Rac, membrane heterogeneity, caveolin and regulation of growth by integrins

Anchorage dependence of growth blocks cell proliferation in inappropriate environments, thereby inhibiting cancer cell invasion and metastasis. Inhibition of growth regulatory pathways, including Rac, Erk and PtdIns 3-kinase in non-adherent cells mediates this effect. Here, we review recent work showing that integrin-mediated adhesion controls Rac binding to membranes. Rac binding sites can be found within cholesterol-enriched membrane domains, which are internalized when cells are deprived of adhesion. Endocytosis of these domains is mediated by caveolae and regulated by caveolin-1 phosphorylated on Tyr 14. This mechanism can account for the control of multiple pathways by integrins, thus providing an important mechanism for anchorage dependence of growth.     

Physiological and phylogenetic variability of Mexican <Emphasis Type="Italic">Metarhizium</Emphasis> strains

Metarhizium Sorokin is a wide spread entomopathogenic fungal genus that is also a root symbiont and some isolates may be potentially useful in agriculture applications. Physiological variations applicable in biological control have not been fully evaluated in Mexican strains. This study identified 30 strains from different insect hosts and states of Mexico selected because they represent the main sugar cane and corn plantations of the country. Their thermotolerance profiles (8, 15, 28 and 35 °C), insect virulence (LT50 in Galleria mellonella (Linnaeus) (Lepidoptera: Pyralidae) larvae), possible association with plant roots, and phylogenetic placement (using 5′TEF, IGS sequence information, and RFLP analysis) were assessed. Results showed no cold-active strains and most showed restricted growth at 35 °C. Insect virulence bioassays against G. mellonella showed highly virulent strains. Most of the strains showed an ability to associate with roots of Phaseolus vulgaris (Burkart) (Fabales: Fabaceae). RFLP analysis with 5′TEF and Pr1 revealed three distinct genetic groups. Phylogenetic analysis identified the 30 strains as Metarhizium anisopliae. (Metchnikoff) Sorokin, M. robertsii (J.F. Bisch., Rehner & Humber), M. guizhouense (Q. T. Chen & H. L. Guo), or M. pinghaense (Q.T. Chen & H.L. Guo), as well as two strains that belong to a new clade (Metarhizium sp. indet. 1). These data demonstrate the complexity of Metarhizium species and the need to study their physiological and phylogenetic aspects for a more adequate assessment of their potential use in agricultural systems.     

Two novel RAD21 mutations in patients with mild Cornelia de Lange syndrome-like presentation and report of the first familial case

Cornelia de Lange syndrome (CdLS) is a developmental disorder characterized by limb reduction defects, characteristic facial features and impaired cognitive development. Mutations in the NIPBL gene predominate; however, mutations in other cohesin complex genes have also been implicated, particularly in atypical and mild CdLS cases. Missense mutations and whole gene deletions in RAD21 have been identified in children with growth retardation, minor skeletal anomalies and facial features that overlap findings in individuals with CdLS. We report the first intragenic deletion and frameshift mutations identified in RAD21 in two patients presenting with atypical CdLS. One patient had an in-frame deletion of exon 13, while the second patient had a c.592_593dup frameshift mutation. The first patient presented with developmental delay, hypospadias, inguinal hernia and dysmorphic features while, the second patient presented with developmental delay, characteristic facial features, hirsutism, and hand and feet anomalies, with the first patient being milder than the second. The in-frame deletion mutation was found to be inherited from the mother who had a history of melanoma and other unspecified medical problems.     

Potential impacts of climate change on habitat suitability of Fagus sylvatica L. forests in spain

Abstract

Fagus sylvatica forests are considered to be of Community interest according to Directive 92/43/EEC. Climate change predictions for Spain point to a warming scenario, coupled with decreasing rainfall, which may have an impact on their future distribution particularly at the extremes of its distribution area. Species distribution models incorporating bioclimatic, topographic and phytogeographic variables were used as predictors to assess their habitat suitability under current conditions and a climate change projection. Ten single models were generated and an ensemble-forecasting model was subsequently built by computing a consensus of single-model projections. The results revealed that ombrothermic indices are the main factors controlling the distribution of Spanish beech forests. They are highly vulnerable to climate change and could suffer a decline in their habitat suitability if climate trends observed are maintained in the future. The least favoured areas for them will be located close to the limit between the Temperate and Mediterranean climates, where they could suffer a loss of habitat suitability. Conversely, suitable new areas could be found mainly in western areas of the Cantabrian Range and in the Central Pyrenees.     

Prolonged calcium influx after termination of light-induced calcium release in invertebrate photoreceptors

In microvillar photoreceptors, light stimulates the phospholipase C cascade and triggers an elevation of cytosolic Ca²⁺ that is essential for the regulation of both visual excitation and sensory adaptation. In some organisms, influx through light-activated ion channels contributes to the Ca²⁺ increase. In contrast, in other species, such as Lima, Ca²⁺ is initially only released from an intracellular pool, as the light-sensitive conductance is negligibly permeable to calcium ions. As a consequence, coping with sustained stimulation poses a challenge, requiring an alternative pathway for further calcium mobilization. We observed that after bright or prolonged illumination, the receptor potential of Lima photoreceptors is followed by the gradual development of an after-depolarization that decays in 1–4 minutes. Under voltage clamp, a graded, slow inward current (I_slow) can be reproducibly elicited by flashes that saturate the photocurrent, and can reach a peak amplitude in excess of 200 pA. I_slow obtains after replacing extracellular Na⁺ with Li⁺, guanidinium, or N-methyl-d-glucamine, indicating that it does not reflect the activation of an electrogenic Na/Ca exchange mechanism. An increase in membrane conductance accompanies the slow current. I_slow is impervious to anion replacements and can be measured with extracellular Ca²⁺ as the sole permeant species; Ba can substitute for Ca²⁺ but Mg²⁺ cannot. A persistent Ca²⁺ elevation parallels I_slow, when no further internal release takes place. Thus, this slow current could contribute to sustained Ca²⁺ mobilization and the concomitant regulation of the phototransduction machinery. Although reminiscent of the classical store depletion–operated calcium influx described in other cells, I_slow appears to diverge in some significant aspects, such as its large size and insensitivity to {"type":"entrez-protein","attrs":{"text":"SKF96365","term_id":"1156357400","term_text":"SKF96365"}}SKF96365 and lanthanum; therefore, it may reflect an alternative mechanism for prolonged increase of cytosolic calcium in photoreceptors.     

Evaluation of substrate promiscuity of an L‐carbamoyl amino acid amidohydrolase from Geobacillus stearothermophilus CECT43

N‐carbamoyl‐amino‐acid amidohydrolase (also known as N‐carbamoylase) is the stereospecific enzyme responsible for the chirality of the D ‐ or L ‐amino acid obtained in the “Hydantoinase Process.” This process is based on the dynamic kinetic resolution of D ,L ‐5‐monosubstituted hydantoins. In this work, we have demonstrated the capability of a recombinant L ‐N‐carbamoylase from the thermophilic bacterium Geobacillus stearothermophilus CECT43 (BsLcar) to hydrolyze N‐acetyl and N‐formyl‐L ‐amino acids as well as the known N‐carbamoyl‐L ‐amino acids, thus proving its substrate promiscuity. BsLcar showed faster hydrolysis for N‐formyl‐L ‐amino acids than for N‐carbamoyl and N‐acetyl‐L ‐derivatives, with a catalytic efficiency (k_cat/K_m) of 8.58 × 10⁵, 1.83 × 10⁴, and 1.78 × 10³ (s^?1 M^?1), respectively, for the three precursors of L ‐methionine. Optimum reaction conditions for BsLcar, using the three N‐substituted‐L ‐methionine substrates, were 65°C and pH 7.5. In all three cases, the metal ions Co²⁺, Mn²⁺, and Ni²⁺ greatly enhanced BsLcar activity, whereas metal‐chelating agents inhibited it, showing that BsLcar is a metalloenzyme. The Co²⁺‐dependent activity profile of the enzyme showed no detectable inhibition at high metal ion concentrations. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010     

The Safety and Efficacy of Antifibrinolytic Therapy in Neonatal Cardiac Surgery

BackgroundNeonates undergoing open-heart surgery are particularly at risk of postoperative bleeding requiring blood transfusion. Aprotinin has attained high efficacy in reducing the requirement for a blood transfusion following a cardiopulmonary bypass, but is seldom studied in the neonatal age group. The aim of this study was to compare the efficacy and adverse effects of aprotinin and tranexamic acid in neonates undergoing open-heart surgery at a single centre.MethodsBetween October 2003 and March 2008, perioperative data of 552 consecutive neonatal patients undergoing open-heart surgery in Children’s Hospital Boston were reviewed. Among them, 177 did not receive antifibrinolytic therapy (Group A); 100 were treated with tranexamic acid only (Group B); and 275 patients received aprotinin with or without tranexamic acid (Group C). Except for antifibrinolytic therapy, the anaesthesiological and surgical protocols remained identical. Postoperative complications and in-hospital mortality were the primary study endpoints.ResultsBody weight and Risk Adjustment for Congenital Heart Surgery (RACHS-1) scores were statistically comparable among the three groups. No statistically significant differences were observed between the duration of hospitalization, chest tube drainage, reexploration for bleeding, and kidney function impairment. In Group C, less blood was transfused within 24 hours than in GroupB. Operative mortality was similar among the three groups.ConclusionNo further risk and kidney injury were observed in the use of aprotinin in neonatal cardiac surgery, aprotinin demonstrated a reduced requirement for blood transfusion compared with tranexamic acid. Our data provide reasonable evidence that aprotinin and tranexamic acid are safe and efficacious as antifibrinolytic modalities in neonatal patients undergoing cardiac surgery.     

Spatial and temporal regulation of the metabolism of reactive oxygen and nitrogen species during the early development of pepper (Capsicum annuum) seedlings

Background and Aims The development of seedlings involves many morphological, physiological and biochemical processes, which are controlled by many factors. Some reactive oxygen and nitrogen species (ROS and RNS, respectively) are implicated as signal molecules in physiological and phytopathological processes. Pepper (Capsicum annuum) is a very important crop and the goal of this work was to provide a framework of the behaviour of the key elements in the metabolism of ROS and RNS in the main organs of pepper during its development.Methods The main seedling organs (roots, hypocotyls and green cotyledons) of pepper seedlings were analysed 7, 10 and 14 d after germination. Activity and gene expression of the main enzymatic antioxidants (catalase, ascorbate–glutathione cycle enzymes), NADP-generating dehydrogenases and S-nitrosoglutathione reductase were determined. Cellular distribution of nitric oxide (^·NO), superoxide radical (O₂^·–) and peroxynitrite (ONOO^–) was investigated using confocal laser scanning microscopy.Key Results The metabolism of ROS and RNS during pepper seedling development was highly regulated and showed significant plasticity, which was co-ordinated among the main seedling organs, resulting in correct development. Catalase showed higher activity in the aerial parts of the seedling (hypocotyls and green cotyledons) whereas roots of 7-d-old seedlings contained higher activity of the enzymatic components of the ascorbate glutathione cycle, NADP-isocitrate dehydrogenase and NADP-malic enzyme.Conclusions There is differential regulation of the metabolism of ROS, nitric oxide and NADP dehydrogenases in the different plant organs during seedling development in pepper in the absence of stress. The metabolism of ROS and RNS seems to contribute significantly to plant development since their components are involved directly or indirectly in many metabolic pathways. Thus, specific molecules such as H₂O₂ and NO have implications for signalling, and their temporal and spatial regulation contributes to the success of seedling establishment.