939 resultados para Mitotic Exit


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This paper investigated the problem of confined flow under dams and water retaining structuresusing stochastic modelling. The approach advocated in the study combined a finite elementsmethod based on the equation governing the dynamics of incompressible fluid flow through aporous medium with a random field generator that generates random hydraulic conductivity basedon lognormal probability distribution. The resulting model was then used to analyse confined flowunder a hydraulic structure. Cases for a structure provided with cutoff wall and when the wall didnot exist were both tested. Various statistical parameters that reflected different degrees ofheterogeneity were examined and the changes in the mean seepage flow, the mean uplift forceand the mean exit gradient observed under the structure were analysed. Results reveal that underheterogeneous conditions, the reduction made by the sheetpile in the uplift force and exit hydraulicgradient may be underestimated when deterministic solutions are used.

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Introduction: Age-related macular degeneration (AMD) is a leading cause of vision loss in the elderly mostly due to the development of neovascular AMD (nAMD) or geographic atrophy (GA). Intravitreal injections of anti-vascular endothelial growth factor (VEGF) agents are an effective therapeutic option for nAMD. Following anti-VEGF treatments, increased atrophy of the retinal pigment epithelium (RPE) and choriocapillaries that resembles GA has been reported. We sought to evaluate the underlying genetic influences that may contribute to this process. Methods: We selected 68 single nucleotide polymorphisms (SNPs) from genes previously identified as susceptibility factors in AMD, along with 43 SNPs from genes encoding the VEGF protein and its cognate receptors as this pathway is targeted by treatment. We enrolled 467 consecutive patients (Feb 2009 to October 2011) with nAMD who received anti-VEGF therapy. The acutely presenting eye was designated as the study eye and retinal tomograms graded for macular atrophy at study exit. Statistical analysis was performed using PLINK to identify SNPs with a P value < 0.01. Logistic regression models with macular atrophy as dependent variable were fitted with age, gender, smoking status, common genetic risk factors and the identified SNPs as explanatory variables. Results: Grading for macular atrophy was available in 304 study eyes and 70% (214) were classified as showing macular atrophy. In the unadjusted analysis we observed significant associations between macular atrophy and two independent SNPs in the APCS gene: rs6695377: odds ratio (OR) = 1.98; 95% confidence intervals (CI): 1.23, 3.19; P = 0.004; rs1446965: OR = 2.49, CI: 1.29, 4.82; P = 0.006 and these associations remained significant after adjustment for covariates. Conclusions: VEGF is a mitogen and growth factor for choroidal blood vessels and the RPE and its inhibition could lead to atrophy of these key tissues. Anti-VEGF treatment can interfere with ocular vascular maintenance and may be associated with RPE and choroidal atrophy. As such, these medications, which block the effects of VEGF, may influence the development of GA. The top associated SNPs are found in the APCS gene, a highly conserved glycoprotein that encodes Serum amyloid P (SAP) which opsonizes apoptotic cells. SAP can bind to and activate complement components via binding to C1q, a mechanism by which SAP may remove cellular debris, affecting regulation of the three complement pathways.

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The aim of this study was to increase understanding of the mechanism and dominant drivers influencing phase separation during ram extrusion of calcium phosphate (CaP) paste for orthopaedic applications. The liquid content of extrudate was determined, and the flow of liquid and powder phases within the syringe barrel during extrusion were observed, subject to various extrusion parameters. Increasing the initial liquid-to-powder mass ratio, LPR, (0.4-0.45), plunger rate (5-20 mm/min), and tapering the barrel exit (45°-90°) significantly reduced the extent of phase separation. Phase separation values ranged from (6.22 ± 0.69 to 18.94 ± 0.69 %). However altering needle geometry had no significant effect on phase separation. From powder tracing and liquid content determination, static zones of powder and a non-uniform liquid distribution was observed within the barrel. Measurements of extrudate and paste LPR within the barrel indicated that extrudate LPR remained constant during extrusion, while LPR of paste within the barrel decreased steadily. These observations indicate the mechanism of phase separation was located within the syringe barrel. Therefore phase separation can be attributed to either; (1) the liquid being forced downstream by an increase in pore pressure as a result of powder consolidation due to the pressure exerted by the plunger or (2) the liquid being drawn from paste within the barrel, due to suction, driven by dilation of the solids matrix at the barrel exit. Differentiating between these two mechanisms is difficult; however results obtained suggest that suction is the dominant phase separation mechanism occurring during extrusion of CaP paste.

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The development and optimization of catalysts and catalytic processes requires knowledge of reaction kinetics and mechanisms. In traditional catalyst kinetic characterization, the gas composition is known at the inlet, and the exit flow is measured to determine changes in concentration. As such, the progression of the chemistry within the catalyst is not known. Technological advances in electromagnetic and physical probes have made visualizing the evolution of the chemistry within catalyst samples a reality, as part of a methodology commonly known as spatial resolution. Herein, we discuss and evaluate the development of spatially resolved techniques, including the evolutions and achievements of this growing area of catalytic research. The impact of such techniques is discussed in terms of the invasiveness of physical probes on catalytic systems, as well as how experimentally obtained spatial profiles can be used in conjunction with kinetic modelling. Furthermore, some aims and aspirations for further evolution of spatially resolved techniques are considered.

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Development of cribriform morphology (CM) heralds malignant change in human colon but lack of mechanistic understanding hampers preventive therapy. This study investigated CM pathobiology in three-dimensional (3D) Caco-2 culture models of colorectal glandular architecture, assessed translational relevance and tested effects of 1,25(OH)2D3, the active form of vitamin D. CM evolution was driven by oncogenic perturbation of the apical polarity (AP) complex comprising PTEN, CDC42 and PRKCZ (phosphatase and tensin homolog, cell division cycle 42 and protein kinase C zeta). Suppression of AP genes initiated a spatiotemporal cascade of mitotic spindle misorientation, apical membrane misalignment and aberrant epithelial configuration. Collectively, these events promoted “Swiss cheese-like” cribriform morphology (CM) comprising multiple abnormal “back to back” lumens surrounded by atypical stratified epithelium, in 3D colorectal gland models. Intestinal cancer driven purely by PTEN-deficiency in transgenic mice developed CM and in human CRC, CM associated with PTEN and PRKCZ readouts. Treatment of PTEN-deficient 3D cultures with 1,25(OH)2D3 upregulated PTEN, rapidly activated CDC42 and PRKCZ, corrected mitotic spindle alignment and suppressed CM development. Conversely, mutationally-activated KRAS blocked 1,25(OH)2D3 rescue of glandular architecture. We conclude that 1,25(OH)2D3 upregulates AP signalling to reverse CM in a KRAS wild type (wt), clinically predictive CRC model system. Vitamin D could be developed as therapy to suppress inception or progression of a subset of colorectal tumors.

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The photocatalytic reactor described in the NOx removal ISO 22197-1:2007 is used to study the kinetics of the process, using a film of P25 TiO2 which has either been conventionally pre-irradiated in a stream of air, or unconventionally in a stream of NO (1 ppmv). In the former case it is shown that the system does not achieve steady state exit levels of NO, probably due to the gradual accumulation of HNO3 on the surface of the photocatalyst. The NO-preconditioned TiO2 film demonstrated excellent steady-state levels when monitored as a function of NO concentration, [NO] and UV irradiance, ρ. However, in this case the photocatalytic reaction under study is NOT NOx removal, but the conversion of NO to NO2. It is shown that the kinetics of this steady state process fit very well to a kinetic expression based on a disrupted adsorption reaction mechanism, which has also been used by others to fit their observed (non-steady state) kinetics for NOx removal on conventionally-(air) preconditioned films of P25. The appropriateness of this model for either system is questioned, since in both systems the kinetics appear to have a significant mass transport element. These findings suggest that mass transport and non-steady-state kinetics are likely to be significant features for most active photocatalytic samples, where the %NO conversion is >7%, and so limits the usefulness of the NOx removal ISO 22197-1:2007.

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Chromosome structure and behaviour in both meiosis of the germ cells and mitosis of the embryo from fertilisation to the two-cell stage in Bursaphelenchus xylophilus were examined by DAPI staining and three-dimensional reconstruction of serial-section images from confocal laser-scanning microscopy. By this method, each chromosome’s shape and behaviour were clearly visible in early embryogenesis from fertilisation through the formation and fusion of the male and female pronuclei to the first mitotic division. The male pronucleus was bigger than that of the female, although the oocyte is larger and richer in nutrients than the sperm. From the shape of the separating chromosomes at anaphase, the mitotic chromosomes appeared to be polycentric or holocentric rather than monocentric. Each chromosome was clearly distinguishable in the male and female germ cells, pronuclei of the one-cell stage embryo, and the early embryonic nuclei. The haploid number of chromosomes (N) was six (2n = 12), and all chromosomes appeared similar. The chromosome pair containing the ribosomal RNA-coding site was visualised by fluorescence in situ hybridisation. Unlike the sex determination system in Caenorhabditis elegans (XX in hermaphrodite and XO in male), the system for B. xylophilus may consist of an XX female and an XY male.

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As proteínas existentes nas células são produzidas pelo mecanismo de tradução do mRNA, no qual a informação genética contida nos genes é descodificada em cadeias polipeptídicas. O código genético, que define as regras de descodificação do genoma, minimiza os erros de tradução do mRNA, garantindo a síntese de proteínas com elevada fidelidade. Esta é essencial para a estabilidade do proteoma e para a manutenção e funcionamento dos processos celulares. Em condições fisiológicas normais, os erros da tradução do mRNA ocorrem com frequências que variam de 10-3 a 10-5 erros por codão descodificado. Situações que aumentam este erro basal geralmente estão associadas ao envelhecimento, stresse e a doenças; no entanto, em certos organismos o código genético é traduzido naturalmente com elevado erro, indicando que a síntese de proteínas aberrantes pode de algum modo ser vantajosa. A fim de estudar a resposta celular aos erros de tradução do mRNA, construímos leveduras que incorporam serina no proteoma em resposta a um codão de leucina, usando a expressão constitutiva de um tRNASer mutante. Este fenómeno genético artificial provocou uma forte diminuição da esporulação, da viabilidade e da eficiência de mating, afectando imensamente a reprodução sexual da levedura. Observou-se também uma grande heterogeneidade no tamanho e na forma das células e elevada instabilidade genómica, com o aparecimento de populações poliplóides e aneuplóides. No sentido de clarificar as bases celulares e moleculares daqueles fenótipos e compreender melhor a biologia do erro de tradução do mRNA, construímos também células de levedura que inserem serina em resposta a um codão de leucina de modo indutível e controlado. Utilizaram-se perfis de mRNA total e de mRNA associado a polissomas para elucidar a resposta celular ao erro de tradução do mRNA. Observou-se a indução de genes envolvidos na resposta ao stresse geral, stresse oxidativo e na unfolded protein response (UPR). Um aumento significativo de espécies reactivas de oxigénio (ROS) e um forte impacto negativo na capacidade das células pós-mitóticas re-iniciarem o crescimento foram também observados. Este fenótipo de perda de viabilidade celular foi resgatado por scavangers de ROS, indicando que o stresse oxidativo é a principal causa de morte celular causada pelos erros de tradução. Este estudo levanta a hipótese de que o stresse oxidativo e a acumulação de ROS, ao invés do colapso súbito do proteoma, são as principais causas da degeneração celular e das doenças humanas associadas aos erros de tradução do genoma. ABSTRACT: Proteins are synthesized through the mechanism of translation, which uses the genetic code to transform the nucleic acids based information of the genome into the amino acids based information of the proteome. The genetic code evolved in such a manner that translational errors are kept to a minimum and even when they occur their impact is minimized by similar chemical properties of the amino acids. Protein synthesis fidelity is essential for proteome stability and for functional maintenance of cellular processes. Indeed, under normal physiological conditions, mistranslation occurs at frequencies that range from 10-3 to 10-5 errors per codon decoded. Situations where this basal error frequency increases are usually associated to aging and disease. However, there are some organisms where genetic code errors occur naturally at high level, suggesting that mRNA mistranslation can somehow be beneficial. In order to study the cellular response to mRNA mistranslation, we have engineered single codon mistranslation in yeast cells, using constitutive expression of mutant tRNASer genes. These mistranslating strains inserted serines at leucine-CUG sites on a proteome wide scale due to competition between the wild type tRNALeu with the mutant tRNASer. Such mistranslation event decreased yeast sporulation, viability and mating efficiencies sharply and affected sexual reproduction strongly. High heterogeneity in cell size and shape and high instability in the genome were also observed, with the appearance of some polyploid or aneuploid cell populations. To further study the cellular and molecular basis of those phenotypes and the biology of mRNA mistranslation, we have also engineered inducible mRNA misreading in yeast and used total mRNA and polysome associated mRNA profiling to determine whether codon misreading affects gene expression. Induced mistranslation up-regulated genes involved in the general stress response, oxidative stress and in the unfolded protein response (UPR). A significant increase in reactive oxygen species (ROS) and a strong negative impact on the capacity of post-mitotic cells to re-initiate growth in fresh media were also observed. This cell viability phenotype was rescued by scavengers of ROS, indicating that oxidative stress is the main cause of cell death caused by mRNA mistranslation. This study provides strong support for the hypothesis that oxidative stress and ROS accumulation, rather than sudden proteome collapse or major proteome disruption, are the main cause of the cellular degeneration observed in human diseases associated mRNA mistranslation.

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Os dinoflagelados são um grupo muito diverso de protistas que possuem um conjunto de características pouco comuns. Os peridinióides são dinoflagelados com teca que é formada por seis séries latitudinais de placas, incluindo a série cingular e um anel incompleto de placas intercalares anteriores, embora as últimas estejam ausentes em algumas espécies de Peridiniopsis. São dinoflagelados com simetria bilateral em relação ao plano apical que contem o eixo dorso-ventral. Na série sulcal há apenas uma placa posterior que contacta com o limite ventral de duas grandes placas antapicais. Entre os peridinióides, a presença ou ausência de um poro apical e o número de placas no cíngulo são geralmente consideradas marcas filogenéticas importantes ao nível de género ou família. Actualmente, a definição de Peridinium Ehrenberg, o dinoflagelado mais comum de água doce, inclui organismos com combinações diferentes destas duas características. Trabalhos anteriores sobre a ultrastrutura e afinidade filogenética das espécies tipo de Peridinium, P. cinctum, e Peridiniopsis Lemmermann, P. borgei também sugerem a necessidade de reexaminar as relações taxonómicas dos peridinióides. Esta tese combina o estudo ultrastrutural de uma selecção de espécies com hipóteses filogenéticas baseadas nas sequências de LSU rDNA, para aumentar o nosso conhecimento das diferenças e afinidades dentro dos peridinióides. Tem como objectivo aumentar o nosso conhecimento das características individuais das células que possam levar a reconhecer sinapomorfias que possam ser usadas como marcadores dos peridinióides como um todo e dos seus subgrupos. As espécies escolhidas para exame pormenorizado foram: Peridinium palatinum Lauterborn, de um grupo com duas placas intercalares anteriores, seis placas cingulares e sem poro apical; Peridinium lomnickii Wo!oszy"ska, de um grupo com poro apical, três placas intercalares e seis cingulares; Peridiniopsis berolinensis (Lemmermann) Bourrelly, uma espécie heterotrófica com poro apical, sem placas intercalares e com seis placas cingulares; e Sphaerodinium cracoviense Wo!oszy"ska, um membro de um género de formas com teca com um tipo de tabulação marginalmente peridinióide, com um suposto poro apical e quatro placas intercalares anteriores. Peridinium palatinum difere de Peridinium e Peridiniopsis típicos, quer em características da teca, quer internas. As diferenças estimadas entre as sequências parciais de LSU rDNA de P. palatinum e a espécie próxima P. pseudolaeve, relativamente a P. cinctum são comparativamente grandes e, juntamente com a topologia da árvore filogenética, apoiam a separação de P. palatinum e formas próximas ao nível de género. Palatinus nov. gen. foi, então, descrito com as novas combinações Palatinus apiculatus nov. comb. (espécie tipo; sin. Peridinium palatinum), P. apiculatus var. laevis nov. comb. e P. pseudolaevis nov. comb.. As características distintivas de Palatinus incluem uma superfície das placas lisa ou um tanto granulosa, mas não areolada, um grande pirenóide central penetrado por canais citoplasmáticos e de onde radiam lobos plastidiais, e a presença de uma fiada microtubular homóloga à de um pedúnculo. As células de Palatinus saem da teca pela zona antapicalpos- cingular. Peridinium lomnickii apresenta tabulação semelhante às formas marinhas, produtoras de quistos calcários, do género Scrippsiella A.R. Loeblich. Para comparação, adicionámos novas observações ultrastruturais de S. trochoidea. Peridinium lomnickii tem uma combinação de características diferente de Peridinium, Peridiniopsis e Scrippsiella. As hipóteses filogenéticas baseadas em DNA colocam P. lomnickii no mesmo ramo que Pfiesteria Steidinger et Burkholder, Tyrannodinium e outras Pfiesteriaceae, com as quais partilha um "microtubular basket" e uma ligação peculiar entre duas placas do sulco. As características distintivas do novo género proposto Chimonodinium gen. ined. incluem, além da tabulação, a ausência de pirenóides, a presença de um "microtubular basket" com quatro ou cinco fiadas sobrepostas de microtúbulos associados a um pequeno pedúnculo, um sistema pusular com tubos pusulares bem definidos ligados aos canais flagelares, e a produção de quistos não calcários. Peridiniopsis berolinensis partilha várias características significativas com Pfiesteria e afins, como um "microtubular basket" com a capacidade de suportar um tubo de alimentação, quimiossensibilidade para encontrar presas apropriadas, o modo de natação junto às presas e a organização geral da célula. Hipóteses filogenéticas com base em LSU rDNA confirmam a afinidade entre P. berolinensis e Pfiesteria bem como a relação mais remota com a espécie tipo de Peridiniopsis, P. borgei. Estas razões justificam a proposta de Tyrannodinium gen. nov., uma nova Pfiesteriaceae que difere de outros membros do grupo por viver em água doce e nos pormenores da tabulação. Sphaerodinium cracoviense revelou a tabulação típica do género Sphaerodinium, que apresenta um número de placas intercalares superiores e pos-cingulares maior que o que é típico em peridinióides: 4 e 6, respectivamente. Observações em SEM mostraram uma estrutura apical diferente da dos peridinióides, e um sulco apical numa das placas fazendo lembrar a área apical de alguns woloszynskióides. Os pormenores do aparelho flagelar e do sistema pusular ligam o Sphaerodinium aos woloszynskióides em geral e ao género Baldinia em particular, mas não aos peridinióides. O volumoso estigma de S. cracoviense revelou ser extraplastidial e de um modelo único, composto por elementos que se encontram em woloszynskióides, mas nunca encontrados anteriormente juntos. A análise filogenética baseada nas sequências parciais de LSU rDNA também sugerem uma maior proximidade de S. cracoviense com os woloszynskióides do que com os peridinióides. Futuras análises pormenorizadas de dinoflagelados peridinióides, em especial entre os do numeroso grupo de espécies com poro apical, serão necessárias para clarificar as suas relações taxonómicas; e a produção de descrições melhoradas das características finas particulares das células serão um requisito para perceber a evolução dos caracteres dos peridinióides por forma a podermos identificar marcadores filogenéticos.

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Objectives. Mindfulness meditation practices have become increasingly popular in clinical therapies, changing patterns of depressogenic thinking for individuals who experience consecutive episodes of depression. We were interested in finding out how Mindfulness Based Cognitive Therapy (MBCT) worked for programme participants by focussing on how meditative practices changed their relationships to their thoughts. Design. Data for the study came from six semi-structured research interviews carried out with individuals who had taken part in an 8 week MBCT programme Methods. We used Interpretative Phenomenological Analysis (IPA) to analyse the experiential accounts. Results. We report on two superordinate themes – Engaging the Neutral Mind (with subordinate themes ‘breaking the paralysis of worry’ and ‘choosing to think differently’) and Experiencing the Neutral Mind (with subordinate themes of ‘reflection on previous thinking styles’ and ‘becoming psychologically self-reliant’). Conclusions. Themes from the present study offer support to the assertion that mindfulness meditation helps facilitate a different mode of meta-cognitive processing with which to handle depression-related cognitions. Practitioner Points Participants reported that they experienced an enhanced capacity to differentiate between their thought processes, experiencing an ability to tolerate some more uncomfortable thoughts and experiencing a/more choice in how to respond to thoughts Participants recognised that ruminating over negative thoughts was related to depressive states and experienced a shift in meta-cognitive processes that actively challenged depressogenic cognitions Participants became more psychologically self-reliant and therapeutically independent following MBCT Integrating mindfulness based practices in therapy may be a mediating factor in sustaining psychological wellbeing and may help clients develop self-compassion Future research looks to examining exit cases to understand elements of MBCT which are experienced as less successful by clients

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Since its foundation, militant democratic arguments have underpinned an enforced secularism in Turkey. The 2002 election of the AKP, described as a “moderate Islamist party”, has challenged Turkey’s secular identity. In the more than twelve years since the AKP has been in power, Turkey’s political landscape has experienced significant changes, with periods of extensive democratic reforms punctuated by regression in certain areas, notably freedom of expression and the right to protest. State repressive measures coupled with Recep Tayyip Erdoğan’s reluctance to exit the political stage have been the focus of much commentary and analysis. This article argues, however, that under AKP rule the Kurdish issue – critical to ensuring the normalization of politics and democratization in Turkey – has been brought in from the political cold and assesses the creation and role of the HDP (Halkların Demokratik Partisi), a Kurdish political party that is endeavoring to situate itself in the mainstream of Turkey’s political landscape. We posit that the HDP can be viewed as the offspring of this “democratic opening,” a project that was meant to ensure a radical transformation of the Kurdish issue in Turkey. Through analysing the historical trajectory of both AKP and HDP and the militant democratic arguments that led to their predecessors’ exclusion from the public sphere, this article engages with the key question of the extent to which the AKP’s treatment of the Kurdish issue has provided a vehicle for broader democratisation and facilitated a reconsideration of the Kurdish question in Turkey.

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Tese de dout., Química, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2008

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Understanding heart development on a molecular level is a requirement for uncovering the causes of congenital heart diseases. Several genes have been implicated as critical for heart development. However, the inducers of these genes as well as their targets and pathways, remain largely unknown. We have identified a promoter element of chick cCer able to drive EGFP expression in a population of cells that consistently exit from the anterior primitive streak region, from as early as stage HH3+, and that later will populate the heart. Using this promoter element as a tool allowed us to identify novel genes previously not known to potentially play a role in heart development. In order to identify and study genes expressed and involved in the correct development and differentiation of the vertebrate heart precursor cell (HPC) lineages, a differential screening using Affymetrix GeneChip® system technologies was performed. Remarkably, this screening led to the identification of more than 700 transcripts differentially expressed in the heart forming regions (HFR). Bioinformatic tools allowed us to filter the large amount of data generated from this approach and to select a few transcripts for in vivo validation. Five genes were selected for further characterization by whole mount in situ hybridization leading to the validation of their expression in the HPC. From those, Adtk1 and Ccbe1 were selected for functional analysis. Regarding to ccbe1, a more detailed WISH analysis was performed and showed that Ccbe1 is expressed specifically on the cardiac progenitors regions at HH4, more specifically in primary heart field and at later stages is present in the second heart field. Further functional analyses by knockdown and overexpression revealed an important role for Ccbe1 in early heart tube formation. Moreover, the results presented in this thesis suggested that Ccbe1 is a key gene during heart development and might be limited to multipotent and highly proliferative progenitors and downregulated upon cellular commitment into more specific cardiac phenotypes. Other of the genes identified, Adtk1 was also subjected to further functional studies. Knockdown of Adtk1 using morpholino oligonucleotides suggested that it might be necessary for the migration and fusion of the heart tube as well as for neural tube closure.

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Dissertação de mestrado, Engenharia Informática, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

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Dissertação de mestrado, Biologia Marinha, Faculdade de Ciências e Tecnologia, Universidade do Algarve; Instituto Español de Oceanografia; 2015