Integration of technologies for understanding the functional relationship between reef habitat and fish growth and production

Functional linkage between reef habitat quality and fish growth and production has remained elusive. Most current research is focused on correlative relationships between a general habitat type and presence/absence of a species, an index of species abundance, or species diversity. Such descriptive information largely ignores how reef attributes regulate reef fish abundance (density-dependent habitat selection), trophic interactions, and physiological performance (growth and condition). To determine the functional relationship between habitat quality, fish abundance, trophic interactions, and physiological performance, we are using an experimental reef system in the northeastern Gulf of Mexico where we apply advanced sensor and biochemical technologies. Our study site controls for reef attributes (size, cavity space, and reef mosaics) and focuses on the processes that regulate gag grouper (Mycteroperca microlepis) abundance, behavior and performance (growth and condition), and the availability of their pelagic prey. We combine mobile and fixed-active (fisheries) acoustics, passive acoustics, video cameras, and advanced biochemical techniques. Fisheries acoustics quantifies the abundance of pelagic prey fishes associated with the reefs and their behavior. Passive acoustics and video allow direct observation of gag and prey fish behavior and the acoustic environment, and provide a direct visual for the interpretation of fixed fisheries acoustics measurements. New application of biochemical techniques, such as Electron Transport System (ETS) assay, allow the in situ measurement of metabolic expenditure of gag and relates this back to reef attributes, gag behavior, and prey fish availability. Here, we provide an overview of our integrated technological approach for understanding and quantifying the functional relationship between reef habitat quality and one element of production – gag grouper growth on shallow coastal reefs.

Potential applications of reproductive and molecular genetic technologies in the selective breeding of aquaculture species

The use of reproductive and genetic technologies can increase the efficiency of selective breeding programs for aquaculture species. Four technologies are considered, namely: marker-assisted selection, DNA fingerprinting, in-vitro fertilization, and cryopreservation. Marker-assisted selection can result in greater genetic gain, particularly for traits difficult or expensive to measure, than conventional selection methods, but its application is currently limited by lack of high density linkage maps and by the high cost of genotyping. DNA fingerprinting is most useful for genetic tagging and parentage verification. Both in-vitro fertilization and cryopreservation techniques can increase the accuracy of selection while controlling accumulation of inbreeding in long-term selection programs. Currently, the cost associated with the utilization of reproductive and genetic techniques is possibly the most important factor limiting their use in genetic improvement programs for aquatic species.

The improved management of small-scale cage culture in Asia: final technical report

The purpose of the project is to develop sustained small-scale cage fish culture in inland and coastal waters through improved understanding of the social, institutional and resource environment of resource poor groups. Two Asian countries, Bangladesh (inland systems) and Vietnam (marine), were studied with this workshop bringing together both sides of the project together with representatives of collaborative institutions, government departments and universities. Addressing the overall aim of producing guidelines for the planning and extension of cage aquaculture in Asia a combination of group work and plenary discussion was conducted producing the following outputs. 1) An assessment of cage aquaculture potential, 2) Development options for small-scale cage culture, 3) A review of tools and methodologies and 4) Policy initiatives for sustainable cage culture development. Key issues raised were the use of outputs as a guide to be adapted to regional circumstances to facilitate farmer and extension worker discussion and not as a rigid methodology. The degree of linkage between development, research and government institutions was also considered a crucial factor in benefiting the research and development of cage culture at the local, regional and national level and vital in affecting the future policies by both development and government institutions. [PDF contains 242 pages]

Conservation needs of fisheries resources and reorientation for sustainable captive and culture practices

A discussion is presented on the importance of conservation of fishery resources, considering in particular the situation in Nigeria and providing also an outline of various legislations made by government. Suggestions are given regarding the following: education on the importance of conservation; fish farming or aquaculture; restocking or rejuvenation of depleted water bodies; stock assessment; monitoring and surveillance; provision of fishing inputs; and, extension research linkage

If the tide is rising, who pays for the ark?

Two common goals of this meeting are to arrest the effects of sea level rise and other phenomena caused by Greenhouse Gases from anthropogenic sources ("GHG",) and to mitigate the effects. The fundamental questions are: (1) how to get there and (2) who should shoulder the cost? Given Washington gridlock, states, NGO's and citizens such as the Inupiat of the Village of Kivalina have turned to the courts for solutions. Current actions for public nuisance seek (1) to reduce and eventually eliminate GHG emissions, (2) damages for health effects and property damage—plus hundreds of millions in dollars spent to prepare for the foregoing. The U.S. Court of Appeals just upheld the action against the generators of some 10% of the CO2 emissions from human activities in the U.S., clearing the way for a trial featuring the state of the art scientific linkage between GHG production and the effects of global warming. Climate change impacts on coastal regions manifest most prominently through sea level rise and its impacts: beach erosion, loss of private and public structures, relocation costs, loss of use and accompanying revenues (e.g. tourism), beach replenishment and armoring costs, impacts of flooding during high water events, and loss of tax base. Other effects may include enhanced storm frequency and intensity, increased insurance risks and costs, impacts to water supplies, fires and biological changes through invasions or local extinctions (IPCC AR4, 2007; Okmyung, et al., 2007). There is an increasing urgency for federal and state governments to focus on the local and regional levels and consistently provide the information, tools, and methods necessary for adaptation. Calls for action at all levels acknowledge that a viable response must engage federal, state and local expertise, perspectives, and resources in a coordinated and collaborative effort. A workshop held in December 2000 on coastal inundation and sea level rise proposes a shared framework that can help guide where investments should be made to enable states and local governments to assess impacts and initiate adaptation strategies over the next decade. (PDF contains 5 pages)

A Convergent Synthetic Route to the Tunicamycin Antiobiotics. Synthesis of (+)-Tunicamycin V

A concise synthetic route to the tunicamycin antibiotics is described, illustrated by the preparation of (+)-tunicamycin-V (1-V). Key features of the synthesis include: (1) the development and application of a silicon-mediated reductive coupling of aldehydes and allylic alcohols to construct the undecose core of the natural product; and (2) the development of an efficient procedure for the synthesis of the trehalose glycosidic bond within the antibiotic. These innovations allow for the coupling of a uridine-derived aldehyde fragment with a preformed trehalose-linked disaccharide allylic alcohol to form the carbohydrate core (1) of the natural product in a highly convergent manner. The resultant amino polyol is a versatile intermediate for the synthesis of any of the homologous tunicamycin antibiotics.

Multiplexed DNA-mediated electrochemistry

The aromatic core of double helical DNA possesses the unique and remarkable ability to form a conduit for electrons to travel over exceptionally long molecular distances. This core of π-stacked nucleobases creates an efficient pathway for charge transfer to proceed that is exquisitely sensitive to even subtle perturbations. Ground state electrochemistry of DNA-modified electrodes has been one of the major techniques used both to investigate and to harness the property of DNA-mediated charge transfer. DNA-modified electrodes have been an essential tool for both gaining insights into the fundamental properties of DNA and, due to the exquisite specificity of DNA-mediated charge transfer for the integrity of the π-stack, for use in next generation diagnostic sensing. Here, multiplexed DNA-modified electrodes are used to (i) gain new insights on the electrochemical coupling of metalloproteins to the DNA π-stack with relevance to the fundaments of in vivo DNA-mediated charge transfer and (ii) enhance the overall sensitivity of DNA-mediated reduction for use in the detection of low abundance diagnostic targets.

First, Methylene Blue (MB′) was covalently attached to DNA through a flexible C12 alkyl linker to yield a new redox reporter for DNA electrochemistry measurements with enhanced sensitivity. Tethered, intercalated MB′ was reduced through DNA-mediated charge transport. The redox signal intensity for MB′-dT-C12-DNA was found to be at least 3 fold larger than that of previously used Nile Blue (NB)-dT-DNA, which is coupled to the base stack via direct conjugation. The signal attenuation, due to an intervening mismatch, and therefore the degree of DNA-mediated reduction, does, however, depend on the DNA film morphology and the backfilling agent used to passivate the surface. These results highlight two possible mechanisms for the reduction of MB′ on the DNA-modified electrode that are distinguishable by their kinetics: reduction mediated by the DNA base pair stack and direct surface reduction of MB′ at the electrode. The extent of direct reduction at the surface can be minimized by overall DNA assembly conditions.

Next, a series of intercalation-based DNA-mediated electrochemical reporters were developed, using a flexible alkane linkage to validate and explore their DNA-mediated reduction. The general mechanism for the reduction of distally bound redox active species, covalently tethered to DNA through flexible alkyl linkages, was established to be an intraduplex DNA-mediated pathway. MB, NB, and anthraquinone were covalently tethered to DNA with three different covalent linkages. The extent of electronic coupling of the reporter was shown to correlate with the DNA binding affinity of the redox active species, supporting an intercalative mechanism. These electrochemical signals were shown to be exceptionally sensitive to a single intervening π-stack perturbation, an AC mismatch, in a densely packed DNA monolayer, which further supports that the reduction is DNA-mediated. Finally, this DNA-mediated reduction of MB occurs primarily via intra- rather than inter duplex intercalation, as probed through varying the proximity and integrity of the neighboring duplex DNA. Further gains to electrochemical sensitivity of our DNA-modified devices were then achieved through the application of electrocatalytic signal amplification using these solvent accessible intercalative reporters, MB-dT-C8, and hemoglobin as a novel electron sink. Electrocatalysis offers an excellent means of electrochemical signal amplification, yet in DNA based sensors, its application has been limited due to strict assembly conditions. We describe the use of hemoglobin as a robust and effective electron sink for electrocatalysis in DNA sensing on low density DNA films. Protein shielding of the heme redox center minimizes direct reduction at the electrode surface and permits assays on low density DNA films. Electrocatalysis of MB that is covalently tethered to the DNA by a flexible alkyl linkage allows for efficient interactions with both the base stack and hemoglobin. Consistent suppression of the redox signal upon incorporation of single CA mismatch in the DNA oligomer demonstrates that both the unamplified and the electrocatalytically amplified redox signals are generated through DNA-mediated charge transport. Electrocatalysis with hemoglobin is robust: it is stable to pH and temperature variations. The utility and applicability of electrocatalysis with hemoglobin is demonstrated through restriction enzyme detection, and an enhancement in sensitivity permits femtomole DNA sampling.

Finally, we expanded the application of our multiplexed DNA-modified electrodes to the electrochemical characterization of DNA-bound proteins containing [4Fe-4S] clusters. DNA-modified electrodes have become an essential tool for the characterization of the redox chemistry of DNA repair proteins that contain redox cofactors. Multiplexed analysis of EndonucleaseIII (EndoIII), a DNA repair protein containing a [4Fe-4S] cluster known to be accessible via DNA-mediated charge transport, elucidated subtle differences in the electrochemical behavior as a function of DNA morphology. DNA-bound EndoIII is seen to have two different electron transfer pathways for reduction, either through the DNA base stack or through direct surface reduction. Closely packed DNA films, where the protein has limited surface accessibility, produce electrochemical signals reflecting electron transfer that is DNA-mediated. The electrochemical comparison of EndoIII mutants, including a new family of mutations altering the electrostatics surrounding the [4Fe-4S] cluster, was able to be quantitatively performed. While little change in the midpoint potential was found for this family of mutants, significant variations in the efficiency of DNA-mediated electron transfer were apparent. Based on the stability of these proteins, examined by circular dichroism, we propose that the electron transfer pathway can be perturbed not only by the removal of aromatic residues, but also through changes in solvation near the cluster.

Electron tunneling in proteins and water

The subject of this thesis is electronic coupling in donor-bridge-acceptor systems. In Chapter 2, ET properties of cyanide-bridged dinuclear ruthenium complexes were investigated. The strong interaction between the mixed-valent ruthenium centers leads to intense metal-to-metal charge transfer bands (MMCT). Hush analysis of the MMCT absorption bands yields the electronic-coupling strength between the metal centers (H_(AB)) and the total reorganization energy (λ). Comparison of ET kinetics to calculated rates shows that classical ET models fail to account for the observed kinetics and nuclear tunneling must be considered.

In Chapter 3, ET rates were measured in four ruthenium-modified highpotential iron-sulfur proteins (HiPIP), which were modified at position His50, His81, His42 and His18, respectively. ET kinetics for the His50 and His81 mutants are a factor of 300 different, while the donor-acceptor separation is nearly identical. PATHWAY calculations corroborate these measurements and highlight the importance of structural detail of the intervening protein matrix.

In Chapter 4, the distance dependence of ET through water bridges was measured. Photoinduced ET measurements in aqueous glasses at 77 K show that water is a poor medium for ET. Luminescence decay and quantum yield data were analyzed in the context of a quenching model that accounts for the exponential distance dependence of ET, the distance distribution of donors and acceptors embedded in the glass and the excluded volumes generated by the finite sizes of the donors and acceptors.

In Chapter 5, the pH-dependent excited state dynamics of ruthenium-modified amino acids were measured. The [Ru(bpy)_(3)] ^(2+) chromophore was linked to amino acids via an amide linkage. Protonation of the amide oxygen effectively quenches the excited state. In addition. time-resolved and steady-state luminescence data reveal that nonradiative rates are very sensitive to the protonation state and the structure of the amino acid moiety.

Part I. Synthesis of L-amino acid oxidase by a serine- or glycine-requiring strain of neurospora. Part II. Studies concerning multiple electrophoretic forms of tyrosinase in neurospora

Part I: Synthesis of L-Amino Acid Oxidase by a Serine- or Glycine-Requiring Strain of Neurospora

Wild-type cultures of Neurospora crassa growing on minimal medium contain low levels of L-amino acid oxidase, tyrosinase, and nicotinarnide adenine dinucleotide glycohydrase (NADase). The enzymes are derepressed by starvation and by a number of other conditions which are inhibitory to growth. L-amino acid oxidase is, in addition, induced by growth on amino acids. A mutant which produces large quantities of both L-amino acid oxidase and NADase when growing on minimal medium was investigated. Constitutive synthesis of L-amino acid oxidase was shown to be inherited as a single gene, called P110, which is separable from constitutive synthesis of NADase. P110 maps near the centromere on linkage group IV.

L-amino acid oxidase produced constitutively by P110 was partially purified and compared to partially purified L-amino acid oxidase produced by derepressed wild-type cultures. The enzymes are identical with respect to thermostability and molecular weight as judged by gel filtration.

The mutant P110 was shown to be an incompletely blocked auxotroph which requires serine or glycine. None of the enzymes involved in the synthesis of serine from 3-phosphoglyceric acid or glyceric acid was found to be deficient in the mutant, however. An investigation of the free intracellular amino acid pools of P110 indicated that the mutant is deficient in serine, glycine, and alanine, and accumulates threonine and homoserine.

The relationship between the amino acid requirement of P110 and its synthesis of L-amino acid oxidase is discussed.

Part II: Studies Concerning Multiple Electrophoretic Forms of Tyrosinase in Neurospora

Supernumerary bands shown by some crude tyrosinase preparations in paper electrophoresis were investigated. Genetic analysis indicated that the location of the extra bands is determined by the particular T allele present. The presence of supernumerary bands varies with the method used to derepress tyrosinase production, and with the duration of derepression. The extra bands are unstable and may convert to the major electrophoretic band, suggesting that they result from modification of a single protein. Attempts to isolate the supernumerary bands by continuous flow paper electrophoresis or density gradient zonal electrophoresis were unsuccessful.

Interpretações de linguagens artísticas em Gerd Bornheim

Este estudo propõe uma leitura histórico-cultural das interpretações de Gerd Bornheim (1929-2002), destacando a temática da linguagem, sobretudo das linguagens artísticas. A partir dessas expressões, as colocações críticas de Bornheim a respeito da estética e filosofia da arte apresentam um panorama dos questionamentos. Nesse sentido, são notáveis em seus trabalhos as reflexões sobre o teatro e a música. A linguagem teatral permite o acesso às outras atividades artísticas (poesia, música, artes plásticas, cinema) de forma livre e aberta. A linguagem musical, em consonância com a teatral, corrobora a pesquisa de Bornheim, que observou o processo criativo, a comunicação, o papel da interpretação (advento da crítica) e as rupturas nas poéticas contemporâneas. Tal itinerário sublinha a pesquisa que Bornheim realizou na França nas décadas de 1950 e 1960-70. O estímulo dessa atmosfera, marcada pelo diálogo entre filosofia, ciências sociais, psicologia, psicanálise, história, antropologia, linguística, comunicação, teatro e música foi decisivo para ele. Esses pontos são importantes para a apreensão do tema da linguagem e sua ambiência histórica, na qual Bornheim revela outras perspectivas de pesquisa. O pano de fundo é a crise da metafísica e os novos parâmetros para se pensar a dialética, a teoria e a prática. O diagnóstico de tal crise estende-se também à estética. Por conseguinte, o entendimento das ideias de Bornheim conduz aos temas da diferença e alteridade na contemporaneidade. Com isso, persegue-se um percurso temático que aborda: a linguagem e o problema da comunicação a partir da ligação das interpretações de Bornheim com as de Sartre e Merleau-Ponty. Além disso, o surgimento da crítica e os questionamentos da normatividade ética e estética levam à discussão das motivações coincidentes entre artes e ciências. Por fim, a linguagem musical enfatiza ainda o processo de transformação da subjetividade, que propicia uma percepção mais ampla das expressões artísticas e culturais.

A imprescritibilidade da negatória de paternidade e o melhor interesse da criança

Estudo sobre a filiação com ênfase na regra do caput do artigo 1.601 do Código Civil Brasileiro Lei 10.406/2002, segundo a qual cabe ao marido o direito de contestar a paternidade dos filhos nascidos de sua mulher, sendo tal ação imprescritível. A dissertação tem como objetivo principal demonstrar a inconstitucionalidade do direito perpétuo do pai em desconstituir a paternidade do filho menor nascido em uma relação de casamento à luz do princípio do melhor interesse da criança. Para tanto foi realizada pesquisa bibliográfica e jurisprudencial em tribunais nacionais de segunda instância, no Superior Tribunal de Justiça e no Supremo Tribunal Federal. Os critérios da presunção legal pater is est quem nuptiae demonstrant, da verdade biológica e da posse de estado de filho foram um a um analisados, não havendo entre eles predominância no ordenamento jurídico. O levantamento deste material permitiu a constatação de que a regra supracitada é, pela doutrina majoritária, acriticamente louvada e utilizada nos julgados de conflitos de paternidade. O filho, no entanto, não pode permanecer por toda a vida sujeito à possibilidade do pai desfazer, a qualquer momento, a relação paterno-filial pela ausência do dado biológico por que: (i) o exercício deste direito é mutilador de sua identidade e dignidade humana; (ii) a posse de estado de filho confirma a presunção pater is est; (iii) o estado civil de filho deve se tornar certo e estável em um curto período de tempo. Sustenta-se que é imperioso o afastamento por inconstitucionalidade material da regra do artigo 1.601 do Código Civil e a aplicação analógica do prazo de quatro anos previsto no artigo 1.614 do Código Civil. De lege ferenda propõe-se a elaboração de uma norma que fixe um prazo decadencial para que o marido e pai possa exercer o direito potestativo de negar a paternidade.

The binding and catalytic properties of lysozyme

The binding and catalytic properties of hen's egg white lysozyme have been studied by a variety of techniques. These studies show that the enzyme has three contiguous binding subsites, A, B, and C. The application of nuclear magnetic resonance (NMR) spectroscopy to probe the binding environment of several saccharides to lysozyme has demonstrated that the reducing end sugar rings of chitotriose, chitobiose and the β-form of N-acetylglucosamine all bind in subsite C. The central sugar ring of chitotriose and the sugar ring at the nonreducing end of chitobiose were found to bind in subsite B, while the nonreducing end sugar residue of chitotriose occupied subsite A. The dynamics of the binding process has also been investigated by NMR. The formation rate constant of chitobiose--and chitotriose-enzyme complexes were found to be about 4 X 10^-6 M^-1 sec^-1 with small activation energies.

The stereochemical path of the lysozyme catalyzed hydrolysis of glycosidic bonds has been shown to proceed with at least 99.7% retention of configuration at C-1 of the sugar. The lysozyme catalyzed hydrolysis of glucosidic bonds has been shown to be largely carbonium ion in character by virtue of the α-deuterium kinetic isotope effect (k_H/k_D = 1.11) observed for the reaction. It is probable that the mechanism of action of the enzyme involves a carbonium ion intermediate which is stereospecifically quenched by solvent. However, acetamido group participation cannot be ruled out for natural substrates.

Desenvolvimento e validação de sistema multiplex X-STR para identificação humana por DNA

Novas metodologias de análise molecular voltadas para estudos populacionais, clínicos, evolutivos, da biodiversidade e identificação forense foram desenvolvidas com base em marcadores microssátelites ou STR Short Tandem Repeats. Os marcadores STR, que estão amplamente espalhados nos genomas e se caracterizam por apresentar alto grau de polimorfismo, podem ser analisados a partir da amplificação por PCR (Reação em Cadeia da polimerase). A análise foi facilitada a partir do desenvolvimento de sistemas de amplificação simultânea de múltiplos STR (multiplex STR) e com a detecção automatizada dos produtos de amplificação marcados por fluorescência. Recentemente, o uso de marcadores STR do cromossomo X (X-STR) tornou-se significativo na prática forense. Devido ao seu modo de transmissão, os X-STR são úteis em situações particulares de investigação de relações de parentesco, apresentando vantagens sobre o uso de STR autossômicos. Este estudo teve como principal objetivo o desenvolvimento e validação de sistema multiplex, denominado LDD (X-STR) Decaplex, capaz de amplificar dez loci X-STR (DXS7133, DXS7424, DXS8378, DXS6807, DXS7132, DXS10074, DXS7423, DXS8377, GATA172D05 e DXS10101) para aplicação em genética populacional, identificação e análises forenses. Utilizando o LDD (X-STR) Decaplex 170 indivíduos autodenominados afrodescendentes, não aparentados geneticamente, foram genotipados. As freqüências alélicas e genotípicas não apresentaram desvio do equilíbrio de Hardy-Weinberg e estão em concordância com aquelas observadas em outros estudos. Os haplótipos observados foram únicos em indivíduos de amostra masculina. A análise de desequilíbrio de ligação não revelou associação entre os marcadores X-STR. A diversidade genética foi elevada, variando entre 0,6218 para o locus DXS7133 a 0,9327 para o locus DXS8377. Os parâmetros de Probabilidade de Vinculação (PV), Índice de Vinculação (IV), Poder de Exclusão (PE), Poder de Discriminação e Razão de Verossimilhança foram também elevados, demonstraram que os dez X-STRs são altamente polimórficos e discriminativos na população estudada. A concentração mínima de DNA para a amplificação dos loci do LDD (X-STR) Decaplex é de 0,5 ng e verificamos que amplificação por PCR pode ser afetada quando são adicionados mais de 5 ng de DNA nas reações. Os percentuais de bandas stutter foram elevados para os loci DXS7132 e DXS8377. No teste de reprodutibilidade observamos consistência entre as tipagem de diferentes amostras biológicas, incluindo as de restos mortais. No teste de mistura a proporção limite em que observamos a coexistência de duas espécies biológicas foi de 2,5:1ng (feminino-masculino). Os resultados evidenciaram que os loci do LDD (X-STR) Decaplex são altamente informativos, consistindo, em conjunto, uma ferramenta importante em estudos de identificação humana e de relações de parentesco.

Dimorfismo sexual e polimorfismo no gênero Ptychoderes Schoenherr, 1823 (Coleoptera Anthribidae, Anthribinae, Ptychoderini)

O dimorfismo sexual exibido por machos polifênicos em algumas espécies do gênero Ptychoderes envolve variação no rostro, antena e ventritos. A existência de polifenismo pode ser um importante componente no processo evolutivo por meio de novidades morfológicas e comportamentais. O objetivo desse estudo foi determinar a variação em caracteres morfométricos, polifenismo em machos, variação de estruturas com conhecido dimorfismo sexual, possíveis padrões alométricos e testar estas inferências para Ptychoderes através do mapeamento do dimorfismo sexual e de machos em uma reconstrução filogenética de Ptychoderes usando Mesquite 2.04. Foram medidas 23 variáveis morfométricas em 510 espécimes com as seguintes análises realizadas: análises de cluster, analises de componentes principais (PCA), analise de variáveis canônicas (AVC), análise de regressão por eixo maior reduzido (RMA). Cada tipo de dimorfismo foi mapeado em uma filogenia prévia como dois estados separados usando parcimônia. Para todas as espécies o dimorfismo sexual apresentou diferenças significativas entre os sexos com relação aos segmentos antenais (II- X).O compriemtno do rosto e ventrito V foram confirmados como indicativos de dimorfismo sexual (exceto em P. jordani). A única espécie em que não ocorreu machos polifenicos foi P. depressus. Nas outras espécies machos grandes e pequenos diferem significantemente para muitas variáveis com similaridades e diferenças. Na ACP, o primeiro componente (PC1) apresentou alta porcentagem de variância nos dados de todas as espécies; apresentou loadings de mesmo sinal sugerindo diferenças relacionadas ao tamanho para as espécies P. jordani, P. depressus, P. virgatus, P. mixtus e P. callosus e para as espécies P. viridanus, P. antiquus, P. elongates e P. nebulosus apresentou loadings positivos e negativos sugerindo diferenças relacionadas a forma (alometria). O PC2 apresentou loadings positivos e negativos para todas as espécies, um provável componente alométricos. A AVC confirmou os grupos: machos grandes, machos pequenos e fêmeas quando estes ocorreram. Nós encontramos diferentes padrões alométricos para todas as espécies com diferenças e semelhanças entre as espécies. Todos esses resultados confirmam a hipótese de polifenismo em machos e dimorfismo sexual para Ptychhoderes. A análise dos padrões alométricos para o dimorfismo sexual revelou alometria positiva para o comprimento do rostro (CR1) em machos e fêmeas, com os ventritos apenas em machos. Padrões alométricos positivos relacionados ao polifenismo nos antenômeros foram confirmados para os machos grandes e pequenos de quase todas as espécies exceto em P. nebulosus. O ancestral de clados na filogenia de Ptychoderes foi inferido para machos polifênicos (exceto P. depressus) com variáveis no rostro, antenas e ventritos indicativas de dimorfismo sexual com alometria positiva. Estes padrões poderiam estar ligados com o comportamento de proteção das fêemeas realizados por machos grandes durante a oviposição.

Investigação de mutações no gene ATP13A2 como um fator de risco para a Doença de Parkinson em pacientes brasileiros

A doença de Parkinson (DP) é a desordem neurodegenerativa motora mais frequente, com uma prevalência de, aproximadamente, 1% entre indivíduos com mais de 60 anos de idade, aumentando para 4 a 5% entre os indivíduos com idade superior a 85 anos. Esta condição é caracterizada pela perda seletiva dos neurônios dopaminérgicos da substância negra e pela presença de inclusões protéicas ricas em α-sinucleína nos neurônios sobreviventes. Pouco se sabe sobre a etiologia e a patogênese da DP. A maioria dos casos aparece esporadicamente, podendo estar associados a diversos fatores de risco ambientais e genéticos. Na última década, estudos de ligação identificaram 15 loci cromossômicos (PARK1 a PARK15) relacionados à DP e, nestes, um novo gene, ATP13A2, tem sido associado a casos de DP de início precoce. Esse gene está situado no 1p36 e codifica a proteína ATPase tipo-P da subfamília P5, de localização lisossômica, que é expressa em diversos tecidos, principalmente no cérebro. Mutações em ATP13A2 levam à formação de proteínas truncadas que ficam retidas no reticulo endoplasmático e posteriormente são degradadas pelo proteossomo, podendo causar a disfunção proteossômica, decorrente da sobrecarga gerada pela proteína mutante, ou causar a disfunção lisossômica, ambas gerando agregação tóxica. Este trabalho tem como objetivo realizar a análise molecular do gene ATP13A2 em uma amostra de 116 pacientes brasileiros com DP, de manifestação precoce (<50 anos), de forma a avaliar se mutações neste gene representam um fator de risco para a DP. O DNA foi extraído a partir de leucócitos do sangue periférico ou de saliva e a análise molecular dos éxons 2, 3, 12, 13, 14, 15, 16, 26 e 27, bem como, dos limites íntronéxons foi realizada por sequenciamento automático dos produtos da PCR. Identificamos oito variantes de sequência: quatro variantes intrônicas (uma no íntron 2, uma no íntron 13 e duas no íntron 27) e quatro variantes silenciosas (uma no éxon 3, 16, 26 e 27). Com base em dados da literatura e através de análises in silico e comparação com amostras controle, classificamos a alteração intrônica c.3084- 3C>T, e as alterações silenciosas c.2970G>A e c.3192C>T como não patogênicas; as alterações intrônicas c.106-30G>T, c.1306+42_1306+43 insC e c.3083+24C>T, e as alterações silenciosas c.132A>G e c.1610G>T foram classificadas como provavelmente não patogênicas. Nosso achados corroboram àqueles encontrados em outras populações e indicam que mutações no gene ATP13A2 não são uma causa comum de DP na amostra de pacientes brasileiros analisados. No entanto, se faz necessário estender nossas análises para outras regiões gênicas, a fim de determinar o real papel deste gene na etiologia da DP em nossa população.