Estudo comparativo do desempenho de diferentes membranas oclusivas de Látex Natural no reparo ósseo

Introduction: In dentistry, alveolar bone resorption is a limiting factor in the well being of individuals directly interfering in the stomatognathic system, causing problems in the context of overall health. Aiming to promote biological methods that can stimulate bone regeneration, several biomimetic strategies have been developed by the use of diverse materials possible to the bone matrix, culminating in the development of techniques that promote such repair. Objective: This work is a comparative study of the performance of films made with latex as occlusive membrane for Guided Bone Regeneration (GBR) procedure in three preparations: Latex preserved in ammonia, produced by Latex rubber clones IAN873 and PR255 polymerized immediately after collection and without use ammonia as a preservative. Methods: Sixty Wistar rats were randomly divided into 4 groups of 15 animals in which bone defects of critical size (8mm diameter) were made surgically in the skull. Group A was treated by GBR through the membrane latex preserved with ammonia, Group B received the membrane made of latex from IAN873, Group C, the membrane clone PR255 and group D was not treated by GBR. After a period of 7, 15 and 50 days, 5 animals from each group were euthanized, and specimens containing bone defect collected for microscopic examination (descriptive histology and histomorphometry). Results: The results showed that after 50 days there was bone formation in higher proportions in group D (p <0.05, ANOVA followed by Tukey), suggesting that further experiments should be conducted to conclude about the presence of ammonia and the influence of kind of rubber. Conclusion: GOR is a procedure proven effective in the treatment of bone defects. Therefore, further experiments should be conducted to reach a conclusion regarding the presence of ammonia in the latex composition for the manufacture of membranes, as well as the difference induced by the species of rubber.

Prevulcanização do látex natural extraído da hancornia speciosa gomes (Mangabeira) e da hevea brasiliensis (seringueira)

Pós-graduação em Ciência dos Materiais - FEIS

The Perception of Natural, Cell Phone, and Computer-Synthesized Speech During The Performance Of Simultaneous Visual-Motor Tasks

This study investigated the influence of top-down and bottom-up information on speech perception in complex listening environments. Specifically, the effects of listening to different types of processed speech were examined on intelligibility and on simultaneous visual-motor performance. The goal was to extend the generalizability of results in speech perception to environments outside of the laboratory. The effect of bottom-up information was evaluated with natural, cell phone and synthetic speech. The effect of simultaneous tasks was evaluated with concurrent visual-motor and memory tasks. Earlier works on the perception of speech during simultaneous visual-motor tasks have shown inconsistent results (Choi, 2004; Strayer & Johnston, 2001). In the present experiments, two dual-task paradigms were constructed in order to mimic non-laboratory listening environments. In the first two experiments, an auditory word repetition task was the primary task and a visual-motor task was the secondary task. Participants were presented with different kinds of speech in a background of multi-speaker babble and were asked to repeat the last word of every sentence while doing the simultaneous tracking task. Word accuracy and visual-motor task performance were measured. Taken together, the results of Experiments 1 and 2 showed that the intelligibility of natural speech was better than synthetic speech and that synthetic speech was better perceived than cell phone speech. The visual-motor methodology was found to demonstrate independent and supplemental information and provided a better understanding of the entire speech perception process. Experiment 3 was conducted to determine whether the automaticity of the tasks (Schneider & Shiffrin, 1977) helped to explain the results of the first two experiments. It was found that cell phone speech allowed better simultaneous pursuit rotor performance only at low intelligibility levels when participants ignored the listening task. Also, simultaneous task performance improved dramatically for natural speech when intelligibility was good. Overall, it could be concluded that knowledge of intelligibility alone is insufficient to characterize processing of different speech sources. Additional measures such as attentional demands and performance of simultaneous tasks were also important in characterizing the perception of different kinds of speech in complex listening environments.

Natural history of an ant-plant-butterfly interaction in a Neotropical savanna

Non-myrmecophilous lepidopteran larvae using plants bearing ant attractants such as extrafloral nectaries are good models for studying morphological and behavioural mechanisms against ant predation. Udranomia spitzi (Hesperiidae) is a butterfly whose larvae feed on leaves of Ouratea spectabilis (Ochnaceae), a plant with extrafloral nectaries. We described the early stages of U. spitzi, and used field observations and experiments to investigate the defensive strategies of caterpillars against predatory ants. Larvae pass through five instars and pupation occurs inside larval leaf shelters. Ant-exclusion experiments revealed that the presence of ants did not affect significantly caterpillar survival. Predation experiments showed that vulnerability to ant predation decreased with increase in larval size. The present study showed that predatory ants are not as relevant as demonstrated for other systems, and also illustrates how observational data and field experiments can contribute to a better understanding of the biology and ecology of a species of interest.

Bottle effects, toxicity and cell viability during incubation experiments to asses community respiration

Máster en Oceanografía

Natural patterns and anthropogenic disturbance in north Adriatic marine benthic assemblages: descriptive and methodological studies

Marine soft bottom systems show a high variability across multiple spatial and temporal scales. Both natural and anthropogenic sources of disturbance act together in affecting benthic sedimentary characteristics and species distribution. The description of such spatial variability is required to understand the ecological processes behind them. However, in order to have a better estimate of spatial patterns, methods that take into account the complexity of the sedimentary system are required. This PhD thesis aims to give a significant contribution both in improving the methodological approaches to the study of biological variability in soft bottom habitats and in increasing the knowledge of the effect that different process (both natural and anthropogenic) could have on the benthic communities of a large area in the North Adriatic Sea. Beta diversity is a measure of the variability in species composition, and Whittaker’s index has become the most widely used measure of beta-diversity. However, application of the Whittaker index to soft bottom assemblages of the Adriatic Sea highlighted its sensitivity to rare species (species recorded in a single sample). This over-weighting of rare species induces biased estimates of the heterogeneity, thus it becomes difficult to compare assemblages containing a high proportion of rare species. In benthic communities, the unusual large number of rare species is frequently attributed to a combination of sampling errors and insufficient sampling effort. In order to reduce the influence of rare species on the measure of beta diversity, I have developed an alternative index based on simple probabilistic considerations. It turns out that this probability index is an ordinary Michaelis-Menten transformation of Whittaker's index but behaves more favourably when species heterogeneity increases. The suggested index therefore seems appropriate when comparing patterns of complexity in marine benthic assemblages. Although the new index makes an important contribution to the study of biodiversity in sedimentary environment, it remains to be seen which processes, and at what scales, influence benthic patterns. The ability to predict the effects of ecological phenomena on benthic fauna highly depends on both spatial and temporal scales of variation. Once defined, implicitly or explicitly, these scales influence the questions asked, the methodological approaches and the interpretation of results. Problem often arise when representative samples are not taken and results are over-generalized, as can happen when results from small-scale experiments are used for resource planning and management. Such issues, although globally recognized, are far from been resolved in the North Adriatic Sea. This area is potentially affected by both natural (e.g. river inflow, eutrophication) and anthropogenic (e.g. gas extraction, fish-trawling) sources of disturbance. Although few studies in this area aimed at understanding which of these processes mainly affect macrobenthos, these have been conducted at a small spatial scale, as they were designated to examine local changes in benthic communities or particular species. However, in order to better describe all the putative processes occurring in the entire area, a high sampling effort performed at a large spatial scale is required. The sedimentary environment of the western part of the Adriatic Sea was extensively studied in this thesis. I have described, in detail, spatial patterns both in terms of sedimentary characteristics and macrobenthic organisms and have suggested putative processes (natural or of human origin) that might affect the benthic environment of the entire area. In particular I have examined the effect of off shore gas platforms on benthic diversity and tested their effect over a background of natural spatial variability. The results obtained suggest that natural processes in the North Adriatic such as river outflow and euthrophication show an inter-annual variability that might have important consequences on benthic assemblages, affecting for example their spatial pattern moving away from the coast and along a North to South gradient. Depth-related factors, such as food supply, light, temperature and salinity play an important role in explaining large scale benthic spatial variability (i.e., affecting both the abundance patterns and beta diversity). Nonetheless, more locally, effects probably related to an organic enrichment or pollution from Po river input has been observed. All these processes, together with few human-induced sources of variability (e.g. fishing disturbance), have a higher effect on macrofauna distribution than any effect related to the presence of gas platforms. The main effect of gas platforms is restricted mainly to small spatial scales and related to a change in habitat complexity due to a natural dislodgement or structure cleaning of mussels that colonize their legs. The accumulation of mussels on the sediment reasonably affects benthic infauna composition. All the components of the study presented in this thesis highlight the need to carefully consider methodological aspects related to the study of sedimentary habitats. With particular regards to the North Adriatic Sea, a multi-scale analysis along natural and anthopogenic gradients was useful for detecting the influence of all the processes affecting the sedimentary environment. In the future, applying a similar approach may lead to an unambiguous assessment of the state of the benthic community in the North Adriatic Sea. Such assessment may be useful in understanding if any anthropogenic source of disturbance has a negative effect on the marine environment, and if so, planning sustainable strategies for a proper management of the affected area.

Characterization of inclusions and their distribution in natural and artificial samples by synchrotron cryo-micro-tomography (SCXRT)

Zur geometrischen Vermessung und Beschreibung von Einschlüssen in natürlichen sowie im Labor geschaffenen Eispartikeln wurde ein neuartiger Versuchaufbau an der Tomographie-Endstation der Material Science Beam Line an der Swiss Light Source (SLS, Paul Scherrer Institut, Villigen, Schweiz) entwickelt. Dieser besteht aus einer Plexiglas-Tasse und einem doppelwandigen Kaptonfolien-Käfig, der wiederum auf die Düse eines CryojetXL (Oxford Instruments) montiert wurde. Abgesehen von dem hohen Maß an Flexibilit¨at bez¨uglich der Installation erlaubt es dieser Aufbau, die Temperatur des Experiments mit einer Genauigkeit von ± 1 K über einen Bereich von 271 K bis 220 K zu regeln. In den hier beschriebenen Experimenten wurde eine räumliche Auflösung von 1.4 µm erzielt.

Natural compounds Camptothecin and Triptolide: highly specific enzyme inhibitors and tools to dissect transcriptional functions

With this work I elucidated new and unexpected mechanisms of two strong and highly specific transcription inhibitors: Triptolide and Campthotecin. Triptolide (TPL) is a diterpene epoxide derived from the Chinese plant Trypterigium Wilfoordii Hook F. TPL inhibits the ATPase activity of XPB, a subunit of the general transcription factor TFIIH. In this thesis I found that degradation of Rbp1 (the largest subunit of RNA Polymerase II) caused by TPL treatments, is preceded by an hyperphosphorylation event at serine 5 of the carboxy-terminal domain (CTD) of Rbp1. This event is concomitant with a block of RNA Polymerase II at promoters of active genes. The enzyme responsible for Ser5 hyperphosphorylation event is CDK7. Notably, CDK7 downregulation rescued both Ser5 hyperphosphorylation and Rbp1 degradation triggered by TPL. Camptothecin (CPT), derived from the plant Camptotheca acuminata, specifically inhibits topoisomerase 1 (Top1). We first found that CPT induced antisense transcription at divergent CpG islands promoter. Interestingly, by immunofluorescence experiments, CPT was found to induce a burst of R loop structures (DNA/RNA hybrids) at nucleoli and mitochondria. We then decided to investigate the role of Top1 in R loop homeostasis through a short interfering RNA approach (RNAi). Using DNA/RNA immunoprecipitation techniques coupled to NGS I found that Top1 depletion induces an increase of R loops at a genome-wide level. We found that such increase occurs on the entire gene body. At a subset of loci R loops resulted particularly stressed after Top1 depletion: some of these genes showed the formation of new R loops structures, whereas other loci showed a reduction of R loops. Interestingly we found that new peaks usually appear at tandem or divergent genes in the entire gene body, while losses of R loop peaks seems to be a feature specific of 3’ end regions of convergent genes.

Assessment of the resilience of coral reefs to natural and human disturbances by means of recruitment panels in Indo-Pacific

Indo-Pacific region encompasses about 75% of world's coral reefs, but hard coral cover in this region experienced a 32% region-wide decline since 1970s. This great change is primarily ascribable to natural and anthropogenic pressures, including climate change and human activities effects. Coral reef conservation requires management strategies oriented to maintain their diversity and the capacity to provide ecosystem goods and services. Coral reef resilience, i.e. the capacity to recover after disturbances, is critical to their long-term persistence. The aims of the present study were to design and to test field experiments intended to measure changes in recruitment processes, as a fundamental aspect of the coral reef resilience. Recruitment experiments, using artificial panels suspended in the water column, were carried out in two Indo-Pacific locations affected by different disturbances: a new mine in Bangka Island (Indonesia), and the increased sedimentation due to coastal dynamics in Vavvaru Island (Maldives). One (or more) putatively disturbed site(s) was selected to be tested against 3 randomly selected control sites. Panels’ arrangement simulates 2 proximities to living corals, i.e. the sources of propagules: few centimetres and 2 meters over. Panels were deployed simultaneously at each site and left submerged for about five months. Recruits were identified to the lowest possible taxonomic level and recruited assemblages were analysed in terms of percent cover. In general it was not possible to detect significant differences between the benthic assemblages recruited in disturbed and control sites. The high variability observed in recruits assemblages structure among control sites may be so large to mask the possible disturbance effects. Only few taxa showed possible effects of the disturb they undergo. The field tests have highlighted strengths and weaknesses of the proposed approach and, based on these results, some possible improvements were suggested.

Planetary Gear Modal Vibration Experiments and Correlation against Lumped-Parameter and Finite Element Models

Experimental modal analysis techniques are applied to characterize the planar dynamic behavior of two spur planetary gears. Rotational and translational vibrations of the sun gear, carrier, and planet gears are measured. Experimentally obtained natural frequencies, mode shapes, and dynamic response are compared to the results from lumped-parameter and finite element models. Two qualitatively different classes of mode shapes in distinct frequency ranges are observed in the experiments and confirmed by the lumped-parameter model, which considers the accessory shafts and fixtures in the system to capture all of the natural frequencies and modes. The finite element model estimates the high-frequency modes that have significant tooth mesh deflection without considering the shafts and fixtures. The lumped-parameter and finite element models accurately predict the natural frequencies and modal properties established by experimentation. Rotational, translational, and planet mode types presented in published mathematical studies are confirmed experimentally. The number and types of modes in the low-frequency and high-frequency bands depend on the degrees of freedom in the central members and planet gears, respectively. The accuracy of natural frequency prediction is improved when the planet bearings have differing stiffnesses in the tangential and radial directions, consistent with the bearing load direction. (C) 2012 Elsevier Ltd. All rights reserved.

Enhancing Sensitivity In Natural Product Nmr: Exploring Optimal Non-Uniform Sampling

Recent optimizations of NMR spectroscopy have focused their attention on innovations in new hardware, such as novel probes and higher field strengths. Only recently has the potential to enhance the sensitivity of NMR through data acquisition strategies been investigated. This thesis has focused on the practice of enhancing the signal-to-noise ratio (SNR) of NMR using non-uniform sampling (NUS). After first establishing the concept and exact theory of compounding sensitivity enhancements in multiple non-uniformly sampled indirect dimensions, a new result was derived that NUS enhances both SNR and resolution at any given signal evolution time. In contrast, uniform sampling alternately optimizes SNR (t < 1.26T2) or resolution (t~3T2), each at the expense of the other. Experiments were designed and conducted on a plant natural product to explore this behavior of NUS in which the SNR and resolution continue to improve as acquisition time increases. Possible absolute sensitivity improvements of 1.5 and 1.9 are possible in each indirect dimension for matched and 2x biased exponentially decaying sampling densities, respectively, at an acquisition time of ¿T2. Recommendations for breaking into the linear regime of maximum entropy (MaxEnt) are proposed. Furthermore, examination into a novel sinusoidal sampling density resulted in improved line shapes in MaxEnt reconstructions of NUS data and comparable enhancement to a matched exponential sampling density. The Absolute Sample Sensitivity derived and demonstrated here for NUS holds great promise in expanding the adoption of non-uniform sampling.

Micrometer-sized ice particles for planetary-science experiments – II. Bidirectional reflectance

We have measured the bidirectional reflectance of spherical micrometer-sized water-ice particles in the visible spectral range over a wide range of incidence and emission angles. The small ice spheres were produced by spraying fine water droplets directly into liquid nitrogen. The resulting mean particle radii are 1.47 + 0.96 - 0.58 μm. Such a material shares many properties with ice in comets and at the surface of icy satellites. Measurements show that the fresh sample material is highly backscattering, contrasting with natural terrestrial snow and frost. The formation of agglomerates of particles during the sample production results in a noticeable variability of the photometric properties of the samples in their initial state. We have also observed significant temporal evolutions of the scattering behavior of the samples, shifting towards more forward scattering within some tens of hours, resulting most likely from sintering processes. All reflectance data are fitted by the Hapke photometric model (1993 and 2002 formulation) with a one/two/three-parameter Henyey-Greenstein phase function and the resulting Hapke parameters are provided. These parameters can be used to compare laboratory results with the observed photometric behaviors of astronomical objects. We show, in particular, that the optical properties of the fresh micrometer-sized ice samples can be used to reproduce the predominant backscattering in the phase curves of Enceladus and Europa.

Mechanism of surgical stress impairment of murine natural killer cell cytotoxicity

Natural killer cells may provide an important first line of defense against metastatic implantation of solid tumors. This antitumor function occurs during the intravascular and visceral lodgment phase of cancer dissemination, as demonstrated in small animal metastasis models. The role of the NK cell in controlling human tumor dissemination is more difficult to confirm, at least partially because of ethical restraints on experimental design. Nonetheless, a large number of solid tumor patient studies have demonstrated NK cell cytolysis of both autologous and allogeneic tumors.^ Of the major cancer therapeutic modalities, successful surgery in conjunction with other treatments offers the best possibility of cure. However, small animal experiments have demonstrated that surgical stress can lead to increased rates of primary tumor take, and increased incidence, size, and rapidity of metastasis development. Because the physiologic impact of surgical stress can also markedly impair perioperative antitumor immune function in humans, we examined the effect of surgical stress on perioperative NK cell cytolytic function in a murine preclinical model. Our studies demonstrated that hindlimb amputation led to a marked impairment of postoperative NK cell cytotoxicity. The mechanism underlying this process is complex and involves the postsurgical generation of splenic erythroblasts that successfully compete with NK cells for tumor target binding sites; NK cell-directed suppressor cell populations; and a direct impairment of NK cell recycling capacity. The observed postoperative NK cell suppression could be prevented by in vivo administration of pyrimidinone biologic response modifiers or by short term in vitro exposure of effector cells to recombinant Interleukin-2. It is hoped that insights gained from this research may help in the future development of NK cell specific perioperative immunotherapy relevant to the solid tumor patients undergoing cancer resection. ^

ANALYSIS OF THE CORYNEBACTERIUM PARVUM-INDUCED DECLINE OF MURINE NATURAL KILLER CELL-MEDIATED CYTOTOXICITY

Treatment of mice with the immunomodulating agent, Corynebacterium parvum (C. parvum), was shown to result in a severe and long-lasting depression of splenic natural killer (NK) cell-mediated cytotoxicity 5-21 days post-inoculation. Because NK cells have been implicated in immunosurveillance against malignancy (due to their spontaneous occurrence and rapid reactivity to a variety of histological types of tumors), as well as in resistance to established tumors, this decreased activity was of particular concern, since this effect is contrary to that which would be considered therapeutically desirable in cancer treatment (i.e. a potentiation of antitumor effector functions, including NK cell activity, would be expected to lead to a more effective destruction of malignant cells). Therefore, an analysis of the mechanism of this decline of splenic NK cell activity in C.parvum treated mice was undertaken.^ From in vitro co-culturing experiments, it was found that low NK-responsive C. parvum splenocytes were capable of reducing the normally high-reactivity of cells from untreated syngeneic mice to YAC-1 lymphoma, suggesting the presence of NK-directed suppressor cells in C. parvum treated animals. This was further supported by the demonstration of normal levels of cytotoxicity in C. parvum splenocyte preparations following Ficoll-Hypaque separation, which coincided with removal of the NK-suppressive capabilities of these cells. The T cell nature of these regulatory cells was indicated by (1) the failure of C. parvum to cause a reduction of NK cell activity, or the generation of NK-directed suppressor cells in T cell-deficient athymic mice, (2) the removal of C. parvum-induced suppression by T cell-depleting fractionation procedures or treatments, and (3) demonstration of suppression of NK cell activity by T cell-enriched C. parvum splenocytes. These studies suggest, therefore, that the eventual reduction of suppression by T cell elimination and/or inhibition, may result in a promotion of the antitumor effectiveness of C. parvum due to the contribution of "freed" NK effector cell activity.^ However, the temporary suppression of NK cell activity induced by C. parvum (reactivity of treated mice returns to normal levels within 28 days after C. parvum injection), may in fact be favorable in some situations, e.g. in bone marrow transplantation cases, since NK cells have been suggested to play a role also in the process of bone marrow graft rejection.^ Therefore, the discriminate use of agents such as C. parvum may allow for the controlled regulation of NK cell activity suggested to be necessary for the optimalization of therapeutic regimens. ^

Natural antisense transcripts of Trifolium repens dehydrins

The recently described complex nature of some dehydrin-coding sequences in Trifolium repens could explain the considerable variability among transcripts originating from a single gene.1 For some of the sequences the existence of natural antisense transcripts (NAT s), which could form sense-antisense (SAS) pairs, was predicted. The present study demonstrates that cis-natural antisense transcripts of 2 dehydrin types (YnKn and YnSKn) accumulate in white clover plants subjected to treatments with polyethylene glycol (PEG), abscisic acid (ABA), and high salt concentration. The isolated YnKn cis-NAT s mapped to sequence site enriched in alternative start codons. Some of the sense-antisense pairs exhibited inverse expression with differing profiles which depended on the applied stress. A natural antisense transcript coding for an ABC F family protein (a trans-NAT) which shares short sequence homology with YnSKn dehydrin was identified in plants subjected to salt stress. Forthcoming experiments will evaluate the impact of NAT s on transcript abundances, elucidating the role of transcriptional and post-transcriptional interferences in the regulation of dehydrin levels under various abiotic stresses.