776 resultados para Negative Life Events
Resumo:
Palaeoecological evidence indicates that highland pines were dominant in extensive areas of the mountains of Central and Northern Iberia during the first half of the Holocene. However, following several millennia of anthropogenic pressure, their natural ranges are now severely reduced. Although pines have been frequently viewed as first-stage successional species responding positively to human disturbance, some recent palaeobotanical work has proposed fire disturbance and human deforestation as the main drivers of this vegetation turnover. To assess the strength of the evidence for this hypothesis and to identify other possible explanations for this scenario, we review the available information on past vegetation change in the mountains of northern inland Iberia. We have chosen data from several sites that offer good chronological control, including palynological records with microscopic charcoal data and sites with plant macro- and megafossil occurrence. We conclude that although the available long-term data are still fragmentary and that new methods are needed for a better understanding of the ecological history of Iberia, fire events and human activities (probably modulated by climate) have triggered the pine demise at different locations and different temporal scales. In addition, all palaeoxylological, palynological and charcoal results obtained so far are fully compatible with a rapid human-induced ecological change that could have caused a range contraction of highland pines in western Iberia.
Resumo:
The study of pointer years of numerous tree-ring chronologies of the central Iberian Peninsula (Sierra de Guadarrama) could provide complementary information about climate variability over the last 405 yr. In total, 64 pointer years have been identified: 30 negative (representing minimum growths) and 34 positive (representing maximum growths), the most significant of these being 1601, 1963 and 1996 for the negative ones, and 1734 and 1737 for the positive ones. Given that summer precipitation was found to be the most limiting factor for the growth of Pinus in the Sierra de Guadarrama in the second half of the 20th century, it is also an explanatory factor in almost 50% of the extreme growths. Furthermore, these pointer years and intervals are not evenly distributed throughout time. Both in the first half of the 17th and in the second half of 20th, they were more frequent and more extreme and these periods are the most notable for the frequency of negative pointer years in Central Spain. The interval 1600–1602 is of special significance, being one of the most unfavourable for tree growth in the centre of Spain, with 1601 representing the minimum index in the regional chronology. We infer that this special minimum annual increase was the effect of the eruption of Huaynaputina, which occurred in Peru at the beginning of 1600 AD. This is the first time that the effects of this eruption in the tree-ring records of Southern Europe have been demonstrated.
Resumo:
In the European context of upgrading the housing stock energy performance, multiple barriers hinder the wide uptake of sustainable retrofitting practices. Moreover, some of these may imply negative effects often disregarded. Policy makers need to identify how to increase and improve retrofitting practices from the comprehensive point of view of sustainability. None of the existing assessment tools addresses all the issues relevant for sustainable development in a local situation from a life cycle perspective. Life cycle sustainability assessment methodology, or LCSA, analyzes environmental and socioeconomic impacts. The environmental part is quite developed, but the socioeconomic aspect is still challenging. This work proposes socioeconomic criteria to be included in a LCSA to assess retrofitting works in the specific context of Brussels-Capital Region. LCSA feasibility and challenging methodology aspects are discussed.
Resumo:
Although tree ferns are an important component of temperate and tropical forests, very little is known about their ecology. Their peculiar biology (e.g., dispersal by spores and two-phase life cycle) makes it difficult to extrapolate current knowledge on the ecology of other tree species to tree ferns. In this paper, we studied the effects of negative density dependence (NDD) and environmental heterogeneity on populations of two abundant tree fern species, Cyathea caracasana and Alsophila engelii, and how these effects change across a successional gradient. Species patterns harbor information on processes such as competition that can be easily revealed using point pattern analysis techniques. However, its detection may be difficult due to the confounded effects of habitat heterogeneity. Here, we mapped three forest plots along a successional gradient in the montane forests of Southern Ecuador. We employed homogeneous and inhomogeneous K and pair correlation functions to quantify the change in the spatial pattern of different size classes and a case-control design to study associations between juvenile and adult tree ferns. Using spatial estimates of the biomass of four functional tree types (short- and long-lived pioneer, shade- and partial shade-tolerant) as covariates, we fitted heterogeneous Poisson models to the point pattern of juvenile and adult tree ferns and explored the existence of habitat dependencies on these patterns. Our study revealed NDD effects for C. caracasana and strong environmental filtering underlying the pattern of A. engelii. We found that adult and juvenile populations of both species responded differently to habitat heterogeneity and in most cases this heterogeneity was associated with the spatial distribution of biomass of the four functional tree types. These findings show the effectiveness of factoring out environmental heterogeneity to avoid confounding factors when studying NDD and demonstrate the usefulness of covariate maps derived from mapped communities.
Resumo:
La actividad volcánica interviene en multitud de facetas de la propia actividad humana, no siempre negativas. Sin embargo, son más los motivos de peligrosidad y riesgo que incitan al estudio de la actividad volcánica. Existen razones de seguridad que inciden en el mantenimiento del seguimiento y monitorización de la actividad volcánica para garantizar la vida y la seguridad de los asentamientos antrópicos en las proximidades de los edificios volcánicos. En esta tesis se define e implementa un sistema de monitorización de movimientos de la corteza en las islas de Tenerife y La Palma, donde el impacto social que representa un aumento o variación de la actividad volcánica en las islas es muy severo. Aparte de la alta densidad demográfica del Archipiélago, esta población aumenta significativamente, en diferentes periodos a lo largo del año, debido a la actividad turística que representa la mayor fuente de ingresos de las islas. La población y los centros turísticos se diseminan predominantemente a lo largo de las costas y también a lo largo de los flancos de los edificios volcánicos. Quizá el mantenimiento de estas estructuras sociales y socio-económicas son los motivos más importantes que justifican una monitorización de la actividad volcánica en las Islas Canarias. Recientemente se ha venido trabajando cada vez más en el intento de predecir la actividad volcánica utilizando los nuevos sistemas de monitorización geodésica, puesto que la actividad volcánica se manifiesta anteriormente por deformación de la corteza terrestre y cambios en la fuerza de la gravedad en la zona donde más tarde se registran eventos volcánicos. Los nuevos dispositivos y sensores que se han desarrollado en los últimos años en materias como la geodesia, la observación de la Tierra desde el espacio y el posicionamiento por satélite, han permitido observar y medir tanto la deformación producida en el terreno como los cambios de la fuerza de la gravedad antes, durante y posteriormente a los eventos volcánicos que se producen. Estos nuevos dispositivos y sensores han cambiado las técnicas o metodologías geodésicas que se venían utilizando hasta la aparición de los mismos, renovando métodos clásicos y desarrollando otros nuevos que ya se están afianzando como metodologías probadas y reconocidas para ser usadas en la monitorización volcánica. Desde finales de la década de los noventa del siglo pasado se han venido desarrollando en las Islas Canarias varios proyectos que han tenido como objetivos principales el desarrollo de nuevas técnicas de observación y monitorización por un lado y el diseño de una metodología de monitorización volcánica adecuada, por otro. Se presenta aquí el estudio y desarrollo de técnicas GNSS para la monitorización de deformaciones corticales y su campo de velocidades para las islas de Tenerife y La Palma. En su implementación, se ha tenido en cuenta el uso de la infraestructura geodésica y de monitorización existente en el archipiélago a fin de optimizar costes, además de complementarla con nuevas estaciones para dar una cobertura total a las dos islas. Los resultados obtenidos en los proyectos, que se describen en esta memoria, han dado nuevas perspectivas en la monitorización geodésica de la actividad volcánica y nuevas zonas de interés que anteriormente no se conocían en el entorno de las Islas Canarias. Se ha tenido especial cuidado en el tratamiento y propagación de los errores durante todo el proceso de observación, medida y proceso de los datos registrados, todo ello en aras de cuantificar el grado de fiabilidad de los resultados obtenidos. También en este sentido, los resultados obtenidos han sido verificados con otros procedentes de sistemas de observación radar de satélite, incorporando además a este estudio las implicaciones que el uso conjunto de tecnologías radar y GNSS tendrán en un futuro en la monitorización de deformaciones de la corteza terrestre. ABSTRACT Volcanic activity occurs in many aspects of human activity, and not always in a negative manner. Nonetheless, research into volcanic activity is more likely to be motivated by its danger and risk. There are security reasons that influence the monitoring of volcanic activity in order to guarantee the life and safety of human settlements near volcanic edifices. This thesis defines and implements a monitoring system of movements in the Earth’s crust in the islands of Tenerife and La Palma, where the social impact of an increase (or variation) of volcanic activity is very severe. Aside from the high demographic density of the archipelago, the population increases significantly in different periods throughout the year due to tourism, which represents a major source of revenue for the islands. The population and the tourist centres are mainly spread along the coasts and also along the flanks of the volcanic edifices. Perhaps the preservation of these social and socio-economic structures is the most important reason that justifies monitoring volcanic activity in the Canary Islands. Recently more and more work has been done with the intention of predicting volcanic activity, using new geodesic monitoring systems, since volcanic activity is evident prior to eruption because of a deformation of the Earth’s crust and changes in the force of gravity in the zone where volcanic events will later be recorded. The new devices and sensors that have been developed in recent years in areas such as geodesy, the observation of the Earth from space, and satellite positioning have allowed us to observe and measure the deformation produced in the Earth as well as the changes in the force of gravity before, during, and after the volcanic events occur. The new devices and sensors have changed the geodetic techniques and methodologies that were used previously. The classic methods have been renovated and other newer ones developed that are now vouched for as proven recognised methodologies to be used for volcanic monitoring. Since the end of the 1990s, in the Canary Islands various projects have been developed whose principal aim has been the development of new observation and monitoring techniques on the one hand, and the design of an appropriate volcanic monitoring methodology on the other. The study and development of GNSS techniques for the monitoring of crustal deformations and their velocity field is presented here. To carry out the study, the use of geodetic infrastructure and existing monitoring in the archipelago have been taken into account in order to optimise costs, besides complementing it with new stations for total coverage on both islands. The results obtained in the projects, which are described below, have produced new perspectives in the geodetic monitoring of volcanic activity and new zones of interest which previously were unknown in the environment of the Canary Islands. Special care has been taken with the treatment and propagation of errors during the entire process of observing, measuring, and processing the recorded data. All of this was done in order to quantify the degree of trustworthiness of the results obtained. Also in this sense, the results obtained have been verified with others from satellite radar observation systems, incorporating as well in this study the implications that the joint use of radar technologies and GNSS will have for the future of monitoring deformations in the Earth’s crust.
Resumo:
Acknowledgements The iHARP database was funded by unrestricted grants from Mundipharma International Ltd and Research in Real-Life Ltd; these analyses were funded by an unrestricted grant from Teva Pharmaceuticals. Mundipharma and Teva played no role in study conduct or analysis and did not modify or approve the manuscript. The authors wish to direct a special appreciation to all the participants of the iHARP group who contributed data to this study and to Mundipharma, sponsors of the iHARP group. In addition, we thank Julie von Ziegenweidt for assistance with data extraction and Anna Gilchrist and Valerie L. Ashton, PhD, for editorial assistance. Elizabeth V. Hillyer, DVM, provided editorial and writing support, funded by Research in Real-Life, Ltd.
Resumo:
The yeast Saccharomyces cerevisiae has a limited life-span, which is measured by the number of divisions that individual cells complete. Among the many changes that occur as yeasts age are alterations in chromatin-dependent transcriptional silencing. We have genetically manipulated histone deacetylases to modify chromatin, and we have examined the effect on yeast longevity. Deletion of the histone deacetylase gene RPD3 extended life-span. Its effects on chromatin functional state were evidenced by enhanced silencing at the three known heterochromatic regions of the genome, the silent mating type (HM), subtelomeric, and rDNA loci, which occurred even in the absence of SIR3. Similarly, the effect of the rpd3Δ on life-span did not depend on an intact Sir silencing complex. In fact, deletion of SIR3 itself had little effect on life-span, although it markedly accelerated the increase in cell generation time that is observed during yeast aging. Deletion of HDA1, another histone deacetylase gene, did not result in life-span extension, unless it was combined with deletion of SIR3. The hda1Δ sir3Δ resulted in an increase in silencing, but only at the rDNA locus. Deletion of RPD3 suppressed the loss of silencing in rDNA in a sir2 mutant; however, the silencing did not reach the level found in the rpd3Δ single mutant, and RPD3 deletion did not overcome the life-span shortening seen in the sir2 mutant. Deletion of both RPD3 and HDA1 caused a decrease in life-span, which resulted from a substantial increase in initial mortality of the population. The expression of both of these genes declines with age, providing one possible explanation for the increase in mortality during the life-span. Our results are consistent with the loss of rDNA silencing leading to aging in yeast. The functions of RPD3 and HDA1 do not overlap entirely. RPD3 exerts its effect on chromatin at additional sites in the genome, raising the possibility that events at loci other than rDNA play a role in the aging process.
Resumo:
We have investigated whether exposure to Gram-negative bacterial endotoxin in early neonatal life can alter neuroendocrine and immune regulation in adult animals. Exposure of neonatal rats to a low dose of endotoxin resulted in long-term changes in hypothalamic–pituitary–adrenal (HPA) axis activity, with elevated mean plasma corticosterone concentrations that resulted from increased corticosterone pulse frequency and pulse amplitude. In addition to this marked effect on the development of the HPA axis, neonatal endotoxin exposure had long-lasting effects on immune regulation, including increased sensitivity of lymphocytes to stress-induced suppression of proliferation and a remarkable protection from adjuvant-induced arthritis. These findings demonstrate a potent and long-term effect of neonatal exposure to inflammatory stimuli that can program major changes in the development of both neuroendocrine and immunological regulatory mechanisms.
Resumo:
Friend of GATA (FOG) proteins regulate GATA factor-activated gene transcription. During vertebrate hematopoiesis, FOG and GATA proteins cooperate to promote erythrocyte and megakaryocyte differentiation. The Drosophila FOG homologue U-shaped (Ush) is expressed similarly in the blood cell anlage during embryogenesis. During hematopoiesis, the acute myeloid leukemia 1 homologue Lozenge and Glial cells missing are required for the production of crystal cells and plasmatocytes, respectively. However, additional factors have been predicted to control crystal cell proliferation. In this report, we show that Ush is expressed in hemocyte precursors and plasmatocytes throughout embryogenesis and larval development, and the GATA factor Serpent is essential for Ush embryonic expression. Furthermore, loss of ush function results in an overproduction of crystal cells, whereas forced expression of Ush reduces this cell population. Murine FOG-1 and FOG-2 also can repress crystal cell production, but a mutant version of FOG-2 lacking a conserved motif that binds the corepressor C-terminal binding protein fails to affect the cell lineage. The GATA factor Pannier (Pnr) is required for eye and heart development in Drosophila. When Ush, FOG-1, FOG-2, or mutant FOG-2 is coexpressed with Pnr during these developmental processes, severe eye and heart phenotypes result, consistent with a conserved negative regulation of Pnr function. These results indicate that the fly and mouse FOG proteins function similarly in three distinct cellular contexts in Drosophila, but may use different mechanisms to regulate genetic events in blood vs. cardial or eye cell lineages.
Resumo:
The recently sequenced genome of the parasitic bacterium Mycoplasma genitalium contains only 468 identified protein-coding genes that have been dubbed a minimal gene complement [Fraser, C.M., Gocayne, J.D., White, O., Adams, M.D., Clayton, R.A., et al. (1995) Science 270, 397-403]. Although the M. genitalium gene complement is indeed the smallest among known cellular life forms, there is no evidence that it is the minimal self-sufficient gene set. To derive such a set, we compared the 468 predicted M. genitalium protein sequences with the 1703 protein sequences encoded by the other completely sequenced small bacterial genome, that of Haemophilus influenzae. M. genitalium and H. influenzae belong to two ancient bacterial lineages, i.e., Gram-positive and Gram-negative bacteria, respectively. Therefore, the genes that are conserved in these two bacteria are almost certainly essential for cellular function. It is this category of genes that is most likely to approximate the minimal gene set. We found that 240 M. genitalium genes have orthologs among the genes of H. influenzae. This collection of genes falls short of comprising the minimal set as some enzymes responsible for intermediate steps in essential pathways are missing. The apparent reason for this is the phenomenon that we call nonorthologous gene displacement when the same function is fulfilled by nonorthologous proteins in two organisms. We identified 22 nonorthologous displacements and supplemented the set of orthologs with the respective M. genitalium genes. After examining the resulting list of 262 genes for possible functional redundancy and for the presence of apparently parasite-specific genes, 6 genes were removed. We suggest that the remaining 256 genes are close to the minimal gene set that is necessary and sufficient to sustain the existence of a modern-type cell. Most of the proteins encoded by the genes from the minimal set have eukaryotic or archaeal homologs but seven key proteins of DNA replication do not. We speculate that the last common ancestor of the three primary kingdoms had an RNA genome. Possibilities are explored to further reduce the minimal set to model a primitive cell that might have existed at a very early stage of life evolution.
Resumo:
Many studies have characterized the transmembrane signaling events initiated after T-cell antigen receptor recognition of major histocompatibility complex (MHC)-bound peptides. Yet, little is known about signal transduction from a set of MHC class I recognizing receptors on natural killer (NK) cells whose ligation dramatically inhibits NK cell-mediated killing. In this study we evaluated the influence of MHC recognition on the proximal signaling events in NK cells binding tumor targets. We utilized two experimental models where NK cell-mediated cytotoxicity was fully inhibited by the recognition of specific MHC class I molecules. NK cell binding to either class I-deficient or class I-transfected target cells initiated rapid protein tyrosine kinase activation. In contrast, whereas NK cell binding to class I-deficient targets led to inositol phosphate release and increased intracellular free calcium ([Ca2+]i), NK recognition of class I-bearing targets did not induce the activation of these phospholipase C-dependent signaling events. The recognition of class I by NK cells clearly had a negative regulatory effect since blocking this interaction using anti-class I F(ab')2 fragments increased inositol 1,4,5-trisphosphate release and [Ca2+]i and increased the lysis of the targets. These results suggest that one of the mechanisms by which NK cell recognition of specific MHC class I molecules can block the development of cell-mediated cytotoxicity is by inhibiting specific critical signaling events.
Resumo:
This study explores the relationship between conformity to traditional feminine gender norms and meaning and purpose in life among female college students. Based on previous research findings that show a negative correlation between specific traditional feminine gender norm conformity factors and psychological well-being, we posited that participants with higher levels of traditional gender norm conformity will experience a lower sense of meaning and purpose in life. Self-report data was obtained from 338 female undergraduate college students using the Conformity to Feminine Norms Inventory (CFNI) and the Purpose in Life Test (PIL). A standard multiple regression assessed the accuracy of eight feminine norms in predicting purpose and meaning in life. Results partially supported the hypothesis, with Modesty as the most significant contributor to meaning and purpose. Inconsistent with our hypothesis, participants who endorsed high scores on Nice in Relationships, Involvement with Children, Sexual Fidelity, and Domestic, scored higher on the PIL.
Resumo:
Muitos são os benefícios provenientes da arborização de vias públicas, como aumento da vida útil do asfalto por meio do sombreamento, filtragem de poluentes, absorção de CO2, interceptação da água da chuva e da radiação solar e mitigação de ilhas de calor; porém, por questões culturais, é considerada muitas vezes pela população como algo negativo, cujas folhas entopem calhas, raízes destroem calçadas, troncos e folhas atrapalham fiação elétrica e, principalmente, elas estão susceptíveis às quedas. A pesquisa teve como objetivo estudar o comportamento das quedas de árvores no município de Piracicaba/SP, Brasil, com atenção especial ao regime de ventos na cidade, baseado em dados fornecidos pelo Corpo de Bombeiros de Piracicaba, Secretaria de Defesa do Meio Ambiente de Piracicaba (SEDEMA) e Estação Meteorológica da Escola Superior de Agricultura \"Luiz de Queiroz\" (ESALQ). De caráter inédito, o principal resultado foi a relação direta entre quedas e zonas urbanizadas, que, em constantes alterações no uso do solo, acabam por danificar a sustentação das árvores (raízes), fato justificado pela alta concentração de quedas na parte central (36,7%), com destaque para as estações da primavera e do verão, ou seja, ventos e chuvas, na qual, juntas, somaram 78,0% do total. O período de retorno esperado para ventos superiores a 75 km h-1, classificados como temporais na Escala de Vento de Beaufort, foi de 2,8 eventos por ano. Enquanto verificou-se o predomínio de ventos alísios de sudeste, pertencentes ao quarto quadrante, a direção das rajadas de vento teve maior variação e, assim, com predomínio daqueles ventos, é recomendável instalações de parques industriais nas zonas norte, noroeste e oeste, haja vista a importância de evitar que os poluentes adentrem a cidade. As espécies de maior vulnerabilidade foram: Pachira aquatica Aubl. (monguba), Handroanthus sp. (ipê roxo) e Tipuana tipu (Benth.) Kuntze (tipuana). Por meio de simulação microclimática computacional, utilizando o programa ENVI-met versão 3.1, em dois estudos de caso, sendo um em bairro residencial e outro na Praça José Bonifácio, foi possível identificar locais de maior atenção quanto às quedas de árvores, em função da rugosidade do local, capaz de alterar a velocidade e a direção do vento.
Resumo:
A percepção sobre as causas dos acontecimentos faz parte da vida cotidiana. Atribui-se causas aos eventos na busca de entendimento que permita prever, controlar e alterar resultados futuros. Questões como \"Por que não passei na prova?\" e \"Porque fui o primeiro colocado no vestibular?\" conduzem à percepção de uma causa explicativa, seja para o fracasso ou para o sucesso. A Teoria da Atribuição Causal tem sido examinada para compreender e explicar como as pessoas interpretam os determinantes de seu sucesso ou fracasso em situações de desempenho. As causas percebidas estarão relacionadas à percepção de cada indivíduo sobre o evento, o que não implica causalidade real, dado que a ação será efetivada de acordo com a percepção de cada indivíduo sobre o evento. A abordagem teórica utilizada nessa pesquisa foi a Teoria da Atribuição Causal, proposta por Bernard Weiner, no contexto educacional, com foco nas atribuições causais para sucesso e fracasso acadêmicos. Neste cenário, o objetivo principal da pesquisa foi identificar as causas percebidas como explicativas do desempenho acadêmico de estudantes do curso de Ciências Contábeis. Buscou-se também obter evidências e subsidiar a discussão sobre a relação entre o sucesso e o fracasso acadêmico, a modalidade de ensino, a autoestima e o perfil do estudante. Os dados foram coletados por meio de um questionário aplicado aos estudantes de Ciências Contábeis de duas Universidades Federais que oferecem o curso em duas modalidades de ensino (presencial e a distância) e 738 respostas válidas foram obtidas para análise. O questionário foi estruturado em três blocos (I - desempenho e causas percebidas, II - mensuração da autoestima e III - perfil do estudante). Os resultados apresentaram um perfil com idade média dos estudantes de 27,4 anos (34,27 na modalidade EaD e 24,87 na modalidade Presencial). A maioria dos estudantes (83%) exercia atividade remunerada (90% na EaD e 80% na presencial) e as mulheres representaram a maioria dos respondentes (62% na modalidade EaD e 58% na modalidade presencial). As causas internas, especificamente o esforço e a capacidade, foram mais indicadas como explicativas do sucesso acadêmico e as causas externas, especificamente a dificuldade da tarefa, a flexibilidade de horário e a influência negativa do professor, foram as mais indicadas como explicativas do fracasso acadêmico. Como os resultados apontam que os estudantes indicaram com frequência a própria capacidade para explicar o sucesso, pode-se admitir manifestação da tendência autoservidora, que contribui para a manutenção da autoestima, diante da influência positiva na motivação. Entre as causas do sucesso, a capacidade associou-se a um nível mais elevado de autoestima e a causa sorte associou-se a um nível mais baixo de autoestima. Entre as causas do fracasso, a dificuldade da tarefa associou-se ao nível mais baixo de autoestima. Uma análise geral permite observar que os estudantes dedicam pouco tempo aos estudos, atribuem sucesso principalmente a si mesmos e o fracasso a terceiros, e apresentam uma elevada autoestima associada principalmente ao sucesso atribuído à capacidade. Em futuras pesquisas, recomenda-se estudos pilotos, com objetivo de definir outras atribuições causais para elaboração de novos instrumentos de coleta de dados, por meio de abordagem metodológica qualitativa, que possam ampliar os achados e contribuir com a literatura.
Resumo:
I investigated effects of annual spring flows on the abundance of Rio Grande silvery minnow, red shiner, and fathead minnow within two segments of the Middle Rio Grande (i.e., above and below the Rio Puerco) using 12 years of data. Rio Grande silvery minnow abundance was positively related to high spring flows and negatively related to low spring flows. Conversely, the abundance of the red shiner and fathead minnow were negatively related to high spring flows and positively related to low spring flows. The strength of these relationships varied with species and location but results from analyses below the Rio Puerco showed a stronger positive and negative relationship (with high and low spring flows, respectively) for Rio Grande silvery minnow.