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Selected AbstractsSynthesis and Structural Characterisation of Palladium and Group-12 Metal Complexes with a Hybrid Phosphanylphosphonate Ferrocene LigandEUROPEAN JOURNAL OF INORGANIC CHEMISTRY, Issue 5 2006Abstract Diethyl [1,-(diphenylphosphanyl)ferrocenyl]phosphonate (1) was synthesised by stepwise metallation/functionalisation of 1,1,-dibromoferrocene and studied as a ligand for palladium(II) and group-12 metals. Treatment of [PdCl2(cod)] (cod = ,2:,2 -cycloocta-1,5-diene) with 1 in 1:1 or 1:2 molar ratios gave, respectively, the dinuclear, chloride-bridged complex [{Pd(,-Cl)Cl(1 -,P2)}2] (2) and the mononuclear complex trans -[PdCl2(1 -,P2)2] (3), where 1 coordinates exclusively through the phosphane function. The reactions between 1 and group-12 metal bromides MBr2 in a 1:1 molar ratio gave the adducts [MBr2(1)] [M = Zn (4), Cd (5), and Hg (6)], whose crystal structures change considerably with the metal ion. Thus, whereas 4 is a molecular complex with 1 coordinating as an O1,P2 -chelate, its cadmium(II) analogue is a polymer built up from symmetric {CdBr(,-Br)}2 units interconnected by pairs of O1,P2 -bridging phosphanylphosphonate ligands. Finally, the mercury(II) complex 6 is a halide-bridged dimer, [{Hg(,-Br)Br(1 -,P2)}2]. However, this compound is structurally fluxional in solution (NMR spectra) and, in the crystal, it attains a structure similar to 5 owing to weak interactions between mercury and phosphonate-O1 atoms from adjacent molecules. An isomer to 6, [{HgBr2(1 -,2O1,P2)}2] (7), was isolated from attempted alkylation of 6 and structurally characterised as a dimer, where ligands 1 bridge two {HgBr2} units. All compounds were studied by spectroscopic methods (IR, NMR, mass) and the solid-state structures of 1, 2·,H2O, 3·4,CHCl3, 4, 5, 6·5,C6H6, and 7 have been determined by single-crystal X-ray diffraction. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2006) [source] Structure, regulation and evolution of Nox-family NADPH oxidases that produce reactive oxygen speciesFEBS JOURNAL, Issue 13 2008Hideki Sumimoto NADPH oxidases of the Nox family exist in various supergroups of eukaryotes but not in prokaryotes, and play crucial roles in a variety of biological processes, such as host defense, signal transduction, and hormone synthesis. In conjunction with NADPH oxidation, Nox enzymes reduce molecular oxygen to superoxide as a primary product, and this is further converted to various reactive oxygen species. The electron-transferring system in Nox is composed of the C-terminal cytoplasmic region homologous to the prokaryotic (and organelle) enzyme ferredoxin reductase and the N-terminal six transmembrane segments containing two hemes, a structure similar to that of cytochrome b of the mitochondrial bc1 complex. During the course of eukaryote evolution, Nox enzymes have developed regulatory mechanisms, depending on their functions, by inserting a regulatory domain (or motif) into their own sequences or by obtaining a tightly associated protein as a regulatory subunit. For example, one to four Ca2+ -binding EF-hand motifs are present at the N-termini in several subfamilies, such as the respiratory burst oxidase homolog (Rboh) subfamily in land plants (the supergroup Plantae), the NoxC subfamily in social amoebae (the Amoebozoa), and the Nox5 and dual oxidase (Duox) subfamilies in animals (the Opisthokonta), whereas an SH3 domain is inserted into the ferredoxin,NADP+ reductase region of two Nox enzymes in Naegleria gruberi, a unicellular organism that belongs to the supergroup Excavata. Members of the Nox1,4 subfamily in animals form a stable heterodimer with the membrane protein p22phox, which functions as a docking site for the SH3 domain-containing regulatory proteins p47phox, p67phox, and p40phox; the small GTPase Rac binds to p67phox (or its homologous protein), which serves as a switch for Nox activation. Similarly, Rac activates the fungal NoxA via binding to the p67phox -like protein Nox regulator (NoxR). In plants, on the other hand, this GTPase directly interacts with the N-terminus of Rboh, leading to superoxide production. Here I describe the regulation of Nox-family oxidases on the basis of three-dimensional structures and evolutionary conservation. [source] Isolation of flagellated bacteria implicated in Crohn's diseaseINFLAMMATORY BOWEL DISEASES, Issue 10 2007L. Wayne Duck BS Abstract Background: Serologic expression cloning has identified flagellins of the intestinal microbiota as immunodominant antigens in experimental colitis in mice and in individuals with Crohn's disease (CD). The present study was done to identify the microbial source of such flagellins. Methods: Using a variety of isolation and culture approaches, a number of previously unknown flagellated bacteria were isolated. Based on 16S ribosomal DNA sequences, these bacteria fall into the family Lachnospiraceae of the phylum Firmicutes. Results: Serum IgG from patients with CD and from mice with colitis reacted to the flagellins of these bacteria, and only their flagellins, whereas serum IgG from controls did not. The sequence of these flagellins demonstrate conserved amino- and carboxy-terminal domains that cluster phylogenetically and have a predicted 3D structure similar to Salmonella fliC, including an intact TLR5 binding site. The flagellin of 1 of these bacteria was likely O -glycosylated. Conclusions: The conserved immune response in both mouse and human to these previously unknown flagellins of the microbiota indicate that they play an important role in host,microbe interactions in the intestine. (Inflamm Bowel Dis 2007) [source] Molecular cloning, characterization, expression pattern and cellular distribution of an ovarian lipophorin receptor in the cockroach, Leucophaea maderaeINSECT MOLECULAR BIOLOGY, Issue 3 2009M. Tufail Abstract A cDNA that encodes a lipophorin receptor (LpR) with a predicted structure similar to that of the low density lipoprotein receptor (LDLR) gene superfamily was cloned from ovaries of the cockroach, Leucophaea maderae (Lem) and characterized. This is the first LpR sequenced from the order Dictyoptera. The cDNA has a length of 3362 bp coding for an 888-residue mature protein with a predicted molecular mass of ~99.14 kDa and a pI value of 4.68. The deduced amino acid sequence showed that the LemLpR harbours eight ligand-binding repeats (LBRs) at the N-terminus similar to the other insect LpRs, and thus resembles vertebrate VLDLRs. In addition to eight tandemly arranged LBRs, the five-domain receptor contains an O -linked sugar region and the classic LDLR internalization signal, FDNPVY. Northern blot analysis revealed the presence of ~4.0 kb ovarian mRNA that was transcribed throughout oogenesis with its peak especially during late previtellogenic and vitellogenic periods (from days 3 to 11). LpR transcript(s) or homologues of LDLRs were also detected in the head, midgut, Malpighian tubules, muscles and in the fat body. RNA in situ hybridization and immunocytochemistry localized the LpR mRNA and protein to germ line-derived cells, the oocytes, and revealed that LpR gene transcription and translation starts very early during oocyte differentiation in the germarium. LpR protein was evenly distributed throughout the cytoplasm during previtellogenic periods of oogenesis. However, during vitellogenic stages, the receptor was accumulated mainly in the cortex of the oocyte. Immunoblot analysis probed an ovarian LpR protein of ~115 and 97 kDa under reducing and nonreducing conditions, respectively. The protein signal appeared on day 2, increased every day and was high during vitellogenic periods from day 4 to day 7. Southern blot analysis suggested the presence of a single copy of the LpR gene in the genome of Le. maderae. [source] Displacement-controlled method and its applications to material non-linearityINTERNATIONAL JOURNAL FOR NUMERICAL AND ANALYTICAL METHODS IN GEOMECHANICS, Issue 3 2005H. Zheng Abstract For the analysis of non-linear problems, the displacement-controlled method (DCM) has a more extensive application scope and more powerful abilities than the load-controlled method (LCM). However, difficulties of the DCM's procedure not amenable to most finite element implementations of the conventional LCM have restricted its applications in geomechanics. By means of Sherman,Morrison's theorem, the solution of DCM is improved. The improved procedure is characterized by high efficiency, good numerical stability and a programme structure similar to LCM. Two aspects of applications of DCM are illustrated. The first application is to compute the response of a structure under a given load level like the conventional finite element analysis. The second application is to trace the equilibrium path of a structure under a given load distribution type. A simple but effective algorithm is presented for automatically adjusting the step length in tracing the equilibrium path. Examples illustrate that the proposed procedures are suited for modelling complicated non-linear problems in geomechanics. Copyright © 2005 John Wiley & Sons, Ltd. [source] The biology of vernix caseosaINTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 5 2006S. B. Hoath Synopsis The biology and physical properties of the uniquely human skin cream ,vernix caseosa' are discussed. This material coats the foetal skin surface during the last trimester of gestation and provides multiple beneficial functions for the foetus and newborn infant. Vernix has a complex structure similar to stratum corneum but lacks lipid lamellae and is more plastic due to the absence of desmosomal constraints. In utero, vernix is made in part by foetal sebaceous glands, interacts with pulmonary surfactant, detaches into the amniotic fluid, and is swallowed by the foetus. At the time of birth, vernix has a remarkably constant water content approximating 80%. Postnatally, vernix is simultaneously a cleanser, a moisturizer, an anti-infective, and an anti-oxidant. Vernix facilitates acid mantle development and supports normal bacterial colonization. Its hydrated cellular structure and unusual lipid composition provide a ,best' solution for the needs of the foetus and newborn, not least of which is the attraction of caregivers. Vernix is an important natural biomaterial of potential interest to cosmetic scientists, and other disciplines involved in product development and therapies targeting the complex interface between the stratum corneum and a changing terrestrial environment. Résumé La biologie et les propriétés physiques de la crème de peau exclusivement humaine ,Vernix caseosa « sont discutées. Ce matériau couvre la surface de la peau foetale pendant le dernier trimestre de gestation et remplit des fonctions avantageuses multiples pour le foetus et le nouveau-né. Le Vernix a une structure complexe semblable au stratum corneum, mais manque de lamelles lipidiques et est plus plastique en raison de l'absence de contraintes desmosomales. In utero, le Vernix est constitué en partie par des glandes sébacées foetales, il interagit avec le surfactant pulmonaire, il se détache dans le liquide amniotique et est avalé par le foetus. Au moment de la naissance, le Vernix a une teneur remarquablement constante en eau de l'ordre de 80%. Après la naissance, le Vernix devient simultanément un produit de lavage, un produit hydratant, un anti-infectieux et un anti-oxydant. Le Vernix facilite le développement du manteau acide et soutient la colonisation bactérienne normale. Sa structure cellulaire hydratée et sa composition en lipide inhabituelle en font ,une des meilleures » solutions pour les besoins du foetus et du nouveau-né, à laquelle le personnel soignant n'attache pas la moindre importance. Le Vernix est un biomatériau naturel important potentiellement intéressant pour les scientifiques cosméticiens et pour les autres disciplines impliquées dans le développement de produits et de thérapies visant l'interface complexe entre le stratum corneum et un environnement terrestre changeant. [source] Age-related changes in human meniscal glycosaminoglycansINTERNATIONAL JOURNAL OF EXPERIMENTAL PATHOLOGY, Issue 4 2004Gareth Blackburn Introduction With an increased human lifespan, a major challenge is now to ensure a concomitant increase in healthspan. Meniscal damage and degradation are common and are strongly correlated with subsequent osteoarthritis. Indeed, meniscal damage has been identified in about 60% of people over 60. Markers of pathology will facilitate intervention but first require normal age-related changes to be established. Methods Undamaged vascular and avascular regions of medial and lateral human menisci were comminuted and the tissue extracted into 4- m GuHCl and subject to associative CsCl density gradient centrifugation. Aggrecan and the small leucine rich PGs (SLRPs) were isolated and their GAG profiles examined by HPAEC fingerprinting, following enzyme depolymerization, and by an NMR spectroscopy. Results and discussion Analysis of aggrecan and the SLRPs show that there is a complex and dynamic pattern of KS, CS and DS abundance and distribution within human menisci, which changes with age. The abundance of SLRPs is higher in the avascular than vascular tissues, however, this is not reflected in the abundance of aggrecan which is present at similar levels in both tissue regions. The data show no other significant differences between medial and lateral and between vascular and avascular tissue regions. Analysis of the sulfation pattern of CS following digestion by ACII lyase, shows that in both aggrecan and SLRPs the 4-sulfation level falls with age from 20 to 35% in young tissues to 10,20% in older. Subsequent analyses following ABC lyase depolymerization, to include DS, shows very significant change with age from CS + DS 4-sulfation levels of ca. 40,55% in young tissue to ca. 15,30% in older. The difference between these datasets represents the contribution made by 4-sulfated DS. Thus, analysis of the difference suggests that DS makes a decreasing contribution to the CS/DS profile with age. Indeed, this is confirmed by an NMR analysis of these samples. Analysis of the resonances in the region 1.95,2.2 p.p.m. (ref to TSP) allows the estimation of the contribution made by DS, CS and KS. These data show that, in aggrecan, the contribution made by DS chains falls from ca. 10% in younger tissues to ca. 2,4% in older tissues. NMR analysis also shows that KS levels fall with age from ca. 15,20% in younger tissues to 5,10% in older tissues. Analysis of the structure of the KS chains shows chains with a structure similar to that of in articular cartilage but that at all ages there are very low levels of fucosylation (ca. 1,5%). Previous studies of age-related changes in CS/DS and KS structures have shown significant changes in the first 17 years of life, with only modest nonpathological changes after that time. These data from meniscal tissues do not show such a dramatic halting of normal age-related changes. Indeed, the data show gradual age-related changes in DS, CS and KS abundance and structure throughout life. These baseline age-related changes data will now allow the analysis of pathology-related changes. [source] Perceived adverse patient outcomes correlated to nurses' workload in medical and surgical wards of selected hospitals in KuwaitJOURNAL OF CLINICAL NURSING, Issue 4 2009Fatimah Al-Kandari Aim., This study was carried out to identify the perceived adverse patient outcomes as related to nurses' workload. It also assessed nurses' perception of variables contributing to the workload and adverse patient outcomes. Background., Several studies have been published on adverse patient outcomes in which a correlation was found between nurses' workload and some adverse patient outcomes. Design., A cross-sectional survey was conducted between registered nurses (n = 780) working in medical and surgical wards of five general governmental hospitals in Kuwait. Data collection instruments., Data were collected using a self-administered questionnaire consisting of three sections to elicit information about the sample characteristics, perception of workload and perceived adverse patient outcomes during the last shift and last working week. Results., The three major perceived adverse outcomes reported by the nurses while on duty during their last shift were: complaints from patients and families (2%), patients received a late dose or missed a dose of medication (1·8%) and occurrences of pressure ulcer (1·5%). Similarly, the reported adverse outcomes over the past week were complaints from patients and families (5%), patients received a late dose or missed a dose of medication (5·3%) and discovery of a urinary tract infection (3·7%). Increases in nurse-patient load, bed occupancy rate, unstable patients' condition, extra ordinary life support efforts and non-nursing tasks; all correlated positively with perceived adverse patient outcomes. Conclusion., This study sheds light on an important issue affecting patient safety and quality of care as perceived by the nurses themselves as caregivers. Relevance to clinical practice., Nurses' perception of variables contributing to adverse patient outcomes and their workload could significantly affect the provided nursing care and nursing care recipients. The findings could help in policy formulation and planning strategies to decrease adverse patient outcomes in many countries with a health care structure similar to that of Kuwait. [source] Supramolecular inclusion complexes of star-shaped poly(,-caprolactone) with ,-cyclodextrinJOURNAL OF POLYMER SCIENCE (IN TWO SECTIONS), Issue 20 2005Lu Wang Abstract Both star-shaped poly(,-caprolactone) (PCL) having 4 arms (4sPCL) and 6 arms (6sPCL) and linear PCL having 1 arm (LPCL) and 2 arms (2LPCL) were synthesized and then investigated for inclusion complexation with ,-cyclodextrin (,-CD). The supramolecular inclusion complexes (ICs) were in detail characterized by 1H NMR, differential scanning calorimetry, thermogravimetric analysis, wide angle X-ray diffraction, solid-state carbon nuclear magnetic resonance spectroscopy using cross-polarization and magic-angle spinning, and Fourier transform infrared, respectively. The stoichiometry (CL:CD, mol:mol) of all ICs increased with the increasing branch arm of PCL polymers, and it was in the order of ,-CD-6sPCL1 ICs > ,-CD-4sPCL ICs > ,-CD-2LPCL ICs > ,-CD-LPCL ICs. All analyses indicated that the branch arms of star-shaped PCL polymers were included into the hydrophobic ,-CD cavities and their original crystalline properties were completely suppressed. Moreover, the ICs of star-shaped PCL with ,-CD had a channel-type crystalline structure similar to that formed between the linear PCL and ,-CD. Furthermore, the thermal stability of the free PCL polymers probably controlled that of the guest polymers included in the ICs. © 2005 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 43: 4721,4730, 2005 [source] Software Maintenance Maturity Model (SMmm): the software maintenance process modelJOURNAL OF SOFTWARE MAINTENANCE AND EVOLUTION: RESEARCH AND PRACTICE, Issue 3 2005Alain April Abstract We address the assessment and improvement of the software maintenance function by proposing a maturity model for daily software maintenance activities: the Software Maintenance Maturity Model (SMmm). The software maintenance function suffers from a scarcity of management models to facilitate its evaluation, management, and continuous improvement. The SMmm addresses the unique activities of software maintenance while preserving a structure similar to that of the Capability Maturity Model integration (CMMi). It is designed to be used as a complement to that model. The SMmm is based on practitioners' experience, international standards, and the seminal literature on software maintenance. We present the model's purpose, scope, foundation, and architecture, followed by its initial validation. Copyright © 2005 John Wiley & Sons, Ltd. [source] Comparative observations on the cyst shells of seven Artemia strains from ChinaMICROSCOPY RESEARCH AND TECHNIQUE, Issue 8 2007Sufeng Wang Abstract The quiescent Artemia cysts of seven geographical origins in China were examined with scanning and transmission electron microscopes. SEM observations on cysts of these Artemia strains showed that the surface topography of cyst shells could be categorized into 6 types: complete smooth surface; smooth surface with sparsely distributed glabrate humps; surface with densely arranged wart-like humps that are composed of packed minute tubercles; rugged surface, with densely arranged tubercles not piling up to form larger humps; shallow-pocked surface; and surface with numerous and densely spaced pore-like fossulae. Some of the patterns were strain specific [e.g., cysts from Ga Hai (GH) are characterized by having a surface with wart-like ornaments that are composed of packed minute tubercles, rugged surface is only found in Chengkou (CK) cysts], and apparent intrastrain variation of cyst surface topography was found in Xizang (XZ), Jingyu Hu, and Xie Chi (SIN) strains. TEM studies on the ultrastructure of cyst shells revealed an apparent divergence in the structure of outer cuticular membrane (OCM) among Artemia strains. In CK, Aqqikkol Hu (AQK), SIN, and GH strains, it is a normal, asymmetrical, and multi-layered structure similar to those described in previous works. In XZ, JYH, and Lagkor Co (LGC) strains, however, the OCM is not obviously multi-layered and the borderlines between OCM and adjacent layers seem indistinct. The present results suggest that the diversity of the surface topography of Artemia cysts may be an available tool for identifying certain Artemia strains as well as for tracking the origins of some Artemia cysts, and the hypoplastic OCM may be a characteristic of the species A. tibetiana. Microsc. Res. Tech., 2007. © 2007 Wiley-Liss, Inc. [source] AMMONOID SHELL STRUCTURES OF PRIMARY ORGANIC COMPOSITIONPALAEONTOLOGY, Issue 6 2007CHRISTIAN KLUG Abstract:, Palaeozoic and Mesozoic cephalopod conchs occasionally reveal dark organic coatings at the aperture. A number of these coatings, including still unrecorded examples, are described, figured and interpreted herein. On the basis of elemental analysis, actualistic comparison and a comparison with Triassic bivalves, some of these coatings are shown to consist of apatite and primarily probably of conchiolin (and also probably melanin). In several Mesozoic ammonoid genera such as Paranannites, Psiloceras, Lytoceras, Phylloceras, Harpoceras and Chondroceras, some of these coatings (recorded herein for most of these taxa for the first time) are interpreted as a structure similar to the black band, which was previously known only from Recent Allonautilus and Nautilus. In contrast to these nautilid genera, however, the organic material of some Mesozoic ammonoids was not deposited on the inside of the shell but externally, albeit positioned at the terminal aperture as in Recent nautilids. Some ammonoids of Carboniferous and Triassic age show several such bands at more or less regular angular distances on the ultimate whorls and at the aperture, e.g. Nomismoceras, Gatherites, Owenites, Paranannites, Juvenites and Melagathiceratidae gen. et sp. nov. Triassic material from Oman shows that the black coating was probably secreted from the inside, because the position of this organic deposit changes from interior to exterior in an anterior direction (i.e. adaperturally). This structure has previously been referred to as a ,false colour pattern' and is here interpreted as having been formed at an interim aperture or megastria (,alter Mundrand'). All structures discussed in the paper are considered to have been secreted by a single organ and to have been initiated by some form of stress or adverse conditions. Thus, certain environmental parameters and growth anomalies appear to have influenced their formation. [source] A Role for Internal Water Molecules in Proton Affinity Changes in the Schiff Base and Asp85 for One-way Proton Transfer in Bacteriorhodopsin,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 4 2008Joel E. Morgan Light-induced proton pumping in bacteriorhodospin is carried out through five proton transfer steps. We propose that the proton transfer to Asp85 from the Schiff base in the L-to-M transition is accompanied by the relocation of a water cluster on the cytoplasmic side of the Schiff base from a site close to the Schiff base in L to the Phe219-Thr46 region in M. The water cluster present in L, formed at 170 K, is more rigid than that at room temperature. This may be responsible for blocking the conversion of L to M at 170 K. In the photocycle at room temperature, this water cluster returns to the site close to the Schiff base in N, with a rigid structure similar to that of L at 170 K. The increase in the proton affinity of Asp85, which is a prerequisite for the one-way proton transfer in the M-to-N transition, is suggested to be facilitated by a structural change which disrupts interactions between Asp212 and the Schiff base, and between Asp212 and Arg82. We propose that this liberation of Asp212 is accompanied by a rearrangement of the structure of water molecules between Asp85 and Asp212, stabilizing the protonated Asp85 in M. [source] A uniquely high number of ftsZ genes in the moss Physcomitrella patensPLANT BIOLOGY, Issue 5 2009A. Martin Abstract Plant FtsZ proteins are encoded by two small nuclear gene families (FtsZ1 and FtsZ2) and are involved in chloroplast division. From the moss Physcomitrella patens, four FtsZ proteins, two in each nuclear gene family, have been characterised and described so far. In the recently sequenced P. patens genome, we have now found a fifth ftsZ gene. This novel gene has a genomic structure similar to PpftsZ1-1. According to phylogenetic analysis, the encoded protein is a member of the FtsZ1 family, while PpFtsZ1-2, together with an orthologue from Selaginella moellendorffii, forms a separate clade. Further, this new gene is expressed in different gametophytic tissues and the encoded protein forms filamentous networks in chloroplasts, is found in stromules, and acts in plastid division. Based on all these results, we have renamed the PpFtsZ proteins of family 1 and suggest the existence of a third FtsZ family. No species is known to encode more FtsZ proteins per haploid genome than P. patens. [source] Contributions of hydrophobic domain interface interactions to the folding and stability of human ,D-crystallinPROTEIN SCIENCE, Issue 3 2005Shannon L. Flaugh Abstract Human ,D-crystallin (H,D-Crys) is a monomeric eye lens protein composed of two highly homologous ,-sheet domains. The domains interact through interdomain side chain contacts forming two structurally distinct regions, a central hydrophobic cluster and peripheral residues. The hydrophobic cluster contains Met43, Phe56, and Ile81 from the N-terminal domain (N-td) and Val132, Leu145, and Val170 from the C-terminal domain (C-td). Equilibrium unfolding/refolding of wild-type H,D-Crys in guanidine hydrochloride (GuHCl) was best fit to a three-state model with transition midpoints of 2.2 and 2.8 M GuHCl. The two transitions likely corresponded to sequential unfolding/refolding of the N-td and the C-td. Previous kinetic experiments revealed that the C-td refolds more rapidly than the N-td. We constructed alanine substitutions of the hydrophobic interface residues to analyze their roles in folding and stability. After purification from E. coli, all mutant proteins adopted a native-like structure similar to wild type. The mutants F56A, I81A, V132A, and L145A had a destabilized N-td, causing greater population of the single folded domain intermediate. Compared to wild type, these mutants also had reduced rates for productive refolding of the N-td but not the C-td. These data suggest a refolding pathway where the domain interface residues of the refolded C-td act as a nucleating center for refolding of the N-td. Specificity of domain interface interactions is likely important for preventing incorrect associations in the high protein concentrations of the lens nucleus. [source] Transmembrane signal transduction of the ,IIb,3 integrinPROTEIN SCIENCE, Issue 7 2002Kay E. Gottschalk Abstract Integrins are composed of noncovalently bound dimers of an ,- and a ,-subunit. They play an important role in cell-matrix adhesion and signal transduction through the cell membrane. Signal transduction can be initiated by the binding of intracellular proteins to the integrin. Binding leads to a major conformational change. The change is passed on to the extracellular domain through the membrane. The affinity of the extracellular domain to certain ligands increases; thus at least two states exist, a low-affinity and a high-affinity state. The conformations and conformational changes of the transmembrane (TM) domain are the focus of our interest. We show by a global search of helix,helix interactions that the TM section of the family of integrins are capable of adopting a structure similar to the structure of the homodimeric TM protein Glycophorin A. For the ,IIb,3 integrin, this structural motif represents the high-affinity state. A second conformation of the TM domain of ,IIb,3 is identified as the low-affinity state by known mutational and nuclear magnetic resonance (NMR) studies. A transition between these two states was determined by molecular dynamics (MD) calculations. On the basis of these calculations, we propose a three-state mechanism. [source] Protein,protein docking with multiple residue conformations and residue substitutionsPROTEIN SCIENCE, Issue 6 2002David M. Lorber Abstract The protein docking problem has two major aspects: sampling conformations and orientations, and scoring them for fit. To investigate the extent to which the protein docking problem may be attributed to the sampling of ligand side-chain conformations, multiple conformations of multiple residues were calculated for the uncomplexed (unbound) structures of protein ligands. These ligand conformations were docked into both the complexed (bound) and unbound conformations of the cognate receptors, and their energies were evaluated using an atomistic potential function. The following questions were considered: (1) does the ensemble of precalculated ligand conformations contain a structure similar to the bound form of the ligand? (2) Can the large number of conformations that are calculated be efficiently docked into the receptors? (3) Can near-native complexes be distinguished from non-native complexes? Results from seven test systems suggest that the precalculated ensembles do include side-chain conformations similar to those adopted in the experimental complexes. By assuming additivity among the side chains, the ensemble can be docked in less than 12 h on a desktop computer. These multiconformer dockings produce near-native complexes and also non-native complexes. When docked against the bound conformations of the receptors, the near-native complexes of the unbound ligand were always distinguishable from the non-native complexes. When docked against the unbound conformations of the receptors, the near-native dockings could usually, but not always, be distinguished from the non-native complexes. In every case, docking the unbound ligands with flexible side chains led to better energies and a better distinction between near-native and non-native fits. An extension of this algorithm allowed for docking multiple residue substitutions (mutants) in addition to multiple conformations. The rankings of the docked mutant proteins correlated with experimental binding affinities. These results suggest that sampling multiple residue conformations and residue substitutions of the unbound ligand contributes to, but does not fully provide, a solution to the protein docking problem. Conformational sampling allows a classical atomistic scoring function to be used; such a function may contribute to better selectivity between near-native and non-native complexes. Allowing for receptor flexibility may further extend these results. [source] Design and analysis of industrial strip-plot experimentsQUALITY AND RELIABILITY ENGINEERING INTERNATIONAL, Issue 2 2010Heidi Arnouts Abstract The cost of experimentation can often be reduced by forgoing complete randomization. A well-known design with restricted randomization is a split-plot design, which is commonly used in industry when some experimental factors are harder to change than others or when a two-stage production process is studied. Split-plot designs are also often used in robust product design to develop products that are insensitive to environmental or noise factors. Another, lesser known, type of experimental design plan that can be used in such situations is the strip-plot experimental design. Strip-plot designs are economically attractive in situations where the factors are hard to change and the process under investigation consists of two distinct stages, and where it is possible to apply the second stage to groups of semi-finished products from the first stage. They have a correlation structure similar to row,column designs and can be seen as special cases of split-lot designs. In this paper, we show how optimal design of experiments allows for the creation of a broad range of strip-plot designs. Copyright © 2009 John Wiley & Sons, Ltd. [source] Mass spectra of nitro- ,,, -dihalostyrenesRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 9 2004Shaw-Tao Lin The electron ionization (EI) fragmentation of nitro- ,,, -dihalostyrenes depends strongly on the specific halogens present as well as on the position of the nitro group. ,,, -Difluorostyrenes yielded mainly fluoroacetylene ions as the base peaks and o -nitro- ,,, -dihalostyrene possibly furnished the ion with a structure similar to that of benzo[a]isoxazole (11). The structural identity of the ion was verified by comparing the collision-induced dissociation (CID) spectra of m/z 119 ions from p - and o -nitro- ,,, -difluorostyrene, o -nitro- ,,, -dibromostyrene and compound 11. Copyright © 2004 John Wiley & Sons, Ltd. [source] The Mucosa of the Digestive Tract in Micropogonias furnieri: A Light and Electron Microscope ApproachANATOMIA, HISTOLOGIA, EMBRYOLOGIA, Issue 4 2008A. O. Diaz Summary The histomorphological aspects as well as the histochemical content and distribution of glycoproteins (GPs) in the mucosa of the digestive tract of the white croaker Micropogonias furnieri were studied. The buccopharyngeal cavity and the esophagous showed a squamous stratified epithelium with mucous cells. The stomach presented three portions: cardias, fundus and pylorus. Tubular glands formed by a single type of gland cell were located along the cardias and fundus. Histochemical tests showed that the buccopharyngeal cavity and the esophagous presented the largest amount of the different types of mucosubstances. Both organs showed abundant secretory mucous cells that synthesize large quantities of neutral, sulphated and sialylated GPs. The surface epithelium in the cardias and fundus synthesized and secreted scarce sialylated and neutral GPs whereas the secretions of the apical surface were abundant. The pylorus secreted large amounts of neutral as well as sulphated and sialylated GPs. Gland cells secreted neutral GPs. The ultrastructural features of the gut cells were quite similar to those of other teleosts. The buccopharyngeal cavity and the esophagous surface epithelial cells, identified by their superficial localization, were characterized by cytoplasmic vesicles of different size. Abundant goblet cells with secretory mucous granules were also present. Gastric glands in the stomach contained just one form of cell with a fine structure similar to cells that secrete pepsinogen. [source] Laser-guided direct writing for three-dimensional tissue engineeringBIOTECHNOLOGY & BIOENGINEERING, Issue 2 2005Yaakov Nahmias Abstract One of the principal limitations to the size of an engineered tissue is oxygen and nutrient transport. Lacking a vascular bed, cells embedded in an engineered tissue will consume all available oxygen within hours while out branching blood vessels will take days to vascularize the implanted tissue. One possible solution is to directly write vascular structures within the engineered tissue prior to implantation, reconstructing the tissue according to its native architecture. The cell patterning technique, laser-guided direct writing (LGDW), can pattern multiple cells types with micrometer resolution on arbitrary surfaces, including biological gels. Here we show that LGDW can pattern human umbilical vein endothelial cells (HUVEC) in two- and three-dimensions with micrometer accuracy. By patterning HUVEC on Matrigel, we can direct their self-assembly into vascular structures along the desired pattern. Finally, co-culturing the vascular structures with hepatocytes resulted in an aggregated tubular structure similar in organization to a hepatic sinusoid. This capability can facilitate studies of tissue architecture at the single cell level, and of heterotypic interactions underlying processes such as liver and pancreas morphogenesis, differentiation, and angiogenesis. Copyright © 2005 Wiley Periodicals, Inc. [source] Intra-hepatic arterial administration with miriplatin suspended in an oily lymphographic agent inhibits the growth of human hepatoma cells orthotopically implanted in nude ratsCANCER SCIENCE, Issue 1 2009Mitsuharu Hanada Miriplatin is a lipophilic platinum complex which contains myristates as leaving groups and diaminocyclohexane as a carrier ligand. In order to examine in vivo the antitumor activities of miriplatin suspended in an oily lymphographic agent (Lipiodol Ultra-Fluide®, LPD) against human hepatocellular carcinoma (HCC) after the intra-hepatic arterial administration, we have developed a novel orthotopic model of HCC in which the human hepatoma cell line Li-7 was successively implanted and maintained in the liver of nude rats. Li-7 tumors established in nude rat livers displayed a trabecular structure similar to their original morphology, and were exclusively supplied by the hepatic artery, suggesting that they exhibited in part the conditions of human HCC. Miriplatin suspended in LPD (miriplatin/LPD) administered into the hepatic artery of this model dose-dependently inhibited the growth of Li-7 tumors without markedly enhancing body weight loss and caused a significant reduction in the growth rate at a dose of 400 µg/head compared to LPD alone. In addition, at the therapeutic dose, miriplatin/LPD as well as cisplatin suspended in LPD (400 µg/head) was shown to be more active than zinostatin stimalamer suspended in LPD (20 µg/head) against Li-7 tumors after a single intra-hepatic arterial administration. These results suggest miriplatin to be a suitable candidate for use in transarterial chemoembolization (Cancer Sci 2009; 100: 189,194) [source] Cloning a novel developmental regulating gene, Xotx5: Its potential role in anterior formation in Xenopus laevisDEVELOPMENT GROWTH & DIFFERENTIATION, Issue 2 2000Hiroki Kuroda The vertebrate Otx gene family is related to otd, a gene contributing to head development in Drosophila. In Xenopus, Xotx1, Xotx2, and Xotx4 have already been isolated and analyzed. Here the cloning, developmental expression and functions of the additional Otx Xenopus gene, Xotx5 are reported. This latter gene shows a greater degree of homology to Xotx2 than Xotx1 and Xotx4. Xotx5 was initially expressed in Spemann's organizer and later in the anterior region. Ectopic expression of Xotx5 had similar effects to other Xotx genes in impairing trunk and tail development, and especially similar effects to Xotx2 in causing secondary cement glands. Taken together, these findings suggest that Xotx5 stimulates the formation of the anterior regions and represses the formation of posterior structures similar to Xotx2. [source] Micronuclei and chromatid buds are the result of related genotoxic eventsENVIRONMENTAL AND MOLECULAR MUTAGENESIS, Issue 1 2001Luis Serrano-García Abstract Chromatin buds (CHB), broken eggs, or budding cell nuclei are structures similar to micronuclei (MN) in shape, structure, and size, which are linked to the main nuclei of cells by a thread or stalks of chromatin. They have been observed in numerous cell types and there are reports of their existence relating them with MN or with genotoxic events. However, there is no systematic study reporting their frequency and no experiment has been done to ascertain whether they are really induced by genotoxins. Furthermore, they have been discarded as genotoxic events with the argument that they are not formed in dividing cells. Studies are presented here that indicate that CHB can be considered as genotoxic events and that their origin is comparable to that of MN. Bromodeoxyuridine (BrdU) was used to label proliferating lymphocytes, which were later identified by means of an immunohistochemical method, using the H2O2,DAB stain. The results show that CHB are consistently formed where MN are seen. CHB were induced by the clastogen mitomycin C (MMC) as well as by the aneuploidogen colcemid, with frequencies similar to MN in both cases, and to multinucleated cells in the case of colcemid. CHB occur in lymphocytes of smokers with frequencies similar to those of MN, and we found that the infection with Taenia solium metacestodes induced a comparable increase of both MN and CHB frequency in lymphocytes from pigs. Environ. Mol. Mutagen. 38:38,45, 2001 © 2001 Wiley-Liss, Inc. [source] Modulation of the Lifetime of Water Bound to Lanthanide Metal Ions in Complexes with Ligands Derived from 1,4,7,10-Tetraazacyclododecane Tetraacetate (DOTA)HELVETICA CHIMICA ACTA, Issue 5 2005Shanrong Zhang A series of di- and tetraamide derivatives of DOTA were synthesized, and their lanthanide(III) complexes were examined by multinuclear 1H-, 13C-, and 17O-NMR spectroscopy, and compared with literature data of similar, known complexes (Table). All ligands formed structures similar to the parent [LnIII(DOTA)], complexes, with four N-atoms and four O-atoms from DOTA and one O-atom from the inner-sphere water molecules. Interestingly, the lifetimes ,M of the inner-sphere, metal-bound water molecules vary widely, ranging from nano- to milliseconds, depending on the identity of the pendent amide side chains. In general, positively charged [LnIII(DOTA-tetraamide)]3+ complexes display the longest residence times (high ,M values), while complexes with additional charged functional groups on the extended amides display much smaller ,M values, even when the side groups are not directly coordinated to the central Ln3+ ions. The design of novel [LnIII(DOTA-tetraamide)]3+ complexes with a wide, tunable range of ,M values is of prime importance for the application of fast-responding, paramagnetic chemical-exchange-saturation-transfer (PARACEST) imaging agents used for the study of physiological and metabolic processes. [source] Development, characterization, and validation of porous carbonated hydroxyapatite bone cementJOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 2 2009Pei-Fu Tang Abstract Carbonated hydroxyapatite (CHA) bone cement is capable of self-setting and forming structures similar to mineralized bone. Conventional CHA leaves little room for new bone formation and delays remodeling. The purposes of this study were to develop porous CHA (PCHA) bone cement and to investigate its physicochemical properties, biocompatibility, biodegradation, and in vivo bone repair potential. Vesicants were added to modify CHA, and the solidification time, porosity, and pore size of the PCHA cements were examined. The cytotoxicity and bone repair potential of PCHA were tested in a rabbit bone defect model and assessed by x-ray, histological examination, and mechanical testing. The porosity of the modified PCHA was 36%; 90.23% of the pores were greater than 70 ,m, with a calcium/phosphate ratio of 1.64 and a solidification time of 15 minutes. The PCHA did not affect bone cell growth in vitro, and the degrading time of the PCHA was two and four times faster in vitro and in vivo when compared to CHA. In the bone defect model, the amount of new bone formation in the PCHA-treated group was eight times greater than that of the CHA group; the compressive strength of the PCHA setting was relatively weak in the first weeks but increased significantly at 8 to 16 weeks compared to the CHA group. The PCHA has stable physicochemical properties and excellent biocompatibility; it degrades faster than CHA, provides more porous spaces for new bone ingrowths, and may be a new form of bone cement for the management of bone defects. © 2009 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 2009 [source] Conformation-activity relationships of cyclo -constrained µ/, opioid agonists derived from the N -terminal tetrapeptide segment of dermorphin/deltorphinJOURNAL OF PEPTIDE SCIENCE, Issue 8 2008Sylwia Rodziewicz-Motowid Abstract The N -terminal tetrapeptide segments of dermorphin (Tyr,D -Ala,Phe,Gly,Tyr,Pro,Ser,NH2) and deltorphin (Tyr,D -Ala,Phe,Asp/Glu,Val,Val,Gly,NH2) are agonists at the opioid receptors µ and ,, respectively. [D -Arg2, Lys4]-dermorphin-(1,4) amide (Tyr,D -Arg,Phe,Lys,NH2, DALDA) and [Dmt1]DALDA (where Dmt is 2,,6,-dimethyltyrosine) are among the most potent and selective µ-agonists reported to date, both in vitro (having picomolar µ receptor affinity) and in vivo. In this communication, conformation-activity studies of the following four cyclic analogs of DALDA are presented and discussed: the lead peptide S2,S4 -cyclo (Tyr,D -Cys,Phe,Cys,NH2), constrained by means of an S4.2S4.4 disulfide between Cys2 and Cys4; its two cis and transC4.2C4.4 -olefinic dicarba analogs, and the product of saturation of them both. They are potent nonselective or moderately µ-selective opioid agonists in vitro. They have been synthesized and tested earlier [Berezowska I, Chung NN, Lemieux C, Wilkes BC, and Schiller PW, Acta Biochim Polon 53, 2006, 73,76]. We have studied their conformations using NMR and molecular dynamics. With major conformational constraints imposed by the 11-membered ring spanning residues 2,4, they show well defined conformations of this ring, while the exocylic Tyr1 and Phe3 side chains still have significant conformational freedom. The more active and selective µ versus , disulfide and saturated dicarba agonists seem to have in common: (i) their ring structures more flexilble than those of the other two and (ii) their ring structures similar to each other and more diverse than those in the other two. Given this and the small size of the peptides having confirmed bioactivity profiles, there is a chance that their conformations determined in solution approach receptor-bound conformations. Copyright © 2008 European Peptide Society and John Wiley & Sons, Ltd. [source] Synthesis of Fluorinated Chitin Derivatives via Enzymatic PolymerizationMACROMOLECULAR BIOSCIENCE, Issue 10 2006Akira Makino Abstract Summary: Synthesis of fluorinated chitin derivatives has been achieved using chitinase from Bacillus sp. as a catalyst. 6,-Fluoro- (1a), 6-fluoro- (1b) and 6,6,-difluoro- (1c) chitobiose oxazoline derivatives were newly prepared as TSAS monomers for chitinase. Ring-opening polyaddition of these monomers proceeded effectively at pH 8.0,9.0 and 30,40,°C, giving rise to alternatingly 6-fluorinated chitin derivatives (2a and 2b) from 1a and 1b, and fully 6-fluorinated chitin derivative (2c) from 1c under total control of regioselectivity and stereochemistry. XRD measurements revealed that polysaccharides 2a and 2b had crystalline structures similar to that of , -chitin. [source] The first appearance of the red sequence of galaxies in proto-clusters at 2 ,z, 3MONTHLY NOTICES OF THE ROYAL ASTRONOMICAL SOCIETY, Issue 4 2007Tadayuki Kodama ABSTRACT We explore the evolved galaxy population in the proto-clusters around four high- z radio galaxies at 2 ,z, 3 based on wide-field near-infrared (NIR) imaging. Three of the four fields are known proto-clusters as demonstrated by overdensities of line-emitting galaxies at the same redshifts as the radio galaxies found by narrow-band surveys and spectroscopic follow-up observations. We imaged the fields of three targets (PKS 1138,262, USS 0943,242 and MRC 0316,257) to a depth of Ks, 22 (Vega magnitude, 5,) over a 4 × 7 arcmin2 area centred on the radio galaxies with a new wide-field NIR camera, Multi-Object Infra-Red Camera and Spectrograph (MOIRCS), on the Subaru Telescope. Another target (USS 1558,003) was observed with Son of ISAAC on the New technology Telescope (NTT) to a depth of Ks= 20.5 (5,) over a 5 × 5 arcmin2 area. We apply colour cuts in J,Ks and/or JHKs in order to exclusively search for galaxies located at high redshifts: z > 2. To the 5, limiting magnitudes, we see a significant excess of NIR-selected galaxies by a factor of 2 to 3 compared to those found in the field of GOODS-South. The spatial distribution of these NIR-selected galaxies is not uniform and traces structures similar to those of emission-line galaxies, although the samples of NIR-selected galaxies and emitters show little overlap, from which we conclude that the former tend to be an evolved population with much higher stellar mass than the latter, young and active emitters. We focus on the NIR colour,magnitude sequence of the evolved population and find that the bright-end (Mstars > 1011 M,) of the red sequence is well populated by z, 2 but much less so in the z, 3 proto-clusters. This may imply that the bright-end of the colour,magnitude sequence first appeared between z= 3 and 2, an era coinciding with the appearance of sub-mm galaxies and the peak of the cosmic star formation rate. Our observations show that during the same epoch, massive galaxies are forming in high-density environments by vigorous star formation and assembly. [source] Mixed epithelial and stromal tumor of the kidney in a 12-year-old girlPATHOLOGY INTERNATIONAL, Issue 10 2005Noboru Hara Mixed epithelial and stromal tumor of the kidney (MESTK) is a rare kidney neoplasm that almost exclusively occurs in perimenopausal women, and long-term estrogen replacement is relevant to its pathogenesis. Herein is described an atypical case of MESTK uncovered in a 12-year-old premenarcheal girl without a history of prior estrogen use. On surgical specimen it was found that the well-circumscribed tumor measuring 14 cm arose from the lower pole of the right kidney, showing solid and fibrous-cystic areas. Microscopically, it was composed both of epithelial structures similar to renal tubules and stroma comprising non-specific spindle cells. Some intratumoral tubules showed affinities to distal-nephron-specific lectins, and those immunoreactive for proximal-tubule-specific CD15 were also present. In addition, primitive ductal structures were reactive both for CD15 and lectins, but immature epithelial elements typical of nephroblastoma were absent. Spindle cells were positive for actin, desmin and vimentin, and expressed progesterone and estrogen receptors. The tumor was comparable with MESTK, although some epithelia were associated with the immunophenotype of proximal tubules. The patient was free of disease postoperatively for 40 months. In the present case, remnants of the primitive periductal mesenchyme might be promoted to neoplastic cells by a sex-steroid surge during puberty. [source] |