Relationships between hardness, elastic modulus, and the work of indentation

The work done during indentation is examined using dimensional analysis and finite element calculations for conical indentation in elastic-plastic solids with work hardening. An approximate relationship between the ratio of hardness to elastic modulus and the ratio of irreversible work to total work in indentation is found. Consequently, the ratio of hardness to elastic modulus may be obtained directly from measuring the work of indentation. Together with a well-known relationship between elastic modulus, initial unloading slope, and contact area, a new method is then suggested for estimating the hardness and modulus of solids using instrumented indentation with conical or pyramidal indenters.

Scaling, Dimensional Analysis, and Indentation Measurements

We provide an overview of the basic concepts of scaling and dimensional analysis, followed by a review of some of the recent work on applying these concepts to modeling instrumented indentation measurements. Specifically, we examine conical and pyramidal indentation in elastic-plastic solids with power-law work-hardening, in power-law creep solids, and in linear viscoelastic materials. We show that the scaling approach to indentation modeling provides new insights into several basic questions in instrumented indentation, including, what information is contained in the indentation load-displacement curves? How does hardness depend on the mechanical properties and indenter geometry? What are the factors determining piling-up and sinking-in of surface profiles around indents? Can stress-strain relationships be obtained from indentation load-displacement curves? How to measure time dependent mechanical properties from indentation? How to detect or confirm indentation size effects? The scaling approach also helps organize knowledge and provides a framework for bridging micro- and macroscales. We hope that this review will accomplish two purposes: (1) introducing the basic concepts of scaling and dimensional analysis to materials scientists and engineers, and (2) providing a better understanding of instrumented indentation measurements.

GABA release by hippocampal astrocytes

10 p.

Der Fischei-Test: Ein Toxizitätstest für ökotoxikologische Untersuchungen

Zebrafish (Danio rerio) embryos have been used to quantify the teratogenic potential of environmental samples and harmful substances respectively. The short spawning interval renders this species a good test organism in toxicological research. Due to the transparency of the eggs several lethal and non-lethal endpoints can be detected in parallel after 48 h of embryonic development. Zebrafishembryos have been shown to be sensitive to a number of environmental relevant contaminants, as well as to ex-tracts from polluted sediments

Recognition of double helical DNA by purine oligonucleotides via triple helix formation

Oligonucleotide-directed triple helix formation is one of the most versatile methods for the sequence specific recognition of double helical DNA. Chapter 2 describes affinity cleaving experiments carried out to assess the recognition potential for purine-rich oligonucleotides via the formation of triple helices. Purine-rich oligodeoxyribonucleotides were shown to bind specifically to purine tracts of double helical DNA in the major groove antiparallel to the purine strand of the duplex. Specificity was derived from the formation of reverse Hoogsteen G•GC, A•AT and T•AT triplets and binding was limited to mostly purine tracts. This triple helical structure was stabilized by multivalent cations, destabilized by high concentrations of monovalent cations and was insensitive to pH. A single mismatched base triplet was shown to destabilize a 15 mer triple helix by 1.0 kcal/mole at 25°C. In addition, stability appeared to be correlated to the number of G•GC triplets formed in the triple helix. This structure provides an additional framework as a basis for the design of new sequence specific DNA binding molecules.

In work described in Chapter 3, the triplet specificities and required strand orientations of two classes of DNA triple helices were combined to target double helical sequences containing all four base pairs by alternate strand triple helix formation. This allowed for the use of oligonucleotides containing only natural 3'-5' phosphodiester linkages to simultaneously bind both strands of double helical DNA in the major groove. The stabilities and structures of these alternate strand triple helices depended on whether the binding site sequence was 5'-(purine)_m (pyrimidine)_n-3' or 5'- (pyrimidine)_m (purine)_n-3'.

In Chapter 4, the ability of oligonucleotide-cerium(III) chelates to direct the transesterfication of RNA was investigated. Procedures were developed for the modification of DNA and RNA oligonucleotides with a hexadentate Schiff-base macrocyclic cerium(III) complex. In addition, oligoribonucleotides modified by covalent attachment of the metal complex through two different linker structures were prepared. The ability of these structures to direct transesterification to specific RNA phosphodiesters was assessed by gel electrophoresis. No reproducible cleavage of the RNA strand consistent with transesterification could be detected in any of these experiments.

Biophysical and network mechanisms of high frequency extracellular potentials in the rat hippocampus

A fundamental question in neuroscience is how distributed networks of neurons communicate and coordinate dynamically and specifically. Several models propose that oscillating local networks can transiently couple to each other through phase-locked firing. Coherent local field potentials (LFP) between synaptically connected regions is often presented as evidence for such coupling. The physiological correlates of LFP signals depend on many anatomical and physiological factors, however, and how the underlying neural processes collectively generate features of different spatiotemporal scales is poorly understood. High frequency oscillations in the hippocampus, including gamma rhythms (30-100 Hz) that are organized by the theta oscillations (5-10 Hz) during active exploration and REM sleep, as well as sharp wave-ripples (SWRs, 140-200 Hz) during immobility or slow wave sleep, have each been associated with various aspects of learning and memory. Deciphering their physiology and functional consequences is crucial to understanding the operation of the hippocampal network.

We investigated the origins and coordination of high frequency LFPs in the hippocampo-entorhinal network using both biophysical models and analyses of large-scale recordings in behaving and sleeping rats. We found that the synchronization of pyramidal cell spikes substantially shapes, or even dominates, the electrical signature of SWRs in area CA1 of the hippocampus. The precise mechanisms coordinating this synchrony are still unresolved, but they appear to also affect CA1 activity during theta oscillations. The input to CA1, which often arrives in the form of gamma-frequency waves of activity from area CA3 and layer 3 of entorhinal cortex (EC3), did not strongly influence the timing of CA1 pyramidal cells. Rather, our data are more consistent with local network interactions governing pyramidal cells' spike timing during the integration of their inputs. Furthermore, the relative timing of input from EC3 and CA3 during the theta cycle matched that found in previous work to engage mechanisms for synapse modification and active dendritic processes. Our work demonstrates how local networks interact with upstream inputs to generate a coordinated hippocampal output during behavior and sleep, in the form of theta-gamma coupling and SWRs.

Aplicação da calibração multivariada de ordem superior na resolução de espectros de fluorescência molecular para a quantificação de levofloxacino

Levofloxacino é uma fluorquinolona sintética de 3 geração. É eficaz contra uma variedade de infecções, incluindo o trato respiratório superior e inferior, trato urinário, obstétrico, ginecológico, e infecções dermatológicas. Com o objetivo de quantificar o levofloxacino em medicamentos e amostras de pacientes saudáveis e ter a resolução de seu espectro, foram realizados estudos preliminares em medicamento utilizando espectrofluorescência molecular com concentrações na faixa de 28,8 108 ng/mL e cromatografia líquida de alta eficiência (HPLC) na faixa de concentração de 2,9 10,8 g/mL; e também quantificação em urina de paciente em tratamento com o medicamento, usando os dois métodos citados. Após isso, foram feitos estudos conclusivos utilizando espectrofluorescência molecular e os métodos univariado e PLS para determinação de levofloxacino na faixa de concentração de 0 250 ng/mL e PARAFAC combinado com o método da adição de padrão, para quantificação de levofloxacino em urina de paciente saudável, na faixa de concentração de 0 150 ng/mL, com diluição da amostra em três níveis (100 x, 500 x e 1000x). O método de ordem zero se mostrou mais eficiente na determinação de levofloxacino em medicamento que o de primeira ordem, seus desvios padrão foram 2,0% e 7,9%, respectivamente. Já o PARAFAC com o método de adição de padrão apresentou melhores resultados com a urina, pois possibilitou a quantificação do antibiótico em uma amostra complexa, de forma mais precisa e exata com o aumento da diluição da urina, sem necessidade de tratamento prévio.

Plankton associated with medusae of the freshwater jellyfish Craspedacusta sowerbyi (Lankester) in a Thames backwater

Samples of plankton were taken from Broom Water in August 1997 after a sighting of medusae on 11th August. Broom Water is about 8 m wide, and extends 250 m from the main channel of the Thames, above the weir at Teddington. On 11th August medusae were so abundant that it was possible to collect 20 in ten minutes. They were rising to the surface in bright sunlight, then sinking slowly down through the water. Examination of a medusa's tentacles under a microscope revealed the presence of a commensal protozoan, a ciliate Trichodina pediculus. Over 20 species of phytoplankton were found in Broom Water. Most of the species are common and widespread, but it was a surprise to find Errerella bornhemiensis with its characteristic pyramidal colonies, which is a relatively rare species. Zooplankters in Broom Water consisted of Rotifera and Crustacea. Zooplankton is the main food of Craspedacusta and it was found that the crustaceans but not the rotifers did undergo significant changes during the period 11-19th August. The major changes were a big increase in the percentage of cyclopoids, and a marked decrease in Bosmina. This could be because the delicate cuticle of Bosmina is much more susceptible to the stinging cells of the medusae compared with the tougher exoskeleton of the cyclopoid.

Identificação de áreas sob maior risco para leishmaniose visceral na cidade de Teresina, Piauí, Brasil

O propósito desta Tese foi detectar e caracterizar áreas sob alto risco para leishmaniose visceral (LV) e descrever os padrões de ocorrência e difusão da doença, entre os anos de 1993 a 1996 e 2001 a 2006, em Teresina, Piauí, por meio de métodos estatísticos para análise de dados espaciais, sistemas de informações geográficas e imagens de sensoriamento remoto. Os resultados deste estudo são apresentados na forma de três manuscritos. O primeiro usou análise de dados espaciais para identificar as áreas com maior risco de LV na área urbana de Teresina entre 2001 e 2006. Os resultados utilizando razão de kernels demonstraram que as regiões periféricas da cidade foram mais fortemente afetadas ao longo do período analisado. A análise com indicadores locais de autocorrelação espacial mostrou que, no início do período de estudo, os agregados de alta incidência de LV localizavam-se principalmente na região sul e nordeste da cidade, mas nos anos seguintes os eles apareceram também na região norte da cidade, sugerindo que o padrão de ocorrência de LV não é estático e a doença pode se espalhar ocasionalmente para outras áreas do município. O segundo estudo teve como objetivo caracterizar e predizer territórios de alto risco para ocorrência da LV em Teresina, com base em indicadores socioeconômicos e dados ambientais, obtidos por sensoriamento remoto. Os resultados da classificação orientada a objeto apontam a expansão da área urbana para a periferia da cidade, onde antes havia maior cobertura de vegetação. O modelo desenvolvido foi capaz de discriminar 15 conjuntos de setores censitário (SC) com diferentes probabilidades de conterem SC com alto risco de ocorrência de LV. O subconjunto com maior probabilidade de conter SC com alto risco de LV (92%) englobou SC com percentual de chefes de família alfabetizados menor que a mediana (≤64,2%), com maior área coberta por vegetação densa, com percentual de até 3 moradores por domicílio acima do terceiro quartil (>31,6%). O modelo apresentou, respectivamente, na amostra de treinamento e validação, sensibilidade de 79% e 54%, especificidade de 74% e 71%, acurácia global de 75% e 67% e área sob a curva ROC de 83% e 66%. O terceiro manuscrito teve como objetivo avaliar a aplicabilidade da estratégia de classificação orientada a objeto na busca de possíveis indicadores de cobertura do solo relacionados com a ocorrência da LV em meio urbano. Os índices de acurácia foram altos em ambas as imagens (>90%). Na correlação da incidência da LV com os indicadores ambientais verificou-se correlações positivas com os indicadores Vegetação densa, Vegetação rasteira e Solo exposto e negativa com os indicadores Água, Urbana densa e Urbana verde, todos estatisticamente significantes. Os resultados desta tese revelam que a ocorrência da LV na periferia de Teresina está intensamente relacionada às condições socioeconômicas inadequadas e transformações ambientais decorrentes do processo de expansão urbana, favorecendo a ocorrência do vetor (Lutzomyia longipalpis) nestas regiões.

Contribution of the temporoammonic pathway to hippocampal processing

The temporoammonic (TA) pathway is the direct, monosynaptic projection from layer III of entorhinal cortex to the distal dendritic region of area CA1 of the hippo­ campus. Although this pathway has been implicated in various functions, such as memory encoding and retrieval, spatial navigation, generation of oscillatory activity, and control of hippocampal excitability, the details of its physiology are not well understood. In this thesis, I examine the contribution of the TA pathway to hippocampal processing. I find that, as has been previously reported, the TA pathway includes both excitatory, glutamatergic components and inhibitory, GABAergic components. Several new discoveries are reported in this thesis. I show that the TA pathway is subject to forms of short-term activity-dependent regulation, including paired-pulse and frequency­ dependent plasticity, similar to other hippocampal pathways such as the Schaffer collateral (SC) input from CA3 to CA1. The TA pathway provides a strongly excitatory input to stratum radiatum giant cells of CA1. The excitatory component of the TA pathway undergoes a long-lasting decrease in synaptic strength following low-frequency stimulation in a manner partially dependent on the activation of NMDA receptors. High­ frequency activation of the TA pathway recruits a feedforward inhibition that can prevent CA1 pyramidal cells from spiking in response to SC input; this spike-blocking effect shows that the TA pathway can act to regulate information flow through the hippocampal trisynaptic pathway.

Electronic structure in five-coordinate complexes of nickel(II) with heavy-donor ligands

I.

Various studies designed to elucidate the electronic structure of the arsenic donor ligand, o-phenylenebisdimethylarsine (diarsine), have been carried out. The electronic spectrum of diarsine has been measured at 300 and 77˚K. Electronic spectra of the molecular complexes of various substituted organoarsines and phosphines with tetracyanoethylene have been measured and used to estimate the relative ionization potentials of these molecules.

Uv photolysis of arsines in frozen solution (96˚K) has yielded thermally labile, paramagnetic products. These include the molecular cations of the photolyzed compounds. The species (diars)⁺ exhibits hyper-fine splitting due to two equivalent ⁷⁵As(I=3/2) nuclei. Resonances due to secondary products are reported and assignments discussed.

Evidence is presented for the involvement of d-orbitals in the bonding of arsines. In (diars)⁺ there is mixing of arsenic “lone-pair” orbitals with benzene ring π-orbitals.

II.

Detailed electronic spectral measurements at 300 and 77˚K have been carried out on five-coordinate complexes of low-spin nickel(II), including complexes of both trigonal bipyramidal (TBP) and square pyramidal (SPY) geometry. TBP complexes are of the form NiLX⁺ (X=halide or cyanide,

L = Qƭ(CH₂)₃As(CH₃)₂]₃ or

P [hexagon - Q'CH₃] , Q = P, As,

Q’=S, Se).

The electronic spectra of these compounds exhibit a novel feature at low temperature. The first ligand field band, which is asymmetric in the room temperature solution spectrum, is considerably more symmetrical at 77˚K. This effect is interpreted in terms of changes in the structure of the complex.

The SPY complexes are of the form Ni(diars)₂X^z (X=CL, Br, CNS, CN, thiourea, NO₂, As). On the basis of the spectral results, the d-level ordering is concluded to be xy ˂ xz, yz ˂ z² ˂˂ x² - y². Central to this interpretation is identification of the symmetry-allowed ¹A₁ → ¹E (xz, yz → x² - y²) transition. This assignment was facilitated by the low temperature measurements.

An assignment of the charge-transfer spectra of the five-coordinate complexes is reported, and electronic spectral criteria for distinguishing the two limiting geometries are discussed.

Biochemical and Biophysical Characterization of Huntingtin

Huntington’s disease (HD) is a fatal autosomal dominant neurodegenerative disease. HD has no cure, and patients pass away 10-20 years after the onset of symptoms. The causal mutation for HD is a trinucleotide repeat expansion in exon 1 of the huntingtin gene that leads to a polyglutamine (polyQ) repeat expansion in the N-terminal region of the huntingtin protein. Interestingly, there is a threshold of 37 polyQ repeats under which little or no disease exists; and above which, patients invariably show symptoms of HD. The huntingtin protein is a 350 kDa protein with unclear function. As the polyQ stretch expands, its propensity to aggregate increases with polyQ length. Models for polyQ toxicity include formation of aggregates that recruit and sequester essential cellular proteins, or altered function producing improper interactions between mutant huntingtin and other proteins. In both models, soluble expanded polyQ may be an intermediate state that can be targeted by potential therapeutics.

In the first study described herein, the conformation of soluble, expanded polyQ was determined to be linear and extended using equilibrium gel filtration and small-angle X-ray scattering. While attempts to purify and crystallize domains of the huntingtin protein were unsuccessful, the aggregation of huntingtin exon 1 was investigated using other biochemical techniques including dynamic light scattering, turbidity analysis, Congo red staining, and thioflavin T fluorescence. Chapter 4 describes crystallization experiments sent to the International Space Station and determination of the X-ray crystal structure of the anti-polyQ Fab MW1. In the final study, multimeric fibronectin type III (FN3) domain proteins were engineered to bind with high avidity to expanded polyQ tracts in mutant huntingtin exon 1. Surface plasmon resonance was used to observe binding of monomeric and multimeric FN3 proteins with huntingtin.

Proton-Coupled Reduction of N2 Facilitated by Molecular Fe Complexes

The activation of Fe-coordinated N₂ via the formal addition of hydrogen atom equivalents is explored in this thesis. These reactions may occur in nitrogenase enzymes during the biological conversion of N₂ to NH₃. To understand these reactions, the N₂ reactivity of a series of molecular Fe(N₂) platforms is investigated. A trigonal pyramidal, carbon-ligated Fe^I complex was prepared that displays a similar geometry to that of the resting state 'belt' Fe atoms of nitrogenase. Upon reduction, this species was shown to coordinate N₂, concomitant with significant weakening of the C-Fe interaction. This hemilability of the axial ligand may play a critical role in mediating the interconversion of Fe(N_xH_y) species during N₂ conversion to NH₃. In fact, a trigonal pyramidal borane-ligated Fe complex was shown to catalyze this transformation, generating up to 8.49 equivalents of NH₃. To shed light on the mechanistic details of this reaction, protonation of a borane-ligated Fe(N₂) complex was investigated and found to give rise to a mixture of species that contains an iron hydrazido(2-) [Fe(NNH₂)] complex. The identification of this species is suggestive of an early N-N bond cleavage event en route to NH₃ production, but the highly-reactive nature of this complex frustrated direct attempts to probe this possibility. A structurally-analogous silyl-ligated Fe(N₂) complex was found to react productively with hydrogen atom equivalents, giving rise to an isolable Fe(NNH₂) species. Spectroscopic and crystallographic studies benefited from the enhanced stability of this complex relative to the borane analogue. One-electron reduction of this species initiates a spontaneous disproportionation reaction with an iron hydrazine [Fe(NH₂NH₂)] complex as the predominant reaction product. This transformation provides support for an Fe-mediated N₂ activation mechanism that proceeds via a late N-N bond cleavage. In hopes of gaining more fundamental insight into these reactions, a series of Fe(CN) complexes were prepared and reacted with hydrogen-atom equivalents. Significant quantities of CH₄ and NH₃ are generated in these reactions as a result of complete C-N bond activation. A series of Fe(CNH_x) were found to be exceptionally stable and may be intermediates in these reactions. The stability of these compounds permitted collection of thermodynamic parameters pertinent to the unique N-H bonds. This data is comparatively discussed with the theoretically-predicted data of the N₂-derived Fe(NNH_x) species. Exceptionally-weak N-H bond enthalpies are found for many of these compounds, and sheds light on their short-lived nature and tendency to evolve H₂. As a whole, these works both establish and provide a means to understand Fe-mediated N₂ activation via the addition of hydrogen atom equivalents.

A Continuum Model for Slip-Twinning Interactions in Magnesium and Magnesium Alloys

Due to their high specific strength and low density, magnesium and magnesium-based alloys have gained great technological importance in recent years. However, their underlying hexagonal crystal structure furnishes Mg and its alloys with a complex mechanical behavior because of their comparably smaller number of energetically favorable slip systems. Besides the commonly studied slip mechanism, another way to accomplish general deformation is through the additional mechanism of deformation-induced twinning. The main aim of this thesis research is to develop an efficient continuum model to understand and ultimately predict the material response resulting from the interaction between these two mechanisms.

The constitutive model we present is based on variational constitutive updates of plastic slips and twin volume fractions and accounts for the related lattice reorientation mechanisms. The model is applied to single- and polycrystalline pure magnesium. We outline the finite-deformation plasticity model combining basal, pyramidal, and prismatic dislocation activity as well as a convexification based approach for deformation twinning. A comparison with experimental data from single-crystal tension-compression experiments validates the model and serves for parameter identification. The extension to polycrystals via both Taylor-type modeling and finite element simulations shows a characteristic stress-strain response that agrees well with experimental observations for polycrystalline magnesium. The presented continuum model does not aim to represent the full details of individual twin-dislocation interactions, yet it is sufficiently efficient to allow for finite element simulations while qualitatively capturing the underlying microstructural deformation mechanisms.