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Ecological limitations on aquatic mosquito predator colonization in the urban environment.

Urban malaria cases are becoming common in Africa as more people move into cities and industrialization proceeds. While many species of Anopheles mosquitoes vector malaria in rural areas, only a few are found within cities. The success of anthropophilic species in cities, such as members of the An. gambiae complex, may be explained by limitations on colonization by predator species in urban environments. Habitats that are temporal or structurally simple have lower predator survivorship in a variety of ecosystems, but these have not been investigated previously in an urban area. Areas within and around the Kenyan coastal town of Malindi were previously sampled for the presence of standing water using a geographic sampling strategy with probability proportional to size sampling of planned well-drained, unplanned poorly-drained, planned poorly-drained, unplanned well-drained, and peri-urban locations. Standing aquatic habitats in these areas were reassessed. During monthly sampling, presence/absence of mosquitoes and predator taxa were noted, as were ecological habitat variables: structural complexity and presence of water. Lambda statistics were calculated to associate predator guilds, habitat types, location variables, and ecological variables. All predator guilds found in habitats were strongly associated with habitat type, as were the structural complexity and temporal nature of the habitats. Types of habitat were heterogeneously distributed throughout Malindi, with swimming pools as a common habitat type in planned urban areas and tire track pools a common habitat type in peri-urban areas of Malindi. Predator colonization of aquatic habitats in Malindi was strongly influenced by habitat type, and not associated with location characteristics. Ecological variables were affected by the type of habitats, which are co-associated with planning and drainage in Malindi. While habitat types are distributed heterogeneously within Malindi, habitats with low predation pressure are available for mosquito colonization in both urban and peri-urban areas. The temporal, peri-urban tire track pools and the structural simplicity of urban swimming pools may discourage predator colonization, thereby increasing the probability of malaria vectors in these areas of Malindi. Future studies should evaluate habitats for use in malaria surveillance and experimentally test the effects of structural complexity and temporal nature of urban habitats on the densities of mosquito larvae and their aquatic predators.

Animals↗

Influence of polymer architecture on the structure of complexes formed by PEG-tertiary amine methacrylate copolymers and phosphorothioate oligonucleotide.

The influence of polymer structure on the characteristics of complexes of a phosphorothioate antisense oligonucleotide (ISIS 5132) was studied, using well-defined cationic copolymers based on 2-(dimethylamino) ethyl methacrylate (DMAEMA) and poly(ethylene glycol) (PEG). The three related copolymer structures were: DMAEMA-PEG (a diblock copolymer) DMAEMA-OEGMA 7 (a brush-type copolymer), DMAEMA-stat-PEGMA (a comb-type copolymer); each of these were examined together with DMAEMA homopolymer, which served as a control. The results revealed that all the polymers exhibited good binding ability with the oligonucleotide (ON). Interestingly, the comb-type polymer DMAEMA-stat-PEGMA demonstrated the highest binding ability and DMAEMA homopolymer the lowest, as judged by a dye displacement assay. DMAEMA homopolymer produced large agglomerates of smaller individual complexes as observed by optical density, photon correlation spectroscopy and transmission electron microscopy studies. In contrast, two PEG-block copolymers, DMAEMA-PEG and DMAEMA-OEGMA 7, formed compact complexes of 80-150 nm which had good long-term colloidal stability. This is attributed to the steric stabilisation effect of the PEG chains on the ON-copolymer complexes. These two copolymers are believed to form complexes with ON that have a micellar structure. Comb-type DMAEMA-stat-PEGMA copolymer formed highly soluble complexes with the ON that did not phase separate from the buffer solution. This study clearly demonstrates that varying the copolymer architecture allows access to a range of ON complexes. In vitro cytotoxicity experiments on HepG2 cells showed that all of the tertiary amine methacrylate copolymers displayed lower cytotoxicity than the control poly(L-lysine).

Amines↗

Structure of complex of synthetic HIV-1 protease with a substrate-based inhibitor at 2.3 A resolution.

The structure of a complex between a peptide inhibitor with the sequence N-acetyl-Thr-Ile-Nle-psi[CH2-NH]-Nle-Gln-Arg.amide (Nle, norleucine) with chemically synthesized HIV-1 (human immunodeficiency virus 1) protease was determined at 2.3 A resolution (R factor of 0.176). Despite the symmetric nature of the unliganded enzyme, the asymmetric inhibitor lies in a single orientation and makes extensive interactions at the interface between the two subunits of the homodimeric protein. Compared with the unliganded enzyme, the protein molecule underwent substantial changes, particularly in an extended region corresponding to the "flaps" (residues 35 to 57 in each chain), where backbone movements as large as 7 A are observed.

Amino Acid Sequence↗

Biomimetic copper(I)--CO complexes: a structural and dynamic study of a cali.

Four novel calix[6]arene-based cuprous complexes are described. They present a biomimetic tris(imidazole) coordination core associated with a hydrophobic cavity that wraps the apical binding site. Each differs from the other by the methyl or ethyl substituents present on the phenoxyl groups (OR1) and on the imidazole arms (NR2) of the calix[6]arene structure. In solution, stable CO complexes were obtained. We have investigated their geometrical and dynamic properties with respect to the steric demand. IR and NMR studies revealed that, in solution, these complexes adopted two distinct conformations. The preferred conformation was dictated only by the size of the OR1 group. When R1 was an ethyl group, the complex preferentially adopted a flattened C3-symmetrical structure. The corresponding helical enantiomers were in conformational equilibrium, which, however, was slow on the 1H NMR time scale at -80 degrees C. When R1 was a methyl group, the low-temperature NMR spectra revealed the partial inclusion of one tBu group. The complex wobbled between three dissymmetric but equivalent conformations. Hence, small differences in the steric demand of the calixarene's skeleton changed the geometry and dynamics of the system. Indeed, this supramolecular control was promoted by the strong conformational coupling between the metal center and the host structure. Interestingly, this was not only the result of a covalent preorganization, but also stemmed from weak interactions within the hydrophobic pocket. The vibrational spectra of the bound CO were revealed to be a sensitive gauge of this supramolecular behavior, similar to copper proteins in which allosteric effects are common.

Carbon Monoxide↗

New tetranuclear Cu(II) complexes: synthesis, structure, and magnetic properties.

The synthesis, structure, and magnetic properties of two new tetranuclear Cu(II) complexes containing N,N,N',N'-tetraethylpyridine-2,6-dithiocarboxamide (S-dept) of formula [Cu(2)Cl(2)(mu-S-dept)(2)][Cu(2)Cl(4)(mu-Cl)(2)] (1) and [Cu(2)(mu-Cl)(2)(S-dept)(2)][CuCl(3)(EtOH)](2) (2) are reported. Their X-ray crystal structures reveal that the complexes are composed of anionic and cationic dimers, that in both cases contain the metal centers which interact via Coulombic and/or hydrogen bonding interactions. In both cases, the Cu centers in the anionic moieties adopt a slightly distorted tetrahedral geometry whereas for the cationic moieties they adopt a square-pyramidal type of geometry. Magnetic susceptibility data show that compounds 1 and 2 present an overall antiferromagnetic behavior arising from the contribution of both anionic and cationic moieties. For 1, the best fit obtained gave J(1) = -2.62 +/- 0.19 cm(-1), J(2) = -19.54 +/- 0.47 cm(-1), and g(2) = 2.164 +/- 0.004 cm(-1) (R = 8.28 x 10(-5)) whereas for 2 it gave J(1) = 4.48 +/- 2.73 cm(-1), g(1) = 2.20 +/- 0.03, J(2) = -11.26 +/- 2.01 cm(-1), and g(2) = 2.10 +/- 0.03 (R = 1.15 x 10(-4)). The nature of the superexchange pathways in 1 and 2 is discussed on the basis of structural, magnetic, and molecular orbital considerations. Theoretical calculations are performed at the extended Huckel level in order to obtain their molecular orbitals and energies using their crystallographic data.

Journal Article↗

A complex secondary structure in U1A pre-mRNA that binds two molecules of U1A protein is required for regulation of polyadenylation.

The human U1A protein-U1A pre-mRNA complex and the relationship between its structure and function in inhibition of polyadenylation in vitro were investigated. Two molecules of U1A protein were shown to bind to a conserved region in the 3' untranslated region of U1A pre-mRNA. The secondary structure of this region was determined by a combination of theoretical prediction, phylogenetic sequence alignment, enzymatic structure probing and molecular genetics. The U1A binding sites form (part of) a complex secondary structure which is significantly different from the binding site of U1A protein on U1 snRNA. Studies with mutant pre-mRNAs showed that the integrity of much of this structure is required for both high affinity binding to U1A protein and specific inhibition of polyadenylation in vitro. In particular, binding of a single molecule of U1A protein to U1A pre-mRNA is not sufficient to produce efficient inhibition of polyadenylation.

Base Sequence↗

Structural characterization of photosystem II complex from red alga Porphyridium cruentum retaining extrinsic subunits of the oxygen-evolving complex.

The structure of photosystem II (PSII) complex isolated from thylakoid membranes of the red alga Porphyridium cruentum was investigated using electron microscopy followed by single particle image analysis. The dimeric complexes observed contain all major PSII subunits (CP47, CP43, D1 and D2 proteins) as well as the extrinsic proteins (33 kDa, 12 kDa and the cytochrome c(550)) of the oxygen-evolving complex (OEC) of PSII, encoded by the psbO, psbU and psbV genes, respectively. The single particle analysis of the top-view projections revealed the PSII complex to have maximal dimensions of 22 x 15 nm. The analysis of the side-view projections shows a maximal thickness of the PSII complex of about 9 nm including the densities on the lumenal surface that has been attributed to the proteins of the OEC complex. These results clearly demonstrate that the red algal PSII complex is structurally very similar to that of cyanobacteria and to the PSII core complex of higher plants. In addition, the arrangement of the OEC proteins on the lumenal surface of the PSII complex is consistent to that obtained by X-ray crystallography of cyanobacterial PSII.

Algal Proteins↗

Recognition of IgG by Fcgamma receptor. The role of Fc glycosylation and the binding of peptide inhibitors.

Recently determined crystal structures of the complex between immunoglobulin constant regions (Fc) and their Fc-respective receptors (FcR) have revealed the detailed molecular interactions of this receptor-ligand pair. Of particular interest is the contribution of a glycosylation at Asn(297) of the C(H)2 domain of IgG to receptor recognition. The carbohydrate moieties are found outside the receptor.Fc interface in all receptor.Fc complex structures. To understand the role of glycosylation in FcR recognition, the receptor affinities of a deglycosylated IgG1 and its Fc fragment were determined by solution binding studies using surface plasmon resonance. The removal of carbohydrates resulted in a non-detectable receptor binding to the Fc alone and a 15- to 20-fold reduction of the receptor binding to IgG1, suggesting that the carbohydrates are important in the function of the FcgammaRIII. Structurally, the carbohydrates attached to Asn(297) fill the cavity between the C(H)2 domains of Fc functioning equivalently as a hydrophobic core. This may stabilize a favorable lower hinge conformation for the receptor binding. The structure of the complex also revealed the dominance of the lower hinge region in receptor.Fc recognition. To evaluate the potential of designing small molecular ligands to inhibit the receptor function, four lower hinge peptides were investigated for their ability to bind to the receptor FcgammaRIII. These peptides bind specifically to FcgammaRIII with affinities 20- to 100-fold lower than IgG1 and are able to compete with Fc in receptor binding. The results of peptide binding illustrate new ways of designing therapeutic compounds to block Fc receptor activation.

Amino Acid Sequence↗

32-Indolyl ether derivatives of ascomycin: three-dimensional structures of complexes with FK506-binding protein.

32-Indole ether derivatives of tacrolimus and ascomycin retain the potent immunosuppressive activity of their parent compounds but display reduced toxicity. In addition, their complexes with the 12-kDa FK506-binding protein (FKBP) form more stable complexes with the protein phosphatase calcineurin, the molecular target of these drugs. We have solved the three-dimensional structures of the FKBP complexes with two 32-indolyl derivatives of ascomycin. The structures of the protein and the macrolide are remarkably similar to those seen in the complexes with tacrolimus and ascomycin. The indole groups project away from the body of the complex, and multiple conformations are observed for the linkage to these groups as well as for a nearby peptide suggesting apparent flexibility in these parts of the structure. Comparison of these structures with that of the ternary complex of calcineurin, FKBP, and tacrolimus suggests that the indole groups interact with a binding site comprising elements of both the calcineurin alpha- and beta-chains and that this interaction is responsible for the increased stability of these complexes.

Crystallography, X-Ray↗

SU9516: biochemical analysis of cdk inhibition and crystal structure in complex with cdk2.

SU9516 is a 3-substituted indolinone compound with demonstrated potent and selective inhibition toward cyclin dependent kinases (cdks). Here, we describe the kinetic characterization of this inhibition with respect to cdk2, 1, and 4, along with the crystal structure in complex with cdk2. The molecule is competitive with respect to ATP for cdk2/cyclin A, with a K(i) value of 0.031 microM. Similarly, SU9516 inhibits cdk2/cyclin E and cdk1/cyclin B1 in an ATP-competitive manner, although at a 2- to 8-fold reduced potency. In contrast, the compound exhibited non-competitive inhibition with respect to ATP toward cdk4/cyclin D1, with a 45-fold reduced potency. The X-ray crystal structure of SU9516 bound to cdk2 revealed interactions between the molecule and Leu83 and Glu81 of the kinase. This study should aid in the development of more potent and selective cdk inhibitors for potential therapeutic agents.

Adenosine Triphosphate↗

Porcine colonic lymphoglandular complex: distribution, structure, and epithelium.

Lymphoepithelium and cells specialized for uptake and transport of foreign matter are characteristic of antigen sampling organs, including lymphoglandular complexes (LGCs). Distribution, histologic structure, and epithelial ultrastructure of colonic lymphoglandular complexes were determined in 5- to 13-week-old pigs. LGCs averaged 1,231 in number per colon, displayed a characteristic distribution pattern, and were most evenly distributed in colons of older pigs. LGCs consisted of well-defined submucosal masses composed of lymphatic nodules and internodular lymphoid tissue penetrated by radially branching extensions of mucosal glands. Epithelial diverticula of each LGC entered the submucosa as a group through a circular collar derived from the muscularis mucosae. LGC epithelium contained goblet cells, cuboidal and columnar enterocytes, enteroendocrine cells, individual and clustered intraepithelial leukocytes, and cells morphologically compatible with follicle-associated epithelial cells/M cells. We regard the colonic LGC as a distinct mucosal lymphoid organ and suggest a significant role for it in local and systemic immune responses. The porcine colonic LGC may serve as a model for the human LGC.

Animals↗

Cyano-bridged Mn(III)3M(III) (M(III) = Fe, Cr) complexes: synthesis, structure, and magnetic properties.

Two cyano-bridged tetranuclear complexes composed of Mn(III) salen (salen = N,N'-ethylene bis(salicylideneiminate)) and hexacyanometalate(III) (M = Fe, Cr) in a stoichiometry of 3:1 have been selectively synthesized using {NH2(n-C12H25)2}3[M(III)(CN)6] (M(III) = Fe, Cr) starting materials: [{Mn(salen)(EtOH)}3{M(CN)6}] (M = Fe, 1; Cr, 2). Compounds 1 and 2 are isostructural with a T-shaped structure, in which [M(CN)6]3- assumes a meridional-tridentate building block to bind three [Mn(salen)(EtOH)]+ units. The strong frequency dependence and observation of hysteresis on the field dependence of the magnetization indicate that 1 is a single-molecule magnet.

Journal Article↗

Physiological mechanisms of flooding (implosion) therapy.

Desensitization of psychological and physiological complex structures may be the most important element of flooding treatment. The implosive sessions are assumed to represent a supramaximal stimulation of pathologically excited and inert complex structures resulting in protective inhibition, irradiation of excitation, reduction of the excitation and inertness, and a decrease of the overshooting autonomic reactivity of the complex structures, leading to reduction of anxiety, aggression, and other pathologically increased feelings. Advantages such as stronger and improved flooding can be achieved by a flooding in hypnosis. The therapeutic indications go beyond the usual treatment of phobic states. In order to establish the psychological and physiological mechanisms in implosion there is a need for psychophysiological investigations. However, much is unknown about mechanisms. Controlled comparisons with other treatments give limited answers. Perhaps an international case history bank might establish which clinical conditions might benefit by technical modifications of flooding.

Animals↗

Technetium carboxylate complexes--II. Structural and chemical studies.

Formate, acetate, and pivalate based complexes of technetium have been prepared by the electrochemical reduction of pertechnetate in the appropriate carboxylate buffer. Also, chemical synthetic routes to these complexes have been explored using technetium chloro compounds of known oxidation states as starting materials. The technetium carboxylate complexes were selectively examined by subtractively normalized interfacial Fourier transform infrared, electron spin resonance, and electronic spectroscopy, electrochemistry, electrophoresis, and anion exchange chromatography. These complexes have been proposed to be metal-metal bonded dioxo bridged ditechnetium(IV) species with axial and bridged carboxylate ligands.

Carboxylic Acids↗

Crystal structures of complexes between aminoglycosides and decoding A site oligonucleotides: role of the number of rings and positive charges in the specific binding leading to miscoding.

The crystal structures of six complexes between aminoglycoside antibiotics (neamine, gentamicin C1A, kanamycin A, ribostamycin, lividomycin A and neomycin B) and oligonucleotides containing the decoding A site of bacterial ribosomes are reported at resolutions between 2.2 and 3.0 A. Although the number of contacts between the RNA and the aminoglycosides varies between 20 and 31, up to eight direct hydrogen bonds between rings I and II of the neamine moiety are conserved in the observed complexes. The puckered sugar ring I is inserted into the A site helix by stacking against G1491 and forms a pseudo base pair with two H-bonds to the Watson-Crick sites of the universally conserved A1408. This central interaction helps to maintain A1492 and A1493 in a bulged-out conformation. All these structures of the minimal A site RNA complexed to various aminoglycosides display crystal packings with intermolecular contacts between the bulging A1492 and A1493 and the shallow/minor groove of Watson-Crick pairs in a neighbouring helix. In one crystal, one empty A site is observed. In two crystals, two aminoglycosides are bound to the same A site with one bound specifically and the other bound in various ways in the deep/major groove at the edge of the A sites.

Adenine↗

Involvement of bacterial migration in the development of complex multicellular structures in Pseudomonas aeruginosa biofilms.

Detailed knowledge of the developmental process from single cells scattered on a surface to complex multicellular biofilm structures is essential in order to create strategies to control biofilm development. In order to study bacterial migration patterns during Pseudomonas aeruginosa biofilm development, we have performed an investigation with time-lapse confocal laser scanning microscopy of biofilms formed by various combinations of colour-coded P. aeruginosa wild type and motility mutants. We show that mushroom-shaped multicellular structures in P. aeruginosa biofilms can form in a sequential process involving a non-motile bacterial subpopulation and a migrating bacterial subpopulation. The non-motile bacteria form the mushroom stalks by growth in certain foci of the biofilm. The migrating bacteria form the mushroom caps by climbing the stalks and aggregating on the tops in a process which is driven by type-IV pili. These results lead to a new model for biofilm formation by P. aeruginosa.

Biofilms↗

N-acetyl-beta-D-glucopyranosylamine: a potent T-state inhibitor of glycogen phosphorylase. A comparison with alpha-D-glucose.

Structure-based drug design has led to the discovery of a number of glucose analogue inhibitors of glycogen phosphorylase that have an increased affinity compared to alpha-D-glucose (Ki = 1.7 mM). The best inhibitor in the class of N-acyl derivatives of beta-D-glucopyranosylamine, N-acetyl-beta-D-glucopyranosylamine (1-GlcNAc), has been characterized by kinetic, ultracentrifugation, and crystallographic studies. 1-GlcNAc acts as a competitive inhibitor for both the b (Ki = 32 microM) and the a (Ki = 35 microM) forms of the enzyme with respect to glucose 1-phosphate and in synergism with caffeine, mimicking the binding of glucose. Sedimentation velocity experiments demonstrated that 1-GlcNAc was able to induce dissociation of tetrameric phosphorylase a and stabilization of the dimeric T-state conformation. Co-crystals of the phosphorylase b-1-GlcNAc-IMP complex were grown in space group P4(3)2(1)2, with native-like unit cell dimensions, and the complex structure has been refined to give a crystallographic R factor of 18.1%, for data between 8 and 2.3 A resolution. 1-GlcNAc binds tightly at the catalytic site of T-state phosphorylase b at approximately the same position as that of alpha-D-glucose. The ligand can be accommodated in the catalytic site with very little change in the protein structure and stabilizes the T-state conformation of the 280s loop by making several favorable contacts to Asn 284 of this loop. Structural comparisons show that the T-state phosphorylase b-1-GlcNAc-IMP complex structure is overall similar to the T-state phosphorylase b-alpha-D-glucose complex structure. The structure of the 1-GlcNAc complex provides a rational for the biochemical properties of the inhibitor.

Animals↗

Morphology and function of male genitalia (spermatophores) in Euscorpius italicus (Euscorpiidae, Scorpiones): complex spermatophore structures enable safe sperm transfer.

The structure and function of the spermatophore of Euscorpius italicus are analyzed. We show how the spermatophore gets shaped from two hemispermatophores and for the first time the sperm transfer mechanism is shown in detail, illustrating function and importance of all complex lobe structures of an euscorpiid spermatophore. A detailed description of the interaction of spermatophore and female genitalia is given. The capsular region of the spermatophore bears different lobes: The distal and basal lobes hook into two cavities on the inner side of the female's genital operculum. A so-called "crown-like structure" hooks into a membranous area in the genital atrium. During sperm transfer, these crown-like structures move backwards, in this way widening the female's genital opening. The sperm duct of the spermatophore is coated with numerous spicules on its outer side, which could serve as a sealing mechanism and/or may stimulate the female. Furthermore, we conclude that "safeguarding of sperm transfer" is one driving force for evolution of male genital complexity in scorpions, but also sexual selection by cryptic female choice could partly play a role.

Animals↗