An Age-Wise Comparison of Human Airway Smooth Muscle Proliferative Capacity

We compared the proliferation of neonatal and adult airway smooth muscle cells (ASMC) with no/moderate lung disease, in glucose- (energy production by glycolysis) or glucose-free medium (ATP production from mitochondrial oxidative phosphorylations only), in response to 10% fetal calf serum (FCS) and PDGF-AA. In the presence of glucose, cell counts were significantly greater in neonatal vs. adult ASMC. Similarly, neonatal ASMC DNA synthesis in 10% FCS and PDGF-AA, and [Ca²⁺]i responses in the presence of histamine were significantly enhanced vs. adults. In glucose-free medium, cell proliferation was preserved in neonatal cells, unlike in adult cells, with concomitant increased porin (an indicator of mitochondrial activity) protein expression. Compared to adults, stimulated neonatal human ASMC are in a rapid and robust proliferative phase and have the capacity to respond disproportionately under abnormal environmental conditions, through increased mitochondrial biogenesis and altered calcium homeostasis.     

Fungal Bioweathering of Mimetite and a General Geomycological Model for Lead Apatite Mineral Biotransformations

Fungi play important roles in biogeochemical processes such as organic matter decomposition, bioweathering of minerals and rocks, and metal transformations and therefore influence elemental cycles for essential and potentially toxic elements, e.g., P, S, Pb, and As. Arsenic is a potentially toxic metalloid for most organisms and naturally occurs in trace quantities in soil, rocks, water, air, and living organisms. Among more than 300 arsenic minerals occurring in nature, mimetite [Pb₅(AsO₄)₃Cl] is the most stable lead arsenate and holds considerable promise in metal stabilization for in situ and ex situ sequestration and remediation through precipitation, as do other insoluble lead apatites, such as pyromorphite [Pb₅(PO₄)₃Cl] and vanadinite [Pb₅(VO₄)₃Cl]. Despite the insolubility of mimetite, the organic acid-producing soil fungus Aspergillus niger was able to solubilize mimetite with simultaneous precipitation of lead oxalate as a new mycogenic biomineral. Since fungal biotransformation of both pyromorphite and vanadinite has been previously documented, a new biogeochemical model for the biogenic transformation of lead apatites (mimetite, pyromorphite, and vanadinite) by fungi is hypothesized in this study by application of geochemical modeling together with experimental data. The models closely agreed with experimental data and provided accurate simulation of As and Pb complexation and biomineral formation dependent on, e.g., pH, cation-anion composition, and concentration. A general pattern for fungal biotransformation of lead apatite minerals is proposed, proving new understanding of ecological implications of the biogeochemical cycling of component elements as well as industrial applications in metal stabilization, bioremediation, and biorecovery.     

Isolation,Culture and Long-Term Maintenance of Primary Mesencephalic Dopaminergic Neurons From Embryonic Rodent Brains

Degeneration of mesencephalic dopaminergic (mesDA) neurons is the pathological hallmark of Parkinson’s diseae. Study of the biological processes involved in physiological functions and vulnerability and death of these neurons is imparative to understanding the underlying causes and unraveling the cure for this common neurodegenerative disorder. Primary cultures of mesDA neurons provide a tool for investigation of the molecular, biochemical and electrophysiological properties, in order to understand the development, long-term survival and degeneration of these neurons during the course of disease. Here we present a detailed method for the isolation, culturing and maintenance of midbrain dopaminergic neurons from E12.5 mouse (or E14.5 rat) embryos. Optimized cell culture conditions in this protocol result in presence of axonal and dendritic projections, synaptic connections and other neuronal morphological properties, which make the cultures suitable for study of the physiological, cell biological and molecular characteristics of this neuronal population.     

Photosynthetic Light Responses May Explain Vertical Distribution of Hymenophyllaceae Species in a Temperate Rainforest of Southern Chile

Some epiphytic Hymenophyllaceae are restricted to lower parts of the host (<60 cm; 10–100 μmol photons m^-2 s^-1) in a secondary forest of Southern Chile; other species occupy the whole host height (≥10 m; max PPFD >1000 μmol photons m^-2 s^-1). Our aim was to study the photosynthetic light responses of two Hymenophyllaceae species in relation to their contrasting distribution. We determined light tolerance of Hymenoglossum cruentum and Hymenophyllum dentatum by measuring gas exchange, PSI and PSII light energy partitioning, NPQ components, and pigment contents. H. dentatum showed lower maximum photosynthesis rates (A_max) than H. cruentum, but the former species kept its net rates (A_n) near A_max across a wide light range. In contrast, in the latter one, A_n declined at PPFDs >60 μmol photons m^-2 s^-1. H. cruentum, the shadiest plant, showed higher chlorophyll contents than H. dentatum. Differences in energy partitioning at PSI and PSII were consistent with gas exchange results. H. dentatum exhibited a higher light compensation point of the partitioning of absorbed energy between photochemical Y(PSII) and non-photochemical Y(NPQ) processes. Hence, both species allocated energy mainly toward photochemistry instead of heat dissipation at their light saturation points. Above saturation, H. cruentum had higher heat dissipation than H. dentatum. PSI yield (YPSI) remained higher in H. dentatum than H. cruentum in a wider light range. In both species, the main cause of heat dissipation at PSI was a donor side limitation. An early dynamic photo-inhibition of PSII may have caused an over reduction of the Qa⁺ pool decreasing the efficiency of electron donation to PSI. In H. dentatum, a slight increase in heat dissipation due to acceptor side limitation of PSI was observed above 300 μmol photons m^-2s^-1. Differences in photosynthetic responses to light suggest that light tolerance and species plasticity could explain their contrasting vertical distribution.     

Reproductive characteristics that favor invasiveness in Leonotis nepetifolia (L.) R. Br.

Reproductive systems are life attributes important in defining the demography and genetic constitution of invasive alien species populations. We describe the phenology, floral behavior and floral visitors in Mexican populations of Leonotis nepetifolia considered invasive in America, Asia and Oceania. The mating system was determined through pollination experiments and, with a morphological analysis of flowers (outcrossing index, OCI) and pollen/ovule ratio, the breeding system was evaluated. Germination of 1 and 2-year-old seeds was tested to assess the potential characteristics of germination. Leonotis nepetifolia was reproductive for 7 months (June to December) and tended towards a specific season during autumn. Anthesis lasted 36 hr with protogyny and no hercogamy, with floral visitors of Apodiformes, Hymenoptera, Lepidoptera and Thysanoptera. Pollination experiments indicated a mixed mating system, whereas the OCI and the pollen/ovule ratio pointed towards a facultative xenogamous breeding system. Seed production was high (1,445 ± 132 seeds/plant); the seeds had potential longevity and were neutral photoblastic. One-year-old seeds germinated slightly later ( = 2.6 ± 0.11 days) than 2-year-old seeds ( = 1.9 ± 0.02 days), both synchronously (IS_1yr = 0.88 ± 0.03 and IS_2yr = 0.82 ± 0.02). Germination percentage for 1-year-old seeds was lower (55.33 ± 4.40%) than that of 2-year-old seeds (94.18 ± 0.59%), suggesting a potential longevity of the seeds in an optimal environment. Reproductive characteristics, such as wide reproductive period, mixed breeding system, copious seed production, seeds with potential longevity, and quick and synchronic germination in different light conditions, favor the invasive capacity of Leonotis nepetifolia.     

Diversity of spiders and orthopterans respond to intra-seasonal and spatial environmental changes

Our understanding of arthropod responses to environmental pressures is limited, especially for the poorly studied Mediterranean region. In the light of likely further environmental change and the need for protocols for rapid biodiversity assessment, we measured how the abundance and species richness of two taxa, ground spiders and Orthoptera, belonging to different functional groups, fluctuates intra- seasonally (early-mid-late summer) and across habitat types (grasslands, maquis, forests). We also tested their surrogate value. Spiders were found to have higher species richness and abundance almost throughout the investigation. Orthoptera had lower species richness and abundance in forests compared to grasslands and maquis, while no significant difference between habitats was revealed for spiders. Early-summer was the richest period for spiders while mid-summer was the richest for Orthoptera. Canopy cover was found to significantly influence community composition of both groups, while herb height and cover of stones was a determinant factor for Orthoptera only. There was a significant congruence between the two groups and Orthoptera provided the best complementary network. Our results show that diversity patterns of both spiders and Orthoptera are sensitive to environmental changes even over short time-scales (e.g. within the summer period) and space (e.g. across different habitat types), suggesting that small inexpensive experimental designs may still reveal community dynamics. For conservation purposes, we advise a focus on variables regulating habitat heterogeneity and microhabitat characteristics. We provide a list of the most influential species and propose the most effective network for obtaining information on the local fauna.