Absolute configuration and conformation of the pure opioid antagonist (+)-2,9 alpha-dimethyl-5-(m-hydroxyphenyl)morphan.

(+)-2,9 alpha-Dimethyl-5-(m-hydroxyphenyl)morphan is the only phenylmorphan analog whose affinity for opioid kappa-receptors is greater than its affinity for opioid mu-receptors. Pharmacologically, the compound is a pure opioid antagonist devoid of agonist activity in in vivo assays of antinociception. The absolute configuration of the compound has been determined to be (1R,5S,9R) from an X-ray crystallographic study of the chloride salt. Thus, the absolute configuration corresponds to that of the atypical opioid agonist (-)-phenylmorphan while the weak atypical agonist (-)-2,9 alpha-dimethyl-5-(m- hydroxyphenyl)morphan corresponds to the potent morphine-like (+)-phenylmorphan. The preferred orientations of the phenyl ring for the two stereoisomers were determined using the molecular mechanics program MM2-87 and found to vary from that of the two parent compounds. The atypical properties of the two 9 alpha-methyl analogs is consistent with an opioid ligand model which proposes that morphine-like properties require a particular range of phenyl orientations. There was good agreement between the structure obtained from X-ray crystallography and computed with the MM2-87 program.     

Typologie des rivières oligotrophes du massif Ardennais (Belgique) par l'analyse multivariée de relevés de diatomées benthiques

Typology of oligotrophic streams of the Ardenne (Belgium) by multivariate analysis of benthic diatoms records.During three years, samples of water and algae has been taken in 72 stations distributed on the river system of the northern part of the Ardennes. 357 taxa of diatoms have been identified. Principal component analysis and cluster analysis, applied to diatoms counts, allowed to classify the records in a table comparable to phytosociological tables. A continuum in the data is an indication of the progressive modification of the communities according to the physico-chemical characteristics of the water. Ten diatoms assemblages have been described and this typology has been compared to some algal associations defined in the literature. In natural environments, these 10 assemblages are strictly correlated with the nature of the geological substrate and the physico-chemical characteristics of the water. The sensibility of the oligotrophic streams is revealed by the speedy changes in the communities after following a small modification of the environmental factors (seasonal variations or light eutrophication).

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Typologie des rivières oligotrophes du massif Ardennais (Belgique) par l'analyse multivariée de relevés de diatomées benthiques

     

Rapid reprogramming of haemoglobin structure‐function exposes multiple dual‐antimicrobial potencies

The intrinsic cytotoxicity of cell‐free haemoglobin (Hb) has hampered the development of reliable Hb‐based blood substitutes for over seven decades. Notably, recent evidence shows that the Hb deploys this cytotoxic attack against invading microbes, albeit, through an unknown mechanism. Here, we unraveled a rapid molecular reprogramming of the Hb structure‐function triggered by virulent haemolytic pathogens that feed on the haem‐iron. On direct contact with the microbe, the Hb unveils its latent antimicrobial potency, where multiple antimicrobial fragments are released, each harbouring coordinated ‘dual‐action centres’: microbe binding and pseudoperoxidase (POX) cycle activity. The activated Hb fragments anchor onto the microbe while the juxtaposed POX instantly unleashes a localized oxidative shock, killing the pathogen‐in‐proximity. This concurrent action conceivably restricts the diffusion of free radicals. Furthermore, the host astutely protects itself from self‐cytotoxicity by simultaneously releasing endogenous antioxidants. We found that this decryption mechanism of antimicrobial potency is conserved in the ancient invertebrate respiratory protein, indicating its fundamental significance. Our definition of dual‐antimicrobial centres in the Hb provides vital clues for designing a safer Hb‐based oxygen carrier blood substitute.     

CLIX: a search algorithm for finding novel ligands capable of binding proteins of known three-dimensional structure.

A computer algorithm, CLIX, capable of searching a crystallographic data-base of small molecules for candidates which have both steric and chemical likelihood of binding a protein of known three-dimensional structure is presented. The algorithm is a significant advance over previous strategies which consider solely steric or chemical requirements for binding. The algorithm is shown to be capable of predicting the correct binding geometry of sialic acid to a mutant influenza-virus hemagglutinin and of proposing a number of potential new ligands to this protein.     

Validation of the first step of the heuristic refinement method for the derivation of solution structures of proteins from NMR data

A new method for the analysis of NMR data in terms of the solution structure of proteins has been developed. The method consists of two steps: first a systematic search of the conformational space to define the region allowed by the initial set of experimental constraints, and second, the narrowing of this region by the introduction of additional constraints and optional refinement procedures. The search of the conformational space is guided by heuristics to make it computationally feasible. The method is therefore called the heuristic refinement method and is coded in an expert system called PROTEAN. The paper describes the validation of the first step of the method using an artificial NMR data set generated from the known crystal structure of sperm whale carbon monoxymyoglobin. It is shown that the initial search procedure yields a low-resolution structure of the myoglobin molecule, accurately reproducing its main topological features, and that the precision of the structure depends on the quality of the initial data set.     

Direct measurements of the dextran-dependent calcium uptake by rat peritoneal mast cells

The metallochromic indicator murexide has been used to monitor calcium concentration changes during the dextran-induced, phosphatidylserine-dependent degranulation of rat peritoneal mast cells. The dextran-induced Ca2+-uptake showed an absolute dependence on the presence of phosphatidylserine. The extent of Ca2+-uptake increased with phosphatidylserine in a concentration-dependent manner. At 25 degrees C the half-life of the uptake process equalled 35 +/- 5 s. Exposure of the mast cells to dextran in the presence of Ca2+, but in the absence of phosphatidylserine, desensitized the cells. The subsequent addition of phosphatidylserine failed to restore the Ca2+-uptake activity. However, the Ca2+-ionophore A23187 did promote Ca2+ uptake by the cells without PS.     

Explosive flower opening in ornithophily: a study of pollination mechanisms in some Central African Loranthaceae

In many ornithophilous Loranthaceae pollination is accompanied by an explosive opening of the flowers, and diverse mechanisms have evolved in different genera to bring this about. These are described for the African genera Erianthemum, Englerina, Tapinanthus, Globimetula, Vanwykia and Plicosepalus. In many genera tensions within the stamens cause the tubular corolla to split along the petal junctions to form window-like fenestrae. The flowers are pollinated mainly by sunbirds which insert their beaks through the fenestrae in search of the abundant nectar. This action causes the tube to split and the stamens to coil inwards explosively. In Globimetula and many species of Tapinanthus pigment is secreted along the edges of the specialized petal segments of the head, the spathulae. Probing along these secretory junctions causes the spathulae to reflex; further probing splits the corolla tube, and allows the stamens to coil inwards explosively. In Globimetula reflexure of the petals exposes the central column of stamens, between which secondary fenestrae are developed. In Plicosepalus curvature of the corolla tube is connected with a more specialized fenestral structure; flower opening is not explosive, and the open flowers continue to be visited regularly by sunbirds. In Vanwykia an early stage in the development of explosive flower-opening is found.     

Varicuothiols A and B,New Fungal Metabolites from Aspergillus versicolor with Anti‐Inflammatory Activities

Chemical examination of a coral‐associated fungus Aspergillus versicolor LZD‐44‐03 resulted in the isolation of two new compounds with the trivial names of varicuothiols A ( 1 ) and B ( 2 ) as a unique scaffold. Their structures were determined through extensive spectroscopic analyses in association with the modified Mosher's method and chemical conversion. Both 1 and 2 exhibited significant inhibition against LPS‐induced RAW24.7 cell proliferation, in association with the down regulation of nitrite production and cytokines (MCP‐1, IL‐6, and TNF‐α).     

Polymorphism of the SLB serologically defined (non-MHC) alloantigenic system in pigs

Six new antigenic specificities (designated SLB-2 to SLB-7) detectable with polyclonal reagents in the complement-dependent lymphocytotoxic test were determined in the SLB alloantigenic system of pigs. The SLB specificities (including the previously described SLB-1) are controlled by 13 alleles. An analysis of a further 121 double backcross matings (with 817 offspring) confirmed that the products of SLB alleles segregated independently of the SLA haplotypes (swine MHC). In addition, in 112 families of the same mating type (with 746 progeny) the map distance of the SLB locus and the highly polymorphic L blood group locus was found to be 7.82 centimorgans in the heterogametic sex and 12.57 centimorgans in the homogametic sex.