Effects of copper on algal communities at different current velocities

This study examines the influence of current velocity in the toxiceffect of copper in diatom-dominated biofilms grown in artificial channels.Effects on community structure, algal biomass and photosynthesis (carbonincorporation) caused by 15 g L^–1 of copperwere tested at contrasting (1 and 15 cm s^–1)velocities. Moreover, a possible threshold on the effect of copper on algalbiomass and photosynthesis related to current velocity was examined by usingprogressively increasing current velocity (1 to 50 cms^–1) at 15 g L^–1 Cu.Chlorophyll-a decreased ca. 50% as a result of addition of15 g L^–1 Cu. Chlorophyll decrease occurredearlier at 15 cm s^–1 than at 1 cms^–1 when adding 15 g L^–1Cu. Copper also caused a remarkable decrease in carbon incorporation(from 30 to ca. 50%), which was produced earlier at 15 cms^–1 (three days) than at 1 cms^–1 (seven days). Some taxa were affected by thecombination of copper and current velocity. Both Achnanthesminutissima and Stigeoclonium tenue becomedominant at 15 cm s^–1 in the presence of copper.Significant inhibition of algal growth in 15 g L^–1Cu occurred at low (1 cm s^–1) and highvelocities (50 cm s^–1), but not at intermediatevelocity (20 cm s^–1). The experiments indicatethat current velocity triggers the effect that copper has on diatom-dominatedbiofilms, and that the effect is more remarkable at low and high than atintermediate current velocities.     

Dmoesin controls actin-based cell shape and polarity during Drosophila melanogaster oogenesis

Ezrin, Radixin and Moesin (ERM) proteins are thought to constitute a bridge between the actin cytoskeleton and the plasma membrane (PM). Here we report a genetic analysis of Dmoesin, the sole member of the ERM family in Drosophila. We show that Dmoesin is required during oogenesis for anchoring microfilaments to the oocyte cortex. Alteration of the actin cytoskeleton resulting from Dmoesin mutations impairs the localization of maternal determinants, thus disrupting antero-posterior polarity. This study also demonstrates the requirement of Dmoesin for the specific organization of cortical microfilaments in nurse cells and, consequently, mutations in Dmoesin produce severe defects in cell shape.     

Primary hyperparathyroidism in an adult female olive baboon (Papio anubis)

During an annual physical examination, a middle-aged adult female olive baboon (Papio anubis) in the time-mated breeding colony at the Biologic Resources Laboratory at the University of Illinois at Chicago was found to have a high serum calcium value (> 12 mg/dl). To determine the cause of the hypercalcemia, additional diagnostic tests, including thoracic and abdominal radiographs and a parathyroid panel (parathyroid hormone (PTH), ionized calcium, and parathyroid hormone-related protein (PTH-rp) assays), were performed. The radiographs did not reveal lesions suggestive of neoplasia. A parathyroid panel was obtained twice. Both times the PTH (23.4 and 46.4 pmol/L, normal = 2.91 to 4.57 pmol/L) and ionized calcium (1.68 and 2.10 mmol/L, normal = 1.31 to 1.37 mmol/L) were increased above values for adult females with normal calcium concentration. A tentative diagnosis of primary hyperparathyroidism was made. After a gamma-radiation scan and magnetic resonance imaging of the neck were done, exploratory surgery was performed to identify and remove the affected gland. After gland removal, the baboon's serum calcium, PTH (1.6 pmol/L), and ionized calcium (1.59 mmol/L) values decreased. Results of histologic examination confirmed the diagnosis of benign solitary parathyroid adenoma.     

Selective alkylation and acylation of alpha and epsilon amino groups with PEG in a somatostatin analogue: tailored chemistry for optimized bioconjugates

The effects of the type and location of polymer grafting on the biological activity of different mono-PEG derivatives of the somatostatin analogue RC160 were evaluated. A chemical strategy to obtain mono-PEG alkylation or acylation of the peptide's alpha-terminal or lysil-epsilon primary amines was devised. Selective BOC protection of the two available primary amines, followed by reaction with two different PEG reagents and removal of the protecting group, was carried out. Chemical characterization, structural studies, and the evaluation of the biological activity of the bioconjugates synthesized allowed the identification of the one having characteristics more suitable for therapeutic application. This corresponds to the mono-epsilon-lysil-pegylated form, obtained by reductive alkylation, where the amine's positive charge is preserved. The results obtained suggest the importance of preliminary studies in the development of new polymer-peptide conjugates with improved pharmacological properties.     

Enzymatic acylation of di- and trisaccharides with fatty acids: choosing the appropriate enzyme,support and solvent

Enzymatic synthesis of fatty acid esters of di- and trisaccharides is limited by the fact that most biological catalysts are inactivated by the polar solvents (e.g. dimethylsulfoxide, dimethylformamide) where these carbohydrates are soluble. This article reviews the methodologies developed to overcome this limitation, namely those involving control over the reaction medium, the enzyme and the support. We have proposed the use of mixtures of miscible solvents (e.g. dimethylsulfoxide and 2-methyl-2-butanol) as a general strategy to acylate enzymatically hydrophilic substrates. We observed that decreasing the hydrophobicity of the medium (i.e. lowering the percentage of DMSO) the molar ratio sucrose diesters versus sucrose monoesters can be substantially enhanced. The different regioselectivity exhibited by several lipases and proteases makes feasible to synthesise different positional isomers, whose properties may vary considerably. In particular, the lipase from Thermomyces lanuginosus displays a notable selectivity for only one hydroxyl group in the acylation of sucrose, maltose, leucrose and maltotriose, compared with lipase from Candida antarctica. We have examined three immobilisation methods (adsorption on polypropylene, covalent coupling to Eupergit C, and silica-granulation) for sucrose acylation catalysed by T. lanuginosus lipase. The morphology of the support affected significantly the reaction rate and/or the selectivity of the process.     

Mast cells induce upregulation of P-selectin and intercellular adhesion molecule 1 on carotid endothelial cells in a new in vitro model of mast cell to endothelial cell communication

It is suggested that mast cells contribute to cell recruitment in inflammation through the upregulation of endothelial adhesion molecules. P-selectin and intercellular adhesion molecule(ICAM)-1 are two key adhesion molecules that have been associated indirectly with mast cell activity. The canine C2 mastocytoma cell line and primary cultures of canine carotid endothelial cells were used to establish a new in vitro model to help study the interaction between mast cells and endothelial cells. Carotid endothelial cells were incubated with mast cell mediators to uncover their effect on endothelial ICAM-1 and P-selectin expression. To assess the relative contributions of tumour necrosis factor (TNF)-alpha and histamine to such effect, an H1 antihistamine and a TNF-alpha blocking antibody were used. Prior to activation by mast cell mediators, P-selectin was expressed only within the cytoplasm, and ICAM-1 was constitutively expressed on the surface of the canine carotid endothelial cells. Both adhesion molecules were enhanced significantly and strongly upon mast cell activation at various time points. Unstored TNF-alpha was fully responsible for ICAM-1 upregulation. P-selectin was up-regulated by both preformed and newly synthesized mast cell mediators, but neither histamine nor TNF-alpha accounted for such an effect. Therefore,a new model is proposed in which the pro-inflammatory effect of mast cells on endothelial cells can be studied in vitro.In this model, it has been demonstrated that only TNF-alpha accounts for the overexpression of ICAM-1 induced by mast cells, and that mast cells up-regulate P-selectin expression through a histamine-independent mechanism.     

A chemical,genetic, and structural analysis of the nuclear bile acid receptor FXR

The farnesoid X receptor (FXR) functions as a bile acid (BA) sensor coordinating cholesterol metabolism, lipid homeostasis, and absorption of dietary fats and vitamins. However, BAs are poor reagents for characterizing FXR functions due to multiple receptor independent properties. Accordingly, using combinatorial chemistry we evolved a small molecule agonist termed fexaramine with 100-fold increased affinity relative to natural compounds. Gene-profiling experiments conducted in hepatocytes with FXR-specific fexaramine versus the primary BA chenodeoxycholic acid (CDCA) produced remarkably distinct genomic targets. Highly diffracting cocrystals (1.78 A) of fexaramine bound to the ligand binding domain of FXR revealed the agonist sequestered in a 726 A(3) hydrophobic cavity and suggest a mechanistic basis for the initial step in the BA signaling pathway. The discovery of fexaramine will allow us to unravel the FXR genetic network from the BA network and selectively manipulate components of the cholesterol pathway that may be useful in treating cholesterol-related human diseases.     

Development of specific fluorescent oligonucleotide probes for in situ identification of wine lactic acid bacteria

A rapid method for the identification of lactic acid bacteria (LAB) from wine has been developed. This method is based on fluorescence in situ hybridisation (FISH), using fluorescent oligonucleotide probes, homologous to 16S rDNA of those species of LAB commonly found in wines. The protocol for the specific detection of these bacteria was established through the hybridisation of 36 reference strains. The specificity of the probes was evaluated by using pure cultures. Probes were used to identify species in different wines, making it evident that direct identification and quantification from natural samples without culturing is also possible. The results show that FISH is a promising technique for the rapid identification of LAB, allowing positive identification in a few hours (4-16 h).     

Shifts in immunoglobulin (IgG,IgM and IgA) levels in the milk of southern elephant seals,at Potter Peninsula,King George Island,Antarctica

We quantified immunoglobulins (Ig) in mammary secretions of 12 female southern elephant seals ( Mirounga leonina) at King George Island, Antarctica, using single radial immunodiffusion on agarose plates. Seals were chemically immobilized for milk sample collection at four time points during the 3- to 4-week suckling period. All three major mammalian immunoglobulins (IgG, IgM, IgA) were detected in southern-elephant seal milk. Total immunoglobulin levels and the ratio of Ig subclasses varied throughout the suckling period. Total immunoglobulin levels were highest on the 1st day of lactation, when they represented over 57% of the mean total protein concentration of the milk, and declined steadily throughout lactation, representing approximately 40% of the mean total protein concentration at the end of the suckling period. IgG was the most abundant immunoglobulin class (85.93-93.78% of total milk immunoglobulins), followed by IgM (6.06-13.93%), and IgA (0.14-0.23%). There was no significant difference in IgA levels throughout the suckling period. IgG levels were significantly lower during the second stage (3-6 days post-parturition) than during the first, third or fourth stages (1, 13-15, and 19-25 days post-parturition, respectively). IgM levels were highest during the first stage of lactation; these values were significantly higher than levels measured during the second, third and fourth stages of lactation. Transfer of passive immunity from female to offspring in other mammalian species is correlated with the subclass of immunoglobulin secreted in the milk; species acquiring passive immunity in utero, via the placenta, secrete a preponderance of IgA, whereas species acquiring immunity post-partum, via lacteal secretions and gut resorption, secrete a preponderance of IgG. The Ig patterns and concentrations observed in our study of southern elephant seals are consistent with an important role of post-partum transmission of passive immunity during the pinniped lactation period.     

Novel maltotriose esters enhance biodegradation of Aroclor 1242 by Burkholderia cepacia LB400

The objective of this research was to evaluate the effect of enzymatically synthesized maltotriose fatty acid monoesters (Ferrer, M., et al. 2000 Tetrahedron 56, 4053–4061) on Aroclor 1242 solubilization and biodegradation. Three forms of the surfactant, laurate, palmitate and stearate monoester, were tested. Potential enhancement of solubilization of hydrophobic substances mediated by these non-ionic surfactants was exploited in this study. A polychlorinated biphenyl (PCB) degrading organism, Burkholderia cepacia LB400, was also selected. It was found that all surfactants were effective in solubilizing Aroclor 1242 but the rate of Aroclor 1242 biodegradation proceeded rapidly only in the presence of 6-O-palmitoylmaltotriose. For example, the addition of 48 mg 6-O-palmitoylmaltotriose/l increased the apparent solubility from 140 to 305 g/l. As a result, only 8% of the Aroclor remained at the end of 24 h incubation. In contrast, 49.2% of the Aroclor 1242 remained in the absence of surfactant. It appears that maltotriose fatty acid monoesters can significantly increase the bioavailability, and thereby accelerate the biodegradation of highly chlorinated PCBs, particularly Aroclor 1242, by Burkholderia cepacia LB400. The possibility of obtaining these biodegradable surfactants with high yield, easy recovery and high purity by using a new enzymatic methodology, makes maltotriose esters available for bioremediation purposes.