Binding Site Structure and Stoichiometry in Serotonin Type 3 Receptors

This dissertation primarily describes studies of serotonin type 3 (5-HT₃) receptors of the Cys-loop super-family of ligand gated ion channels. The first chapter provides a general introduction to these important proteins and the methods used to interrogate their structure and function. The second chapter details the delineation of a structural unit of the ligand binding site of homomeric 5-HT₃A receptors on which the ligands serotonin (5-HT) and m-chlorophenyl biguanide (mCPBG) are reliant for effective receptor activation. Unnatural amino acid mutagenesis results show that the details of each ligand’s interaction with this organizing feature of the binding site differ, providing insights into general principles of receptor activation.

The third chapter describes a study in which florescent protein fusions of the A and B subunits of the heteromeric 5-HT₃AB receptor are employed to determine the subunit stoichiometry and order within functional receptors. Strong evidence is found for an A₃B₂ stoichiometry with A-A-B-A-B order. The fourth chapter investigates the potential for ligand binding across heteromeric binding sites in the 5-HT₃AB receptor. Unlike serotonin, mCPBG is found to bind the receptor at heteromeric binding sites. Further mCPBG is capable of allosterically modulating the response of serotonin on the 5-HT₃AB receptor from these heteromeric sites.

Finally, the fifth chapter describes progress towards the application of unnatural amino acid mutagenesis to an important new class of proteins, transcription factors. Experiments optimizing novel methods for the detection of function are described, using RARα of the nuclear receptor family of transcription factors.

Site-Specific Isotopes in Small Organic Molecules

Stable isotope geochemistry is a valuable toolkit for addressing a broad range of problems in the geosciences. Recent technical advances provide information that was previously unattainable or provide unprecedented precision and accuracy. Two such techniques are site-specific stable isotope mass spectrometry and clumped isotope thermometry. In this thesis, I use site-specific isotope and clumped isotope data to explore natural gas development and carbonate reaction kinetics. In the first chapter, I develop an equilibrium thermodynamics model to calculate equilibrium constants for isotope exchange reactions in small organic molecules. This equilibrium data provides a framework for interpreting the more complex data in the later chapters. In the second chapter, I demonstrate a method for measuring site-specific carbon isotopes in propane using high-resolution gas source mass spectrometry. This method relies on the characteristic fragments created during electron ionization, in which I measure the relative isotopic enrichment of separate parts of the molecule. My technique will be applied to a range of organic compounds in the future. For the third chapter, I use this technique to explore diffusion, mixing, and other natural processes in natural gas basins. As time progresses and the mixture matures, different components like kerogen and oil contribute to the propane in a natural gas sample. Each component imparts a distinct fingerprint on the site-specific isotope distribution within propane that I can observe to understand the source composition and maturation of the basin. Finally, in Chapter Four, I study the reaction kinetics of clumped isotopes in aragonite. Despite its frequent use as a clumped isotope thermometer, the aragonite blocking temperature is not known. Using laboratory heating experiments, I determine that the aragonite clumped isotope thermometer has a blocking temperature of 50-100°C. I compare this result to natural samples from the San Juan Islands that exhibit a maximum clumped isotope temperature that matches this blocking temperature. This thesis presents a framework for measuring site-specific carbon isotopes in organic molecules and new constraints on aragonite reaction kinetics. This study represents the foundation of a future generation of geochemical tools for the study of complex geologic systems.

Report on the Linda Vista bridge site

The Linda Vista Bridge spans the Arroyo Seco about a quarter of a mile above the Colorado Street Bridge, but serves an entirely different territory; as there is no road between there on the west bank. Los Angeles, Hollywood, and several of the beach cities can be reached by the way of the Colorado Street Bridge. The Linda Vista Bridge carries the traffic to the northwest of Pasadena, that is, Flintridge, Linda Vista, Montrose, Sunland. After leaving the bridge, the road follows the west bank of the Arroyo almost to the mouth of the canyon; then to the west along the foot of the mountains and into the San Fernando Valley.

Pollution and water quality control of the River Tejo. Report on a visit to Portugal, Sep 27th - October 10th, 1987

Fundamental changes in the management of water resources in Portugal are now evolving. Five regional organisations termed Administracaos de Regiao Hidrographic (ARH), will be created to manage water resources within their respective geographical areas. These areas will be catchment based. As a fore-runner to the implementation of the five ARH's a foundation project has been established within the Direcao-Geral do Recursos Naturais to examine the practical implications of the new system. This project has been divided into a number of sub-projects and complementary projects to include the Tejo complementary project. The Tejo complementary project is the focus of this report. The report is to advise on the role of biology in the proposed ARH, to establish priorities for biological studies within the present Projecto de Gestao Integrada dos Recursos Hidricos da Bacia Hidrografica do Rio Tejo (PGIRH/T) and to assist with the planning of laboratory facilities for biology at the new PGIRH/T laboratory at Alges, Lisboa.

I. Site effects and exciton structure in molecular crystals - Benzene. II. The first and second triplet states of solid benzene

The effect of intermolecular coupling in molecular energy levels (electronic and vibrational) has been investigated in neat and isotopic mixed crystals of benzene. In the isotopic mixed crystals of C₆H₆, C₆H₅D, m-C₆H₄D₂, p-C₆H₄D₂, sym-C₆H₃D₃, C₆D₅H, and C₆D₆ in either a C₆H₆ or C₆D₆ host, the following phenomena have been observed and interpreted in terms of a refined Frenkel exciton theory: a) Site shifts; b) site group splittings of the degenerate ground state vibrations of C₆H₆, C₆D₆, and sym-C₆H₃D₃; c) the orientational effect for the isotopes without a trigonal axis in both the ¹B_2u electronic state and the ground state vibrations; d) intrasite Fermi resonance between molecular fundamentals due to the reduced symmetry of the crystal site; and e) intermolecular or intersite Fermi resonance between nearly degenerate states of the host and guest molecules. In the neat crystal experiments on the ground state vibrations it was possible to observe many of these phenomena in conjunction with and in addition to the exciton structure.

To theoretically interpret these diverse experimental data, the concepts of interchange symmetry, the ideal mixed crystal, and site wave functions have been developed and are presented in detail. In the interpretation of the exciton data the relative signs of the intermolecular coupling constants have been emphasized, and in the limit of the ideal mixed crystal a technique is discussed for locating the exciton band center or unobserved exciton components. A differentiation between static and dynamic interactions is made in the Frenkel limit which enables the concepts of site effects and exciton coupling to be sharpened. It is thus possible to treat the crystal induced effects in such a fashion as to make their similarities and differences quite apparent.

A calculation of the ground state vibrational phenomena (site shifts and splittings, orientational effects, and exciton structure) and of the crystal lattice modes has been carried out for these systems. This calculation serves as a test of the approximations of first order Frenkel theory and the atom-atom, pair wise interaction model for the intermolecular potentials. The general form of the potential employed was V(r) = Be^-Cr - A/r⁶ ; the force constants were obtained from the potential by assuming the atoms were undergoing simple harmonic motion.

In part II the location and identification of the benzene first and second triplet states (³B_1u and ³E_1u) is given.

As práticas desenvolvidas no cuidado realizado por enfermeiros: um estudo de caso na rede básica de saúde no município do Rio de Janeiro

Esta pesquisa tem como objetivo geral, compreender as práticas desenvolvidas no cuidado realizado por enfermeiros nos Programas Nacionais de Saúde em uma unidade da rede básica de saúde no município do Rio de Janeiro. Este estudo corresponde ao desdobramento do projeto Práticas de cuidado no SUS: o papel do enfermeiro na Atenção Básica, apoiado por bolsa de produtividade em pesquisa da Universidade do Estado do Rio de Janeiro (UERJ/ Prociência). A presente dissertação trata-se de um estudo de caso realizado no âmbito da Atenção Básica, e, possui como cenário o Centro Municipal de Saúde (CMS) Milton Fontes Magarão. Os sujeitos da pesquisa corresponderam a oito enfermeiros, sendo sete entrevistas e um depoimento. Os instrumentos de coleta de dados utilizados, durante o período de julho a setembro de 2011, foram: observação sistemática, entrevista semi-estruturada, depoimento, formulário e pesquisa em fontes documentais. Para a avaliação dos resultados utilizou-se a técnica de análise temática, proposta por Minayo. No plano metodológico a análise foi qualitativa, no entanto, no tratamento dos dados, a análise aplicada se apropriou de elementos da abordagem quantitativa, com o objetivo de encontrar os núcleos de sentido contextualizados nas entrevistas. A partir dos resultados alcançados foi possível elaborar três categorias. A primeira categoria focou uma abordagem temática sobre as práticas desenvolvidas no cuidado realizado por enfermeiros nos Programas Nacionais. A segunda categoria buscou descrever a visão do enfermeiro sobre o desenvolvimento de suas práticas e do cuidado realizado, no qual, foram discutidas duas temáticas centrais: noções acerca das atividades realizadas pelos enfermeiros e noções dos enfermeiros acerca dos princípios e diretrizes para atenção à saúde. A terceira categoria direcionou-se para os fatores que interferem nas práticas e no cuidado realizado pelos enfermeiros. A prática e o cuidado realizados pelos enfermeiros foram analisados e discutidos, a partir dos relatos dos profissionais sobre suas vivências. Sendo assim, não coube a expressão de uma única forma de prática ou de uma única dimensão do cuidado, visto que essas práticas parecem estar entrelaçadas a diversos aspectos subjetivos, epistemológicos, culturais e sócio-econômicos. As práticas realizadas, independente do Programa Nacional de atuação dos enfermeiros, estão voltadas principalmente para a consulta de enfermagem, a prática educativa, a organização do fluxo de entrada, o sistema de referência e a visita domiciliar. Pôde-se inferir que diversos são os fatores que influenciam, de forma positiva ou negativa, o desenvolvimento das atividades diárias. Por fim, cabe ressaltar a necessidade de novos estudos sobre a temática a fim de estimular a construção de saberes teórico-práticos e a formulação de uma visão crítica sobre o cuidado profissional realizado pelo enfermeiro no âmbito da Atenção Básica.

Variations saisonnières des facteurs hydrologiques et du biovolume microalgal benthique au sein d'un habitat artificiel (acadja-enclos) dans la lagune Ebrié: site d'Adjapoté

This paper presents the results of a comparative study of chemical and physical characteristics (temperature, oxygen, salinity, turbidity, pH, nutrient: phosphates, nitrates, nitrites, ammoniac) within the extensive aquaculture structure (acadja-enclosure or artificial habitats for fishes) and in the surrounding water. Biological characteristics of the periphyton growing on bamboos from acadja-enclosure were also described.

Variations temporelles de Vibrio cholerae Non 01 et conditions environnementales dans un site aquacole (Lagune Ebrié, Côte d'Ivoire)

The annual cycle of Vibrio cholerae in the environment surrounding the Layo aquaculture facility (Ebrié lagoon) was studied from March 1991 to April 1992. Vibrio cholerae counts were coupled with the determination of physical and chemical characteristics of water and the estimation of biological richness of this environment.

River Lune juvenile fish stock assessment 2002 (annotated draft copy, Site Reports only)

This draft copy of the River Lune juvenile fish stock assessment from 2000 provides "Site Reports" from different water bodies in the Lune catchment. These Site Reports provide brief information on habitat features, fishery classification and comments on species caught and stocking. This document provides no summary or interpretation of the given data.

A nursery site of the Alaska skate (Bathyraja parmifera) in the eastern Bering Sea

A nursery site for the Alaska skate (Bathyraja parmifera) was sampled seasonally from June 2004 to July 2005. At the small nursery site (~2 km2), located in a highly productive area near the shelf-slope interface at the head of Bering Canyon in the eastern Bering Sea, reproductive males and females dominated the catch and neonate and juvenile skates were rare. Seasonal samples showed summertime (June and July) as the peak reproductive time in the nursery although some reproduction occurred throughout the year. Timeseries analysis of embryo length frequencies revealed that three cohorts were developing simultaneously and the period of embryonic development was estimated at 3.5 years and average embryo growth rate at 0.2 mm/day. Estimated egg case deposition occurred mainly during summertime and hatching occurred during winter months. Protracted hatching times may be common for oviparous elasmobranch species and may be directly correlated with ambient temperatures as evident from a meta-data analysis. Evidence indicates that the Alaska skate uses the eastern Bering Sea outer continental shelf region for reproduction and the middle and inner shelf regions as habitat for immature and subadults. Skate nurseries may be vulnerable to disturbances because they are located in highly productive areas and because embryos develop slowly.

Comparing the identification of southern California juvenile rockfishes (genus Sebastes spp.) by restriction site analysis of the mitochondrial ND3/ND4 region and by morphological characteristics

Rockfish (Sebastes spp.) juveniles are often difficult to identify by using morphological characters. This study independently applies morphological characters and a key based on mitochondrial restriction site variation to identify juvenile rockf ishes collected in southern California during juvenile rockfish surveys. Twenty-four specimens of Sebastes were examined genetically without knowledge of the morphological assignment. Seventeen fish were identified genetically as S. semicinctus, S. goodei, S. auriculatus, S. jordani, S. levis, S. rastrelliger, and S. saxicola. Identities for the remaining fish were narrowed to two or three species: 1) three fish were either S. carnatus or S. chrysomelas; 2) one fish was either S. chlorosticus, S. eos, or S. rosenblatti; and 3) three fish could have been either S. hopkinsi or S. ovalis, the latter for which we now have distinguishing mitochondrial markers. The genetic and morphological assignments concurred except for the identity of one fish that could only be narrowed down to S. hopkinsi or S. semicinctus by using morphological characters. Genetics excluded more species from multispecies groupings than did the morphological approach, especially species within the subgenus Sebastomus. Species in the genetically unresolvable groups may be similar because of recent divergence or because of interspecies introgression.

Como viver sem meu Prozac ? Uma análise antropológica dos discursos sobre o consumo de fluoxetina em um site de relacionamento

Neste trabalho enfocaremos o consumo de antidepressivos, buscando questionar a posição mais comumente aceita para explicar o alto consumo dessas substâncias, que geralmente responsabiliza os ardilosos estratagemas das indústrias farmacêuticas ou a hegemonia da psiquiatria biológica na medicina. Acreditamos que as respostas devem ser buscadas em análises mais profundas, e não simplesmente na demonização desse ou daquele ator social considerado isolada,e,tem, pois admitimos que no consumo se constrói parte da racionalidade integrativa e comunicativa de uma sociedade, logo, pensar o consumo implica em um enfoque dos sujeitos enquanto consumidores, indivíduos e cidadãos. Partindo desses considerações, analisaremos o consumo de antidepressivos pela ótica dos usuários através da análise do conteúdo de um site de relacionamentos muito popular na internet brasileira, conhecido como Orkut.