Root Growth and Oxygen Relations at Low Water Potentials. Impact of Oxygen Availability in Polyethylene Glycol Solutions1

Polyethylene glycol (PEG), which is often used to impose low water potentials (ψw) in solution culture, decreases O2 movement by increasing solution viscosity. We investigated whether this property causes O2 deficiency that affects the elongation or metabolism of maize (Zea mays L.) primary roots. Seedlings grown in vigorously aerated PEG solutions at ambient solution O2 partial pressure (pO2) had decreased steady-state root elongation rates, increased root-tip alanine concentrations, and decreased root-tip proline concentrations relative to seedlings grown in PEG solutions of above-ambient pO2 (alanine and proline accumulation are responses to hypoxia and low ψw, respectively). Measurements of root pO2 were made using an O2 microsensor to ensure that increased solution pO2 did not increase root pO2 above physiological levels. In oxygenated PEG solutions that gave maximal root elongation rates, root pO2 was similar to or less than (depending on depth in the tissue) pO2 of roots growing in vermiculite at the same ψw. Even without PEG, high solution pO2 was necessary to raise root pO2 to the levels found in vermiculite-grown roots. Vermiculite was used for comparison because it has large air spaces that allow free movement of O2 to the root surface. The results show that supplemental oxygenation is required to avoid hypoxia in PEG solutions. Also, the data suggest that the O2 demand of the root elongation zone may be greater at low relative to high ψw, compounding the effect of PEG on O2 supply. Under O2-sufficient conditions root elongation was substantially less sensitive to the low ψw imposed by PEG than that imposed by dry vermiculite.

Differential toxicity of mitomycin C and porfiromycin to aerobic and hypoxic Chinese hamster ovary cells overexpressing human NADPH:cytochrome c (P-450) reductase.

Purified NADPH:cytochrome c (P-450) reductase (FpT; NADPH-ferrihemoprotein oxidoreductase, EC 1.6.2.4) can reductively activate mitomycin antibiotics through a one-electron reduction to species that alkylate DNA. To assess the involvement of FpT in the intracellular activation of the mitomycins, transfectants overexpressing a human FpT cDNA were established from a Chinese hamster ovary cell line deficient in dihydrofolate reductase (CHO-K1/dhfr-). The parental cell line was equisensitive to the cytotoxic action of mitomycin C under oxygenated and hypoxic conditions. In contrast, porfiromycin was considerably less cytotoxic to wild-type parental cells than was mitomycin C in air and markedly more cytotoxic under hypoxia. Two FpT-transfected clones were selected that expressed 19- and 27-fold more FpT activity than the parental line. Levels of other oxidoreductases implicated in the activation of the mitomycins were unchanged. Significant increases in sensitivity to mitomycin C and porfiromycin in the two FpT-transfected clones were seen under both oxygenated and hypoxic conditions, with the increases in toxicity being greater under hypoxia than in air. These findings demonstrate that FpT can bioreductively activate the mitomycins in living cells and implicate FpT in the differential aerobic/hypoxic toxicity of the mitomycins.

Lipoxygenase-catalyzed oxygenation of storage lipids is implicated in lipid mobilization during germination.

The etiolated germination process of oilseed plants is characterized by the mobilization of storage lipids, which serve as a major carbon source for the seedling. We found that during early stages of germination in cucumber, a lipoxygenase (linoleate: oxygen oxidoreductase, EC 1.13.11.12) form is induced that is capable of oxygenating the esterified fatty acids located in the lipid-storage organelles, the so-called lipid bodies. Large amounts of esterified (13S)-hydroxy-(9Z,11E)-octadecadienoic acid were detected in the lipid bodies, whereas only traces of other oxygenated fatty acid isomers were found. This specific product pattern confirms the in vivo action of this lipoxygenase form during germination. Lipid fractionation studies of lipid bodies indicated the presence of lipoxygenase products both in the storage triacylglycerols and, to a higher extent, in the phospholipids surrounding the lipid stores as a monolayer. The degree of oxygenation of the storage lipids increased drastically during the time course of germination. We show that oxygenated fatty acids are preferentially cleaved from the lipid bodies and are subsequently released into the cytoplasm. We suggest that they may serve as substrate for beta-oxidation. These data suggest that during the etiolated germination, a lipoxygenase initiates the mobilization of storage lipids. The possible mechanisms of this implication are discussed.

Ergosteroids: induction of thermogenic enzymes in liver of rats treated with steroids derived from dehydroepiandrosterone.

Dehydroepiandrosterone (DHEA), an intermediate in the biosynthesis of testosterone and estrogens, exerts several physiological effects not involving the sex hormones. When fed to rats it induces the thermogenic enzymes mitochondrial sn-glycerol-3-phosphate dehydrogenase and cytosolic malic enzyme in their livers. Animals and humans, and their excised tissues, are known to hydroxylate DHEA at several positions and to interconvert 7 alpha-hydroxy-DHEA, 7 beta-hydroxy-DHEA, 7-oxo-DHEA, and the corresponding derivatives of androst-5-enediol. We report here that these 7-oxygenated derivatives are active inducers of these thermogenic enzymes in rats and that the 7-oxo derivatives are more active than the parent steroids. We postulate that the 7 alpha-hydroxy and 7-oxo derivatives are on a metabolic pathway from DHEA to more active steroid hormones. These 7-oxo steroids have potential as therapeutic agents because of their increased activity and because they are not convertible to either testosterone or estrogens.

Estudo limnológico na represa Carlos Botelho (Itirapina/Brotas-SP): uma reavaliação comparativa do sistema Lobo-Broa

Os reservatórios, por serem corpos de água dinâmicos e pelos seus usos múltiplos, sofrem acelerado processo de eutrofização. Neste estudo avaliou-se a atual condição da represa do Lobo (municípios de Brotas e Itirapina, SP) em relação a algumas características físicas, químicas e biológicas em uma perspectiva temporal. As amostragens foram realizadas em duas épocas representativas das estações seca e chuvosa, compreendidas no período de setembro de 2000 e junho de 2001. Foram estabelecidos seis pontos de coleta, nos quais foram coletadas amostras de água para análises de nutrientes, clorofila, material em suspensão, zooplâncton e foram também realizadas medidas \"in situ\" de variáveis físicas e químicas, com o multisensor Horiba U10. Os resultados obtidos revelaram que ocorreram variações sazonais de moderada amplitude, caracterizando dois períodos: o seco com temperaturas menores, águas mais oxigenadas, menor condutividade elétrica e maiores concentrações de nutrientes. O zooplâncton da represa do Lobo foi numericamente dominado pelos rotíferos, seguidos pelos cladóceros e copépodos em proporções bem menores, similarmente ao reportado pelos estudos anteriores, indicando que em nível de grandes grupos não houve ainda uma mudança na estrutura desta comunidade. As variações na densidade dos principais grupos zooplanctônicos estão relacionadas às características físicas e químicas das diferentes porções da represa (com maiores densidades na porção superior, mais eutrófica) e à sazonalidade, com maior abundância de rotíferos no período chuvoso e maiores densidades de protozoários e microcrustáceos no inverno. Durante o período de estudo a condição trófica do reservatório foi mesotrófica, contudo com níveis de nutrientes e biomassa do fitoplâncton, indicada pela concentração de clorofila, superiores aos níveis reportados para a década de 1970. Concluiu-se portanto que a represa está sofrendo um processo de eutrofização.

Contribuição ao conhecimento da malacofauna das camadas basais da Formação Corumbataí (Permiano), Estado de São Paulo

Os principais objetivos deste trabalho são: 1) estudar o material malacofaunístico de novos jazigos da região de Rio Claro - Piracicaba (SP) e de testemunhos de sondagem de Conchas (SP), referentes à porção basal da Formação Corumbataí, termo superior do Grupo Passa Dois de idade permiana e 2) determinar sua distribuição bioestratigráfica. Incluiu-se um exame parcial da provável correlação entre as faunas das bases das Formações Corumbataí (São Paulo) e Estrada Nova (Paraná) e considerações de ordem paleoecológica da malacofauna. A análise taxonômica dos bivalves da porção basal da Formação Corumbataí no Estado, forneceu apreciável diversidade genérica e específica. Duas secções colunares, uma de superfície e outra de subsuperfície são apresentadas, mostrando a distribuição bioestratigráfica dessa malacofauna. Os bivalves identificados e descritos nos diversos afloramentos foram: Angatubia cf. A. cowperesioides Mendes, Anthraconaia ? mezzalirai sp. n., Barbosaia cf. B. angulata Mendes, Barbosaia roxoi sp n., Casterella cf. C. camargoi Beurlen, Ferrazia simplicicarinata Mezzalira, Ferrazia sp., Holdhausiella elongata Mendes, Holdhausiella sp., Kidodia cf. K. stockleyi Cox, Mendesia piracicabensis gen. et sp. n., Plesiocyprinella sp., Rioclaroa cf. R. lefrevei Mezzalira. Com base na composição observada, sugeriu-se reunir as Zonas Leinzia froezi Mendes e Barbosaia angulata Mendes propostas por MEZZALIRA (1980) em uma única designada Zona Leinzia froezi - Barbosaia angulata. Os quatro níveis fossilíferos reconhecidos em subsuperfície (Poço 1-IG-Conchas) com as espécies: Casterella cf. C. gratiosa, Ferrazia cardinalis Reed e Pinzonella cf. P. illusa, a julgar pela distribuição vertical em relação à base da formação, situar-se-iam na Zona III de MEZZALIRA (1980) considerada quando definida como afossilífera. Dentre as principais conclusões, notou-se a predominância de bivalves com carenas simples e a duplas, provavelmente caráter adaptativo ao ambiente que seria provavelmente circunscrito, mixo-halino, de águas rasas, turvas e mal oxigenadas em certos intervalos.

Espécies polinucleares de cobre e ferro com catalisadores de oxidação e modelos de sítios ativos

Diferentes complexos de cobre(II), contendo ligantes do tipo base de Schiff e um grupamento imidazólico, com interesse bioinorgânico, catalítico e como novos materiais, foram preparados na forma de sais perclorato, nitrato ou cloreto e caracterizados através de diferentes técnicas espectroscópicas (UV/Vis, IR, EPR, Raman) e espectrometria de massa Tandem (ESI-MS/MS), além de análise elementar, condutividade molar e medidas de propriedades magnéticas. Alguns destes compostos, obtidos como cristais adequados, tiveram suas estruturas determinadas por cristalografia de raios-X. As espécies di- e polinucleares contendo pontes cloreto, mostraram desdobramentos das hiperfinas nos espectros de EPR, relacionados à presença do equilíbrio com a respectiva espécie mononuclear, devido à labilidade dos íons cloretos, dependendo do contra-íon e do tipo de solvente utilizado. Adicionalmente, em solução alcalina, estes compostos estão em equilíbrio com as correspondentes espécies polinucleares, onde os centros de cobre estão ligados através de um ligante imidazolato. Em meio alcalino, estes compostos polinucleares contendo ponte imidazolato foram também isolados e caracterizados por diferentes técnicas espectroscópicas e magnéticas. Através da variação estrutural e também do ligante-ponte foi possível modular o fenômeno da interação magnética entre os íons de cobre em estruturas correlatas di- e polinucleares. Os respectivos parâmetros magnéticos foram obtidos com ajuste das curvas experimentais de XM vs T, correlacionando-se muito bem com a geometria, ângulos e distâncias de ligação entre os íons, quando comparado com outros complexos similares descritos na literatura. Posteriormente, estudaram-se os fatores relacionados com a reatividade de todas essas espécies como catalisadores na oxidação de substratos de interesse (fenóis e aminas), através da variação do tamanho da cavidade nas estruturas cíclicas ou de variações no ligante coordenado ao redor do íon metálico. Vários deles se mostraram bons miméticos de tirosinases e catecol oxidases. Um novo complexo-modelo da citocromo c oxidase (CcO), utilizando a protoporfirina IX condensada ao quelato N,N,-bis[2-(1,2-metilbenzimidazolil)etil]amino e ao resíduo de glicil-L-histidina, foi sintetizado e caracterizado através de diferentes técnicas espectroscópicas, especialmente EPR. A adição de H2O2 ao sistema completamente oxidado, FeIII/CuII, a -55°C, ou o borbulhamento de oxigênio molecular a uma solução do complexo na sua forma reduzida, FeII/CuI, saturada de CO, resultou na formação de adutos com O2, de baixo spin, estáveis a baixas temperaturas.

Biodiesel wastes: An abundant and promising source for the preparation of acidic catalysts for utilization in etherification reaction

Environmentally friendly sulfonated black carbon (BC) catalysts were prepared from biodiesel waste, glycerol. These black carbons (BCs) contain a high amount of acidic groups, mainly sulfonated and oxygenated groups. Furthermore, these catalysts show a high catalytic activity in the glycerol etherification reaction with tert-butyl alcohol, the activity being larger for the sample prepared with a higher glycerol:sulfuric acid ratio (1:3). The yield for mono-tert-butyl glycerol (MTBG), di-tert-butyl glycerol (DTBG) and tri-tert-butyl-glycerol (TTBG) were very similar to those obtained using a commercial resin, Amberlyst-15. Furthermore, experimental results show that the carbon with the lowest acidic surface group content, BC prepared in minor glycerol:sulfuric acid ratio (10:1), can be chemically treated after carbonization to achieve an improved catalytic activity. The activity of all BCs is high and very similar, about 50% and 20% for the MTBG and DTBG + TTBG, respectively.

Evolution of the last koninckinids (Athyridida, Koninckinidae), a precursor signal of the early Toarcian mass extinction event in the Western Tethys

Koninckinids are a suitable group to shed light on the biotic crisis suffered by brachiopod fauna in the Early Jurassic. Koninckinid fauna recorded in the late Pliensbachian–early Toarcian from the easternmost Subbetic basin is analyzed and identified as a precursor signal for one of the most conspicuous mass extinction events of the Phylum Brachiopoda, a multi-phased interval with episodes of changing environmental conditions, whose onset can be detected from the Elisa–Mirabile subzones up to the early Toarcian extinction boundary in the lowermost Serpentinum Zone (T-OAE). The koninckinid fauna had a previously well-established migration pattern from the intra-Tethyan to the NW-European basins but a first phase with a progressive warming episode in the Pliensbachian–Toarcian transition triggered a koninckinid fauna exodus from the eastern/central Tethys toward the westernmost Mediterranean margins. A second stage shows an adaptive response to more adverse conditions in the westernmost Tethyan margins and finally, an escape and extinction phase is detected in the Atlantic areas from the mid-Polymorphum Zone onwards up to their global extinction in the lowermost Serpentinum Zone. This migration pattern is independent of the paleogeographic bioprovinciality and is unrelated to a facies-controlled pattern. The anoxic/suboxic environmental conditions should only be considered as a minor factor of partial control since well-oxygenated habitats are noted in the intra-Tethyan basins and this factor is noticeable only in the second westward migratory stage (with dwarf taxa and oligotypical assemblages). The analysis of cold-seep proxies in the Subbetic deposits suggests a radiation that is independent of methane releases in the Subbetic basin.

Novel chromone and xanthone derivatives: Synthesis and ROS/RNS scavenging activities

Chromones and xanthones are oxygen-containing heterocyclic compounds acknowledged by their antioxidant properties. In an effort to develop novel agents with improved activity, a series of compounds belonging to these chemical classes were prepared. Their syntheses involve the condensation of appropriate 2-methyl-4H-chromen-4-ones, obtained via Baker-Venkataraman rearrangement, with (E)-3-(3,4-dimethoxyphenyl)acrylaldehyde to provide the corresponding 2-[(1E,3E)-4-(3,4-dimethoxyphenyl)buta-1,3-dien-1-yl]-4H-chromen-4-ones. Subsequent electrocyclization and oxidation of these compounds led to the synthesis of 1-aryl-9H-xanthen-9-ones. After cleavage of the protecting groups, hydroxylated chromones and xanthones were assessed as scavenging agents against both reactive oxygen species (ROS) [superoxide radical (O2(•-)), hydrogen peroxide (H2O2), hypochlorous acid (HOCl), singlet oxygen ((1)O2), and peroxyl radical (ROO(•))] and reactive nitrogen species (RNS) [nitric oxide ((•)NO) and peroxynitrite anion (ONOO(-))]. Generally, all the tested new hydroxylated chromones and xanthones exhibited scavenger effects dependent on the concentration, with IC50 values found in the micromolar range. Some of them were shown to have improved scavenging activity when compared with previously reported analogues, allowing the inference of preliminary conclusions on the structure-activity relationship.

Synthesis of xanthone-1,2,3-triazole dyads

Xanthones and 1,2,3-triazoles are known to exhibit several biological, pharmacological and biocidal properties[1]. The potential applications of these two classes of heterocycles led us to develop new strategies to synthesize xanthone-1,2,3-triazole dyads, aiming to get potentially improved therapeutic agents[2]. With this rational in mind we designed and synthesized novel chromone derivatives 1a-d to be used as building motifs and to explore the reactivity of the two unsaturated systems (the diene and the alkyne). In the present communication we will present a new synthetic route towards the synthesis of xanthone-1,2,3-triazole dyads 7a-d using consecutively the azide-alkyne Huisgen 1,3-dipolar cycloaddition and Diels-Alder reaction. Our approach involves the synthesis chromone-triazole derivatives 2a-d using the reaction of 1a-d with sodium azide, followed by the methylation of the NH of the triazole moiety. The methylation afforded three isomers 3a-d, 4a-d and 5a-d, as expected. The major isomers 3a-d were used in the Diels-Alder reaction with N-methylmaleimide, and the adducts obtained 6a-d were oxidized to afford the xanthone-1,2,3-triazole dyads 7a-d. All the synthetic details as well as the structural characterization (by 1D and 2D NMR studies) of the new synthesised compounds will be presented and discussed.

Removal of antibiotic compounds by adsorption using glycerol-based carbon materials

This study is focused on the synthesis and application of glycerol-based carbon materials (GBCM200, GBCM300 and GBCM350) as adsorbents for the removal of the antibiotic compounds flumequine and tetracycline from aqueous solution. The synthesis enrolled the partial carbonization of a glycerol-sulfuric acid mixture, followed by thermal treatments under inert conditions and further thermal activation under oxidative atmosphere. The textural properties were investigated through N2 adsorption–desorption isotherms, and the presence of oxygenated groups was discussed based on zeta potential and Fourier transform infrared (FTIR) data. The kinetic data revealed that the equilibrium time for flumequine adsorption was achieved within 96 h, while for tetracycline, it was reached after 120 h. Several kinetic models, i.e., pseudo-first order, pseudo-second order, fractional power, Elovich and Weber–Morris models, were applied, finding that the pseudo-second order model was the most suitable for the fitting of the experimental kinetic data. The estimated surface diffusion coefficient values, Ds, of 3.88 and 5.06 10 14 m2 s 1, suggests that the pore diffusion is the rate limiting step of the adsorption process. Finally, as it is based on SSE values, Sips model well-fitted the experimental FLQ and TCN adsorption isotherm data, followed by Freundlich equation. The maximum adsorption capacities for flumequine and tetracycline was of 41.5 and 58.2 mg g 1 by GBCM350 activated carbon.

Analysis of foraminifera from the Tagus Estuary, Portugal

The distribution and microhabitat of living benthic fora- minifera (15 calcareous and 6 agglutinated) have been studied in two box cores from the Tagus Prodelta. Stable oxygen and carbon isotopes were analysed for eight different species from six surface samples from the Tagus Prodelta and Estuary. At the two box core stations, most of the living foraminifera were restricted to the oxygenated top cm of the sediment and generally show a shallow infaunal behavior. Those taxa are e.g. Rectuvigerina phlegeri, Stainforthia fusiformis and species of the genus Bolivina, which is the most abundant genus in the Tagus Prodelta. Infaunal species are found down to 10 cm depth, and some infaunal taxa, e.g. Bulimina marginata, Globobulimina auriculata and Nonionella turgida, inhabit the low oxic or anoxic sediments. The deep infaunal species are suggested to feed selectively, on refractory organic matter or on the bacterial stocks, while the opportunistic shallow infaunal species are believed to feed on fresh phytodetritus or labile organic matter. Our data show that there is a close connection between the concentration of foraminifera and the distribution of organic matter in the area. The highest abundance of living benthic foraminifera was found in sediments close to the Tagus river plume, where the sediments have relatively high organic carbon contents. The spatial distribution of the stable isotope values of different benthic foraminifera reflects the distribution of the low salinity and relatively high temperature water with high organic carbon fluxes within the Tagus Estuary.

Phosphorus components, trace metals and Ba excess of ODP Site 202-1237 sediments

We determined the sedimentary concentrations of phosphorus (P), barium (Ba), manganese (Mn), titanium (Ti), aluminum (Al), and uranium (U) for sediment samples from the southeast Pacific Nazca Ridge, Ocean Drilling Program Site 1237. This unique record extends to 31 Ma over 360 meters composite depth (mcd), recording depositional history as the site progressed eastward over its paleohistory. We sampled with a temporal resolution of ~0.2 m.y. throughout the sequence, equivalent to an average spacing of 1.63 m/sample. Concentrations of sequentially extracted components of P (oxide-associated, authigenic, organic, and detrital) increase toward the modern. Al/Ti ratios indicate that the background detrital source material is consistent with upper continental crust. U enrichment factors (U EFs) generally exceed crustal values and indicate slightly reducing environments. However, authigenic U precipitation can also be influenced by the organic carbon rain rate and may not be solely an indicator of redox conditions. Dramatic changes in Mn EFs at ~162 mcd, from values between 12 and 93 to values <12 after this depth, and a sharp color contact boundary lead us to believe that a paleoredox boundary from an oxygenated to a more reducing depositional environment occurred near this depth. Estimates of biogenic barite concentrations from a total sediment digestion technique (Ba excess) are greater than those from a barite extraction (Ba barite) for selected samples across the entire depth range. Applying a range of Ba/Ti ratios from different source materials to correct for detrital inputs does not change the lack of agreement with Ba barite concentrations. Reactive P (P reactive) concentrations (the sum of oxide-associated, authigenic, and organic P concentrations) increase toward the modern with values typically <12 µmol P/g from the base of our record through ~100 mcd, with a gradual increase to concentrations >15 µmol P/g. Ba excess follows the same general trends as Preactive, with concentrations <14 µmol Ba/g in the lower portion of the record to values >15 µmol Ba/g. Accumulation rate records of these proxies will be needed to infer paleoproductivity. P reactive/Ba excess ratios, an indicator of the relative burial of the nutrient P to organic carbon export, exhibit higher values, similar to modern, from the base of our record through ~180 mcd. The remainder of the record exhibits values lower than modern, indicating that organic carbon export to the sediments was higher relative to nutrient burial.

Sediment and organic matter properties, and molecular formula from sampling locations in the Northern Black Sea

Remineralization of organic matter in reactive marine sediments releases nutrients and dissolved organic matter (DOM) into the ocean. Here we focused on the molecular-level characterization of DOM by high-resolution Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) in sediment pore waters and bottom waters from contrasting redox regimes in the northern Black Sea with particular emphasis on nitrogen-bearing compounds to derive an improved understanding of the molecular transformations involved in nitrogen release. The number of nitrogen-bearing molecules is generally higher in pore waters than in bottom waters. This suggests intensified degradation of nitrogen-bearing precursor molecules such as proteins in anoxic sediments: No significant difference was observed between sediments deposited under oxic vs anoxic conditions (average O/C ratios of 0.55) suggesting that the different organic matter quality induced by contrasting redox conditions does not impact protein diagenesis in the subseafloor. Compounds in the pore waters were on average larger, less oxygenated, and had a higher number of unsaturations. Applying a mathematical model, we could show that the assemblages of nitrogen-bearing molecular formulas are potential products of proteinaceous material that was transformed by the following reactions: (a) hydrolysis and deamination, both reducing the molecular size and nitrogen content of the products and intermediates; (b) oxidation and hydration of the intermediates; and (c) methylation and dehydration.