed with the screening approach described here have the potential to detect and classify solid tumor cells originating from virtually any primary site in the body.

An orchestrated gene expression component of neuronal programmed cell death revealed by cDNA array analysis

Programmed cell death (PCD) during neuronal development and disease has been shown to require de novo RNA synthesis. However, the time course and regulation of target genes is poorly understood. By using a brain-biased array of over 7,500 cDNAs, we profiled this gene expression component of PCD in cerebellar granule neurons challenged separately by potassium withdrawal, combined potassium and serum withdrawal, and kainic acid administration. We found that hundreds of genes were significantly regulated in discreet waves including known genes whose protein products are involved in PCD. A restricted set of genes was regulated by all models, providing evidence that signals inducing PCD can regulate large assemblages of genes (of which a restricted subset may be shared in multiple pathways).

Induction and repression of DAN1 and the family of anaerobic mannoprotein genes in Saccharomyces cerevisiae occurs through a complex array of regulatory sites

The DAN/TIR mannoprotein genes of Saccharomyces cerevisiae (DAN1, DAN2, DAN3, DAN4, TIR1, TIR2, TIR3 and TIR4) are expressed in anaerobic cells while the predominant cell wall proteins Cwp1 and Cwp2 are down-regulated. Elements involved in activation and repression of the DAN/TIR genes were defined in this study, using the DAN1 promoter as a model. Nested deletions in a DAN1/lacZ reporter pinpointed regions carrying activation and repression elements. Inspection revealed two consensus sequences subsequently shown to be independent anaerobic response elements (AR1, consensus TCGTTYAG; AR2, consensus AAAAATTGTTGA). AR1 is found in all of the DAN/TIR promoters; AR2 is found in DAN1, DAN2 and DAN3. A 120 bp segment carrying two copies of AR1 preferentially activated transcription of lacZ under anaerobic conditions. A fusion of three synthetic copies of AR1 to MEL1 was also expressed anaerobically. Mutations in either AR1 site within the 120 bp segment caused a drastic loss of expression, indicating that both are necessary for activation and implying cooperativity between adjacent transcriptional activation complexes. A single AR2 site carried on a 46 bp fragment from the DAN1 promoter activated lacZ transcription under anaerobic conditions, as did a 26 bp synthetic AR2 fragment fused to MEL1. Nucleotide substitutions within the AR2 sequence eliminated the activity of the 46 bp segment. Ablation of the AR2 sequences in the full promoter caused a partial reduction of expression. The presence of the ATTGTT core (recognized by HMG proteins) in the AR2 sequence suggests that an HMG protein may activate through AR2. One region was implicated in aerobic repression of DAN1. It contains sites for the heme-induced Mot3 and Rox1 repressors.

Behavioral inferences from the Skhul/Qafzeh early modern human hand remains

Two groups of humans are found in the Near East ≈100,000 years ago, the late archaic Neanderthals and the early modern Skhul/Qafzeh humans. Observations that Neanderthals were more heavily muscled, had stronger upper-limb bones, and possessed unusual shapes and orientations of some upper-limb joint complexes relative to the Skhul/Qafzeh hominids, have led some researchers to conclude that significant between-group upper-limb-related behavioral differences must have been present, despite the association of the two groups with similar Middle Paleolithic archeological complexes. A three-dimensional morphometric analysis of the hand remains of the Skhul/Qafzeh hominids, Neanderthals, early and late Upper Paleolithic humans, and Holocene humans supports the dichotomy. The Skhul/Qafzeh carpometacarpal remains do not have any unique morphologies relative to the other fossil samples remains examined. However, in the functionally significant metacarpal 1 and 3 bases they resemble Upper Paleolithic humans, not Neanderthals. Furthermore, the Skhul/Qafzeh sample differs significantly from the Neanderthals in many other aspects of hand functional anatomy. Given the correlations between changes in tool technologies and functional adaptations seen in the hands of Upper Paleolithic humans, it is concluded that the Skhul/Qafzeh hand remains were adapted to Upper Paleolithic-like manipulative repertoires. These results support the inference of significant behavioral differences between Neanderthals and the Skhul/Qafzeh hominids and indicate that a significant shift in human manipulative behaviors was associated with the earliest stages of the emergence of modern humans.