Lead accumulation in the aquatic fern Azolla filiculoides.

In this study, we characterized lead (Pb2+) accumulation and storage by the aquatic fern Azolla filiculoides. Lead precipitates were detected in the vacuoles of mesophyll cells of Azolla plants cultured for 6 d in rich growth medium containing 20 mg l(-1) Pb2+. Energy dispersive spectroscopy (EDS) analysis of the relative element content of leaves collected from these plants revealed a 100% increase in the levels of P, S, Na and Ca and a 40% decrease in Mg and Cl compared to the untreated plants. Both Azolla whole plants and isolated apoplasts were incubated for 6 d in 20 mg l(-1) Pb2+. Lead content in the whole plant composed 0.37%, 2.3% and 1.8% of the dry weight after 2, 4 and 6 d of growth, respectively, while the isolated Azolla apoplast contained 0.125%, 1.22% and 1.4% Pb2+, respectively. Lead content in Azolla whole plant increase by 200%, 100% and 22% after 2, 4 and 6 d of growth, respectively, when compared to Azolla apoplast. Dark, electron dense deposits of lead were observed in light and transmission electron microscope in leaf cells treated with lead. All the observed lead deposits were localized in vacuoles while larger lead deposits were found in mature leaves than in young leaves. No lead deposits were found in cells of the cyanobiont Anabaena when the plants were exposed to similar conditions. Activity and content of V-H+-ATPase were studied in Azolla plants grown in the presence of 20, 40 and 80 mg l(-1) of lead for a period of 4 d. Activity of V-H+-ATPase was increased by 190%, 210% and 220%, respectively, but the content of V-H+-ATPase was reduced by all lead concentrations.     

Large-Scale Preparation of Recombinant Ovine Prolactin and Determination of Its in Vitro and in Vivo Activity

Recombinant bovine Ala-prolactin (PRL) (GenBank Accession No. V00112) in prokaryotic expression plasmid pMON3401 was mutated using a mutagenesis kit, to prepare plasmid encoding ovine PRL (oPRL) (GenBank Accession No. M27057) Escherichia coli cells transformed with this latter plasmid overexpressed large amounts of oPRL upon induction with nalidixic acid. The expressed protein, found in inclusion bodies, was refolded and purified to homogeneity on a Q-Sepharose column, yielding an electrophoretically pure fraction composed of over 98% monomeric protein of the expected molecular mass of 23 kDa. The biological activity of the recombinant oPRL after proper renaturation was evidenced in vitro by its ability to stimulate proliferation of rat lymphoma Nb₂ cells possessing PRL receptors, to stimulate luciferase activity in HEK 293 cells transiently transfected with oPRL receptors, and to induce progesterone secretion in primary cultures of luteal cells obtained from midpregnant ewes. In contrast to ovine growth hormone or ovine placental lactogen, recombinant oPRL had no galactopoietic effect in lactating ewes.     

Breaking dormancy during flood and drought: sublethal growth and physiological responses of three emergent wetland herbs used in bioretention basins

Bioretention basins are man-made topographic depressions designed to collect and retain surface water runoff. In most cases these basins are used to prevent flooding and/or remove environmentally harmful pollutants and sediments from entering natural aquatic systems. To maximize environmental benefits, these systems are often planted with flood-tolerant wetland hydrophytes that are capable of withstanding extended periods of drought. Unfortunately, little is known about how these plants respond to extreme hydrologies while breaking seasonal dormancy. The purpose of this study was to better understand the morphological and physiological responses of three wetland species (Pontederia cordata L., Saururus cernuus L., and Schoenoplectus tabernaemontani C.C. Gmel. Palla) often used in bioretention basins while emerging from dormancy in either flooded or drought conditions. Results indicate that only S. tabernaemontani was affected morphologically by drought with lower leaf area and aboveground biomass. While significant reductions in stomatal surface indices were also observed in drought-treated S. tabernaemontani, all three species had reductions in stomatal conductance (g) when grown in drier soils. Moreover, drought conditions promoted decreases in leaf water potential (Ψ _leaf) for all three species, and reductions in tissue water content (θ) for P. cordata and S. tabernaemontani. Based on the overall morphological and physiological responses, S. cernuus maintained the lowest productivity, and appeared to be the best suited for tolerating sustained soil water deficits. If high plant productivity is desired, however, S. tabernaemontani was able to maintain high plant growth while making necessary modifications that facilitated greater drought-resistance.     

Woody vegetation composition and diversity in woodlands inside and outside a Forest Reserve in Jos,Nigeria

Protected areas such as forest reserves are often assumed to be best ways to conserve biodiversity and maintain intact ecosystems. We examined woody plant composition and diversity in the gallery forest and savannah woodland habitats of Amurum Forest Reserve and areas immediately surrounding it in Jos, Nigeria. A total of 100 10 × 10 m sample plots were established inside and outside the reserve. All woody plants ≥1 cm diameter at breast height (dbh) were identified and measured. A total of 7,564 individual plants categorized as 134 species from 44 families were recorded. Overall species diversity was significantly higher in the Forest Reserve than outside the reserve, although more species were encountered outside the reserve. Our findings suggest that, protected areas and the areas surrounding them are important for the conservation of biodiversity as the areas outside protected areas also contain species of conservation value. The continuous degrading areas outside protected areas isolates them and poses a serious threat to the long‐term viability of wildlife populations, so it is important that biodiversity in protected areas and their surrounding areas be conserved.     

Integrating Ethnography and Hunting Sustainability Modeling for Primate Conservation in an Indigenous Reserve in Guyana

Indigenous reserves are increasingly common throughout the tropical world. This is particularly true in Amazonia, where they make up >50% of protected land area. While these reserves offer tremendous opportunities for conservation, hunting represents a considerable threat to primate populations. As eliminating hunting may not be feasible, conservationists must work collaboratively with indigenous groups to promote sustainable management. This requires an understanding of the sociocultural drivers of hunting, quantitatively assessing sustainability, and developing co-management strategies that are commensurable with indigenous ontologies. In this article, we integrate ethnography with sustainability modeling to assess the importance of primate hunting to the livelihoods and culture of indigenous Waiwai in the Konashen Community-Owned Conservation Area, Guyana and to simultaneously promote sustainable co-management. We collected quantitative data on Waiwai harvesting through hunter self-monitoring and used semistructured interviews, unstructured interviews, and participant observation to understand the cultural importance of hunting to Waiwai society. We incorporated these data into spatially explicit biodemographic models to assess sustainability for four primate species. Primates, particularly spider monkeys (Ateles pansicus), were among the most important Waiwai prey and primate hunting played an important role in the construction of both individual and collective Waiwai identity. Our biodemographic models indicated that hunting will cause relatively little depletion for most primates in 20 years, although spider monkeys are predicted to disappear from a majority of the Waiwai catchment area. We argue that successful co-management of hunting in indigenous reserves requires truly integrative approaches that combine quantitative sustainability assessments with detailed concurrent ethnographic research.     

Characterization of the fluorescence of the protamine thynnine and studies of binding to double-stranded DNA

The protamine thynnine is an arginine-rich protein approximately 30 amino acids long with a tyrosine in the middle of its sequence. Its fluorescence decay kinetics can be described by a biexponential function with lifetimes of 0.52 and 2.1 ns, with almost equal preexponential factors. The fluorescence quencher CsCl does not affect the short lifetime but shifts the equilibrium between the long and short lifetime toward the short one and reduces the long lifetime. In nature, thynnine is found complexed with chromosomal DNA. In vitro complexes of thynnine with double-stranded (ds) DNA are stable at physiologic ionic strength but dissociate at high NaCl concentration. This dissociation can be monitored by steady-state fluorescence. From the salt concentration dependence of the dissociation of the complex of thynnine with ds-DNA 145 bp long, it can be concluded that only 4 of 21 possible full electrostatic bonds are involved in thynnine-DNA binding. In addition, the binding constant at 1M NaCl is of the order of 10⁶, indicating a strong nonelectrostatic component in arginine-DNA binding.     

MzrA: a novel modulator of the EnvZ/OmpR two-component regulon

Analysis of suppressors that alleviate the acute envelope stress phenotype of a Δ bamB Δ degP strain of Escherichia coli identified a novel protein MzrA and pleiotropic envZ mutations. Genetic evidence shows that overexpression of MzrA – formerly known as YqjB and EcfM – modulates the activity of EnvZ/OmpR similarly to pleiotropic EnvZ mutants and alter porin expression. However, porin expression in strains devoid of MzrA or overexpressing it is still sensitive to medium osmolarity, pH and procaine, all of which modulate EnvZ/OmpR activities. Thus, MzrA appears to alter the output of the EnvZ/OmpR system but not its ability to receive and respond to various environmental signals. Localization and topology experiments indicate that MzrA is a type II membrane protein, with its N-terminus exposed in the cytoplasm and C-terminus in the periplasm. Bacterial two-hybrid experiments determined that MzrA specifically interacts with EnvZ but not with OmpR or the related membrane sensor kinase, CpxA. This and additional genetic and biochemical evidence suggest that the interaction of MzrA with EnvZ would either enhance EnvZ's kinase activity or reduce its phosphatase activity, thus elevating the steady state levels of OmpR∼P. Furthermore, our data show that MzrA links the two-component envelope stress response regulators, CpxA/CpxR and EnvZ/OmpR.