945 resultados para SOD MIMICS


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In aerobic organisms, protection against oxidative damage involves the combined action of highly specialized antioxidant enzymes, such as superoxide dismutase (SOD) and catalase. Here we describe the isolation and characterization of another gene in the yeast Saccharomyces cerevisiae that plays a critical role in detoxification of reactive oxygen species. This gene, named ATX1, was originally isolated by its ability to suppress oxygen toxicity in yeast lacking SOD. ATX1 encodes a 8.2-kDa polypeptide exhibiting significant similarity and identity to various bacterial metal transporters. Potential ATX1 homologues were also identified in multicellular eukaryotes, including the plants Arabidopsis thaliana and Oryza sativa and the nematode Caenorhabditis elegans. In yeast cells, ATX1 evidently acts in the transport and/or partitioning of copper, and this role in copper homeostasis appears to be directly relevant to the ATX1 suppression of oxygen toxicity: ATX1 was incapable of compensating for SOD when cells were depleted of exogenous copper. Strains containing a deletion in the chromosomal ATX1 locus were generated. Loss of ATX1 function rendered both mutant and wild-type SOD strains hypersensitive toward paraquat (a generator of superoxide anion) and was also associated with an increased sensitivity toward hydrogen peroxide. Hence, ATX1 protects cells against the toxicity of both superoxide anion and hydrogen peroxide.

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Sequence analysis of the variable regions of the heavy and light chains of the anti-idiotypic antibody 6F9, which mimics the meningococcal group C capsular polysaccharide (MCP), was performed. The immunogenic site on 6F9 responsible for inducing an anti-MCP antibody response was determined by means of sequence and computer model analysis of these data. Complementarity-determining region 3 (CDR3) was found to be unique in that the sequence tract YRY was exposed on the surface. A synthetic peptide spanning the CDR3 domain was synthesized and complexed to proteosomes (meningococcal group B outer membrane protein). Immunizations of BALB/c mice with the peptide-proteosome complex resulted in a significant anti-MCP antibody response. Immunized mice were protected against infection with a lethal dose of Neisseria meningitidis serogroup C.

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Lagartos teiú eclodem no verão e enfrentam o desafio de crescer e armazenar substratos em um curto período de tempo, antes do início do período de jejum e depressão metabólica (≈80%) a temperaturas amenas durante o inverno (≈17 °C). No despertar, o aumento do metabolismo e a reperfusão de órgãos favoreceriam a ocorrência de estresse oxidativo. Na primeira parte do presente estudo investigou−se os ajustes que compatibilizam as demandas em teiús neonatos, especialmente na pré-hibernação, por meio da gravação do comportamento em vídeo e da análise da massa dos corpos gordurosos abdominais e do nível plasmático de corticosterona (CORT) durante o primeiro ciclo anual. No início do outono a massa corpórea dos teiús foi 27 g e o comprimento rostro−cloacal 9,3 cm e aumentaram 40% e 20%, respectivamente, ao longo do outono, enquanto que as taxas diminuíram progressivamente até atingirem o valor zero no início do inverno. Na primavera, a massa corpórea dos teiús aumentou 80% em relação ao despertar e dobrou em relação ao final do verão; o comprimento acumulou um aumento de 27% em relação ao final do verão. A massa relativa dos corpos gordurosos foi 3,7% no início do outono e diminuiu nos meses subsequentes; no despertar, este estoque acumulou uma perda de 63% da sua massa. No início do outono 74% dos teiús estavam ativos por 4,7 h e permaneceram 2 h assoalhando diariamente; ao longo do outono o número de animais ativos e o tempo em atividade diminuíram até que todos se tornaram inativos. Na primavera 83% dos teiús estavam ativos por 7 h e permaneceram 4 h assoalhando. Um padrão sazonal similar foi observado na atividade locomotora e na alimentação. No outono, a alimentação cessou antes da atividade diária e os teiús tornaram−se afágicos algumas semanas antes da entrada em hibernação. Os maiores níveis de CORT foram observados no início do outono, reduzindo progressivamente até valores 75 e 86% menores na dormência e despertar, respectivamente; na primavera os níveis de CORT foram 32% menores em comparação com o início do outono. Este padrão sugere um papel da CORT nos ajustes que promovem a ingestão de alimento e a deposição de substratos energéticos no outono. A redução da atividade geral no final do outono contribuiria para a economia energética e manutenção da massa corpórea, apesar da redução da ingestão de alimento. O curso temporal das alterações fisiológicas e comportamentais em neonatos reforça a ideia de que a dormência sazonal nos teiús é o resultado da expressão de um ritmo endógeno. Na segunda parte do estudo foi investigada a hipótese de que ocorreriam ajustes das defesas antioxidantes durante a hibernação, em antecipação ao despertar. Foram analisados marcadores de estresse oxidativo e antioxidantes em vários órgãos de teiús em diferentes fases do primeiro ciclo anual. A CS, um indicador do potencial oxidante, não variou no fígado e foi menor no rim e no pulmão na hibernação. As enzimas antioxidantes revelaram (1) um efeito abrangente de redução das taxas na hibernação e despertar; por exemplo, GR e CAT foram menores em todos órgãos analisados e a GST tendeu a diminuir no fígado e no rim, embora constante no coração e no pulmão. A G6PDH no fígado e no rim não variou. (2) No fígado, a GST, a Se−GPX e o teor de TBARS foram maiores na atividade de outono em relação à primavera e a Se−GPX permaneceu elevada na hibernação. (3) No fígado, a SOD foi maior na hibernação e despertar em relação ao outono e a Mn−SOD seguiu este padrão. Em contraste, no rim, coração e pulmão a SOD foi menor na hibernação e as taxas se recuperaram no coração e pulmão no despertar. A Mn−SOD seguiu este padrão no pulmão. A concentração e o estado redox da glutationa não variaram no fígado, rim e coração; no pulmão o teor de Eq−GSH e GSH foi menor na hibernação, com tendência à recuperação no despertar. O teor de PC no rim foi maior na hibernação e diminuiu no despertar. No fígado, as alterações no jejum se assemelham às sazonais, como sugerem a inibição da CAT e GR e aumento da Se−GPX. Os efeitos do jejum na primavera no rim diferem dos efeitos sazonais, como sugerem a redução do teor de Eq−GSH e GSH e o aumento da razão GSSG:GSH, a redução da G6PDH e o aumento de PC. No conjunto, houve um efeito predominante de redução das taxas enzimáticas na hibernação e no despertar, exceto pelas taxas aumentadas da SOD e Se−GPX no fígado e pela recuperação da SOD no coração e da GR, SOD e Mn−SOD no pulmão no despertar. As elevadas taxas das enzimas antioxidantes no teiú em comparação a outros ectotermos e a ausência de evidências de estresse oxidativo no despertar sugerem que a atividade enzimática remanescente é suficiente para prevenir danos aos tecidos face às flutuações do metabolismo

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O câncer de mama é o segundo tipo de neoplasia mais prevalente no mundo e o mais comum entre as mulheres. É descrito que o padrão de consumo alimentar materno e paterno está relacionado à suscetibilidade da prole ao desenvolvimento de doenças crônicas não transmissíveis, inclusive o câncer. A amora-preta é uma das frutas com maior conteúdo antioxidante e seus compostos bioativos possuem atividade antioxidante, anticarcinogênica e anti-inflamatória. Sendo assim, o presente trabalho propõe avaliar os efeitos do consumo materno e/ou paterno de extrato de amora-preta (Rubus spp.) na suscetibilidade da prole feminina ao desenvolvimento de neoplasias mamárias quimicamente induzidas. Para tanto, camundongos da linhagem C57BL/6 foram divididos aleatoriamente em 4 grupos: pai amora (PA), mãe amora (MA), pai e mãe amora (PMA) e controle (CTRL). Os pais receberam extrato de amora-preta logo após o desmame durante 8 semanas e as mães receberam o extrato durante a gestação e lactação. O extrato de amora-preta foi administrado na água de beber (0.84g de antocianinas/L) ad libitum. Os pais tratados com extrato de amora apresentaram redução na atividade enzimática da superóxido dismutase (SOD) e da catalase (CAT) no testículo (p<0.05 e p<0.001, respectivamente), aumento na capacidade antioxidante plasmática, na porcentagem de espermatozoides normais e na produção diária de espermatozóides em relação ao grupo controle (p<0.001 para todos). Além disso, os grupos PA, MA e PMA apresentaram aumento na taxa de prenhez (p<0.05) e redução da mortalidade perinatal (p<0.01, p<0.05 e p<0.001, respectivamente). Em relação à prole feminina não submetida à carcinogênese foi observada redução na capacidade antioxidante plasmática nos grupos PA (p<0.001) e MA (p<0.01), enquanto o grupo PMA apresentou aumento nesse parâmetro (p<0.001). No desenvolvimento da glândula mamária, houve aumento do desenvolvimento epitelial nos grupos PA, MA e PMA (p<0.001 para todos), de diferenciação nos grupos MA e PMA (p<0.01 para ambos) e da taxa de apoptose nos grupos MA e PMA (p<0.05), além de redução no número de TEBs nos grupos PA, MA e PMA (p<0.01, p<0.001 e p<0.001, respectivamente). Não foram observadas alterações significativas nas filhas submetidas à indução química da carcinogênese mamária por DMBA. Assim, é possível concluir que apesar de ter alterado o desenvolvimento da glândula mamária, o consumo materno e/ou paterno de extrato de amora-preta não foi capaz de impactar sobre a suscetibilidade da prole feminina à carcinogênese mamária quimicamente induzida.

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Este trabalho avalia a influência das emoções humanas expressas pela mímica da face na tomada de decisão de sistemas computacionais, com o objetivo de melhorar a experiência do usuário. Para isso, foram desenvolvidos três módulos: o primeiro trata-se de um sistema de computação assistiva - uma prancha de comunicação alternativa e ampliada em versão digital. O segundo módulo, aqui denominado Módulo Afetivo, trata-se de um sistema de computação afetiva que, por meio de Visão Computacional, capta a mímica da face do usuário e classifica seu estado emocional. Este segundo módulo foi implementado em duas etapas, as duas inspiradas no Sistema de Codificação de Ações Faciais (FACS), que identifica expressões faciais com base no sistema cognitivo humano. Na primeira etapa, o Módulo Afetivo realiza a inferência dos estados emocionais básicos: felicidade, surpresa, raiva, medo, tristeza, aversão e, ainda, o estado neutro. Segundo a maioria dos pesquisadores da área, as emoções básicas são inatas e universais, o que torna o módulo afetivo generalizável a qualquer população. Os testes realizados com o modelo proposto apresentaram resultados 10,9% acima dos resultados que usam metodologias semelhantes. Também foram realizadas análises de emoções espontâneas, e os resultados computacionais aproximam-se da taxa de acerto dos seres humanos. Na segunda etapa do desenvolvimento do Módulo Afetivo, o objetivo foi identificar expressões faciais que refletem a insatisfação ou a dificuldade de uma pessoa durante o uso de sistemas computacionais. Assim, o primeiro modelo do Módulo Afetivo foi ajustado para este fim. Por fim, foi desenvolvido um Módulo de Tomada de Decisão que recebe informações do Módulo Afetivo e faz intervenções no Sistema Computacional. Parâmetros como tamanho do ícone, arraste convertido em clique e velocidade de varredura são alterados em tempo real pelo Módulo de Tomada de Decisão no sistema computacional assistivo, de acordo com as informações geradas pelo Módulo Afetivo. Como o Módulo Afetivo não possui uma etapa de treinamento para inferência do estado emocional, foi proposto um algoritmo de face neutra para resolver o problema da inicialização com faces contendo emoções. Também foi proposto, neste trabalho, a divisão dos sinais faciais rápidos entre sinais de linha base (tique e outros ruídos na movimentação da face que não se tratam de sinais emocionais) e sinais emocionais. Os resultados dos Estudos de Caso realizados com os alunos da APAE de Presidente Prudente demonstraram que é possível melhorar a experiência do usuário, configurando um sistema computacional com informações emocionais expressas pela mímica da face.

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Differential SAR Interferometry (DInSAR) is a remote sensing method with the well demonstrated ability to monitor geological hazards like earthquakes, landslides and subsidence. Among all these hazards, subsidence involves the settlement of the ground surface affecting wide areas. Frequently, subsidence is induced by overexploitation of aquifers and constitutes a common problem that affects developed societies. The excessive pumping of underground water decreases the piezometric level in the subsoil and, as a consequence, increases the effective stresses with depth causing a consolidation of the soil column. This consolidation originates a settlement of ground surface that must be withstood by civil structures built on these areas. In this paper we make use of an advanced DInSAR approach - the Coherent Pixels Technique (CPT) [1] - to monitor subsidence induced by aquifer overexploitation in the Vega Media of the Segura River (SE Spain) from 1993 to the present. 28 ERS-1/2 scenes covering a time interval of about 10 years were used to study this phenomenon. The deformation map retrieved with CPT technique shows settlements of up to 80 mm at some points of the studied zone. These values agree with data obtained by means of borehole extensometers, but not with the distribution of damaged buildings, well points and basements, because the occurrence of damages also depends on the structural quality of the buildings and their foundations. The most interesting relationship observed is the one existing between piezometric changes, settlement evolution and local geology. Three main patterns of ground surface and piezometric level behaviour have been distinguished for the study zone during this period: 1) areas where deformation occurs while ground conditions remain altered (recent deformable sediments), 2) areas with no deformation (old and non-deformable materials), and 3) areas where ground deformation mimics piezometric level changes (expansive soils). The temporal relationship between deformation patterns and soil characteristics has been analysed in this work, showing a delay between them. Moreover, this technique has allowed the measurement of ground subsidence for a period (1993-1995) where no instrument information was available.

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Lidocaine bears in its structure both an aromatic ring and a terminal amine, which can be protonated at physiological pH, linked by an amide group. Since lidocaine causes multiple inhibitory actions on nicotinic acetylcholine receptors (nAChRs), this work was aimed to determine the inhibitory effects of diethylamine (DEA), a small molecule resembling the hydrophilic moiety of lidocaine, on Torpedo marmorata nAChRs microtransplanted to Xenopus oocytes. Similarly to lidocaine, DEA reversibly blocked acetylcholine-elicited currents (IACh) in a dose-dependent manner (IC50 close to 70 μM), but unlike lidocaine, DEA did not affect IACh desensitization. IACh inhibition by DEA was more pronounced at negative potentials, suggesting an open-channel blockade of nAChRs, although roughly 30% inhibition persisted at positive potentials, indicating additional binding sites outside the pore. DEA block of nAChRs in the resting state (closed channel) was confirmed by the enhanced IACh inhibition when pre-applying DEA before its co-application with ACh, as compared with solely DEA and ACh co-application. Virtual docking assays provide a plausible explanation to the experimental observations in terms of the involvement of different sets of drug binding sites. So, at the nAChR transmembrane (TM) domain, DEA and lidocaine shared binding sites within the channel pore, giving support to their open-channel blockade; besides, lidocaine, but not DEA, interacted with residues at cavities among the M1, M2, M3, and M4 segments of each subunit and also at intersubunit crevices. At the extracellular (EC) domain, DEA and lidocaine binding sites were broadly distributed, which aids to explain the closed channel blockade observed. Interestingly, some DEA clusters were located at the α-γ interphase of the EC domain, in a cavity near the orthosteric binding site pocket; by contrast, lidocaine contacted with all α-subunit loops conforming the ACh binding site, both in α-γ and α-δ and interphases, likely because of its larger size. Together, these results indicate that DEA mimics some, but not all, inhibitory actions of lidocaine on nAChRs and that even this small polar molecule acts by different mechanisms on this receptor. The presented results contribute to a better understanding of the structural determinants of nAChR modulation.

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MicroRNAs (miRNAs) control gene expression mostly post-transcriptionally by guiding transcript cleavage and/or translational repression of complementary mRNA targets, thereby regulating developmental processes and stress responses. Despite the remarkable expansion of the field, the mechanisms underlying miRNA activity are not fully understood. In this article, we describe a transient expression system in Arabidopsis mesophyll protoplasts, which is highly amenable for the dissection of miRNA pathways. We show that by transiently overexpressing primary miRNAs and target mimics, we can manipulate miRNA levels and consequently impact on their targets. Furthermore, we developed a set of luciferase-based sensors for quantifying miRNA activity that respond specifically to both endogenous and overexpressed miRNAs and target mimics. We demonstrate that these miRNA sensors can be used to test the impact of putative components of the miRNA pathway on miRNA activity, as well as the impact of specific mutations, by either overexpression or the use of protoplasts from the corresponding mutants. We further show that our miRNA sensors can be used for investigating the effect of chemicals on miRNA activity. Our cell-based transient expression system is fast and easy to set up, and generates quantitative results, being a powerful tool for assaying miRNA activity in vivo.

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Sediment porewater oxygen profiles were measured with micro and needle electrodes in sediment cores of 27 stations in the Skagerrak (northeastern North Sea). Oxygen penetration depth ranged from 3 to 20 mm depth. Fluxes estimated from the oxygen gradients varied from 3 to 18 mmol m**-2 d**-1. Oxygen penetration and flux depend on water depth, but possibly more on the hydrological conditions, related to the import of fresh organic matter by primary production in the water column. Oxygen fluxes were not related to the total organic carbon (TOC) content of the sediments. Stations in the eastern part of the Skagerrak showed high burial rates of TOC. At 6 stations porewater chemistry of Fe, Mn and NO3- was strongly associated with the oxygen distribution. The average relative contribution of terminal electron acceptors to carbon mineralisation was estimated at 85% for O2, 0.5% for Mn, 4.5% for [NO3]3-, 1% for Fe and 9% for [SO4]2-. At one station the occurrence of exceptionally high solid manganese oxyhydroxides was probably related to an active internal manganese cycle.

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Although ocean acidification is expected to impact (bio)calcification by decreasing the seawater carbonate ion concentration, [CO3]2-, there exists evidence of non-uniform response of marine calcifying plankton to low seawater [CO3]2-. This raises questions on the role of environmental factors other than acidification and on the complex physiological responses behind calcification. Here we investigate the synergistic effect of multiple environmental parameters, including temperature, nutrient (nitrate and phosphate) availability, and seawater carbonate chemistry on the coccolith calcite mass of the cosmopolitan coccolithophore Emiliania huxleyi, the most abundant species in the world ocean. We use a suite of surface (late Holocene) sediment samples from the South Atlantic and southwestern Indian Ocean taken from depths lying well above the modern lysocline. The coccolith calcite mass in our results presents a latitudinal distribution pattern that mimics the main oceanographic features, thereby pointing to the potential importance of phosphorus and temperature in determining coccolith mass by affecting primary calcification and possibly driving the E. huxleyi morphotype distribution. This evidence does not necessarily argue against the potentially important role of the rapidly changing seawater carbonate chemistry in the future, when unabated fossil fuel burning will likely perturb ocean chemistry beyond a critical point. Rather our study highlights the importance of evaluating the combined effect of several environmental stressors on calcifying organisms to project their physiological response(s) in a high CO2 world and improve interpretation of paleorecords.

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The modern eastern equatorial Pacific (EEP) is a major natural source for atmospheric carbon dioxide and is thought to be connected to high-latitude ocean dynamics by oceanic teleconnections on glacial-interglacial timescales. A wealth of sedimentary records aiming at reconstructing last Quaternary changes in primary productivity and nutrient utilization have been devoted to understanding those linkages between the EEP and other distant oceanic areas. Most of these records are, however, clustered in the pelagic EEP cold tongue, with comparatively little attention devoted to coastal areas. Here we present downcore measurements of the composition and concentration of the diatom assemblage together with opal (biogenic silica) concentration at site MD02-2529 recovered in the coastal Panama Basin. Piston core MD02-2529, collected in an area affected by a multitude of processes, provides evidence for strong variations in diatom production at the millennial timescale during the last glacial cycle. The maxima in total diatom concentration occurred during the early marine isotopic stage (MIS) 4 as well as during the MIS 4/3 transition and MIS 3. Rapid changes in diatom concentrations during the MIS 3 mimics Bond cycles as independently recorded by the SSS estimation derived from planktonic foraminifera from the same core. Such patterns indicate a clear linkage between diatom production in the coastal EEP and rapid climate changes in the high-latitude North Atlantic. In parallel, the long-term succession of the diatom community from coastal diatoms, predominantly thriving during MIS 5 and 4, towards pelagic diatoms, dominant during MIS 3 and 2, points to a long-term change in the surface hydrology. During Heinrich Events, diatoms strongly reduced their production, probably due to enhanced stratification in the upper water column. After the last glacial maximum, diatom production and valve preservation strongly decreased in response to the advection of nutrient (H2SiO4)-depleted, warmer water masses. Our high-resolution record highlights how regional climatic processes can modulate rapid changes in siliceous primary production as triggered by wind-induced local upwelling, indicating that millennial climatic variability can overtake other prominent hydrological processes such as those related to silicic acid leakage.

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Reflecting the natural biology of mass spawning fish aquaculture production of fish larvae is often hampered by high and unpredictable mortality rates. The present study aimed to enhance larval performance and immunity via the oral administration of an immunomodulator, beta-glucan (MacroGard®) in turbot (Scophthalmus maximus). Rotifers (Brachionus plicatilis) were incubated with or without yeast beta-1,3/1,6-glucan in form of MacroGard® at a concentration of 0.5 g/L. Rotifers were fed to first feeding turbot larvae once a day. From day 13 dph onwards all tanks were additionally fed untreated Artemia sp. nauplii (1 nauplius ml/L). Daily mortality was monitored and larvae were sampled at 11 and 24 dph for expression of 30 genes, trypsin activity and size measurements. Along with the feeding of beta-glucan daily mortality was significantly reduced by ca. 15% and an alteration of the larval microbiota was observed. At 11 dph gene expression of trypsin and chymotrypsin was elevated in the MacroGard® fed fish, which resulted in heightened tryptic enzyme activity. No effect on genes encoding antioxidative proteins was observed, whilst the immune response was clearly modulated by beta-glucan. At 11 dph complement component c3 was elevated whilst cytokines, antimicrobial peptides, toll like receptor 3 and heat shock protein 70 were not affected. At the later time point (24 dph) an anti-inflammatory effect in form of a down-regulation of hsp 70, tnf-alpha and il-1beta was observed. We conclude that the administration of beta-glucan induced an immunomodulatory response and could be used as an effective measure to increase survival in rearing of turbot.

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We present in situ microelectrode measurements of sediment formation factor and porewater oxygen and pH from six stations in the North Atlantic varying in depth from 2159 to 5380 m. A numerical model of the oxygen data indicates that fluxes of oxygen to the sediments are as much as an order of magnitude higher than benthic chamber flux measurements previously reported in the same area. Model results require dissolution driven by metabolic CO2 production within the sediments to explain the pH data; even at the station with the most undersaturated bottom waters >60% of the calcite dissolution occurs in response to metabolic CO2. Aragonite dissolution alone cannot provide the observed buffering of porewater pH, even at the shallowest station. A sensitivity test of the model that accounts for uncertainties in the bottom water saturation state and the stoichiometry between oxygen consumption and CO2 production during respiration constrains the dissolution rate constant for calcite to between 3 and 30% day**-1, in agreement with earlier in situ determinations of the rate constant. Model results predict that over 35% of the calcium carbonate rain to these sediments dissolves at all stations, confirmed by sediment trap and CaCO3 accumulation data.

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We examine the quantitative composition of benthic foraminiferal assemblages of Rose Bengal-stained surface samples from 37 stations in the Laptev Sea, and combine this data set with an existing data set along a transect from Spitsbergen to the central Arctic Ocean. Foraminiferal test accumulation rates, diversity, faunal composition and statistically defined foraminiferal associations are analysed for living (Rose Bengal-stained) and dead foraminifers. We compare the results of several benthic foraminiferal diversity indices and statistically defined foraminiferal associations, including Fisher's alpha and Shannon-Wiener diversity indices, Q-mode principal component analysis and correspondence analysis. Diversity and faunal density (standing stock) of living benthic foraminifers are positively correlated to trophic resources. In contrast, the accumulation rate of dead foraminifers (BFAR) shows fluctuating values depending on test disintegration processes. Foraminiferal associations defined by Q-mode principal component analysis and correspondence analysis are comparable. The factor values of the correspondence analysis allow a quantitative correlation between the foraminiferal fauna and the local carbon flux, which may be used as a tool to estimate changes in primary productivity.

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Bashōan Tōsei den / Uchida Roan [cho]: p. 240-300.