Mastogloia smithii var lacustris Grun.: A Structural Engineer of Calcareous Mats in Karstic Subtropical Wetlands

Mastogloia smithii var. lacustris Grun. is the dominant diatom in periphyton mats of the calcareous, freshwater to brackish wetlands of Caribbean coasts. Despite oligotrophy, frequent desiccation, high irradiance and temperatures, and occasional fire, periphyton communities in these wetlands can produce over 2000 g m-2 of organic biomass, prompting studies that examine stress resistance and maintenance of algal mats under extreme conditions. The diatom flora inhabiting periphyton mats from over 500 sites in the Florida Everglades and similar wetlands in Belize, Jamaica and Mexico was examined, and M. smithii var. lacustris was a persistent component, present in 97% of samples and comprising up to 80% of a diverse diatom assemblage. Valves at various stages of division were observed encased in extracellular polysaccharide that exceeded the cell volume; SEM observations confirm issuance from mantle pores resulting in suspension of the cell in a matrix dominated by cyanobacterial filaments. Using corresponding biophysical data from the collection sites, we define the optima for M. smithii var. lacustris along salinity, pH, phosphorus, and water depth gradients. Experiments revealed a collapse of M. smithii var. lacustris populations in the presence of above-ambient phosphorus concentrations and a rapid resurgence upon reflooding of desiccated mats. This widespread diatom taxon appears to play a critical role similar to that of cyanobacteria in microbial mats, and its disappearance in the presence of enrichment threatens biodiversity and the natural function in these systems that are increasingly influenced by urbanization

Tree Mortality following Prescribed Fire and a Storm Surge Event in Slash Pine (Pinus elliottii var. densa) Forests in the Florida Keys, USA

In fire-dependent forests, managers are interested in predicting the consequences of prescribed burning on postfire tree mortality. We examined the effects of prescribed fire on tree mortality in Florida Keys pine forests, using a factorial design with understory type, season, and year of burn as factors. We also used logistic regression to model the effects of burn season, fire severity, and tree dimensions on individual tree mortality. Despite limited statistical power due to problems in carrying out the full suite of planned experimental burns, associations with tree and fire variables were observed. Post-fire pine tree mortality was negatively correlated with tree size and positively correlated with char height and percent crown scorch. Unlike post-fire mortality, tree mortality associated with storm surge from Hurricane Wilma was greater in the large size classes. Due to their influence on population structure and fuel dynamics, the size-selective mortality patterns following fire and storm surge have practical importance for using fire as a management tool in Florida Keys pinelands in the future, particularly when the threats to their continued existence from tropical storms and sea level rise are expected to increase.

Pathogen-Specific Adaptations to Conserved Signaling Pathways in Cryptococcus neoformans

Cryptococcus neoformans is an opportunistic fungal pathogen that causes significant disease worldwide. Even though this fungus has not evolved specifically to cause human disease, it has a remarkable ability to adapt to many different environments within its infected host. C. neoformans adapts by utilizing conserved eukaryotic and fungal-specific signaling pathways to sense and respond to stresses within the host. Upon infection, two of the most significant environmental changes this organism experiences are elevated temperature and high pH.

Conserved Rho and Ras family GTPases are central regulators of thermotolerance in C. neoformans. Many GTPases require prenylation to associate with cellular membranes and function properly. Using molecular genetic techniques, microscopy, and infection models, I demonstrated that the prenyltransferase, geranylgeranyl transferase I (GGTase I) is required for thermotolerance and pathogenesis. Using fluorescence microscopy, I found that only a subset of conserved GGTase I substrates requires this enzyme for membrane localization. Therefore, the C. neoformans GGTase I may recognize its substrate in a slightly different manner than other eukaryotic organisms.

The alkaline response transcription factor, Rim101, is a central regulator of stress-response genes important for adapting to the host environment. In particular, Rim101 regulates cell surface alterations involved in immune avoidance. In other fungi, Rim101 is activated by alkaline pH through a conserved signaling pathway, but this pathway had yet been characterized in C. neoformans. Using molecular genetic techniques, I identified and analyzed the conserved members of the Rim pathway. I found that it was only partially conserved in C. neoformans, missing the components that sense pH and initiate pathway activation. Using a genetic screen, I identified a novel Rim pathway component named Rra1. Structural prediction and genetic epistasis experiments suggest that Rra1 may serve as the Rim pathway pH sensor in C. neoformans and other related basidiomycete fungi.

To explore the relevance of Rim pathway signaling in the interaction of C neoformans with its host, I characterized the Rim101-regulated cell wall changes that prevent immune detection. Using HPLC, enzymatic degradation, and cell wall stains, I found that the rim101Δ mutation resulted in increased cell wall chitin exposure. In vitro co-culture assays demonstrated that increased chitin exposure is associated with enhanced activation of macrophages and dendritic cells. To further test this association, I demonstrated that other mutant strains with increased chitin exposure induce macrophage and dendritic cell responses similar to rim101Δ. We used primary macrophages from mutant mouse lines to demonstrate that members of both the Toll-like receptor and C-type lectin receptor families are involved in detecting strains with increased chitin exposure. Finally, in vivo immunological experiments demonstrated that the rim101Δ strain induced a global inflammatory immune response in infected mouse lungs, expanding upon our previous in vivo rim101Δ studies. These results demonstrate that cell wall organization largely determines how fungal cells are detected by the immune system.

Relative Contributions of Prenylation and Postprenylation Processing in Cryptococcus neoformans Pathogenesis.

Prenyltransferase enzymes promote the membrane localization of their target proteins by directing the attachment of a hydrophobic lipid group at a conserved C-terminal CAAX motif. Subsequently, the prenylated protein is further modified by postprenylation processing enzymes that cleave the terminal 3 amino acids and carboxymethylate the prenylated cysteine residue. Many prenylated proteins, including Ras1 and Ras-like proteins, require this multistep membrane localization process in order to function properly. In the human fungal pathogen Cryptococcus neoformans, previous studies have demonstrated that two distinct forms of protein prenylation, farnesylation and geranylgeranylation, are both required for cellular adaptation to stress, as well as full virulence in animal infection models. Here, we establish that the C. neoformans RAM1 gene encoding the farnesyltransferase β-subunit, though not strictly essential for growth under permissive in vitro conditions, is absolutely required for cryptococcal pathogenesis. We also identify and characterize postprenylation protease and carboxyl methyltransferase enzymes in C. neoformans. In contrast to the prenyltransferases, deletion of the genes encoding the Rce1 protease and Ste14 carboxyl methyltransferase results in subtle defects in stress response and only partial reductions in virulence. These postprenylation modifications, as well as the prenylation events themselves, do play important roles in mating and hyphal transitions, likely due to their regulation of peptide pheromones and other proteins involved in development. IMPORTANCE Cryptococcus neoformans is an important human fungal pathogen that causes disease and death in immunocompromised individuals. The growth and morphogenesis of this fungus are controlled by conserved Ras-like GTPases, which are also important for its pathogenicity. Many of these proteins require proper subcellular localization for full function, and they are directed to cellular membranes through a posttranslational modification process known as prenylation. These studies investigate the roles of one of the prenylation enzymes, farnesyltransferase, as well as the postprenylation processing enzymes in C. neoformans. We demonstrate that the postprenylation processing steps are dispensable for the localization of certain substrate proteins. However, both protein farnesylation and the subsequent postprenylation processing steps are required for full pathogenesis of this fungus.

Influence of LAR and VAR on Para-Aminopyridine Antimalarials Targetting Haematin in Chloroquine-Resistance

Antimalarial chloroquine (CQ) prevents haematin detoxication when CQ-base concentrates in the acidic digestive vacuole through protonation of its p-aminopyridine (pAP) basic aro- matic nitrogen and sidechain diethyl-N. CQ export through the variant vacuolar membrane export channel, PFCRT, causes CQ-resistance in Plasmodium falciparum but 3-methyl CQ (sontochin SC), des-ethyl amodiaquine (DAQ) and bis 4-aminoquinoline piperaquine (PQ) are still active. This is determined by changes in drug accumulation ratios in parasite lipid (LAR) and in vacuolar water (VAR). Higher LAR may facilitate drug binding to and blocking PFCRT and also aid haematin in lipid to bind drug. LAR for CQ is only 8.3; VAR is 143,482. More hydrophobic SC has LAR 143; VAR remains 68,523. Similarly DAQ with a phenol sub- stituent has LAR of 40.8, with VAR 89,366. In PQ, basicity of each pAP is reduced by distal piperazine N, allowing very high LAR of 973,492, retaining VAR of 104,378. In another bis quinoline, dichlorquinazine (DCQ), also active but clinically unsatisfactory, each pAP retains basicity, being insulated by a 2-carbon chain from a proximal nitrogen of the single linking piperazine. While LAR of 15,488 is still high, the lowest estimate of VAR approaches 4.9 million. DCQ may be expected to be very highly lysosomotropic and therefore potentially hepatotoxic. In 11 pAP antimalarials a quadratic relationship between logLAR and logRe- sistance Index (RI) was confirmed, while log (LAR/VAR) vs logRI for 12 was linear. Both might be used to predict the utility of structural modifications.

Efectos de la temperatura sobre el peso seco, concentración de aceite y la proporción de ácidos grasos en el aceite de frutos de olivo (Olea europaea L. var Arauco)

Las respuestas de los componentes del rendimiento y de la calidad del aceite a la temperatura aún no han sido estudiadas en olivo ni en otras oleaginosas que acumulan aceite principalmente en mesocarpo (cerca de 95 por ciento del total en el caso de olivo). La información disponible en olivo se basa en correlaciones ligadas a variaciones en ubicación geográfica, altitud y años en los que pudieron covariar con la temperatura otros factores (radiación, disponibilidad hídrica, nutrición, etc). El objetivo de esta tesis fue evaluar el rol de la temperatura durante el crecimiento de frutos de olivo sobre el peso seco de los mismos, su contenido y concentración de aceite y las proporciones de ácidos grasos del aceite. Se realizaron experimentos manipulativos aplicados a ramas fructíferas creciendo a campo, utilizando cámaras transparentes con control y registro de temperatura. Los rangos de temperaturas medias alcanzados (entre 15-32 ºC) fueron mucho mayores a los rangos de 1 y 4 ºC logrados en estudios correlativos y permitieron generar relaciones funcionales para el peso seco del fruto, su concentración de aceite, y la proporción de ácidos grasos en el aceite. El contraste entre aplicaciones de temperaturas mayores o menores al ambiente durante períodos largos (114 días) y cortos (30 días) demostraron que tanto la proporción de aceite del fruto como la de ácido oleico en el aceite de fruto entero (mesocarpo + semilla) disminuye al aumentar la temperatura. Si bien en ambas estructuras del fruto la temperatura tuvo un efecto negativo sobre la concentración de aceite, las respuestas a la temperatura de la proporción de ácidos grasos de la semilla y el mesocarpo, medidos en forma separada, difirieron entre sí. Los patrones de respuesta de las proporciones de ácidos grasos en el aceite de semilla mostraron similitudes con las conocidas para especies oleaginosas de semilla como girasol. Por contraste, en el aceite de mesocarpo de olivo la proporción de ácido oleico disminuyó con el aumento de la temperatura, patrón opuesto a lo manifestado en semillas de olivo y en las de oleaginosas anuales. En esta tesis también se evaluó el efecto de la temperatura mínima nocturna sobre el crecimiento de los frutos y la calidad del aceite. Los resultados sugieren que la temperatura mínima nocturna y la amplitud térmica diarias son las dimensiones del régimen térmico diario con los que mejor se asociaron los cambios en el peso seco del fruto, la proporción de aceite y las proporciones de ácidos grasos en el mismo. Los resultados de esta tesis sirven para guiar la selección de sitios de plantación de nuevos olivares basados en los registros térmicos zonales, y para explicar los desajustes de la proporción de ácidos grasos del aceite de oliva producido en el NOA y la normativa del Consejo Oleícola Internacional (COI). Las relaciones funcionales entre las variables respuesta y la temperatura definidas podrán incorporarse en modelos de simulación del rendimiento y calidad que se desarrollen en el futuro. Tomados en conjunto con la tendencia al aumento de la temperatura, producto del calentamiento global, los resultados permitirán estimar la magnitud de los impactos negativos que podrían afectar el rendimiento del cultivo y la calidad del aceite.

Do U.S. and Colombian macro factors improve the forecasting ability of unrestricted VAR models of the local term structure of interest rate?

Este estudio empírico compara la capacidad de los modelos Vectores auto-regresivos (VAR) sin restricciones para predecir la estructura temporal de las tasas de interés en Colombia -- Se comparan modelos VAR simples con modelos VAR aumentados con factores macroeconómicos y financieros colombianos y estadounidenses -- Encontramos que la inclusión de la información de los precios del petróleo, el riesgo de crédito de Colombia y un indicador internacional de la aversión al riesgo mejora la capacidad de predicción fuera de la muestra de los modelos VAR sin restricciones para vencimientos de corto plazo con frecuencia mensual -- Para vencimientos de mediano y largo plazo los modelos sin variables macroeconómicas presentan mejores pronósticos sugiriendo que las curvas de rendimiento de mediano y largo plazo ya incluyen toda la información significativa para pronosticarlos -- Este hallazgo tiene implicaciones importantes para los administradores de portafolios, participantes del mercado y responsables de las políticas

Predicción de tipos de cambio reales utilizando modelos VAR Bayesianos

El presente trabajo busca documentar el potencial de los modelos VAR Bayesianos (BVAR) para la predicción de índices de tipos de cambio reales efectivos. Para esto, se prueban distintas especificaciones de modelos predictivos utilizando la base angosta de índices de tipos de cambio reales efectivos de BIS que incluye datos para 27 economías. En primera instancia se prueban modelos univariados simples para realizar las predicciones y tener un punto de referencia para las estimaciones BVAR. El análisis de los resultados de las predicciones de los modelos BVAR tradicionales muestran que estos por sí solos no tienen un mejor desempeño que los modelos univariados. Estos resultados son robustos a la ventana de estimación, y a la especificación de los priors del BVAR.

DINÂMICA DA REGENERAÇÃO NATURAL NO SUB-BOSQUE DE Pinus caribaea Morelet. var. caribaea NA RESERVA BIOLÓGICA DE SALTINHO, TAMANDARÉ - PE

O estudo da estrutura e dinâmica da regeneração natural em sub-bosque de plantios com espécies exóticas, como as do gênero Pinus , possibilita dar informações para manejo, conservação e reestabelecimento das espécies nativas de uma comunidade vegetal. O objetivo deste trabalho foi identificar e quantificar a dinâmica da regeneração natural das espécies arbustivo-arbóreas ocorrentes no sub-bosque do povoamento de Pinus caribaea , na Rebio de Saltinho, em Pernambuco. Foram medidas as espécies regenerantes de 10 parcelas permanentes, de 1 x 50 m, e incluídos os indivíduos com circunferência na base a 30 cm do solo (CAB 0,30m) ≤ 15 cm e altura superior a um metro. A altura foi classificada em: Classe 1, indivíduos arbustivoarbóreos, com altura 1 ≤ H ≤ 2; Classe 2 com altura 2 < H ≤ 3; e Classe 3, com altura > 3 m e CAP ≤ 15 cm. Calcularam-se os parâmetros fitossociológicos, a dinâmica da regeneração e os índices de Shannon (H’) e a equabilidade (J’) por Pielou. Protium heptaphyllum teve maior número de indivíduos e valor de importância (VI), e Miconia prasina a melhor frequência nos dois levantamentos. Quanto ao índice H’ de 3,32 nats.ind-1 (2007) passou a 3,07 nats.ind-1 (2012), e a equabilidade de J’ de 0,85 a 0,62, havendo decréscimo tanto para a diversidade, quanto para a distribuição. O levantamento de 2012 registrou aumento de 12,5% do número de indivíduos, e os regenerantes de 2007 tiveram 48,31% de mortalidade. Com relação ao número de indivíduos e área basal, os percentuais de ganhos foram superiores ao das perdas. Conclui-se que a sucessão ecológica da regeneração do sub-bosque do povoamento estudado, encontra-se em modificação positiva, e o povoamento de Pinus caribaea, não está impedindo o surgimento de novos indivíduos e espécies.

Efeito da salinidade da água do mar no rendimento, composição e atividades biológicas de frações polissacarídicas da Chlorophyta Caulerpa cupressoides var. flabellata

Seaweeds sulfated polysaccharides have been described as having various pharmacological activities. However, nothing is known about the influence of salinity on the structure of sulfated polysaccharides from green seaweed and pharmacological activities they perform. Therefore, the main aim of this study was to evaluate the effect of salinity of seawater on yield and composition of polysaccharides-rich fractions from green seaweed Caulerpa cupressoides var. flabellata, collected in two different salinities beaches of the coast of Rio Grande do Norte, and to verify the influence of salinity on their biological activities. We extracted four sulfated polysaccharides-rich fractions from C. cupressoides collected in Camapum beach (denominated CCM F0.3; F0.5; F1.0; F2.0), which the seawater has higher salinity, and Buzios beach (denominated CCB F0.3; F0.5; F1.0; F2.0). Different from that observed for other seaweeds, the proximate composition of C. cupressoides did not change with increased salinity. Moreover, interestingly, the C. cupresoides have high amounts of protein, greater even than other edible seaweeds. There was no significant difference (p>0.05) between the yield of polysaccharide fractions of CCM and its CCB counterparts, which indicates that salinity does not interfere with the yield of polysaccharide fractions. However, there was a significant difference in the sulfate/sugar ratio of F0.3 (p<0.05) and F0.5 (p<0.01) (CCM F0.3 and CCB F0.5 was higher than those determined for their counterparts), while the sulfate/sugar ratio the F1.0 and F2.0 did not change significantly (p>0.05) with salinity. This result suggested that the observed difference in the sulfate/sugar ratio between the fractions from CCM and CCB, is not merely a function of salinity, but probably also is related to the biological function of these biopolymers in seaweed. In addition, the salinity variation between collection sites did not influence algal monosaccharide composition, eletrophoretic mobility or the infrared spectrum of polysaccharides, demonstrating that the salinity does not change the composition of sulfated polysaccharides of C. cupressoides. There were differences in antioxidant and anticoagulant fractions between CCM and CCB. CCB F0.3 (more sulfated) had higher total antioxidant capacity that CCM F0.3, since the chelating ability the CCM F0.5 was more potent than CCB F0.5 (more sulfated). These data indicate that the activities of sulfated polysaccharides from CCM and CCB depend on the spatial patterns of sulfate groups and that it is unlikely to be merely a charge density effect. C. cupressoides polysaccharides also exhibited anticoagulant activity in the intrinsic (aPTT test) and extrinsic pathway (PT test). CCB F1.0 and CCM F1.0 showed different (p<0,001) aPTT activity, although F0.3 and F0.5 showed no difference (p>0,05) between CCM and CCB, corroborating the fact that the sulfate/sugar ratio is not a determining factor for biological activity, but rather for sulfate distribution along the sugar chain. Moreover, F0.3 and F0.5 activity in aPTT test was similar to that of clexane®, anticoagulant drug. In addition, F0.5 showed PT activity. These results suggest that salinity may have created subtle differences in the structure of sulfated polysaccharides, such as the distribution of sulfate groups, which would cause differences in biological activities between the fractions of the CCM and the CCB