Purinergic and nitrergic junction potential in the human colon

The aim of the present work is to investigate a putative junction transmission [nitric oxide (NO) and ATP] in the human colon and to characterize the electrophysiological and mechanical responses that might explain different functions from both neurotransmitters. Muscle bath and microelectrode techniques were performed on human colonic circular muscle strips. The NO donor sodium nitroprusside (10 microM), but not the P2Y receptor agonist adenosine 5'-O-2-thiodiphosphate (10 microM), was able to cause a sustained relaxation. NG-nitro-L-arginine (L-NNA) (1 mM), a NO synthase inhibitor, but not 2'-deoxy-N6-methyl adenosine 3',5'-diphosphate tetraammonium salt (MRS 2179) (10 microM), a P2Y antagonist, increased spontaneous motility. Electrical field stimulation (EFS) at 1 Hz caused fast inhibitory junction potentials (fIJPs) and a relaxation sensitive to MRS 2179 (10 microM). EFS at higher frequencies (5 Hz) showed biphasic IJP with fast hyperpolarization sensitive to MRS 2179 followed by sustained hyperpolarization sensitive to L-NNA; both drugs were needed to fully block the EFS relaxation at 2 and 5 Hz. Two consecutive single pulses induced MRS 2179-sensitive fIJPs that showed a rundown. The rundown mechanism was not dependent on the degree of hyperpolarization and was present after incubation with L-NNA (1 mM), hexamethonium (100 microM), MRS 2179 (1 microM), and NF023 (10 microM). We concluded that single pulses elicit ATP release from enteric motor neurons that cause a fIJP and a transient relaxation that is difficult to maintain over time; also, NO is released at higher frequencies causing a sustained hyperpolarization and relaxation. These differences might be responsible for complementary mechanisms of relaxation being phasic (ATP) and tonic (NO).     

Micropatterning of single endothelial cell shape reveals a tight coupling between nuclear volume in G1 and proliferation

Shape-dependent local differentials in cell proliferation are considered to be a major driving mechanism of structuring processes in vivo, such as embryogenesis, wound healing, and angiogenesis. However, the specific biophysical signaling by which changes in cell shape contribute to cell cycle regulation remains poorly understood. Here, we describe our study of the roles of nuclear volume and cytoskeletal mechanics in mediating shape control of proliferation in single endothelial cells. Micropatterned adhesive islands were used to independently control cell spreading and elongation. We show that, irrespective of elongation, nuclear volume and apparent chromatin decondensation of cells in G1 systematically increased with cell spreading and highly correlated with DNA synthesis (percent of cells in the S phase). In contrast, cell elongation dramatically affected the organization of the actin cytoskeleton, markedly reduced both cytoskeletal stiffness (measured dorsally with atomic force microscopy) and contractility (measured ventrally with traction microscopy), and increased mechanical anisotropy, without affecting either DNA synthesis or nuclear volume. Our results reveal that the nuclear volume in G1 is predictive of the proliferative status of single endothelial cells within a population, whereas cell stiffness and contractility are not. These findings show that the effects of cell mechanics in shape control of proliferation are far more complex than a linear or straightforward relationship. Our data are consistent with a mechanism by which spreading of cells in G1 partially enhances proliferation by inducing nuclear swelling and decreasing chromatin condensation, thereby rendering DNA more accessible to the replication machinery.     

Ultrastructure of Early Secondary Embryogenesis by Multicellular and Unicellular Pathways in Cork Oak (Quercus suber L.)

Early cellular events during secondary embryogenesis were studiedin a cork oak recurrent embryogenic system in which embryosarise either in a multicellular budding pathway from a compactmass of proliferation or from isolated single cells in friablecallus. The compact mass of proliferation originated from theepidermal cells at the hypocotyl whose growth and convolutionwas characterized by a decrease in the nucleus/cytoplasm ratioand a marked increase in storage products. The transition fromthe compact mass to meristematic primordia occurred at the peripheryand was accompanied by cell dedifferentiation and a drasticreduction of storage products. Meristematic primordia evolvedto globular embryos by the organization of a protodermis andtwo internal centres. Microscope analysis of friable callusshowed an hypothetical sequence from single cells to aggregatesof a few cells, meristematic cell clusters and globular embryos.Single cells showed typical features of embryogenic cells suchas rich cytoplasm and a large number of starch grains and lipidbodies. A progressive cell dedifferentiation and a drastic reductionof storage products was observed when aggregates of a few cellsand meristematic cell clusters were compared. Progressive bipolarizationin large meristematic cell clusters initiated globular embryoformation. The comparison of both embryogenic pathways at theultrastructural level showed that subcellular changes followa similar sequential pattern, especially with regard to thestorage products. The possible role of plastid extrusions andmultivesicular bodies in the changing pattern of starch metabolismduring embryogenesis is discussed. Copyright 2001 Annals ofBotany Company Quercus suber L, cork oak, somatic embryogenesis, multicellular budding, friable callus, ultrastructural studies     

From Life Cycle Assessment to Life Cycle Management

Life cycle assessment (LCA) is a widely accepted methodology to support decision‐making processes in which one compares alternatives, and that helps prevent shifting of environmental burdens along the value chain or among impact categories. According to regulation in the European Union (EU), the movement of waste needs to be reduced and, if unavoidable, the environmental gain from a specific waste treatment option requiring transport must be larger than the losses arising from transport. The EU explicitly recommends the use of LCA or life cycle thinking for the formulation of new waste management plans. In the last two revisions of the Industrial Waste Management Programme of Catalonia (PROGRIC), the use of a life cycle thinking approach to waste policy was mandated. In this article we explain the process developed to arrive at practical life cycle management (LCM) from what started as an LCA project. LCM principles we have labeled the “3/3” principle or the “good enough is best” principle were found to be essential to obtain simplified models that are easy to understand for legislators and industries, useful in waste management regulation, and, ultimately, feasible. In this article, we present the four models of options for the management of waste solvent to be addressed under Catalan industrial waste management regulation. All involved actors concluded that the models are sufficiently robust, are easy to apply, and accomplish the aim of limiting the transport of waste outside Catalonia, according to the principles of proximity and sufficiency.     

Wnt-5a Ligand Modulates Mitochondrial Fission-Fusion in Rat Hippocampal Neurons

The Wnt signaling pathway plays an important role in developmental processes, including embryonic patterning, cell specification, and cell polarity. Wnt components participate in the development of the central nervous system, and growing evidence indicates that this pathway also regulates the function of the adult nervous system. In this study, we report that Wnt-5a, a noncanonical Wnt ligand, is a potent activator of mitochondrial dynamics and induces acute fission and fusion events in the mitochondria of rat hippocampal neurons. The effect of Wnt-5a was inhibited in the presence of sFRP, a Wnt scavenger. Similarly, the canonical Wnt-3a ligand had no effect on mitochondrial fission-fusion events, suggesting that this effect is specific for Wnt-5a alone. We also show that the Wnt-5a effects on mitochondrial dynamics occur with an increase in both intracellular and mitochondrial calcium (Ca²⁺), which was correlated with an increased phosphorylation of Drp1(Ser-616) and a decrease of Ser-637 phosphorylation, both indicators of mitochondrial dynamics. Electron microscope analysis of hippocampal tissues in the CA1 region showed an increase in the number of mitochondria present in the postsynaptic region, and this finding correlated with a change in mitochondrial morphology. We conclude that Wnt-5a/Ca²⁺ signaling regulates the mitochondrial fission-fusion process in hippocampal neurons, a feature that might help to further understand the role of Wnt-related pathologies, including neurodegenerative diseases associated with mitochondrial dysfunction, and represents a potentially important link between impaired metabolic function and degenerative disorders.     

Alien plants of coastal dune habitats in southern Spain

Alien plants from psammophilous ecosystems have been studied in southern Spain. Sixteen sites, which are included in the Andalusian Network of Protected Spaces, have been chosen along 1100 km of Andalusian coasts in order to verify the degree of abundance and influence of these species on different communities. Out of the 26 localized species, the increase of some invasive plants, such as Arctotheca calendula, Agave americana, A. sisalana, Cortaderia selloana, Lantana camara and Opuntia dillenii, affects the conservation of some natural coastal ecosystems.     

The phlebotomine fauna (Diptera: Psychodidae) of Guaraí, state of Tocantins,with an emphasis on the putative vectors of American cutaneous leishmaniasis in rural settlement and periurban areas

Phlebotomine sandflies were captured in rural settlement and periurban areas of the municipality of Guaraí in the state of Tocantins (TO), an endemic area of American cutaneous leishmaniasis (ACL). Forty-three phlebotomine species were identified, nine of which have already been recognised as ACL vectors. Eleven species were recorded for the first time in TO. Nyssomyia whitmani was the most abundant species, followed by Evandromyia bourrouli, Nyssomyia antunesi and Psychodopygus complexus. The Shannon-Wiener diversity index and the evenness index were higher in the rural settlement area than in the periurban area. The evaluation of different ecotopes within the rural area showed the highest frequencies of Ev. bourrouli and Ny. antunesi in chicken coops, whereas Ny. whitmani predominated in this ecotope in the periurban area. In the rural settlement area, Ev. bourrouli was the most frequently captured species in automatic light traps and Ps. complexus was the most prevalent in Shannon trap captures. The rural settlement environment exhibited greater phlebotomine biodiversity than the periurban area. Ps. complexus and Psychodopygus ayrozai naturally infected with Leishmania (Viannia) braziliensis were identified. The data identified Ny. whitmani as a potential ACL vector in the periurban area, whereas Ps. complexus was more prevalent in the rural environment associated with settlements.