Neuronal mechanism of state control in Drosophila melanogaster

The changes in internal states, such as fear, hunger and sleep affect behavioral responses in animals. In most of the cases, these state-dependent influences are “pleiotropic”: one state affects multiple sensory modalities and behaviors; “scalable”: the strengths and choices of such modulations differ depending on the imminence of demands; and “persistent”: once the state is switched on the effects last even after the internal demands are off. These prominent features of state-control enable animals to adjust their behavioral responses depending on their internal demands. Here, we studied the neuronal mechanisms of state-controls by investigating energy-deprived state (hunger state) and social-deprived state of fruit flies, Drosophila melanogaster, as prototypic models. To approach these questions, we developed two novel methods: a genetically based method to map sites of neuromodulation in the brain and optogenetic tools in Drosophila.

These methods, and genetic perturbations, reveal that the effect of hunger to alter behavioral sensitivity to gustatory cues is mediate by two distinct neuromodulatory pathways. The neuropeptide F (NPF) – dopamine (DA) pathway increases sugar sensitivity under mild starvation, while the adipokinetic hormone (AKH)- short neuropeptide F (sNPF) pathway decreases bitter sensitivity under severe starvation. These two pathways are recruited under different levels of energy demands without any cross interaction. Effects of both of the pathways are mediated by modulation of the gustatory sensory neurons, which reinforce the concept that sensory neurons constitute an important locus for state-dependent control of behaviors. Our data suggests that multiple independent neuromodulatory pathways are underlying pleiotropic and scalable effects of the hunger state.

In addition, using optogenetic tool, we show that the neural control of male courtship song can be separated into probabilistic/biasing, and deterministic/command-like components. The former, but not the latter, neurons are subject to functional modulation by social experience, supporting the idea that they constitute a locus of state-dependent influence. Interestingly, moreover, brief activation of the former, but not the latter, neurons trigger persistent behavioral response for more than 10 min. Altogether, these findings and new tools described in this dissertation offer new entry points for future researchers to understand the neuronal mechanism of state control.

Z-selective olefin metathesis using chelating ruthenium alkylidene catalysts

Publications about olefin metathesis will generally discuss how the discovery and development of well-defined catalysts to carry out this unique transformation have revolutionized many fields, from natural product and materials chemistry, to green chemistry and biology. However, until recently, an entire manifestation of this methodology had been inaccessible. Except for a few select examples, metathesis catalysts favor the thermodynamic trans- or E-olefin products in cross metathesis (CM), macrocyclic ring closing metathesis (mRCM), ring opening metathesis polymerization (ROMP), and many other types of reactions. Judicious choice of substrates had allowed for the direct synthesis of cis- or Z-olefins or species that could be converted upon further reaction, however the catalyst controlled synthesis of Z-olefins was not possible until very recently.

Research into the structure and stability of metallacyclobutane intermediates has led to the proposal of models to impart Z-selectivity in metathesis reactions. Having the ability to influence the orientation of metallacyclobutane substituents to cause productive formation of Z- double bonds using steric and electronic effects was highly desired. The first successful realization of this concept was by Schrock and Hoveyda et al. who synthesized monoaryloxide pyrolidine (MAP) complexes of tungsten and molybdenum that promoted Z-selective CM. The Z-selectivity of these catalysts was attributed to the difference in the size of the two axial ligands. This size difference influences the orientation of the substituents on the forming/incipient metallacyclobutane intermediate to a cis-geometry and leads to productive formation of Z-olefins. These catalysts have shown great utility in the synthesis of complicated natural product precursors and stereoregular polymers. More recently, ruthenium catalysts capable of promoting Z-selective metathesis have been reported by our group and others. This thesis will discuss the development of ruthenium-based NHC chelated Z-selective catalysts, studies probing their unique metathesis mechanism, and synthetic applications that have been investigated thus far.

Chapter 1 will focus on studies into the stability of NHC chelated complexes and the synthesis of new and improved stable chelating architectures. Chapter 2 will discuss applications of the highly active and Z-selective developed in Chapter 1, including the formation of lepidopteran female sex pheromones using olefin cross metathesis and highly Z- and highly E-macrocycles using macrocyclic ring closing metathesis and Z-selective ethenolysis. Chapter 3 will explore studies into the unique mechanism of olefin metathesis reactions catalyzed by these NHC chelated, highly Z-selective catalysts, explaining observed trends by investigating the stability of relevant, substituted metallacyclobutane intermediates.

Brf1 post-transcriptionally regulates pluripotency and differentiation responses downstream of Erk MAP Kinase

FGF/Erk MAP Kinase Signaling is a central regulator of mouse embryonic stem cell (mESC) self-renewal, pluripotency and differentiation. However, the mechanistic connection between this signaling pathway activity and the gene circuits stabilizing mESCs in vitro remain unclear. Here we show that FGF signaling post-transcriptionally regulates the mESC transcription factor network by controlling the expression of Brf1 (zfp36l1), an AU-rich element mRNA binding protein. Changes in Brf1 level disrupts the expression of core pluripotency-associated genes and attenuates mESC self-renewal without inducing differentiation. These regulatory effects are mediated by rapid and direct destabilization of Brf1 targets, such as Nanog mRNA. Interestingly, enhancing Brf1 expression does not compromise mESC pluripotency, but does preferentially regulate differentiation to mesendoderm by accelerating the expression of primitive streak markers. Together, these studies demonstrate that FGF signals utilize targeted mRNA degradation by Brf1 to enable rapid post-transcriptional control of gene expression.

Sensitivity map of laser tweezers Raman spectroscopy for single-cell analysis of colorectal cancer

Raman spectroscopy on single, living epithelial cells captured in a laser trap is shown to have diagnostic power over colorectal cancer. This new single-cell technology comprises three major components: primary culture processing of human tissue samples to produce single-cell suspensions, Raman detection on singly trapped cells, and diagnoses of the cells by artificial neural network classifications. it is compared with DNA flow cytometry for similarities and differences. Its advantages over tissue Raman spectroscopy are also discussed. In the actual construction of a diagnostic model for colorectal cancer, real patient data were taken to generate a training set of 320 Raman spectra and, a test set of 80. By incorporating outlier corrections to a conventional binary neural classifier, our network accomplished significantly better predictions than logistic regressions, with sensitivity improved from 77.5% to 86.3% and specificity improved from 81.3% to 86.3% for the training set and moderate improvements for the test set. Most important, the network approach enables a sensitivity map analysis to quantitate the relevance of each Raman band to the normal-to-cancer transform at the cell level. Our technique has direct clinic applications for diagnosing cancers and basic science potential in the study of cell dynamics of carcinogenesis. (C) 2007 Society of Photo-Optical Instrumentation Engineers.

The Semiglades: the collision of restoration, social values, and the ecosystem concept

Opinion article

Sensitivity map of laser tweezers Raman spectroscopy for single-cell analysis of colorectal cancer

Raman spectroscopy on single, living epithelial cells captured in a laser trap is shown to have diagnostic power over colorectal cancer. This new single-cell technology comprises three major components: primary culture processing of human tissue samples to produce single-cell suspensions, Raman detection on singly trapped cells, and diagnoses of the cells by artificial neural network classifications. it is compared with DNA flow cytometry for similarities and differences. Its advantages over tissue Raman spectroscopy are also discussed. In the actual construction of a diagnostic model for colorectal cancer, real patient data were taken to generate a training set of 320 Raman spectra and, a test set of 80. By incorporating outlier corrections to a conventional binary neural classifier, our network accomplished significantly better predictions than logistic regressions, with sensitivity improved from 77.5% to 86.3% and specificity improved from 81.3% to 86.3% for the training set and moderate improvements for the test set. Most important, the network approach enables a sensitivity map analysis to quantitate the relevance of each Raman band to the normal-to-cancer transform at the cell level. Our technique has direct clinic applications for diagnosing cancers and basic science potential in the study of cell dynamics of carcinogenesis. (C) 2007 Society of Photo-Optical Instrumentation Engineers.

Novel method to construct a quality map for phase unwrapping based on modulation and the phase gradient

A novel method to construct a quality map, called modulation-phase-gradient variance (MPGV), is proposed, based on modulation and the phase gradient. The MPGV map is successfully applied to two phase-unwrapping algorithms - the improved weighted least square and the quality-guided unwrapping algorithm. Both simulated and experimental data testify to the validity of our proposed quality map. Moreover, the unwrapped-phase results show that the new quality map can have higher reliability than the conventional phase-derivative variance quality map in helping to unwrap noisy, low-modulation, and/or discontinuous phase maps. (c) 2006 Society of Photo-Optical Instrumentation Engineers.

A reliable phase unwrapping algorithm based on the local fitting plane and quality map

A fast and reliable phase unwrapping (PhU) algorithm, based on the local quality-guided fitting plane, is presented. Its framework depends on the basic plane-approximated assumption for phase values of local pixels and on the phase derivative variance (PDV) quality map. Compared with other existing popular unwrapping algorithms, the proposed algorithm demonstrated improved robustness and immunity to strong noise and high phase variations, given that the plane assumption for local phase is reasonably satisfied. Its effectiveness is demonstrated by computer-simulated and experimental results.

Novel method to construct a quality map for phase unwrapping based on modulation and the phase gradient

A novel method to construct a quality map, called modulation-phase-gradient variance (MPGV), is proposed, based on modulation and the phase gradient. The MPGV map is successfully applied to two phase-unwrapping algorithms - the improved weighted least square and the quality-guided unwrapping algorithm. Both simulated and experimental data testify to the validity of our proposed quality map. Moreover, the unwrapped-phase results show that the new quality map can have higher reliability than the conventional phase-derivative variance quality map in helping to unwrap noisy, low-modulation, and/or discontinuous phase maps. (c) 2006 Society of Photo-Optical Instrumentation Engineers.

Bizimodu osasungarrien sustapenerako marketin plana: Athlon E. Koop David Spine Concept

marketin plana

Weighted least-squares phase unwrapping algorithm based on derivative variance correlation map

Among different phase unwrapping approaches, the weighted least-squares minimization methods are gaining attention. In these algorithms, weighting coefficient is generated from a quality map. The intrinsic drawbacks of existing quality maps constrain the application of these algorithms. They often fail to handle wrapped phase data contains error sources, such as phase discontinuities, noise and undersampling. In order to deal with those intractable wrapped phase data, a new weighted least-squares phase unwrapping algorithm based on derivative variance correlation map is proposed. In the algorithm, derivative variance correlation map, a novel quality map, can truly reflect wrapped phase quality, ensuring a more reliable unwrapped result. The definition of the derivative variance correlation map and the principle of the proposed algorithm are present in detail. The performance of the new algorithm has been tested by use of a simulated spherical surface wrapped data and an experimental interferometric synthetic aperture radar (IFSAR) wrapped data. Computer simulation and experimental results have verified that the proposed algorithm can work effectively even when a wrapped phase map contains intractable error sources. (c) 2006 Elsevier GmbH. All rights reserved.

As metáforas conceptuais da palavra paz nos relatórios do Conselho de Segurança das Nações Unidas: uma análise baseada em corpus digital

A presente pesquisa tem por objetivo investigar como a palavra paz é entendida, em termos de conceito, pelo Conselho de Segurança da Organização das Nações Unidas. Para tanto, são analisados trinta e sete relatórios oficiais produzidos pelo Conselho de Segurança, no período de agosto de 1994 a junho de 2009, acerca das missões de paz realizadas em trinta e uma regiões/países que apresentavam ameaça à paz e à segurança internacionais durante aquele período. De acordo com a Conselheira Gilda Santos Neves, chefe da Divisão das Nações Unidas do Ministério das Relações Exteriores, em seu texto O Brasil e a Criação da Comissão para a Consolidação da Paz (2008), a paz é algo que se consolida e não se constrói. Tal posição norteia a presente pesquisa, uma vez que o objetivo aqui é mapear as expressões linguísticas realizadas através da palavra paz. As bases teóricas desta pesquisa encontram-se fundamentadas na teoria da metáfora cognitiva, de Lakoff e Johnson (1980), bem como no estudo de Deignan (2005) em seu livro intitulado Metaphor and Corpus Linguistics, que visa a fornecer os benefícios que a abordagem cognitiva de metáforas pode obter através da análise de corpora digitalizados. Após compilar os relatórios do Conselho de Segurança e prepará-los para serem lidos pelo programa computacional WordSmith Tools 3.0, foram extraídas todas as ocorrências da palavra paz dos referidos relatórios. Das 686 ocorrências geradas, foram deixadas para análise somente aquelas com sentido metafórico e, no total, nove esquemas conceptuais foram construídos. A pesquisa feita sugere que, para o Conselho de Segurança, a paz é algo profundamente desejado tanto pela população das zonas de conflito quanto pela comunidade internacional. No entanto, a paz não é facilmente construída ou estabelecida. Alcançar a paz implica seguir um processo com diferentes etapas, ou seja, com início, meio e fim, bem como superar obstáculos e retrocessos que surgem no meio do caminho. Para tanto, diversos investimentos têm de ser feitos por todos aqueles envolvidos e realmente interessados na paz mundial. Por fim, vê-se que a visão da Conselheira Gilda Santos Neves, de acordo com as metáforas aqui analisadas, está correta, já que, conforme apontam os resultados do presente estudo, o conceito de paz, para o Conselho de Segurança, não é o de algo a ser construído do zero