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Lectins from plants of the Diocleinae subtribe often exhibit specificity towards mannose/glucose and derived sugars, with some plants also displaying a second lectin specific to lactose/GalNAc. Here, we present a novel lectin from Collaea speciosa, named CsL, that displays specificity for GlcNAc/glucose. The lectin was extracted from Collaea speciosa seeds and purified by a single chromatographic step on a Sephadex G-50 matrix. In solution, the lectin appears as a dimeric protein composed of 25 kDa monomers. The protein is stable at pH 7-8 and dependent on divalent cations. CsL maintained its agglutination activity after heating to 90 °C for 1 h. Glycan array studies revealed that CsL binds to N-glycans with terminal GlcNAc residues, chitobiose and chitotriose moieties. The partial amino acid sequence of the lectin is similar to that of some lactose-specific lectins from the same subtribe. In contrast to other ConA-like lectins, CsL is not toxic to Artemia. Because of its remarkably different properties and specificity, this lectin could be the first member of a new group inside the Diocleinae lectins.

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Iron metallodrugs comprise mineral supplements, anti-hypertensive agents and, more recently, magnetic nanomaterials, with both therapeutic and diagnostic roles. As biologically-active metal compounds, concern has been raised regarding the impact of these compounds when emitted to the environment and associated ecotoxicological effects for the fauna. In this work we assessed the relative stability of several iron compounds (supplements based on glucoheptonate, dextran or glycinate, as well as 3,5,5-trimethylhexanoyl (TMH) derivatives of ferrocene) against high affinity models of biological binding, calcein and aprotransferrin, via a fluorimetric method. Also, the redox-activity of each compound was determined in a physiologically relevant medium. Toxicity toward Artemia salina at different developmental stages was measured, as well as the amount of lipid peroxidation. Our results show that polymer-coated iron metallodrugs are stable, non-redox-active and non-toxic at the concentrations studied (up to 300 µM). However, TMH derivatives of ferrocene were less stable and more redox-active than the parent compound, and TMH-ferrocene displayed toxicity and lipid peroxidation to A. salina, unlike the other compounds. Our results indicate that iron metallodrugs based on polymer coating do not present direct toxicity at low levels of emission; however other iron species (eg. metallocenes), may be deleterious for aquatic organisms. We suggest that ecotoxicity depends more on metal speciation than on the total amount of metal present in the metallodrugs. Future studies with discarded metallodrugs should consider the chemical speciation of the metal present in the composition of the drug.

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Lectins are sugar-binding proteins widely distributed in nature with many biological functions. Although many lectins have a remarkable biotechnological potential, some of them can be cytotoxic. Thus, the aim of this study was to assess the toxicity of five lectins, purified from seeds of different species of Canavalia genus. In order to determine the toxicity, assays with Artemia nauplii were performed. In addition, a fluorescence assay was carried out to evaluate the binding of lectins to Artemia nauplii. In order to verify the relationship between the structure of lectins and their cytotoxic effect, structural analysis was carried out to evaluate the volume of the carbohydrate recognition domain (CRD) of each lectin. The results showed that all lectins exhibited different toxicities and bound to a similar area in the digestive tract of Artemia nauplii. Concerning the structural analysis, differences in spatial arrangement and volume of CRD may explain the variation of the toxicity exhibited by each lectin. To this date, this is the first study that establishes a link between toxicity and structure of CRD from Diocleinae lectins.

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Using differential scanning calorimetry we demonstrated the presence of biological glasses and measured the glass transition temperatures (Tg) in dry encysted gastrula embryos (cysts) of the brine shrimp, Artemia, from eleven different locations, two of which provided cysts from parthenogenetic animals. Values for Tg were highest, by far, in Artemia franciscana cysts from the Mekong Delta, Vietnam (VN), these cysts having been produced from previous sequential inoculations into growth ponds of cysts from the San Francisco Bay, California, USA. Tg values for three groups of A. franciscana cysts were significantly higher than those of other cysts (except those of Artemia persimilis) studied here, as well as all other desiccation-tolerant animal systems studied to date. We also measured three stress proteins (hsc70, artemin and p26) in all these cysts as well as the total alcohol soluble carbohydrates (ASC), about 90% of which is the disaccharide trehalose, a known component of biological glasses. We interpret the results in terms of mechanisms involved with desiccation tolerance and, to some extent, with thermal conditions at the sites of cyst collection.

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The objective of this investigation was to assess the transfer of lead (Pb) along an experimental, four-level food chain: Tetraselmis suecica (phytoplankton) â†’ Artemia franciscana (crustacean, brine shrimp) â†’ Litopenaeus vannamei (crustacean, white shrimp) â†’ Haemulon scudderi (fish, grunt). T. suecica was exposed to a sublethal dose of Pb in solution and then used as the base of a marine food chain. Significant differences in Pb concentrations were found between exposed organisms of the different trophic levels and the control. Particularly, Pb concentrations in fish of the simulated trophic chain were two-to three times higher in the exposed specimens than in the control. Levels of Pb in phytoplankton showed a substantial increase with respect to the solution (level I), with bioconcentration factors averaging from 930 to 3630. In contrast, a strong decrease in Pb concentration from phytoplankton to zooplankton (level II) and from zooplankton to shrimp tissues (level III) was evidenced by bioaccumulation factors <1. Despite the decrease in the assimilation efficiency of metal transfer observed in these two predators, Pb concentration in the grunt fish (level IV) was higher than in the shrimp (level III) (bioaccumulation factor >1.0). Some of the added Pb is transferred from the phytoplankton along the food chain, thus producing a net accumulation of Pb mainly in fish and, to a lesser extent, in shrimp tissues. Because Pb is one of the most pervasive contaminants in coastal ecosystems, its transference by way of diet and potential net accumulation in higher predators is of ecologic importance for marine life. In addition, because shrimp and adult Haemulon scudderi are commercially important resources, this issue is of particular relevance to the safety of marine products.

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Schistosomiasis or bilharzia, which affects millions of people living in Africa, Asia and Latin America, is closely associated with certain species of aquatic snails. One way of attacking the disease is to eradicate the host snails. Molluscicidal activities of natural compounds are especially important in the widespread control of this tropical disease. As part of our search for natural compounds with molluscicidal properties for the vector control of schistosomiasis, we are now evaluating for the first time the toxicity of the plant lectins from Canavalia brasiliensis (ConBr), Cratylia floribunda (CFL), Dioclea guianensis (Dgui), Dioclea grandiflora (DGL) and Dioclea virgata (Dvir) to Biomphalaria glabrata Say and Artemia salina Leach. Results indicate that all the samples were toxic to A. salina Leach, some of them with values of lethal concentration that kills 90% of the population (LC(90))<10 microg mL(-1). They are also active against B. glabrata Say, killing 100% of adult snails, at a concentration of 50 microg mL(-1). The lectins CFL and Dgui possess properties lethal to mollusks, with values of LC(90)=50.3 microg mL(-1) and LC(90)=41.0 microg mL(-1), respectively.

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The effect of metal ions, ferric ion (Fe3+) and molybdenum ion (Mo6+) on the denitrification process of Paracoccus pantotrophus P16 grown under saline conditions was investigated. Results revealed that the dosages of added Fe3+ and Mo6+ significantly accelerated nitrate utilization and nitrite accumulation. Enzymatic studies revealed that the membrane-bound nitrate reductase and the periplasmic nitrite reductase had activities of 998 +/- 28 and 373 +/- 18 nmol (mg protein)-1 min-1, respectively after growing Paracoccus pantotrophus P16 in medium supplemented with 1.5 micron M Fe3+. If provided with 1.5 micron M Fe3+and 2.4 micron M Mo6+, the membrane-bound nitrate reductase activity increased to 6,223 +/- 502 nmol (mg protein)-1 min-1 and the periplasmic nitrite reductase was 344 +/- 20 nmol (mg protein)-1 min-1. The results indicated that an addition of Fe3+ and Mo6+ led to an overstimulation of nitrate reductase activity as compared with nitrite reductase activity. When glucose was supplied, the minimal ratio of carbon per nitrate (C/N) was 2.31 mg C/mg NO3--N with denitrification yield of 0.45 g NO3--N/g C. Addition of ethanol instead of glucose, the minimal ratio of C/N was 1.15 mg C/mg NO3--N with denitrification yield of 1.08 g NO3--N/g C.

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Nonylphenol is a metabolic intermediate from the microbial transformation of detergents used worldwide. While nonylphenol shows some acute toxicity, it is also able to mimic important hormones resulting in the disruption of several processes by interfering with the signals that control the overall physiology of the organism. The effect of the pollutant nonylphenol (NP) through the trophic chain was studied. Microalgae Isochrysis galbana was able to bioconcentrate NP 6940 times, where 77% of initial NP (100microgl(-1)) is accumulated intracellularly after 1-h incubation. Crustacean Artemia fransiscana showed 25% higher growth when fed with NP-rich algae. However, Artemia metabolized almost all NP ingested and only traces of NP could be found in the organism, eliminating future NP effects. Zebrafish (Brachydanio rerio) were affected by the presence of 171microgg(-1) of NP in the diet, showing higher levels of the hormone vitellogenin and lower levels of cytochrome P450 activity. These results showed that organisms placed in the first level of trophic chain are able to significantly bioconcentrate the pollutant and endocrine disruptor NP. These grassed organisms affect the growth of crustacean. Moreover, the organisms placed on the top of some trophic chains, such as fish, could be affected by the presence of NP in their food, in both the hormone levels and metabolic enzymes. This work shows that the environmental presence of NP should be considered as a risk for the organisms living in an ecosystem.

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The agglutinating activity of the hemolymph of Litopenaeus schmitti is insensitive to calcium and specific for acetylated sugars, particularly sialic acid (Neu5Ac) and O-sialoglycoconjugates (bovine submaxillary mucin) and has varying specificity for different LPS, which may suggest a putative role in microorganism recognition. Affinity chromatography on fetuin-agarose of the agglutinin resulted in a 220 kDa band (lectin), and a 82.5 kDa band, which probably is hemocyanin. The 220 kDa protein consists of 31 and 34 kDa subunits, suggesting that this lectin is multimeric. The lectin molecular mass was estimated by gel filtration to be 153+/-10 kDa. The hemolymph of L. schmitti comprises at least another soluble lectin, with distinct chemical and carbohydrate specificity than the 220 kDa lectin.

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