Regional Risk Assessment of a Brazilian Rain Forest Reserve

The objective of this study was to identify subareas inside and near an Atlantic Rain Forest reserve, the Parque Estadual Turístico do Alto Ribeira (PETAR), most likely to be affected by land use in the vicinity of the area. In addition, the study aimed to compare risks per stressor source (agriculture, human settlements and mining) to both epigean (surface) and hypogean (subterranean) aquatic fauna. The methodological approach included the relative vulnerability of endpoints to the stressors (pesticides, metals, nutrients, and particles) and ranking of stressor sources and habitats (epigean and hypogean streams) based on their relative distribution in 14 subareas within the catchment areas of the main rivers that cross PETAR: Pilões, Betari and Iporanga. Four subareas presented high risk for both epigean and hypogean fauna. Three of those areas were located inside the Betari catchment area, where most of the settlements and abandoned lead mines are located. The fourth area was situated in the headwaters of the Pilões River, where agricultural activities are intense. Agriculture and human settlements were the activities most likely to cause impacts on aquatic ecosystems. Uses of risk assessment results include management of the PETAR and communication to stakeholders by the Park Administration.     

Recovery and purification of aprotinin from industrial insulin-processing effluent by immobilized chymotrypsin and negative IMAC chromatographies

Aprotinin, a bovine protease inhibitor currently also produced in recombinant bacteria, yeast, and corn, has valuable applications as a human therapeutic and in tissue culture. The objective of this work was to develop the basis of a large-scale aprotinin purification process centered on immobilized metal ion affinity chromatography (IMAC). This technique uses ligands—metal ions—of a lower cost and higher stability than those traditionally used in affinity chromatography. Since aprotinin does not interact with IMAC ligands, collection is from the nonretained fractions (negative chromatography). Stirred-tank batch IMAC adsorption experiments indicated that one-step aprotinin purification could not be successful. Immobilized chymotrypsin chromatography was then used as a prepurification step, yielding a suitable feed for IMAC (with purification factors as high as 476). IMAC column fed with these prepurified materials produced purified aprotinin in the nonretained fractions with purification factors as high as 952.     

Phosphatidylinositol 3-kinase

Currently, a central question in biology is how signals from the cell surface modulate intracellular processes. In recent years phosphoinositides have been shown to play a key role in signal transduction. Two phosphoinositide pathways have been characterized, to date. In the canonical phosphoinositide turnover pathway, activation of phosphatidylinositol-specific phospholipase C results in the hydrolysis of phosphatidylinositol 4,5-bisphospate and the generation of two second messengers, inositol 1,4,5-trisphosphate and diacylglycerol. The 3-phosphoinositide pathway involves protein-tyrosine kinase-mediated recruitment and activation of phosphatidylinositol 3-kinase, resulting in the production of phosphatidylinositol 3,4-bisphosphate and phosphatidylinositol 3,4,5-trisphosphate. The 3-phosphoinositides are not substrates of any known phospholipase C, are not components of the canonical phosphoinositide turnover pathway, and may themselves act as intracellular mediators. The 3-phosphoinositide pathway has been implicated in growth factor-dependent mitogenesis, membrane ruffling and glucose uptake. Furthermore the homology of the yeast vps34 with the mammalian phosphatidylinositol 3-kinase has suggested a role for this pathway in vesicular trafficking. In this review the different mechanisms employed by protein-tyrosine kinases to activate phosphatidylinositol 3-kinase, and its involvement in the signaling cascade initiated by tyrosine phosphorylation, are examined.     

Predicting impacts of lionfish (<Emphasis Type="Italic">Pterois volitans</Emphasis>) invasion in a coastal ecosystem of southern Brazil

The opportunistic feeder Pterois volitans is a voracious invader, causing large impacts in marine food-webs. We have used a Ecopath-with-Ecosim model to hypothesize an invasion by lionfish and to predict the likely impact of this potential generalist mesopredator in a subtropical food-web model. With thirty-three functional groups, the initial Ecopath model was balanced with a low biomass of lionfish (0.07 t/km²). In Ecosim, three scenarios of different vulnerability settings for the linkages between the introduced fish and its prey were tested, representing the default setting, a top-down control, and an extreme top-down control. The scenarios were tested using different assumptions on the ability of the invasive fish to change the proportions of prey consumed according to prey availability. Our model predicted that the hypothesized lionfish invasion would have a strong impact on this subtropical marine food web: (1) by reducing prey populations and, consequently, food for native predators, and; (2) by predating on key species, causing direct impacts and possibly cascading trophic effects. Reef fish were the most affected, including some groups ecologically and economically important, like lutjanids and groupers. However, some adaptations in the fishing strategy of fishermen are expected which may affect other fish groups. Stakeholders should be warned of the potential ecological and socio-economic impacts that may arise from a lionfish invasion and various strategies and policy options should be immediately developed and applied (1) to prevent the arrival and establishment of the lionfish, and; (2) to make the ecosystem more resilient to this and other possible exotic species.     

Brassinosteroid increases the cytokinin efficiency to induce direct somatic embryogenesis in leaf explants of <Emphasis Type="Italic">Coffea arabica</Emphasis> L. (Rubiaceae)

The combination of different plant growth regulators can result in beneficial effects in the induction of in vitro morphogenetic pathways. The present study reports the effect of 24-epibrassinolid (24-epiBR; brassinosteroid) when added alone and in association with N⁶-(2-isopentnyl) adenine (2-iP; cytokinin) in the induction of direct somatic embryogenesis in Coffea arabica. Leaf explants were cultivated in a modified Murashige and Skoog (MS) medium with 0 or 10 µM 2-iP and different concentrations (0.01, 0.10 or 1.0 µM) of 24-epiBR. Explants cultured on MS medium supplemented with 1.0 µM 24-epiBR in association with 2-iP produced 6.8 times more somatic embryos than the explants cultured with only 2-iP. Histological analyses also provided evidence that the supplementation of brassinosteroids in the culture medium could have influenced somatic embryogenesis differentiation. Somatic embryos obtained in the presence of brassinosteroid and cytokinin were better structured morpho-histologically as compared to those obtained in the medium with just cytokinin. This study opens new perspectives for the use of brassinosteroids in the somatic embryogenesis of C. arabica, so as to optimize the in vitro regeneration systems used in genetic improvement programs in C. arabica productive systems.     

The specificity of alcohol dehydrogenase with cis-retinoids. Activity with 11-cis-retinol and localization in retina.

Studies in knockout mice support the involvement of alcohol dehydrogenases ADH1 and ADH4 in retinoid metabolism, although kinetics with retinoids are not known for the mouse enzymes. Moreover, a role of alcohol dehydrogenase (ADH) in the eye retinoid interconversions cannot be ascertained due to the lack of information on the kinetics with 11-cis-retinoids. We report here the kinetics of human ADH1B1, ADH1B2, ADH4, and mouse ADH1 and ADH4 with all-trans-, 7-cis-, 9-cis-, 11-cis- and 13-cis-isomers of retinol and retinal. These retinoids are substrates for all enzymes tested, except the 13-cis isomers which are not used by ADH1. In general, human and mouse ADH4 exhibit similar activity, higher than that of ADH1, while mouse ADH1 is more efficient than the homologous human enzymes. All tested ADHs use 11-cis-retinoids efficiently. ADH4 shows much higher k(cat)/K(m) values for 11-cis-retinol oxidation than for 11-cis-retinal reduction, a unique property among mammalian ADHs for any alcohol/aldehyde substrate pair. Docking simulations and the kinetic properties of the human ADH4 M141L mutant demonstrated that residue 141, in the middle region of the active site, is essential for such ADH4 specificity. The distinct kinetics of ADH4 with 11-cis-retinol, its wide specificity with retinol isomers and its immunolocalization in several retinal cell layers, including pigment epithelium, support a role of this enzyme in the various retinol oxidations that occur in the retina. Cytosolic ADH4 activity may complement the isomer-specific microsomal enzymes involved in photopigment regeneration and retinoic acid synthesis.     

Bleomycin-induced pulmonary fibrosis is attenuated by a monoclonal antibody targeting HER2.

The importance of HER2/HER3 signaling in decreasing the effects of lung injury was recently demonstrated. Transgenic mice unable to signal through HER2/HER3 had significantly less bleomycin-induced pulmonary fibrosis and showed a survival benefit. Based on these data, we hypothesized that pharmacological blockade of HER2/HER3 in vivo in wild-type mice would have the same beneficial effects. We tested this hypothesis in a bleomycin lung injury model using 2C4, a monoclonal antibody directed against HER2 that blocks HER2/HER3 signaling. The administration of 2C4 before injury decreased the effects of bleomycin at days 15 and 21 after injury. HER2/HER3 blockade resulted in less collagen deposition (362.8 +/- 37.9 compared with 610.5 +/- 27.1 microg/mg; P = 0.03) and less lung morphological changes (injury score of 1.99 +/- 1.55 vs. 3.90 +/- 0.76; P < 0.04). In addition, HER2/HER3 blockade resulted in a significant survival advantage with 50% vs. 25% survival at 30 days (P = 0.04). These results confirm that HER2 signaling can be pharmacologically targeted to reduce lung fibrosis and remodeling after injury.     

The carboxylic acid transporters Jen1 and Jen2 affect the architecture and fluconazole susceptibility of Candida albicans biofilm in the presence of lactate

Candida albicans has the ability to adapt to different host niches, often glucose-limited but rich in alternative carbon sources. In these glucose-poor microenvironments, this pathogen expresses JEN1 and JEN2 genes, encoding carboxylate transporters, which are important in the early stages of infection. This work investigated how host microenvironments, in particular acidic containing lactic acid, affect C. albicans biofilm formation and antifungal drug resistance. Multiple components of the extracellular matrix were also analysed, including their impact on antifungal drug resistance, and the involvement of both Jen1 and Jen2 in this process. The results show that growth on lactate affects biofilm formation, morphology and susceptibility to fluconazole and that both Jen1 and Jen2 might play a role in these processes. These results support the view that the adaptation of Candida cells to the carbon source present in the host niches affects their pathogenicity.     

A New Depigmenting‐Antifungal Methylated Grindelane from Grindelia chiloensis

The new methylated grindelane diterpenoid, 7β ‐hydroxy‐8(17)‐dehydrogrindelic acid ( 1b ), together with the known 7α ‐hydroxy‐8(17)‐dehydrogrindelic acid ( 2a ), 6‐oxogrindelic acid ( 3a ), 4β ‐hydroxy‐6‐oxo‐19‐norgrindelic ( 4a ), 19‐hydroxygrindelic acid ( 5a ), 18‐hydroxygrindelic acid ( 6a ), 4α ‐carboxygrindelic acid ( 7a ), 17‐hydroxygrindelic acid ( 8a ), 6α ‐hydroxygrindelic acid ( 9a ), 8,17‐bisnor‐8‐oxagrindelic acid ( 10a ), 7α ,8α ‐epoxygrindelic acid ( 11a ), and strictanonic acid ( 12a ) as methyl esters were obtained from an Argentine collection of Grindelia chiloensis (Cornel .) Cabrera . Their structures and relative configurations were established on the basis of spectroscopic analysis. CHC l₃ extract from the aerial parts and their pure compounds were evaluated for their antifungal and depigmenting effects. Methyl ester derivative of 10a ( 10b ) exhibited a remarkable mycelial growth inhibition against Botritis cinerea with an IC ₅₀ of 13.5 μg ml^?1. While the new grindelane 1b exerted a clear color reduction of the yellow‐orange pigment developed by Fusarium oxysporum against UV ‐induced damage.