PMID- 19216472 TI - [Enteric duplication: unusual localisation of ciliated epithelium]. PMID- 19216473 TI - [Intrascrotal adenomatoid tumors]. PMID- 19216474 TI - [Hepatic undifferentiated (embryonal) sarcoma arising in a mesenchymal hamartoma: association or malignant transformation]. PMID- 19216478 TI - [Value and limits of a temporary admission to an Alzheimer care unit]. PMID- 19216479 TI - [Coordinating nurse in a home care service. Interview by Sarah Trotet]. PMID- 19216480 TI - [Home care services today, a rich but complex sector]. PMID- 19216481 TI - [The coordination of actors, indispensable links in home care]. PMID- 19216482 TI - [The collaboration work of health and social sectors, one of the challenges of home care]. PMID- 19216483 TI - [Returning home after a hospitalization, a complex organization]. PMID- 19216484 TI - [Resorting to home care help, many possibilities for the particular needs]. PMID- 19216485 TI - [Review of the home care services regulation field]. PMID- 19216486 TI - [The development of a personalized nursing care plan for a retired person]. PMID- 19216487 TI - [The community health nurse, a pivotal actor in home care]. PMID- 19216488 TI - [Health care networks, an innovative apparatus for coordination of home care services]. PMID- 19216489 TI - [7/12--The coordinating physician]. PMID- 19216490 TI - [Nurses, patients and families. 4/8--The role of the family]. PMID- 19216491 TI - [1/6--Cataracts]. PMID- 19216492 TI - Covalent modification of lysine residues by allyl isothiocyanate in physiological conditions: plausible transformation of isothiocyanate from thiol to amine. AB - We investigated the reactivity of allyl isothiocyanate (AITC) with amino groups under physiological conditions. First, the chemical reaction of AITC with bovine serum albumin (BSA) was investigated. When BSA was incubated with AITC in a phosphate buffer (pH 7.4), the loss of Lys residues was observed. Second, the Lys residue N(alpha)-benzoyl-glycyl-L-lysine (BGK) was reacted with AITC in the buffer, and a novel peak was detected using high performance liquid chromatography (HPLC). The peak was purified and identified as AITC-modified BGK with a N(epsilon)-thiocarbamoyl linkage. However, a thiol residue is known to be a predominant target of an isothiocyanate (ITC). Although AITC may react with a thiol moiety in vivo, a thiocarbamoyl linkage between ITC and thiol is unstable, and an AITC molecule may be regenerated. To prove the plausible transformation of ITC from thiol to amine, synthetic AITC-conjugated N(alpha)-acetyl-L-cysteine (NAC) was incubated with BGK at 37 degrees C in physiological buffer, and the generation of AITC-Lys was analyzed. The loss of the AITC-NAC adduct corresponded to the formation of the AITC-BGK adduct. Furthermore, using a novel monoclonal antibody (A4C7mAb) specific for AITC-Lys, we found that the AITC-Lys residue was generated from the reaction between AITC-NAC and BSA. Although AITC preferentially reacts with thiol rather than with Lys, AITC can be liberated from thiols and can then react with amino groups. The ITC-Lys adduct may be a useful marker for ITC target molecules. PMID- 19216494 TI - Hierarchal contribution of N- and C-terminal sequences to the differential localization of homologous sodium-dependent vitamin C transporters, SVCT1 and SVCT2, in epithelial cells. AB - Human sodium-dependent vitamin C transporters, SVCT1 and SVCT2, share 66% sequence identity yet localize in the apical and basolateral membranes of epithelial cells, respectively. This pair thus serves as a model for studying multipass membrane protein targeting. Domain swaps, deletions, insertions, and point mutations were performed on EGFP-tagged hSVCT1 and hSVCT2 plasmids. Mutant proteins stably expressed in MDCK cells were analyzed by confocal microscopy and Transwell ascorbate transport assays. These studies identified an SVCT2 basolateral targeting sequence (BTS) in the N-terminus, which is conserved among mammalian SVCT2 forms. The less conserved N-terminus of SVCT1 is not required for apical localization. The destruction of SVCT2 BTS led to apical localization of the protein in a manner independent of the C-terminal sequence. A C-terminal sequence present in both SVCTs appears to be required for plasma membrane incorporation and retention as its deletion led to an increased level of intracellular appearance of both apically and basolaterally targeted SVCTs in the absence or presence of BTS. Nevertheless, all C-terminal deletion mutants showed preferential apical transport activity, suggesting a greater importance of this sequence for basolateral targeting. Our results collectively suggested a default apical targeting of SVCT, which is consistent with the evolution-based prediction. The SVCT sorting model with a hierarchal contribution of N- and C terminal sequences was compared to the observations made for other multipass membrane proteins. The involvement of both intracellularly localized termini of multipass membrane proteins in the sorting pathway suggests a more complex sorting mechanism compared to that for single-pass proteins. PMID- 19216497 TI - Synthesis of 1-Boc-3-fluoroazetidine-3-carboxylic acid. AB - Synthetic strategies toward 3-fluoroazetidine-3-carboxylic acid, a new cyclic fluorinated beta-amino acid with high potential as building block in medicinal chemistry, were evaluated. The successful pathway includes the bromofluorination of N-(diphenylmethylidene)-2-(4-methoxyphenoxymethyl)-2-propenylamine, yielding 1 diphenylmethyl-3-hydroxymethyl-3-fluoroazetidine after reduction of the imino bond, ring closure, and removal of the 4-methoxybenzyl group. Changing the N protecting group to a Boc-group allows further oxidation to 1-Boc-3 fluoroazetidine-3-carboxylic acid, a new fluorinated heterocyclic amino acid. PMID- 19216498 TI - Cataluminescence-based array imaging for high-throughput screening of heterogeneous catalysts. AB - High-throughput screening of catalysts could dramatically improve performance and reduce costs in the discovery and study of various catalysts. Here we report a cataluminescence-based array imaging as a high-throughput screening technique in the combinatorial discovery of active catalysts for CO oxidation. This strategy is based on the fact that the CO oxidation generates cataluminescence emission on the surface of nanomaterials, whose intensity is correlated to the activity of the catalyst. To demonstrate the feasibility of the cataluminescence-based array imaging for high-throughput screening of catalysts, different nanosized metal catalysts supported on TiO(2) nanoparticles were prepared. These catalysts include monometallic Au, Pt, and the bimetallic Au-Pt heteroaggregate catalysts, at total metal loadings of 0.5%, 1.0%, and 2.5%, and with atomic ratios of 1:1, 1:2, and 2:1 (Au/Pt). A 4 x 4 array was integrated by depositing these nanosized catalysts onto the ceramic chip, and the brightness of each spot in the image was recorded. The catalytic activities of those catalysts for the CO oxidation were evaluated parallelly by both the cataluminescence imaging and the gas chromatography method. The correlation coefficient is 0.914 for the two techniques, indicating that the cataluminescence imaging technique can be applied for the evaluation of the catalytic activities. Moreover, fast evaluation of multiple catalysts at a series of working temperature can be achieved by this cataluminescene-based array imaging. With the development of nanotechnology as well as the catalyst industry, the cataluminescence-based array imaging will address its importance in the high-throughput screening of catalysts. PMID- 19216499 TI - Synthesis and applications of a light-fluorous glycosyl donor. AB - A new method using a light-fluorous glycosyl donor and an orthogonal tagging strategy to synthesize oligosaccharides and glycoconjugates has been developed. The glycosyl donor orthogonally protected with a C8F17-silyl tag and benzoyl groups was reacted with excess amounts of glycosyl acceptor. Fluorous solid-phase extraction separated the glycosylated product and unreacted glycosyl acceptor. This new protocol has high reaction efficiency and easy separation, which was demonstrated in the synthesis of an unprotected trisaccharide and an O glycosylated serine in this paper. PMID- 19216500 TI - Dual stereoselectivity in the dialkylzinc reaction using (-)-beta-pinene derived amino alcohol chiral auxiliaries. AB - (+)-Nopinone, prepared from naturally occurring (-)-beta-pinene, was converted to the two regioisomeric amino alcohols 3-MAP and 2-MAP in very good yield and excellent isomeric purity. Amino alcohol 3-MAP was synthesized by converting (+) nopinone to the corresponding alpha-ketooxime. This was reduced to the primary amino alcohol and was converted to the morpholino group through a simple substitution reaction. 3-MAP was characterized by X-ray crystallography, which displayed the rigidity of the pinane framework. Amino alcohol 2-MAP was prepared from its trans isomer 2, which in turn was synthesized via hydroboration/oxidation of the morpholine enamine of (+)-nopinone. Two dimensional NMR was used to characterize amino alcohol 2-MAP, and NOE was used to confirm its relative stereochemistry. These amino alcohols were employed as chiral auxiliaries in the addition of diethylzinc to benzaldehyde to obtain near quantitative asymmetric induction in the products. The use of 3-MAP yielded (S) phenylpropanol in 99% ee, and its regioisomer 2-MAP gave the opposite enantiomer, (R)-phenylpropanol, also in 99% ee. Other aromatic, aliphatic, and alpha,beta unsaturated aldehydes were implemented in this method, affording secondary alcohols in high yield and enantiomeric excess. Amino alcohols 2-MAP and 3-MAP were also found to be useful in the dimethylzinc addition reaction, both catalyzing the addition to benzaldehyde with nearly quantitative ee. Regioisomeric amino alcohols 2-MAP and 3-MAP, even though they were prepared from one enantiomer of nopinone, provide antipodal enantiofacial selectivity in the dialkylzinc addition reaction. This circumvents the necessity to synthesize amino alcohols derived from (-)-nopinone, which in turn requires the unnatural (+)-beta pinene. Possible mechanistic insights are offered to explain the dual stereoselectivity observed in the diethylzinc addition reaction involving regioisomeric, pseudo-enantiomeric amino alcohols 3-MAP and 2-MAP. PMID- 19216503 TI - Cyclization and ring-expansion processes involving samarium diiodide promoted reductive formation and subsequent oxidative ring opening of cyclopropanol derivatives. AB - Samarium diiodide promoted reaction of various alpha-bromomethyl cycloalkanones, followed by subsequent treatment with trimethylsilyl chloride, leads to the production of cyclopropyl silyl ethers embedded in bicyclo[m.1.0]alkane frameworks. Treatment of the ethers with oxidative electron-transfer reagents, such as Fe(III), Ce(IV), and Mn(III) salts, generates ring-expanded ketones that convert to cyclic conjugated enones in moderate to good yields. In addition, the reduction-oxidation reaction sequences can be successfully performed in one pot. The regioselectivities of cyclopropane ring opening in the bicyclic substrates depend on the oxidizing agents used. For example, reactions promoted by FeCl3 with pyridine lead to the expected ring-expansion process involving internal-bond cleavage of bicycloalkane and yielding cyclic enones as final products. In contrast, reactions with Ce(NH4)2(NO3)6 or Mn(OAc)3 as oxidizing agents proceed by way of external-bond cleavage to give alpha-iodomethyl cycloalkanones. PMID- 19216504 TI - On the mechanism of the mutagenic action of 5-bromouracil: a DFT study of uracil and 5-bromouracil in a water cluster. AB - Density functional theory calculations on the canonical (keto) and rare (enol) tautomeric forms of uracil and 5-bromouracil in a cluster consisting of 50 water molecules are presented. The keto form of uracil is favored over the enol tautomer in both the gas phase and solution. However, the presence of the water cluster reverses the tautomeric preference of 5-bromouracil, rendering the rare tautomeric form to be preferred over the canonical form in aqueous solution. This effect is, to a large extent, due to the more favorable water-water interactions in the cluster around 5-bromouracil and can therefore only be obtained by including explicit water-water interactions in the calculations. PMID- 19216505 TI - Reaching for the other side: generating sequence-dependent interstrand cross links with 5-bromodeoxyuridine and gamma-rays. AB - Interstrand cross-links impede critical cellular processes such as transcription and replication and are thus considered to be one of the most toxic types of DNA damage. Although several studies now point to the existence of gamma-radiation induced cross-links in cellular DNA, little is known about the characteristics required for their creation. Recently, we reported the formation of interstrand cross-links that were specific for mismatched nucleotides within 5-bromo-2' deoxyuridine-substituted DNA. Given the structural specificity for interstrand cross-link formation, it is likely that open or mismatched regions of DNA in cells may be particularly favorable for cross-link production. Herein, we investigated the effect of the local DNA sequence on the formation of interstrand cross-links, using 5-bromo-2'-deoxyuridine to generate radicals in a mismatched region of DNA. We investigated a total of 12 variations of bases in the mismatched region. The oligonucleotides were irradiated with gamma-rays, and interstrand cross-link formation was analyzed by denaturing gel electrophoresis. We found that the efficiency of cross-link formation was highly dependent on the nature of mismatched bases and, on the basis of electrophoretic mobility, observed several distinctive cross-link structures with specific DNA sequences. This study provides new insights into the reactivity of mismatched DNA and the mechanisms leading to interstrand cross-link formation. The potential application of 5-bromo-2'-deoxyuridine-induced interstrand cross-links to the field of DNA repair is discussed. PMID- 19216506 TI - Addition of H2O and O2 to acetone and dimethylsulfoxide ligated uranyl(V) dioxocations. AB - Gas-phase complexes of the formula [UO2(lig)]+ (lig = acetone (aco) or dimethylsulfoxide (dmso)) were generated by electrospray ionization (ESI) and studied by tandem ion-trap mass spectrometry to determine the general effect of ligand charge donation on the reactivity of UO2(+) with respect to water and dioxygen. The original hypothesis that addition of O2 is enhanced by strong sigma donor ligands bound to UO2(+) is supported by results from competitive collision induced dissociation (CID) experiments, which show near exclusive loss of H2O from [UO2(dmso)(H2O)(O2)]+, whereas both H2O and O2 are eliminated from the corresponding [UO2(aco)(H2O)(O2)]+ species. Ligand-addition reaction rates were investigated by monitoring precursor and product ion intensities as a function of ion storage time in the ion-trap mass spectrometer: these experiments suggest that the association of dioxygen to the UO2(+) complex is enhanced when the more basic dmso ligand was coordinated to the metal complex. Conversely, addition of H2O is favored for the analogous complex ion that contains an aco ligand. Experimental rate measurements are supported by density function theory calculations of relative energies, which show stronger bonds between UO2(+) and O2 when dmso is the coordinating ligand, whereas bonds to H2O are stronger for the aco complex. PMID- 19216507 TI - Axial asymmetry of the charge- and spin-density distributions in Pi states. Molecular quadrupole moments and hyperfine coupling constants of CH, NH, OH, CF, LiO, NO, and FO. AB - The axial asymmetry of the charge- and spin-density distributions in Pi states is studied via second-rank traceless tensors P(ii) (ii = xx, yy, zz), namely, quadrupole moments (Theta(ii)), electric field gradients (q(ii)), and magnetic dipolar (T(ii)) hyperfine coupling constants (hfcc's). In linear molecules, it holds that P(xx) does not = P(yy) does not = P(zz) for Pi, but P(xx) = P(yy) does not = P(zz) for Sigma, Delta, Phi,..., states. Thus, traceless P(ii) in Pi states have two independent parameters, P(parallel) = P(zz) is proportional to [r(m)(3 cos2 theta - 1] and deltaP(perpendicular) = |P(xx) - P(yy)| is proportional to [r(m) sin2 theta], with m = 2(Theta(ii)) or -3(q(ii), T(ii)). All linear states have P(parallel) does not = 0, but only Pi states exhibit deltaP(perpendicular) does not = 0, as shown by hfcc's like c = (3/2)T(zz), and d = |T(xx) - T(yy)|, as well as q0 = (-q(zz)) and |q(2)/2| = |q(xx) - q(yy)|. Little is known about Theta(zz) and deltaTheta(perpendicular) = |Theta(xx) - Theta(yy)| in Pi states since most experimental values (gas-phase) are rotational averages, and several theoretical studies have reported Theta(zz) but assumed deltaTheta(perpendicular) = 0. The diatomics studied here have X2Pi(1/2)(pi1) ground states, like CH and NO, or are of type X2Pi(3/2)(pi3), like OH, CF, LiO, and FO. The A3Pi(sigma pi3) state of NH is also included. Our P(parallel) and deltaP(perpendicular) values- calculated at the experimental R(e)'s with the B3LYP/aug-cc-pVQZ method- reproduce well the available literature data. The properties of the CF and FO radicals are not well-known so that our {c, d} and {q0, q2} values should help future experimental studies of their hyperfine spectra. Excluding OH, the complete quadrupole sets {Theta(zz), deltaTheta(perpendicular)} are new for all species discussed here. For comparison purposes, Theta(zz) of a low-lying Sigma state is also calculated for each X2Pi radical. PMID- 19216509 TI - Effect of sulfur oxidation on the transmission mechanism of 4J(HH) NMR coupling constants in 1,3-dithiane. AB - Long-range 4J(HH) couplings in 1,3-dithiane derivatives are rationalized in terms of the effects of hyperconjugative interactions involving the S=O group. Theoretical and experimental studies of 4J(HH) couplings were carried out in 1,3 dithiane-1-oxide (2), cis-1,3-dithiane-1,3-dioxide (3), 1,3-dithiane-1,1,3 trioxide (4), and 1,3-dithiane-1,1,3,3-tetraoxide (5) compounds. Hyperconjugative interactions were studied with the natural bond orbital, NBO, method. Hyperconjugative interactions involving the LP(O), oxygen lone pair and sigma*(C2 S1) and sigma*(S1-C6) antibonding orbitals yield an increase of 4J(H(eq)-H(eq)) couplings. Long-range 4J(H(ax)-H(ax)) couplings were also observed between hydrogen atoms in axial orientation, which are rationalized as originating in hyperconjugative interactions involving the bonding sigma(C6-H(ax)) and antibonding sigma*(S=O) orbitals. The symmetry for orbital interactions is possible only when the S=O group is in the axial orientation. PMID- 19216510 TI - Formation of monomeric S100B and S100A11 proteins at low ionic strength. AB - The S100 proteins comprise a group of EF-hand proteins that undergo a calcium induced conformational change allowing them to interact with other proteins and produce a biological response. A unique feature of these proteins is the fact that they can form both homo- and heterodimers independent of calcium binding. The reported dissociation constants for several S100 proteins span a very large range, from 1-4 microM to <<1 nM, suggesting that differing interface surface areas could govern the strength of the binding affinity. In this work, we examine the dimerization mechanism of S100B and S100A11 in the absence of calcium. Using electrospray mass spectrometry, we demonstrate that the monomer-dimer equilibrium in these S100 proteins is strongly dependent on the ionic strength of the solution. At higher ionic strengths (>or=22 mM), both S100A11 and S100B exist predominantly as homodimers. For apo-S100A11, a K(dimer) near 0.01 microM is estimated, while concentration-dependent experiments under these conditions show the K(dimer) for apo-S100B must be even lower. In contrast, lowering the ionic strength results in the formation of monomeric proteins with poorer dimer propensity. For example, the estimated K(dimer) for apo-S100A11 is more than 400 microM at 0.1 mM NH(4)Ac. (1)H-(15)N HSQC NMR experiments in combination with circular dichroism studies show that monomeric S100B and S100A11 proteins are alpha-helical and retain a significant amount of tertiary structure. Our results indicate that apo-S100B has at least a 10-fold stronger propensity to form dimers than does apo-S100A11 in line with a 400 A(2) greater buried surface area for apo S100B at its dimer interface. These experiments are the first to show that folded monomeric S100 proteins can be isolated, thus paving the way for future experiments aimed at examining the possible role of these monomers in folding and calcium signaling. PMID- 19216511 TI - Structural, spectroscopic, and thermodynamic consequences of anti-chelate effect in nine-coordinate lanthanide podates. AB - The connection of three tridentate 2,6-bis(1-ethyl-benzimidazol-2-yl)pyridine binding units to an extended sulfur-containing tripodal anchor in the ligand L9 yields nine-coordinate podates [Ln(L9)](3+) upon reaction with trivalent lanthanides, Ln(III). Structural analysis of [Eu(L9)](ClO(4))(3) in the solid state with the help of the bond valence method shows that the peripheral ethyl groups are responsible for a specific distortion of the triple-helical structure, which allows a closer approach of the nitrogen donors toward the central metal, while minimizing interstrand repulsion. The consequences of this distortion on the Eu(III) luminescent probe are investigated by high-resolution emission spectroscopy, while paramagnetic NMR data collected in acetonitrile demonstrate that [Ln(L9)](3+) adopts a single relaxed C(3)-symmetrical structure along the complete lanthanide series. The persistence of the triple-helical structure in solution is obtained at the cost of severe constraints in the helically wrapped organic tripod, which strongly disfavor intramolecular cyclization processes. The resulting antichelate effect can be exploited for the selective preparation of polynuclear complexes with tripodal ligands. PMID- 19216512 TI - Effect of imidazole and phenolate axial ligands on the electronic structure and reactivity of oxoiron(IV) porphyrin pi-cation radical complexes: drastic increase in oxo-transfer and hydrogen abstraction reactivities. AB - To study the effect of axial ligands on the electronic structure and reactivity of compound I of peroxidases and catalases, oxoiron(IV) porphyrin pi-cation radical complexes with imidazole, 2-methylimidazole, 4(5)-methylimidazole, and 3 fluoro-4-nitrophenolate as the axial ligands were prepared by ozone oxidation of iron(III) complexes of 5,10,15,20-tetramesitylporphyrin (TMP) and 2,7,12,17 tetramethyl-3,8,13,18-tetramesitylporphyrin (TMTMP). These complexes were fully characterized by absorption, (1)H, (2)H, and (19)F NMR, electron paramagnetic resonance (EPR), and electrospray ionization mass spectrometry (ESI-MS) spectroscopy. The characteristic absorption peak of compound I at approximately 650 nm was found to be a good marker for estimation of the electron donor effect from the axial ligand. The axial ligand effect did not change the porphyrin pi cation radical state, the a(2u) state of the TMP complexes, or the a(1u) radical state of both the TMTMP complexes and compound I. The ferryl iron and porphyrin pi-cation radical spins were effectively transferred into the axial ligands for the a(2u) complexes but not for the a(1u) complexes. Most importantly, the reactivity of the oxoiron(IV) porphyrin pi-cation radical complex was drastically increased by the imidazole and phenolate axial ligands. The reaction rate for cyclooctene epoxidation was increased 100- to 400-fold with axial coordination of imidazoles and phenolate. A similar increase was also observed for the oxidation of 1,4-cyclohexadiene,N,N-dimethyl-p-nitroaniline and hydrogen peroxide. These results suggest extreme enhancement of the reactivity of compound I by the axial ligand in heme enzymes. The functional role of axial ligands on the compound I in heme enzymes is discussed. PMID- 19216513 TI - Redox isomerism in the lanthanide complex [(dpp-Bian)Yb(DME)(mu-Br)]2 (dpp-Bian = 1,2-bis[(2,6-diisopropylphenyl)imino]acenaphthene). AB - Ytterbium reacts with 1,2-bis[(2,6-diisopropylphenyl)imino]acenaphthene (1, dpp Bian) in 1,2-dimethoxyethane (DME) to give complex (dpp-Bian)Yb(DME)(2) (2). Oxidation of 2 with an 0.5 mol equivalent of dibromostilbene affords dimeric compound [(dpp-Bian)Yb(DME)(mu-Br)](2) (3). Molecular structures of 2 and 3 were determined by single-crystal X-ray analysis. In complex 3 in a DME solution, a temperature-dependent reversible intramolecular electron transfer between the ligand and the metal takes place. PMID- 19216514 TI - Stereoselective one-pot, three-component synthesis of 4-aryltetrahydropyran via Prins-Friedel-Crafts reaction. AB - A diastereoselective one-pot, three-component Prins-Friedel-Crafts reaction was developed for the synthesis of 4-aryltetrahydropyran derivatives from the reaction of carbonyl compounds with homoallylic alcohol in the presence of arene promoted by boron trifluoride etherate. PMID- 19216515 TI - Calculation and meaning of feasible band boundaries in multivariate curve resolution of a two-component system. AB - Results obtained by model-free multivariate curve resolution (MCR) methods often are complicated by rotational and scale ambiguities, meaning that a range of feasible solutions describing and fitting experimental data equally well and fulfilling the constraints of the system are possible. In this work, two recent proposals to examine this problem and their relation are compared and discussed for the case of a two-component system. In one of these approaches, a systematic grid search of all feasible solutions is performed, and the results are displayed in appropriate mesh and contour plots which reveal their boundaries. In a second approach, an objective function is defined in terms of the relative signal contribution of every chemical species, and this function is maximized and minimized to get its extreme values that satisfy the constraints. These extreme values can also be represented graphically in the previously obtained mesh and contour plots. It turns out that the results obtained by these two approaches are in agreement and that the same extreme values are identified as boundaries of the band of feasible solutions, proving their reliability and their possible general application for the validation of MCR results. PMID- 19216516 TI - Structural characterization of a soluble amyloid beta-peptide oligomer. AB - Alzheimer's disease (AD) is a neurodegenerative disorder that is linked to the presence of amyloid beta-peptides that can form insoluble fibrils or soluble oligomeric assemblies. Soluble forms are present in the brains and tissues of Alzheimer's patients, and their presence correlates with disease progression. Long-lived soluble forms can be generated in vitro by using small amounts of aliphatic hydrocarbon chains of detergents or fatty acids in preparations of amyloid beta-peptides. Using NMR, we have characterized soluble oligomers of Abeta preglobulomer and globulomer that are stable and alter synaptic activity. The NMR data indicate that these soluble forms have a mixed parallel and antiparallel beta-sheet structure that is different from fibrils which contain only parallel beta-sheets. Using the structural data, we engineered a disulfide bond into the soluble Abeta globulomer to give a "new" soluble antigen that is stable, homogeneous, and binds with the same affinity to selective antibodies as the parent wt globulomer. PMID- 19216517 TI - Antineoplastic agents. 571. Total synthesis of bacillistatin 2. AB - The first total synthesis of bacillistain 2 (2) has been achieved in 24 steps and 22.9% overall yield, providing a quite efficient route with maximal convergence. Notable features of this approach include two successful applications of the Mitsunobu reaction during respective assemblies of key intermediates 22 and 27, successful employment of 2-methyl-6-nitrobenzoic anhydride (MNBA) in the formation by lactonization of a macrocyclic (36-membered) ring, and very flexible access to structural modifications of the bacillistatin-type cyclodepsipeptides. PMID- 19216518 TI - High-affinity labeling and tracking of individual histidine-tagged proteins in live cells using Ni2+ tris-nitrilotriacetic acid quantum dot conjugates. AB - Investigation of many cellular processes using fluorescent quantum dots (QDs) is hindered by the nontrivial requirements for QD surface functionalization and targeting. To address these challenges, we designed, characterized and applied QD trisNTA, which integrates tris-nitrilotriacetic acid, a small and high-affinity recognition unit for the ubiquitous polyhistidine protein tag. Using QD-trisNTA, we demonstrate two-color QD tracking of the type-1 interferon receptor subunits in live cells, potentially enabling direct visualization of protein-protein interactions at the single molecule level. PMID- 19216519 TI - Biosynthesis of the thiamin thiazole in Bacillus subtilis: identification of the product of the thiazole synthase-catalyzed reaction. AB - In this paper, we describe an optimized reconstitution of the thiamin thiazole synthase (ThiG) catalyzed reaction and demonstrate that the enzymatic product is an unanticipated dearomatized thiazole tautomer. PMID- 19216520 TI - Saldedines A and B, dibromo proaporphine alkaloids from a Madagascan tunicate. AB - Two new dibromo proaporphine alkaloids, designated saldedines A (1) and B (2), were isolated from an unidentified tunicate collected at Salary Bay, Madagascar. Saldedines A and B are the first marine proaporphine alkaloids. Their structures were elucidated by extensive spectroscopic means, and the structure of 1 was confirmed by single-crystal X-ray diffraction analysis. Both saldedines A and B were tested for toxicity to brine shrimp and showed moderate activity. PMID- 19216521 TI - Pd(0)/InI-mediated allylic additions to 4-acetoxy-2-azetidinone: new route to highly functionalized carbocyclic scaffolds. AB - Acylnitroso-derived hetero-Diels-Alder cycloadducts are susceptible to C-O bond cleavage with Pd(0) and InI to form allylic indium(III) species. The in situ prepared allylindium compounds readily react at room temperature with Eschenmoser's salt. Allylation of 4-acetoxy-2-azetidinone provides derivatized cyclopentenes in high regio- and diastereoselectivity. PMID- 19216522 TI - Solvent-induced high fidelity switching between two discrete supramolecules. AB - Here we show the reversible high fidelity switching between two discrete self assembled supramolecules made from a lipophilic 8-phenyl-2'-deoxyguanosine derivative induced by an indirect solvent effect. A hexadecameric supramolecule containing four stacked tetramers is formed in acetonitrile aided by higher potassium concentrations. When the amount of weakly solvated potassium decreases, due the lower activity of potassium iodide in chloroform, an octamer is formed after the dissociation of the two outer tetramers in the hexadecamer. The switching process results from an unprecedented subtle interplay between the activity of potassium iodide and the steric crowding within the self-assembled structure. Besides the possible applications in nanoconstruction, this phenomenon sheds light into the mechanism of formation of self-assembled supramolecules made from guanosine derivatives. PMID- 19216523 TI - Controlled reversible immobilization of Ru carbene on single-walled carbon nanotubes: a new strategy for green catalytic systems based on a solvent effect on pi-pi interaction. AB - A pyrene-tagged ruthenium carbene 8 was synthesized and immobilized on single walled carbon nanotubes (SWNTs) via pi-pi stacking. These pi-pi interactions were greatly affected by the reaction temperature and the solvent polarity, thus, offering a new reversible immobilization model that can be controlled by reaction temperature in polar solvents, such as acetone. SWNTs-supported ruthenium carbene 8 is a robust and recyclable catalyst system. Six to seven cycles were achieved for ring-closing metathesis of selected substrates. Importantly, after the complete loss of activity, the SWNTs can be easily recycled by washing with tetrahydrofuran. The recycled SWNTs can then be reloaded with 8. The reusability of the catalyst supported on recycled SWNTs is comparable with that supported on fresh SWNTs. PMID- 19216524 TI - An efficient synthesis of 2,5-dihydrofuran-fused bicyclic skeletons via the Pd(II)-catalyzed tandem-cyclization reaction of 1,omega-bisallenols. AB - A palladium(II)-catalyzed tandem double-cyclization reaction of 1,omega bisallenols was developed to form 2,5-dihydrofuran-fused bicyclic skeletons. With "unsymmetric" substrates, the reaction may also be realized with one hydroxyl group being protected as the acetate. Optically active bicyclic products were prepared by applying the Novozym-435 catalyzed kinetic resolution and the tandem double cyclization of these optically active allenol-allenyl acetates. The reaction may proceed via an oxypalladation, insertion, and elimination process. PMID- 19216526 TI - Self-assembled simple hexagonal AB(2) binary nanocrystal superlattices: SEM, GISAXS, and defects. AB - Binary superlattices (BSLs) of sterically stabilized, hydrophobic, large (A; 11.5 nm diameter) Fe(2)O(3) and small (B; 6.1 nm diameter) Au nanocrystals were assembled by slow evaporation of colloidal dispersions on tilted substrates. A detailed analysis of the BSL structure was carried out using transmission and scanning electron microscopy (TEM and SEM) and grazing incidence small-angle X ray scattering (GISAXS). The BSLs were simple hexagonal (sh) AB(2) superlattices (isostructural with the compound AlB(2); space group 191, P6/mmm) of large nanocrystals occupying a simple hexagonal lattice with small nanocrystals in the interstitial spaces. SEM and GISAXS confirmed long-range order of the BSLs and GISAXS revealed that the superlattice is slightly contracted (8-12%) perpendicular to the substrate as a result of solvent drying in the deposition process. When the sh-AB(2) superlattice deposits on a (100) plane, this shrinkage occurs in the [210] direction and changes the lattice symmetry to centered orthorhombic. Additionally, nearly periodic superlattice dislocations consisting of inserted half-planes of gold nanocrystals were observed by SEM in some BSLs. PMID- 19216527 TI - Toluene and ethylbenzene aliphatic C-H bond oxidations initiated by a dicopper(II)-mu-1,2-peroxo complex. AB - With an anisole-containing polypyridylamine potential tetradentate ligand (O)L, a mu-1,2-peroxo-dicopper(II) complex [{(O)LCu(II)}(2)(O(2)(2-))](2+) forms from the reaction of the mononuclear compound [Cu(I)((O)L)(MeCN)]B(C(6)F(5))(4) ((O)LCu(I)) with O(2) in noncoordinating solvents at -80 degrees C. Thermal decay of this peroxo complex in the presence of toluene or ethylbenzene leads to rarely seen C-H activation chemistry; benzaldehyde and acetophenone/1-phenylethanol mixtures, respectively, are formed. Experiments with (18)O(2) confirm that the oxygen source in the products is molecular O(2) and deuterium labeling experiments indicate k(H)/k(D) = 7.5 +/- 1 for the toluene oxygenation. The O(2) reaction of [Cu(I)((Bz)L)(CH(3)CN)](+) ((Bz)LCu(I)) leads to a dicopper(III)-bis mu-oxo species [{(Bz)LCu(III)}(2)(mu-O(2-))(2)](2+) at -80 degrees C, and from such solutions, very similar toluene oxygenation chemistry occurs. Ligand (Bz)L is a tridentate chelate, possessing the same moiety found in (O)L, but without the anisole O-atom donor. In these contexts, the nature of the oxidant species in or derived from [{(O)LCu(II)}(2)(O(2)(2-))](2+) is discussed and likely mechanisms of reaction initiated by toluene H-atom abstraction chemistry are detailed. To confirm the structural formulations of the dioxygen-adducts, UV-vis and resonance Raman spectroscopic studies have been carried out and these results are reported and compared to previously described systems including [{Cu(II)((Py)L)}(2)(O(2))](2+) ((Py)L = TMPA = tris(2-methylpyridyl)amine). Using (L)Cu(I), CO-binding properties (i.e., nu(C-O) values) along with electrochemical property comparisons, the relative donor abilities of (O)L, (Bz)L, and (Py)L are assessed. PMID- 19216528 TI - Quaternary ammonium beta-cyclodextrin nanoparticles for enhancing doxorubicin permeability across the in vitro blood-brain barrier. AB - This study describes novel quaternary ammonium beta-cyclodextrin (QAbetaCD) nanoparticles as drug delivery carriers for doxorubicin (DOX), a hydrophobic anticancer drug, across the blood-brain barrier (BBB). QAbetaCD nanoparticles show 65-88 nm hydrodynamic radii with controllable cationic properties by adjusting the incorporated amount of quaternary ammonium group in their structure. ATR-FTIR studies confirm the complexation between the QAbetaCD nanoparticles and DOX. QAbetaCD nanoparticles are not toxic to bovine brain microvessel endothelial cells (BBMVECs) at concentrations up to 500 microg x mL( 1). They also do not change the integrity of BBMVEC monolayers, an in vitro BBB model, including transendothelial electrical resistance value, Lucifer yellow permeability, tight junction protein occludin and ZO-1 expression and morphology, cholesterol extraction, and P-glycoprotein (P-gp) expression and efflux activity, at a concentration of 100 microg x mL(-1). Some QAbetaCD nanoparticles not only are twice as permeable as dextran (M(w) = 4000 g x mol(-1)) control, but also enhance DOX permeability across BBMVEC monolayers by 2.2 times. Confocal microscopy and flow cytometry measurements imply that the permeability of QAbetaCD nanoparticles across the in vitro BBB is probably due to endocytosis. DOX/QAbetaCD complexes kill U87 cells as effectively as DOX alone. However, QAbetaCD nanoparticles completely protect BBMVECs from cytotoxicity of DOX at 5 and 10 microM after 4 h incubation. The developed QAbetaCD nanoparticles have great potential in safely and effectively delivering DOX and other therapeutic agents across the BBB. PMID- 19216529 TI - Stability of artificial oil bodies constituted with recombinant caleosins. AB - Caleosin is a unique calcium binding protein anchoring to the surface of seed oil bodies by its central hydrophobic domain composed of an amphiphatic alpha-helix and a proline-knot subdomain. Stable artificial oil bodies were successfully constituted with recombinant caleosin overexpressed in Escherichia coli. The stability of artificial oil bodies was slightly or severely reduced when the amphiphatic alpha-helix or proline-knot subdomain in the hydrophobic domain of caleosin was truncated. Deletion of the entire central hydrophobic domain substantially increased the solubility of the recombinant caleosin, leading to a complete loss of its capability to stabilize these oil bodies. A recombinant protein engineered with the hydrophobic domain of caleosin replaced by that of oleosin, the abundant structural protein of seed oil bodies, could stabilize the artificial oil bodies, in terms of thermo- and structural stability, as effectively as caleosin or oleosin. PMID- 19216530 TI - Safety assessment and detection method of genetically modified Chinese Kale (Brassica oleracea cv. alboglabra ). AB - Sporamins are tuberous storage proteins and account for 80% of soluble protein in sweet potato tubers with trypsin-inhibitory activity. The expression of sporamin protein in transgenic Chinese kale (line BoA 3-1) conferred insecticidal activity toward corn earworm [ Helicoverpa armigera (Hubner)] in a previous report. In this study, we present a preliminary safety assessment of transgenic Chinese kale BoA 3-1. Bioinformatic and simulated gastric fluid (SGF) analyses were performed to evaluate the allergenicity of sporamin protein. The substantial equivalence between transgenic Chinese kale and its wild-type host has been demonstrated by the comparison of important constituents. A reliable real-time polymerase chain reaction (PCR) detection method was also developed to control sample quality. Despite the results of most evaluations in this study being negative, the safety of sporamin in transgenic Chinese kale BoA 3-1 was uncluded because of the allergenic risk revealed by bioinformatic analysis. PMID- 19216531 TI - RAPD-based method for the quality control of Mediterranean oregano and its contribution to pharmacognostic techniques. AB - A pharmacognostic survey of 84 commercial samples of Mediterranean oregano, obtained from wholesale traders between 2001 and 2007, pinpointed the presence of extraneous plant material in 90.5% of the samples. In 59% of them extraneous material of plant origin was above 20%. Two major groups of botanical foreign matter were identified: oregano-like flavored plants ( Satureja montana L., Origanum majorana L.) and plants lacking a clearly detectable essential oil profile ( Rubus sp., Cistus incanus L., Rhus coriaria L.), added as bulk extraneous material. A random amplified polymorphic DNA (RAPD) method was developed to make the detection of the second group of adulterants easier and speed pharmacognostic analysis of large batches of samples. Thirteen primers discriminating between Origanum spp. and Rubus caesius , R.coriaria, and C. incanus were individuated, allowing their detection in oregano samples with a limit of detection of 1%. The utilization of RAPD as a reliable test to probe the authenticity of Mediterranean oregano or previously screen the presence of specific contaminants is proposed as a complementary approach to pharmacognostic and phytochemical screening. PMID- 19216532 TI - Characterization of five typical agave plants used to produce mezcal through their simple lipid composition analysis by gas chromatography. AB - Five agave plants typically used in Mexico for making mezcal in places included in the Denomination of Origin (Mexican federal law that establishes the territory within which mezcal can be produced) of this spirit were analyzed: Agave salmiana ssp. crassispina, A. salmiana var. salmiana, Agave angustifolia, Agave cupreata, and Agave karwinskii. Fatty acid and total simple lipid profiles of the mature heads of each plant were determined by means of a modified Bligh-Dyer extraction and gas chromatography. Sixteen fatty acids were identified, from capric to lignoceric, ranging from 0.40 to 459 microg/g of agave. Identified lipids include free fatty acids, beta-sitosterol, and groups of mono-, di-, and triacylglycerols, their total concentration ranging from 459 to 992 microg/g of agave. Multivariate analyses performed on the fatty acid profiles showed a close similarity between A. cupreata and A. angustifolia. This fact can be ascribed to the taxa themselves or differences in growing conditions, an issue that is still to be explored. These results help to characterize the agaves chemically and can serve to relate the composition of mezcals from various states of Mexico with the corresponding raw material. PMID- 19216534 TI - The effect of temperature on the proteome of recombinant Pichia pastoris. AB - The impact of environmental factors on the productivity of yeast cells is poorly investigated so far. Therefore, it is a major concern to improve the understanding of cellular physiology of microbial protein production hosts, including the methylotrophic yeast Pichia pastoris. Two-Dimensional Fluorescence Difference Gel electrophoresis and protein identification via mass spectrometry were applied to analyze the impact of cultivation temperature on the physiology of a heterologous protein secreting P. pastoris strain. Furthermore, specific productivity was monitored and fluxes through the central carbon metabolism were calculated. Chemostat culture conditions were applied to assess the adaption to different growth temperatures (20, 25, 30 degrees C) at steady-state conditions. Many important cellular processes, including the central carbon metabolism, stress response and protein folding are affected by changing the growth temperature. A 3-fold increased specific productivity at lower cultivation temperature for an antibody Fab fragment was accompanied by a reduced flux through the TCA-cycle, reduced levels of proteins involved in oxidative stress response and lower cellular levels of molecular chaperones. These data indicate that folding stress is generally decreased at lower cultivation temperatures, enabling more efficient heterologous protein secretion in P. pastoris host cells. PMID- 19216533 TI - Lysine methylation of nuclear co-repressor receptor interacting protein 140. AB - Receptor interacting protein 140 (RIP140) undergoes extensive post-translational modifications (PTMs), including phosphorylation, acetylation, arginine methylation, and pyridoxylation. PTMs affect its subcellular distribution, protein-protein interaction, and biological activity in adipocyte differentiation. Arginine methylation on Arg(240), Arg(650), and Arg(948) suppresses the repressive activity of RIP140. Here, we find that endogenous RIP140 in differentiated 3T3-L1 cells is also modified by lysine methylation. Three lysine residues, Lys(591), Lys(653), and Lys(757), are mapped as potential methylation sites by mass spectrometry. Site-directed mutagenesis study shows that lysine methylation enhances its gene repressive activity. Mutation of lysine methylation sites enhances arginine methylation, while mutation on arginine methylation sites has little effect on its lysine methylation, suggesting a relationship between lysine methylation and arginine methylation. Kinetic analysis of PTMs of endogenous RIP140 in differentiated 3T3-L1 cells demonstrates sequential modifications on RIP140, initiated from constitutive lysine methylation, followed by increased arginine methylation later in differentiation. This study reveals a potential hierarchy of modifications, at least for lysine and arginine methylation, which bidirectionally regulate the functionality of a nonhistone protein. PMID- 19216535 TI - PLIPS, an automatically collected database of protein lists reported by proteomics studies. AB - The spectrum of problems covered by proteomics studies range from the discovery of compartment specific cell proteomes to clinical applications, including the identification of diagnostic markers and monitoring the effects of drug treatments. In most cases, the ultimate results of a proteomics study are lists of proteins found to be present (or differentially present) at cell physiological conditions under study. Normally, the results are published directly in the article in one or several tables. In many cases, this type of information remains disseminated in hundreds of proteomics publications. We have developed a Web mining tool which allows the collection of this information by searching through full text papers and automatically selecting tables, which report a list of protein identifiers. By searching through major proteomics journals, we have collected approximately 800 independent studies published recently, which reported about 1000 different protein lists. On the basis of this data, we developed a computational tool PLIPS (Protein Lists Identified in Proteomics Studies). PLIPS accepts as input a list of protein/gene identifiers. With the use of statistical analyses, PLIPS infers recently published proteomics studies, which report protein lists that significantly intersect with a query list. PLIPS is a freely available Web-based tool ( http://mips.helmholtz muenchen.de/proj/plips ). PMID- 19216542 TI - Modular stereocontrolled assembly of R2Zn, cyclic enones and N-tert butanesulfinyl imines. AB - The assembly of a wide range of dialkylzincs, cyclic enones, and chiral N-tert butylsulfinyl imines in the presence of the appropriate phosphoramidite ligands allowed the formation of beta-amino ketones with three consecutive stereogenic centers in a stereocontrolled manner. The Baeyer-Villiger oxidation of the resulting amino ketones led to the corresponding aminolactones with excellent regio- and stereoselectivities. PMID- 19216536 TI - Intracellular adaptation of Brucella abortus. AB - Macrophages were infected with virulent Brucella abortus strain 2308 or attenuated strain 19. Intracellular bacteria were recovered at different times after infection and their proteomes compared. The virulent strain initially reduced most biosynthesis and altered its respiration; adaptations reversed later in infection. The attenuated strain was unable to match the magnitude of the virulent strain's adjustments. The results provide insight into mechanisms utilized by Brucella to establish intracellular infections. PMID- 19216543 TI - Palladium-catalyzed cross-coupling reactions of potassium alkenyltrifluoroborates with organic halides in aqueous media. AB - Potassium vinyl and alkenyltrifluoroborates are cross-coupled with aryl and heteroaryl bromides using 1 mol % Pd loading of 4-hydroxyacetophenone oxime derived palladacycle or Pd(OAc)2 as precatalysts, K2CO3 as base, and TBAB as additive and water reflux under conventional or microwave heating to afford styrenes, stilbenoids, and alkenylbenzenes. These borates can be cross-coupled diastereoselectively with allyl and benzyl chlorides using KOH as base in acetone water (3:2) at 50 degrees C and 0.1 mol % Pd loading, giving the corresponding 1,4-dienes and allylarenes, respectively. These simple phosphine-free reaction conditions allow the palladium recycling from the aqueous phase during up to five runs by extractive separation of the products, which contain 58-105 ppm of Pd. PMID- 19216546 TI - Net-like assembly of Au nanoparticles as a highly active substrate for surface enhanced Raman and infrared spectroscopy. AB - Anodic aluminum oxide (AAO) templates were employed to filtrate and assemble Au nanoparticles by the pressure difference method. It was found that the colloidal Au nanoparticles can be uniformly arranged as nanonet assembly on the AAO surface. The net-assembled Au nanoparticles are clean and closely packed with nanochains. Taking fullerene C60/C70 as probe molecules, high-quality surface enhanced Raman scattering (SERS) spectra were observed. The net-assembled Au nanoparticles even synchronously support the observation of surface-enhanced infrared absorption (SEIRA) spectra of the fullerene C60/C70. These results indicate that the AAO template filtrated with net-assembled Au nanoparticles is a highly active substrate for surface-enhanced spectroscopy. PMID- 19216548 TI - Theoretical study of internal conversion decay rates associated with the three lowest singlet electronic states in pyrazine. AB - The general expressions we recently derived for calculating internal conversion decay rate constants between two adiabatic and between two diabatic displaced distorted-rotated harmonic potential energy surfaces (including all vibrational modes) are now applied to determine the decay rate constants of 1B(2u)(pi pi*)- >1B(3u)(n pi*), 1B(2u)(pi pi*)-->1A(g), and 1B(3u)(n pi*)-->1A(g) internal conversions in pyrazine molecule. The minimal models with respect to the number and the types of vibrational modes are investigated for these processes. An exact expression for the adiabatic vibrational frequencies of the coupling modes in terms of interstate coupling constants and the parameters determining the diabatic potential energy surfaces are also derived. PMID- 19216549 TI - Syntheses and antigestagenic activity of mifepristone derivatives. AB - A series of mifepristone derivatives with different "linker groups" in position 4' of the phenyl ring in the 11beta-position of the steroid scaffold (2-41) have been synthesized. Their antigestagenic activites were determined in a cell-based assay (alkali phosphatase assay in T47-D breast cancer cells) and compared with that of the parent compound mifepristone. SAR and QSAR studies reveal the influence of both lipophilicity and partial charge based van der Waals surface area descriptors on biological activity. Within the series of compounds described in this study, three mifepristone derivatives are identified with considerably high antigestagenic activity. These compounds are regarded as useful starting materials for the synthesis of either physiologically stable or cleavable progesterone receptor-binding conjugates for therapeutic or diagnostic purposes. PMID- 19216550 TI - Applications of ultra-performance liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry on analysis of 138 pesticides in fruit- and vegetable-based infant foods. AB - The applications of ultra-performance liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC QqTOF) in the determination of 138 pesticides in fruit- and vegetable-based infant foods were investigated. Pesticides were extracted from infant foods using a procedure known as the quick, easy, cheap, effective, rugged, and safe (QuEChERS) method. UPLC QqTOF MS full-scan with a relatively high sensitivity proved to be an ideal tool for screening of a large number of pesticides in a single analysis. UPLC QqTOF MS/MS provided product ion spectra that allowed for unequivocal confirmation of pesticides. Quantification was achieved using matrix-matched standard calibration curves with isotopically labeled standards or a chemical analogue as internal standards. The method performance parameters that included overall recovery, intermediate precision, and measurement uncertainty were evaluated according to a designed experiment, that is, the nested design. Generally, about 90% of the pesticides studied had recoveries between 81 and 110%, 90% had intermediate precision of 9000 compound library of synthetic DNA binding molecules for selective binding to the consensus sequence of the transcription factor LEF-1 followed by assessment of the candidate compounds in a series of assays that characterized functional activity (disruption of DNA-LEF-1 binding) at the intended target and site (inhibition of intracellular LEF-1-mediated gene transcription) resulting in a desired phenotypic cellular change (inhibit LEF-1 driven cell transformation) provided two lead compounds: lefmycin-1 and lefmycin 2. The sequence of screens defining the approach assures that activity in the final functional assay may be directly related to the inhibition of gene transcription and DNA binding properties of the identified molecules. Central to the implementation of this generalized approach to the discovery of DNA binding small molecule inhibitors of gene transcription was (1) the use of a technically nondemanding fluorescent intercalator displacement (FID) assay for initial assessment of the DNA binding affinity and selectivity of a library of compounds for any sequence of interest, and (2) the technology used to prepare a sufficiently large library of DNA binding compounds. PMID- 19216570 TI - Stereoselective synthesis and osteogenic activity of subglutinols A and B. AB - Since clinically approved immunosuppressive drugs (e.g., cyclosporin A, FK506) possess dose-dependent biphasic effects that cause undesirable side effects on bone structure, including osteopenia, osteoporosis, and increased incidence of bone fractures, considerable effort has been devoted to the identification of immunosuppressive drugs that promote bone formation in a dose-dependent manner. Herein, we report the stereoselective synthesis of subglutinols A and B and present initial biological data showing the significant potential of subglutinol A as an immunosuppressive drug with dose-dependent osteogenic activity. We also show that activating protein 1 (AP-1) family transcription factors could be one of the key regulators for the anabolic activity of subglutinol A. Such drugs with dose-dependent osteogenic activity might help reduce bone-associated side effects and be clinically useful for bone tissue transplantation. PMID- 19216571 TI - Ultrasensitive immunosensor for cancer biomarker proteins using gold nanoparticle film electrodes and multienzyme-particle amplification. AB - A densely packed gold nanoparticle platform combined with a multiple-enzyme labeled detection antibody-magnetic bead bioconjugate was used as the basis for an ultrasensitive electrochemical immunosensor to detect cancer biomarkers in serum. Sensitivity was greatly amplified by synthesizing magnetic bioconjugates particles containing 7500 horseradish peroxidase (HRP) labels along with detection antibodies (Ab2) attached to activated carboxyl groups on 1 microm diameter magnetic beads. These sensors had sensitivity of 31.5 microA mL ng(-1) and detection limit (DL) of 0.5 pg mL(-1) for prostate specific antigen (PSA) in 10 microL of undiluted serum. This represents an ultralow mass DL of 5 fg PSA, 8 fold better than a previously reported carbon nanotube (CNT) forest immunosensor featuring multiple labels on carbon nanotubes, and near or below the normal serum levels of most cancer biomarkers. Measurements of PSA in cell lysates and human serum of cancer patients gave excellent correlations with standard ELISA assays. These easily fabricated AuNP immunosensors show excellent promise for future fabrication of bioelectronic arrays. PMID- 19216572 TI - Synthesis of titania nanosheets with a high percentage of exposed (001) facets and related photocatalytic properties. AB - Anatase TiO(2) nanosheets with highly reactive (001) facets exposed have been successfully synthesized by a facile hydrothermal route, taking advantage of a specific stabilization effect of fluorine ion on (001) facets. The percentage of highly reactive (001) facets in such TiO(2) nanosheets is very high (up to 89%). In addition, the as-prepared TiO(2) nanosheets exhibit excellent activity in the photocatalytic degradation of organic contaminants. PMID- 19216573 TI - Synthesis of CuInSe(2) nanocrystals with trigonal pyramidal shape. AB - The synthesis of monodisperse chalcopyrite (tetragonal) CuInSe(2) nanocrystals is reported. The nanocrystals have trigonal pyramidal shape, and they exhibited a common crystallographic orientation when drop-cast onto carbon substrates. A crystallographic model for the nanocrystals was developed. The nanocrystals are bounded by one polar {112} surface facet and three nonpolar {114} surface facets. PMID- 19216576 TI - Introduction: Human metabolites in safety testing (MIST) issue. PMID- 19216574 TI - Modulation of catalytic function by differential plasticity of the active site: case study of Trypanosoma cruzi trans-sialidase and Trypanosoma rangeli sialidase. AB - trans-Sialidase is an essential enzyme for Trypanosoma cruzi, the causative agent of Chagas' disease, to escape from the host immune system and to invade the host cells. Therefore, T. cruzi trans-sialidase (TcTS) presents a potential and appealing therapeutic target for this lethal disease. The availability of a structurally very similar enzyme with strict hydrolase activity (Trypanosoma rangeli sialidase, TrSA) provides us a unique opportunity to understand the determinants of their structure and catalytic mechanism. In this study, we compare the catalytic cleft plasticity of free (apo) and ligand-bound (holo) forms of the two enzymes using molecular dynamics simulations. We focus on the mouth of the catalytic cleft that is defined by two residues: W312 and Y119 in TcTS and W312 and S119 in TrSA. Our results indicate that TcTS has a very flexible, widely open catalytic cleft, mostly due to W312 loop motion, in apo form. However, when the catalytic cleft is occupied by a ligand, the flexibility and solvent exposure of TcTS is significantly reduced. On the other hand, TrSA maintains a more open catalytic cleft compared to its crystal structures in both apo and holo forms (and compared to TcTS in holo forms). The reduced solvent exposure of TcTS catalytic cleft might be partially or fully responsible for TcTS to be a less efficient hydrolase than TrSA. PMID- 19216579 TI - Approaches to the assessment of stable and chemically reactive drug metabolites in early clinical trials. AB - The recommendations of the FDA's final Guidance document on Safety Testing of Drug Metabolites provide a framework for devising preclinical toxicology assessment paradigms, where necessary, for human metabolites of small molecule drug candidates. Importantly, these recommendations carry implications for the qualitative and quantitative analysis of circulating drug metabolites in early human trials, which typically are performed without the benefit of a radiolabeled tracer. In this perspective, an approach to these goals is outlined based on recent work at Merck Research Laboratories involving the use of ultraperformance liquid chromatography-mass spectrometry analysis, performed on a high-resolution time-of-flight mass spectrometer, of first-in-human study plasma samples. With the aid of a fractional mass filtering algorithm, drug metabolites are distinguished from endogeneous background materials and subsequently identified on the basis of their accurate masses, product ion mass spectra, and computer assisted structure elucidation software routines. Semiquantitative analysis then is based on calibration of the MS response to each analyte with reference to radioactivity data from in vitro metabolic profiles. In the case of chemically reactive drug metabolites, which are excluded from consideration in the Guidance, a proactive approach is advocated whereby potent (low dose) drug candidates with only a limited propensity to form electrophilic intermediates are advanced into development. Overall, a decision on the need to conduct separate evaluation of the safety profile of a human drug metabolite(s) should take into consideration all of the available information on the compound of interest and be based on a case-by-case approach employing sound scientific principles. PMID- 19216580 TI - Qualitative analysis of the role of metabolites in inhibitory drug-drug interactions: literature evaluation based on the metabolism and transport drug interaction database. AB - Guidance from the Food and Drug Administration on drug interaction studies does not include a specific section on contributions of metabolites to observed inhibitory drug-drug interactions, and the quantitative role of drug metabolites in inhibitory drug-drug interactions is not presently known. The current work was undertaken to evaluate what fraction of inhibitors of common drug-metabolizing enzymes [cytochrome P450 (P450) 1A2, 2E1, 2D6, 2C9, 2C19, 2C8, 2B6, and 3A4] have circulating metabolites that may contribute to observed in vivo interactions. A literature analysis was conducted using the Metabolism and Transport Drug Interaction Database to identify all precipitants (i.e., inhibitors) that cause more than a 20% increase in the area under the plasma concentration-time curve (AUC) of marker substrates. The database, PubMed, and product labels were then used to determine whether circulating metabolites were present after administration of these inhibitors. Of the total of 129 inhibitors identified, 106 were confirmed to have metabolites that circulate in plasma. An additional 14 inhibitors were identified that are extensively metabolized but whose metabolites either have not been identified or have not been investigated. Hence, only 7% of the inhibitors did not have circulating metabolites. Of the 21 potent inhibitors (>or=5-fold increase in AUC) currently known, 17 had circulating metabolites, and the remaining four were all extensively metabolized. On the basis of available in vitro data, 24 of all of the inhibitors were mechanism-based inactivators of P450 enzymes, while 105 were characterized as reversible inhibitors. In vitro evaluation of inhibition potential was conducted for only 32% of the circulating metabolites of the inhibitors. In conclusion, circulating metabolites are often present with inhibitors of P450 enzymes, suggesting a need for increased efforts to characterize the inhibitory potency of metabolites of candidate drugs and for newer models for in vitro to in vivo extrapolations. PMID- 19216581 TI - Biotransformation of benzo[a]pyrene in Ahr knockout mice is dependent on time and route of exposure. AB - Benzo[a]pyrene (BP) is an ubiquitous environmental pollutant with potent mutagenic and carcinogenic properties. The Ah receptor (Ahr) is important in the metabolic activation of BP and is therefore central to BP-induced carcinogenesis. Although Ahr(-/-) mice are refractory to BP-induced carcinogenesis, higher levels of BP-DNA and -protein adducts were formed in them than in wild-type mice. These results indicated the presence of an Ahr-independent and/or a slower biotransformation of BP in Ahr knockout mice. To address this issue further, we have now performed a time-course experiment, with mice receiving a single oral dose of BP (100 mg/kg). Wild-type mice have an effective clearance of BP metabolites, mainly through 3-hydroxybenzo[a]pyrene and 9-hydroxybenzo[a]pyrene in the feces with reduced levels of DNA and protein adducts in the examined tissues. On the other hand, the Ahr(-/-) mice appear to have a lower metabolic clearance of BP resulting in increased levels of DNA and protein adducts and of unmetabolized BP. In addition, we have performed an administration route experiment and found that skin-exposed Ahr(-/-) mice showed lower levels of protein adducts along with markedly reduced P450 1B1 expression, but only in the exposed area, as compared with the wild-type mice. In addition, the systemic uptake of BP is increased in the Ahr(-/-) mice as compared with the wild-type mice. Hence, the lack of a functional Ah receptor results in an Ahr-independent biotransformation of BP with a slower clearance of BP and higher levels of DNA and protein adducts, but the distribution and levels of BP and BP-protein adducts are clearly dependent on the route of exposure. PMID- 19216584 TI - Dentistry and health care reform in the USA. PMID- 19216582 TI - Protein adsorption of ultrafine metal oxide and its influence on cytotoxicity toward cultured cells. AB - Many investigations about the cellular response by metal oxide nanoparticles in vitro have been reported. However, the influence of the adsorption ability of metal oxide nanoparticles toward cells is unknown. The aim of this study is to understand the influence of adsorption by metal oxide nanoparticles on the cell viability in vitro. The adsorption abilities of six kinds of metal oxide nanoparticles, namely, NiO, ZnO, TiO2, CeO2, SiO2, and Fe2O3, to Dulbecco's modified Eagle's medium supplemented with a 10% fetal bovine serum (DMEM-FBS) component such as serum proteins and Ca2) were estimated. All of the metal oxide nanoparticles adsorbed proteins and Ca2+ in the DMEM-FBS; in particular, TiO2, CeO2, and ZnO showed strong adsorption abilities. Furthermore, the influence of the depletion of medium components by adsorption to metal oxide nanoparticles on cell viability and proliferation was examined. The particles were removed from the dispersion by centrifugation, and the supernatant was applied to the cells. Both the cell viability and the proliferation of human keratinocyte HaCaT cells and human lung carcinoma A549 cells were affected by the supernatant. In particular, cell proliferation was strongly inhibited by the supernatant of TiO2 and CeO2 dispersions. The supernatant showed depletion of serum proteins and Ca2+ by adsorption to metal oxide nanoparticles. When the adsorption effect was blocked by the pretreatment of particles with FBS, the inhibitory effect was lost. However, in NiO and ZnO, which showed ion release, a decrease of inhibitory effect by pretreatment was not shown. Furthermore, the association of the primary particle size and adsorption ability was examined in TiO2. The adsorption ability of TiO2 depended on the primary particle size. The TiO2 nanoparticles were size dependently absorbed with proteins and Ca2+, thereby inducing cytotoxicity. In conclusion, the adsorption ability of metal oxide nanoparticles is an important factor for the estimation of cytotoxicity in vitro for low-toxicity materials. PMID- 19216585 TI - Stability of maxillary expansion and tongue posture. AB - OBJECTIVE: To evaluate the adaptive changes and the stability in tongue posture following rapid maxillary expansion (RME) in patients without any signs or symptoms of respiratory disturbances. MATERIALS AND METHODS: Growing subjects with maxillary constrictions and bilateral buccal crossbites were included in the treatment group (n = 20). A control group (n = 20) comprised subjects with normal dentoskeletal features. RME appliances were used in the treatment group, with an average active expansion of 15 +/- 2 days. Cephalometric radiographs were traced and digitized to evaluate static tongue posture before RME and 6.75 +/- 0.48 months after RME. Follow-up radiographic evaluations of 17 expansion cases were also performed after an average of 29.25 +/- 1.85 months. Independent and paired t-tests were conducted to evaluate changes in tongue posture within and between groups. RESULTS: Results revealed significant reductions of tongue-to-palate (P < .05) as well as hyoid bone-to-mandibular plane (P < .01) distances following RME. The new tongue posture was found to be stable during the follow-up period. CONCLUSIONS: A higher tongue posture can be obtained with RME in children with no reported respiratory disturbances. PMID- 19216586 TI - Antimicrobial effects of zinc oxide in an orthodontic bonding agent. AB - OBJECTIVE: To test the null hypothesis that the addition of zinc oxide (ZnO) has no effect on the antimicrobial benefits and shear bond strength of a light-cured resin-modified glass ionomer. MATERIALS AND METHODS: ZnO was added to Fuji Ortho LC to create mixtures of 13% ZnO and 23.1% ZnO. Specimen discs of the modified bonding agent were incubated with Streptococcus mutans for 48 hours in a disc diffusion assay that was used to measure zones of bacterial inhibition. In addition, brackets were bonded to bovine deciduous incisors with the modified bonding agents, and shear bond strength was evaluated with a universal testing machine. RESULTS: The modified samples showed that antimicrobial activity increased as the concentration of ZnO increased. There were significant differences (P < .05) in antimicrobial activity. Post hoc tests showed that the antibacterial effects were 1.6 times greater with 23.1% ZnO than with 13% ZnO. There was no difference between Transbond and 0% ZnO (the negative control). After 1 month of daily rinsing, the antibacterial effects of 23.1% ZnO and 13% ZnO decreased 65% and 77%, respectively, but both maintained significant effects over the negative controls. There were no significant differences (P = .055) in shear bond strength between any of the mixture comparisons. CONCLUSIONS: The incorporation of ZnO into Fuji Ortho LC added antimicrobial properties to the original compound without significantly altering the shear bond strength. ZnO holds potential for preventing decalcification associated with orthodontic treatment. PMID- 19216587 TI - Mechanical tensile stress effects on the expression of bone sialoprotein in bovine cementoblasts. AB - OBJECTIVE: To develop a new cementoblast culture method and to detect bone sialoprotein (BSP) expression in response to high and low mechanical tensile stress in cementoblast in vitro. MATERIALS AND METHODS: Cementoblasts were collected from the roots of newborn bovine teeth and were identified with cementum-derived attachment protein (CAP) antibody 3G9. Cell proliferation was evaluated by MTT [3-(4,5-dimethylthazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay, and mineralization was confirmed by von Kossa staining. Mechanical tensile stress was applied in vitro to the cementoblast with the use of a uniaxial four point bending system with 2000 or 4000 microstrains, at a frequency of 0.5 Hz for 3, 6, 12, 24, or 36 hours. BSP mRNA level was quantified by real-time quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). RESULTS: A large amount of cementoblast was observed to be expressing CAP. Cementoblasts had a proliferation tendency similar to that of osteoblasts but different from that of periodontal ligament (PDL) cells. Cementoblasts had the ability to become mineralized between osteoblasts and PDL cells. The mechanical tensile stress significantly up-regulated BSP mRNA expression, which reached a peak at 24 hours in both 2000 and 4000 microstrain groups (P < .01) and was tenfold and sixfold higher than that of controls, respectively. BSP expression dropped toward baseline levels at 36 hours in both groups. CONCLUSIONS: Mechanical tensile stress up-regulated the expression of BSP. Low mechanical tensile stress induced earlier and more intensive up-regulation of BSP mRNA; this might represent the optimal stimuli for cementoblast activity. PMID- 19216588 TI - Growth-related differences in maximum laterotrusion and retrusion between children and adults. AB - OBJECTIVE: To test the null hypothesis that there are no differences between children and adults in maximum laterotrusion and maximum retrusion on the right and left sides. MATERIALS AND METHODS: This population-based study included 81 randomly selected children between the ages of 6 and 10 years and 67 adults. Kinematic variables were measured with the ultrasonic JMA-System for registration. RESULTS: The mean maximum laterotrusion of the children's group (10.6 +/- 1.5 mm on the left, 11.0 +/- 1.7 mm on the right) was significantly smaller than that of the adult group (11.7 +/- 2.0 mm on the left, 12.2 +/- 1.7 mm on the right). The maximum laterotrusion of the children's group corresponded to about 90% on the left and right sides of that of the adult group. The mean maximum retrusion of the children's group was significantly bigger than that of the adult group. There, the adult values corresponded to 66.7% on the left and 50% on the right side of the children's values. No significant difference in maximum laterotrusion and retrusion was noted on the right and left sides, and no significant differences according to gender specificities were observed in either group. CONCLUSIONS: The hypothesis is rejected. In development of the temporomandibular joint, maximum laterotrusion on the right and left sides increases significantly with age, and maximum retrusion decreases significantly with age. PMID- 19216589 TI - Caries risk profiles in orthodontic patients at follow-up using Cariogram. AB - OBJECTIVE: To analyze caries-related factors shortly after orthodontic treatment and to use the Cariogram computer program to describe caries risk profiles at follow-up in these patients. MATERIALS AND METHODS: One hundred orthodontic patients age 12-29 years, with a mean age of 17.5 years, were included in the study. They were divided into two groups (50 in each) based on their prebonding decayed, filled surfaces index (DFS). High (5 > or = DFS) and low (2 < or = DFS) groups were created. All patients were examined after debonding in the following order: interview, plaque score, caries examination, saliva samples, bitewing radiographs, panoramic radiographs, and intra-oral digital photos. All types of carious lesions in both the enamel and dentine were diagnosed clinically and radiographically and included in the DFS index. A paraffin-stimulated whole saliva sample was collected for estimations of secretion rate, buffer capacity, and number of mutans streptococci and lactobacilli. RESULTS: The low caries group (2 < or = DFS) displayed a statistically significant difference and low values for the following factors, DFS (P < .001), lactobacilli (P < .001), mutans streptococci (P < .001), and high Cariogram percent (P < .001). The plaque index displayed very close significance (P = .051). CONCLUSIONS: Patients with high (5 > or = DFS) numbers before orthodontic treatment ran a higher risk of developing caries. They had significantly higher numbers of mutans streptococci and lactobacilli and had less chance of avoiding new cavities according to the Cariogram. PMID- 19216590 TI - Dentoskeletal and soft tissue effects of mini-implants in Class II division 1 patients. AB - OBJECTIVE: To examine the skeletal, dental, and soft tissue treatment effects of retraction of maxillary anterior teeth with mini-implant anchorage in nongrowing Class II division 1 female patients. MATERIALS AND METHODS: Twenty-three patients (overjet > or =7 mm) were selected on the basis of predefined selection criteria. Treatment mechanics consisted of retraction of anterior teeth by placing mini implants in the interdental bone between the roots of the maxillary first molar and second premolar. A force of 150 g was applied, bilaterally. Treatment effects were analyzed by taking lateral cephalograms and study casts at T1 (before initiation of retraction) and at T2 (after complete space closure). RESULTS: The upper anterior teeth showed significant retraction (5.18 +/- 2.74 mm) and intrusion (1.32 +/- 1.08 mm). The upper first molar also showed some distal movement and intrusion, but this was not significant (P > .05). The upper and lower lips were retracted by 2.41 mm and 2.73 mm, respectively, and the convexity angle reduced by over 2 degrees (P < .001). CONCLUSION: Mini-implants provided absolute anchorage to bring about significant dental and soft tissue changes in moderate to severe Class II division 1 patients and can be considered as possible alternatives to orthognathic surgery in select cases. PMID- 19216591 TI - Effects of Corticision on paradental remodeling in orthodontic tooth movement. AB - OBJECTIVE: To investigate the biologic effects of Corticision on alveolar remodeling in orthodontic tooth movement. MATERIALS AND METHODS: In this study, 16 cats were divided into 3 groups: group A, only orthodontic force (control); group B, orthodontic force plus Corticision; and group C, orthodontic force plus Corticision and periodic mobilization. Histologic and histomorphometric studies were performed on tissue specimens on days 7, 14, 21, and 28. RESULTS: Extensive direct resorption of bundle bone with less hyalinization and more rapid removal of hyalinized tissue were observed in group B. The accumulated mean apposition area of new bone on day 28 was observed to be 3.5-fold higher in group B than in the control group A. CONCLUSIONS: Corticision might be an efficient procedure for accelerating orthodontic tooth movement accompanied with alveolar bone remodeling. PMID- 19216592 TI - Surface characteristics of orthodontic materials and their effects on adhesion of mutans streptococci. AB - OBJECTIVE: To test the hypothesis that there are no significant differences in the adhesion of mutans streptococci (MS) to various orthodontic materials based on their surface characteristics. MATERIALS AND METHODS: Surface roughness (SR) and surface free energy (SFE) characteristics were investigated for nine different orthodontic materials (four orthodontic adhesives, three bracket raw materials, hydroxyapatite blocks, and bovine incisors) using confocal laser scanning microscopy and sessile drop method. Each material, except the bovine incisors, was incubated with whole saliva or phosphate-buffered saline for 2 hours. Adhesion assays were performed by incubating tritium-labeled MS with each material for 3 or 6 hours. RESULTS: Orthodontic adhesives had higher SFE characteristics and lower SR than bracket materials. Orthodontic adhesives showed a higher MS retaining capacity than bracket materials, and MS adhesion to resin modified glass ionomer and hydroxyapatite was highest. Extended incubation time increased MS adhesion, while saliva coating did not significantly influence MS adhesion. SFE, specifically its dispersive and polar components, was positively correlated with MS adhesion, irrespective of saliva coating. CONCLUSIONS: The hypothesis is rejected. This study suggests that SFE characteristics play an important role in the initial MS adhesion to orthodontic materials. PMID- 19216593 TI - Masticatory function and maturation of the jaw-opening reflex. AB - OBJECTIVE: To test the null hypothesis that alteration in masticatory function due to liquid-diet feeding during growth does not affect jaw-opening reflex (JOR) maturation. MATERIALS AND METHODS: Soon after weaning, 70 female Wistar rats were divided into two equal groups and fed either solid (control group) or liquid (experimental group) diets. At 5, 9, and 13 weeks, the rats were anesthetized and the JOR was recorded in the anterior belly of the digastric muscles as evoked by a low-intensity electrical stimulation of the left inferior alveolar nerve. RESULTS: There were similar tendencies at each recording age. Peak-to-peak amplitude of the JOR was significantly smaller, and the latency was significantly longer in the experimental group, although the duration was not significantly different between the two groups. CONCLUSIONS: These data suggest that long-term masticatory functional change due to liquid-diet feeding during growth may impede the learning for JOR maturation, and thus may affect the masticatory performance in the adult. PMID- 19216594 TI - Metal and ceramic bracket effects on human buccal mucosa epithelial cells. AB - OBJECTIVE: To test the null hypothesis that metal and ceramic brackets have no effect on the epithelial cells of the buccal mucosa. MATERIALS AND METHODS: Two metal and two ceramic brackets were bonded in 21 individuals of both sexes. With the use of liquid-based exfoliative cytology, morphometric and morphologic changes in buccal mucosa cells adjacent to these brackets were determined and were compared at three time points: baseline (T0), 60 days after placement (T1), and 30 days after removal of the brackets (T2). RESULTS: A decrease in nuclear area and an increase in cytoplasmic area occurred in the buccal mucosa cells adjacent to the brackets at T1 (P < .01). At T2, this altered morphometry persisted only in cells adjacent to the metal brackets, although to a lesser degree than at T1 (P < .01). A greater decrease in nuclear area was noted in cells adjacent to the metal brackets than in those next to the ceramic brackets (P < .01). At T0, the proportions of surface and subsurface cells were similar, but at T1, a predominance of surface cells was observed (P < .05). At all time points, smears of cells appeared normal or normal with some inflammatory changes. CONCLUSION: The hypothesis is rejected. Placement of metal and ceramic brackets in the buccal cavity induces cellular alterations. These alterations do not suggest malignancy. PMID- 19216595 TI - Orthognathic treatment with autotransplantation of impacted maxillary third molar. AB - This article presents orthognathic treatment in successful collaboration with autotransplantation of the maxillary third molar. The case report describes a 24 year-old man with severe mandibular protrusion and severe crowding in both dentitions. Overbite was 0 mm, overjet, -15 mm. Maxillary second molars and mandibular third molars were extracted, and presurgical multibracket treatment was begun. Maxillary third molars were impacted completely at 18 months in terms of presurgical tooth alignment. Autotransplantation of the teeth was achieved to substitute for extracted maxillary second molars. At 6 months after autotransplantation, the maxilla was advanced 4 mm on both sides through a Le Fort I procedure; left and right sides of the mandible were set back 15 and 18 mm, respectively, via sagittal split ramus osteotomy to improve mandibular protrusion. The total treatment period lasted 37 months. Autotransplantation treatment is an effective modality for tooth replacement when a donor tooth is available. Fully impacted maxillary third molars are potentially reliable candidates for autotransplantation. PMID- 19216596 TI - Mandibular rotation and remodeling changes during early childhood. AB - OBJECTIVE: To describe the mandibular rotation and remodeling of younger children. MATERIALS AND METHODS: The sample included 43 males and 43 females who participated in the Bolton-Brush Growth Study at Case Western Reserve University in Ohio. They were chosen on the basis of having Class I (n = 45) or Class II (n = 41) molar relationships and longitudinal lateral cephalograms at three developmental stages of the dentition: late primary (T1: 5.7 +/- 0.5 y), early mixed (T2: 8.4 +/- 0.6 y), and full permanent dentition (T3: 15.4 +/- 0.5 y). Each subject's cephalograms were traced and four landmarks were digitized. Cranial base and mandibular superimpositions were performed with the use of natural reference structures. RESULTS: Yearly rates of true rotation, apparent rotation, and angular remodeling showed significant (P < .05) changes throughout. True rotation was moderately correlated with angular remodeling and apparent rotation. Although no significant sex differences in annual rates of rotation were noted, subjects with Class I molar relationships showed significantly more angular remodeling from T2-T3 than did subjects with Class II molar relationships. Rates of true forward rotation were significantly greater with T1 T2 than with T2-T3 (1.3 and 0.7 degrees/y, respectively). CONCLUSION: Although significant amounts of true mandibular rotation and angular remodeling occur during childhood and adolescence, true rotation is greatest during the transition from late primary to early mixed dentition. PMID- 19216597 TI - Effectiveness of pit and fissure sealants in reducing white spot lesions during orthodontic treatment. A pilot study. AB - OBJECTIVE: A pilot investigation was performed to test the null hypothesis that highly filled (58%) resin sealants do not prevent white spot lesions in patients undergoing active orthodontic treatment. MATERIALS AND METHODS: A split-mouth design was applied to 60 healthy patients, with the sealant randomly allocated to either the right or the left side of each jaw. The sealant was applied to the incisors and canines from the gingival surface of the bracket to the free gingival margin. The contralateral teeth had the same type of bracket with no sealant. Sealants were placed on the experimental teeth 2 weeks to 3 months after initial bonding and were removed after 15 to 18 months. Intraoral photographs, visual assessments, and DIAGNOdent (KaVo Dental Corporation, Lake Zurich, Ill) measurements were used to assess white spot lesions after sealant removal. RESULTS: Six lesions on the teeth with sealants were identified visually, compared with 22 lesions on the teeth without sealants. The teeth without sealants had 3.8 times the number of white spot lesions than were noted on the sealed teeth. These sealants showed no visible signs of discoloration. The DIAGNOdent measured statistically significant differences between sealed and unsealed teeth in the maxilla (P < .001) and in the mandible (P = .010). DIAGNOdent measurements also showed a difference between sealed and unsealed teeth after the 28 teeth with visible lesions were excluded. CONCLUSION: The hypothesis was rejected. Ultraseal XT Plus clear sealant (Ultradent Products, South Jordon, Utah) produced a significant reduction in enamel demineralization during fixed orthodontic treatment and should be considered for use by clinicians to minimize white spot lesions. PMID- 19216598 TI - Craniofacial morphology in prematurely born children. AB - OBJECTIVE: To test the null hypothesis that there is no difference between the craniofacial morphology of prematurely born children and that of matched full term born controls. MATERIALS AND METHODS: White children 8 to 10 years of age, born at the University Hospitals of Lund and Malmo and living in the same part of Sweden, were included. One group consisted of 36 very preterm children, born during gestational weeks 29 to 32; the other group included 36 extremely preterm children, who were born before the 29th gestational week. Subjects were compared with a control group of 31 full-term children, who were matched for gender, age, nationality, and living area. One lateral head radiograph was taken for each child, and the cephalometric analysis included 15 angular and 11 linear variables. Also, the height, weight, and head circumference of each child were registered. RESULTS: A significantly shorter anterior cranial base and a less convex skeletal profile were found among extremely preterm children, and significantly shorter maxillary length was noted in both extremely preterm and very preterm groups as compared with full-term children. The lower incisors were significantly more retroclined and retruded in the extremely preterm group compared with the very preterm group and the full-term control group. Extremely preterm children were significantly shorter, and both extremely preterm and very preterm children had significantly lower weight and smaller head circumference compared with full-term children. CONCLUSION: The null hypothesis was rejected because several craniofacial parameters differed significantly between preterm and full-term born control children. PMID- 19216599 TI - Lip morphological changes in orthodontic treatment. Class II division 1: malocclusion and normal occlusion at rest and on smiling. AB - OBJECTIVE: To evaluate the morphological changes in the lips and to determine the degree of improvement in the smile after orthodontic treatment for Class II division 1 malocclusion. MATERIALS AND METHODS: The sample subjects were divided into two groups: a group that consisted of 20 adult female patients with Angle Class II division 1 malocclusion and a control group that consisted of 28 adult female volunteers with normal occlusion. Frontal photographs were taken before and after orthodontic treatment, and 35 landmarks were placed on each tracing made from the photograph. Thereafter, landmarks were digitized into an x- and y coordinate system with the subnasal point as the origin. The comparisons between pretreatment and posttreatment at rest and on smiling, and the comparisons between Class II division 1 and control group were made using Student's t-test. RESULTS: Both the upper and lower lips in the smile of patients in the Class II division 1 pretreatment group were positioned downward, and the upward movement of the upper lip and mouth corners was smaller in comparison with the control group. These characteristics of the Class II smile improved after orthodontic treatment, but the differences with the control group remained unchanged immediately after treatment. CONCLUSION: The soft tissue morphology shows a relative improvement after orthodontic treatment. PMID- 19216600 TI - Lip posture following debonding of labial appliances based on conventional profile photographs. AB - OBJECTIVE: To assess changes in lip posture following debonding of labial appliances on the basis of a new soft tissue analysis performed by computerized tools with the use of conventional nonstandardized profile photographic pictures. MATERIALS AND METHODS: Profile photographs of 33 patients were taken just before and just after debonding of labial brackets. Pictures were examined twice through custom-made analysis in which Viewbox software was used. The validity of this new analysis was assessed in a preliminary study. A reference line was constructed between the center of the tragus and the outer canthus. The point where this elongated line intersected with the profile was coined nasion-modified and served as the vertex point for angles used to evaluate the prominence of the lips. The final position of the lines was determined when they were tangent to the appropriate landmarks. Analysis of variance (ANOVA) and t-tests for paired differences were used. RESULTS: The reproducibility of this photoanalysis was confirmed in the preliminary study, in that no significant differences were found between measurements (P < .05). No statistically significant differences were noted in lip posture between pre-debonding and post-debonding pictures (P < .05). CONCLUSIONS: In keeping with the design of this study, soft tissue profile photoanalysis showed that the presence of bonded labial appliances had no effect on lip posture. PMID- 19216601 TI - Surgical accuracy of maxillary repositioning according to type of surgical movement in two-jaw surgery. AB - OBJECTIVE: To compare the surgical accuracy of the maxillary repositioning according to the maxillary surgical movement type (SMT) in two-jaw orthognathic surgery (TJOS). MATERIALS AND METHODS: The samples consisted of 52 Korean young adult patients with skeletal Class III malocclusion treated with TJOS by one surgeon. Lateral cephalograms were taken 1 month before (T0) and 1 day after surgery (T1). The samples were allocated into maxillary advancement (MA), total setback (MS), impaction (MI), and elongation (ME) according to SMT. The distance from the upper incisor tip and the mesiobuccal cusp tip of the upper first molar to the horizontal and vertical reference lines at T0 and T1 were measured. Any discrepancy between the surgical treatment objective (STO) and the surgical result less than 1 mm was regarded as accurate. The accuracy rate (AR [number of the accurate sample/number of the sample] x 1000) and the surgical achievement ratio (SAR [amount of movement in surgical result/amount of movement in STO] x100) were calculated. Analysis variance (ANOVA) and crosstab analyses were used for statistical analysis. RESULTS: Although the MS (69.2%) and MI (69.0%) showed a lower AR than the MA (87.5%) and ME (83.3%), there was no significant difference in the distribution of accurate and inaccurate samples among the groups. The mean discrepancy between the STO and the surgical result was less than 1 mm in all groups. Although the ME (93.54%) showed a tendency of undercorrection and the MS (107.10%) and MI (105.42%) a tendency of overcorrection, there was no significant difference in SAR among the groups. CONCLUSIONS: If the surgical plan and procedure is done with caution, the MS and MI can be regarded as just as accurate a procedure as the MA and ME. PMID- 19216602 TI - Posterior crossbite and functional changes. A systematic review. AB - OBJECTIVE: To assess, by systematically reviewing the literature, the functional changes of the masticatory muscles associated with posterior crossbite in the primary and mixed dentition. MATERIALS AND METHODS: A literature survey from the Medline database covering the period from January 1965 to February 2008 was performed. Randomized controlled trials, controlled clinical trials, and clinical trials that evaluated bite force, surface electromyography, and signs and symptoms of temporomandibular disorders (TMD) were included. Two reviewers extracted the data independently and assessed the quality of the studies. RESULTS: The search strategy resulted in 494 articles, of which 8 met the inclusion criteria. Children with posterior crossbite can have reduced bite force and asymmetrical muscle function during chewing or clenching, in which the anterior temporalis is more active and the masseter less active on the crossbite side than the noncrossbite side. Moreover, there is a significant association between posterior crossbite and TMD symptomatology. CONCLUSION: The consequences of the functional changes for the growth and development of the stomatognathic system deserves further investigation. PMID- 19216603 TI - Opening of circumaxillary sutures by alternate rapid maxillary expansions and constrictions. AB - OBJECTIVE: To analyze quantitatively the circumaxillary suture opening after alternate rapid maxillary expansions and constrictions (Alt-RAMEC). MATERIALS AND METHODS: Twelve inbred cats were randomly grouped into two equal groups for 1 week of rapid maxillary expansion (RME) (1 mm/day) or 5 weeks of Alt-RAMEC (1 mm/day). At the end of the experiment, the craniofacial skeleton of each cat was harvested. Each circumaxillary suture was then probed at three sites with a 0.5 mm pointed periodontal probe. A smooth probing without penetration was an ineffective suture opening (<0.5 mm), while a probing with penetration was an effective suture opening (>0.5 mm). For each suture, the quantity of suture opening (%) was the effective suture opening/(effective + ineffective suture opening). The intergroup differences were analyzed by chi-square test (P < .05). RESULTS: Five weeks of Alt-RAMEC opened the circumaxillary sutures significantly more than 1 week of RME. This affected the circumaxillary sutures running coronally and articulating directly to the maxilla (56.9% vs 36.1%, P < .001), the sutures running sagittally, but articulating indirectly to the maxilla (94.4% vs 64.8%, P < .001), and the sutures running coronally, but articulating indirectly to the maxilla (58.3% vs 33.3%, P < .01). The sutures running sagittally were opened significantly more (94.4%-100.0%) than those running coronally (56.9%-58.3%), no matter if they articulated directly or indirectly with the maxilla. CONCLUSIONS: Alt-RAMEC opens both the sagittally and coronally running circumaxillary sutures quantitatively more than conventional RME. However, more than 5 weeks of Alt-RAMEC would be needed to increase the opening of the coronally running circumaxillary sutures. ( PMID- 19216604 TI - Osteoprotegerin and ligand of receptor activator of nuclear factor kappaB expression in ovariectomized rats during tooth movement. AB - OBJECTIVE: To test the null hypothesis that increased tooth displacement in ovariectomized rats is not related to differential expressions of OPG and RANKL in the periodontium. MATERIALS AND METHODS: Eighty-four 12-week female rats were used; half were ovariectomized and half were not. Three months later, the maxillary first molar was moved mesially. Groups of rats were sacrificed at days 0, 1, 3, 5, 7, 10, and 14 after activation. Tooth movement was measured at each time point. OPG and RANKL expressions were examined through immunohistochemistry. RESULTS: Ovariectomized and nonovariectomized rats showed three-phase tooth movement. In both groups, OPG expression increased at the tension area and RANKL increased at the pressure area. The OPG/RANKL ratio coincided with tooth movement, especially in the linear phase from 7 to 14 days. CONCLUSIONS: The null hypothesis is rejected. The increased rate of tooth movement in ovariectomized rats was related to differential expressions of OPG and RANKL. PMID- 19216605 TI - Stability in dental changes in RME and SARME: a 2-year follow-up. AB - OBJECTIVE: To compare the effects of rapid maxillary expansion (RME) and surgically assisted rapid maxillary expansion (SARME) on dentoalveolar structures following orthodontic treatment, as well as stability at 2-year follow-up. MATERIALS AND METHODS: Two groups of subjects were used in the study. Group 1 consisted of 14 subjects (mean age, 12.7 +/- 1.4 years) who were treated with RME, and Group 2 consisted of 13 subjects (mean age, 18.5 +/- 2.3 years) who were treated with SARME. In both groups, all cases had a maxillary width deficiency with bilateral crossbites. Maxillary dental casts were available at three different intervals: pretreatment (T1), after orthodontic treatment (T2), and at follow-up recall (T3). Intermolar and interpremolar width, palatal height, and maxillary arch depth and length were assessed from maxillary dental casts. RESULTS: Treatment by RME and SARME produced significant increases in intermolar and interpremolar width and maxillary arch length after expansion (T2) (P < .05). The amount of relapse was not significantly different 2 years after treatment (P > .05). CONCLUSIONS: Although age ranges of the patient groups are different, the dentoalveolar responses of RME and SARME were similar after orthodontic treatment. PMID- 19216606 TI - Effects of different chlorhexidine formulations on shear bond strengths of orthodontic brackets. AB - OBJECTIVE: To test the hypothesis that the application of different chlorhexidine formulations to the etched enamel will not affect shear bond strength (SBS). MATERIALS AND METHODS: Forty-four freshly extracted human premolars were collected and stored in distilled water. The teeth were etched with 37% phosphoric and were rinsed and dried. The teeth then were divided into four equal groups. While Group 1 served as a control, Groups 2 to 4 were treated before bonding with a chlorhexidine formulation that included solution (2%), gel (1%), and mouthwash (0.2%). Orthodontic brackets were bonded with Transbond XT (3M Unitek, Monrovia, Calif). Bond strength results were evaluated with the use of one-way analysis of variance (ANOVA) (P < .05) and post hoc tests. Modes of failures were verified by means of scanning electron microscopy. RESULTS: Although no statistically significant difference was observed between Groups 1 and 4 (P > .05), both were statistically superior to Groups 2 and 3 (P < .05). In this in vitro study, the observed measures for Groups 2 and 3 (14.5-10.6 MPa) were lower than those for Groups 1 and 4 (27.3-24.9 MPa), but these values were much higher than those required for clinical use (6-8 MPa). CONCLUSION: The hypothesis is rejected. The application of chlorhexidine mouth rinse before bonding had no significant effect on the SBS value, and the application of chlorhexidine solution and gel significantly decreased SBS. PMID- 19216607 TI - Chromium release from new stainless steel, recycled and nickel-free orthodontic brackets. AB - OBJECTIVE: To test the hypothesis that there is no difference in the amounts of chromium released from new stainless steel brackets, recycled stainless steel brackets, and nickel-free (Ni-free) orthodontic brackets. MATERIALS AND METHODS: This in vitro study was performed using a classic batch procedure by immersion of the samples in artificial saliva at various acidities (pH 4.2, 6.5, and 7.6) over an extended time interval (t(1) = 0.25 h, t(2) = 1 h, t(3) = 24 h, t(4) = 48 h, t(5) = 120 h). The amount of chromium release was determined using an atomic absorption spectrophotometer and an inductively coupled plasma atomic emission spectrometer. Statistical analysis included a linear regression model for repeated measures, with calculation of Huber-White robust standard errors to account for intrabracket correlation of data. For post hoc comparisons the Bonferroni correction was applied. RESULTS: The greatest amount of chromium was released from new stainless steel brackets (0.52 +/- 1.083 microg/g), whereas the recycled brackets released 0.27 +/- 0.38 microg/g. The smallest release was measured with Ni-free brackets (0.21 +/- 0.51 microg/g). The difference between recycled brackets and Ni-free brackets was not statistically significant (P = .13). For all brackets, the greatest release (P = .000) was measured at pH 4.2, and a significant increase was reported between all time intervals (P < .002). CONCLUSION: The hypothesis is rejected, but the amount of chromium released in all test solutions was well below the daily dietary intake level. PMID- 19216608 TI - Marfan syndrome-an orthodontic perspective. AB - Marfan syndrome is a heritable disorder of connective tissue that can affect the heart, blood vessels, lungs, eyes, bones, and ligaments. It is characterized by tall stature, elongated extremities, scoliosis, and a protruded or caved-in breastbone. Patients typically have a long, narrow face. A high-arched palate produced by a narrow maxilla and skeletal Class II malocclusion due to mandibular retrognathia are other common features. For a patient with no family history of the disorder, at least three body systems must be affected before a diagnosis can be made. Individuals affected by the syndrome routinely seek orthodontic treatment to correct the orofacial manifestations. In this report, the authors present the records of three patients with Marfan syndrome who were treated at a dental school. Two patients had severe periodontal disease in the absence of significant contributing local factors. The presentation of systemic symptoms and typical physical characteristics varied. The syndrome thus went unnoticed in one patient for many years. We discuss here the observed intraoral findings and the progress of orthodontic treatment to provide a brief overview of the challenges involved in treating such patients. PMID- 19216609 TI - In vitro antibacterial and cytotoxicity assessments of an orthodontic bonding agent containing benzalkonium chloride. AB - OBJECTIVE: To assess the antibacterial activity and cytotoxicity of an orthodontic bonding material containing an antibacterial agent. MATERIALS AND METHODS: Superbond C&B (4-methacryloxyethyl trimellitate anhydride/methyl methacrylate-tri-n-butyl borane [4-META/MMA-TBB]) resin was mixed with benzalkonium chloride (BAC) to obtain final BAC concentrations of 0.25%, 0.75%, 1.25%, 1.75%, 2.5%, and 5.0% (wt/ wt). Antibacterial activity against Streptococcus mutans and Streptococcus sobrinus was evaluated by soaking the BAC resin in distilled water at 37 degrees C for periods of 30, 90, and 180 days. Antibacterial activity of the BAC-resin was measured by the disk diffusion method, and the inhibition zone around each sample was measured and recorded. For evaluation of cytotoxicity, BAC-resin samples were put into cell culture inserts placed above human gingival cells and were incubated at 37 degrees C for 1, 3, and 6 days. Cytotoxicity was assessed with a tetrazolium bromide reduction assay. RESULTS: The antibacterial activity of BAC-incorporated resin samples decreased significantly after immersion in water for 180 days, regardless of BAC concentration. The antibacterial activity of nonimmersed resin containing 0.25% or 1.75% BAC was comparable with that of 5.0% BAC-resin immersed for 180 days. In cytotoxicity tests, most cells died when exposed to resins containing 1.75%, 2.5%, and 5% BAC. No difference was observed between resins containing 0.25% and 0.75% BAC at 1, 3, and 6 days of culture. CONCLUSIONS: The addition of BAC to 4 META/MMA-TBB resin confers an antibacterial effect even after immersion in water, and 4-META/MMA-TBB resin containing 0.25% to 0.75% BAC has no significant cytotoxic effect. PMID- 19216610 TI - Holographic analysis of the initial canine displacement produced by four different retraction springs. AB - OBJECTIVES: To estimate and compare the magnitude and direction of initial displacement of the canine produced by four different canine retraction springs. MATERIALS AND METHODS: A freshly macerated mandible was used as an experimental model. First premolars were extracted, and strap-up was done with a 0.018 inch Roth Preadjusted Edgewise System (3M Unitek, Monrovia, Calif). The canine was subjected to loads of 4 oz, 5 oz, and 6 oz, each applied by four different canine retraction springs, that is, closed coil spring, open coil spring, PG spring, and T-loop retraction spring. The magnitude and direction of the initial displacement of the canine were studied by means of double-exposure interferometry. RESULTS: The PG spring produced the highest initial displacement for a given force among the force systems used, followed by open coil, closed coil, and T-loop. Maximum tipping was observed with the open coil spring, followed by the PG spring, the closed coil spring, and the T-spring. CONCLUSION: The T-loop may be preferred whenever minimal tipping is performed. The PG spring may be preferred over other springs whenever a higher magnitude of displacement is desired. Closed coil springs may be preferred whenever a reasonable magnitude of displacement is required and reasonable tipping is allowed. PMID- 19216611 TI - Class II correction with the Cantilever Bite Jumper. AB - OBJECTIVE: To identify the skeletal, dentoalveolar, and soft tissue changes that occur during Class II correction with the Cantilever Bite Jumper (CBJ). MATERIALS AND METHODS: This prospective cephalometric study was conducted on 26 subjects with Class II division 1 malocclusion treated with the CBJ appliance. A comparison was made with 26 untreated subjects with Class II malocclusion. Lateral head films from before and after CBJ therapy were analyzed through conventional cephalometric and Johnston analyses. RESULTS: Class II correction was accomplished by means of 2.9 mm apical base change, 1.5 mm distal movement of the maxillary molars, and 1.1 mm mesial movement of the mandibular molars. The CBJ exhibited good control of the vertical dimension. The main side effect of the CBJ is that the vertical force vectors of the telescope act as lever arms and can produce mesial tipping of the mandibular molars. CONCLUSIONS: The Cantilever Bite Jumper corrects Class II malocclusions with similar percentages of skeletal and dentoalveolar effects. PMID- 19216612 TI - Stability of mini-screws invading the dental roots and their impact on the paradental tissues in beagles. AB - OBJECTIVE: To examine the stability of mini-screws that invade a dental root by measuring the retention period/failure rate, and to illustrate their effects on paradental tissues. MATERIALS AND METHODS: Three adult male beagle dogs received 48 orthodontic mini-screws. Half of the mini-screws were implanted to invade the roots, and the rest were placed in the middle of the alveolar bone. Half of the mini-screws were loaded immediately. The retention period of the mini-screws was documented. The dogs were euthanized after 8 weeks, and tissue samples were examined histologically. RESULTS: The failure rate of the mini-screws that invaded the roots was 79.2%, and that of the mini-screws in the middle of the alveolar bone was 8.3%. The application of force had little effect on the failed mini-screws. Moderately injured roots were repaired with osteoid and/or cementoid tissues with normal periodontal ligaments, followed by recovery of the original configuration. CONCLUSION: Orthodontic mini-screws had a higher failure rate when placed to invade the dental roots. However, minimally damaged dental roots do not adversely affect the healing process. PMID- 19216613 TI - Orthodontic treatment including autotransplantation of a mature tooth. AB - The patient was a 24-year-old Japanese female. The chief complaints were crowding and masticatory dysfunction due to the missing right first molar. Her maxillary first premolars had been extracted when she was a primary school student. We planned orthodontic treatment with extraction of the mandibular first premolars and transplantation of the mandibular left first premolar into the maxillary right first molar area. We made a diagnostic setup model to initiate an appropriate treatment plan for the discrepancy in tooth size ratio. Following the diagnostic setup model, the space in the maxillary right first molar area was closed by a small amount of tooth movement, and a good occlusion was achieved. The patient had been in retention for 7 years, and the occlusion has been maintained very well during this time. In the follow-up, 10 years after autotransplantation, no signs of inflammatory or replacement root resorption were found, and marginal bone support appeared similar to that of neighboring teeth. PMID- 19216616 TI - Helminths from an introduced species (Tupinambis merianae), and two endemic species (Trachylepis atlantica and Amphisbaena ridley) from Fernando de Noronha Archipelago, Brazil. AB - The present study reports the occurrence of helminths in the introduced species Tupinambis merianae (tegu lizard), and in two endemic species Trachylepis atlantica (small lizard) and Amphisbaena ridleyi (two-head-snake lizard ), from Fernando de Noronha Archipelago, State of Pernambuco, Brazil. Nine species of helminths were found, mainly in the digestive tract and accessory organs, with the following prevalence (P) and mean infection intensity (MII). Tupinambis merianae: Diaphanocephalus galeatus (P = 96%, MII = 20.5), Spinicauda spinicauda (P = 100%, MII = 197.8), and Oochoristica sp.l (P = 20%, MII = 4.4). Trachylepis atlantica: Moaciria alvarengai (P = 20%, MII = 1.4), S. spinicauda (P = 92%, MII = 22.1), Mesocoelium monas (P = 4%, MII = 3.0), Platynosomum sp. (P = 8%, MII = 7.0), and Oochoristica sp.2 (P = 16%, MII = 1.25). Amphisbaena ridleyi: Aplectana albae (P = 96%, MII = 143.4), Thelandros alvarengai (P = 4%, MII = 1.0), Me. monas (P = 44%, MII = 2.8), Platynosomum sp. (P = 36%; MII = 13.8), and Oochoristica sp.2 (P = 48%; MII = 2.17). More than 80% of T. merianae were infected with 2, or more, helminth species. In Tr. atlantica, single-species infections were present in 50% of the specimens, but co-occurrence of 2 parasites was also high (41.7%). In A. ridleyi, multiple infections were more common, with up to 5 parasite species present. The helminth fauna observed allowed us to conclude that helminths can be carried together with their host when they colonize new geographic localities and that these introduced helminths can, in turn, colonize endemic, or native, hosts. PMID- 19216617 TI - Integrated analysis of safety and efficacy of a plasma- and albumin-free recombinant factor VIII (rAHF-PFM) from six clinical studies in patients with hemophilia A. AB - BACKGROUND: Hemophilia A is an X-linked bleeding disorder that results from insufficient levels of factor VIII (FVIII) coagulant activity. OBJECTIVE: To evaluate the efficacy and safety of ADVATE rAHF-PFM (Baxter Healthcare Corporation), a recombinant FVIII concentrate manufactured without human or bovine blood-derived additives, and to assess the effect of compliance with prophylactic use in preventing bleeding episodes (BEs). METHODS: Clinical data were integrated from six prospective studies. Two hundred thirty-four hemophilia A subjects (FVIII levels < or = 2%) (median age 14.7 (range: 0.02 - 72.7) years) were included. RESULTS: BEs were managed with one or two infusions and nearly all (1953/1956) responded to treatment. Compliance with a prophylactic treatment regimen significantly reduced the incidence of BEs (p = 0.0061) and prevented non traumatic joint BEs (median annualized BE rate was 0). One previously treated subject developed an inhibitor; no other safety concerns were observed. CONCLUSIONS: These results reinforce the efficacy and safety of rAHF-PFM and suggest that compliance is an essential contributor to the effectiveness of prophylaxis in the treatment of hemophilia A. PMID- 19216618 TI - Antigen-antibody immunogenic complex: promising novel vaccines for microbial persistent infections. AB - BACKGROUND: Persistent microbial infections are major public health problems worldwide. Immunotherapies have become an important treatment for persistent infections. With the increasing senescent population, low responsiveness to the current preventive vaccines is another challenge for control of infectious diseases. OBJECTIVE/METHODS: Active immunotherapy by antigen-antibody complexes (IC) is reviewed. RESULTS/CONCLUSIONS: IC have shown effects in an hepatitis B surface antigen positive transgenic mouse model by reducing HBsAg, inducing anti HBs and initiating cytolytic responses. Phase I, IIA and IIB clinical trials in around 300 viral hepatitis B patients have shown promising results. The mechanisms of IC are mainly modulation of antigen uptake and antigen processing and antigen presentation by IC. The prospects for employing IC in treatment of other microbial persistent infections and for prevention in immunocompromized individuals are discussed. PMID- 19216620 TI - Supramolecular approaches to biological therapy. AB - Supramolecular chemistry is a useful methodology for construction of nano- or micro-sized objects and can significantly contribute to nanotechnology through so called bottom-up processing. In addition, supramolecular self-assembled structures can mimic some aspects of biological systems. Bio-related functions such as molecular sensing, controlled release, signaling and materials separations have been realized. Supramolecular chemistry is a multidisciplinary field that includes subjects such as molecular design and nanosized materials. In this article recent examples of supramolecular chemistry in the context of biological therapy are introduced and classified into five categories: small supramolecular systems; designer polymers; self-assembled structures; predesigned assemblies; and nanomaterials. Finally, hierarchic organization of supramolecular structures for advanced functions is introduced to illustrate future directions of investigation. We hope that scientists studying therapeutic applications receive inspiration from this review to exploit the opportunities offered by supramolecular chemistry in their respective research areas. PMID- 19216621 TI - Telomerase-specific virotherapy for human squamous cell carcinoma. AB - BACKGROUND: Replication-selective tumor-specific viruses present a novel approach for treatment of neoplastic disease. They are designed to induce lysis after propagation within the tumor. Human telomerase is active in over 85% of primary cancers and its activity correlates closely with human telomerase reverse transcriptase (hTERT) expression. OBJECTIVES: Oncolytic viruses, Telomelysin and TelomeScan, that combine the specificity of hTERT promoter-based expression systems with the lytic efficacy of replicative viruses were developed. The goal was to confirm the efficacy of the viruses for human squamous cell carcinoma. RESULTS/CONCLUSION: Squamous cell carcinoma of the head and neck (SCCHN) is characterized by locoregional spread, and is clinically accessible, making it an attractive target for intratumoral virotherapy. The viruses replicated efficiently and induced killing in a panel of human cancer cell lines including SCCHN cells in vitro and in vivo. These results illustrate the potential of telomerase-specific oncolytic viruses for treatment of human SCCHN. PMID- 19216619 TI - In vivo tracking of cellular therapeutics using magnetic resonance imaging. AB - BACKGROUND: The success of many cell-based therapies is highly dependent on the accurate delivery, dosing and trafficking of the cellular therapeutic. In vivo magnetic resonance (MR) cell tracking provides a means to non-invasively and longitudinally evaluate these parameters for cellular therapy. OBJECTIVE: To provide an overview of MR cell tracking and how cellular therapeutics might be improved by utilizing this technology. METHODS: We focused on the technologies utilized for stem cell and immunotherapies in preclinical models of disease. RESULTS/CONCLUSION: New technologies in MR cell tracking will soon take the field beyond preclinical studies and begin to show benefits in clinical trials of novel experimental cell-based therapies. PMID- 19216622 TI - Countering tumor-induced immunosuppression during immunotherapy for pancreatic cancer. AB - BACKGROUND: Vaccines for pancreatic cancer have been challenged by a number of factors, especially the immunosuppressive microenvironment within the tumor that allows for escape from immune surveillance. OBJECTIVE/METHODS: We sought to identify results that define mechanisms of pancreatic-cancer-associated immunosuppression and strategies that might be useful to overcome them thereby resulting in effective immune responses to cancer vaccines capable of deleting pancreatic cancer cells. RESULTS/CONCLUSION: Immunosuppressive tumor-associated macrophages (TAMs), myeloid-derived suppressor cells (MDSC), and regulatory T cells (Treg) reside in tumors, and their products along with tumor derived products (such as VEGF, TGFbeta and IL-10), create a microenvironment that counters immune activation and attack. Immunotherapy with cancer vaccines must include strategies to modulate these immunosuppressive cell types and tumor byproducts. Clinical trials are beginning to test these strategies. PMID- 19216623 TI - Monoclonal antibody therapies for solid tumors. AB - BACKGROUND: About 1.4 million new cancer cases are expected to be diagnosed in the US in 2008 and the overall costs of cancer treatment is estimated at > $200 billion per year. Over half a million people will die from cancer each year. Monoclonal antibodies have attracted attention because of their potential clinical benefits. Currently, there are four monoclonal antibodies that have been approved by the FDA for the treatment of solid tumor malignancies. Over 100 other antibodies are in early- and late-phase clinical testing. OBJECTIVES/METHODS: Clinical efficacy of FDA-approved and investigational monoclonal antibody therapies are summarized. RESULTS/CONCLUSIONS: Anti-tumor responses have been modest. However, the advent of novel antibody engineering and combination therapy strategies has generated much optimism for current and future antibody-based therapeutics in solid tumors. PMID- 19216625 TI - Insulin glulisine: aspects of basal/bolus therapy for optimized treatment of diabetes mellitus. AB - BACKGROUND/OBJECTIVE: The prevalence of diabetes mellitus continues to increase throughout the world with an expected doubling of documented cases between 2006 and 2030, diabetes deaths are expected to increase by more than 50% from 2006 to 2015. Therapeutic interventions should be based on scientific evidence, including earlier insulin initiation and use of a basal-bolus approach that better mimics normal physiology. METHODS: This review focuses on insulin glulisine in the basal bolus approach. It details pharmacological data, clinical efficacy, safety and tolerability, and potential effects on current treatment regimens. RESULTS/CONCLUSION: Insulin glulisine offers a more rapid onset of action and shorter duration of action compared with regular human insulin. These characteristics result in an action profile that closely mimics the normal postmeal endogenous insulin response, thus fulfilling the prandial insulin requirement and making the basal-bolus treatment approach clinically achieveable. PMID- 19216626 TI - Pollinex Quattro Tree: allergy vaccine. AB - An overview of short-term specific immunotherapy (ST-SIT) highlighting Pollinex Quattro (PQ) Tree is presented. The product development of this novel allergy vaccine using modifying agent glutaraldehyde, adjuvant monophosphoryl lipid and L tyrosine have heralded a superior ST-SIT. Since 1999 when PQ was founded in Germany, various research trials assessing both the standardization and clinical studies have been done. A review of these studies demonstrates the efficacy and safety of PQ Tree in both pediatric and adult trials. The uniqueness of this product allows a shorter course of four pre-seasonal injections to provide control of allergy symptomatology in seasonal rhinitis patients. The PQ Tree product studies show a similar efficacy and safety profile to the grass formulation trial. PMID- 19216624 TI - Potent human monoclonal antibodies against SARS CoV, Nipah and Hendra viruses. AB - BACKGROUND: Recently, several potently neutralizing fully human monoclonal antibodies (hmAbs) targeting the severe acute respiratory syndrome-associated coronavirus (SARS CoV) S glycoprotein, and the G glycoprotein of the paramyxoviruses Hendra virus (HeV) and Nipah virus (NiV) have been discovered [corrected]. OBJECTIVE: To examine, compare and contrast the functional characteristics of hmAbs with the potential for prophylaxis and treatment of diseases caused by SARS CoV, HeV and NiV. METHODS: A review of relevant literature. RESULTS/CONCLUSIONS: Structural, functional and biochemical analyses [corrected] have provided insights into the molecular mechanisms of receptor recognition and antibody neutralization, and suggested that these antibodies alone or in combination could fight the viruses' heterogeneity and mutability, which is a major problem in the development of effective therapeutic agents against viruses, including therapeutic antibodies. PMID- 19216627 TI - Oncolytic virus as an agent for the treatment of malignant ascites. AB - Oncolytic virus H101 is an adenovirus with an E1B-55KD gene deletion, which selectively replicates in tumor cells, rather than in normal cells, resulting in specific tumor cytolysis. Taking the selective killing of peritoneally planted tumor cells by the oncolytic virus H101 as the rationale for the ascites treatment, we report in this paper the intraperitoneal H101 in 9 patients with malignant ascites. The interval between the paracentesises increased from a mean of 12.4+/- 3.4 to 39.9 +/- 11.6 days (p < 0.001). The 30-day response was classified into complete response (CR) (no fluid recurrence), partial response (PR) (fluid recurrence <50%), and no response (NR) (fluid recurrence > or =50% or requiring paracentesis). Nine (9) patients had 3 CRs, 2 PRs, and 4 NRs. Intraperitoneal H101 was well tolerated, and no severe adverse effects were observed. Further, this report also highlights the potential of H101 in malignant ascites treatment. PMID- 19216628 TI - Rituximab and chemotherapy in primary gastric lymphoma. AB - PURPOSE: We performed a phase II clinical trial to assess the efficacy and toxicity of the addition of rituximab and conventional chemotherapy in primary gastric lymphoma (PGL). METHODS: Forty-two (42) patients with PGL, stage IE and IIE, and with low- or low-intermediate clinical risk were treated in a prospective longitudinal study with standard CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone) chemotherapy and rituximab (375 mg/m2, intravenously) on day 1 of each cycle administered every 21 days, for 6 cycles. The endpoint was to assess improvement in outcome measured by prolongation in event-free survival (EFS) and overall survival (OS). Complete response was achieved in 40 cases (95%) (95% confidence interval [CI]: 88%-102%). Relapse was observed in 2 cases. Two (2) patients died secondary to tumor progression. Thus, actuarial 5-year EFS was 95% (95 % CI: 87%-104%) and OS was 95% (95% CI: 88% 101%), which was not statistically different to historic controls. Acute toxicity was minimal and well tolerated, 4 cases developed late toxicity, 2 cases of herpes zoster infection, and 2 cases with granulocytopenia; in 1 case, the patient continued with mild granulocytopenia 3 years after treatment. CONCLUSIONS: The addition of rituximab to CHOP chemotherapy did not improve outcome in early-stage PGL. PMID- 19216629 TI - Real-time PCR analysis of micro-RNA expression in ionizing radiation-treated cells. AB - The cellular response to ionizing radiation exposure is very complex and involves many pathways. A wide variety of biologic effects are induced after exposure to ionizing radiation, ranging from DNA damage processing, signal transduction, mutations, altered gene expression, cell-cycle arrest, genomic instability, and induction of carcinogenesis to cell death. To gain insight into this complex response, global alterations in the expression of genes in irradiated cells have been examined. Recent studies have provided evidence to associate micro-RNA (miRNA) with many cellular processes, including carcinogenesis, timing of cell fate decision, apoptosis, and metabolic pathways controlling a range of events. The small noncoding miRNA are emerging as critical components in controlling the gene expression. Because miRNA target so many genes, we hypothesized that alterations in their expression may be associated with the overall response of cells to radiation treatment. To explore the role of miRNA in cellular response to ionizing radiation, we monitored the expression levels of several miRNA by employing the stem-loop real-time polymerase chain reaction in Jurkat and TK6 cells treated with gamma-radiation. The expression levels of several members of the let-7 family miRNA that functionally inhibit the mRNAs of Ras oncogenes were upregulated after ionizing radiation treatment in Jurkat cells but were downregulated in TK6 cells. The expressions of miRNA associated with MYC translocation were upregulated in both cell types. The modulation of miRNA involved in various cancers was also examined. These results provide a first glimpse to indicate the involvement of miRNA in radiation-induced stress response and will lead to functional studies dissecting the molecular details of these processes. PMID- 19216630 TI - 166Ho-labeled hydroxyapatite particles: a possible agent for liver cancer therapy. AB - PURPOSE: Intra-arterial administration of particulates labeled with -emitting radionuclides is one of the promising modalities for the treatment of liver cancer. 166Ho [T1/2=26.9 hours, E(max) = 1.85 MeV, Egamma = 81keV (6.4%)] could be envisioned as an attractive radionuclide for use in liver cancer therapy owing to its high energy emission, short half-life, and feasibility of its production with adequately high specific activity and radionuclidic purity. Hydroxyapatite (HA) particles in the size range of 20-60 micros were chosen as the particulate carrier due to their biocompatibility and ease of labeling with lanthanides. METHODS: 166Ho was produced by thermal neutron bombardment on a natural holmium target. HA particles of the desired size range were synthesized, characterized, and radiolabeled with 166Ho. The biologic behavior of166Ho-HA was tested in normal Wistar rats by carrying out biodistribution and imaging studies. RESULTS: 166Ho was produced with a specific activity of 5.55-6.48 TBq/g and radionuclidic purity of approximately 100%. HA particles were labeled with 166Ho with a high radiochemical purity of >99% and good in vitro stability up to 7 days. The biodistribution and imaging studies revealed satisfactory hepatic retention (approximately 89% of injected activity after 2 days) with insignificant uptake in any other major organ/tissue. CONCLUSIONS: 166Ho-HA exhibited promising features as an agent for liver cancer therapy in preliminary studies and warrants further investigation PMID- 19216631 TI - In-vitro internalization and in-vivo tumor uptake of anti-EGFR monoclonal antibody LA22 in A549 lung cancer cells and animal model. AB - PURPOSE: Internalization is one of the key steps for anticancer immunoconjugates to deliver the drugs inside of cancer cells. Herein, the internalization property of antiepidermal growth factor receptor (EGFR) monoclonal antibody (mAb) LA22 was evaluated. MATERIALS AND METHODS: The binding and internalization properties of LA22 on A549 cells were investigated by using 125I-LA22. In vitro internalization was also confirmed by indirect fluorescent staining. In vivo tumor targeting and internalization of 125I-LA22 were evaluated in the A549 nude mice model. RESULTS: The mAb LA22 showed a high affinity to EGFRs expressed on A549 cells (Kd = 0.69 +/- 0.13 nM). The in vitro internalization of LA22 was time- and temperature dependent. The cell-surface-bound LA22 was rapidly internalized at 37 degrees C. The experimental results of LA22 internalization obtained from radioassay and fluorescent staining were consistent with a good linear correlation. At 72 hours postinjection, a clear gamma-image of tumor was obtain in A549 tumor xenografts, and the tumor uptake of 125I-LA22 was 8.00 +/- 0.61 percent injected dose per gram (%ID/g) (2.19 +/- 0.37 %ID/g for 125I-mIgG). Similar to the in vitro observation, 64.06% of the cell-bound mAb LA22 was internalized into the tumor cells in vivo. CONCLUSIONS: The mAb, LA22, is a rapid, high-internalizing antibody, and this property makes it a promising vehicle for tumor-targeted drug delivery. PMID- 19216632 TI - Three-dimensional porous biodegradable polymeric scaffolds fabricated with biodegradable hydrogel porogens. AB - We have developed a new fabrication technique to create three-dimensional (3D) porous poly(epsilon-caprolactone fumarate) (PCLF) scaffolds using hydrogel microparticle porogens, as an alternative to overcome certain limitations of traditional scaffold fabrication techniques such as a salt leaching method. Both natural hydrogel, gelatin, and synthetic hydrogel, poly(ethylene glycol) sebacic acid diacrylate, were used as porogens to fabricate 3D porous PCLF scaffolds. Hydrogel microparticles were prepared by a single emulsion technique with the particle size in the range of 100-500 microm after equilibrium in water. The pore size distribution, porosity, pore interconnectivity, and spatial pore heterogeneity of the 3D PCLF scaffolds were assessed using micro-computed tomography and imaging analysis. Scaffolds fabricated with the hydrogel porogens had higher porosity and pore interconnectivity as well as more homogeneous spatial pore distribution, compared to the scaffolds made from the salt leaching process. Compressive moduli of the scaffolds were also measured and showed that lower porosity yielded greater modulus of the scaffolds. Overall, the new fabrication technology using hydrogel porogens may be beneficial for certain tissue engineering applications. PMID- 19216633 TI - In vivo factors influencing the freezing cycle during cryoablation of small renal masses. AB - PURPOSE: The aim of this study is to present a procedural analysis of the cryoablations performed in our department for small renal tumors and to try to identify clinical parameters or factors that influence the freezing rate during the procedure. PATIENTS AND METHODS: We collected all data from the procedures performed in our department until August 2007. Based on the intraoperative biopsy result, we grouped the cases in two groups: renal-cell carcinoma (RCC) and benign. We calculated the freezing rate in both groups and compared them. Finally, we performed a univariate and multivariate analysis to identify clinical parameters that significantly influence the freezing rate. RESULTS: A total of 70 cryoablations of small renal tumors in 67 patients were performed during this period. From these, 56 procedures met the inclusion criteria and were analyzed further. The RCC group consisted of 48 cases (39 RCC and 9 lesions with a nondiagnostic biopsy) while 8 formed the benign group. There was no difference in the freezing rate between these two groups. Preoperative creatinine levels above 120 IU, diabetes mellitus, American Society of Anesthesiologists score 3, and location of the tumor at the lower pole were found to increase the freezing rate. The only factor that significantly decreased the freezing rate was the presence of chronic obstructive pulmonary disease. The multivariate analysis showed that the location of the tumor and diabetes mellitus influence more significantly the temperature v time graph. CONCLUSIONS: The freezing rate during cryotherapy of small renal tumors is significantly influenced by various clinical factors, while there are no differences in the freezing rate of those proven small malignant tumors and the small benign lesions. PMID- 19216634 TI - Use of prolonged hypothermia to treat ischemic and hemorrhagic stroke. AB - Therapeutic (induced) hypothermia (TH) has been extensively studied as a means to reduce brain injury following global and focal cerebral ischemia, intracerebral hemorrhage (ICH), and subarachnoid hemorrhage (SAH). Here, we briefly review the clinical and experimental evidence supporting the use of TH in each condition. We emphasize the importance of systematically evaluating treatment parameters, especially the duration of cooling, in each condition. We contend that TH provides considerable protection after global and focal cerebral ischemia, especially when cooling is prolonged (e.g., >24 h). However, there is presently insufficient evidence to support the clinical use of TH for ICH and SAH. In any case, further animal work is needed to develop optimized protocols for treating cardiac arrest (global ischemia), and to maximize the likelihood of successful clinical translation in focal cerebral ischemia. PMID- 19216635 TI - Combination therapy with hypothermia for treatment of cerebral ischemia. AB - Mild hypothermia is an established neuroprotectant in the laboratory, showing remarkable and consistent effects across multiple laboratories and models of brain injury. At the clinical level, mild hypothermia has shown benefits in patients who have suffered cardiac arrest and in some pediatric populations suffering hypoxic brain insults. However, a review of the literature has demonstrated that in order to appreciate the maximum benefits of hypothermia, brain cooling needs to begin soon after the insult, maintained for relatively long period periods of time, and, in the case of ischemic stroke, should be applied in conjunction with the re-establishment of cerebral perfusion. Translating this to the clinical arena can be challenging, especially rapid cooling and the re-establishment of perfusion. The addition of a second neuroprotectant could potentially (1) enhance overall protection, (2) prolong the temporal therapeutic window for hypothermia, or (3) provide protection where hypothermic treatment is only transient. Combination therapies resulting in recanalization following ischemic stroke would improve the likelihood of a good outcome, as the experimental literature suggests more consistent neuroprotection against ischemia with reperfusion, than ischemia without. Since recombinant tissue plasiminogen activator (rt-PA) is the only FDA approved treatment for acute ischemic stroke, and acts to recanalize occluded vessels, it is an obvious initial strategy to combine with hypothermia. However, the effects of thrombolytics are also temperature dependent, and the risk of hemorrhage is significant. The experimental data nevertheless seem to favor a combinatorial approach. Thus, in order to apply hypothermia to a broader range of patients, combination strategies should be further investigated. PMID- 19216637 TI - Interaction of intracorporeal lithotripters with Proteus mirabilis inoculated inside artificial calcium and struvite stones. AB - BACKGROUND AND PURPOSE: Renal calculi may contain bacteria that can remain active inside the stone and produce bacteremia and/or endotoxemia after lithotripsy. Urinary tract infection associated with urinary stones represents high morbidity. The purpose of this research was to use novel artificial struvite stones inoculated with living bacteria and to study the effect of four different intracorporeal lithotripters on bacterial inactivation after in vitro lithotripsy. MATERIALS AND METHODS: Two types of artificial kidney stone models (calcium sulphate and mixed struvite-calcium sulphate) were manufactured and infected with Proteus mirabilis. Stones were fractured using either electrohydraulic, laser, ultrasonic, or pneumatic lithotripters. Bacterial viability was determined before and after the lithotripsy. RESULTS: Bacterial inactivation was not affected by the stone matrix; ie, calcium or struvite. The four tested lithotripters were almost equally efficient at reducing the viability of P mirabilis in both the low and the high energy setting. CONCLUSIONS: We were able to obtain novel artificial struvite stones infected with bacteria. Intracorporeal lithotripters are efficient at reducing the viability of P mirabilis in vitro. Tested stone materials play a minor role regarding inactivation. Whether the bactericidal effect reported is desirable or not is still to be answered, because the presence of endotoxin from cell lysis may increase the risk of urosepsis. PMID- 19216636 TI - Strain differences in response to traumatic brain injury in Long-Evans compared to Sprague-Dawley rats. AB - The selected strain of rodent used in experimental models of traumatic brain injury is typically dependent upon the experimental questions asked and the familiarity of the investigator with a specific rodent strain. This archival study compares the injury responsiveness and recovery profiles of two popular outbred strains, the Long-Evans (LE) and the Sprague-Dawley (SD), after brain injury induced by lateral fluid percussion injury (LFPI). General findings include a significantly longer duration of unconsciousness in LE rats, but similar durations of apnea. Both strains displayed the same level of initial FPI induced behavioral deficits, followed by a more rapid rate of functional recovery in SD rats. Cortical volume loss was not significantly different, but close inspection of the data suggests the possibility that LE rats may be more susceptible to damage in the hemisphere contralateral to the injury site than are SD rats. It is hoped that the information provided here encourages greater attention to the subtle differences and similarities between strains in future pre-clinical efficacy studies of traumatic brain injury. PMID- 19216638 TI - Transperitoneal laparoscopic repair of iatrogenic vesicovaginal fistulas: Heilbronn experience and review of the literature. AB - BACKGROUND AND PURPOSE: Vesicovaginal fistula (VVF) continues to be a distressing problem in urogynecology. Laparoscopy has become increasingly popular in the field of reconstructive urologic surgery. We describe our technique of laparoscopic VVF repair for iatrogenic VVFs, discuss our technique and outcomes, and review the related literature. PATIENTS AND METHODS: After hysterectomy for a benign condition, an iatrogenic VVF developed in three women (41, 48, and 34 years old, respectively) that persisted despite conservative therapy. Preoperative urethrocystoscopy demonstrated supratrigonal VVFs. Laparoscopic surgery was performed for the three patients after 3, 8, and 4 months of the previous abdominal operations. Patients were placed in a deflected supine position, and a transperitoneal five-port approach was used. The bladder was bivalved, and the fistula tract was excised. The fistulous tract was successfully repaired by closing the vagina and bladder separately. An interposing peritoneal flap was placed and fixed between the bladder and vagina. RESULTS: The patients recovered uneventfully and were discharged from the hospital after 4, 9, and 5 days, respectively. Retrograde cystography on postoperative day 10 revealed no contrast leakage via the vagina. The urethral catheters were removed on the same day. After 30, 17, and 14 months of follow up, respectively, the patients were voiding without any urine leakage. CONCLUSIONS: Laparoscopic repair can be applied effectively in the management of VVFs. The approach is safe and provides all advantages of minimally invasive surgery. Shorter hospital stay and shorter recovery time have a positive effect on the patients' well being. PMID- 19216640 TI - Controlled release of matrix metalloproteinase 1 with or without skeletal myoblasts transplantation improves cardiac function of rat hearts with chronic myocardial infarction. AB - Skeletal myoblast transplantation has been applied clinically for severe ischemic cardiomyopathy. Matrix metalloproteinase 1 (MMP-1) reduces fibrosis and prevents the progress of heart failure. We hypothesized that MMP-1 administration to the infarct area enhances the efficacy of skeletal myoblast transplantation. The controlled release of MMP-1 improved cardiac functions of rats with chronic myocardiac infarction with or without transplantation of skeletal myoblasts. Improvement in cardiac function and small fibrotic area inside the infarcted area were observed compared with those of myoblast transplantation. In conclusion, controlled release of MMP-1 was effective in cardioprotection in postmyocardial infarction although the combination with cell transplantation showed the similar effect. PMID- 19216641 TI - Improved cartilage regeneration utilizing mesenchymal stem cells in TGF-beta1 gene-activated scaffolds. AB - Recently, bone marrow-derived mesenchymal stem cells (MSCs) have been paid more attention for cartilage regeneration. This study evaluated the potential of using MSCs seeded in plasmid transforming growth factor beta1 (pTGF-beta1)-activated three-dimensional chitosan/gelatin scaffolds for improving cartilage repair in vivo. Significant cell proliferation and transforming growth factor beta1 protein expression were observed in vitro in pTGFbeta1-activated scaffolds. Transforming growth factor beta1-activated scaffolds showed high collagen type II and aggrecan expression and low collagen type I expression during in vitro cultivation. MSC based pTGF-beta1-activated scaffolds also exhibited cartilage histology with high secretion of collagen type II in vitro under the stimulation of pTGF-beta1. In rabbits with full-thickness cartilage defects, the implantation of MSC-based pTGF beta1-activated scaffolds not only significantly promoted chondrogenic differentiation of MSCs and hyalin-like cartilage matrix synthesis, but also remarkably improved the overall repair of rabbit cartilage defects and exhibited favorable tissue integrity at 10 weeks postsurgery. These results suggest that MSC-based localized pTGF-beta1-activated scaffolds have potential applications for in vivo cartilage repair. PMID- 19216642 TI - Buffered platelet-rich plasma enhances mesenchymal stem cell proliferation and chondrogenic differentiation. AB - The success of tissue engineering applications can potentially be dramatically improved with the addition of adjuncts that increase the proliferation and differentiation of progenitor or stem cells. Platelet-rich plasma (PRP) has recently emerged as a potential biologic tool to treat acute and chronic tendon disorders. The regenerative potential of PRP is based on the release of growth factors that occurs with platelet rupture. Its autologous nature gives it a significant advantage in tissue engineering applications. To test whether PRP may be useful specifically for cartilage regeneration, a cell culture experiment was devised in which mesenchymal stem cells (MSCs) were grown in control media or media enhanced with inactivated, buffered PRP. Proliferation 7 days after PRP treatment was increased: 1.041 versus 0.199 for the control media cells ( p<0.001). The messenger RNA (mRNA) level of the osteogenic marker RUNX2 was 52.84 versus 26.88 for the control group ( p<0.005). Likewise the mRNA level of the chondrogenic markers Sox-9 and aggrecan was 29.74 versus 2.29 for the control group ( p<0.001) and 21.04 versus 1.93 ( p<0.001), respectively. These results confirm that PRP enhances MSC proliferation and suggest that PRP causes chondrogenic differentiation of MSC in vitro. PMID- 19216643 TI - Three-dimensional loading model for periodontal ligament regeneration in vitro. AB - In this study we present a new three-dimensional (3D) model to study effects of mechanical loading on tendon/ligament formation in vitro. The model mimics a functional periodontal ligament (PDL), which anchors dental roots to the jaw bone and transfers the axial load of mastication to the jaw bone. A collagen gel containing human PDL fibroblasts was seeded in a PDL space between an artificial root and bone surface. The effects of 3-day loading on the fibroblasts were studied in vitro by axial and intermittent displacement of the root to which the gel was attached. Cell responses were recorded by measuring expression of three sets of genes: (i) cyclooxygenase 1 and 2 (COX-1, COX-2) producing prostaglandins (signaling molecules); (ii) Runx2, a transcription factor for the osteogenic lineage; and (iii) the extracellular matrix proteins osteopontin, dentin matrix protein 1, and collagen type I (COL1). Loading for 3 days resulted in magnitude dependent changes in the expression of COX-2 and COL1. A low loading magnitude significantly decreased COX-2 expression, an intermediate magnitude increased its expression, while a high magnitude increased COL1 expression. We concluded that the 3D loading model provides a useful, well-controlled method to examine ligament fibroblast responses to mechanical loading. The model may serve to explore the application of mechanical loading as an anabolic factor for ligament reconstruction. PMID- 19216644 TI - Modulation of the antifungal activity of new medicinal plant extracts active on Candida glabrata by the major transporters and regulators of the pleiotropic drug resistance network in Saccharomyces cerevisiae. AB - The increased incidence of drug-resistant fungal infections, a process in which active efflux plays an important role, calls for the development of new treatments. Candida albicans and Candida glabrata are the most frequent human fungal pathogens. The latter, in spite of its increased azole tolerance, is rarely used in medicinal plant screening. Several extracts inhibiting the growth of this pathogenic yeast are identified here. The ethyl acetate extract of the herb Dalea formosa of the American Southwest, not previously known to possess antifungal activity, proved most active against azole-sensitive and azole resistant isolates. The model yeast Saccharomyces cerevisiae, related to C. glabrata, was used to evaluate the influence of multidrug efflux on the antifungal activity of identified extracts and selected fractions from further purification steps, together with their ability to modulate ketoconazole resistance. The differential involvement of the major pleiotropic drug transporters of the ATP-binding cassette superfamily Pdr5p, Snq2p, and Yor1p as well as their transcriptional activators Pdr1p and Pdr3p in the detoxification of the antifungal constituents of several important medicinal plants is demonstrated. These include Artemisia annua and its widely used antimalarial component artemisinin. This approach revealed the concomitant presence of multidrug efflux pump substrates and modulators in the extract of A. annua and also allowed the identification of an extract not affected by the major pleiotropic drug-resistance genes. PMID- 19216645 TI - Antimicrobial resistance and virulence factors stx1, stx2, and eae in generic Escherichia coli isolates from calves in western Canadian cow-calf herds. AB - The objective of this study was to evaluate the association between antimicrobial resistance (AMR) in 106 fecal generic Escherichia coli isolates from calves in cow-calf herds, based on either phenotype or the presence of resistance genes, and the occurrence of virulence factors stx1, stx2, and eae. Three virulence genes and 23 AMR genes for six antimicrobial families were examined using DNA hybridization and PCR. Antimicrobial susceptibility of the isolates was tested using microbroth dilution (Sensititre, TREK Diagnostic Systems, Cleveland, OH) and the 2002 National Antimicrobial Monitoring System (NARMS) panel. The 106 isolates examined in this study were a stratified random subset from a larger study of AMR in cow-calf herds; 88.7% of the selected isolates were resistant to at least one antimicrobial, and 89.6% of the selected isolates were positive for at least one resistance gene. At least one virulence factor was identified in 48.1% (95%CI, 37.7-58.7) of the isolates. The most common virulence gene detected was stx2 followed by eae. Neither AMR measured phenotypically nor the presence of AMR genes were associated with the presence of above Shiga-toxigenic E. coli virulence factors in this population of healthy beef calves. PMID- 19216646 TI - Comparative antimicrobial susceptibility of Listeria monocytogenes, L. innocua, and L. welshimeri. AB - The current study compared antimicrobial susceptibility of Listeria innocua, L. welshimeri, and L. monocytogenes isolated from various sources. Antimicrobial susceptibility testing was performed using a microbroth procedure with Sensititre minimum inhibitory concentration plates containing 18 antimicrobials. Resistant isolates were analyzed for the presence of antimicrobial resistance genes using PCR. The majority of L. monocytogenes isolates were resistant to oxacillin (99%, 89/90) and ceftriaxone (72%, 65/90), while few isolates were resistant to clindamycin (21%, 19/90) and ciprofloxacin (2%, 2/90). When selected sources of L. monocytogenes are compared, resistance to ceftriaxone, clindamycin, and oxacillin ranged from 27% to 86%, 7% to 43%, and 96% to 100%, respectively. Resistance to ciprofloxacin (6%, 2/34), quinupristin/dalfopristin (7%, 1/14), and tetracycline (7%, 1/15) was observed with L. monocytogenes isolated from food, animal, and environmental sources, respectively. All L. welshimeri isolates (6/6) were resistant to streptomycin, quinupristin/dalfopristin, ciprofloxacin, rifampin, oxacillin, penicillin, and clindamycin, while most isolates (67%, 4/6) were resistant to trimethoprim/sulfamethoxazole. All L. innocua isolates (4/4) were resistant to oxacillin and penicillin, whereas 75% (3/4) of isolates were resistant to tetracycline, ceftriaxone, and clindamycin. Resistant isolates were negative for aadA, strA-B, sul I-II, penA, vat(A-E), vga(A-B), and vgb(A-B). However, tetM was detected among tetracycline-resistant isolates. L. welshimeri was resistant to more of the tested antimicrobials than the other two Listeria species tested, but resistance was not attributed to selected resistance genes. These data demonstrate the variability in resistance among Listeria species. However, the human pathogen L. monocytogenes appears to be the least resistant among the tested species. PMID- 19216647 TI - Antimicrobial resistance in Staphylococcus pseudintermedius in the Norwegian dog population. AB - The occurrence of antimicrobial resistance in Staphylococcus pseudintermedius from skin and ear infections in a representative sample of dogs unexposed to antimicrobial treatment before sampling was examined. The obtained isolates were further examined for genetic polymorphism and genetic background of resistance. A total of 59 isolates of S. pseudintermedius originating from 96 samples of 91 dogs in five different regions in Norway were included in this study. Susceptibility testing was performed using a broth dilution method. Resistant isolates were subjected to polymerase chain reaction for detection of resistance genes. All isolates were subjected to pulsed-field gel electrophoresis to examine the genetic polymorphism. In total, 19% of the isolates were susceptible to all antimicrobial agents included. Resistance to penicillin was most prevalent (70%), followed by resistance to fusidic acid (49%) and oxytetracycline (42%). Resistance to quinolones or cephalosporins was not observed. Resistance to penicillin, tetracycline, and erythromycin was mediated by the blaZ beta lactamase gene, the tetM gene, and the ermB gene, respectively. One of the fusidic acid-resistant isolates harbored a fusC gene, whereas the mechanisms involved in resistance in the other fusidic acid-resistant isolates remained unknown. Pulsed-field gel electrophoresis showed a high genetic polymorphism of S. pseudintermedius. This study indicates that the occurrence of antimicrobial resistance is common among S. pseudintermedius from dogs unexposed to antimicrobial treatment before sampling, and that there is a high genetic polymorphism among S. pseudintermedius. PMID- 19216648 TI - Carbonic anhydrases in meningiomas: association of endothelial carbonic anhydrase II with aggressive tumor features. AB - OBJECT: Carbonic anhydrase (CA) II and IX are enzymes involved in pH homeostasis and have been shown to be upregulated in several types of cancer. In this study, the authors evaluate the expression of CA II and IX in meningiomas and assess their relationship to patient age, tumor type and grade, tumor sex hormone receptor status, tumor cell proliferation, and tumor recurrence. METHODS: This study was conducted in consecutive patients who underwent meningioma surgeries at Tampere University Hospital between 1989 and 1999. The expression of CA II and IX was studied immunohistochemically using a tissue microarray technique and specific antibodies. RESULTS: Immunohistological staining with CA II and IX was assessed in 443 primary and 67 recurrent tumor specimens. Of these samples, 455 were benign (WHO Grade I), 49 atypical (Grade II), and 6 malignant (Grade III). Endothelial cells in 14.8% of the tumors stained positively for CA II. Tumor cells were positive for CA IX in 11.6% of the cases. Endothelial CA II expression correlated with increasing histological grade (p=0.002), and tumor proliferation rates were higher in CA II+ versus CA II- cases (p=0.002). Androgen receptor negative tumors were found to be CA II+ significantly more often than androgen receptor-positive tumors (p=0.001). No associations were found with the CA IX enzyme. CONCLUSIONS: Carbonic anhydrase II positivity in the endothelium was associated with cell proliferation and malignancy grade. These results suggest that CA II expression is associated with malignant progression of meningiomas and could thus be a target molecule for anticancer therapy. PMID- 19216649 TI - Relationship between the ophthalmic artery and the dural ring of the internal carotid artery. Clinical article. AB - OBJECT: The ophthalmic artery (OphA) usually arises from the intradural internal carotid artery (ICA), and the extradural origin has also been known. However, the interdural origin is extremely rare. The purpose of this paper was to clarify the origin of the OphA in patients with a paraclinoid aneurysm in the ICA based on intraoperative findings. METHODS: The authors retrospectively examined 156 patients who underwent direct surgical treatment for 166 paraclinoid aneurysms during a 17-year period. Based on intraoperative findings, 119 ophthalmic arteries were analyzed with respect to their origins. RESULTS: The OphA originated from the intradural ICA on 102 sides (85.7%), extradural on 9 (7.6%), and interdural on 8 (6.7%). Although the extradural origin might be recognized preoperatively, it was difficult to distinguish the interdural origin of the OphA from the intradural one. CONCLUSIONS: The incidence of the interdural origin was 6.7% and was not as rare as the authors expected. Neurosurgeons should know the possible existence of the interdural origin of the OphA to section the medial side of the dural ring. PMID- 19216650 TI - Characterization of functional outcome and quality of life following subarachnoid hemorrhage in patients treated with and without nicardipine prolonged-release implants. AB - OBJECT: The use of nicardipine prolonged-release implants (NPRIs) is associated with a significant improvement in the therapy of patients suffering from aneurysmal subarachnoid hemorrhage (aSAH) regarding the occurrence and severity of cerebral vasospasm, new infarcts, and functional outcome (FO). Because quality of life (QOL) measurements more reliably seem to describe the patient's true condition, the present study was conducted to assess FO and QOL 1 year after aneurysm rupture in patients with and without NPRIs. METHODS: From the initial series of 32 patients, 18 were assessed 1 year after aSAH (7 of the control and 11 of the NPRI group). The patients underwent neurological investigation, a structured interview followed by a measurement of QOL (Mini-Mental State Examination [MMSE]; 36-Item Short Form Health Survey [SF-36]; and the Hamilton Depression Rating Scale). There were no intergroup differences in the patient characteristics (that is, localization of aneurysm, initial Hunt and Hess grade, or age). RESULTS: In addition to the previously reported improvement of the National Institutes of Health Stroke Scale and modified Rankin Scale scores, the NPRI group's Karnofsky Performance Scale and the MMSE scores were markedly to significantly improved (p < 0.05 [Karnofsky Performance Scale] and p = 0.053 [MMSE]). In contrast, anxiety, oblivion, and mild symptoms of depression were equally present in both study groups (p = 0.607 [anxiety]; p = 0.732 [oblivion]; and p = 0.509 [Hamilton Depression Rating Scale]). Furthermore, no intergroup differences were observed in any of the SF-36 domains. The scores in the SF-36 domains of Role-Physical, Vitality, and Role-Emotional were significantly reduced in the NRPI group compared with those observed in an age-matched control population (p < 0.001 [Role-Physical]; p = 0.001 [vitality]; and p = 0.01 [Role Emotional]). Considering consequent costs, no difference was detectable regarding the duration of in- and outpatient rehabilitation (p = 0.135 and 0.171, respectively) or the Prolo score (p = 0.094). CONCLUSIONS: Despite FO improvement in terms of a lower incidence of cerebral vasospasm, new infarcts, morbidity in the treatment of aSAH in patients with NPRIs, a patient's QOL seems to be related to the severity of the aSAH itself. PMID- 19216651 TI - Electrocorticography-guided resection of temporal cavernoma: is electrocorticography warranted and does it alter the surgical approach? AB - OBJECT: Cavernous hemangiomas associated with epilepsy present an interesting surgical dilemma in terms of whether one should perform a pure lesionectomy or tailored resection, especially in the temporal lobe given the potential for cognitive damage. This decision is often guided by electrocorticography (ECoG), despite the lack of data regarding its value in cavernoma surgery. The purpose of the present study was several-fold: first, to determine the epilepsy outcome following resection of cavernomas in all brain regions; second, to evaluate the usefulness of ECoG in guiding surgical decision making; and third, to determine the optimum surgical approach for temporal lobe cavernomas. METHODS: The authors identified from their surgical database 173 patients who had undergone resection of cavernomas. One hundred two of these patients presented with epilepsy, and 61 harbored temporal lobe cavernomas. Preoperatively, all patients were initially evaluated by an epileptologist. The mean follow-up was 37 months. RESULTS: Regardless of the cavernoma location, surgery resulted in an excellent seizure control rate: Engel Class I outcome in 88% of patients at 2 years postoperatively. Of 61 patients with temporal lobe cavernomas, the mesial structures were involved in 35. Among the patients with temporal lobe cavernomas, those who underwent ECoG typically had a more extensive parenchymal resection rather than a lesionectomy (p < 0.0001). The use of ECoG in cases of temporal lobe cavernomas resulted in a superior seizure-free outcome: 79% (29 patients) versus 91% (23 patients) of patients at 6 months postresection, 77% (22 patients) versus 90% (20 patients) at 1 year, and 79% (14 patients) versus 83% (18 patients) at 2 years without ECoG versus with ECoG, respectively. CONCLUSIONS: The surgical removal of cavernomas most often leads to an excellent epilepsy outcome. In cases of temporal lobe cavernomas, the more extensive the ECoG-guided resection, the better the seizure outcome. In addition to upholding the concept of kindling, the data in this study support the use of ECoG in temporal lobe cavernoma surgery in patients presenting with epilepsy. PMID- 19216652 TI - Reversal of neurological deficit after chemotherapy in BCL-6-positive neurolymphomatosis. Case report. AB - Neurolymphomatosis, the infiltration of the peripheral nervous system (PNS) by malignant lymphatic cells, is a rare condition whose prognosis and treatment are not fully characterized. The authors report the case of a 69-year-old, previously healthy man who had a 1-month history of progressive pain in his right arm and associated weakness of several muscles of the right upper extremity when they first examined him. Initial MR imaging of the right brachial plexus showed no abnormalities, but over 3 months, symptoms gradually progressed to almost complete plegia of his right upper extremity. Subsequent MR imaging of his right brachial plexus showed an enhancing mass of the posterior cord of the plexus that encroached on the other cords. Positron emission tomography confirmed the presence of a hypermetabolic lesion in the right axillary region and also detected an asymptomatic hot spot in the gastric wall. Biopsy of the gastric lesion demonstrated a CD20+, diffuse large B-cell lymphoma that was immunohistochemically positive for BCL-6 and negative for p16. The patient underwent 6 cycles of dose-adjusted etoposide-vincristine-doxorubicin cyclophosphamide-prednisone (EPOCH) and rituximab, intermixed with 3 cycles of high-dose intravenous and intrathecal methotrexate, and followed by 6 monthly doses of rituximab for consolidation. Follow-up MR imaging and PET of the plexus showed complete radiological response after 3 months of treatment, as demonstrated by normalization of brachial plexus caliber, contrast enhancement, and metabolic activity. Twenty-eight months after symptom onset and 20 months after beginning therapy, the patient was disease-free, had recovered most upper extremity neurological function, and had only minimal remaining weakness of the right wrist and finger extension. PMID- 19216653 TI - Monitoring of brain interstitial total tau and beta amyloid proteins by microdialysis in patients with traumatic brain injury. AB - OBJECT: Damage to axons contributes to postinjury disabilities and is commonly observed following traumatic brain injury (TBI). Traumatic brain injury is an important environmental risk factor for the development of Alzheimer disease (AD). In the present feasibility study, the aim was to use intracerebral microdialysis catheters with a high molecular cutoff membrane (100 kD) to harvest interstitial total tau (T-tau) and amyloid beta 1-42 (Abeta42) proteins, which are important biomarkers for axonal injury and for AD, following moderate-to severe TBI. METHODS: Eight patients (5 men and 3 women) were included in the study; 5 of the patients had a focal/mixed TBI and 3 had a diffuse axonal injury (DAI). Following the bedside analysis of the routinely measured energy metabolic markers (that is, glucose, lactate/pyruvate ratio, glycerol, and glutamate), the remaining dialysate was pooled and two 12-hour samples per day were used to analyze T-tau and Abeta42 by enzyme-linked immunosorbent assay from Day 1 up to 8 days postinjury. RESULTS: The results show high levels of interstitial T-tau and Abeta42 postinjury. Patients with a predominantly focal lesion had higher interstitial T-tau levels than in the DAI group from Days 1 to 3 postinjury (p < 0.05). In contrast, patients with DAI had consistently higher Abeta42 levels when compared with patients with focal injury. CONCLUSIONS: These results suggest that monitoring of interstitial T-tau and Abeta42 by using microdialysis may be an important tool when evaluating the presence and role of axonal injury following TBI. PMID- 19216654 TI - A new way: alleviating postembolization syndrome following transcatheter arterial chemoembolization. AB - BACKGROUND: Currently, most therapies of postembolization syndrome following transcatheter arterial chemoembolization (TACE) aim directly at a single symptom, thus leading to limitations. OBJECTIVES: To seek for a systematic approach to prevent and treat the syndrome, we carried out this study to observe the effect of ginsenosides (GS) and dexamethasone (Dex) in alleviating the postembolization syndrome following TACE. METHODS: In the randomized, double-blinded and controlled trial, 120 patients with primary liver cancer were divided into 4 groups, with 30 patients in each group. The changes of clinical symptoms and laboratory tests before TACE and on 3 and 7 days after TACE were observed. RESULTS: The results indicated that Dex combined with GS not only markedly decreased the occurrence ratio and duration of such symptoms as nausea, vomiting, and fever, but also significantly reduced levels of total bilirubin, glutamic oxaloacetic transaminase, and glutamic-pyruvic transaminase (AST) and improved the Child-Pugh stage of liver function as compared with single use of GS or Dex. CONCLUSIONS: In conclusion, although single use of Dex or GS may improve some indices of adverse effects after TACE, the combination of Dex and GS can systematically prevent and treat the postembolization syndrome following TACE. PMID- 19216655 TI - Acupuncture for menopausal hot flashes: a qualitative study about patient experiences. AB - OBJECTIVE: The aim of this study was to describe any changes in health experienced by postmenopausal women after having acupuncture treatment for hot flashes. MATERIALS AND METHODS: Our sample was drawn from women participating in a randomized controlled trial (ACUFLASH) investigating the effect of acupuncture on menopausal hot flashes. One hundred and twenty-seven (127) women from the intervention group who had received a course of 10 acupuncture treatments were asked to make a written statement about any kind of change they had perceived that they considered could be related to the acupuncture treatment. Qualitative data were analyzed using systematic text condensation. RESULTS: Many women reported a substantial impact from the treatment with respect to a reduction in frequency and intensity of hot flashes both by night and by day. Changes related to improved sleep pattern were also reported, and a variety of different bodily and mental changes were described (i.e., feeling in a good mood, not so run down, and calmer). Several women were uncertain whether any changes had occurred. A few reported feeling worse. CONCLUSIONS: Our results describe a variety of health changes that may not be revealed by limited outcome measures in acupuncture studies. Further analysis of the relationship between such bodily experiences could lead to the development of hypotheses or models for how the acupuncture effect is mediated in complex bodily systems, and also contribute to development of outcome measures relevant for acupuncture studies. PMID- 19216656 TI - Probable hepatotoxicity related to Nerium oleander extract in a patient with metastatic synovial sarcoma of the knee. PMID- 19216657 TI - A review on the effects of aromatherapy for patients with depressive symptoms. AB - PURPOSE: We reviewed studies from 2000 to 2008 on using essential oils for patients with depression or depressive symptoms and examined their clinical effects. METHODS: The review was conducted among five electronic databases to identify all peer-reviewed journal papers that tested the effects of aromatherapy in the form of therapeutic massage for patients with depressive symptoms. RESULTS: The results were based on six studies examining the effects of aromatherapy on depressive symptoms in patients with depression and cancer. Some studies showed positive effects of this intervention among these three groups of patients. CONCLUSIONS: We recommend that aromatherapy could continue to be used as a complementary and alternative therapy for patients with depression and secondary depressive symptoms arising from various types of chronic medical conditions. More controlled studies with sound methodology should be conducted in the future to ascertain its clinical effects and the underlying psychobiologic mechanisms. PMID- 19216658 TI - 2005 Sasangin Diagnosis Questionnaire for Mongolian (05'SDQ-M): test of reliability. AB - OBJECTIVES: This study was proposed to evaluate the reliability by 2005 Sasangin Diagnosis Questionnaire for Mongolian (05' SDQ-M). DESIGN: Questions from the 05'SDQ were translated into Mongolian, and the 05'SDQ-M was constructed. The questionnaire was administered to 193 Mongolians and they were asked to complete the questionnaire again 2 weeks after the first administration. To evaluate the internal consistency, Cronbach's alpha coefficient was calculated. A reliability analysis for each individual question was conducted using the test-retest method to verify the independence and agreement between the questions. To verify independence, a chi(2) test with crosstabs analysis and p-values was used. The agreement verification was conducted by using the Cohen kappa coefficient. In addition, the constitutional variable was defined as the constitutional attribute of each question, and Pearson's correlation coefficient was confirmed after testing the scale. RESULTS AND CONCLUSIONS: After testing the internal consistency of the scale, the consistency was shown to range from 0.59 to 0.67, suggesting that the questions on the questionnaire were reliable. The test-retest method showed that Pearson's correlation coefficients for the results of the two tests ranged from 0.65 to 0.80. The x(2) test results indicated that one independent item demanded close attention. Among the other 123 questions, the agreement test revealed that 80 questions (64.5%) showed common agreement. PMID- 19216659 TI - A content analysis of the quantity and accuracy of dietary supplement information found in magazines with high adolescent readership. AB - OBJECTIVES: The objective of this study was to examine the quantity and accuracy of dietary supplement (DS) information through magazines with high adolescent readership. DESIGN: Eight (8) magazines (3 teen and 5 adult with high teen readership) were selected. A content analysis for DS was conducted on advertisements and editorials (i.e., articles, advice columns, and bulletins). Noted claims/cautions regarding DS were evaluated for accuracy using Medlineplus.gov and Naturaldatabase.com. Claims for dietary supplements with three or more types of ingredients and those in advertisements were not evaluated. Advertisements were evaluated with respect to size, referenced research, testimonials, and Dietary Supplement Health and Education Act of 1994 (DSHEA) warning visibility. RESULTS: Eighty-eight (88) issues from eight magazines yielded 238 DS references. Fifty (50) issues from five magazines contained no DS reference. Among teen magazines, seven DS references were found: five in the editorials and two in advertisements. In adult magazines, 231 DS references were found: 139 in editorials and 92 in advertisements. Of the 88 claims evaluated, 15% were accurate, 23% were inconclusive, 3% were inaccurate, 5% were partially accurate, and 55% were unsubstantiated (i.e., not listed in reference databases). Of the 94 DS evaluated in advertisements, 43% were full page or more, 79% did not have a DSHEA warning visible, 46% referred to research, and 32% used testimonials. CONCLUSIONS: Teen magazines contain few references to DS, none accurate. Adult magazines that have a high teen readership contain a substantial amount of DS information with questionable accuracy, raising concerns that this information may increase the chances of inappropriate DS use by adolescents, thereby increasing the potential for unexpected effects or possible harm. PMID- 19216660 TI - Comparison of effects of ginger, mefenamic acid, and ibuprofen on pain in women with primary dysmenorrhea. AB - OBJECTIVES: To compare the effects of ginger, mefenamic acid, and ibuprofen on pain in women with primary dysmenorrhea. METHODS: This was a double-blind comparative clinical trial conducted from September 2006 to February 2007. Participants were 150 students (18 years old and over) with primary dysmenorrhea from the dormitories of two medical universities who were alternately divided into three equal groups. Students in the ginger group took 250 mg capsules of ginger rhizome powder four times a day for three days from the start of their menstrual period. Members of the other groups received 250 mg mefenamic acid or 400 mg ibuprofen capsules, respectively, on the same protocol. A verbal multidimensional scoring system was used for assessing the severity of primary dysmenorrhea. Severity of disease, pain relief, and satisfaction with the treatment were compared between the groups after one menstruation. RESULTS: There were not significant differences between groups in baseline characteristics, p > 0.05. At the end of treatment, severity of dysmenorrhea decreased in all groups and no differences were found between the groups in severity of dysmenorrhea, pain relief, or satisfaction with the treatment, p > 0.05. No severe side effects occurred. CONCLUSION: Ginger was as effective as mefenamic acid and ibuprofen in relieving pain in women with primary dysmenorrhea. Further studies regarding the effects of ginger on other symptoms associated with dysmenorrhea and efficacy and safety of various doses and treatment durations of ginger are warranted. PMID- 19216661 TI - The effect of beta-glucan on recurrent aphthous stomatitis. PMID- 19216662 TI - Randomized controlled trials of acupuncture for neck pain: systematic review and meta-analysis. AB - OBJECTIVES: The objectives of this study were to assess the effectiveness and efficacy of acupuncture in the treatment of neck pain. DATA SOURCES: The following computerized databases were searched from their inception to January 2008: MEDLINE (PubMed), ALT HEALTH WATCH (EBSCO), CINAHL, and Cochrane Central. REVIEW METHODS: Systematic review and meta-analysis were conducted on randomized controlled trials of acupuncture for neck pain. Two (2) reviewers independently extracted data concerning study characteristics, methods, and outcomes, as well as performed quality assessment based on the adapted criteria of Jadad. RESULTS: Fourteen (14) studies were included in this review. Meta-analysis was performed only in the absence of statistically significant heterogeneity among studies that were selected for testing a specific clinical hypothesis. While only a single meta-analysis was done in previous reviews, this review performed nine meta analyses addressing different clinical issues. Seven out of nine meta-analyses yielded positive results. In particular, the meta-analysis based on the primary outcome of short-term pain reduction found that acupuncture was more effective than the control in the treatment of neck pain, with a pooled standardized mean difference (SMD) of -0.45 (95% confidence interval [CI], -0.69 to -0.22). Moreover, the meta-analysis with a pooled SMD of -0.53 (95% CI, -0.94 to -0.11) showed that acupuncture was significantly more effective than sham acupuncture for pain relief. However, there was limited evidence based on the qualitative analysis of the trial data to support the above conclusions. We provided a detailed analysis on the issue of heterogeneity of the studies involved in meta analysis and examined the consistencies and inconsistencies among the present review and two other reviews conducted previously. CONCLUSIONS: The quantitative meta-analysis conducted in this review confirmed the short-term effectiveness and efficacy of acupuncture in the treatment of neck pain. Further studies that address the long-term efficacy of acupuncture for neck pain are warranted. PMID- 19216663 TI - Death in the desert hits home. PMID- 19216664 TI - Preventive systemic antibiotics in colorectal surgery. AB - BACKGROUND: Elective colon surgery has the highest rate of surgical site infection (SSI) of any elective procedure in surgery. The evolution of strategies for the prevention of SSI has had the correct use of preventive antibiotics as a crucial component. METHODS: Review of the pertinent English-language literature. RESULTS: Correct use of prophylactic antibiotics in elective colon surgery requires that the drug have activity against colonic aerobic and anaerobic bacteria, be administered immediately before the surgical incision, and not be continued into the postoperative period. All of the documented and many of the theoretically feasible antibiotic choices are discussed in terms of their advantages for this indication. CONCLUSIONS: The best outcomes in the prevention of SSI in elective colon surgery are achieved when appropriate systemic antibiotics are employed in conjunction with the appropriate use of oral antibiotic bowel preparation. PMID- 19216665 TI - Controversies in appendicitis. AB - BACKGROUND: Appendectomy for appendicitis is a common operation that is performed by most general surgeons. Despite the large number of appendectomies, numerous management controversies continue. METHODS: Review of the pertinent English language literature. RESULTS: Controversy persists regarding the use of imaging studies for diagnosis, laparoscopic vs. open surgical methods, nonoperative management of uncomplicated appendicitis, delayed management of the patient with phlegmon from severe appendicitis, the necessity for interval appendectomy, and the merits of deferral of off-hour appendectomy until the following morning. This review identifies the published reports that support the pros and cons of each controversy. CONCLUSIONS: Class I data generally are lacking to support the resolution of these controversies. Additional well-controlled clinical trials are necessary to obtain definitive answers. PMID- 19216666 TI - Necrotizing soft tissue infection. AB - BACKGROUND: Necrotizing soft tissue infections (NSTIs) continue to pose major problems in diagnosis and management. METHODS: Review of the pertinent English language literature. RESULTS: A high index of suspicion and good clinical judgment should be used in conjunction with laboratory and imaging studies in appropriate cases to establish the diagnosis as rapidly as possible. Successful treatment requires early, aggressive surgical debridement of all necrotic tissue, appropriate systemic antibiotic therapy, and supportive care to maintain oxygenation and tissue perfusion. CONCLUSIONS: The mortality rate of necrotizing soft tissue infection remains approximately 25% despite optimal care. Delayed definitive debridement remains the single most important risk factor for death. PMID- 19216667 TI - Mechanical bowel preparation or not for elective colorectal surgery. AB - BACKGROUND: Mechanical bowel preparation has been a mainstay in the management of patients undergoing elective colorectal surgery, but in recent years, this practice has been challenged; and a number of clinical reports have demonstrated no benefit, as judged by reduction of surgical site infections. This paper presents the advantages and disadvantages of mechanical bowel preparation. METHODS: Review of the pertinent English language literature. RESULTS: Some published studies support omission of mechanical preparation of the colon prior to elective colo-rectal surgery. However, the evidence is not definitive, in that the most rigorous study performed to date, a large, randomized trial, is non confirmatory. CONCLUSIONS: Additional stratified studies of different mechanical bowel preparation methods, compared with no preparation, are required to address whether the practice should be abandoned. PMID- 19216668 TI - Vancomycin-resistant enterococcus: risk factors, surveillance, infections, and treatment. AB - BACKGROUND: The use of vancomycin has continued to expand because of the increasing number of patients infected or colonized with methicillin-resistant Staphylococcus aureus, causing an increase in the prevalence of vancomycin resistant Enterococcus (VRE). METHODS: Review of the pertinent English language literature. RESULTS: Vancomycin-resistant Enterococcus spp. are being identified more often in nosocomial infections of surgical patients. The biology of resistance, modes of transmission, patient risk factors, and current treatment strategies are discussed. CONCLUSIONS: The reservoir of resistance in enterococci looms as a major threat for genetic transfer and the emergence of increasing numbers of vancomycin-resistant S. aureus. PMID- 19216669 TI - Severe acute pancreatitis: a review. AB - BACKGROUND: Severe acute pancreatitis continues to be a difficult problem in clinical management. This paper provides a contemporary definition of the condition and explores the controversial issues that surround its diagnosis and management. METHODS: Review of pertinent English language literature. RESULTS: The use of various imaging techniques is discussed, with particular emphasis on the assessment of pancreatic necrosis and the evaluation of choledocholithiasis as a cause of the pancreatitis. Prophylactic antibiotics generally are discouraged and early and aggressive nutritional support is advocated. Delayed surgical intervention is recommended to avoid the severe consequences associated with prematurely early attempts at resection of the infarcted pancreas and adjacent retroperitoneal fat. CONCLUSIONS: Better quality evidence, especially regarding the utility or lack thereof of antibiotic prophylaxis, is beginning to inform optimal management of patients with severe acute pancreatitis. PMID- 19216673 TI - Disease management grows up. PMID- 19216670 TI - Surgical site infections and the surgical care improvement project (SCIP): evolution of national quality measures. AB - BACKGROUND: Surgical site infection (SSI) continues to be a common if often times preventable infection. Considerable patient morbidity and economic cost are the consequences. METHODS: Review of the pertinent English-language literature. RESULTS: The National Surgical Infection Prevention (SIP) Project was an initiative sponsored jointly by the Centers for Medicare and Medicaid Services and the U.S. Centers for Disease Control and Prevention to decrease the incidence of SSI in major surgical procedures. Three performance measures were developed to improve the timing of antibiotic administration and the selection of the drug and to reduce needless postoperative administration of antibiotics. The success of SIP led to the Surgical Care Improvement Project (SCIP), which is expanding performance measures in the area of prevention of SSI, and also is undertaking preventive measures against postoperative venous thromboembolism and cardiac events. Federal legislation has required hospitals to report rates of compliance with many of these process measures. CONCLUSIONS: Surgeons must use all documented strategies to prevent SSIs. PMID- 19216674 TI - Using qualitative and quantitative methods to evaluate small-scale disease management pilot programs. AB - Interest in disease management programs continues to grow as managed care plans, the federal and state governments, and other organizations consider such efforts as a means to improve health care quality and reduce costs. These efforts vary in size, scope, and target population. While large-scale programs provide the means to measure impacts, evaluation of smaller interventions remains valuable as they often represent the early planning stages of larger initiatives. This paper describes a multi-method approach for evaluating small interventions that sought to improve the quality of care for Medicaid beneficiaries with multiple chronic conditions. Our approach relied on quantitative and qualitative methods to develop a complete understanding of each intervention. Quantitative data in the form of both process measures, such as case manager contacts, and outcome measures, such as hospital use, were reported and analyzed. Qualitative information was collected through interviews and the development of logic models to document the flow of intervention activities and how they were intended to affect outcomes. The logic models helped us to understand the underlying reasons for the success or lack thereof of each intervention. The analysis provides useful information on several fronts. First, qualitative data provided valuable information about implementation. Second, process measures helped determine whether implementation occurred as anticipated. Third, outcome measures indicated the potential for favorable results later, possibly suggesting further study. Finally, the evaluation of qualitative and quantitative data in combination helped us assess the potential promise of each intervention and identify common themes and challenges across all interventions. PMID- 19216675 TI - "Dial-an-ROI?" changing basic variables impacts cost trends in single-population pre-post ("DMAA type") savings analysis. AB - Disease management (DM) programs claim to achieve cost savings by reducing clinical adverse events. While measuring changes in adverse events is straightforward, plausibly demonstrating savings has been contentious, especially absent an external comparison population. In this situation, a single-population methodology is often used, in which the cost trend for those with no program conditions ("non-chronics"--NC) forms the expected trend for those who have at least 1 program condition ("chronics"--C). The methodology's fundamental assumption is that--absent DM--C and NC trends would be identical (or bear a constant relationship over time). We assessed this assumption by altering the values of key variables used to identify C and NC, and to calculate trend. We compared C and NC baseline trends for a 23-condition telephonic DM multiemployer program representing nearly 300,000 members. Trends were calculated for 16 combinations of values for 4 key variables: identification look-back frame (12 vs. 24 months); identification threshold (high vs. lower specificity); claims runout (3 vs. 6 months); and minimum required insurance eligibility (any 6 months vs. 12 months continuous eligibility in the measurement year). Identification was performed by annual qualification. Changes in values for the 4 key variables markedly impacted baseline C and NC trends. C trends varied between 10.1% and 13.1%; NC trends between 5.2% and 12.8%. C-NC trend differences ranged between 1.9% and +7.0%. The combination of 24 months identification look-back, high identification threshold, 6 months runout, and any-6-months eligibility gave the most convergent C and NC trends (10.4% and 10.7%). Seemingly minor changes in key variables impact C and NC trends in single-population pre-post DM savings methodologies. When a suitable comparison population is not available, at least 1 year of baseline C and NC trends should be reported-as recommended by the DMAA- and values of key variables used should be specified. Plausibility metrics (eg, hospitalizations) should be reported. PMID- 19216676 TI - Measuring medication adherence. AB - Several studies have shown that poor adherence to pharmaceutical treatment regimens increases health condition severity, mortality rates, and overall health care costs. The dimension of medication adherence in health management outcomes has been deficient of a comprehensive yet practical standardized measurement methodology. DMAA: The Care Continuum Alliance, along with its industry partners, noted this absence and assembled a clinical workgroup to undertake an appropriate standardization effort. The medication adherence clinical workgroup within DMAA's outcomes measurement and methodology steering committee convened in February 2008 to review literature, incorporate subject matter expertise, gain consensus on measurement techniques, and discuss known limitations. This workgroup has successfully completed a standardized methodology to measure medication adherence rates via medication possession ratio (MPR). The guidelines for this method were approved for inclusion in Volume III of DMAA's Outcomes Guidelines Report. Accordant Health Services, as a member of this workgroup, volunteered to test the recommended specification and measure. These tests would encompass not only measurement calculations but also feasibility considerations. The measurement period for these tests was the 2007 calendar year. Results showed that condition specific MPRs for a health plan population ranged between 78.9% and 83.6%. The same methods applied to an employer group population revealed a condition specific MPR range of 75.4% to 76.5%. Based on 2007 book of business data for CVS Caremark's Adherence to Drug Therapy program, those results are within the 95% confidence interval for employer group and health plan populations. PMID- 19216677 TI - The relationship between chronic and non-chronic trends. AB - Cost trend plays a crucial role in evaluating the outcome of disease management (DM) programs, which typically attempt to manage members who have any of 5 major chronic conditions (the chronic population) through targeted intervention. A widely adopted methodology for evaluating DM outcomes is commonly referred to as the "pre-post" method, also known as the adjusted historical control method. One drawback of the pre-post approach is the need for a valid trend adjustment so that program year and baseline year costs can be compared to determine the impact of the DM program. The trend adjustment plays a crucial role in the pre-post methodology because the estimated cost savings is very sensitive to the variation in the trend estimate. DMAA: The Care Continuum Alliance recommends that the concurrent trend for members who do not have any of the 5 chronic conditions (the non-chronic population) be used to estimate the chronic trend. One major assumption in using this method is that there is a relatively stable relationship between the chronic trend and the non-chronic trend for the same population in the absence of the DM program. In this analysis, the relationship between the two trends is studied based on empirical data. Some key factors that may impact the relationship between the chronic and non-chronic trends are also examined. Medical and pharmacy claims from a large commercial client over a 4-year period were collected for the analysis. This client did not have any robust DM program in place during the 4-year period. The major findings from this study include: (1) The method used to identify the chronic population for calculating the trend has a significant impact on the relationship between the chronic and non-chronic trends. With the Annual Qualification method, in which the chronic members are identified using the claims in a specific time period regardless of whether they have been identified previously or thereafter, the relationship between the non chronic and chronic trends appears stable over time. With the Prospective method, the non-chronic trend does not serve as a good estimate of the chronic trend. (2) It is important to use risk adjustment when calculating trends to account for risk changes within the population from year to year. The chronic and non-chronic trends converge further when risk adjustment is applied. Based on the empirical evidence from the data used in this study, the concurrent non-chronic trend could be used to help estimate the chronic trend if the proper member selection method (Annual Qualification) is used to select the chronic population and proper steps are taken to adjust the different risk levels from year to year. One should not conclude, however, that such a stable relationship between chronic and non chronic trend has been universally validated based on this single study. This study highlights the importance of carefully validating trend relationships from a particular population before assuming a stable trend relationship in DM program outcome evaluation. PMID- 19216678 TI - Challenges and solutions in the evaluation of a low back pain disease management program. AB - This paper examines a novel approach to evaluating a nurse-run telephonic low back pain (LBP) disease management (DM) program offered by a fully-insured commercial health plan population with approximately 150,000 members located in the northeastern United States. Members with at least 6 member months of eligibility were identified to have LBP using administrative claims and eligibility data. The LBP program relies on telephonic nurse management augmented by printed materials, adapting and advocating HEDIS imaging guidelines, and drug utilization review. Outcomes of this LBP DM program were assessed using a pre post population-based approach as recommended in the DMAA Outcomes Guidelines Report. Baseline year and program year populations were segmented into 5 LBP clinical categories and each was weight adjusted using population size. LBP related medical service utilization and pharmacy utilization also were evaluated. Individuals under active LBP management exhibited a decrease in LBP-related imaging and surgeries. Overall analgesic use also decreased. These data suggest the LBP DM program is associated with a decrease in imaging, surgeries, and analgesic use. The magnitude of the clinically segmented weight-adjusted return on investment (ROI) was lower than the unadjusted ROI, but was directionally positive indicating program impact. This adjustment is necessary in order to gain insight into and consistency of the results of a comprehensive LBP DM program evaluation. PMID- 19216679 TI - How to measure the outcomes of chronic disease management. AB - The fastest-growing methodology for disease management outcomes measurement is valid, transparent, easy to apply, and freely available in the public domain and this article. It measures the actual goal of disease management, which is to reduce the rate of adverse events associated with the disease(s) being managed. Changes in this rate can be translated into a return on investment using some explicit assumptions about comorbidities and episode costs. Outcomes measured in this way show that in the health plan community as a whole, disease management in the broadest sense is working, as measured by the relative stability in the rate of adverse medical events closely associated with common chronic disease during this decade of increasing prevalence of most of the common chronic conditions. PMID- 19216681 TI - Immune responses to herpesviral vectors. PMID- 19216682 TI - Genetic diversity of the low-level vancomycin resistance gene vanC-2/vanC-3 and identification of a novel vanC subtype (vanC-4) in Enterococcus casseliflavus. AB - An intrinsic low-level vancomycin resistance (VanC phenotype) in Enterococcus casseliflavus is conferred by either of two subtypes of vanC genes, that is, vanC 2 or vanC-3, which are genetically closely related. To know genetic diversity of vanC-2/C-3 genes among E. casseliflavus, nucleotide sequences of vanC-2/C-3 and other genetic components in vanC gene cluster (vanXYc, vanTc, vanRc, and vanSc) were analyzed for nine clinical isolates and four standard strains that showed low-level vancomycin resistance. While the vanC-2/C-3 gene sequences showed 93 100% identities among the strains examined, two genetic groups were discriminated by phylogenetic analysis: one closely related to the previously reported vanC-2 or vanC-3 genes (vanC-2/C-3 genotype) with 98-100% identity, and the other distinct from the vanC-2/C-3 genotype (93-95% identity). The latter group found in three clinical isolates was considered as a new subtype of vanC and tentatively designated as vanC-4. Between strains with the vanC-2/C-3 genotype and those with vanC-4, vanXYc genes were also genetically discriminated with 92 93% identity. Similar sequence diversity was observed for vanTc, vanRc, and vanSc (88-93% identity). Clonal relatedness among the E. casseliflavus strains was investigated by phylogenetic analysis of atpA gene. While among E. casseliflavus strains with vanC-2/C-3 genotype, extremely high sequence identities of atpA were found (98.7% or higher), these strains showed slightly lower identity to those with vanC-4 (94-96%). These two groups of E. casseliflavus strains were also discriminated by genotyping with arbitrarily primed PCR. These findings indicated that among E. casseliflavus there are at least two genetic lineages with the distinct vanC genes, that is, a single subtype including previously known vanC 2/C-3, and a novel subtype vanC-4. PMID- 19216683 TI - Atmosphere and water loss from early Mars under extreme solar wind and extreme ultraviolet conditions. AB - The upper limits of the ion pickup and cold ion outflow loss rates from the early martian atmosphere shortly after the Sun arrived at the Zero-Age-Main-Sequence (ZAMS) were investigated. We applied a comprehensive 3-D multi-species magnetohydrodynamic (MHD) model to an early martian CO(2)-rich atmosphere, which was assumed to have been exposed to a solar XUV [X-ray and extreme ultraviolet (EUV)] flux that was 100 times higher than today and a solar wind that was about 300 times denser. We also assumed the late onset of a planetary magnetic dynamo, so that Mars had no strong intrinsic magnetic field at that early period. We found that, due to such extreme solar wind-atmosphere interaction, a strong magnetic field of about approximately 4000 nT was induced in the entire dayside ionosphere, which could efficiently protect the upper atmosphere from sputtering loss. A planetary obstacle ( approximately ionopause) was formed at an altitude of about 1000 km above the surface due to the drag force and the mass loading by newly created ions in the highly extended upper atmosphere. We obtained an O(+) loss rate by the ion pickup process, which takes place above the ionopause, of about 1.5 x 10(28) ions/s during the first < or =150 million years, which is about 10(4) times greater than today and corresponds to a water loss equivalent to a global martian ocean with a depth of approximately 8 m. Consequently, even if the magnetic protection due to the expected early martian magnetic dynamo is neglected, ion pickup and sputtering were most likely not the dominant loss processes for the planet's initial atmosphere and water inventory. However, it appears that the cold ion outflow into the martian tail, due to the transfer of momentum from the solar wind to the ionospheric plasma, could have removed a global ocean with a depth of 10-70 m during the first < or =150 million years after the Sun arrived at the ZAMS. PMID- 19216684 TI - Continuous glucose monitoring in non-insulin-using individuals with type 2 diabetes: acceptability, feasibility, and teaching opportunities. AB - BACKGROUND: Continuous glucose monitoring (CGM) has the potential to provide useful data for behavioral interventions targeting non-insulin-using, sedentary individuals with type 2 diabetes mellitus (T2DM). The aims of this study were to describe CGM in terms of (1) feasibility and acceptability and (2) dietary- and exercise-teaching events. METHODS: Cross-sectional data were analyzed from 27 non insulin-using adults with T2DM who wore CGM for 72 h as part of a larger study on using CGM for exercise counseling in this population. Feasibility data included accuracy of entering daily self-monitored blood glucose (SMBG) readings and events (e.g., meals, exercise), sensor failures, alarms, optimal accuracy of glucose data, and download failures. Acceptability data included CGM satisfaction and wearing difficulties. Dietary- and exercise-teaching events were identified from CGM and activity monitor data. RESULTS: CGM graphs showed 141 dietary- and 71 exercise-teaching events. About half the participants (52%) reported difficulty remembering to enter events into CGM monitors, but most (82%) kept an accurate paper log of events. Insufficient SMBG entries resulted in 32 CGM graphs with "use clinical judgment" warnings. Eighty-three percent of missed SMBG entries were from 18 participants 55-77 years old. Missing correlation coefficients resulted from glucose concentrations varying <100 mg/dL. A majority of participants (n = 19) were willing to wear CGM again despite reporting minor discomfort at sensor site and with wearing the monitor. CONCLUSIONS: CGM data provided several teaching opportunities in non-insulin-using adults with T2DM. Overall, CGM was acceptable and feasible. Some identified problems may be eliminated by newer technology. PMID- 19216685 TI - An automated telemedicine system improves patient-reported well-being. AB - BACKGROUND: Control of serum glucose levels is essential for the reduction of complications of diabetes. Telemedicine is one strategy through which serum glucose control can be improved. METHODS: A total of 35 adult, insulin-treated patients with diabetes (type 1 and type 2) were enrolled in the present study (63.0 +/- 10 years of age, 63% female) and randomized to telemedicine monitoring (including cordless, remote glucose monitor, and transmitter, n = 17), or conventional follow-up (n = 18). Metabolic parameters were evaluated, and a quality of life questionnaire was administered both pre- and post-treatment. RESULTS: Groups were similar at baseline in terms of demographic, quality of life, and metabolic parameters. Significant differences in post-treatment metabolic parameters were not observed, although serum glucose was marginally elevated in the control group compared to the telemedicine group (214 +/- 65 mg/dL vs. 171 +/- 77 mg/dL, P = 0.09). On the other hand, being clinically symptom-free (71% vs. 11%, P = 0.003), having no hypoglycemic events (82% vs. 17%, P = 0.0001), and having no hyperglycemic events (65% vs. 17%, P = 0.004) were all significantly more frequently reported in the telemedicine group compared to the control group. Compared to the control group, the telemedicine group reported experiencing significantly less anxiety, treatment difficulty, depression, disease-associated life complications, and feelings of impotence or ineptitude and significantly greater improvement in personal control over glucose, weight, and overall diabetes. CONCLUSIONS: Though post-treatment metabolic differences were not observed between treatment groups, the telemedicine group reported significantly greater post-treatment experiences of improved quality of life and sense of control over the disease. Thus patient satisfaction can be enhanced through the use of telemedicine. PMID- 19216686 TI - Detection of hypoglycemia with continuous interstitial and traditional blood glucose monitoring using the FreeStyle Navigator Continuous Glucose Monitoring System. AB - BACKGROUND: The objective of the analysis was to compare detection of hypoglycemic episodes (glucose <70 mg/dL lasting >15 min) with the FreeStyle Navigator Continuous Glucose Monitoring System (FSN-CGM) (Abbott Diabetes Care, Alameda, CA) alarms to detection with traditional finger stick testing at an average frequency of eight tests per day. METHODS: The performance of FSN-CGM alarms was evaluated in a clinic setting using 58 subjects with type 1 diabetes mellitus (T1DM) monitoring interstitial glucose concentration over a 5-day period compared to reference YSI measurements (instrument manufactured by YSI, Yellow Springs, OH) at 15-min intervals. Finger stick glucose testing was evaluated in the home environment with 91 subjects with TIDM monitoring with the blood glucose meter integrated into the FreeStyle Navigator (FSN-BG) over a 20-day period. The reference was FSN-CGM with results masked from the subjects. Blood glucose values <=85 mg/dL were considered the optimal treatment level to avoid or reverse hypoglycemia. RESULTS: With a threshold alarm setting of 85 mg/dL, 90.6% of hypoglycemic episodes were detected within +/- 30 min by FSN-CGM in the clinic study. When the alarm was activated, YSI glucose was <= 85 mg/dL 77.2% of the time. In the home environment, the average FSN-BG testing frequency was 7.9 tests per day. Hypoglycemia was verified within +/- 30 min by FSN-BG measurements <= 85 mg/dL at a rate of 27.5%. CONCLUSIONS: Even with a high rate of FSN-BG testing, hypoglycemia detected by FSN-CGM was verified by patients with T1DM very infrequently. A high rate of hypoglycemia detection with a moderate rate of unnecessary alarms can be attained using FSN-CGM. PMID- 19216687 TI - Graphical and numerical evaluation of continuous glucose sensing time lag. AB - AIM: This study introduces a new method for graphical and numerical evaluation of time lags typically associated with subcutaneous glucose sensing, based on Poincare-type plot and a maximum statistical agreement criterion. METHODS: The proposed method is illustrated by retrospective analysis of 56 continuous glucose monitor (CGM) time series collected by the FreeStyle Navigator (Abbott Diabetes Care, Alameda, CA) from 28 patients with type 1 diabetes mellitus, each wearing simultaneously two sensors (on arm and abdomen) and parallel reference blood glucose (BG) collected with a reference YSI (Yellow Springs, OH) analyzer every 15 min. The average duration of a time series was 111 h; there were approximately 10,000 sensor-reference data pairs. RESULTS: When sliding in time CGM readings versus BG, the point of minimal spread of a Poincare-type plot marks visually the time of CGM delay. The same point is numerically estimated by minimizing the distance between BG and CGM readings. The average observed time lag between reference BG and CGM was 12.5 min. Stratified by BG rate of change, the time lag was longer (16.8 min) when BG was falling, compared to steady or rising BG (11.7 min and 9.9 min, respectively) (P < 0.005). The time lags at the two sensor locations were not significantly different: 12.4 min on the arm, 12.6 min on the abdomen. CONCLUSIONS: In this data set, substantial blood-to-sensor time delays were observed, possibly because of both blood-to-interstitial glucose transport and instrumental delay. Analysis of BG-CGM co-dynamics that is free from mathematical approximation of glucose fluctuations resulted in convenient visualization and numerical estimation of these delays. PMID- 19216688 TI - The creation and impact evaluation of "Your guide to diet and diabetes," an interactive web-based diabetes tutorial. AB - BACKGROUND: Millions of Americans turn to the Internet for health information. A need for a comprehensive website that provides information on diet and diabetes management was identified. Therefore, the goal of this study was to develop an easy-to-use website that provided accurate information about diet and diabetes management in an interactive format that was widely accessible. METHODS: The "Your Guide to Diet and Diabetes" website was developed through issue identification, expert panel review, and face validity phase for content and knowledge/skill assessment surveys. During the first 2 weeks of the website's availability, impact data were collected from website visitors. Responses to demographic questions, knowledge, and skill questions were collected in a database. RESULTS: Of the 406 individuals who completed a demographic question, a maximum of 104 respondents chose to answer one or more of the interactive knowledge or skills questions. CONCLUSIONS: "Your Guide to Diet and Diabetes" filled a need for an easy-to-use website that addressed the issues of diet and diabetes. Based on the number of visits to the login page (n = 6,124) and number of individuals who answered demographic questions, the site was well received. While a smaller group of participants elected to complete the interactive knowledge and skills questions, responses to these questions demonstrated that participants obtained knowledge from the website. In particular, participants increased their knowledge of food sources and appropriate serving sizes of carbohydrate. The "making a plate" skill activity was used with much greater frequency than the more conceptually complex carbohydrate counting skill activity. PMID- 19216689 TI - Factors associated with adherence to continuous subcutaneous insulin infusion in pediatric diabetes. AB - BACKGROUND/AIMS: Continuous subcutaneous insulin infusion (CSII) is a safe and effective alternative to insulin injections in pediatric type 1 diabetes mellitus. CSII can be associated with an increased risk of hypoglycemia and diabetic ketoacidosis (DKA) in some patients. In our Center, patients/guardians are screened for proficiency in diabetes management skills as a prerequisite to initiation of CSII. We reviewed the clinical data from our patients to assess the predictors associated with nonadherence to CSII therapy. METHODS: We retrospectively collected clinical data on all our CSII initiations between July 1999 to June 2003, including: body mass index, hemoglobin A1c (HbA1c), total daily dose, bolus to basal insulin ratio, hypoglycemic episodes (blood glucose <60 mg/dL/week), mean fasting self-monitored blood glucose (SMBG), severity of lipohypertrophy, DKA, and pubertal status. RESULTS: Forty-six patients 9.90 +/- 3.4 years old (28 girls and 18 boys) started CSII in the 4-year period. While 39 patients (85%) 9.8 +/- 3.5 years old currently remain on CSII, seven patients (15%) 11.2 +/- 0.9 years old discontinued CSII. Fifteen patients (32.6%) were prepubertal at CSII initiation, and none discontinued CSII in this cohort, whereas seven of 31 (22.6%) pubertal patients discontinued CSII. The patients who continued CSII were similar to the CSII-discontinued cohort at baseline. At 12 months, rising HbA1c was the only predictor of future nonadherence to CSII. At 24 months, the discontinuation group had higher mean fasting SMBG levels and severe lipohypertrophy (P < 0.05). None of the prepubertal patients discontinued CSII, while all seven patients (100%) in the CSII-discontinued group were pubertal (P < 0.001). CONCLUSIONS: Extensive screening by a multidisciplinary diabetes team prior to initiation of CSII regimen results in relatively lower discontinuation rates and a higher chance of maintaining optimal glycemic control (HbA1C < 8%) compared to previous studies. PMID- 19216690 TI - Decreased recurrence rate in the laparoscopic herniorraphy in children: comparison between two techniques. AB - AIM: The laparoscopic herniorraphy in children is still associated to a high recurrence rate. The aim of this study was to assess whether the addition of the lateral incision of the sac to the sole suture of the inner inguinal ring could reduce the recurrence rate. MATERIALS AND METHODS: A retrospective review was performed of the collected data of 248 laparoscopic inguinal hernia repairs in 224 children (175 males, 49 females) between 8 months and 11 years of age (mean age, 5 years; median, 4) in our institution from January 2004 to December 2007. The hernia was unilateral in 204 patients (133 on the right side, 71 on the left) and bilateral in 20 patients. A 5-mm umbilical camera port for a 0-degree laparoscopic optics and two operative 2- or 3-mm reusable trocars inserted in the lower right and left quadrants of the abdominal wall were utilized. In a group of 123 patients, the inner inguinal ring was closed, adopting a W-shaped suture (inguinal ring suture; IRS). In the other group of 101 patients, a lateral incision of the sac of 1-2 cm was carried out before the W-shaped suture of the inner inguinal ring (inguinal ring incision suture; IRIS). RESULTS: At a mean follow-up of 24 months (range, 6-36), 5 of 133 (3.76%) hernias recurred between 6 and 12 months after surgery in the IRS group. In the IRIS group, none of the patients presented with recurrence. The rate of recurrences in the two groups was compared and analyzed with the x2 test. The resulting difference was statistically significant (P < 0.05). CONCLUSION: In our experience, the incision of the peritoneum lateral to the internal inguinal ring and the W-shaped suture, compared to the sole W-shaped suture, is safe and effective in preventing hernia recurrence. PMID- 19216691 TI - Robot-assisted thoracoscopic resection of intralobar sequestration. AB - In this paper, we report the first case of a robot-assisted thoracoscopic resection of intralobar sequestration. By virtue of greater dexterity and three dimensional visualization, the da Vinci robot enables a safer, more precise dissection of sequestered tissue in the face of chronic inflammatory adhesions than conventional video-assisted thoracoscopic surgery does. Thus, in expert hands, such robotic technology is likely to result in less bleeding complications and less conversions to open surgery in cases of sequestration. PMID- 19216693 TI - A case of concomitant pneumothorax and pneumoperitoneum. AB - Pneumothorax associated with pneumoperitoneum is rare. There are scattered case reports of this syndrome in the pediatric literature but very few reports in the adult literature. Several possible causes exist. Maurer at al. suggested that it results from air entering the pleural space through a diaphragmatic defect. In this paper, we report a patient with a perforated diverticulum and a concomitant pneumothorax. PMID- 19216692 TI - Laparoscopic revision of failed fundoplication and hiatal herniorraphy. AB - OBJECTIVE: The aim of this study was to evaluate the mechanisms of failure after laparoscopic fundoplication and the results of revision laparoscopic fundoplication. BACKGROUND: Laparoscopic Nissen fundoplication has become the most commonly performed antireflux procedure for the treatment of gastroesophageal reflux disease, with success rates from 90 to 95%. Persistent or new symptoms often warrant endoscopic and radiographic studies to find the cause of surgical failure. In experienced hands, reoperative antireflux surgery can be done laparoscopically. We performed a retrospective analysis of all laparoscopic revision of failed fundoplications done by the principle author and the respective fellow within the laparoscopic fellowship from 1992 to 2006. METHODS: A review was performed on patients who underwent laparoscopic revision of a failed primary laparoscopic fundoplication. RESULTS: Laparoscopic revision of failed fundoplication was performed on 68 patients between 1992 and 2006. The success rate of the laparoscopic redo Nissen fundoplication was 86%. Symptoms prior to the revision procedure included heartburn (69%), dysphagia (8.8%), or both (11.7%). Preoperative evaluation revealed esophagitis in 41%, hiatal hernia with esophagitis in 36%, hiatal hernia without esophagitis in 7.3%, stenosis in 11.74%, and dysmotility in 2.4%. The main laparoscopic revisions included fundoplication alone (41%) or fundoplication with hiatal hernia repair (50%). Four gastric perforations occurred; these were repaired primarily without further incident. An open conversion was performed in 1 patient. Length of stay was 2.5 +/- 1.0 days. Mean follow-up was 22 months (range, 6-42), during which failure of the redo procedure was noted in 9 patients (13.23%). CONCLUSION: Laparoscopic redo antireflux surgery, performed in a laparoscopic fellowship program, produces excellent results that approach the success rates of primary operations. PMID- 19216694 TI - Robotic-assisted laparoscopic pyeloplasty and nephropexy for ureteropelvic junction obstruction and nephroptosis. AB - A 22-year-old female was referred with right flank pain and recurrent urinary infections. Flank pain was persistent while standing and relieved upon supine position. Intravenous urography demonstrated change of position with descent approximately 6-7 cm as the patient moved from the supine to the erect position. Diuretic renography corroborated the finding of right ureteropelvic junction obstruction (UPJO). A robotic-assisted laparoscopic dismembered pyeloplasty with simultaneous nephropexy was performed. The proximal ureter appeared to course posterior to the renal vein and then anterior to the lower pole renal artery. There was a significant "nutcracker effect" to the proximal ureter, which was causing the patient's UPJO, and the concomitant nephroptosis contributed to increase the degree of obstruction. The robotic-assisted laparoscopic pyeloplasty and nephropexy offer advantages for patients and surgeons and can be used in challenging cases with an efficacy similar to that of open repair. The robotic assisted laparoscopic pyeloplasty is the evolving standard for UPJO, especially in the presence of crossing vessels. PMID- 19216695 TI - Laparoscopic repair of perforated peptic ulcer with delayed presentation. AB - BACKGROUND: Laparoscopic repair has been used to treat perforated peptic ulcers since 1990 and is gaining acceptance. The main concerns are postoperative intra abdominal collections and performing laparoscopy in the presence of peritonitis. METHODS: From June 2006 to June 2008, 31 patients presenting with a clinical diagnosis of perforated peptic ulcer with peritonitis who underwent laparoscopic omental patch repair were selected. Patients who presented more than 24 hours after the onset of pain were only included for this analysis. Conversion criteria were malignant ulcers, perforations larger than 10 mm, or for technical difficulties. The degree of peritonitis was noted and a thorough wash given. The perforation was repaired by the standard omental patch technique, and the number of sutures were decided according to the size of the perforation. Endpoints were operative time, postoperative pain, length of postoperative hospital stay, and complications. RESULTS: There were 2 conversions due to technical difficulties. Laparoscopic repair required an average of 105 minutes to complete. The median postoperative stay was 5.5 days while mean duration of pain was 3 days. Four patients had port-site infections and 5 had intra-abdominal collections, which were managed conservatively. There were no chest infections or mortality in our series. CONCLUSIONS: Laparoscopic repair of perforated peptic ulcer is a safe, reliable procedure even in delayed presentations with peritonitis. It has an acceptable morbidity and all the advantages of the minimally invasive surgical approach. PMID- 19216696 TI - The correlation between sonographic diagnosis and laparoscopic observations on fatty liver. AB - BACKGROUND: Sonography is a comparatively cheap, convenient, and noninvasive testing on diagnosing fatty liver disease and with satisfied sensitivity and specificity. The sonographic diagnosis of fatty liver is usually made by radiologists who have few opportunities to observe the macroscopic changes on fatty liver. Having observed some obvious macroscopic changes of simple fatty liver in abdominal operations, we wanted to know how many morphological changes of the simple fatty liver there were in the end and what is the corresponding relationship between sonographic diagnosis and morphologic changes of the fatty liver. So, we designed this study for these destinations by using the intuitive and minimally invasive advantages of laparoscopy. PATIENTS AND METHODS: For this study, we chose patients with benign cholecystic disease and simple fatty liver diagnosed by sonography. In the laparoscopic operations, the surface and periphery of the liver were observed and videotaped simultaneously. Liver biopsies were performed during the operations, and the specimen was analyzed by experienced pathologists postoperatively. The videos, histologic results, and the sonographic examinations on fatty liver were compared and tried to find some potential correlation among them. RESULTS: In this study, we found some common macroscopic changes of the fatty liver under laparoscopy, such as hepatomegaly, the dullness of the liver edge, the fatty infiltration of the liver surface, the texture of the liver changing from softness to tenacity, and the blood vessels of the liver surface becoming more fuzzy. According to the histologic results, the characteristics of different gradings of fatty liver under laparoscopy could be confimed. The statistical analysis revealed a higher consistency between sonographic diagnosis and laparoscopic macroscopic changes of the fatty liver. CONCLUSION: There are some typical features of the simple fatty liver under laparoscopy. These characteristics correspond to the sonographic diagnosis on the fatty liver. There is a high consistency between sonographic diagnosis and laparoscopic findings on the fatty liver. According to the macroscopic changes of the liver under laparoscopy, the extent of simple fatty liver could be evaluated under laparoscopy. PMID- 19216697 TI - Utility of ERCP in neonatal and infant cholestasis. AB - BACKGROUND AND STUDY AIMS: In neonates and infants with biliary obstruction, the diagnosis is important to establish early. Despite the use of several noninvasive imaging modalities, a definite diagnosis of biliary atresia (BA) or other pancreaticobiliary disorders may be difficult to ascertain. In this paper, we report our experience with endoscopic retrograde cholangiopancreaticography (ERCP) in the clinical situation of neonatal and infant jaundice. PATIENTS AND METHODS: Between 1999 and 2006, we performed 23 ERCP procedures in 22 neonates and infants with the provisional diagnosis of biliary obstruction where other imaging was inconclusive. The mean age was 2.4 months and the mean weight was 4.8 kg. A pediatric videoendoscope with an outer diameter of 7.5 mm was used in all patients. RESULTS: ERCP was successful in 20 of 23 patients. BA was suggested in 6 patients, all subsequently verified by surgery. Detailed visualization of biliary structures important for planning of treatment was performed in children with choledochal cysts and spontaneous biliary perforation. Cannulation of the ampulla failed in 3 patients, of whom 1 had BA, 1 a choledochal stone, and 1 bile plug syndrome. Only 1 therapeutic ERCP was performed in a child with a stone completely obstructing the ampulla. The only complications were 1 asymptomatic case of hyperamylasemia and 1 of mild cholangitis successfully treated with antibiotics. CONCLUSIONS: ERCP is feasible and safe in the workup of neonatal cholestasis where other imaging modalities are inconclusive. Despite the expanding role of magnetic resonance cholangiography, ERCP may still have a role in the multidisciplinary workup of these patients. PMID- 19216698 TI - Laparoscopic adrenalectomy: Norwegian single-center experience of 242 procedures. AB - BACKGROUND: The last 15 years have been characterized by a rapid expansion of minimally invasive surgery as treatment for adrenal diseases. During these years, both indications and surgical techniques have shown improvements. This study analyzed an 11-year single-center experience with laparoscopic adrenalectomy. MATERIALS AND METHODS: Between January 1997 and April 2008, 242 laparoscopic adrenalectomies were performed in 220 patients at Rikshospitalet University Hospital. Of these, 192 patients were operated on for benign lesions, 23 for malignant lesions, and in 5 cases "en bloc" adrenalectomies were performed. Benign lesions included 136 hormonally active lesions (41 pheochromocytomas, 48 Conn adenomas, 25 Cushing adenomas, and 18 patients with Cushing's disease) and 56 with hormonally inactive lesions (among them, 47 nonfunctional adenomas). Malignant lesions included 16 adrenal metastases and 7 adrenocortical carcinomas. RESULTS: All adrenalectomies were completed laparoscopically. The median time of unilatateral adrenalectomy was 85 (range, 35-325) minutes. The median blood loss was 0 (range, 0-1100) mL. There were 6 intraoperative and 7 postoperative minor complications. The number of complications did not differ between the types of adrenal pathology. Only 19% of the patients required opioids postoperatively. Per and postoperative parameters were homogeneous among patients with different adrenal lesions. The patients with adrenocortical carcinoma had a distinctive intraoperative course with an evidently longer operative time and higher blood loss. The median postoperative hospital stay was 2 (range, 1-15) days. Hospital stay was the only postoperative parameter where a difference was found between patients with different adrenal lesions. The patients with carcinoma, pheochromocytoma, and Cushing's disease had the longest median postoperative stay, respectively, 5 (range, 2-6), 3 (range, 1-15), and 3 (range, 2-6) days. CONCLUSIONS: Laparoscopic adrenalectomy is a safe, effective procedure providing improved fast and uncomplicated patient recovery independent of the type of adrenal lesion. Laparoscopic adrenalectomy can be easily introduced and may soon replace traditional open surgery in specialized centers. PMID- 19216699 TI - Laparoscopic incisional and ventral hernia repair without sutures: a single center experience with 200 cases. AB - BACKGROUND: Laparoscopic incisional and ventral herniorraphy (LIVH), using a mesh, has gained recognition as an effective method and is associated with lower complication and recurrence. Controversies in the operative technique still exist about biomaterial, method of fixation, and overlap of the mesh over the defect. The aim of this study was to evaluate the outcomes achieved with LIVH in 200 consecutive patients treated in a single hospital, using fixation of the mesh with only tacks. Results of the first 100 (group A) and the last 100 (group B) operations were also compared. METHODS: From 2003 through 2007, 200 patients underwent LIVH. Overlap of the mesh was 3-5 cm. The mesh was secured with tacks alone, with the "double crown" technique. In group B, adhesiolysis was performed, avoiding high energies. RESULTS: Mean ventral defect was 107.5 (+/- 95.4) cm2. The recurrent ventral hernia rate was 20%, and the conversion rate was 2.5%. Mean operative time was 77.5 (+/- 33.9) minutes. Mean mesh dimension was 326.4 (+/- 166.8) cm2. The overall morbidity rate was 10.5%. Bowel injuries were 5 (2.5 %). Minor complications were 8.0%. Median postoperative hospital stay was 3 days. Recurrence rate was 3.5%, with a mean follow-up of 22.5 months. Chronic pain was 1%. No difference was seen between groups A and B regarding minor complications, whereas a significant difference was found regarding enterotomies (5 vs. 0; P = 0.024) and recurrences (6 vs. 1; P = 0.056). CONCLUSIONS: Fixation of the mesh with the sole use of tacks was demonstrated to be safe and effective. Avoiding high energies, no case of enterotomy occurred. PMID- 19216703 TI - Rural Palliative Care: A Comprehensive Review. AB - Abstract Background: Access to integrated, palliative care regardless of location of residence is a palliative care standard yet we know such access is limited for those living in rural and remote settings. As a beginning step in the development of research aimed at informing policy and program development, a comprehensive review of the state of knowledge regarding palliative care in rural contexts is required. Purpose: To identify, evaluate and synthesize the published literature on rural palliative care. Design: Comprehensive review. Methods: Computer searches were conducted on PubBMed, ISI Web of Science, PsycInfo, CINAHL, and Ageline using the search terms palliative care, hospice, terminal care, end-of life care, end-of-life, and rural or remote. Results: One hundred fifty-eight studies were retrieved. After screening using relevance and quality criteria, 79 studies were included in the review. Studies were grouped by subject matter into one of three categories: patient and caregiver perspectives; professional attitudes, knowledge and practice issues; and health care services. Conclusion: The body of research literature is small and eclectic, which means there is little strong evidence to inform palliative policy and service development in rural settings. Coordinated programs of research are clearly required to develop a body of knowledge that is adequate to support effective service and policy development. PMID- 19216704 TI - Emerging drugs for atopic dermatitis. AB - Atopic dermatitis (AD) is the most common chronic inflammatory skin disease, affecting 10-20% of children and 2% of adults worldwide. Preventive treatment of AD consists of daily skin hydration and emollient therapy; but the majority of patients still require symptomatic treatment with topical corticosteroids and/or topical calcineurin inhibitors, both of which may be associated with potential long-term side effects. With increasing evidence supporting the role of skin barrier defects in the pathogenesis of AD, there is also a parallel increase in medications that claim to assist barrier repair. The current review discusses some exciting results with these medications, as well as the challenges that lie ahead of them. While barrier repair treatments offer some promise, there continues to be a need for safer anti-inflammatory medications. Some of these medications under investigation are phosphodiesterase-4 inhibitors, urocanic acid oxidation products and IL-4/IL-13 receptor blockers. The review also discusses anti-staphylococcal treatments including nanocrystalline silver cream, silver and antimicrobial-coated fabrics, and anti-itch treatments including mu-opiod receptor antagonists, chymase inhibitors and cannabinoid receptor agonists. These medications may become an integral part of AD therapy. PMID- 19216705 TI - Bepotastine besilate OD tablets suppress nasal symptoms caused by Japanese cedar pollen exposure in an artificial exposure chamber (OHIO Chamber). AB - OBJECTIVE: To investigate the efficacy and safety of bepotastine, we conducted a randomized, investigator-blind, placebo-controlled and parallel study to evaluate the allergic symptoms and the cognitive function of the subjects with Japanese cedar (JC) pollinosis in response to JC pollen exposure in an artificial exposure chamber. METHODS: Twenty-four volunteers with JC pollinosis were enrolled in this study. The subjects were randomly divided into two groups: a bepotastine group and a placebo group (12 subjects each). Subjects received either a 10-mg bepotastine OD (orally disintegrating) tablet or a placebo 10 min before entering the chamber for a 3-h exposure. The nasal and ocular symptoms were evaluated by each patient at regular intervals. The amount of nasal discharge was measured and the number of sneezes recorded. To evaluate the cognitive function, a digit cancellation test (D-CAT), in which the subjects have to eliminate certain numbers within a fixed time, was conducted. RESULTS: Twelve subjects in the placebo group developed nasal and/or ocular symptoms, whereas six subjects in the bepotastine group showed none of the symptoms during exposure. Mean secretion weights and number of sneezes were significantly lower in the bepotastine group than in the placebo group. In D-CAT, no deterioration of work performance was observed in the bepotastine group. No adverse effects were observed in either group. CONCLUSION: Bepotastine treatment is effective and well tolerated in patients with allergen-induced symptoms exposed to JC pollen in an exposure chamber. PMID- 19216706 TI - Nanocolonies: detection, cloning, and analysis of individual molecules. AB - Nanocolonies (other names molecular colonies or polonies) are formed upon template nanomolecule (DNA or RNA) amplification in immobilized medium with efficient pore size in the nanometer range. This work deals with the principle, invention, development, and diverse nanocolony applications based on their unique abilities to compartmentalize amplification and expression of individual DNA and RNA molecules, including studying reactions between single molecules, digital molecular diagnostics, in vitro gene cloning and expression, as well as identification of the molecular cis-elements including DNA sequencing, analysis of single-nucleotide polymorphism, and alternative splicing investigation. PMID- 19216707 TI - Mechanisms of single-stranded DNA-binding protein functioning in cellular DNA metabolism. AB - This review deals with analysis of mechanisms involved in coordination of DNA replication and repair by SSB proteins; characteristics of eukaryotic, prokaryotic, and archaeal SSB proteins are considered, which made it possible to distinguish general mechanisms specific for functioning of proteins from organisms of different life domains. Mechanisms of SSB protein interactions with DNA during metabolism of the latter are studied; structural organization of the SSB protein complexes with DNA, as well as structural and functional peculiarities of different SSB proteins are analyzed. PMID- 19216708 TI - Bacterial 5S rRNA-binding proteins of the CTC family. AB - The presence of CTC family proteins is a unique feature of bacterial cells. In the CTC family, there are true ribosomal proteins (found in ribosomes of exponentially growing cells), and at the same time there are also proteins temporarily associated with the ribosome (they are produced by the cells under stress only and incorporate into the ribosome). One feature is common for these proteins - they specifically bind to 5S rRNA. In this review, the history of investigations of the best known representatives of this family is described briefly. Structural organization of the CTC family proteins and their occurrence among known taxonomic bacterial groups are discussed. Structural features of 5S rRNA and CTC protein are described that predetermine their specific interaction. Taking into account the position of a CTC protein and its intermolecular contacts in the ribosome, a possible role of its complex with 5S rRNA in ribosome functioning is discussed. PMID- 19216709 TI - Specific features of 5S rRNA structure - its interactions with macromolecules and possible functions. AB - Small non-coding RNAs are today a topic of great interest for molecular biologists because they can be regarded as relicts of a hypothetical "RNA world" which, apparently, preceded the modern stage of organic evolution on Earth. The small molecule of 5S rRNA (approximately 120 nucleotides) is a component of large ribosomal subunits of all living beings (5S rRNAs are not found only in mitoribosomes of fungi and metazoans). This molecule interacts with various protein factors and 23S (28S) rRNA. This review contains the accumulated data to date concerning 5S rRNA structure, interactions with other biological macromolecules, intracellular traffic, and functions in the cell. PMID- 19216710 TI - Embryonic stem cells and the problem of directed differentiation. AB - During the last two decades molecular genetic and cell mechanisms of proliferation and differentiation of mammalian stem cells have been intensively studied in leading laboratories all over the world. Studies in this field are very important both for basic science and for the development of promising cell therapy technologies. Embryonic stem cells represent a unique experimental model for the investigation of basic principles of mammalian cell differentiation and development. Using this model, important data on similarity in genetic programs during embryonic development and embryonic stem cells differentiation have been obtained. These include basically similar consequent expression of transcription factors, cell receptors, tissue specific proteins, and ion channels. Lines of embryonic stem cells are widely used for the investigation of gene functions in ontogenesis as well as in adult organisms (using gene-knockout strategy). This review deals with different pathways of mammalian (including human) embryonic stem cells differentiation. It considers the main approaches to directed differentiation of these cells in vitro: use of feeder cells, growth factors, and other chemical compounds and also genetic modification. Some examples of application of embryonic stem cells derivatives for cell therapy of some pathological conditions are discussed. PMID- 19216711 TI - Intermediate vimentin filaments and their role in intracellular organelle distribution. AB - Intermediate filaments (IF) represent one of three main cytoskeletal structures in most animal cells. The human IF protein family includes about 70 members divided into five main groups. The characteristic feature of IF is that in various cells and tissues they are formed by proteins of different groups. Structures of all IF proteins follow a unique scheme: a central alpha-helical part is flanked at the N and C ends by positively charged polypeptide chains devoid of a clear secondary structure. The central part is highly conserved for all proteins in all animals, whereas the N and C termini strongly differ both in size and amino acid composition. This review covers the broad spectrum of recent investigations of IF structure and diverse functions. Special attention is paid to the regulatory mechanisms of IF functions, mainly to phosphorylation by different protein kinases whose role is well studied. The review gives examples of hereditary diseases associated with mutations of some IF proteins, which point to an important physiological role of these cytoskeletal structures. PMID- 19216712 TI - Capping complex formation at the slow-growing end of the actin filament. AB - Actin filaments are polar; their barbed (fast-growing) and pointed (slow-growing) ends differ in structure and dynamic properties. The slow-growing end is regulated by tropomodulins, a family of capping proteins that require tropomyosins for optimal function. There are four tropomodulin isoforms; their distributions vary depending on tissue type and change during development. The C terminal half of tropomodulin contains one compact domain represented by alternating alpha-helices and beta-structures. The tropomyosin-independent actin capping site is located at the C-terminus. The N-terminal half has no regular structure; however, it contains a tropomyosin-dependent actin-capping site and two tropomyosin-binding sites. One tropomodulin molecule can bind two tropomyosin molecules. Effectiveness of tropomodulin binding to tropomyosin depends on the tropomyosin isoform. Regulation of tropomodulin binding at the pointed end as well as capping effectiveness in the presence of specific tropomyosins may affect formation of local cytoskeleton and dynamics of actin filaments in cells. PMID- 19216713 TI - Pore-forming proteins and adaptation of living organisms to environmental conditions. AB - Pore-forming proteins are powerful "tools" for adaptation of living organisms to environmental conditions. A wide range of these proteins isolated from various sources, from viruses to mammals, has been used for the analysis of their role in the processes of intra- and inter-species competition, defense, attack, and signaling. Here we review a large number of pore-forming proteins from the perspective of their functions, structures, and mechanisms of membrane penetration. Various mechanisms of cell damage, executed by these proteins in the course of formation of a pore and after its passing to conducting state, have been considered: endo- and exocytosis, lysis, necrosis, apoptosis, etc. The role of pore-forming proteins in evolution is discussed. The relevance of practical application of pore formers has been shown, including application in nanotechnological constructions. PMID- 19216714 TI - Involvement of thio-, peroxi-, and glutaredoxins in cellular redox-dependent processes. AB - Among the key antioxidant enzymes, thioredoxin and glutaredoxin systems play an important role in cell defense against oxidative stress and maintenance of redox homeostasis owing to the regulation of thiol-disulfide exchange. The thioredoxin isoforms Trx1 (cytoplasmic form) and Trx2 (mitochondrial form) can reduce inter- and intramolecular disulfide bonds in proteins, in particular, in oxidized peroxiredoxins, which disrupt organic hydroperoxides, H2O2, and peroxynitrite. NADPH-dependent thioredoxin reductase, which reduces a broad range of substrates including oxidized form of thioredoxin, can also directly reduce lipid hydroperoxides, H2O2, and dehydroascorbic and lipoic acids. Glutaredoxin, whose major isoforms in mammals are Grx1, Grx2, and Grx5, as well as thioredoxin, catalyzes S-glutathionylation and deglutathionylation of proteins to protect SH groups from oxidation and restore functionally active thiols. However, in contrast to thioredoxin, glutaredoxin reduces GSH-mixed disulfides and catalyzes the reaction not only via a dithiol mechanism but also via monothiol reduction. In addition to the role in cellular antioxidant defense, all of the reviewed redox proteins (thioredoxin, thioredoxin reductase, peroxiredoxin, and glutaredoxin) have a number of significant functions required for cell viability: they regulate transcription factor activities, play the role of growth factors, serve as enzyme cofactors, take part in regulation of cell cycle, and are involved in antiapoptotic mechanisms. PMID- 19216715 TI - D-amino acid oxidase: physiological role and applications. AB - D-Amino acids play a key role in regulation of many processes in living cells. FAD-dependent D-amino acid oxidase (DAAO) is one of the most important enzymes responsible for maintenance proper level of D-amino acids. The most interesting and important data for regulation of the nervous system, hormone secretion, and other processes by D-amino acids as well as development of different diseases under changed DAAO activity are presented. The mechanism of regulation is complex and multi-parametric because the same enzyme simultaneously influences the level of different D-amino acids, which can result in opposing effects. Use of DAAO for diagnostic and therapeutic purposes is also considered. PMID- 19216716 TI - The problem of the eukaryotic genome size. AB - The current state of knowledge concerning the unsolved problem of the huge interspecific eukaryotic genome size variations not correlating with the species phenotypic complexity (C-value enigma also known as C-value paradox) is reviewed. Characteristic features of eukaryotic genome structure and molecular mechanisms that are the basis of genome size changes are examined in connection with the C value enigma. It is emphasized that endogenous mutagens, including reactive oxygen species, create a constant nuclear environment where any genome evolves. An original quantitative model and general conception are proposed to explain the C-value enigma. In accordance with the theory, the noncoding sequences of the eukaryotic genome provide genes with global and differential protection against chemical mutagens and (in addition to the anti-mutagenesis and DNA repair systems) form a new, third system that protects eukaryotic genetic information. The joint action of these systems controls the spontaneous mutation rate in coding sequences of the eukaryotic genome. It is hypothesized that the genome size is inversely proportional to functional efficiency of the anti-mutagenesis and/or DNA repair systems in a particular biological species. In this connection, a model of eukaryotic genome evolution is proposed. PMID- 19216717 TI - Plasma membrane microdomains from hybrid aspen cells are involved in cell wall polysaccharide biosynthesis. AB - Detergent-resistant plasma membrane microdomains [DRMs (detergent-resistant membranes)] were isolated recently from several plant species. As for animal cells, a large range of cellular functions, such as signal transduction, endocytosis and protein trafficking, have been attributed to plant lipid rafts and DRMs. The data available are essentially based on proteomics and more approaches need to be undertaken to elucidate the precise function of individual populations of DRMs in plants. We report here the first isolation of DRMs from purified plasma membranes of a tree species, the hybrid aspen Populus tremula x tremuloides, and their biochemical characterization. Plasma membranes were solubilized with Triton X-100 and the resulting DRMs were isolated by flotation in sucrose density gradients. The DRMs were enriched in sterols, sphingolipids and glycosylphosphatidylinositol-anchored proteins and thus exhibited similar properties to DRMs from other species. However, they contained key carbohydrate synthases involved in cell wall polysaccharide biosynthesis, namely callose [(1- >3)-beta-D-glucan] and cellulose synthases. The association of these enzymes with DRMs was demonstrated using specific glucan synthase assays and antibodies, as well as biochemical and chemical approaches for the characterization of the polysaccharides synthesized in vitro by the isolated DRMs. More than 70% of the total glucan synthase activities present in the original plasma membranes was associated with the DRM fraction. In addition to shedding light on the lipid environment of callose and cellulose synthases, our results demonstrate the involvement of DRMs in the biosynthesis of important cell wall polysaccharides. This novel concept suggests a function of plant membrane microdomains in cell growth and morphogenesis. PMID- 19216718 TI - Development of K562 cell clones expressing beta-globin mRNA carrying the beta039 thalassaemia mutation for the screening of correctors of stop-codon mutations. AB - Nonsense mutations, giving rise to UAA, UGA and UAG stop codons within the coding region of mRNAs, promote premature translational termination and are the leading cause of approx. 30% of inherited diseases, including cystic fibrosis, Duchenne muscular dystrophy and thalassaemia. For instance, in beta(0)39-thalassaemia the CAG (glutamine) codon is mutated to the UAG stop codon, leading to premature translation termination and to mRNA destabilization through the well-described NMD (nonsense-mediated mRNA decay). In order to develop an approach facilitating translation and, therefore, protection from NMD, aminoglycoside antibiotics have been tested on mRNAs carrying premature stop codons. These drugs decrease the accuracy in the codon-anticodon base-pairing, inducing a ribosomal read-through of the premature termination codons. Interestingly, recent papers have described drugs designed and produced for suppressing premature translational termination, inducing a ribosomal read-through of premature but not normal termination codons. These findings have introduced new hopes for the development of a pharmacological approach to the therapy of beta(0)39-thalassaemia. In this context, we started the development of a cellular model of the beta(0)39-thalassaemia mutation that could be used for the screening of a high number of aminoglycosides and analogous molecules. To this aim, we produced a lentiviral construct containing the beta(0)39-thalassaemia globin gene under a minimal LCR (locus control region) control and used this construct for the transduction of K562 cells, subsequently subcloned, with the purpose to obtain several K562 clones with different integration copies of the construct. These clones were then treated with Geneticin (also known as G418) and other aminoglycosides and the production of beta-globin was analysed by FACS analysis. The results obtained suggest that this experimental system is suitable for the characterization of correction of the beta(0)39-globin mutation causing beta-thalassaemia. PMID- 19216719 TI - Low concentration thresholds of plasma membranes for rapid energy-independent translocation of a cell-penetrating peptide. AB - The exact mechanisms by which cell-penetrating peptides such as oligo-arginines and penetratin cross biological membranes has yet to be elucidated, but this is required if they are to reach their full potential as cellular delivery vectors. In the present study, qualitative and quantitative analysis of the influence of temperature, peptide concentration and plasma membrane cholesterol on the uptake and subcellular distribution of the model cell-penetrating peptide octa-arginine was performed in a number of suspension and adherent cell lines. When experiments were performed on ice, the peptide at 2 microM extracellular concentration efficiently entered and uniformly labelled the cytoplasm of all the suspension cells studied, but a 10-fold higher concentration was required to observe similar results in adherent cells. At 37 degrees C and at higher peptide concentrations, time-lapse microscopy experiments showed that the peptide rapidly penetrated the entire plasma membrane of suspension cells, with no evidence of a requirement for nucleation zones to promote this effect. Cholesterol depletion with methyl-beta cyclodextrin enhanced translocation of octa-arginine across the plasma membrane of suspension cells at 37 degrees C, but decreased overall peptide accumulation. Under the same conditions in adherent cells this agent had no effect on peptide uptake or distribution. Cholesterol depletion increased the overall accumulation of the peptide at 4 degrees C in KG1a cells, but this effect could be reversed by re-addition of cholesterol as methyl-beta-cyclodextrin-cholesterol complexes. The results highlight the relatively high porosity of the plasma membrane of suspension cells to this peptide, especially at low temperatures, suggesting that this feature could be exploited for delivering bioactive entities. PMID- 19216720 TI - Spatiotemporal activation of caspase-dependent and -independent pathways in staurosporine-induced apoptosis of p53wt and p53mt human cervical carcinoma cells. AB - BACKGROUND INFORMATION: Caspase-dependent and -independent death mechanisms are involved in apoptosis in a variety of human carcinoma cells treated with antineoplastic compounds. Our laboratory has reported that p53 is a key contributor of mitochondrial apoptosis in cervical carcinoma cells after staurosporine exposure. However, higher mitochondrial membrane potential dissipation and greater DNA fragmentation were observed in p53wt (wild-type p53) HeLa cells compared with p53mt (mutated p53) C-33A cells. Here, we have studied events linked to the mitochondrial apoptotic pathway. RESULTS: Staurosporine can induce death of HeLa cells via a cytochrome c/caspase-9/caspase-3 mitochondrial dependent apoptotic pathway and via a delayed caspase-independent pathway. In contrast with p53wt cells, p53mt C-33A cells exhibit firstly caspase-8 activation leading to caspase-3 activation and Bid cleavage followed by cytochrome c release. Attenuation of PARP-1 [poly(ADP-ribose) polymerase-1] cleavage as well as oligonucleosomal DNA fragmentation in the presence of z-VAD-fmk points toward a major involvement of a caspase-dependent pathway in staurosporine-induced apoptosis in p53wt HeLa cells, which is not the case in p53mt C-33A cells. Meanwhile, the use of 3-aminobenzamide, a PARP-1 inhibitor known to prevent AIF (apoptosis-inducing factor) release, significantly decreases staurosporine induced death in these p53mt carcinoma cells, suggesting a preferential implication of caspase-independent apoptosis. On the other hand, we show that p53, whose activity is modulated by pifithrin-alpha, isolated as a suppressor of p53-mediated transactivation, or by PRIMA-1 (p53 reactivation and induction of massive apoptosis), that reactivates mutant p53, causes cytochrome c release as well as mitochondrio-nuclear AIF translocation in staurosporine-induced apoptosis of cervical carcinoma cells. CONCLUSIONS: The present paper highlights that staurosporine engages the intrinsic mitochondrial apoptotic pathway via caspase-8 or caspase-9 signalling cascades and via caspase-independent cell death, as well as through p53 activity. PMID- 19216721 TI - The effect of antimicrobial resistance on patient outcomes: importance of proper evaluation of appropriate therapy. AB - The impact of antimicrobial resistance on patient outcomes can be effectively measured only if the appropriateness of the antimicrobial therapy received is properly measured. Definition of appropriate therapy should include not only in vitro susceptibility but also the clinical adequacy of the antibiotic used, taking into account the pathogen isolated, the site of infection, known pharmacokinetic and pharmacodynamic properties of the drug, and dosing. In the absence of these data, the effect of delay or absence of appropriate therapy in patients infected with resistant bacterial pathogens is subject to confounding, and the true effect of resistance on outcomes may be obscured. PMID- 19216722 TI - Lymphocytes, apoptosis and sepsis: making the jump from mice to humans. AB - Sepsis is an important clinical problem with a mortality rate of 20% to 30%. Lymphocyte apoptosis has been recognized as an important step in the pathogenesis of experimental sepsis, by inducing a state of 'immune paralysis' that renders the host vulnerable to invading pathogens. The importance of lymphocyte apoptosis in human disease is now confirmed by Weber and colleagues, who demonstrate extensive apoptosis in circulating lymphocytes from patients with severe sepsis. Weber and colleagues' data set the basis for further studies aimed at modulating lymphocyte apoptosis in sepsis. PMID- 19216723 TI - Should dialysis be offered in all cases of metformin-associated lactic acidosis? AB - Metformin is commonly used in diabetes mellitus type 2, with lactic acidosis being a rare but potentially fatal complication of this therapy. The management of metformin-associated lactic acidosis (MALA) is controversial. Treatment may include supportive care, activated charcoal, bicarbonate infusion, hemodialysis, or continuous venovenous hemofiltration. In the previous issue of Critical Care, Peters and colleagues systematically evaluated outcomes in MALA patients admitted to their intensive care unit. The mortality rate of patients who received dialysis was similar to that of patients who were not dialyzed. However, it was the more acutely and chronically ill patients who actually received dialysis. This suggests that hemodialysis was beneficial in preventing a higher mortality rate in those who required renal replacement therapy. PMID- 19216726 TI - Sperm dumping as a defense against meiotic drive. AB - Sperm from Drosophila simulans that carry a sex-ratio distorter is preferentially lost from females' sperm-storage organs. This suggests that sperm dumping is a major factor affecting sperm competition in this species, and may have evolved in response to sex-ratio distorters. PMID- 19216724 TI - Bench-to-bedside review: ventilatory abnormalities in sepsis. AB - In septic patients increased central drive and increased metabolic demands combine to increase energy demands on the ventilatory muscles. This occurs at a time when energy supplies are limited and energy production hindered, and it leads to an energy supply-demand imbalance and often ventilatory failure. Problems related to contractile function of the ventilatory muscles also contribute, especially when the clinical course is prolonged. The increased ventilatory activity increases interactions between the ventilatory and cardiovascular systems, and when ventilatory muscles fail and mechanical ventilatory support is required a new set of problems emerges. In this review I discuss factors related to ventilatory muscle failure, giving emphasis to mechanical and supply demand aspects. I also review the implications of changes in ventilatory patterns for heart-lung interactions. PMID- 19216727 TI - Are we HER-ting for innovation in neoadjuvant breast cancer trial design? AB - Through the use of surrogate markers of efficacy, neoadjuvant studies may facilitate the implementation of new treatments into clinical practice. However, disease-free survival is the current standard outcome endpoint for registration of a novel treatment. The coupling of smaller neoadjuvant 'proof of principle' studies with larger adjuvant registration trials offers the promise of speeding up the time to market of new therapies. Clever new designs, such as the 'biological window' and 'learn on the way', can provide valuable insight regarding mechanisms of action and resistance of these novel drugs by identifying patients who are most likely to respond to a novel therapy early in the drug development process. Using the ongoing neoadjuvant trials with HER2 (human epidermal growth factor receptor 2)-directed therapy as a paradigm, this article discusses recent innovations in study design and the challenges of conducting translational research in the neoadjuvant setting. PMID- 19216728 TI - Happy together: genomic insights into the unique Nanoarchaeum/Ignicoccus association. AB - The complete genome sequence of the crenarchaeon Ignicoccus hospitalis published recently in Genome Biology provides a great leap forward in the dissection of its unique association with another hyperthermophilic archaeon, Nanoarchaeum equitans. PMID- 19216729 TI - Cross-talk in the gut. AB - Modulation of host signaling by the products of microbial activity in the gut may affect weight gain and fat formation. PMID- 19216730 TI - Copy number variation goes clinical. AB - A report of the First Golden Helix Symposium 'Copy Number Variation (CNV) and Genomic Alterations in Health and Disease', Athens, Greece, 28-29 November 2008. PMID- 19216731 TI - Genetic control of disease in an experimental model for Sjogren's syndrome. AB - Sjogren's syndrome is an autoimmune disease with a complex etiology depending on hereditary and environmental factors. The disease is characterized by lymphocytic infiltration and inflammation in the salivary and lacrimal glands, leading to oral and ocular dryness. To understand the genetic susceptibility in Sjogren's syndrome, studies of disease phenotypes have been performed in the non-obese diabetic (NOD) mouse. By the identification of genetic regions controlling development of autoimmune exocrinopathy in the NOD mouse and by reducing one of these regions considerably, Nguyen et al. in a recent issue of Arthritis Research and Therapy propose candidate genes for development of Sjogren's syndrome. PMID- 19216732 TI - Lateral inhibition and concentration-invariant odor perception. AB - Sensory identity usually remains constant across a large intensity range. Vertebrates use lateral inhibition to match the sensitivity of retinal ganglion cells to the intensity of light. A new study published in Journal of Biology suggests that lateral inhibition in the Drosophila antennal lobe is similarly required for concentration-invariant perception of odors. PMID- 19216733 TI - Diagnosis and treatment delay among pulmonary tuberculosis patients identified using the Taiwan reporting enquiry system, 2002-2006. AB - BACKGROUND: The tuberculosis reporting enquiry system was launched in Taiwan in 2001. Tuberculosis has been categorized as the third most important notifiable disease in Taiwan and the time required for reporting has been shortened to 7 days. METHODS: A total of 114,827 cases were reported using the Taiwan enquiry system between 2002 and 2006; of these, 26,027 (22.7%) were finally diagnosed as not being tuberculosis, 7,005 (8.2%) were diagnosed as extra-pulmonary tuberculosis and 3,677 (3.2%) were not a first-time diagnosis of tuberculosis, and these cases were hence excluded. Diagnosis time was defined as the length of time between the first medical examination (including chest radiography, sputum smear or sputum culture) to the diagnosis of PTB; treatment time was defined as the period from the diagnosis of PTB to the initiation of treatment. Using the cut-off at the 75th percentile, a period of longer than 9 days was defined as a diagnosis delay and a period of longer than 2 days as a treatment delay. Multiple logistic regression analysis was applied to analyze the risk factors associated with these delays. RESULTS: During the five-year study period, among the 78,118 new PTB patients reported in Taiwan, the mean diagnosis and treatment times were 12 and 5 days and the median times 1 day and 0 days, respectively. In total, 24.9% of the new PTB patients' diagnosis time delays were longer than 9 days and 20.3% of the patients' treatment time delays were longer than 2 days. The main factors associated with diagnosis delay included age, reporting year, living with family and a positive sputum culture (p < 0.0001); the risk factors significantly associated with treatment delay were increased age, an aboriginal ethnic background, a positive sputum culture and diagnosis at a non-medical center (p < 0.0001). CONCLUSION: The Taiwan TB reporting enquiry system has successfully increased the confirmed PTB reporting rate from 64.4% to 71.5%. Greater age and a positive sputum culture were both found to significantly increase both diagnosis and treatment delays; treatment delay is also significantly affected by the patient having an aboriginal ethnic background and being diagnosed at a non medical center. PMID- 19216734 TI - A multi-center, randomized, clinical study to compare the effect and safety of autologous cultured osteoblast(Ossron) injection to treat fractures. AB - BACKGROUND: We performed a multicenter, open, randomized, clinical study of autologous cultured osteoblast injection for long-bone fracture, to evaluate the fracture healing acceleration effect and the safety of autologous cultured osteoblasts. METHODS: Sixty-four patients with long-bone fractures were randomly divided into two groups, i.e. those who received autologous cultured osteoblast injection and those who received no treatment. The sum of the difference in the callus formation scores after four and eight weeks, was used as the first efficacy variable. RESULTS: The autologous cultured osteoblast injection group showed fracture healing acceleration of statistical significance, and there were no specific patient complications when using this treatment. CONCLUSION: Autologous cultured osteoblast injection should therefore be considered as a successful treatment option for treating long-bone fracture. PMID- 19216735 TI - Loss of full length CtBP1 expression enhances the invasive potential of human melanoma. AB - BACKGROUND: The C-terminal binding protein 1 (CtBP1) is a known co-repressor of gene transcription. We recently revealed that CtBP1 expression is lost in melanoma cells and melanoma inhibitory activity (MIA) expression is subsequently increased. The present study was performed to evaluate a more general role of CtBP1 in human melanoma and identify further CtBP1-regulated target genes. METHODS: Sequence analysis and expression profile of CtBP1 in melanoma cell lines were done by PCR. Boyden Chamber assays and co-immunoprecipitation were performed to investigate the functional role of CtBP1. Gene expression analysis and micro array data were used to define target genes. RESULTS: Interestingly, we detected an alternative splice product of CtBP1 with unknown function whose expression is induced at reduction of full length CtBP1. Overexpression of full length CtBP1 in melanoma cells had no effect on cell proliferation but did influence cell migration and invasiveness. To understand the effect of CtBP1 we identified putative LEF/TCF target genes found to be strongly expressed in melanoma using DNA microarray analysis. We focused on fourteen genes not previously associated with melanoma. Detailed analysis revealed that most of these were known to be involved in tumor metastasis. Eleven genes had expression profiles associated with melanoma cell invasiveness. CONCLUSION: In summary, this study revealed that reduction of CtBP1 expression is correlated with migratory, invasive potential of melanoma cells. PMID- 19216736 TI - Comprehensive characterization of neuroblastoma cell line subtypes reveals bilineage potential similar to neural crest stem cells. AB - BACKGROUND: Neuroblastic tumors (NBT) derive from neural crest stem cells (NCSC). Histologically, NBT are composed by neuroblasts and Schwannian cells. In culture, neuroblastic (N-), substrate-adherent (S-) and intermediate phenotype (I-) cell subtypes arise spontaneously. METHODS: Here, neuroblastoma (NB) cell line subtypes were characterized according to embryonic peripheral nervous system development markers (GAP43, Phox2b, Sox10, c-kit, GD2, NF68, vimentin, S100beta, calcyclin and ABCG2), morphological features, gene expression and differentiation potential. I-type cells were investigated as a bipotential (neuronal and glial) differentiation stage. RESULTS: Positive immunostaining of NCSC (GAP43, c-kit, NF68, vimentin and Phox2b) and undifferentiated cell (ABCG2) markers was observed in all NB subtypes. N- and I-type cells displayed cytoplasmic membrane GD2 staining, while nuclear calcyclin was restricted to S-type. N- and I-type cells showed similar phenotype and immunoreactivity pattern. Differential gene expression was associated with each cell subtype. N- and I-type cells displayed similar differentiation capacity towards neuronal and glial lineage fates. S-type cells, upon induction, did not show a neuronal-like phenotype, despite gene expression changes. CONCLUSION: Results suggest that N- and I-type NB cell subtypes represent an immature bilineage stage, able to progress towards neuronal and glial fates upon induction of differentiation. S-type cells appear irreversibly committed to a glial lineage fate. PMID- 19216737 TI - Enhanced pre-synaptic glutamate release in deep-dorsal horn contributes to calcium channel alpha-2-delta-1 protein-mediated spinal sensitization and behavioral hypersensitivity. AB - Nerve injury-induced expression of the spinal calcium channel alpha-2-delta-1 subunit (Cavalpha2delta1) has been shown to mediate behavioral hypersensitivity through a yet identified mechanism. We examined if this neuroplasticity modulates behavioral hypersensitivity by regulating spinal glutamatergic neurotransmission in injury-free transgenic mice overexpressing the Cavalpha2delta1 proteins in neuronal tissues. The transgenic mice exhibited hypersensitivity to mechanical stimulation (allodynia) similar to the spinal nerve ligation injury model. Intrathecally delivered antagonists for N-methyl-D-aspartate (NMDA) and alpha amino-3-hydroxyl-5-methylisoxazole-4-propionic acid (AMPA)/kainate receptors, but not for the metabotropic glutamate receptors, caused a dose-dependent allodynia reversal in the transgenic mice without changing the behavioral sensitivity in wild-type mice. This suggests that elevated spinal Cavalpha2delta1 mediates allodynia through a pathway involving activation of selective glutamate receptors. To determine if this is mediated by enhanced spinal neuronal excitability or pre-synaptic glutamate release in deep-dorsal horn, we examined wide-dynamic-range (WDR) neuron excitability with extracellular recording and glutamate-mediated excitatory postsynaptic currents with whole-cell patch recording in deep-dorsal horn of the Cavalpha2delta1 transgenic mice. Our data indicated that overexpression of Cavalpha2delta1 in neuronal tissues led to increased frequency, but not amplitude, of miniature excitatory post synaptic currents mediated mainly by AMPA/kainate receptors at physiological membrane potentials, and also by NMDA receptors upon depolarization, without changing the excitability of WDR neurons to high intensity stimulation. Together, these findings support a mechanism of Cavalpha2delta1-mediated spinal sensitization in which elevated Cavalpha2delta1 causes increased pre-synaptic glutamate release that leads to reduced excitation thresholds of post-synaptic dorsal horn neurons to innocuous stimuli. This spinal sensitization mechanism may mediate at least partially the neuropathic pain states derived from increased pre-synaptic Cavalpha2delta1 expression. PMID- 19216738 TI - Healthcare workers' attitudes to working during pandemic influenza: a qualitative study. AB - BACKGROUND: Healthcare workers (HCWs) will play a key role in any response to pandemic influenza, and the UK healthcare system's ability to cope during an influenza pandemic will depend, to a large extent, on the number of HCWs who are able and willing to work through the crisis. UK emergency planning will be improved if planners have a better understanding of the reasons UK HCWs may have for their absenteeism, and what might motivate them to work during an influenza pandemic.This paper reports the results of a qualitative study that explored UK HCWs' views (n = 64) about working during an influenza pandemic, in order to identify factors that might influence their willingness and ability to work and to identify potential sources of any perceived duty on HCWs to work. METHODS: A qualitative study, using focus groups (n = 9) and interviews (n = 5). RESULTS: HCWs across a range of roles and grades tended to feel motivated by a sense of obligation to work through an influenza pandemic. A number of significant barriers that may prevent them from doing so were also identified. Perceived barriers to the ability to work included being ill oneself, transport difficulties, and childcare responsibilities. Perceived barriers to the willingness to work included: prioritising the wellbeing of family members; a lack of trust in, and goodwill towards, the NHS; a lack of information about the risks and what is expected of them during the crisis; fear of litigation; and the feeling that employers do not take the needs of staff seriously. Barriers to ability and barriers to willingness, however, are difficult to separate out. CONCLUSION: Although our participants tended to feel a general obligation to work during an influenza pandemic, there are barriers to working, which, if generalisable, may significantly reduce the NHS workforce during a pandemic. The barriers identified are both barriers to willingness and to ability. This suggests that pandemic planning needs to take into account the possibility that staff may be absent for reasons beyond those currently anticipated in UK planning documents. In particular, staff who are physically able to attend work may nonetheless be unwilling to do so. Although there are some barriers that cannot be mitigated by employers (such as illness, transport infrastructure etc.), there are a number of remedial steps that can be taken to lesson the impact of others (providing accommodation, building reciprocity, provision of information and guidance etc). We suggest that barriers to working lie along an ability/willingness continuum, and that absenteeism may be reduced by taking steps to prevent barriers to willingness becoming perceived barriers to ability. PMID- 19216739 TI - The role of RNA folding free energy in the evolution of the polymerase genes of the influenza A virus. AB - BACKGROUND: The influenza A virus genome is composed of eight single-stranded RNA segments of negative polarity. Although the hemagglutinin and neuraminidase genes are known to play a key role in host adaptation, the polymerase genes (which encode the polymerase segments PB2, PB1, PA) and the nucleoprotein gene are also important for the efficient propagation of the virus in the host and for its adaptation to new hosts. Current efforts to understand the host-specificity of the virus have largely focused on the amino acid differences between avian and human isolates. RESULTS: Here we show that the folding free energy of the RNA segments may play an equally important role in the evolution and host adaptation of the influenza virus. Folding free energy may affect the stability of the viral RNA and influence the rate of viral protein translation. We found that there is a clear distinction between the avian and human folding free energy distributions for the polymerase and the nucleoprotein genes, with human viruses having substantially higher folding free energy values. This difference is independent of the amino acid composition and the codon bias. Furthermore, the folding free energy values of the commonly circulating human viruses tend to shift towards higher values over the years, after they entered the human population. Finally, our results indicate that the temperature in which the cells grow affects infection efficiency. CONCLUSIONS: Our data suggest for the first time that RNA structure stability may play an important role in the emergence and host shift of influenza A virus. The fact that cell temperature affects virus propagation in mammalian cells could help identify those avian strains that pose a higher threat to humans. PMID- 19216740 TI - A module-based analytical strategy to identify novel disease-associated genes shows an inhibitory role for interleukin 7 Receptor in allergic inflammation. AB - BACKGROUND: The identification of novel genes by high-throughput studies of complex diseases is complicated by the large number of potential genes. However, since disease-associated genes tend to interact, one solution is to arrange them in modules based on co-expression data and known gene interactions. The hypothesis of this study was that such a module could be a) found and validated in allergic disease and b) used to find and validate one ore more novel disease associated genes. RESULTS: To test these hypotheses integrated analysis of a large number of gene expression microarray experiments from different forms of allergy was performed. This led to the identification of an experimentally validated reference gene that was used to construct a module of co-expressed and interacting genes. This module was validated in an independent material, by replicating the expression changes in allergen-challenged CD4+ cells. Moreover, the changes were reversed following treatment with corticosteroids. The module contained several novel disease-associated genes, of which the one with the highest number of interactions with known disease genes, IL7R, was selected for further validation. The expression levels of IL7R in allergen challenged CD4+ cells decreased following challenge but increased after treatment. This suggested an inhibitory role, which was confirmed by functional studies. CONCLUSION: We propose that a module-based analytical strategy is generally applicable to find novel genes in complex diseases. PMID- 19216741 TI - Validity of the diagnosis of a single depressive episode in a case register. AB - OBJECTIVE: To validate the ICD-10 diagnosis of a single depressive episode as used in daily clinical psychiatric practice and as recorded in the Danish Psychiatric Central Research Register. METHODS: Patients discharged with a diagnosis of a single depressive episode were consecutively sampled from the register and diagnosed according to an interview using the Schedules for Clinical Assessment in Neuropsychiatry (SCAN). RESULTS: A total of 75.4% of 399 patients with a register diagnosis of a single depressive episode also got this diagnosis according to the SCAN interview (82.8% for severe type of a single depression, 76.0% for moderate type of a single depression and 65.2% for mild type of a single depression). CONCLUSION: The ICD-10 diagnosis of a single depressive episode can be used in daily clinical practice with sufficient precision. The validity of the diagnosis is highest for severe and moderate type of depression and decreases for mild depression. PMID- 19216742 TI - Gene expression in primate liver during viral hemorrhagic fever. AB - BACKGROUND: Rhesus macaques infected with lymphocytic choriomeningitis virus (LCMV) provide a model for human Lassa fever. Disease begins with flu-like symptoms and progresses rapidly with fatal consequences. Previously, we profiled the blood transcriptome of LCMV-infected monkeys (M. Djavani et al J. Virol. 2007) showing distinct pre-viremic and viremic stages that discriminated virulent from benign infections. In the present study, changes in liver gene expression from macaques infected with virulent LCMV-WE were compared to gene expression in uninfected monkeys as well as to monkeys that were infected but not diseased. RESULTS: Based on a functional pathway analysis of differentially expressed genes, virulent LCMV-WE had a broader effect on liver cell function than did infection with non-virulent LCMV-Armstrong. During the first few days after infection, LCMV altered expression of genes associated with energy production, including fatty acid and glucose metabolism. The transcriptome profile resembled that of an organism in starvation: mRNA for acetyl-CoA carboxylase, a key enzyme of fatty acid synthesis was reduced while genes for enzymes in gluconeogenesis were up-regulated. Expression was also altered for genes associated with complement and coagulation cascades, and with signaling pathways involving STAT1 and TGF-beta. CONCLUSION: Most of the 4500 differentially expressed transcripts represented a general response to both virulent and mild infections. However, approximately 250 of these transcripts had significantly different expression in virulent infections as compared to mild infections, with approximately 30 of these being differentially regulated during the pre-viremic stage of infection. The genes that are expressed early and differently in mild and virulent disease are potential biomarkers for prognosis and triage of acute viral disease. PMID- 19216743 TI - A simple model for the early events of quorum sensing in Pseudomonas aeruginosa: modeling bacterial swarming as the movement of an "activation zone". AB - BACKGROUND: Quorum sensing (QS) is a form of gene regulation based on cell density that depends on inter-cellular communication. While there are a variety of models for bacterial colony morphology, there is little work linking QS genes to movement in an open system. RESULTS: The onset of swarming in environmental P. aeruginosa PUPa3 was described with a simplified computational model in which cells in random motion communicate via a diffusible signal (representing N-acyl homoserine lactones, AHL) as well as diffusible, secreted factors (enzymes, biosurfactans, i.e. "public goods") that regulate the intensity of movement and metabolism in a threshold-dependent manner. As a result, an "activation zone" emerges in which nutrients and other public goods are present in sufficient quantities, and swarming is the spontaneous displacement of this high cell density zone towards nutrients and/or exogenous signals. The model correctly predicts the behaviour of genomic knockout mutants in which the QS genes responsible either for the synthesis (lasI, rhlI) or the sensing (lasR, rhlR) of AHL signals were inactivated. For wild type cells the model predicts sustained colony growth that can however be collapsed by the overconsumption of nutrients. CONCLUSION: While in more complex models include self-orienting abilities that allow cells to follow concentration gradients of nutrients and chemotactic agents, in this model, displacement towards nutrients or environmental signals is an emergent property of the community that results from the action of a few, well defined QS genes and their products. Still the model qualitatively describes the salient properties of QS bacteria, i.e. the density-dependent onset of swarming as well as the response to exogenous signals or cues. PMID- 19216744 TI - Chemical tools selectively target components of the PKA system. AB - BACKGROUND: In the eukaryotic cell the cAMP-dependent protein kinase (PKA) is a key enzyme in signal transduction and represents the main target of the second messenger cAMP. Here we describe the design, synthesis and characterisation of specifically tailored cAMP analogs which can be utilised as a tool for affinity enrichment and purification as well as for proteomics based analyses of cAMP binding proteins. RESULTS: Two sets of chemical binders were developed based on the phosphorothioate derivatives of cAMP, Sp-cAMPS and Rp-cAMPS acting as cAMP agonists and -antagonists, respectively. These compounds were tested via direct surface plasmon resonance (SPR) analyses for their binding properties to PKA R subunits and holoenzyme. Furthermore, these analogs were used in an affinity purification approach to analyse their binding and elution properties for the enrichment and improvement of cAMP binding proteins exemplified by the PKA R subunits. As determined by SPR, all tested Sp-analogs provide valuable tools for affinity chromatography. However, Sp-8-AEA-cAMPS displayed (i) superior enrichment properties while maintaining low unspecific binding to other proteins in crude cell lysates, (ii) allowing mild elution conditions and (iii) providing the capability to efficiently purify all four isoforms of active PKA R-subunit in milligram quantities within 8 h. In a chemical proteomics approach both sets of binders, Rp- and Sp-cAMPS derivatives, can be employed. Whereas Sp-8-AEA-cAMPS preferentially binds free R-subunit, Rp-AHDAA-cAMPS, displaying antagonist properties, not only binds to the free PKA R-subunits but also to the intact PKA holoenzyme both from recombinant and endogenous sources. CONCLUSION: In summary, all tested cAMP analogs were useful for their respective application as an affinity reagent which can enhance purification of cAMP binding proteins. Sp-8 AEA-cAMPS was considered the most efficient analog since Sp-8-AHA-cAMPS and Sp-2 AHA-cAMPS, demonstrated incomplete elution from the matrix, as well as retaining notable amounts of bound protein contaminants. Furthermore it could be demonstrated that an affinity resin based on Rp-8-AHDAA-cAMPS provides a valuable tool for chemical proteomics approaches. PMID- 19216745 TI - Etiological explanation, treatability and preventability of childhood autism: a survey of Nigerian healthcare workers' opinion. AB - BACKGROUND: Because of their peculiar sociocultural background, healthcare workers in sub-Saharan African subcultures may have various conceptions on different aspects of autism spectrum disorders (ASD), such as etiology, treatment and issues of prognosis. These various conceptions, if different from current knowledge in literature about ASD, may negatively influence help-seeking behavior of parents of children with ASD who seek advice and information from the healthcare workers. This study assessed the opinions of healthcare workers in Nigeria on aspects of etiology, treatability and preventability of childhood autism, and relates their opinions to the sociodemographic variables. METHODS: Healthcare workers working in four tertiary healthcare facilities located in the south-east and south-south regions of Nigeria were interviewed with a sociodemographic questionnaire, personal opinion on etiology, treatability and preventability of childhood autism (POETPCA) questionnaire and knowledge about childhood autism among health workers (KCAHW) questionnaire to assess their knowledge and opinions on various aspects of childhood autism. RESULTS: A total of 134 healthcare workers participated in the study. In all, 78 (58.2%), 19 (14.2%) and 36 (26.9%) of the healthcare workers were of the opinion that the etiology of childhood autism can be explained by natural, preternatural and supernatural causes, respectively. One (0.7%) of the healthcare workers was unsure of the explanation of the etiology. Knowledge about childhood autism as measured by scores on the KCAHW questionnaire was the only factor significantly associated with the opinions of the healthcare workers on etiology of childhood autism. In all, 73 (54.5%) and 43 (32.1%), of the healthcare workers subscribed to the opinion that childhood autism is treatable and preventable respectively. Previous involvement with managing children with ASD significantly influenced the opinion of the healthcare workers in subscribing to treatability of childhood autism, while working experience of less than 6 years among the healthcare workers significantly influenced the opinion of the healthcare workers in admitting to believing in the preventability of childhood autism. CONCLUSION: In designing policies and programs to change negative opinions or beliefs of healthcare workers about childhood autism, there is a need for baseline information such as this survey. Changing the negative opinions or beliefs of the healthcare workers about childhood autism should encourage appropriate help seeking behavior among parents of children with ASD who may be seeking advice or information from the healthcare workers. This would encourage early interventions, which are essential to prognosis of childhood autism. PMID- 19216746 TI - Gain and loss of elongation factor genes in green algae. AB - BACKGROUND: Two key genes of the translational apparatus, elongation factor-1 alpha (EF-1alpha) and elongation factor-like (EFL) have an almost mutually exclusive distribution in eukaryotes. In the green plant lineage, the Chlorophyta encode EFL except Acetabularia where EF-1alpha is found, and the Streptophyta possess EF-1alpha except Mesostigma, which has EFL. These results raise questions about evolutionary patterns of gain and loss of EF-1alpha and EFL. A previous study launched the hypothesis that EF-1alpha was the primitive state and that EFL was gained once in the ancestor of the green plants, followed by differential loss of EF-1alpha or EFL in the principal clades of the Viridiplantae. In order to gain more insight in the distribution of EF-1alpha and EFL in green plants and test this hypothesis we screened the presence of the genes in a large sample of green algae and analyzed their gain-loss dynamics in a maximum likelihood framework using continuous-time Markov models. RESULTS: Within the Chlorophyta, EF-1alpha is shown to be present in three ulvophycean orders (i.e., Dasycladales, Bryopsidales, Siphonocladales) and the genus Ignatius. Models describing gene gain-loss dynamics revealed that the presence of EF-1alpha, EFL or both genes along the backbone of the green plant phylogeny is highly uncertain due to sensitivity to branch lengths and lack of prior knowledge about ancestral states or rates of gene gain and loss. Model refinements based on insights gained from the EF-1alpha phylogeny reduce uncertainty but still imply several equally likely possibilities: a primitive EF-1alpha state with multiple independent EFL gains or coexistence of both genes in the ancestor of the Viridiplantae or Chlorophyta followed by differential loss of one or the other gene in the various lineages. CONCLUSION: EF-1alpha is much more common among green algae than previously thought. The mutually exclusive distribution of EF-1alpha and EFL is confirmed in a large sample of green plants. Hypotheses about the gain-loss dynamics of elongation factor genes are hard to test analytically due to a relatively flat likelihood surface, even if prior knowledge is incorporated. Phylogenetic analysis of EFL genes indicates misinterpretations in the recent literature due to uncertainty regarding the root position. PMID- 19216747 TI - Human Papillomaviruses and genital co-infections in gynaecological outpatients. AB - BACKGROUND: High grade HPV infections and persistence are the strongest risk factors for cervical cancer. Nevertheless other genital microorganisms may be involved in the progression of HPV associated lesions. METHODS: Cervical samples were collected to search for human Papillomavirus (HPV), bacteria and yeast infections in gynaecologic outpatients. HPV typing was carried out by PCR and sequencing on cervical brush specimens. Chlamydia trachomatis was identified by strand displacement amplification (SDA) and the other microorganisms were detected by conventional methods. RESULTS: In this cross-sectional study on 857 enrolled outpatients, statistical analyses revealed a significant association of HPV with C. trachomatis and Ureaplasma urealyticum (at high density) detection, whereas no correlation was found between HPV infection and bacterial vaginosis, Streptococcus agalactiae, yeasts, Trichomonas vaginalis and U. urealyticum. Mycoplasma hominis was isolated only in a few cases both in HPV positive and negative women and no patient was infected with Neisseria gonorrhoeae. CONCLUSION: Although bacterial vaginosis was not significantly associated with HPV, it was more common among the HPV positive women. A significant association between HPV and C. trachomatis was found and interestingly also with U. urealyticum but only at a high colonization rate. These data suggest that it may be important to screen for the simultaneous presence of different microorganisms which may have synergistic pathological effects. PMID- 19216748 TI - NF-kappaB activation by Helicobacter pylori requires Akt-mediated phosphorylation of p65. AB - BACKGROUND: The inflammatory response in Helicobacter pylori-infected gastric tissue is mediated by cag pathogenicity island (PAI)-dependent activation of nuclear factor-kappaB (NF-kappaB). Phosphatidylinositol 3-kinase (PI3K)/Akt signaling is known to play a role in NF-kappaB activation, but little information is available on the relationship between H. pylori and PI3K/Akt signaling in gastric epithelial cells. We examined whether H. pylori activates Akt in gastric epithelial cells, the role of cag PAI in this process and the role of Akt in regulating H. pylori-induced NF-kappaB activation. RESULTS: Phosphorylated Akt was detected in epithelial cells of H. pylori-positive gastric tissues. Although Akt was activated in MKN45 and AGS cells by coculture with cag PAI-positive H. pylori strains, a cag PAI-negative mutant showed no activation of Akt. H. pylori also induced p65 phosphorylation. PI3K inhibitor suppressed H. pylori-induced p65 phosphorylation and NF-kappaB transactivation, as well as interleukin-8 expression. Furthermore, transfection with a dominant-negative Akt inhibited H. pylori-induced NF-kappaB transactivation. Transfection with small interference RNAs for p65 and Akt also inhibited H. pylori-induced interleukin-8 expression. CONCLUSION: The results suggest that cag PAI-positive H. pylori activates Akt in gastric epithelial cells and this may contribute to H. pylori-mediated NF-kappaB activation associated with mucosal inflammation and carcinogenesis. PMID- 19216749 TI - Prolonging life and delaying death: the role of physicians in the context of limited intensive care resources. AB - Critical care is in an emerging crisis of conflict between what individuals expect and the economic burden society and government are prepared to provide. The goal of critical care support is to prevent suffering and premature death by intensive therapy of reversible illnesses within a reasonable timeframe. Recently, it has become apparent that early support in an intensive care environment can improve patient outcomes. However, life support technology has advanced, allowing physicians to prolong life (and postpone death) in circumstances that were not possible in the recent past. This has been recognized by not only the medical community, but also by society at large. One corollary may be that expectations for recovery from critical illness have also become extremely high. In addition, greater numbers of patients are dying in intensive care units after having receiving prolonged durations of life-sustaining therapy. Herein lies the emerging crisis -- critical care therapy must be available in a timely fashion for those who require it urgently, yet its provision is largely dependent on a finite availability of both capital and human resources. Physicians are often placed in a troubling conflict of interest by pressures to use health resources prudently while also promoting the equitable and timely access to critical care therapy. In this commentary, these issues are broadly discussed from the perspective of the individual clinician as well as that of society as a whole. The intent is to generate dialogue on the dynamic between individual clinicians navigating the complexities of how and when to use critical care support in the context of end-of-life issues, the increasing demands placed on finite critical care capacity, and the reasonable expectations of society. PMID- 19216750 TI - Asymmetric relationships between proteins shape genome evolution. AB - BACKGROUND: The relationships between proteins are often asymmetric: one protein (A) depends for its function on another protein (B), but the second protein does not depend on the first. In metabolic networks there are multiple pathways that converge into one central pathway. The enzymes in the converging pathways depend on the enzymes in the central pathway, but the enzymes in the latter do not depend on any specific enzyme in the converging pathways. Asymmetric relations are analogous to the "if->then" logical relation where A implies B, but B does not imply A (A->B). RESULTS: We show that the majority of relationships between enzymes in metabolic flux models of metabolism in Escherichia coli and Saccharomyces cerevisiae are asymmetric. We show furthermore that these asymmetric relationships are reflected in the expression of the genes encoding those enzymes, the effect of gene knockouts and the evolution of genomes. From the asymmetric relative dependency, one would expect that the gene that is relatively independent (B) can occur without the other dependent gene (A), but not the reverse. Indeed, when only one gene of an A->B pair is expressed, is essential, is present in a genome after an evolutionary gain or loss, it tends to be the independent gene (B). This bias is strongest for genes encoding proteins whose asymmetric relationship is evolutionarily conserved. CONCLUSIONS: The asymmetric relations between proteins that arise from the system properties of metabolic networks affect gene expression, the relative effect of gene knockouts and genome evolution in a predictable manner. PMID- 19216751 TI - Identification and quantification of prosthetic mitral regurgitation by flow convergence method using transthoracic approach. AB - The present case report illustrates the clinical applicability of the proximal isovelocity surface area (PISA) method in identifying, locating and assessing paravalvular prosthetic mitral regurgitation by transthoracic echocardiography. PMID- 19216753 TI - Bovine renal lipofuscinosis: prevalence, genetics and impact on milk production and weight at slaughter in Danish cattle. AB - BACKGROUND: Bovine renal lipofuscinosis (BRL) is an incidental finding in cattle at slaughter. Condemnation of the kidneys as unfit for human consumption was until recently considered the only implication of BRL. Recent studies have indicated a negative influence on the health of affected animals. The present study investigated the prevalence, genetics and effect of BRL on milk yield and weight at slaughter. METHODS: BRL status of slaughter cattle was recorded at four abattoirs during a 2-year-period. Data regarding breed, age, genetic descent, milk yield and weight at slaughter were extracted from the Danish Cattle Database. The prevalence of BRL was estimated stratified by breed and age-group. Furthermore, total milk yield, milk yield in last full lactation and weight at slaughter were compared for BRL-affected and non-affected Danish Holsteins and Danish Red cattle. RESULTS: 433,759 bovines were slaughtered and 787 of these had BRL. BRL was mainly diagnosed in Danish Red, Danish Holstein and crossbreds. The age of BRL affected animals varied from 11 months to 13 years, but BRL was rarely diagnosed in cattle less than 2 years of age.The total lifelong energy corrected milk (ECM) yields were 3,136 and 4,083 kg higher for BRL affected Danish Red and Danish Holsteins, respectively. However, the median life span of affected animals was 4.9 months longer, and age-corrected total milk yield was 1,284 kg lower for BRL affected Danish Red cows. These cows produced 318 kg ECM less in their last full lactation. Weight at slaughter was not affected by BRL status.The cases occurred in patterns consistent with autosomal recessive inheritance and several family clusters of BRL were found. Analysis of segregation ratios demonstrated the expected ratio for Danish Red cattle, but not for Danish Holsteins. CONCLUSION: The study confirmed that BRL is a common finding in Danish Holsteins and Danish Red cattle at slaughter. The disorder is associated with increased total milk yield due to a longer production life. However, a reduced milk yield was detected in the end of the production life in Danish Red. The study supports that BRL is inherited autosomal recessively in the Danish Red breed and Danish Holsteins, but with incomplete penetrance of the genotype in Danish Holsteins. PMID- 19216752 TI - A single immunization with HA DNA vaccine by electroporation induces early protection against H5N1 avian influenza virus challenge in mice. AB - BACKGROUND: Developing vaccines for the prevention of human infection by H5N1 influenza viruses is an urgent task. DNA vaccines are a novel alternative to conventional vaccines and should contribute to the prophylaxis of emerging H5N1 virus. In this study, we assessed whether a single immunization with plasmid DNA expressing H5N1 hemagglutinin (HA) could provide early protection against lethal challenge in a mouse model. METHODS: Mice were immunized once with HA DNA at 3, 5, 7 days before a lethal challenge. The survival rate, virus titer in the lungs and change of body weight were assayed to evaluate the protective abilities of the vaccine. To test the humoral immune response induced by HA DNA, serum samples were collected through the eye canthus of mice on various days after immunization and examined for specific antibodies by ELISA and an HI assay. Splenocytes were isolated after the immunization to determine the antigen-specific T-cell response by the ELISPOT assay. RESULTS: Challenge experiments revealed that a single immunization of H5N1 virus HA DNA is effective in early protection against lethal homologous virus. Immunological analysis showed that an antigen-specific antibody and T-cell response could be elicited in mice shortly after the immunization. The protective abilities were correlated with the amount of injected DNA and the length of time after vaccination. CONCLUSION: A single immunization of 100 mug H5 HA DNA vaccine combined with electroporation was able to provide early protection in mice against homologous virus infection. PMID- 19216754 TI - Knowledge about childhood autism and opinion among healthcare workers on availability of facilities and law caring for the needs and rights of children with childhood autism and other developmental disorders in Nigeria. AB - BACKGROUND: In designing programs to raise the community level of awareness about childhood autism in sub-Saharan Africa, it is logical to use the primary healthcare workers as contact point for education of the general public. Tertiary healthcare workers could play the role of trainers on childhood autism at primary healthcare level. Assessing their baseline knowledge about childhood autism to detect areas of knowledge gap is an essential ingredient in starting off such programs that would be aimed at early diagnosis and interventions. Knowledge of the healthcare workers on availability of facilities and law that would promote the required interventions is also important. This study assessed the baseline knowledge about childhood autism and opinion among Nigerian healthcare workers on availability of facilities and law caring for the needs and rights of children with childhood autism and other developmental disorders. METHOD: A total of one hundred and thirty four (134) consented healthcare workers working in tertiary healthcare facilities located in south east and south-south regions of Nigeria were interviewed with Socio-demographic, Knowledge about Childhood Autism among Health Workers (KCAHW) and Opinion on availability of Facilities and Law caring for the needs and rights of children with Childhood Autism and other developmental disorders (OFLCA) questionnaires. RESULTS: The total mean score of participated healthcare workers on KCAHW questionnaire was 12.35 +/- 4.40 out of a total score of 19 possible. Knowledge gap was found to be higher in domain 3 (symptoms of obsessive and repetitive pattern of behavior), followed by domains 1 (symptoms of impairments in social interaction), 4 (type of disorder autism is and associated co-morbidity) and 2 (symptoms of communication impairments) of KCAHW respectively among the healthcare workers. Knowledge about childhood autism (KCA) as measured by scores on KCAHW questionnaire was significantly associated with age group distribution of the healthcare workers, with those age group of fourth decades and above more likely to have higher mean score (p = 0.004) and previous experience of managing children with autism spectrum disorders (ASD) (p = 0.000). KCA showed near significant association with area of specialty, with those healthcare workers in psychiatry compared to pediatrics having higher mean score (p = 0.071) and also with years of working experience of the healthcare workers (p = 0.056). More than half of the healthcare workers subscribed to the opinion that facilities and law caring for the needs and rights of children with childhood autism and other developmental disorders are lacking in Nigeria. CONCLUSION: The correlates of KCA may help in selection of those tertiary healthcare workers that would best fit the role of trainers. It is important to update the knowledge gaps of those healthcare workers who scored low in different domains of KCAHW questionnaire. It is imperative for policy makers in Nigeria to advocate and implement multidisciplinary healthcare service system that would ensure early diagnosis and interventions. Nationally representative baseline epidemiological data that would guide policy and planning are also desirable. PMID- 19216755 TI - Chmp 1A is a mediator of the anti-proliferative effects of all-trans retinoic acid in human pancreatic cancer cells. AB - BACKGROUND: We recently have shown that Charged multivesicular protein/Chromatin modifying protein1A (Chmp1A) functions as a tumor suppressor in human pancreatic tumor cells. Pancreatic cancer has the worst prognosis of all cancers with a dismal 5-year survival rate. Preclinical studies using ATRA for treating human pancreatic cancer suggest this compound might be useful for treatment of pancreatic cancer patients. However, the molecular mechanism by which ATRA inhibits growth of pancreatic cancer cells is not clear. The objective of our study was to investigate whether Chmp1A is involved in ATRA-mediated growth inhibition of human pancreatic tumor cells. RESULTS: We performed microarray studies using HEK 293T cells and discovered that Chmp1A positively regulated Cellular retinol-binding protein 1 (CRBP-1). CRBP-1 is a key regulator of All trans retinoic acid (ATRA) through ATRA metabolism and nuclear localization. Since our microarray data indicates a potential involvement of Chmp1A in ATRA signaling, we tested this hypothesis by treating pancreatic tumor cells with ATRA in vitro. In the ATRA-responsive cell lines, ATRA significantly increased the protein expression of Chmp1A, CRBP-1, P53 and phospho-P53 at serine 15 and 37 position. We found that knockdown of Chmp1A via shRNA abolished the ATRA-mediated growth inhibition of PanC-1 cells. Also, Chmp1A silencing diminished the increase of Chmp1A, P53 and phospho-P53 protein expression induced by ATRA. In the ATRA non-responsive cells, ATRA did not have any effect on the protein level of Chmp1A and P53. Chmp1A over-expression, however, induced growth inhibition of ATRA non responsive cells, which was accompanied by an increase of Chmp1A, P53 and phospho P53. Interestingly, in ATRA responsive cells Chmp1A is localized to the nucleus, which became robust upon ATRA treatment. In the ATRA-non-responsive cells, Chmp1A was mainly translocated to the plasma membrane upon ATRA treatment. CONCLUSION: Collectively our data provides evidence that Chmp1A mediates the growth inhibitory activity of ATRA in human pancreatic cancer cells via regulation of CRBP-1. Our results also suggest that nuclear localization of Chmp1A is important in mediating ATRA signaling. PMID- 19216756 TI - Interstitial pneumonitis following intrapleural chemotherapy. AB - BACKGROUND: Mucinous neoplasms within the abdomen may disseminate by direct extension through the diaphragm to involve the pleural space. Treatment of this condition is by parietal and visceral pleurectomy followed by hyperthermic intrapleural chemotherapy. CASE PRESENTATION: In this case report a patient developed persistent right upper lobe interstitial pneumonitis and progressive parenchymal fibrosis following intrapleural chemotherapy treatment with mitomycin C and doxrubicin. The condition persisted until death 28 months later. Death was from progressive intraabdominal disease with intestinal obstruction and sepsis associated with progressive pulmonary parenchymal disease. The right pleural space disease did not recur. CONCLUSION: This manuscript is the first case report describing interstitial pneumonitis and lung fibrosis following intrapleural chemotherapy. Since pulmonary toxicity from chemotherapy is a dose-dependent phenomenon, dose reduction of intrapleural as compared to intraperitoneal hyperthermic chemotherapy may be necessary. PMID- 19216757 TI - Massively parallel pyrosequencing highlights minority variants in the HIV-1 env quasispecies deriving from lymphomonocyte sub-populations. AB - BACKGROUND: Virus-associated cell membrane proteins acquired by HIV-1 during budding may give information on the cellular source of circulating virions. In the present study, by applying immunosorting of the virus and of the cells with antibodies targeting monocyte (CD36) and lymphocyte (CD26) markers, it was possible to directly compare HIV-1 quasispecies archived in circulating monocytes and T lymphocytes with that present in plasma virions originated from the same cell types. Five chronically HIV-1 infected patients who underwent therapy interruption after prolonged HAART were enrolled in the study. The analysis was performed by the powerful technology of ultra-deep pyrosequencing after PCR amplification of part of the env gene, coding for the viral glycoprotein (gp) 120, encompassing the tropism-related V3 loop region. V3 amino acid sequences were used to establish heterogeneity parameters, to build phylogenetic trees and to predict co-receptor usage. RESULTS: The heterogeneity of proviral and viral genomes derived from monocytes was higher than that of T-lymphocyte origin. Both monocytes and T lymphocytes might contribute to virus rebounding in the circulation after therapy interruptions, but other virus sources might also be involved. In addition, both proviral and circulating viral sequences from monocytes and T lymphocytes were predictive of a predominant R5 coreceptor usage. However, minor variants, segregating from the most frequent quasispecies variants, were present. In particular, in proviral genomes harboured by monocytes, minority variant clusters with a predicted X4 phenotype were found. CONCLUSION: This study provided the first direct comparison between the HIV-1 quasispecies archived as provirus in circulating monocytes and T lymphocytes with that of plasma virions replicating in the same cell types. Ultra-deep pyrosequencing generated data with some order of magnitude higher than any previously obtained with conventional approaches. Next generation sequencing allowed the analysis of previously inaccessible aspects of HIV-1 quasispecies, such as co-receptor usage of minority variants present in archived proviral sequences and in actually replicating virions, which may have clinical and therapeutic relevance. PMID- 19216758 TI - AICAR activates the pluripotency transcriptional network in embryonic stem cells and induces KLF4 and KLF2 expression in fibroblasts. AB - BACKGROUND: Pluripotency, the property of a cell to differentiate into all cellular types of a given organism, is central to the development of stem cell based therapies and regenerative medicine. Stem cell pluripotency is the result of the orchestrated activation of a complex transcriptional network characterized by the expression of a set of transcription factors including the master regulators of pluripotency Nanog and Oct4. Recently, it has been shown that pluripotency can be induced in somatic cells by viral-mediated expression of the transcription factors Oct3/4, Sox2, Klf4, and c-Myc. RESULTS: Here we show that 5 Aminoimidazole-4-carboxamide-1-b-riboside (AICAR) is able to activate the molecular circuitry of pluripotency in mouse embryonic stem cells (mESC) and maintain Nanog and Oct4 expression in mESC exposed to the differentiating agent retinoic acid. We also show that AICAR is able to induce Klf4, Klf2 and Myc expression in both mESC and murine fibroblasts. CONCLUSION: AICAR is able to activate the molecular circuitry of pluripotency in mESC and to induce the expression of several key regulators of pluripotency in somatic cells. AICAR is therefore a useful pharmacological entity for studying small molecule mediated induction of pluripotency. PMID- 19216759 TI - Rural Gambian women's reliance on health workers to deliver sulphadoxine pyrimethamine as recommended intermittent preventive treatment for malaria in pregnancy. AB - BACKGROUND: The use of most anti-malarial medications is restricted during pregnancy, but two doses of sulphadoxine-pyrimethamine are recommended after the first trimester as intermittent preventive treatment in pregnancy (IPTp). In The Gambia, only 32% of women receive two doses and very little research has been conducted on women's awareness of drug safety during pregnancy. The objective of this paper was to assess whether rural Gambian women were aware of the importance of the timing of the two-dose IPT dose schedule and its relevance to drug safety. METHODS: This was a qualitative study in which 41 interviews and 16 focus group discussions with women, adolescents, men and traditional birth attendants were conducted. A generic qualitative approach was used to generate a theory as to why women might not participate in IPTp as recommended. RESULTS: Although most women used calendar months to count their stage of pregnancy, these months did not correlate with their concept of foetal development. Foetal growth was described following Islamic tradition as water, clot, piece of meat and human being, although there was little consensus about the order or timing in which these stages occurred. Common signs and conditions of malaria were known. Women were anxious about miscarriage and recognized that some medicines should not be taken in the first trimester, but were urged by men and traditional birth attendants to attend for antenatal care in the first trimester to "start treatment." General knowledge about the purpose of pregnancy medications and when they should be taken was poor among both men and women. One important result was that women relied entirely on health workers to provide safe drugs, at the correct time. CONCLUSION: Women did not have relevant information to judge the safety and appropriate timing of pregnancy drugs, which made them over-reliant on health workers. They should be encouraged to date their own pregnancies in culturally relevant terms and to anticipate when and which medications they should receive. PMID- 19216760 TI - Search for cardiac calcium cycling gene mutations in familial ventricular arrhythmias resembling catecholaminergic polymorphic ventricular tachycardia. AB - BACKGROUND: Catecholaminergic polymorphic ventricular tachycardia (CPVT) is a severe inherited cardiac disorder caused by mutations predominantly in the ryanodine receptor (RyR2) gene. We sought to identify mutations in genes affecting cardiac calcium cycling in patients with CPVT and in less typical familial exercise-related ventricular arrhythmias. METHODS AND RESULTS: We recruited 33 consecutive patients with frequent ventricular premature complexes (VPCs) without structural heart disease and often history of syncope or sudden death in family. Sixteen of the patients featured a phenotype typical of CPVT. In 17 patients, VPCs emerged also at rest. Exercise stress test and echocardiography were performed to each patient and 232 family members. Familial background was evident in 42% of cases (n = 14). We sequenced all the coding exons of the RyR2, FKBP1B, ATP2A2 and SLC8A1 genes from the index patients. Single channel recordings of a mutant RyR2 were performed in planar lipid bilayers. Two novel RyR2 missense mutations (R1051P and S616L) and two RyR2 exon 3 deletions were identified, explaining 25% of the CPVT phenotypes. A rare variant (N3308S) with open probabilities similar to the wild type channels in vitro, was evident in a patient with resting VPCs. No disease-causing variants were detectable in the FKBP1B, ATP2A2 or SLC8A1 genes. CONCLUSION: We report two novel CPVT-causing RyR2 mutations and a novel RyR2 variant of uncertain clinical significance in a patient with abundant resting VPCs. Our data also strengthen the previous assumption that exon 3 deletions of RyR2 should screened for in CPVT and related phenotypes. PMID- 19216761 TI - Lbx2 regulates formation of myofibrils. AB - BACKGROUND: Skeletal muscle differentiation requires assembly of contractile proteins into organized myofibrils. The Drosophila ladybird homeobox gene (lad) functions in founder cells of the segmental border muscle to promote myoblast fusion and muscle shaping. Tetrapods have two homologous genes (Lbx). Lbx1 functions in migration and/or proliferation of hypaxial myoblasts, whereas the function of Lbx2 is poorly understood. RESULTS: To elucidate the role of Lbx in vertebrate myogenesis, we examined Lbx function in zebrafish. Zebrafish lbx2 transcripts appear in newly formed paraxial mesoderm and become restricted to adaxial cells, precursors of slow muscle. Slow muscles lose lbx2 expression as they differentiate, while a subset of differentiating fast muscle cells transiently expresses lbx2. Fin and hyoid muscle express lbx2 later. In contrast, lbx1b expression first appears lateral to the somites at late segmentation stages and is later restricted to fin muscle. Morpholino knockdown of Lbx1b and Lbx2 suppresses hypaxial muscle development. Moreover, knockdown of Lbx2 results in malformation of muscle fibers and reduced fusion of fast precursors, although no obvious effects on induction or specification are observed. Expression of myofilament genes, including actin and myosin, requires the engrailed repressor domain of Lbx2. CONCLUSION: Our results elucidate a new function of Lbx2 as a regulator of myofibril formation. PMID- 19216762 TI - Process evaluation of a community-based program for prevention and control of non communicable disease in a developing country: The Isfahan Healthy Heart Program, Iran. AB - BACKGROUND: Cardiovascular diseases are the most common cause of mortality in Iran. A six-year, comprehensive, integrated community-based demonstration study entitled Isfahan Healthy Heart Program (IHHP) conducted in Iran, and it started in 2000. Evaluation and monitoring are integrated parts of this quasi experimental trial, and consists of process, as well as short and long-term impact evaluations. This paper presents the design of the "process evaluation" for IHHP, and the results pertaining to some interventional strategies that were implemented in workplaces METHODS: The process evaluation addresses the internal validity of IHHP by ascertaining the degree to which the program was implemented as intended. The IHHP process evaluation is a triangulated study conducted for all interventions at their respective venues. All interventional activities are monitored to determine why and how some are successful and sustainable, to identify mechanisms as well as barriers and facilitators of implementation. RESULTS: The results suggest that factory workers and managers are satisfied with the interventions. In the current study, success was mainly shaped by the organizational readiness and timing of the implementation. Integrating most of activities of the project to the existing ongoing activities of public health officers in worksites is suggested to be the most effective means of implementation of the health promoting activities in workplaces. CONCLUSION: The results of our experience may help other developing countries to plan for similar interventions. PMID- 19216763 TI - Effectiveness of manual therapy compared to usual care by the general practitioner for chronic tension-type headache: design of a randomised clinical trial. AB - BACKGROUND: Patients with Chronic Tension Type Headache (CTTH) report functional and emotional impairments (loss of workdays, sleep disturbances, emotional well being) and are at risk for overuse of medication. Manual therapy may improve symptoms through mobilisation of the spine, correction of posture, and training of cervical muscles.We present the design of a randomised clinical trial (RCT) evaluating the effectiveness of manual therapy (MT) compared to usual care by the general practitioner (GP) in patients with CTTH. METHODS AND DESIGN: Patients are eligible for participation if they present in general practice with CTTH according to the classification of the International Headache Society (IHS).Participants are randomised to either usual GP care according to the national Dutch general practice guidelines for headache, or manual therapy, consisting of mobilisations (high- and low velocity techniques), exercise therapy for the cervical and thoracic spine and postural correction. The primary outcome measures are the number of headache days and use of medication. Secondary outcome measures are severity of headache, functional status, sickness absence, use of other healthcare resources, active cervical range of motion, algometry, endurance of the neckflexor muscles and head posture. Follow-up assessments are conducted after 8 and 26 weeks. DISCUSSION: This is a pragmatic trial in which interventions are offered as they are carried out in everyday practice. This increases generalisability of results, but blinding of patients, GPs and therapists is not possible.The results of this trial will contribute to clinical decision making of the GP regarding referral to manual therapy in patients with chronic tension headache. PMID- 19216764 TI - PDGF/VEGF signaling controls cell size in Drosophila. AB - BACKGROUND: In multicellular animals, cell size is controlled by a limited set of conserved intracellular signaling pathways, which when deregulated contribute to tumorigenesis by enabling cells to grow outside their usual niche. To delineate the pathways controlling this process, we screened a genome-scale, image-based Drosophila RNA interference dataset for double-stranded RNAs that reduce the average size of adherent S2R+ cells. RESULTS: Automated analysis of images from this RNA interference screen identified the receptor tyrosine kinase Pvr, Ras pathway components and several novel genes as regulators of cell size. Significantly, Pvr/Ras signaling also affected the size of other Drosophila cell lines and of larval hemocytes. A detailed genetic analysis of this growth signaling pathway revealed a role for redundant secreted ligands, Pvf2 and Pvf3, in the establishment of an autocrine growth signaling loop. Downstream of Ras1, growth signaling was found to depend on parallel mitogen-activated protein kinase (MAPK) and phospho-inositide-3-kinase (PI3K) signaling modules, as well as the Tor pathway. CONCLUSIONS: This automated genome-wide screen identifies autocrine Pvf/Pvr signaling, upstream of Ras, MAPK and PI3K, as rate-limiting for the growth of immortalized fly cells in culture. Since, Pvf2/3 and Pvr show mutually exclusive in vivo patterns of gene expression, these data suggest that co expression of this receptor-ligand pair plays a key role in driving cell autonomous growth during the establishment of Drosophila cell lines, as has been suggested to occur during tumor development. PMID- 19216765 TI - Anti-malarial activities of Andrographis paniculata and Hedyotis corymbosa extracts and their combination with curcumin. AB - BACKGROUND: Herbal extracts of Andrographis paniculata (AP) and Hedyotis corymbosa (HC) are known as hepato-protective and fever-reducing drugs since ancient time and they have been used regularly by the people in the south Asian sub-continent. Methanolic extracts of these two plants were tested in vitro on choloroquine sensitive (MRC-pf-20) and resistant (MRC-pf-303) strains of Plasmodium falciparum for their anti-malarial activity. METHODS: Growth inhibition was determined using different concentrations of these plant extracts on synchronized P. falciparum cultures at the ring stage. The interactions between these two plant extracts and individually with curcumin were studied in vitro. The performance of these two herbal extracts in isolation and combination were further evaluated in vivo on Balb/c mice infected with Plasmodium berghei ANKA and their efficacy was compared with that of curcumin. The in vivo toxicity of the plant derived compounds as well as their parasite stage-specificity was studied. RESULTS: The 50% inhibitory concentration (IC50) of AP (7.2 microg/ml) was found better than HC (10.8 microg/ml). Combination of these two herbal drugs showed substantial enhancement in their anti-malarial activity. Combinatorial effect of each of these with curcumin also revealed anti-malarial effect. Additive interaction between the plant extracts (AP + HC) and their individual synergism with curcumin (AP+CUR, HC+CUR) were evident from this study. Increased in vivo potency was also observed with the combination of plant extracts over the individual extracts and curcumin. Both the plant extracts were found to inhibit the ring stage of the parasite and did not show any in vivo toxicity, whether used in isolation or in combination. CONCLUSION: Both these two plant extracts in combination with curcumin could be an effective, alternative source of herbal anti-malarial drugs. PMID- 19216766 TI - Specialist training in Fiji: why do graduates migrate, and why do they remain? A qualitative study. AB - BACKGROUND: Specialist training was established in the late 1990s at the Fiji School of Medicine. Losses of graduates to overseas migration and to the local private sector prompted us to explore the reasons for these losses from the Fiji public workforce. METHODS: Data were collected on the whereabouts and highest educational attainments of the 66 Fiji doctors who had undertaken specialist training to at least the diploma level between 1997 and 2004. Semistructured interviews focusing on career decisions were carried out with 36 of these doctors, who were purposely sampled to include overseas migrants, temporary overseas trainees, local private practitioners and public sector doctors. RESULTS: 120 doctors undertook specialist training to at least the diploma level between 1997 and 2004; 66 of the graduates were Fiji citizens or permanent residents; 54 originated from other countries in the region. Among Fiji graduates, 42 completed a diploma and 24 had either completed (21) or were enrolled (3) in a master's programme. Thirty-two (48.5%) were working in the public sectors, four (6.0%) were temporarily training overseas, 30.3% had migrated overseas and the remainder were mostly in local private practice. Indo Fijian ethnicity and non-completion of full specialist training were associated with lower retention in the public sectors, while gender had little impact. Decisions to leave the public sectors were complex, with concerns about political instability and family welfare predominating for overseas migrants, while working conditions not conducive to family life or frustrations with career progression predominated for local private practitioners. Doctors remaining in the public sectors reported many satisfying aspects to their work despite frustrations, though 40% had seriously considered resigning from the public service and 60% were unhappy with their career progression. CONCLUSION: Overall, this study provides some support for the view that local or regional postgraduate training may increase retention of doctors. Attention to career pathways and other sources of frustration, in addition to encouragement to complete training, should increase the likelihood of such programmes' reaching their full potentials. PMID- 19216767 TI - The extent of population genetic subdivision differs among four co-distributed shark species in the Indo-Australian archipelago. AB - BACKGROUND: The territorial fishing zones of Australia and Indonesia are contiguous to the north of Australia in the Timor and Arafura Seas and in the Indian Ocean to the north of Christmas Island. The area surrounding the shared boundary consists of a variety of bio-diverse marine habitats including shallow continental shelf waters, oceanic trenches and numerous offshore islands. Both countries exploit a variety of fisheries species, including whaler (Carcharhinus spp.) and hammerhead sharks (Sphyrna spp.). Despite their differences in social and financial arrangements, the two countries are motivated to develop complementary co-management practices to achieve resource sustainability. An essential starting point is knowledge of the degree of population subdivision, and hence fisheries stock status, in exploited species. RESULTS: Populations of four commercially harvested shark species (Carcharhinus obscurus, Carcharhinus sorrah, Prionace glauca, Sphyrna lewini) were sampled from northern Australia and central Indonesia. Neutral genetic markers (mitochondrial DNA control region sequence and allelic variation at co-dominant microsatellite loci) revealed genetic subdivision between Australian and Indonesian populations of C. sorrah. Further research is needed to address the possibility of genetic subdivision among C. obscurus populations. There was no evidence of genetic subdivision for P. glauca and S. lewini populations, but the sampling represented a relatively small part of their distributional range. For these species, more detailed analyses of population genetic structure is recommended in the future. CONCLUSION: Cooperative management between Australia and Indonesia is the best option at present for P. glauca and S. lewini, while C. sorrah and C. obscurus should be managed independently. On-going research on these and other exploited shark and ray species is strongly recommended. Biological and ecological similarity between species may not be a predictor of population genetic structure, so species-specific studies are recommended to provide new data to assist with sustainable fisheries management. PMID- 19216768 TI - Evaluating the association of common APOA2 variants with type 2 diabetes. AB - BACKGROUND: APOA2 is a positional and biological candidate gene for type 2 diabetes at the chromosome 1q21-q24 susceptibility locus. The aim of this study was to examine if HapMap phase II tag SNPs in APOA2 are associated with type 2 diabetes and quantitative traits in French Caucasian subjects. METHODS: We genotyped the three HapMap phase II tagging SNPs (rs6413453, rs5085 and rs5082) required to capture the common variation spanning the APOA2 locus in our type 2 diabetes case-control cohort comprising 3,093 French Caucasian subjects. The association between these variants and quantitative traits was also examined in the normoglycaemic adults of the control cohort. In addition, meta-analysis of publicly available whole genome association data was performed. RESULTS: None of the APOA2 tag SNPs were associated with type 2 diabetes in the French Caucasian case-control cohort (rs6413453, P = 0.619; rs5085, P = 0.245; rs5082, P = 0.591). However, rs5082 was marginally associated with total cholesterol levels (P = 0.026) and waist-to-hip ratio (P = 0.029). The meta-analysis of data from 12,387 subjects confirmed our finding that common variation at the APOA2 locus is not associated with type 2 diabetes. CONCLUSION: The available data does not support a role for common variants in APOA2 on type 2 diabetes susceptibility or related quantitative traits in Northern Europeans. PMID- 19216769 TI - Rapid glycation with D-ribose induces globular amyloid-like aggregations of BSA with high cytotoxicity to SH-SY5Y cells. AB - BACKGROUND: D-ribose in cells and human serum participates in glycation of proteins resulting in advanced glycation end products (AGEs) that affect cell metabolism and induce cell death. However, the mechanism by which D-ribose glycated proteins induce cell death is still unclear. RESULTS: Here, we incubated D-ribose with bovine serum albumin (BSA) and observed changes in the intensity of fluorescence at 410 nm and 425 nm to monitor the formation of D-ribose-glycated BSA. Comparing glycation of BSA with xylose (a control for furanose), glucose and fructose (controls for pyranose), the rate of glycation with D-ribose was the most rapid. Protein intrinsic fluorescence (335 nm), Nitroblue tetrazolium (NBT) assays and Western blotting with anti-AGEs showed that glycation of BSA incubated with D-ribose occurred faster than for the other reducing sugars. Protein intrinsic fluorescence showed marked conformational changes when BSA was incubated with D-ribose. Importantly, observations with atomic force microscopy showed that D-ribose-glycated BSA appeared in globular polymers. Furthermore, a fluorescent assay with Thioflavin T (ThT) showed a remarkable increase in fluorescence at 485 nm in the presence of D-ribose-glycated BSA. However, ThT fluorescence did not show the same marked increase in the presence of xylose or glucose. This suggests that glycation with D-ribose induced BSA to aggregate into globular amyloid-like deposits. As observed by Hoechst 33258 staining, 3-(4, 5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and cell counting kit-8 (CCK-8) assay, lactate dehydrogenase (LDH) activity assay, flow cytometry using Annexin V and Propidium Iodide staining and reactive oxygen species (ROS) measurements, the amyloid-like aggregation of glycated BSA induced apoptosis in the neurotypic cell line SH-SY5Y. CONCLUSION: Glycation with D-ribose induces BSA to misfold rapidly and form globular amyloid-like aggregations which play an important role in cytotoxicity to neural cells. PMID- 19216770 TI - Is peer review useful in assessing research proposals in Indigenous health? A case study. AB - BACKGROUND: There has been considerable examination and critique of traditional (academic) peer review processes in quality assessment of grant applications. At the same time, the use of traditional research processes in Indigenous research has been questioned. Many grant funding organisations have changed the composition of their peer review panels to reflect these concerns but the question remains do these reforms go far enough? In this project we asked people working in areas associated with Aboriginal health research in a number of capacities, their views on the use of peer review in assessing Indigenous research proposals. METHODS: In semi-structured interviews we asked 18 individuals associated with an Australian Indigenous research funding organisation to reflect on their experience with peer review in quality assessment of grant applications. We also invited input from a steering group drawn from a variety of organisations involved in Aboriginal research throughout Australia and directly consulted with three Aboriginal-controlled health organisations. RESULTS: There was consensus amongst all participants that traditional academic peer review is inappropriate for quality assessment in Indigenous research. Many expressed the view that using a competitive grant review system in Aboriginal health was counterintuitive, since good research transfer is based on effective collaboration. The consensus within the group favoured a system which built research in a collaborative manner incorporating a variety of different stakeholders in the process. In this system, one-off peer review was still seen as valuable in the form of a "critical friend" who provided advice as to how to improve the research proposal. CONCLUSION: Peer review in the traditional mould should be recognised as inappropriate in Aboriginal research. Building research projects relevant to policy and practice in Indigenous health may require a shift to a new way of selecting, funding and conducting research. PMID- 19216771 TI - Biological behaviors and proteomics analysis of hybrid cell line EAhy926 and its parent cell line A549. AB - BACKGROUND: It is well established that cancer cells can fuse with endothelial cells to form hybrid cells spontaneously, which facilitates cancer cells traversing the endothelial barrier to form metastases. However, up to now, little is known about the biologic characteristics of hybrid cells. Therefore, we investigate the malignant biologic behaviors and proteins expression of the hybrid cell line EAhy926 with its parent cell line A549. METHODS: Cell counting and flow cytometry assay were carried out to assess cell proliferation. The number of cells attached to the extracellular matrix (Matrigel) was measured by MTT assay for the adhesion ability of cells. Transwell chambers were established for detecting the ability of cell migration and invasion. Tumor xenograft test was carried out to observe tumorigenesis of the cell lines. In addition, two dimensional electrophoresis (2-DE) and mass spectrometry were utilized to identify differentially expressed proteins between in Eahy926 cells and in A549 cells. RESULTS: The doubling time of EAhy926 cell and A549 cell proliferation was 25.32 h and 27.29 h, respectively (P > 0.1). Comparing the phase distribution of cell cycle of EAhy926 cells with that of A549 cells, the percentage of cells in G0/G1 phase, in S phase and in G2/M phase was (63.7% +/- 2.65%) VS (60.0% +/- 3.17%), (15.4% +/- 1.52%) VS (13.8% +/- 1.32%), and (20.9% +/- 3.40%) VS (26.3% +/- 3.17%), respectively (P > 0.05). For the ability of cell adhesion of EAhy926 cells and A549 cells, the value of OD in Eahy926 cells was significantly higher than that in A549 cells (0.3236 +/- 0.0514 VS 0.2434 +/- 0.0390, P < 0.004). We also found that the migration ability of Eahy926 cells was stronger than that of A549 cells (28.00 +/- 2.65 VS 18.00 +/- 1.00, P < 0.01), and that the invasion ability of Eahy926 cells was significantly weak than that of A549 cells (15.33 +/ 0.58 VS 26.67 +/- 2.52, P < 0.01). In the xenograft tumor model, expansive masses of classic tumor were found in the A549 cells group, while subcutaneous inflammatory focuses were found in the EAhy926 cells group. Besides, twenty-eight proteins were identified differentially expressed between in EAhy926 cells and in A549 cells by proteomics technologies. CONCLUSION: As for the biological behaviors, the ability of cell proliferation in Eahy926 cells was similar to that in A549 cells, but the ability in adhesion and migration of Eahy926 cells was higher. In addition, Eahy926 cells had weaker ability in invasion and could not form tumor mass. Furthermore, there were many differently expressed proteins between hybrid cell line Eahy926 cells and A549 cells, which might partly account for some of the differences between their biological behaviors at the molecular level. These results may help to understand the processes of tumor angiogenesis, invasion and metastasis, and to search for screening method for more targets for tumor therapy in future. PMID- 19216772 TI - Physician supply forecast: better than peering in a crystal ball? AB - BACKGROUND: Anticipating physician supply to tackle future health challenges is a crucial but complex task for policy planners. A number of forecasting tools are available, but the methods, advantages and shortcomings of such tools are not straightforward and not always well appraised. Therefore this paper had two objectives: to present a typology of existing forecasting approaches and to analyse the methodology-related issues. METHODS: A literature review was carried out in electronic databases Medline-Ovid, Embase and ERIC. Concrete examples of planning experiences in various countries were analysed. RESULTS: Four main forecasting approaches were identified. The supply projection approach defines the necessary inflow to maintain or to reach in the future an arbitrary predefined level of service offer. The demand-based approach estimates the quantity of health care services used by the population in the future to project physician requirements. The needs-based approach involves defining and predicting health care deficits so that they can be addressed by an adequate workforce. Benchmarking health systems with similar populations and health profiles is the last approach. These different methods can be combined to perform a gap analysis. The methodological challenges of such projections are numerous: most often static models are used and their uncertainty is not assessed; valid and comprehensive data to feed into the models are often lacking; and a rapidly evolving environment affects the likelihood of projection scenarios. As a result, the internal and external validity of the projections included in our review appeared limited. CONCLUSION: There is no single accepted approach to forecasting physician requirements. The value of projections lies in their utility in identifying the current and emerging trends to which policy-makers need to respond. A genuine gap analysis, an effective monitoring of key parameters and comprehensive workforce planning are key elements to improving the usefulness of physician supply projections. PMID- 19216773 TI - Prevalence of HBV and HCV among blood donors in Kosovo. AB - Hepatitis is disease of the liver caused by the infectious and non-infectious agents. The aim of study was to analyze the prevalence of HBV and HCV among voluntary blood donors in Kosovo, during 2000-2003. The data from National Center for Blood Transfusion of Kosovo were collected and analyzed through descriptive and comparative epidemiological method of retrospective study. All samples were tested by ELISA test. Out of 70348 samples of the blood donors, 3145 were positive. From overall positive samples, 2939 were HBV positive, 192 HCV positive while 14 samples were positive for both viruses. The HBV prevalence among the blood donors of Kosovo is 4.2%, which range Kosovo to the second zone according to the CDC classification of the geographical spread of the HBV infection. The HCV prevalence among the blood donors in Kosovo is 0.3%. Compared to the other European countries this level of prevalence is relatively low. Age group 30-39 years old was presented with 34.8% of cases. The higher number was among the workers, 842 or 26.8%. Based on the results we can conclude that Kosovo have the similar prevalence for HBV and HCV infections as other South East European countries. PMID- 19216774 TI - The extracellular EXO protein mediates cell expansion in Arabidopsis leaves. AB - BACKGROUND: The EXO (EXORDIUM) gene was identified as a potential mediator of brassinosteroid (BR)-promoted growth. It is part of a gene family with eight members in Arabidopsis. EXO gene expression is under control of BR, and EXO overexpression promotes shoot and root growth. In this study, the consequences of loss of EXO function are described. RESULTS: The exo loss of function mutant showed diminished leaf and root growth and reduced biomass production. Light and scanning electron microscopy analyses revealed that impaired leaf growth is due to reduced cell expansion. Epidermis, palisade, and spongy parenchyma cells were smaller in comparison to the wild-type. The exo mutant showed reduced brassinolide-induced cotyledon and hypocotyl growth. In contrast, exo roots were significantly more sensitive to the inhibitory effect of synthetic brassinolide. Apart from reduced growth, exo did not show severe morphological abnormalities. Gene expression analyses of leaf material identified genes that showed robust EXO dependent expression. Growth-related genes such as WAK1, EXP5, and KCS1, and genes involved in primary and secondary metabolism showed weaker expression in exo than in wild-type plants. However, the vast majority of BR-regulated genes were normally expressed in exo. HA- and GFP-tagged EXO proteins were targeted to the apoplast. CONCLUSION: The EXO gene is essential for cell expansion in leaves. Gene expression patterns and growth assays suggest that EXO mediates BR-induced leaf growth. However, EXO does not control BR-levels or BR-sensitivity in the shoot. EXO presumably is involved in a signalling process which coordinates BR responses with environmental or developmental signals. The hypersensitivity of exo roots to BR suggests that EXO plays a diverse role in the control of BR responses in the root. PMID- 19216775 TI - Comparison of cisplatin sensitivity and the 18F fluoro-2-deoxy 2 glucose uptake with proliferation parameters and gene expression in squamous cell carcinoma cell lines of the head and neck. AB - BACKGROUND: The survival of patients with locally advanced head and neck cancer is still poor, with 5-year survival rates of 24-35%. The identification of prognostic and predictive markers at the molecular and cellular level could make it possible to find new therapeutic targets and provide "taylor made" treatments. Established cell lines of human squamous cell carcinoma (HNSCC) are valuable models for identifying such markers.The aim of this study was to establish and characterize a series of cell lines and to compare the cisplatin sensitivity and 18F fluoro-2 deoxy 2 glucose (18F-FDG) uptake of these cell lines with other cellular characteristics, such as proliferation parameters and TP53 and CCND1 status. METHODS: Explant cultures of fresh tumour tissue were cultivated, and six new permanent cell lines were established from 18 HNSCC cases. Successfully grown cell lines were analysed regarding clinical parameters, histological grade, karyotype, DNA ploidy, and index and S-phase fraction (Spf). The cell lines were further characterized with regard to their uptake of 18F-FDG, their sensitivity to cisplatin, as measured by a viability test (crystal violet), and their TP53 and CCND1 status, by fluorescence in situ hybridization (FISH), polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) with DNA sequencing and, for cyclin D1, by immunohistochemistry. RESULTS: Patients with tumours that could be cultured in vitro had shorter disease-free periods and overall survival time than those whose tumours did not grow in vitro, when analysed with the Kaplan-Meier method and the log-rank test. Their tumours also showed more complex karyotypes than tumours from which cell lines could not be established. No correlation was found between TP53 or CCND1 status and 18F-FDG uptake or cisplatin sensitivity. However, there was an inverse correlation between tumour cell doubling time and 18F-FDG uptake. CONCLUSION: In vitro growth of HNSCC cells seem to be an independent prognostic factor, with cell lines being more readily established from aggressive tumours, a phenomenon more dependent on the molecular genetic characteristics of the tumour cells than on tumour location or TNM status. PMID- 19216776 TI - Comparison of the Airtraq and Truview laryngoscopes to the Macintosh laryngoscope for use by Advanced Paramedics in easy and simulated difficult intubation in manikins. AB - BACKGROUND: Paramedics are frequently required to perform tracheal intubation, a potentially life-saving manoeuvre in severely ill patients, in the prehospital setting. However, direct laryngoscopy is often more difficult in this environment, and failed tracheal intubation constitutes an important cause of morbidity. Novel indirect laryngoscopes, such as the Airtraq and Truview laryngoscopes may reduce this risk. METHODS: We compared the efficacy of these devices to the Macintosh laryngoscope when used by 21 Paramedics proficient in direct laryngoscopy, in a randomized, controlled, manikin study. Following brief didactic instruction with the Airtraq and Truview laryngoscopes, each participant took turns performing laryngoscopy and intubation with each device, in an easy intubation scenario and following placement of a hard cervical collar, in a SimMan manikin. RESULTS: The Airtraq reduced the number of optimization manoeuvres and reduced the potential for dental trauma when compared to the Macintosh, in both the normal and simulated difficult intubation scenarios. In contrast, the Truview increased the duration of intubation attempts, and required a greater number of optimization manoeuvres, compared to both the Macintosh and Airtraq devices. CONCLUSION: The Airtraq laryngoscope performed more favourably than the Macintosh and Truview devices when used by Paramedics in this manikin study. Further studies are required to extend these findings to the clinical setting. PMID- 19216777 TI - Supporting cognition in systems biology analysis: findings on users' processes and design implications. AB - BACKGROUND: Current usability studies of bioinformatics tools suggest that tools for exploratory analysis support some tasks related to finding relationships of interest but not the deep causal insights necessary for formulating plausible and credible hypotheses. To better understand design requirements for gaining these causal insights in systems biology analyses a longitudinal field study of 15 biomedical researchers was conducted. Researchers interacted with the same protein-protein interaction tools to discover possible disease mechanisms for further experimentation. RESULTS: Findings reveal patterns in scientists' exploratory and explanatory analysis and reveal that tools positively supported a number of well-structured query and analysis tasks. But for several of scientists' more complex, higher order ways of knowing and reasoning the tools did not offer adequate support. Results show that for a better fit with scientists' cognition for exploratory analysis systems biology tools need to better match scientists' processes for validating, for making a transition from classification to model-based reasoning, and for engaging in causal mental modelling. CONCLUSION: As the next great frontier in bioinformatics usability, tool designs for exploratory systems biology analysis need to move beyond the successes already achieved in supporting formulaic query and analysis tasks and now reduce current mismatches with several of scientists' higher order analytical practices. The implications of results for tool designs are discussed. PMID- 19216778 TI - A comparison on effects of normalisations in the detection of differentially expressed genes. AB - BACKGROUND: Various normalisation techniques have been developed in the context of microarray analysis to try to correct expression measurements for experimental bias and random fluctuations. Major techniques include: total intensity normalisation; intensity dependent normalisation; and variance stabilising normalisation. The aim of this paper is to discuss the impact of normalisation techniques for two-channel array technology on the process of identification of differentially expressed genes. RESULTS: Through three precise simulation plans, we quantify the impact of normalisations: (a) on the sensitivity and specificity of a specified test statistic for the identification of deregulated genes, (b) on the gene ranking induced by the statistic. CONCLUSION: Although we found a limited difference of sensitivities and specificities for the test after each normalisation, the study highlights a strong impact in terms of gene ranking agreement, resulting in different levels of agreement between competing normalisations. However, we show that the combination of two normalisations, such as glog and lowess, that handle different aspects of microarray data, is able to outperform other individual techniques. PMID- 19216779 TI - Overstating the evidence: double counting in meta-analysis and related problems. AB - BACKGROUND: The problem of missing studies in meta-analysis has received much attention. Less attention has been paid to the more serious problem of double counting of evidence. METHODS: Various problems in overstating the precision of results from meta-analyses are described and illustrated with examples, including papers from leading medical journals. These problems include, but are not limited to, simple double counting of the same studies, double counting of some aspects of the studies, inappropriate imputation of results, and assigning spurious precision to individual studies. RESULTS: Some suggestions are made as to how the quality and reliability of meta-analysis can be improved. It is proposed that the key to quality in meta-analysis lies in the results being transparent and checkable. CONCLUSION: Existing quality check lists for meta-analysis do little to encourage an appropriate attitude to combining evidence and to statistical analysis. Journals and other relevant organisations should encourage authors to make data available and make methods explicit. They should also act promptly to withdraw meta-analyses when mistakes are found. PMID- 19216780 TI - The effect of diabetes mellitus on organ dysfunction with sepsis: an epidemiological study. AB - INTRODUCTION: Diabetes mellitus (DM) is one of the most common chronic co-morbid medical conditions in the USA and is frequently present in patients with sepsis. Previous studies reported that people with DM and severe sepsis are less likely to develop acute lung injury (ALI). We sought to determine whether organ dysfunction differed between people with and without DM and sepsis. METHODS: Using the National Hospital Discharge Survey US, sepsis cases from 1979 to 2003 were integrated with DM prevalence from the Centers for Disease Control and Prevention (CDC) Diabetes Surveillance System. RESULTS: During the study period 930 million acute-care hospitalisations and 14.3 million people with DM were identified. Sepsis occurred in 12.5 million hospitalisations and DM was present in 17% of patients with sepsis. In the population, acute respiratory failure was the most common organ dysfunction (13%) followed by acute renal failure (6%). People with DM were less likely to develop acute respiratory failure (9% vs. 14%, p < 0.05) and more likely to develop acute renal failure (13% vs. 7%, p < 0.05). Of people with DM and sepsis, 27% had a respiratory source of infection compared with 34% in people with no DM (p < 0.05). Among patients with a pulmonary source of sepsis, 16% of those with DM and 23% of those with no DM developed acute respiratory failure (p < 0.05); in non-pulmonary sepsis acute respiratory failure occurred in 6% of people with DM and 10% in those with no DM (p < 0.05). CONCLUSIONS: In sepsis, people with diabetes are less likely to develop acute respiratory failure, irrespective of source of infection. Future studies should determine the relationship of these findings to reduced risk of ALI in people with DM and causative mechanisms. PMID- 19216781 TI - Micro-geographic risk factors for malarial infection. AB - BACKGROUND: Knowledge of geography is integral to the study of insect-borne infectious disease such as malaria. This study was designed to evaluate whether geographic parameters are associated with malarial infection in the East Sepik province of Papua New Guinea (PNG), a remote area where malaria is a major cause of morbidity and mortality. METHODS: A global positioning system (GPS) unit was used at each village to collect elevation, latitude and longitude data. Concurrently, a sketch map of each village was generated and the villages were sub-divided into regions of roughly equal populations. Blood samples were taken from subjects in each region using filter paper collection. The samples were later processed using nested PCR for qualitative determination of malarial infection. The area was mapped using the GPS-information and overlaid with prevalence data. Data tables were examined using traditional chi square statistical techniques. A logistic regression analysis was then used to determine the significance of geographic risk factors including, elevation, distance from administrative centre and village of residence. RESULTS: Three hundred and thirty two samples were included (24% of the total estimated population). Ninety-six were positive, yielding a prevalence of 29%. Chi square testing within each village found a non-random distribution of cases across sub-regions (p < 0.05). Multivariate logistic regression techniques suggested malarial infection changed with elevation (OR = 0.64 per 10 m, p < 0.05) and distance from administrative centre (OR = 1.3 per 100 m, p < 0.05). CONCLUSION: These results suggest that malarial infection is significantly and independently associated with lower elevation and greater distance from administrative centre in a rural area in PNG. This type of analysis can provide information that may be used to target specific areas in developing countries for malaria prevention and treatment. PMID- 19216782 TI - A new 2D-based method for myocardial velocity strain and strain rate quantification in a normal adult and paediatric population: assessment of reference values. AB - BACKGROUND: Recent advances in technology have provided the opportunity for off line analysis of digital video-clips of two-dimensional (2-D) echocardiographic images. Commercially available software that follows the motion of cardiac structures during cardiac cycle computes both regional and global velocity, strain, and strain rate (SR). The present study aims to evaluate the clinical applicability of the software based on the tracking algorithm feature (studied for cardiology purposes) and to derive the reference values for longitudinal and circumferential strain and SR of the left ventricle in a normal population of children and young adults. METHODS: 45 healthy volunteers (30 adults: 19 male, 11 female, mean age 37 +/- 6 years; 15 children: 8 male, 7 female, mean age 8 +/- 2 years) underwent transthoracic echocardiographic examination; 2D cine-loops recordings of apical 4-four 4-chamber (4C) and 2-chamber (2C) views and short axis views were stored for off-line analysis. Computer analyses were performed using specific software relying on the algorithm of optical flow analysis, specifically designed to track the endocardial border, installed on a Windows based computer workstation. Inter and intra-observer variability was assessed. RESULTS: The feasibility of measurements obtained with tissue tracking system was higher in apical view (100% for systolic events; 64% for diastolic events) than in short axis view (70% for systolic events; 52% for diastolic events). Longitudinal systolic velocity decreased from base to apex in all subjects (5.22 +/- 1.01 vs. 1.20 +/- 0.88; p < 0.0001). Longitudinal strain and SR significantly increased from base to apex in all subjects (-12.95 +/- 6.79 vs. -14.87 +/- 6.78; p = 0.002; -0.72 +/- 0.39 vs. -0.94 +/- 0.48, p = 0.0001, respectively). Similarly, circumferential strain and SR increased from base to apex (-21.32 +/- 5.15 vs. -27.02 +/- 5.88, p = 0.002; -1.51 +/- 0.37 vs. -1.95 +/- 0.57, p = 0.003, respectively). Values of global systolic SR, both longitudinal and circumferential, were significantly higher in children than in adults (-1.3 +/- 0.2, vs. -1.11 +/- 0.2, p = 0.006; -1.9 +/- 0.6 vs. -1.6 +/- 0.5, p = 0.0265, respectively). No significant differences in longitudinal and circumferential systolic velocities were identified for any segment when comparing adults with children. CONCLUSION: This 2D based tissue tracking system used for computation is reliable and applicable in adults and children particularly for systolic events. Measured with this technology, we have established reference values for myocardial velocity, Strain and SR for both young adults and children. PMID- 19216783 TI - Achieving professional status: Australian podiatrists' perceptions. AB - BACKGROUND: This paper explores the notion of professional status from the perspective of a sample of Australian podiatrists; how it is experienced, what factors are felt to affect it, and how these are considered to influence professional standing within an evolving healthcare system. Underpinning sociological theory is deployed in order to inform and contextualise the study. METHODS: Data were drawn from a series of in-depth semi-structured interviews (n = 21) and focus groups (n = 9) with podiatrists from across four of Australia's eastern states (Queensland, New South Wales, Victoria and Australian Capital Territory), resulting in a total of 76 participants. Semi-structured interview schedules sought to explore podiatrist perspectives on a range of features related to professional status within podiatry in Australia. RESULTS: Central to the retention and enhancement of status was felt to be the development of specialist roles and the maintenance of control over key task domains. Key distinctions in private and public sector environments, and in rural and urban settings, were noted and found to reflect differing contexts for status development. Marketing was considered important to image enhancement, as was the cache attached to the status of the universities providing graduate education. CONCLUSION: Perceived determinants of professional status broadly matched those identified in the wider sociological literature, most notably credentialism, client status, content and context of work (such as specialisation) and an ideological basis for persuading audiences to acknowledge professional status. In an environment of demographic and workforce change, and the resultant policy demands for healthcare service re-design, enhanced opportunities for specialisation appear evident. Under the current model of professionalism, both role flexibility and uniqueness may prove important. PMID- 19216784 TI - APOBEC3G induces a hypermutation gradient: purifying selection at multiple steps during HIV-1 replication results in levels of G-to-A mutations that are high in DNA, intermediate in cellular viral RNA, and low in virion RNA. AB - BACKGROUND: Naturally occurring Vif variants that are unable to inhibit the host restriction factor APOBEC3G (A3G) have been isolated from infected individuals. A3G can potentially induce G-to-A hypermutation in these viruses, and hypermutation could contribute to genetic variation in HIV-1 populations through recombination between hypermutant and wild-type genomes. Thus, hypermutation could contribute to the generation of immune escape and drug resistant variants, but the genetic contribution of hypermutation to the viral evolutionary potential is poorly understood. In addition, the mechanisms by which these viruses persist in the host despite the presence of A3G remain unknown. RESULTS: To address these questions, we generated a replication-competent HIV-1 Vif mutant in which the A3G binding residues of Vif, Y(40)RHHY(44), were substituted with five alanines. As expected, the mutant was severely defective in an A3G-expressing T cell line and exhibited a significant delay in replication kinetics. Analysis of viral DNA showed the expected high level of G-to-A hypermutation; however, we found substantially reduced levels of G-to-A hypermutation in intracellular viral RNA (cRNA), and the levels of G-to-A mutations in virion RNA (vRNA) were even further reduced. The frequencies of hypermutation in DNA, cRNA, and vRNA were 0.73%, 0.12%, and 0.05% of the nucleotides sequenced, indicating a gradient of hypermutation. Additionally, genomes containing start codon mutations and early termination codons within gag were isolated from the vRNA. CONCLUSION: These results suggest that sublethal levels of hypermutation coupled with purifying selection at multiple steps during the early phase of viral replication lead to the packaging of largely unmutated genomes, providing a mechanism by which mutant Vif variants can persist in infected individuals. The persistence of genomes containing mutated gag genes despite this selection pressure indicates that dual infection and complementation can result in the packaging of hypermutated genomes which, through recombination with wild-type genomes, could increase viral genetic variation and contribute to evolution. PMID- 19216785 TI - Somatic, germline and sex hierarchy regulated gene expression during Drosophila metamorphosis. AB - BACKGROUND: Drosophila melanogaster undergoes a complete metamorphosis, during which time the larval male and female forms transition into sexually dimorphic, reproductive adult forms. To understand this complex morphogenetic process at a molecular-genetic level, whole genome microarray analyses were performed. RESULTS: The temporal gene expression patterns during metamorphosis were determined for all predicted genes, in both somatic and germline tissues of males and females separately. Temporal changes in transcript abundance for genes of known functions were found to correlate with known developmental processes that occur during metamorphosis. We find that large numbers of genes are sex differentially expressed in both male and female germline tissues, and relatively few are sex-differentially expressed in somatic tissues. The majority of genes with somatic, sex-differential expression were found to be expressed in a stage specific manner, suggesting that they mediate discrete developmental events. The Sex-lethal paralog, CG3056, displays somatic, male-biased expression at several time points in metamorphosis. Gene expression downstream of the somatic, sex determination genes transformer and doublesex (dsx) was examined in two-day old pupae, which allowed for the identification of genes regulated as a consequence of the sex determination hierarchy. These include the homeotic gene abdominal A, which is more highly expressed in females as compared to males, as a consequence of dsx. For most genes regulated downstream of dsx during pupal development, the mode of regulation is distinct from that observed for the well-studied direct targets of DSX, Yolk protein 1 and 2. CONCLUSION: The data and analyses presented here provide a comprehensive assessment of gene expression during metamorphosis in each sex, in both somatic and germline tissues. Many of the genes that underlie critical developmental processes during metamorphosis, including sex specific processes, have been identified. These results provide a framework for further functional studies on the regulation of sex-specific development. PMID- 19216786 TI - MODY-like diabetes associated with an apparently balanced translocation: possible involvement of MPP7 gene and cell polarity in the pathogenesis of diabetes. AB - BACKGROUND: Characterization of disease-associated balanced translocations has led to the discovery of genes responsible for many disorders, including syndromes that include various forms of diabetes mellitus. We studied a man with unexplained maturity onset diabetes of the young (MODY)-like diabetes and an apparently balanced translocation [46,XY,t(7;10)(q22;p12)] and sought to identify a novel diabetes locus by characterizing the translocation breakpoints. RESULTS: Mutations in coding exons and splice sites of known MODY genes were first ruled out by PCR amplification and DNA sequencing. Fluorescent in situ hybridization (FISH) studies demonstrated that the translocation did not disrupt two known diabetes-related genes on 10p12. The translocation breakpoints were further mapped to high resolution using FISH and somatic cell hybrids and the junctions PCR-amplified and sequenced. The translocation did not disrupt any annotated transcription unit. However, the chromosome 10 breakpoint was 220 kilobases 5' to the Membrane Protein, Palmitoylated 7 (MPP7) gene, which encodes a protein required for proper cell polarity. This biological function is shared by HNF4A, a known MODY gene. Databases show MPP7 is highly expressed in mouse pancreas and is expressed in human islets. The translocation did not appear to alter lymphoblastoid expression of MPP7 or other genes near the breakpoints. CONCLUSION: The balanced translocation and MODY-like diabetes in the proband could be coincidental. Alternatively, the translocation may cause islet cell dysfunction by altering MPP7 expression in a subtle or tissue-specific fashion. The potential roles of MPP7 mutations in diabetes and perturbed islet cell polarity in insulin secretion warrant further study. PMID- 19216787 TI - The mangled extremity and attempt for limb salvage. AB - BACKGROUND: The decision, whether to amputate or reconstruct a mangled extremity remains the subject of extensive debate since multiple factors influence the decision. METHODS: Sixty three patients with high energy extremity trauma and attempts at limb salvage were retrospectively reviewed. We analyzed 10 cases of massive extremity trauma where there was made an attempt to salvage limbs, although there was a controversy between salvage and amputation. RESULTS: All of the patients except one had major vascular injury and ischemia requiring repair. Three patients died. All of the remaining patients were amputated within 15 days after the salvage procedure, mainly because of extremity sepsis. Seven patients required treatment at the intensive care unit. All patients had at least 2 reconstruction procedures and multiple surgical debridements. CONCLUSION: The functional outcome should be considered realistically before a salvage decision making for extremities with indeterminate prognosis. PMID- 19216788 TI - Pancreatic insulinoma co-existing with gastric GIST in the absence of neurofibromatosis-1. AB - BACKGROUND: Gastrointestinal stromal tumours (GIST) frequently occur in patients with neurofibromatosis type 1 (NF-1). It has been reported that GIST may co-exist with pancreatic endocrine tumors but this has only been in association with NF-1. CASE PRESENTATION: A 76 year old woman presented with a 12 month history of hypoglycaemia symptoms. Abdominal CT scan demonstrated a 13 mm insulinoma localized in the tail of her pancreas. She was commenced on diazoxide and later underwent surgery for enucleation of insulinoma when a small (< 1 cm) incidental tumour was discovered on her stomach wall which was identified as GIST. CONCLUSION: This is the first case report of a pancreatic insulinoma co-existing with a GIST in a patient without NF-1. In addition, we make the first report of rapidly growing cystic GIST recurrence following resection of a primary GIST tumour. PMID- 19216789 TI - The effect of the dual Src/Abl kinase inhibitor AZD0530 on Philadelphia positive leukaemia cell lines. AB - BACKGROUND: Imatinib mesylate, a selective inhibitor of Abl tyrosine kinase, is efficacious in treating chronic myeloid leukaemia (CML) and Ph+ acute lymphoblastic leukaemia (ALL). However, most advanced-phase CML and Ph+ ALL patients relapse on Imatinib therapy. Several mechanisms of refractoriness have been reported, including the activation of the Src-family kinases (SFK). Here, we investigated the biological effect of the new specific dual Src/Abl kinase inhibitor AZD0530 on Ph+ leukaemic cells. METHODS: Cell lines used included BV173 (CML in myeloid blast crisis), SEM t(4;11), Ba/F3 (IL-3 dependent murine pro B), p185Bcr-Abl infected Ba/F3 cells, p185Bcr-Abl mutant infected Ba/F3 cells, SupB15 (Ph+ ALL) and Imatinib resistant SupB15 (RTSupB15) (Ph+ ALL) cells. Cells were exposed to AZD0530 and Imatinib. Cell proliferation, apoptosis, survival and signalling pathways were assessed by dye exclusion, flow cytometry and Western blotting respectively. RESULTS: AZD0530 specifically inhibited the growth of, and induced apoptosis in CML and Ph+ ALL cells in a dose dependent manner, but showed only marginal effects on Ph- ALL cells. Resistance to Imatinib due to the mutation Y253F in p185Bcr-Abl was overcome by AZD0530. Combination of AZD0530 and Imatinib showed an additive inhibitory effect on the proliferation of CML BV173 cells but not on Ph+ ALL SupB15 cells. An ongoing transphosphorylation was demonstrated between SFKs and Bcr-Abl. AZD0530 significantly down-regulated the activation of survival signalling pathways in Ph+ cells, resistant or sensitive to Imatinib, with the exception of the RTSupB15. CONCLUSION: Our results indicate that AZD0530 targets both Src and Bcr-Abl kinase activity and reduces the leukaemic maintenance by Bcr-Abl. PMID- 19216790 TI - Use of high-density tiling microarrays to identify mutations globally and elucidate mechanisms of drug resistance in Plasmodium falciparum. AB - BACKGROUND: The identification of genetic changes that confer drug resistance or other phenotypic changes in pathogens can help optimize treatment strategies, support the development of new therapeutic agents, and provide information about the likely function of genes. Elucidating mechanisms of phenotypic drug resistance can also assist in identifying the mode of action of uncharacterized but potent antimalarial compounds identified in high-throughput chemical screening campaigns against Plasmodium falciparum. RESULTS: Here we show that tiling microarrays can detect de novo a large proportion of the genetic changes that differentiate one genome from another. We show that we detect most single nucleotide polymorphisms or small insertion deletion events and all known copy number variations that distinguish three laboratory isolates using readily accessible methods. We used the approach to discover mutations that occur during the selection process after transfection. We also elucidated a mechanism by which parasites acquire resistance to the antimalarial fosmidomycin, which targets the parasite isoprenoid synthesis pathway. Our microarray-based approach allowed us to attribute in vitro derived fosmidomycin resistance to a copy number variation event in the pfdxr gene, which enables the parasite to overcome fosmidomycin mediated inhibition of isoprenoid biosynthesis. CONCLUSIONS: We show that newly emerged single nucleotide polymorphisms can readily be detected and that malaria parasites can rapidly acquire gene amplifications in response to in vitro drug pressure. The ability to define comprehensively genetic variability in P. falciparum with a single overnight hybridization creates new opportunities to study parasite evolution and improve the treatment and control of malaria. PMID- 19216792 TI - "Will they just pack up and leave?" - attitudes and intended behaviour of hospital health care workers during an influenza pandemic. AB - BACKGROUND: There is a general consensus that another influenza pandemic is inevitable. Although health care workers (HCWs) are essential to the health system response, there are few studies exploring HCW attitudes to pandemic influenza. The aim of this study was to explore HCWs knowledge, attitudes and intended behaviour towards pandemic influenza. METHODS: Cross-sectional investigation of a convenience sample of clinical and non-clinical HCWs from two tertiary-referral teaching hospitals in Sydney, Australia was conducted between June 4 and October 19, 2007. The self-administered questionnaire was distributed to hospital personal from 40 different wards and departments. The main outcome measures were intentions regarding work attendance and quarantine, antiviral use and perceived preparation. RESULTS: Respondents were categorized into four main groups by occupation: Nursing (47.5%), Medical (26.0%), Allied (15.3%) and Ancillary (11.2%). Our study found that most HCWs perceived pandemic influenza to be very serious (80.9%, n = 873) but less than half were able to correctly define it (43.9%, n = 473). Only 24.8% of respondents believed their department to be prepared for a pandemic, but nonetheless most were willing to work during a pandemic if a patient or colleague had influenza. The main determinants of variation in our study were occupational factors, demographics and health beliefs. Non-clinical staff were significantly most likely to be unsure of their intentions (OR 1.43, p < 0.001). Only 42.5% (n = 459) of respondents considered that neuraminidase inhibitor antiviral medications (oseltamivir/zanamivir) would protect them against pandemic influenza, whereas 77.5% (n = 836) believed that vaccination would be of benefit. CONCLUSION: We identified two issues that could undermine the best of pandemic plans - the first, a low level of confidence in antivirals as an effective measure; secondly, that non-clinical workers are an overlooked group whose lack of knowledge and awareness could undermine pandemic plans. Other issues included a high level of confidence in dietary measures to protect against influenza, and a belief among ancillary workers that antibiotics would be protective. All health care worker strategies should include non clinical and ancillary staff to ensure adequate business continuity for hospitals. HCW education, psychosocial support and staff communication could improve knowledge of appropriate pandemic interventions and confidence in antivirals. PMID- 19216791 TI - Centrosome-associated regulators of the G(2)/M checkpoint as targets for cancer therapy. AB - In eukaryotic cells, control mechanisms have developed that restrain cell-cycle transitions in response to stress. These regulatory pathways are termed cell cycle checkpoints. The G(2)/M checkpoint prevents cells from entering mitosis when DNA is damaged in order to afford these cells an opportunity to repair the damaged DNA before propagating genetic defects to the daughter cells. If the damage is irreparable, checkpoint signaling might activate pathways that lead to apoptosis. Since alteration of cell-cycle control is a hallmark of tumorigenesis, cell-cycle regulators represent potential targets for therapy. The centrosome has recently come into focus as a critical cellular organelle that integrates G(2)/M checkpoint control and repairs signals in response to DNA damage. A growing number of G(2)/M checkpoint regulators have been found in the centrosome, suggesting that centrosome has an important role in G(2)/M checkpoint function. In this review, we discuss centrosome-associated regulators of the G(2)/M checkpoint, the dysregulation of this checkpoint in cancer, and potential candidate targets for cancer therapy. PMID- 19216793 TI - Deletions of neuraminidase and resistance to oseltamivir may be a consequence of restricted receptor specificity in recent H3N2 influenza viruses. AB - BACKGROUND: Influenza viruses attach to cells via sialic acid receptors. The viral neuraminidase (NA) is needed to remove sialic acids so that newly budded virions can disperse. Known mechanisms of resistance to NA inhibitors include mutations in the inhibitor binding site, or mutations in the hemagglutinin that reduce avidity for sialic acid and therefore reduce the requirement for NA activity. RESULTS: Influenza H3N2 isolates A/Oklahoma/323/03 (Fujian-like), A/Oklahoma/1992/05 (California-like), and A/Oklahoma/309/06 (Wisconsin-like) lost NA activity on passage in MDCK cells due to internal deletions in the NA-coding RNA segment. The viruses grow efficiently in MDCK cells despite diminished NA activity. The full length NA enzyme activity is sensitive to oseltamivir but replication of A/Oklahoma/323/03 and A/Oklahoma/309/06 in MDCK cells was resistant to this inhibitor, indicating that NA is not essential for replication. There was no change in HA activity or sequence after the NA activity was lost but the three viruses show distinct, quite restricted patterns of receptor specificity by Glycan Array analysis. Extensive predicted secondary structure in RNA segment 6 that codes for NA suggests the deletions are generated by polymerase skipping over base-paired stem regions. In general the NA deletions were not carried into subsequent passages, and we were unable to plaque-purify virus with a deleted NA RNA segment. CONCLUSION: H3N2 viruses from 2003 to the present have reduced requirement for NA when passaged in MDCK cells and are resistant to NA inhibitors, possibly by a novel mechanism of narrow receptor specificity such that virus particles do not self-aggregate. These viruses delete internal regions of the NA RNA during passage and are resistant to oseltamivir. However, deletions are independently generated at each passage, suggesting that virus with a full length NA RNA segment initiates the first round of infection. PMID- 19216794 TI - Induction of anti-tumor immunity by trifunctional antibodies in patients with peritoneal carcinomatosis. AB - Peritoneal carcinomatosis (PC) from epithelial tumors is a fatal diagnosis without efficient treatment. Trifunctional antibodies (trAb) are novel therapeutic approaches leading to a concerted anti-tumor activity resulting in tumor cell destruction. In addition, preclinical data in mouse tumor models demonstrated the induction of long lasting tumor immunity after treatment with trAb. We describe the induction of anti-tumor specific T-lymphocytes after intraperitoneal administration of trAb in patients with PC.9 patients with progressive PC from gastric (n = 6) and ovarian cancer (n = 2), and cancer of unknown primary (n = 1) received 3 escalating doses of trAb after surgery and/or ineffective chemotherapy. The trAb EpCAM x CD3 (10, 20, 40 microg) or HER2/neu x CD3 (10, 40, 80 microg) were applicated by intraperitoneal infusion. Four weeks after the last trAb application, all patients were restimulated by subdermal injection of trAb + autologous PBMC + irradiated autologous tumor cells. Immunological reactivity was tested by analyzing PBMC for specific tumor reactive CD4+/CD8+ T lymphocytes using an IFN-gamma secretion assay.In 5 of 9 patients, tumor reactive CD4+/CD8+ T-lymphocytes increased significantly, indicating specific anti-tumor immunity. A clinical response (stable disease, partial regression) has been observed in 5 of 9 patients, with a mean time to progression of 3.6 months. Follow-up showed a mean survival of 11.8 months (median 8.0 months) after trAb therapy.TrAb are able to induce anti-tumor immunity after intraperitoneal application and restimulation. The induction of long-lasting anti tumor immunity may provide an additional benefit of the intraperitoneal therapy with trAb and should be further elevated in larger clinical trials. PMID- 19216795 TI - Arrested neural and advanced mesenchymal differentiation of glioblastoma cells comparative study with neural progenitors. AB - BACKGROUND: Although features of variable differentiation in glioblastoma cell cultures have been reported, a comparative analysis of differentiation properties of normal neural GFAP positive progenitors, and those shown by glioblastoma cells, has not been performed. METHODS: Following methods were used to compare glioblastoma cells and GFAP+NNP (NHA): exposure to neural differentiation medium, exposure to adipogenic and osteogenic medium, western blot analysis, immunocytochemistry, single cell assay, BrdU incorporation assay. To characterize glioblastoma cells EGFR amplification analysis, LOH/MSI analysis, and P53 nucleotide sequence analysis were performed. RESULTS: In vitro differentiation of cancer cells derived from eight glioblastomas was compared with GFAP-positive normal neural progenitors (GFAP+NNP). Prior to exposure to differentiation medium, both types of cells showed similar multilineage phenotype (CD44+/MAP2+/GFAP+/Vimentin+/Beta III-tubulin+/Fibronectin+) and were positive for SOX-2 and Nestin. In contrast to GFAP+NNP, an efficient differentiation arrest was observed in all cell lines isolated from glioblastomas. Nevertheless, a subpopulation of cells isolated from four glioblastomas differentiated after serum-starvation with varying efficiency into derivatives indistinguishable from the neural derivatives of GFAP+NNP. Moreover, the cells derived from a majority of glioblastomas (7 out of 8), as well as GFAP+NNP, showed features of mesenchymal differentiation when exposed to medium with serum. CONCLUSION: Our results showed that stable co-expression of multilineage markers by glioblastoma cells resulted from differentiation arrest. According to our data up to 95% of glioblastoma cells can present in vitro multilineage phenotype. The mesenchymal differentiation of glioblastoma cells is advanced and similar to mesenchymal differentiation of normal neural progenitors GFAP+NNP. PMID- 19216797 TI - Identification of pancreatic cancer invasion-related proteins by proteomic analysis. AB - BACKGROUND: Markers of pancreatic cancer invasion were investigated in two clonal populations of the cell line, MiaPaCa-2, Clone #3 (high invasion) and Clone #8 (low invasion) using proteomic profiling of an in vitro model of pancreatic cancer. MATERIALS AND METHODS: Using 2D-DIGE followed by MALDI-TOF MS, two clonal sub-populations of the pancreatic cancer cell line, MiaPaCa-2 with high and low invasive capacities were incubated on matrigel 24 hours prior to analysis to stimulate cell-ECM contact and mimic in vivo interaction with the basement membrane. RESULTS: Sixty proteins were identified as being differentially expressed (> 1.2 fold change and p < or = 0.05) between Clone #3 and Clone #8. Proteins found to have higher abundance levels in the highly invasive Clone #3 compared to the low invasive Clone #8 include members of the chaperone activity proteins and cytoskeleton constituents whereas metabolism-associated and catalytic proteins had lower abundance levels. Differential protein expression levels of ALDH1A1, VIM, STIP1 and KRT18 and GAPDH were confirmed by immunoblot. Using RNAi technology, STIP1 knockdown significantly reduced invasion and proliferation of the highly invasive Clone #3. Knockdown of another target, VIM by siRNA in Clone #3 cells also resulted in decreased invasion abilities of Clone #3. Elevated expression of STIP1 was observed in pancreatic tumour tissue compared to normal pancreas, whereas ALDH1A1 stained at lower levels in pancreatic tumours, as detected by immunohistochemistry. CONCLUSION: Identification of targets which play a role in the highly invasive phenotype of pancreatic cancer may help to understand the biological behaviour, the rapid progression of this cancer and may be of importance in the development of new therapeutic strategies for pancreatic cancer. PMID- 19216796 TI - Hands-on time during cardiopulmonary resuscitation is affected by the process of teambuilding: a prospective randomised simulator-based trial. AB - BACKGROUND: Cardiac arrests are handled by teams rather than by individual health care workers. Recent investigations demonstrate that adherence to CPR guidelines can be less than optimal, that deviations from treatment algorithms are associated with lower survival rates, and that deficits in performance are associated with shortcomings in the process of team-building. The aim of this study was to explore and quantify the effects of ad-hoc team-building on the adherence to the algorithms of CPR among two types of physicians that play an important role as first responders during CPR: general practitioners and hospital physicians. METHODS: To unmask team-building this prospective randomised study compared the performance of preformed teams, i.e. teams that had undergone their process of team-building prior to the onset of a cardiac arrest, with that of teams that had to form ad-hoc during the cardiac arrest. 50 teams consisting of three general practitioners each and 50 teams consisting of three hospital physicians each, were randomised to two different versions of a simulated witnessed cardiac arrest: the arrest occurred either in the presence of only one physician while the remaining two physicians were summoned to help ("ad-hoc"), or it occurred in the presence of all three physicians ("preformed"). All scenarios were videotaped and performance was analysed post-hoc by two independent observers. RESULTS: Compared to preformed teams, ad-hoc forming teams had less hands-on time during the first 180 seconds of the arrest (93 +/- 37 vs. 124 +/- 33 sec, P < 0.0001), delayed their first defibrillation (67 +/- 42 vs. 107 +/- 46 sec, P < 0.0001), and made less leadership statements (15 +/- 5 vs. 21 +/- 6, P < 0.0001). CONCLUSION: Hands-on time and time to defibrillation, two performance markers of CPR with a proven relevance for medical outcome, are negatively affected by shortcomings in the process of ad-hoc team-building and particularly deficits in leadership. Team-building has thus to be regarded as an additional task imposed on teams forming ad-hoc during CPR. All physicians should be aware that early structuring of the own team is a prerequisite for timely and effective execution of CPR. PMID- 19216798 TI - Database mining for selection of SNP markers useful in admixture mapping. AB - BACKGROUND: New technologies make it possible for the first time to genotype hundreds of thousands of SNPs simultaneously. A wealth of genomic information in the form of publicly available databases is underutilized as a potential resource for uncovering functionally relevant markers underlying complex human traits. Given the huge amount of SNP data available from the annotation of human genetic variation, data mining is a reasonable approach to investigating the number of SNPs that are informative for ancestry information. METHODS: The distribution and density of SNPs across the genome of African and European populations were extensively investigated by using the HapMap, Affymetrix, and Illumina SNP databases. We exploited these resources by mining the data available from each of these databases to prioritize potential candidate SNPs useful for admixture mapping in complex human diseases and traits. Over 4 million SNPs were compared between Africans and Europeans on the basis of a pre-specified recommended allele frequency difference (delta) value of >or= 0.3. RESULTS: The method identified 15% of HapMap, 11% of Affymetrix, and 14% of Illumina SNP sets as candidate SNPs, termed ancestry informative markers (AIMs). These AIM panels with assigned rs numbers, allele frequencies in each ethnic group, delta value, and map positions are all posted on our website http://www.ssg.uab.edu/downloads/admixture_mapping/SNPAIMs.txt. All marker information in this data set is freely and publicly available without restriction. CONCLUSION: The selected SNP sets represent valuable resources for admixture mapping studies. The overlap between selected AIMs by this single measure of marker informativeness in the different platforms is discussed. PMID- 19216799 TI - Knockdown of astrocyte elevated gene-1 inhibits proliferation and enhancing chemo sensitivity to cisplatin or doxorubicin in neuroblastoma cells. AB - BACKGROUND: Astrocyte elevated gene-1 (AEG-1) was originally characterized as a HIV-1-inducible gene in primary human fetal astrocyte. Recent studies highlight a potential role of AEG-1 in promoting tumor progression and metastasis. The aim of this study was to investigate if AEG-1 serves as a potential therapeutic target of human neuroblastoma. METHODS: We employed RNA interference to reduce AEG-1 expression in human neuroblastoma cell lines and analyzed their phenotypic changes. RESULTS: We found that the knockdown of AEG-1 expression in human neuroblastoma cells significantly inhibited cell proliferation and apoptosis. The specific downregulation induced cell arrest in the G0/G1 phase of cell cycle. In the present study, we also observed a significant enhancement of chemo sensitivity to cisplatin and doxorubicin by knockdown of AEG-1. CONCLUSION: Our study suggests that overexpressed AEG-1 enhance the tumorogenic properties of neuroblastoma cells. The inhibition of AEG-1 expression could be a new adjuvant therapy for neuroblastoma. PMID- 19216800 TI - How are individual-level social capital and poverty associated with health equity? A study from two Chinese cities. AB - BACKGROUND: A growing body of literature has demonstrated that higher social capital is associated with improved health conditions. However, some research indicated that the association between social capital and health was substantially attenuated after adjustment for material deprivation. Studies exploring the association between poverty, social capital and health still have some serious limitations. In China, health equity studies focusing on urban poor are scarce. The purpose of this study is therefore to examine how poverty and individual-level social capital in urban China are associated with health equity. METHODS: Our study is based on a household study sample consisting of 1605 participants in two Chinese cities. For all participants, data on personal characteristics, health status, health care utilisation and social capital were collected. Factor analysis was performed to extract social capital factors. Dichotomised social capital factors were used for logistic regression models. A synergy index (if it is above 1, we can know the existence of the co-operative effect) was computed to examine the interaction effect between lack of social capital and poverty. RESULTS: Results indicated the poor had an obviously higher probability of belonging to the low individual-level social capital group in all the five dimensions, with the adjusted odds ratios ranging from 1.42 to 2.12. When the other variables were controlled for in the total sample, neighbourhood cohesion (NC), and reciprocity and social support (RSS) were statistically associated with poor self-rated health (NC: OR = 1.40; RSS: OR = 1.34). However, for the non-poor sub-sample, no social capital variable was a statistically significant predictor. The synergy index between low individual-level NC and poverty, and between low individual-level RSS and poverty were 1.22 and 1.28, respectively, indicating an aggravating effect between them. CONCLUSION: In this study, we have shown that the interaction effect between poverty and lack of social capital (NC and RSS) was a good predictor of poor SRH in urban China. Improving NC and RSS may be helpful in reducing health inequity; however, poverty reduction is more important and therefore should be implemented at the same time. Policies that attempt to improve health equity via social capital, but neglect poverty intervention, would be counter-productive. PMID- 19216801 TI - The time course of the transcriptomic response of Sinorhizobium meliloti 1021 following a shift to acidic pH. AB - BACKGROUND: The symbiotic soil bacterium Sinorhizobium meliloti often has to face low pH in its natural habitats. To identify genes responding to pH stress a global transcriptional analysis of S. meliloti strain 1021 following a pH shift from pH 7.0 to pH 5.75 was carried out. In detail, oligo-based whole genome microarrays were used in a time course experiment. The monitoring period covered a time span of about one hour after the pH shift. The obtained microarray data was filtered and grouped by K-means clustering in order to obtain groups of genes behaving similarly concerning their expression levels throughout the time course. RESULTS: The results display a versatile response of S. meliloti 1021 represented by distinct expression profiles of subsets of genes with functional relation. The eight generated clusters could be subdivided into a group of four clusters containing genes that were up-regulated and another group of four clusters containing genes that were down-regulated in response to the acidic pH shift. The respective mean expression progression of the four up-regulated clusters could be described as (i) permanently and strong, (ii) permanently and intermediate, (iii) permanently and progressive, and (iv) transiently up-regulated. The expression profile of the four down-regulated clusters could be characterized as (i) permanently, (ii) permanently and progressive, (iii) transiently, and (iv) ultra short down-regulated. Genes coding for proteins with functional relation were mostly cumulated in the same cluster, pointing to a characteristic expression profile for distinct cellular functions. Among the strongest up-regulated genes lpiA, degP1, cah, exoV and exoH were found. The most striking functional groups responding to the shift to acidic pH were genes of the exopolysaccharide I biosynthesis as well as flagellar and chemotaxis genes. While the genes of the exopolysaccharide I biosynthesis (exoY, exoQ, exoW, exoV, exoT, exoH, exoK exoL, exoO, exoN, exoP) were up-regulated, the expression level of the flagellar and chemotaxis genes (visR, motA, flgF, flgB, flgC, fliE, flgG, flgE, flgL, flbT, mcpU) simultaneously decreased in response to acidic pH. Other responding functional groups of genes mainly belonged to nitrogen uptake and metabolism (amtB, nrtB, nirB, nirD), methionine metabolism (metA, metF, metH, metK, bmt and ahcY) as well as ion transport systems (sitABCD, phoCD). It is noteworthy, that several genes coding for hypothetical proteins of unknown function could be identified as up-regulated in response to the pH shift. CONCLUSION: It was shown that the short term response to acidic pH stress does not result in a simple induction or repression of genes, but in a sequence of responses varying in their intensity over time. Obviously, the response to acidic pH is not based on a few specific genes, but involves whole sets of genes associated with various cellular functions. PMID- 19216802 TI - Human papillomavirus type 16 variants in cervical intraepithelial neoplasia and invasive carcinoma in San Luis Potosi City, Mexico. AB - BACKGROUND: In San Luis Potosi City cervical infection by human papillomavirus type 16 (HPV16) associated to dysplastic lesions is more prevalent in younger women. In this work HPV16 subtypes and variants associated to low-grade intraepithelial lesions (LSIL), high-grade intraepithelial lesions (HSIL) and invasive cervical cancer (ICC) of 38 women residing in San Luis Potosi City were identified by comparing their E6 open reading frame sequences. RESULTS: Three European (E) variants (E-P, n = 27; E-T350G, n = 7; E-C188G, n = 2) and one AA-a variant (n = 2) were identified among the 38 HPV16 sequences analyzed. E-P variant sequences contained 23 single nucleotide changes, two of which (A334G, A404T) had not been described before and allowed the phylogenetic separation from the other variants. E-P A334G sequences were the most prevalent (22 cases, 57.9%), followed by the E-P Ref prototype (8 cases, 21.1%) and E-P A404T (1 case, 2.6%) sequences. The HSIL + ICC fraction was 0.21 for the E-P A334G variants and 0.00 for the E-P Ref variants. CONCLUSION: We conclude that in the women included in this study the HPV16 E subtype is 19 times more frequent than the AA subtype; that the circulating E variants are E-P (71.1%) > E-T350G (18.4%) > E-C188G (5.3%); that 71.0% of the E-P sequences carry the A334G single nucleotide change and appear to correspond to a HPV16 variant characteristic of San Luis Potosi City more oncogenic than the E-P Ref prototype. PMID- 19216803 TI - Expression of nicotinamide N-methyltransferase in hepatocellular carcinoma is associated with poor prognosis. AB - BACKGROUND: Hepatocellular carcinoma (HCC) is the most common tumor in the adult liver, with high relapse and mortality rates despite diverse treatment modalities. In this study, nicotinamide N-methyltransferase (NNMT), a key enzyme in drug metabolism, was investigated as a potential prognostic factor. METHODS: Frozen tumors and non-cancerous surrounding tissues from 120 patients with primary HCC were studied. Expressions of NNMT and internal control genes were measured by real-time reverse-transcription PCR (RT-PCR). The relationship of NNMT mRNA level with clinicopathologic parameters and clinical outcome was evaluated. RESULTS: NNMT mRNA level is markedly reduced in HCCs compared to non cancerous surrounding tissues (P < 0.0001), and NNMT expression in tumors was significantly correlated with tumor stage (P = 0.010). Moreover, stratification of patients based on tumor NNMT mRNA levels revealed that the patients who expressed higher NNMT mRNA levels tended to have a shorter overall survival (OS) time (P = 0.053) and a significantly shorter disease-free survival (DFS) time (P = 0.016). Both NNMT expression (P = 0.0096) and tumor stage (P = 0.0017) were found to be significant prognostic factors for DFS in a multivariate analysis. CONCLUSION: The results of this study indicated that NNMT gene expression is associated with tumor stage and DFS time in HCC cases. Because of the broad substrate specificity of NNMT, which could alter the efficacy and adverse effects of chemotherapy, NNMT merits further investigation regarding its role as a prognostic factor with a larger cohort of HCC patients. PMID- 19216804 TI - Dynamic distribution and expression in vivo of the human interferon gamma gene delivered by adenoviral vector. AB - BACKGROUND: We previously found that r-hu-IFNgamma exerts a potent anti-tumor effect on human nasopharyngeal carcinoma xenografts in vivo. Considering the fact that the clinical use of recombinant IFNgamma is limited by its short half-life and systemic side effects, we developed a recombinant adenovirus, Ad-IFNgamma. METHODS: Dynamic distribution of the adenovirus vector and expression of IFNgamma were evaluated by Q-PCR and ELISA after intratumoral administration of Ad IFNgamma into CNE-2 xenografts. RESULTS: Ad-IFNgamma DNA was mainly enriched in tumors where the Ad-IFNgamma DNA was injected (P < 0.05, compared to blood or parenchymal organs), as well as in livers (P < 0.05). Concentrations of Ad IFNgamma DNA in other organs and blood were very low. Intratumoral Ad-IFNgamma DNA decreased sharply at high concentrations (9 x 10(5) copies/microg tissue DNA), and slowly at lower concentrations (1.7-2.9 x 10(5) copies/microg tissue DNA). IFNgamma was detected in the tumors and parenchymal organs. The concentration of IFNgamma was highest in the tumor (P < 0.05), followed by the liver and kidney (P < 0.05). High-level intratumoral expression of IFNgamma was maintained for at least 7 days, rapidly peaking on day 3 after injection of Ad IFNgamma DNA. CONCLUSION: An IFNgamma gene delivered by an adenoviral vector achieved high and consistent intratumoral expression. Disseminated Ad-IFNgamma DNA and the transgene product were mainly enriched in the liver. PMID- 19216805 TI - Role of oxidative stress and intracellular glutathione in the sensitivity to apoptosis induced by proteasome inhibitor in thyroid cancer cells. AB - BACKGROUND: The proteasome inhibitor bortezomib has shown impressive clinical activity alone and in combination with conventional and other novel agents for the treatment of multiple myeloma (MM) and some solid cancers. Although bortezomib is known to be a selective proteasome inhibitor, the downstream mechanisms of cytotoxicity and drug resistance are poorly understood. METHODS: Proteasome activity, intracellular glutathione (GSH) and ROS levels, as well as activities of GSH synthesis enzymes were measured using spectrophotometric methods. Cell death was analyzed using flow cytometry and caspase activity assay. The expression level of GSH synthesis enzymes were measured using real-time RT PCR. RESULTS: At concentrations that effectively inhibited proteasome activity, bortezomib induced apoptosis in FRO cells, but not in ARO cells. Bortezomib elevated the amount of glutathione (GSH) and the treatment with bortezomib increased the level of mRNA for GCL, a rate-limiting enzyme in glutathione synthesis. Furthermore, depletion of GSH increases apoptosis induced by bortezomib, in contrast, repletion of GSH decreases bortezomib-mediated cell death. CONCLUSION: GSH protects cells from proteasome inhibition-induced oxidative stress and glutathione-dependent redox system might play an important role in the sensitivity to proteasome inhibition-induced apoptosis. PMID- 19216806 TI - Detection of lymphangiogenesis in non-small cell lung cancer and its prognostic value. AB - BACKGROUND: Our aim was to detect lymphatic endothelial marker podoplanin, lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1) and vascular endothelial growth factor receptor-3 (VEGFR)-3 and study the prognostic relevance of lymphangiogenesis in non-small cell lung cancer (NSCLC). MATERIALS: 82 paraffin-embedded tissues and 40 fresh frozen tissues from patients with NSCLC were studied. Tumor samples were immunostained for the lymphatic endothelial markers. Lymphangiogenesis was assessed by immunohistochemical double stains for Podoplanin and Ki-67. The prognostic relevance of lymphangiogenesis-related clinicopathological parameters in NSCLC was evaluated. RESULTS: We found that the number of podoplanin positive vessels was correlated positively with the number of LYVE-1 positive vessels. Most of VEGFR-3 positive, few of LYVE-1 positive and none of podoplanin positive vessels were blood vessels. Peritumoral lymphatic vessel density (ptLVD), pathologic stage, lymph node status, lymphatic vessel invasion (LVI), vascular endothelial growth factor-C (VEGF-C) expression and Ki 67 index of the endothelium cells of the micro lymphatic vessels (Ki67%) were associated significantly with a higher risk of tumor progress. ptLVD, pathologic stage, lymph-node metastasis and Ki67% were independent prognostic parameters for overall survival. CONCLUSION: Podoplanin positive ptLVD might play important roles in the lymphangiogenesis and progression of NSCLC. Patients with high podoplanin+ ptLVD have a poor prognosis. PMID- 19216807 TI - Children's and parents' perceptions of the determinants of children's fruit and vegetable intake in a low-intake population. AB - OBJECTIVE: To study the differences between children's self-reports and parents' reports on environmental determinants of fruit and vegetable intake among 11-year old children in the European country with the lowest reported consumption. A second objective was to examine the observed variance in fruit and vegetable intake among the children. DESIGN: A cross-sectional survey was performed in Iceland as a part of the Pro Children cross-Europe survey. Children's usual fruit and vegetable intake and its determinants were assessed through self-reports from the children (n 963) and their parents' reports. RESULTS: Children reported lower availability and accessibility of fruits at home than did their parents, while the reports of children and parents for vegetables were more in agreement. A larger proportion of the observed variance in children's fruit and vegetable intake could be explained by the child's perception than by the parent's perception of determinants. The strongest determinants for fruit and vegetable intake according to the children's reports were availability at home, modelling, demanding family rule and knowledge of recommendations. The strongest modelling determinant for fruit was the father's fruit intake while for vegetables it was eating vegetables together with the family. CONCLUSION: Eleven-year-old children should be asked themselves what determines their fruit and vegetable intake. However, children reported determinants in the physical and social environment, of which the parents are a part, as an important determinant for their intake. Interventions aiming to increase fruit and vegetable intake among children must therefore target the parents. PMID- 19216808 TI - Lutein concentration in human milk during early lactation and its relationship with dietary lutein intake. AB - OBJECTIVE: The present study aimed to estimate the lutein concentration in human milk during early lactation and its relationship with dietary lutein intake measured through the administration of a short FFQ. DESIGN: A cross-sectional study in which an FFQ was administered twice: on day 3 (T0) and day 30 (T1) postpartum; meanwhile two breast milk samples were collected. Maternal plasma samples were obtained at T0. The comparison of dietary lutein intakes and likewise lutein concentrations in breast milk at T0 and T1 were analysed with Student's t test. Pearson's correlation coefficient was used to determine the association between dietary lutein intake and lutein concentration in milk and plasma, respectively, as well as the correlation between breast milk and plasma lutein concentrations at T0. SETTING: Pavia, northern Italy. SUBJECTS: Twenty-one pregnant women, age range 24-42 years, were recruited during their last trimester on a voluntary basis. RESULTS: Both breast milk and plasma lutein concentrations were significantly correlated with dietary lutein intake (r = 0.86, P = 0.0001 and r = 0.94, P = 0.0001, respectively). There was a clear significant correlation between milk and plasma lutein concentrations (r = 0.87, P = 0.0001). Mature milk lutein concentration, although significantly reduced at T1 (P < 0.01), maintained a fairly high correlation with dietary lutein intake (r = 0.82, P = 0.0001). CONCLUSIONS: Even though milk lutein concentration decreased during early lactation, it remained significantly correlated with daily lutein intake. Therefore, while awaiting further research, dietary recommendations advising intake of fresh fruit and vegetables rich in lutein, throughout the whole duration of pregnancy and lactation, are extremely useful. PMID- 19216810 TI - Development of a performance-rating scale for a nutrition knowledge test developed for adolescents. AB - OBJECTIVE: The objectives of the present study were to (i) develop and validate a norm-referenced performance-rating scale to interpret a nutrition knowledge test developed for urban adolescents and (ii) develop a prototype for other researchers to follow when developing nutrition knowledge tests. DESIGN: For norm development the nutrition knowledge test (questionnaire) was administered to a sample representative of the questionnaire target group, referred to as the norm group. These included 512 adolescents in grades 8 (n 158), 10 (n 149) and 12 (n 205) at three randomly selected schools in Soweto and Johannesburg. The performance scores (in percentages) obtained by the norm group were transformed to Z-scores which were categorised into stanines using established Z-score cut off points. For validation purposes the questionnaire was completed by 148 volunteers: sixty university dietetics students, nineteen non-nutrition university students and sixty-nine primary-school teachers. RESULTS: As required of an ideal norm group, the Z-scores formed a normal distribution (a bell-shaped curve). To facilitate interpretation of the results, the Z-score cut-off points for these categories were transformed back to performance scores (percentages) so that the performance of a testee could be interpreted directly from his/her performance in percentage. As is recommended, the nine stanine categories were reduced to five: very poor, fair/below average, good/average, very good/above average and excellent. The discriminatory validity of the norms was substantiated by showing that groups with known nutrition knowledge levels were rated appropriately and that the performance ratings of these groups differed significantly, with university dietetics students scoring 98.3%, primary-school teachers 20.3% and non-nutrition university students 31.6%. CONCLUSIONS: The norm referenced performance-rating scale can be used with confidence to interpret the performance score achieved by a testee on the nutrition knowledge test developed for urban adolescents in South Africa. The methodology used in the study serves as a prototype for other researchers who are developing knowledge tests. PMID- 19216809 TI - Habitual high phosphorus intakes and foods with phosphate additives negatively affect serum parathyroid hormone concentration: a cross-sectional study on healthy premenopausal women. AB - OBJECTIVE: Foods can contain natural phosphorus (NP) and phosphate-containing food additives (AP). The main objective of the present study was to investigate whether NP and AP of habitual diets differ in their effects on markers of Ca metabolism. We also investigated the impact of total habitual dietary P intake on markers of Ca metabolism. DESIGN: Cross-sectional study. Fasting blood samples were collected and participants kept a 4 d food record, from which dietary intake of total P and the consumption of NP (milk and cheese, excluding processed cheese) and AP (processed cheese) sources were calculated. Participants were divided into groups according to their NP- and AP-containing food consumption and into quartiles according to their total P intake. SETTING: Southern Finland. SUBJECTS: One hundred and forty-seven healthy premenopausal women aged 31-43 years. RESULTS: Relative to the lowest total dietary P quartile, mean serum parathyroid hormone (S-PTH) concentration was higher (P = 0.048, analysis of covariance (ANCOVA)) and the mean serum ionized Ca concentration lower (P = 0.016, ANCOVA) in the highest P intake quartile. Mean S-PTH concentrations were higher among participants who consumed processed cheese (P = 0.027, ANCOVA) and less milk and other cheese than processed cheese (P = 0.030, ANCOVA). CONCLUSIONS: High total habitual dietary P intake affected S-PTH unfavourably. Furthermore, phosphate additives may have more harmful effects on bone than other P sources, as indicated by higher mean S-PTH concentration among participants who consumed AP-containing foods. Because of the high dietary P intake and current upward trend in consumption of processed foods in Western countries, these findings may have important public health implications. PMID- 19216811 TI - A social marketing approach to improving the nutrition of low-income women and children: an initial focus group study. AB - OBJECTIVE: To identify approaches for interventions to improve the nutrition of low-income women and children. DESIGN: Seven focus groups were conducted with low income women caring for young children in their households. They discussed shopping, eating at home, eating out and healthy eating. The discussions were recorded and subjected to qualitative thematic analysis. SETTING: A semi-rural community in Oregon, USA. SUBJECTS: There were seventy-four women (74% White), most of whom were 18-29 years old. RESULTS: Four broad themes were identified, i.e. cost-consciousness, convenience, social influences and health issues. CONCLUSIONS: The target population would benefit from improved understanding of what constitutes a balanced diet, with a greater emphasis on a more central role for fruit and vegetables. To persuade this population to change their eating habits, it will be necessary to convince them that healthful food can be low cost, convenient and palatable for children. Comparing findings from the present study with a similar one in the UK suggests that the US women faced many of the same barriers to healthy eating but displayed less helplessness. PMID- 19216812 TI - Effects of the flaxseed lignans secoisolariciresinol diglucoside and its aglycone on serum and hepatic lipids in hyperlipidaemic rats. AB - The present study involved a comparative analysis of the effects of purified flaxseed lignans, secoisolariciresinol diglucoside (SDG) and its aglycone metabolite (SECO), in hyperlipidaemic rats. For hypercholesterolaemia, female Wistars (six rats per group) were fed a standard or 1 % cholesterol diet and orally administered 0, 3 or 6 mg SDG/kg or 0, 1.6 or 3.2 mg SECO/kg body weight once daily for 4 weeks. Hypertriacylglycerolaemia was induced in male Sprague Dawley rats (ten rats per group) by supplementing tap water with 10 % fructose. These rats were orally administered 0, 3 or 6 mg SDG/kg body weight once daily for 2 weeks. Fasting blood samples (12 h) were collected predose and at the end of the dosing period for serum lipid analyses. Rats were killed and livers rapidly excised and sectioned for lipid, mRNA and histological analyses. Chronic administration of equimolar amounts of SDG and SECO caused similar dose-dependent reductions in rate of body-weight gain and in serum total and LDL-cholesterol levels and hepatic lipid accumulation. SDG and SECO failed to alter hepatic gene expression of commonly reported regulatory targets of lipid homeostasis. SDG had no effect on serum TAG, NEFA, phospholipids and rate of weight gain in 10 % fructose-supplemented rats. In conclusion, our data suggest that the lignan component of flaxseed contributes to the hypocholesterolaemic effects of flaxseed consumption observed in humans. Future studies plan to identify the biochemical mechanism(s) through which flaxseed lignans exert their beneficial effects and the lignan form(s) responsible. PMID- 19216813 TI - Increased portion size leads to a sustained increase in energy intake over 4 d in normal-weight and overweight men and women. AB - Large food portions may be facilitating excess energy intake (EI) and adiposity among adults. The present study aimed to assess the extent to which EI and amounts of foods consumed are influenced by the availability of different-sized food portions. A randomised within-subject cross-over, fully residential design was used, where forty-three (twenty-one men and twenty-two women) normal-weight and overweight adults were randomly allocated to two separate 4 d periods where they were presented with either 'standard' or 'large' food portions of the same foods and beverages. The main outcome measures were the amount of food (g) and EI (MJ) consumed throughout each study period. Mean EI over 4 d was significantly higher on the large portion condition compared with the standard condition in the total group (59.1 (sd 6.6) v. 52.2 (sd 14.3) MJ; P = 0.020); men and women increased their EI by 17 % (10 (sd 6.5) MJ; P < 0.001) and 10 % (4 (sd 6.5) MJ; P = 0.005) respectively when served the large food portions relative to the standard food portions. The increased intakes were sustained over the 4 d in the large portion condition with little evidence of down-regulation of EI and food intake being made by subjects. Increased food portion size resulted in significant and sustained increases in EI in men and women over 4 d under fully residential conditions. The availability and consumption of larger portions of food may be a significant factor contributing to excess EI and adiposity. PMID- 19216814 TI - A high-protein, moderate-energy, regular cheesy snack is energetically compensated in human subjects. AB - Snacking is often regarded as a cause of overweight. However, the main issue is to determine whether the consumption of snacks leads to an increase in energy intake or whether a compensation phenomenon exists and maintains daily energy intake at a constant level. The objective of the present study was to determine whether the repeated consumption of a high-protein, moderate-energy, cheesy snack given as a preload 1 h before a meal altered energy intake at the next meal and then throughout the day, and if this kind of snack was energetically compensated. Normal-weight women (n 27) were recruited for the study. All subjects were healthy non-smokers, aged 18-60 years. The snacks consisted of portions of cheese containing 22 g protein, with an energy value of 836 kJ. Two types of snack were compared, differing in terms of the type of milk proteins they contained: the first contained casein only (CAS), while the second contained a mixture of casein and whey proteins (WHEY+CAS; 2:1). The principal finding of the present study was that the ingestion of the two snacks 1 h before lunch led to energy compensation of 83.1 (SEM 9.4) and 67.0 (SEM 16.4) % for WHEY+CAS and CAS respectively, at lunch, and 121.6 (SEM 36.5) and 142.1 (SEM 29.7) % for WHEY+CAS and CAS respectively, regarding the whole-day energy intake. In conclusion, the repeated consumption of a high-protein, moderate-energy, regular cheesy snack should not promote overweight because energy intake appears to be regulated during subsequent meals on the same day. PMID- 19216815 TI - Computed tomography and/or magnetic resonance imaging for pre-operative planning for inverted nasal papilloma: review of evidence. AB - BACKGROUND: Inverted nasal papilloma is a benign tumour with variable biological behaviour. It is a unique neoplasm and is often difficult to manage, being characterised by a tendency to recur following excision, an association with malignancy and an ability to destroy bone. Radiological diagnosis has traditionally been based on computed tomography, but it is often impossible to differentiate between polyps with entrapped debris and inverted nasal papilloma. Magnetic resonance imaging, especially T2-weighted images, is perhaps a better tool in differentiating inverted nasal papilloma from other nasal lesions, and has recently been advocated as the imaging modality of choice. AIMS: To review the evidence on the ideal imaging modality for pre-operative planning of surgery for patients with histologically proven inverted nasal papilloma. MATERIALS AND METHODS: A systematic review of studies was undertaken, focusing on radiological assessment of inverted nasal papilloma (primary and recurrent). All English language articles were potentially included in the review. However, we excluded single case reports, case series, pictorial essays, 'teaching' reviews and reviews of inverted nasal papilloma not focusing on pre-operative imaging. RESULTS: Sixteen studies were identified based on our search strategy. Only 10 fulfilled our criteria. Computed tomography scanning was the standard imaging modality of choice for assessing sinus involvement of inverted papilloma and for planning the extent of surgery. T2-weighted magnetic resonance imaging scans were able to distinguish tumour (intermediate signal) from inflammatory tissues (bright signal), but not post-operative scarring from recurrent tumour. No studies were found which compared the specificity and sensitivity of magnetic resonance imaging and computed tomography in the accurate pre-operative identification of the extent of inverted papilloma. CONCLUSIONS: There is currently not enough evidence to suggest one sole modality as providing optimum imaging for inverted nasal papilloma. Computed tomography remains the imaging modality of first choice for inverted nasal papilloma, despite certain disadvantages. Magnetic resonance imaging is able to distinguish tumour from inflammation and is advocated as a better tool for recurrent tumour, but bone destruction of sinus walls is less easy to recognise, compared with computed tomography. Evaluation of sinus tumours usually involves both imaging modalities, and inverted nasal papilloma should be included within this pathological group. The cohort of patients is usually small, so cost-effectiveness should not generally be an issue when considering whether to use computed tomography, magnetic resonance imaging or both. A well structured, prospective study is needed to evaluate the efficacy of magnetic resonance imaging versus computed tomography for pre-operative planning of histologically proven inverted nasal papilloma. PMID- 19216816 TI - Anatomical variation of internal carotid artery presenting as pharyngeal mass. AB - OBJECTIVE: We describe a case of an internal carotid artery loop presenting as an oropharyngeal mass. METHOD: Case report and review of current literature. CASE REPORT: A 71-year-old woman presented with an asymptomatic oropharyngeal mass. This was an incidental finding by her general practitioner and was urgently referred as a suspicious lesion. A magnetic resonance imaging scan revealed an internal carotid artery tonsillar kink indenting into the pharyngeal wall. No further treatment was necessary. CONCLUSION: The internal carotid kink is an important anatomical variation which ENT surgeons should be aware of, as there are significant complications if blind biopsy is undertaken prior to appropriate imaging. PMID- 19216817 TI - Peroral drainage of post-traumatic sialocoeles: report of three cases. AB - OBJECTIVE: To discuss the technique and outcome of this simple procedure and the management of post-traumatic parotid sialocoeles, and to review the literature regarding this condition. CASE REPORT: We report the successful surgical treatment, by peroral drainage, of three patients with post-traumatic parotid sialocoele resistant to conservative management. DISCUSSION: We discuss the method and outcome of the surgical procedure performed, along with the causes, presentation and management of parotid sialocoele. CONCLUSION: Correct initial management of a parotid duct injury may prevent the formation of a sialocoele. When conservative treatment of post-traumatic parotid sialocoele fails, we advocate the surgical technique described in this report as it is effective, simple and carries minimal risk to the patient. PMID- 19216818 TI - Topical chemoprevention of skin cancer in mice, using combined inhibitors of 5 lipoxygenase and cyclo-oxygenase-2. AB - OBJECTIVE: Skin cancer is the most common cancer, and often occurs in the head and neck region. This study aimed to investigate whether a combination of inhibitors of cyclo-oxygenase-2 and 5-lipoxygenase, applied via a microemulsion delivery system, would be effective in topically inhibiting skin carcinogenesis. STUDY DESIGN: Randomised animal study. METHODS: Twenty-four nude mice were intradermally inoculated with carcinoma cells and then divided into three groups (eight animals each): group one received no treatment; group two received celecoxib alone; and group three received a combination of zileuton and celecoxib. Tumour incidence and growth were measured for 14 days. RESULTS: Both treatments significantly delayed the onset and development of tumours. However, the combined treatment had the best response (p < 0.01). CONCLUSION: The results clearly showed that topical treatment with either celecoxib alone or celecoxib plus zileuton significantly inhibited skin carcinogenesis, and that a combination of both agents had the best results. PMID- 19216819 TI - Contour statistics in natural images: grouping across occlusions. AB - Correctly interpreting a natural image requires dealing properly with the effects of occlusion, and hence, contour grouping across occlusions is a major component of many natural visual tasks. To better understand the mechanisms of contour grouping across occlusions, we (a) measured the pair-wise statistics of edge elements from contours in natural images, as a function of edge element geometry and contrast polarity, (b) derived the ideal Bayesian observer for a contour occlusion task where the stimuli were extracted directly from natural images, and then (c) measured human performance in the same contour occlusion task. In addition to discovering new statistical properties of natural contours, we found that naive human observers closely parallel ideal performance in our contour occlusion task. In fact, there was no region of the four-dimensional stimulus space (three geometry dimensions and one contrast dimension) where humans did not closely parallel the performance of the ideal observer (i.e., efficiency was approximately constant over the entire space). These results reject many other contour grouping hypotheses and strongly suggest that the neural mechanisms of contour grouping are tightly related to the statistical properties of contours in natural images. PMID- 19216820 TI - How many pixels make an image? AB - The human visual system is remarkably tolerant to degradation in image resolution: human performance in scene categorization remains high no matter whether low-resolution images or multimegapixel images are used. This observation raises the question of how many pixels are required to form a meaningful representation of an image and identify the objects it contains. In this article, we show that very small thumbnail images at the spatial resolution of 32 x 32 color pixels provide enough information to identify the semantic category of real world scenes. Most strikingly, this low resolution permits observers to report, with 80% accuracy, four to five of the objects that the scene contains, despite the fact that some of these objects are unrecognizable in isolation. The robustness of the information available at very low resolution for describing semantic content of natural images could be an important asset to explain the speed and efficiently at which the human brain comprehends the gist of visual scenes. PMID- 19216821 TI - Validation of the 10/66 Dementia Research Group's 10/66 Dementia diagnosis in Iran. PMID- 19216822 TI - Is there a role for psychostimulants in old age depression and apathy? PMID- 19216823 TI - Heterorhabditis gerrardi n. sp. (Nematoda: Heterorhabditidae): the hidden host of Photorhabdus asymbiotica (Enterobacteriaceae: gamma-Proteobacteria). AB - A new entomopathogenic nematode species from Australia, Heterorhabditis gerrardi n. sp. (Nematoda: Heterorhabditidae) is described. Morphological and molecular studies together with cross-hybridization tests indicated that this nematode represents a new undescribed species, closely related to members in the 'indica group'. However, the new species can be distinguished from other species in this genus by a combination of several qualitative and quantitative morphological traits. Key diagnostic features include: body size and excretory pore position of the third-stage infective juveniles; male bursa with a reduction of bursal rays, usually affecting the terminal set of papillae, with symmetrical or asymmetrical loss of one or two pairs; vulva of hermaphrodites more anteriorly located than in other species in the indica-group (V% average: 43), with non-protruding or slightly protruding lips, and longer tail length (average: 106 mum). The new species can be further characterized by molecular traits of sequence data from the internal transcribed spacer (ITS) region of ribosomal DNA. Additionally, the bacterial symbiont of this new species, Photorhabdus asymbiotica Kingscliff strain, was phenotypically characterized and compared with other P. asymbiotica strains. The Kingscliff strain revealed many characters not present in other strains of this species. We hypothesize that the newly found traits may contribute to the maintenance of this mutualistic association of the bacterium with its nematode host. PMID- 19216824 TI - Causes of intraspecific variation in body size among trematode metacercariae. AB - Inequalities in body size among adult helminths can result in inequalities in reproductive output, with consequences for population dynamics and genetics. These inequalities can result from growth differences among larval worms inside intermediate hosts that persist into the adult stage. Here, we investigate the effects of both host body size and intensity of infection on the sizes of metacercariae of the trematode Maritrema novaezealandensis (Microphallidae) inside their second intermediate host, the isopod Paridotea ungulata (Idoteidae). Among the more than 1500 metacercariae recovered and individually measured, there was no relationship between the mean diameter of metacercarial cysts per isopod and isopod body length. However, intensity of infection correlated negatively with the mean diameter of cysts within an isopod, i.e. metacercariae in crowded infections attained smaller sizes on average. In contrast, the variability in cyst sizes per isopod, measured as the coefficient of variation, was independent of both isopod body length and infection intensity. Our results show that a disproportionate number of relatively small metacercariae come from the relatively few hosts in which a large fraction of all metacercariae are aggregated. The combination of aggregation and intensity-dependent growth generates inequalities in sizes among metacercariae that will be passed on to adult worm populations in definitive hosts. PMID- 19216825 TI - Predatory activity of the fungi Duddingtonia flagrans, Monacrosporium thaumasium, Monacrosporium sinense and Arthrobotrys robusta on Angiostrongylus vasorum first stage larvae. AB - Angiostrongylus vasorum is a nematode that parasitizes domestic dogs and wild canids. We compared the predatory capacity of isolates from the predatory fungi Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34), Monacrosporium sinense (SF53) and Arthrobotrys robusta (I31) on first-stage larvae (L1) of A. vasorum under laboratory conditions. L1 A. vasorum were plated on 2% water-agar (WA) Petri dishes marked into 4 mm diameter fields with the four grown isolates and a control without fungus. Plates of treated groups contained each 1000 L1 A. vasorum and 1000 conidia of the fungal isolates AC001, NF34, SF53 and I31 on 2% WA. Plates of the control group (without fungus) contained only 1000 L1 A. vasorum on 2% WA. Ten random fields (4 mm diameter) were examined per plate of treated and control groups, every 24 h for 7 days. Nematophagous fungi were not observed in the control group during the experiment. There was no variation in the predatory capacity among the tested fungal isolates (P>0.05) during the 7 days of the experiment. There was a significant reduction (P < 0.05) of 80.3%, 74.5%, 74.2% and 71.8% in the means of A. vasorum L1 recovered from treatments with isolates AC001, NF34, SF53 and I31, respectively, compared to the control without fungi. In this study, the four isolates of predatory fungi were efficient in the in vitro capture and destruction of A. vasorum L1, confirming previous work on the efficiency of nematophagous fungi in the control of nematode parasites of dogs and as a possible alternative method of biological control. PMID- 19216826 TI - Tick-induced blood loss leads to regenerative anaemia in the European hedgehog ( Erinaceus europaeus). AB - Although there is an increasing understanding of the role of parasites in their host dynamics, accurate, quantitative estimates of parasite caused morbidity in wild animals are rare. Here, we examine the possible impact of 2 tick species (Ixodes ricinus, I. hexagonus) on the condition of the European hedgehog (Erinaceus europaeus). For this, we tested for correlations between blood parameters of 36 adult hedgehogs from an experimental population enclosed in a natural habitat and their tick infestation over a period of 8 months (March October 2007). We found correlations between the tick infestation and the concentration of red blood cells, haemoglobin, haematocrit, MCH, MCHC, thrombocytes, lymphocytes and neutrophils. These results indicate that ticks can induce anaemia in the hedgehog. The peripheral blood characteristics and the erythrocyte indices characterize this anaemia as haemorrhagic and regenerative. During the course of our study the hedgehogs of our population showed below normal mortality but morbidity was found to be high resulting from the blood loss caused by the feeding activity of the ticks. PMID- 19216827 TI - Trypanosoma caninum n. sp. (Protozoa: Kinetoplastida) isolated from intact skin of a domestic dog ( Canis familiaris) captured in Rio de Janeiro, Brazil. AB - An unknown Trypanosoma species was isolated from an axenic culture of intact skin from a domestic dog captured in Rio de Janeiro, Brazil, which was co-infected with Leishmania (Viannia) braziliensis. Giemsa-stained smears of cultures grown in different media revealed the presence of epimastigotes, trypomastigotes, spheromastigotes, transitional stages, and dividing forms (epimastigotes or spheromastigotes). The highest frequency of trypomastigotes was observed in RPMI (15.2%) and DMEM (9.2%) media containing 5% FCS, with a mean length of these forms of 43.0 and 36.0 mum, respectively. Molecular analysis by sequential application of PCR assays indicated that this trypanosome differs from Trypanosoma cruzi and T. rangeli when specific primers were applied. On the other hand, a PCR strategy targeted to the D7 domain of 24salpha rDNA, using primers D75/D76, amplified products of about 250 bp in that isolate (stock A-27), different from the amplification products obtained with T. cruzi and T. rangeli. This organism differs from T. cruzi mainly by the size of its trypomastigote forms and kinetoplasts and the absence of infectivity for macrophages and triatomine bugs. It is also morphologically distinct from salivarian trypanosomes reported in Brazil. Isoenzyme analysis at 8 loci demonstrated a very peculiar banding pattern clearly distinct from those of T. rangeli and T. cruzi. We conclude that this isolate is a new Trypanosoma species. The name T. caninum is suggested. PMID- 19216829 TI - Vaccenic acid favourably alters immune function in obese JCR:LA-cp rats. AB - Vaccenic acid (VA) is a ruminant-derived trans-fat and precursor of conjugated linoleic acid (CLA). The objective of the present study was to explore the effects of VA on immune function in a model of the metabolic syndrome, JCR:LA-cp rats. Lean (2:1 mix of +/cp and +/+) and obese (cp/cp) rats, aged 8 weeks, were fed a control (0% VA) or a VA diet (1.5% (w/w) VA) for 3 weeks (twenty rats per group). Splenocytes and mesenteric lymph node (MLN) immune cell phenotypes (flow cytometry), ex vivo cytokine production (ELISA) and phospholipid fatty acid concentrations were measured. Obese rats had higher proportions of splenic macrophages, total T-cells, helper T-cells (total and percentage CD25+), cytotoxic T-cells (total and percentage CD25+) and produced higher concentrations of IL-6 to concanavalin A (ConA) compared with lean rats. Obese rats had lower proportions of MLN T-cells, new T-cells (CD3+CD90+) and cytotoxic T-cells, but higher proportions of helper cells that were CD45RC+, CD25+ and CD4lo, and produced higher concentrations of IL-2, IL-10, interferon gamma and TNFalpha in response to ConA compared with lean rats. VA was higher in plasma phospholipids and both VA and CLA (cis-9, trans-11) were higher in MLN phospholipids compared with control-fed rats. Lean VA-fed rats had lower proportions of MLN and splenocyte CD45RC+ helper cells, and helper T-cells. Splenocytes from VA-fed rats produced 16-23% less IL-2, IL-10 and TNFalpha compared with controls. VA normalised production of MLN IL-2 and TNFalpha in obese rats to levels similar to those seen in lean rats. These results indicate that dietary VA favourably alters the pro-inflammatory tendency of mesenteric lymphocytes from JCR:LA-cp rats. PMID- 19216828 TI - Cross-sectional association of dietary patterns with insulin-resistant phenotypes among adults without diabetes in the Framingham Offspring Study. AB - Cluster analysis is a valuable tool for exploring the health consequences of consuming different dietary patterns. We used this approach to examine the cross sectional relationship between dietary patterns and insulin-resistant phenotypes, including waist circumference, BMI, fasting insulin, 2 h post-challenge insulin, insulin sensitivity index (ISI0,120), HDL-cholesterol, TAG and blood pressure, using data from the fifth examination cycle of the Framingham Offspring Study. Among 2875 participants without diabetes, we identified four dietary patterns based on the predominant sources of energy: 'Fruits, Reduced Fat Dairy and Whole Grains', 'Refined Grains and Sweets', 'Beer' and 'Soda'. After adjusting for multiple comparisons and potential confounders, compared with the 'Fruits, Reduced Fat Dairy and Whole Grains' pattern, the 'Refined Grains and Sweets' pattern had significantly higher mean waist circumference (92.4 v. 90.5 cm; P = 0.008) and BMI (27.3 v. 26.6 kg/m2; P = 0.02); the 'Soda' pattern had significantly higher mean fasting insulin concentration (31.3 v. 28.0 microU/ml; P < or = 0.001); the 'Beer' pattern had significantly higher mean HDL-cholesterol concentration (1.46 v. 1.31 mmol/l; P < 0.001). No associations were observed between dietary patterns and ISI0,120, TAG, and systolic or diastolic blood pressure. Our findings suggest that consumption of a diet rich in fruits, vegetables, whole grains and reduced-fat dairy protects against insulin-resistant phenotypes and displacing these healthy choices with refined grains, high-fat dairy, sweet baked foods, candy and sugar-sweetened soda may promote insulin resistant phenotypes. PMID- 19216830 TI - A multi-gene analysis strategy identifies metabolic pathways targeted by trans 10, cis-12-conjugated linoleic acid in the liver of hamsters. AB - In mice, hepatic functions can be greatly affected by dietary trans-10, cis-12 conjugated linoleic acid (CLA). However, this phenomenon has been less documented in hamsters. In the present study, male hamsters were fed two doses of the trans 10, cis-12-CLA (0.5 and 1%, w/w diet) or linoleic acid (0.5%) for 6 weeks. The effects on the liver were examined by measuring the expression of thirty-six genes representing key metabolic pathways. CLA-responsive genes and their relationships with physiological outcomes were examined by a multivariate analysis procedure. Compared with control hamsters, those receiving either 0.5 or 1% CLA exhibited similar fat loss (15-24%; P < or = 0.05) and liver enlargement (21-28%; P < or = 0.05), with no signs of steatosis. We also observed a dose dependent increase in the transcription of genes involved in lipid breakdown and lipid harvesting from blood, and in genes related to the oxidative stress and inflammatory responses. These responsive genes varied in parallel with cell membrane lipids (R2 0.31-0.42) and to a lesser extent with liver enlargement (R2 0.22) (all P < 0.05). We conclude that in hamsters, liver enlargement induced by trans-10, cis-12-CLA is accompanied by an increased metabolic potential to process fatty acids from mobilised adipose stores. This elevated metabolic activity, comprised of anabolic pathways and their catabolic counterparts, can trigger inflammation and the oxidant stress defence pathways in a dose-dependent manner. These results provide novel insights into the mechanisms by which trans 10, cis-12-CLA affects pathways related to liver function. PMID- 19216832 TI - Still waiting for the great leap forward. PMID- 19216833 TI - Is the incorporation of equity considerations into economic evaluation really so simple? A comment on Cookson, Drummond and Weatherly. PMID- 19216834 TI - Explicit incorporation of equity considerations into economic evaluation of public health interventions. AB - Health equity is one of the main avowed objectives of public health policy across the world. Yet economic evaluations in public health (like those in health care more generally) continue to focus on maximizing health gain. Health equity considerations are rarely mentioned. Health economists rely on the quasi egalitarian value judgment that 'a QALY is a QALY'--that is QALYs are equally weighted and the same health outcome is worth the same no matter how it is achieved or to whom it accrues. This value judgment is questionable in many important circumstances in public health. For example, policy-makers may place rather little value on health outcomes achieved by infringing individual liberties or by discriminating on the basis of age, sex, or race. Furthermore, there is evidence that a majority of the general public wish to give greater weight to health gains accruing to children, the severely ill, and, to a lesser extent, the socio-economically disadvantaged. This paper outlines four approaches to explicit incorporation of equity considerations into economic evaluation in public health: (i) review of background information on equity, (ii) health inequality impact assessment, (iii) analysis of the opportunity cost of equity, and (iv) equity weighting of health outcomes. The first three approaches can readily be applied using standard methods of health technology assessment, where suitable data are available; whereas approaches for generating equity weights remain experimental. The potential benefits of considering equity are likely to be largest in cases involving: (a) interventions that target disadvantaged individuals or communities and are also relatively cost-ineffective and (b) interventions to encourage lifestyle change, which may be relatively ineffective among 'hard-to-reach' disadvantaged groups and hence may require re-design to avoid increasing health inequalities. PMID- 19216835 TI - Management of dysphonia in children. AB - BACKGROUND: Dysphonia is common in children, but practice varies considerably regarding what, if any, investigations are performed and how the condition is managed. Although childhood dysphonia is mostly due to non-serious causes such as voice misuse, very serious pathology such as papillomatosis or malignancy needs occasionally to be excluded, and treatable congenital anomalies such as webs and cysts can be missed. Voice clinics and voice therapy services are now well established in most adult health services in the developed world, but equivalent services for children are less common, at least in the UK. METHODS: We retrospectively reviewed the records of all children presenting to our large children's hospital with a primary complaint of dysphonia between January 2001 and October 2007, in order to determine their management, investigations and final diagnosis. RESULTS: We identified 142 children. Case records were found for 137 (97 per cent). Eight-three children were male (61 per cent) and 54 female (39 per cent). Ages ranged from two months to 15 years (median 5.3 years). In 10 children (7 per cent), hoarseness was congenital, presenting as a hoarse, weak cry at birth. In 15 children (11 per cent), onset of hoarseness was related to a specific surgical procedure. The larynx was visualised by mirror alone in 23 children (17 per cent), by awake fibre-optic laryngoscopy in 27 (20 per cent) and by microlaryngoscopy-bronchoscopy under anaesthesia in 42 (31 per cent). Forty children (29 per cent) did not undergo laryngeal visualisation at any time and were diagnosed based on history alone. A further five (4 per cent) were scheduled for direct laryngoscopy but this was not performed due to resolution of symptoms. Voice abuse accounted for 62 (45 per cent) of all diagnoses. CONCLUSIONS: Childhood dysphonia accounts for a large number of referrals. There is considerable variation in how these children are managed. A more structured approach to diagnosis and investigation would be beneficial, perhaps within the setting of a dedicated paediatric voice clinic. PMID- 19216836 TI - Novel method for safe cauterisation of posterior epistaxis. AB - INTRODUCTION: In epistaxis, anterior bleeding points are easily cauterised under direct vision, but those which occur in the posterior nose present a greater challenge. Standard cautery techniques limit simultaneous use of other equipment in the narrow posterior nose. METHODS: This article presents a novel device which combines suction, cautery stick and sheath in one single-handed implement for ease of use. CONCLUSION: This novel, hand-held device for simultaneous suction and safe cautery of posterior epistaxis is both safe and cost-effective. It enables successful treatment by a single operator and is relatively easy to use by the non-skilled, junior trainee. PMID- 19216837 TI - Improving the evaluation of therapeutic interventions in multiple sclerosis: the role of new psychometric methods. AB - OBJECTIVES: In this monograph we examine the added value of new psychometric methods (Rasch measurement and Item Response Theory) over traditional psychometric approaches by comparing and contrasting their psychometric evaluations of existing sets of rating scale data. We have concentrated on Rasch measurement rather than Item Response Theory because we believe that it is the more advantageous method for health measurement from a conceptual, theoretical and practical perspective. Our intention is to provide an authoritative document that describes the principles of Rasch measurement and the practice of Rasch analysis in a clear, detailed, non-technical form that is accurate and accessible to clinicians and researchers in health measurement. REVIEW METHODS: A comparison was undertaken of traditional and new psychometric methods in five large sets of rating scale data: (1) evaluation of the Rivermead Mobility Index (RMI) in data from 666 participants in the Cannabis in Multiple Sclerosis (CAMS) study; (2) evaluation of the Multiple Sclerosis Impact Scale (MSIS-29) in data from 1725 people with multiple sclerosis; (3) evaluation of test-retest reliability of MSIS 29 in data from 150 people with multiple sclerosis; (4) examination of the use of Rasch analysis to equate scales purporting to measure the same health construct in 585 people with multiple sclerosis; and (5) comparison of relative responsiveness of the Barthel Index and Functional Independence Measure in data from 1400 people undergoing neurorehabilitation. RESULTS: Both Rasch measurement and Item Response Theory are conceptually and theoretically superior to traditional psychometric methods. Findings from each of the five studies show that Rasch analysis is empirically superior to traditional psychometric methods for evaluating rating scales, developing rating scales, analysing rating scale data, understanding and measuring stability and change, and understanding the health constructs we seek to quantify. CONCLUSIONS: There is considerable added value in using Rasch analysis rather than traditional psychometric methods in health measurement. Future research directions include the need to reproduce our findings in a range of clinical populations, detailed head-to-head comparisons of Rasch analysis and Item Response Theory, and the application of Rasch analysis to clinical practice. PMID- 19216838 TI - Axitinib treatment in patients with cytokine-refractory metastatic renal cell cancer. PMID- 19216839 TI - Myeloid-derived suppressor cells: a novel therapeutic target. AB - Immunotherapy and angiogenic inhibitors, used alone or in combination with chemotherapy, represent two promising cancer treatment programs. Each is limited by myeloid-derived suppressor cells (MDSCs), which accumulate in tumor-bearing hosts. MDSCs inhibit effector T-cell function and thus prevent the formation and execution of an effective antitumor immune response. Recently reported studies have shown that MDSCs also function to promote tumor-dependent angiogenesis as well as tumor metastasis, and to provide tumor resistance to antiangiogenic drugs. Insights into tumor-imposed dynamic changes in bone marrow function and myeloid cell development should fuel novel drug developments and novel applications of drugs currently in use. Such insights suggest that multitargeted receptor tyrosine kinase inhibitors, such as sunitinib, may be useful adjunctive agents for use in immunotherapy trials treating several tumor types. PMID- 19216840 TI - VHL gene mutations in renal cell carcinoma: role as a biomarker of disease outcome and drug efficacy. AB - The therapeutic landscape for renal cell carcinoma (RCC) has changed drastically over the past several years with the emergence of molecularly targeted therapies. With previous prognostic and predictive tools based on studies of patients treated with cytokine therapies, confirmation of these prior methods and discovery of new markers in this new era of targeted therapy are of great importance. Alteration of the von Hippel-Lindau gene (VHL) by mutation, loss of heterozygosity, and promoter methylation has been found to be important to RCC pathogenesis. In this review, we discuss the potential role of VHL mutation as a prognostic and predictive marker for RCC. PMID- 19216841 TI - Perifosine: update on a novel Akt inhibitor. AB - The PI3K/Akt/mTOR pathway is aberrantly active in most human cancers and contributes to cell growth, proliferation, and survival. Akt is a nodal regulator of cellular survival pathways and an attractive target in cancer therapy. Many inhibitors of Akt are being developed. Perifosine is an oral Akt inhibitor currently being tested in phase 2 clinical trials. Unlike most kinase inhibitors, which target the adenosine triphosphate-binding region, perifosine targets the pleckstrin homology domain of Akt, thereby preventing its translocation to the plasma membrane. Single-agent activity with perifosine has been observed in sarcoma and Waldenstrom macroglobulinemia patients. However, the disappointing response rates of common solid tumors to perifosine as a single agent have diminished expectations and prompted further investigation into its mechanism of action. Perifosine exerts Akt-dependent and Akt-independent effects, and although many preclinical studies have documented Akt inhibition by perifosine, clinical validation of these findings is lacking. In this article, we review the clinical history of perifosine and discuss its many biologic activities. PMID- 19216842 TI - Angiopoietin-2: development of inhibitors for cancer therapy. AB - Angiopoietin-2 (Ang2) is a member of the Ang family, which plays an important role in angiogenesis during the development and growth of human cancers. Ang2's role in angiogenesis generally is considered as an antagonist for Ang1, inhibiting Ang1-promoted Tie2 signaling, which is critical for blood vessel maturation and stabilization. Ang2 modulates angiogenesis in a cooperative manner with another important angiogenic factor, vascular endothelial growth factor A. Genetic studies have revealed that Ang2 also is critical in lymphangiogenesis during development. However, new evidence suggests more complicated roles for Ang2 in angiogenesis in physiologic processes and invasive phenotypes of cancer cells during progression of human cancers. This article discusses recent studies of Ang2 in angiogenesis and the implication of Ang2 as a therapeutic target as well as a potential inhibitor for antiangiogenesis treatment for cancer patients. PMID- 19216843 TI - Jak2 inhibitors: rationale and role as therapeutic agents in hematologic malignancies. AB - Although the Jak2-V617F mutation has generated strong awareness because of its causative role in myeloproliferative disorders, reports of Jak2 gene aberrations linked to hematologic malignancies have preceded those of V617F by nearly a decade. These malignant mutations include Jak2 amino acid substitutions, deletions, insertions, and chromosomal translocations. As a consequence, researchers are increasingly focused on identifying Jak2 inhibitors that suppress aberrant Jak2 kinase activity. Some of these inhibitors may one day become therapeutically beneficial for individuals with Jak2-related hematologic malignancies. This review summarizes various Jak2 mutations associated with hematologic malignancies and assesses some of the Jak2 inhibitors in the preclinical phase or in clinical trials. By reviewing these specific areas, we hope to have a better understanding of Jak2's role in hematologic malignancies and to shed light on the utility of Jak2 inhibitors. PMID- 19216844 TI - The chemokine receptors CXCR4 and CXCR3 in cancer. AB - Chemokines comprise a superfamily of at least 46 cytokines that were initially described based on their ability to bind to 18 to 22 G protein-coupled receptors to induce the directed migration of leukocytes to sites of inflammation or injury. In addition to mediating cellular migration, chemokine/chemokine receptor pairs have been shown to affect many cellular functions, including survival, adhesion, invasion, and proliferation, and to regulate circulating chemokine levels. Most malignancies also express one or more chemokine receptors. Early studies established a role for CXCR4 and CXCR7 in mediating breast cancer metastasis, but other chemokine receptors, including CXCR3, now are implicated in several malignancies as biomarkers of tumor behavior as well as potential therapeutic targets. This review summarizes our current understanding regarding the contribution of CXCR4 and CXCR3 to tumor behavior and how receptor expression is regulated, transduces intracellular signals, and contributes at the molecular level to tumor behavior. It also describes recent therapeutic approaches that target these receptors or their ligands. PMID- 19216845 TI - Sarcomas of the head and neck region. AB - Sarcomas of the head and neck are a heterogeneous group of rare malignancies with highly variable clinical presentation, pathologic classification, and biologic behavior. Managing patients with sarcoma is a formidable challenge for surgical, medical, and radiation oncologists. This article reviews current approaches in patient evaluation, staging, prognosis, and treatment, with emphasis on prolonged survival and quality of life. PMID- 19216846 TI - Quality of life of oropharyngeal cancer patients treated with brachytherapy. AB - Brachytherapy (BT) is a highly conformal (accurate clinical target volume delineation, no planning target volume margin) radiotherapy technique; the radioactive source, guided by afterloading catheters, is implanted into the heart of the tumor. The localized high dose of radiation enables high tumor control rates and, because of rapid dose fall-off, sparing of the adjacent normal tissues. At the Erasmus Medical Center, excellent results were observed: 5-year local regional control of 84%, 5-year disease-free survival of 59%, and 5-year overall survival of 64%. Therefore, in the case of moderately sized tumors, for well-trained, skillful physicians, BT is the therapy of choice (if technically feasible). However, side effects are not totally negligible, partly because of the cumulative dose of BT and the first series of 46/2 Gy. However, patients treated with BT still have a better swallowing-related quality of life, which might improve further if summation of BT and the first series of 46/2 Gy, as well as autocontouring of the neck levels, are realized. So far, there is no significant relationship between the -quality index of the BT implants and local control/overall survival and/or quality of life. PMID- 19216847 TI - Surgical salvage of cancer of the oropharynx after chemoradiation. AB - Advanced oropharyngeal squamous cell carcinoma is treated primarily with chemoradiation, with the goal of excellent disease control and preservation of swallowing and articulation functions of the oropharynx. Disease control rates generally are excellent; however, a significant number of patients do not achieve locoregional control of disease. The importance of human papillomavirus expression in predicting successful tumor response, locoregional control, and survival following chemoradiation is increasingly confirmed. Emerging is a clinical profile, viral expression, and genetic expression pattern that can predict success of chemoradiation and indicate which patients are at higher risk for locoregional failure necessitating surgical salvage. Successful surgical salvage depends on restaging at the time of recurrence and the time interval from chemoradiation to recurrence. Although surgical morbidity and mortality remain a challenge in patients undergoing salvage surgery of the oropharynx after radiation failure with or without chemotherapy, they may be mitigated by liberal application of regional and microvascular free flap reconstruction techniques. PMID- 19216849 TI - [Analysis of toxicity of traditional Chinese herbal medicine and its connotation]. AB - Based on traditional Chinese medicine theory and clinical experience, traditional Chinese herbal drug toxicity has its own special connotation. From the perspective of history and logic, the different comprehension of toxicity between Western medicine and traditional Chinese medicine was discussed after retracing the meaning of "drug toxicity" in traditional Chinese medicine. The authors suggest that it's not feasible to study the Chinese medicine coping mechanically and applying indiscriminately the concept and the research idea about modern drug toxicity since there is different understanding of "drug toxicity" between traditional Chinese medicine and Western medicine. Many control elements are involved in the use of traditional Chinese herbal drugs, and Chinese drug components and actions are complex as compared with Western drugs. More and more drugs with toxicity will be found due to the relativity of drug toxicity. Currently, the study of Chinese drug toxicity should pay more attention to the relation between the toxicity and Chinese drug nature, compatibility and the corresponding disease or syndrome pattern after making definition of Chinese drug toxicity and its connotation. PMID- 19216848 TI - Immunotherapy of head and neck cancer using tumor antigen-specific monoclonal antibodies. AB - Monoclonal antibodies (mAbs) are now commonly used therapeutic agents in cancer patients. Since US Food and Drug Administration approval of cetuximab for head and neck squamous cell carcinoma, it has been used increasingly in this disease. Several other mAbs also are in development or in clinical -trials. Recently, evidence has accumulated that mAbs induce activation of cellular immunity, including natural killer and T cells and that this may contribute to clinical response. mAbs have been shown to mediate antibody-dependent cellular cytotoxicity, complement-dependent lysis, and activation of tumor antigen specific T cells. Various patient and tumor factors, such as polymorphisms in Fcgamma receptors expressed by immune cells, activity of T-regulatory cells, and tumor escape through downregulation of antigen-processing machinery in tumor cells, are likely to modulate the immune activation mediated by therapeutic mAbs. Understanding the interplay of these factors is likely to improve the selection of the most appropriate candidates for mAb-based immunotherapy, prediction of clinical response, and our understanding of mechanisms of tumor escape from therapeutic mAbs. PMID- 19216850 TI - [Therapy for clearing heat and resolving phlegm in treatment of systemic inflammatory response syndrome in acute deterioration stage of chronic obstructive pulmonary disease: a randomized controlled trial]. AB - OBJECTIVE: To investigate the effects of traditional Chinese medicine (TCM) therapy for clearing heat and resolving phlegm on systemic inflammatory response syndrome (SIRS) in acute deterioration stage of chronic obstructive pulmonary disease (COPD) and the serum procalcitonin (PCT) level. METHODS: One hundred patients suffering from SIRS in acute deterioration stage of COPD were included and randomly divided into treatment group (50 cases) and control group (50 cases). Patients in the treatment group were treated with routine Western medicine and Chinese herbal medicine for clearing away heat and eliminating phlegm, while patients in the control group were treated with routine Western medicine only. Patients in the two groups were treated for one week. The serum PCT levels in the two groups were measured at days 1, 3, and 7, and clinical symptom scores were recorded at days 1 and 7 in hospital to determine the effects. RESULTS: Clinical symptom scores in the two groups decreased after the treatment (P<0.05), and the symptom score in the treatment group was lower than that in the control group (P<0.05). The effect of integrated traditional Chinese and Western medicine treatment was obviously better than that of the Western medicine treatment when improving rates in clinical symptom score were divided into over 70% and less than 70% (P<0.05). Serum PCT concentration was in direct proportion to clinical symptom score (P<0.05). The patients with higher clinical symptom score were in more serious pathogenetic condition. There were no significant differences in the serum PCT concentrations among days 1, 3 and 7 in hospital. CONCLUSION: The effect of TCM therapy for clearing heat and removing phlegm combined with Western medicine treatment is better than that of the Western medicine treatment. TCM therapy has a satisfied effect in improving clinical symptoms. The serum PCT concentration is correlated with the severity of SIRS in acute deterioration stage of COPD. PMID- 19216851 TI - [Application of magnetic resonance imaging in evaluating the effects of manipulation on knee osteoarthritis]. AB - OBJECTIVE: To observe and evaluate the effects of manipulation on knee osteoarthritis (KOA) using T2-mapping and magnetic resonance imaging (MRI)-based volume measurements. METHODS: Forty-five cases of KOA were involved retrospectively in the study, and the patients were composed of 9 males and 36 females with the mean age of (57.4+/-6.1) years. The cases were treated with manipulative therapy once or twice per week for 12 months. MRI of each knee was performed separately by using 1.5-T MRI equipment before and during the treatment. Average cartilage depth, MRI grading of cartilage defects, cartilage volume, average T2 values in patella cartilage and femoral condyle, and bone marrow edema area were detected respectively. RESULTS: The knee joint cartilage thickness from MRI began to increase after 6-month treatment (P<0.05). The cartilage volume increase was obviously observed after 12-month treatment, and there was a significant difference (F=14.64, P<0.01). MRI grading of cartilage defects decreased from IIIA to IIB after 3-month treatment (Z=17.96, P<0.05). The average T2 value in patella cartilage decreased after 9-month treatment (F=3.11, P<0.05), but there were no differences in cartilage from tibial plateau and femoral condyle compartments after the treatment. The bone marrow edema area in femoral condyle began to diminish at 3-month treatment (t=-4.53, P<0.01), and the bone marrow edema area in cartilage patella was diminished after 6-month treatment (t=-5.53, P<0.01). CONCLUSION: T2-mapping and cartilage volume measurement are suitable for evaluating the manipulative therapy on KOA. Traditional Chinese manipulation therapy is an effective method for KOA in the cartilage recovery. PMID- 19216852 TI - [A family survey of syndromes of traditional Chinese medicine in patients with beta-thalassemia]. AB - OBJECTIVE: To explore the relationship between syndromes of traditional Chinese medicine (TCM) and genetic background in patients with beta-thalassemia. METHODS: TCM syndromes were surveyed in the selected 78 patients with beta-thalassemia intermedia including 120 parents. The gene mutations were detected separately. The frequency and score of TCM syndromes between the offspring and their parents in different family types were analyzed, and the differences were compared. RESULTS: The 73 families were divided into two family types by hereditary characteristics. Family type one meant that genotypes of one of the parents were normal, while the offspring genotypes were heterozygous and were exactly the same as another parent. In the 22 families of type one, the heterozygous offspring manifested 6 high-frequency symptoms and signs such as spontaneous perspiration, dry mouth and dry throat, pale or sallow complexion, tidal fever and night sweating, lassitude and pale fingernails. The heterozygous parents manifested 5 high-frequency symptoms and signs such as lassitude in loins and knees, dizziness, aversion to cold and cold limbs, tinnitus, dry mouth and dry throat. The normal parents manifested 3 high-frequency symptoms and signs such as lassitude in loins and knees, dizziness, and spontaneous perspiration. TCM syndrome score in the heterozygous offspring was higher than that in the heterozygous and normal parents, but there was no significant difference (P>0.05). Family type two meant that genotypes of both parents were heterozygous, while the offspring genotypes were heterogenic duplex heterozygotes. In the 51 families of type two, the offspring manifested 9 high-frequency symptoms and signs such as pale or sallow complexion, spontaneous perspiration, dry mouth and dry throat, pale fingernails, tidal fever and night sweating, lassitude, frequent attack of common cold, dysphoria with feverish sensation in chest, and yellow discoloration of the skin and sclera. The parents manifested 3 high-frequency symptoms and signs such as lassitude in loins and knees, dizziness, aversion to cold and cold limbs. TCM syndrome score in the offspring was significant higher than that in the parents (P<0.01). CONCLUSION: In the two family types, TCM syndrome in the offspring is of yin-blood deficiency, while the syndrome of the parents is of kidney deficiency. The differences of TCM syndromes between the offspring and the parents may have some relations to the type of mutant genes and genetically modified ingredients. This research provides scientific evidence to TCM syndrome differentiation treatment of thalassemia. PMID- 19216853 TI - [Effects of a formula of components from Shengmai Powder on glucocorticoid receptor in rats after thermal injury]. AB - OBJECTIVE: To investigate the effects of a formula of components from Shengmai Powder, a compound traditional Chinese herbal medicine, on glucocorticoid receptor (GR) in rats after thermal injury. METHODS: A total of 32 male SD rats were randomly assigned into normal control group, untreated group, ginsenosides group and components group, with 8 rats in each group. Rats in the normal control group were intragastrically administered with normal saline (NS) at room temperature once daily. Rats in the untreated group were treated with NS before thermal injury, and rats in the components group and ginsenosides group were once daily treated with a mixture of aqueous extracts of Ophiopogonis Japoni, Fructus schizandrae Chinensis and ginsenosides and ginsenosides respectively. Rats were administered for one week. After the last administration, rats in the untreated group and treated groups underwent thermal injury for one hour, and then were sacrificed immediately by decapitation. Blood serum was collected, and the serum corticosterone (CS) and adrenocorticotropic hormone (ACTH) levels were determined with enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) respectively. The liver, lung and kidney homogenates were used to determine the GR binding capacity by radioligand receptor binding assay. The results were analyzed by one point analysis. RESULTS: GR binding capacities in liver, lung and kidney cytosols in the untreated group were obviously lower than those in the normal control group (P<0.01). The GR binding capacities in the liver and lung cytosols in the components group and the ginsenosides group were significantly higher than those in the untreated group (P<0.01), but no significant difference was found in kidney cytosol. Compared with the ginsenosides group, GR binding capacity in liver cytosol in the components group was increased (P<0.01), but there were no noticeable differences when compared with the GR binding capacities in lung and kidney cytosols. Serum CS and ACTH levels of the normal rats were (66+/-16)microg/L and (59+/-18) ng/L respectively. There were significant differences in CS and ACTH levels between the normal control group and the other groups (P<0.01), in which the serum CS and ACTH levels were (113+/-33)microg/L and (125+/-20) ng/L, (123+/-26) microg/L and (110+/-30) ng/L and (118+/-17) microg/L and (115+/-35) ng/L respectively. But there was no significant difference between the untreated group and the other treated groups. CONCLUSION: The formula of components from Shengmai Powder can enhance the effect of ginsenosides in up-regulating GR in rats after thermal injury. PMID- 19216854 TI - [Effects of Feiyanning formula on expressions of mRNAs and proteins of CXCL12 and CXCR4 in mice with Lewis tumors]. AB - OBJECTIVE: To observe the effects of Feiyanning (FYN) formula, a compound traditional Chinese herbal medicine, on the expressions of CXCL12 and CXC chemokine receptor 4 (CXCR4) mRNAs and proteins in Lewis tumors in C57 mice. METHODS: A total of 24 C57BL-6 mice were used in this study. Lewis cells were subcutaneously injected to the right armpit to establish the tumor-bearing model. The C57 mice bearing Lewis tumor were randomly divided into untreated group, chemotherapy group, FYN group and chemotherapy plus FYN group. Reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry were employed to measure the expressions of CXCL12 and CXCR4 mRNAs and proteins in Lewis tumors in C57 mice respectively. RESULTS: The rates of lung cancer metastasis in the FYN group and the chemotherapy plus FYN group were markedly lower than that in the untreated group or the chemotherapy group. The expressions of CXCL12 and CXCR4 mRNAs and proteins appeared in tumor tissue in the untreated group. Although there was no significant difference in the expressions of mRNA and protein of CXCL12 among all groups (P>0.05), the expressions of mRNA and protein of CXCR4 in the untreated group and the chemotherapy group were markedly lower than those in the FYN group and the chemotherapy plus FYN group (P<0.05). CONCLUSION: FYN can inhibit the metastasis of Lewis tumor loaded in C57 mice. Its mechanisms may be related to its down-regulation of the expression of chemokine receptor CXCR4, and affecting the role of CXCL12-CXCR4 biological axis. PMID- 19216855 TI - [Effects of salidroside-pretreatment on neuroethology of rats after global cerebral ischemia-reperfusion]. AB - OBJECTIVE: To study the effects of salidroside-pretreatment on changes of neuroethology in rats with global cerebral ischemia-reperfusion injury so as to investigate its probable mechanism. METHODS: Sixty SD male rats were randomly divided into sham-operated group, untreated group and salidroside-pretreated group. The rats in salidroside-pretreated group were intraperitoneally administered with salidroside for seven days. The dose of salidroside was 12 mg/(kg.d). Thirty minutes after the last administration, the acute global cerebral ischemia-reperfusion in rats of the untreated group and the salidroside pretreated group was induced by using the modified Pulsinelli's 4-vessel occlusion method. Five rats in each group were killed to obtain their brains 24 hours after reperfusion. The water content in the right brain was measured by calculating the ratio of dry weight to wet weight of the right brain. Activity of superoxide dismutase (SOD) and content of malondialdehyde (MDA) in hippocampus of the rats were measured. Then neurological severity scores (NSSs) of the other 15 rats in each group were observed respectively before and 6, 12, 24, 48 and 96 h after reperfusion. At the fifth day after reperfusion, the test of Morris water maze was carried out to examine the memories and learning abilities of the rats. RESULTS: The content of MDA, the activity of SOD, the NSS, the mean incubation period and the ratio of time in the second quadrant in the untreated group were significant different from those in the sham-operated group (P<0.05). Compared with the untreated group, the brain water content, the content of MDA and the NSS degraded, and the mean incubation period shortened in salidroside-pretreated group. The activity of SOD and the ratio of residence time in the second quadrant increased in salidroside-pretreated group as compared with the untreated group (P<0.05). CONCLUSION: Salidroside can reduce the degree of cerebral edema of rats with global cerebral ischemia-reperfusion injury, relieve the metabolism abnormity of free radical and improve the function of cognition. PMID- 19216856 TI - [Effects of Qingkailing effective components on nuclear factor-kappa B in an ischemia-reperfusion injury model of rat brain microvascular endothelial cells in vitro]. AB - OBJECTIVE: To establish an ischemia-reperfusion injury model of rat cerebral microvascular endothelial cells (MVECs) in vitro, and to explore the relationship between nuclear factor-kappa B (NF-kappaB) and the protective effects of Qingkailing effective components (hyocholic acid, taurocholic acid, baicalin, jasminoidin, Pinctada martensii) on MVECs. METHODS: Brain MVECs of male rats were digested with trypsin and subcultured, then the content of MVECs was adjusted to 1x10 (5)/mL and the MVECs were divided into normal control group, untreated group, hyocholic acid group, taurocholic acid group, baicalin group, jasminoidin group, Pinctada martensii group and nimodipine group, with six holes in each group. Except for the normal control group, the MVECs in the other groups were exposed in oxygen and glucose deprivation (OGD) circumstance in vitro to simulate ischemia-reperfusion injury. Immunocytochemical staining and image analysis system were used to observe the expression of NF-kappaB protein. RESULTS: Under a light microscope, the nuclei of MVECs in the normal control group were blank. Staining intensity of NF-kappaB protein in the nucleus in the untreated group was much deeper than that in the endochylema, with NF-kappaB shifted to nucleus after activation; a small quantity of NF-kappaB protein were expressed in the border of nucleus next to endochylema in groups of Qingkailing effective components, and the NF-kappaB protein expression was weaker than that in the untreated group. With the image analysis, we found that transmittance of nucleus and endochylema in the untreated group was significantly lower than that in the normal control group (P<0.01). Transmittance of nucleus and endochylema in the treated groups was higher than that in the untreated group (P<0.05, P<0.01). CONCLUSION: Qingkailing effective components have significant effect in inhibiting NF-kappaB protein transferring from endochylema to nucleus in vitro. PMID- 19216857 TI - [Effects of salvianolate on cardiomyocytes apoptosis and heart function in a swine model of acute myocardial infarction]. AB - OBJECTIVE: To observe the effects of salvianolate on cardiomyocytes apoptosis and heart function in a swine model of acute myocardial infarction (AMI). METHODS: A total of 21 young swine were randomly divided into untreated group, low-dose salvianolate (LS) group and high-dose salvianolate (HS) group (7 in each group). AMI was induced by ligaturing left anterior descending coronary artery. After the operation, 400 or 200 mg salvianolate dissolved in 250 mL 5% glucose saline was administered by intravenous drip to swine in the HS group and the LS group respectively for 7 days. The swine in the untreated group were administered with 250 mL 5% glucose saline. The left ventricular ejection fraction (LVEF) and myocardial perfusion were measured by gated myocardial perfusion imaging at the end of the 4th week after operation. And myocardial apoptosis was detected by TdT mediated dUTP-biotin nick end labeling (TUNEL) method. Apoptosis index was calculated under an optical microscope. RESULTS: Myocardial apoptotic index in the edge of myocardial infarction was decreased in the HS group and the LS group, which in the HS group was lower than that in the untreated group (P<0.05). Radioactive defect regions were found by gated myocardial perfusion imaging in all the three groups, but those in the HS group and the LS group were less than those in the untreated group (P<0.05). And the levels of myocardial perfusion and LVEF in the HS group were significantly higher than those in the untreated group (P<0.05). CONCLUSION: Salvianolate administered by intravenous drip can inhibit the cardiomyocytes apoptosis and improve the function of heart after AMI in swine. PMID- 19216858 TI - Salvianolic acid B in vitro inhibited matrix metalloproteinases-1, -2, and -9 activities. AB - OBJECTIVE: Matrix metalloproteinases (MMPs) are a family of zinc-dependent endopeptidases, which as a group can degrade essentially all extracellular matrix components. The proteolytic property of the MMPs is important during wound healing to remove debris and facilitate cell migration. Targeting towards the decreased MMPs activities is a new treatment strategy for healing chronic wounds. Salvia miltiorrhiza is a popular Chinese herb that could promote chronic ulcers healing for topical use. Salvianolic acid B (Sal B) is the most abundant bioactive component in Salvia miltiorrhiza. The research was designed to explore the inhibitory effects of Sal B on MMP-1, MMP-2 and MMP-9 activities. METHODS: Pure human interstitial collagenase (MMP-1) or gelatinase A (MMP-2) was activated by p-aminophenylmercuric acetate (APMA), and was incubated with Sal B for 1 h. The activities were observed by quenched fluorescent substrate. Gelatinase B (MMP 9) is rich in polymorphonuclear neutrophils (PMN), so the rat PMN was used as a source of MMP-9 for MMPs activity assays. In vitro MMP-9 from rats' PMN lysate was incubated with Sal B for 1 h, and its activity was tested by gelatin zymography. RESULTS: Sal B dose-dependently inhibited the human MMP-1 and MMP-2 activities in the range of 0.002 4 to 0.3 g/L, with 50% inhibiting concentration (IC(50)) of (0.090<0.015) g/L and (0.080<0.005) g/L respectively. In the range of 0.003 to 0.3 g/L, Sal B could inhibit the MMP-9 activity (P<0.01). CONCLUSION: The broad-spectrum inhibitory effects of Sal B on MMPs may reveal one of the mechanisms for the effects of Salvia miltiorrhiza on chronic wounds. PMID- 19216859 TI - [Effects of Salviae miltiorrhizae and salvianolic acid B on microcirculation of liver in mice with portal hypertension]. AB - OBJECTIVE: To observe the effects of Salviae miltiorrhizae and its component, salvianolic acid B (SA-B), on the microcirculation of liver in mice with portal hypertension induced by endothelin-1 (ET-1). METHODS: Eighty-four Kunming mice were randomly divided into 7 groups: untreated group, endothelin A receptor (ETAR) blocker group, Astragali mongolici group, Astragalus polysaccharides (APS) group, Corydalis Yanhusuo group, Salviae miltiorrhizae group and SA-B group. There were 12 mice in each group. Mice were pretreated with a corresponding equivalent volume of drug or distilled water for 3 days, and then the portal hypertension in mice was induced by continuous injection of ET-1 into coccygeal vein using a micro-injection pump. Six mice in each group were used to observe the average liver blood flow volume by laser-Doppler flow instrument before and after injection of ET-1, and the other six rats were used to observe the hepatic microcirculation velocity in vivo by an inverted microscope. RESULTS: The average blood flow of liver in mice decreased in each group after ET-1 injection. But the changes of average blood flow in the SA-B group and the ETAR blocker group were less than that in the untreated group (P<0.01). The changes of average blood volume in the Astragali mongolici group and the APS group were similar to that in the untreated group, but more than that in the SA-B group after injection of ET 1. The change of average blood flow in the SA-B group showed no significant difference when compared with the ETAR blocker group. The microcirculatory flow velocity in liver also decreased in each group after ET-1 injection. But the changes of microcirculatory flow velocity in the SA-B group and the ETAR blocker group were less than that in the untreated group (P<0.05, P<0.01). There were no significant differences in the changes of microcirculatory flow velocity among the Salviae miltiorrhizae group, the SA-B group and the ETAR blocker group. CONCLUSION: Salviae miltiorrhizae and SA-B can decrease the average blood flow and microcirculatory flow velocity in liver in mice with portal hypertension, which may be one of the mechanisms of Salviae miltiorrhizae and SA-B in decreasing portal hypertension. PMID- 19216860 TI - [Treatment of adjuvant arthritis in rats with Chinese herbal fumigation: efficacy and mechanism]. AB - OBJECTIVE: To observe the efficacy of fumigation with Fumigant I formula, a compound traditional Chinese herbal medicine, in treating adjuvant arthritis (AA) in rats and to explore its anti-inflammation mechanism. METHODS: Fifty male Wistar rats were randomly divided into normal control group, untreated group, distilled-water fumigation group, and low- and high-dose herb fumigation groups. Except for the normal control group, the other rats were used to induce AA by complete Freund's adjuvant injection. After treatments, the degree of foot swelling, the pathological changes and expressions of the inflammatory cytokines including interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and intercellular adhesion molecule-1 (ICAM-1) in the ankle joint were detected. The contents of these cytokines in the serum were also observed. RESULTS: In two Chinese herbal fumigation groups, as compared with the untreated group, foot swelling was obviously reduced (P<0.05); the pathological changes of the ankle joint, such as inflammatory cell infiltration, fibrous tissue and synovial cell proliferation in the ankle joint were obviously improved (P<0.05, P<0.01); the positive expressions of IL-1beta, TNF-alpha, ICAM-1 in the ankle joint were significantly down-regulated (P<0.05); and the contents of these inflammatory cytokines in the serum were obviously decreased (P<0.05). The treatment effect in high-dose herb fumigation group was better than that in low-dose herb fumigation group and that in distilled-water fumigation group (P<0.05, P<0.01). CONCLUSION: Fumigation with Chinese herbal medicine has a positive therapeutic effect on AA rats, and its anti-inflammatory mechanism may be related with inhibiting inflammatory cytokines IL-1beta, TNF-alpha and ICAM-1. PMID- 19216861 TI - [Preventive and therapeutic effects of Yiqi Huayu Recipe on degeneration of articular cartilage in rats with osteoarthritis]. AB - OBJECTIVE: To study the efficacy and possible mechanism of Yiqi Huayu Recipe (YQHYR), a compound traditional Chinese herbal medicine, in preventing and treating degeneration of the articular cartilage in rats with osteoarthritis (OA). METHODS: A total of 90 one-month-old SD rats were randomly divided into normal control group, untreated group and YQHYR group, with 30 rats in each group. The osteoarthritis was induced by shoulder disarticulation plus upright posture in rats. The rats in YQHYR group were treated with YQHYR at 5-, 7- and 9 month old (4, 6 and 8 months after the surgery) for one month. Ten rats in each subgroup were sacrificed at 6-, 8- and 10-month old (5, 7 and 9 months after the surgery) respectively and the knee joint samples were harvested for detection. Safranin-O/fast green staining was performed to examine the morphology of articular cartilage. The expressions of type II collagen (Col2A1), aggrecan-1 (Agc1), matrix metalloproteinase-13 (MMP-13) and tissue inhibitor of metalloproteinase-1 (TIMP-1) mRNAs were evaluated by real-time fluorescent quantitation polymerase chain reaction. RESULTS: Histological analysis showed that the structure of articular cartilage was seriously destroyed in rats in the untreated group. On the contrary, the degenerative changes of the articular cartilage in the YQHYR group were dropped off. Real-time fluorescent quantitation polymerase chain reaction showed that expressions of Agc1, TIMP-1 and Col2A1 mRNAs were up-regulated in 6- and 10-month-old rats in the YQHYR group as compared with those in the untreated group (P<0.01, P<0.05), but there was no significant difference in expression of MMP-13 mRNA between the YQHYR group and the untreated group. The expression of Agc1 mRNA was up-regulated and the expression of MMP-13 mRNA was down-regulated in the 8-month-old rats in YQHYR group as compared with those in the untreated group (P<0.01). CONCLUSION: YQHYR can promote the synthesizing of aggrecan and type II collagen in chondrocytes and delay articular cartilage degradation in OA rats. PMID- 19216862 TI - [Clinical application of Dahuang Zhechong Pill in gynecology]. PMID- 19216863 TI - [Syndrome differentiation and treatment of Taiyang disease in Shanghan Lun]. AB - OBJECTIVE: To explore the laws in syndrome differentiation of Taiyang disease and the prescriptions and herbs used in its treatment in Shanghan Lun (Treatise on Febrile Diseases). METHODS: The occurrence rates of main syndromes of Taiyang disease, and the usage frequency of the prescriptions and herbs in its treatment were calculated, and the laws in syndrome differentiation and herbal medication were analyzed by hierarchical clustering analysis. RESULTS: Fever and aversion to cold were found to be the main symptoms of Taiyang disease and usually accompanied with headache, absent sweating, neck stiff, floating and tight pulse, and body ache. The accompanying or aggravated symptoms could be classified into lung system syndrome with the manifestations of sweating and asthma, spleen deficiency syndrome with the manifestations of gastric fullness and diarrhea, stomach syndrome with vomit and constipation. Guizhi decoction was the main prescription used in the treatment of Taiyang disease. Mahuang (Herba Ephedrae) matching Xingren (Semen Armeniacae), Dahuang (Radix et Rhizoma Rhei) matching Mangxiao (Natrii Sulfas), Baizhu (Rhizoma Atractylodes Macrocephalae) matching Fuling (Poria), Fuzi (Radix Aconiti Lateralis) matching Ganjiang (Rhizoma Zingiberis), Chaihu (Radix Bupleuri) matching Huangqin (Radix Scutellariae), Banxia (Rhizoma Pinelliae), and Renshen (Radix Ginseng) as the main compatibilities were used in treating lung diseases, stomach diseases, spleen deficiency diseases, kidney-deficiency diseases and Shaoyang diseases respectively. CONCLUSION: Exterior syndrome, as a common syndrome in Taiyang disease, is usually treated with Guizhi decoction. The change in syndromes from upper-energizer to lower-energizer in exterior disease can be found from the change of symptoms and the use of herbs. And the development from defending stage to qi stage in exterior disease can be found in the use of prescriptions in Shanghan Lun. PMID- 19216864 TI - [A review of studies on active agents and pharmacology of Thyme]. PMID- 19216865 TI - [How to express and describe quantitative data: part one]. PMID- 19216866 TI - Comparative study on WHO Western Pacific Region and World Federation of Chinese Medicine Societies international standard terminologies on traditional medicine: an analysis of the Five Sensory Organs. PMID- 19216867 TI - [Influential factors of translation unit in traditional Chinese medicine]. PMID- 19216868 TI - [Traditional Chinese medicine in France: French Federation of Traditional Chinese Medicine]. PMID- 19216869 TI - Artificial neural networks in foodstuff analyses: Trends and perspectives A review. AB - Artificial neural networks are a family of non-linear computational methods, loosely inspired by the human brain, that have found application in an increasing number of fields of analytical chemistry and specifically of food control. In this review, the main neural network architectures are described and examples of their application to solve food analytical problems are presented, together with some considerations about their uses and misuses. PMID- 19216870 TI - Field-flow fractionation in bioanalysis: A review of recent trends. AB - Field-flow fractionation (FFF) is a mature technique in bioanalysis, and the number of applications to proteins and protein complexes, viruses, derivatized nano- and micronsized beads, sub-cellular units, and whole cell separation is constantly increasing. This can be ascribed to the non-invasivity of FFF when directly applied to biosamples. FFF is carried out in an open-channel structure by a flow stream of a mobile phase of any composition, and it is solely based on the interaction of the analytes with a perpendicularly applied field. For these reasons, fractionation is developed without surface interaction of the analyte with packing or gel media and without using degrading mobile phases. The fractionation device can be also easily sterilized, and analytes can be maintained under a bio-friendly environment. This allows to maintain native conditions of the sample in solution. In this review, FFF principles are briefly described, and some pioneering developments and applications in the bioanalytical field are tabled before detailed report of most recent FFF applications obtained also with the hyphenation of FFF with highly specific, sensitive characterization methods. Special focus is finally given to the emerging use of FFF as a pre analytical step for mass-based identification and characterization of proteins and protein complexes in proteomics. PMID- 19216871 TI - Factorial-based response-surface modeling with confidence intervals for optimizing thermal-optical transmission analysis of atmospheric black carbon. AB - Thermal-optical transmission (TOT) analysis measures black carbon (BC) in atmospheric aerosol on a fibrous filter. The method pyrolyzes organic carbon (OC) and employs laser light absorption to distinguish BC from the pyrolyzed OC; however, the instrument does not necessarily separate the two physically. In addition, a comprehensive temperature protocol for the analysis based on the Beer Lambert Law remains elusive. Here, empirical response-surface modeling was used to show how the temperature protocol in TOT analysis can be modified to distinguish pyrolyzed OC from BC based on the Beer-Lambert Law. We determined the apparent specific absorption cross sections for pyrolyzed OC (sigma(Char)) and BC (sigma(BC)), which accounted for individual absorption enhancement effects within the filter. Response-surface models of these cross sections were derived from a three-factor central-composite factorial experimental design: temperature and duration of the high-temperature step in the helium phase, and the heating increase in the helium-oxygen phase. The response surface for sigma(BC), which varied with instrument conditions, revealed a ridge indicating the correct conditions for OC pyrolysis in helium. The intersection of the sigma(BC) and sigma(Char) surfaces indicated the conditions where the cross sections were equivalent, satisfying an important assumption upon which the method relies. 95% confidence interval surfaces defined a confidence region for a range of pyrolysis conditions. Analyses of wintertime samples from Seattle, WA revealed a temperature between 830 degrees C and 850 degrees C as most suitable for the helium high-temperature step lasting 150s. However, a temperature as low as 750 degrees C could not be rejected statistically. PMID- 19216872 TI - A spectral similarity measure using Bayesian statistics. AB - A spectral similarity measure was developed that can differentiate subtle differences between two spectra. The spectra are digitalized into a vector. The difference between the two spectra is defined by a difference vector, which is one spectrum minus the other. The spectral similarity measure is transformed into a hypothesis test of the similarities and differences between the two spectra. The scalar mean of the difference vector is used as the statistical variable for the hypothesis test. A threshold for the hypothesis that the spectra are different was proposed. The Bayesian prior odds ratio was estimated from multiple spectra of the same sample. The posterior odds ratio was used to quantity the spectral similarity measure of the two spectra. Diffuse reflectance near-infrared spectra of tobacco samples of two formulations were used to demonstrate this method. The results show that this new method can detect subtle differences between the spectra. PMID- 19216873 TI - Stir bar sorptive extraction-gas chromatography-mass spectrometry analysis of tetramethylene disulfotetramine in food: Method development and comparison to solid-phase microextraction. AB - A stir bar sorptive extraction-gas chromatography-mass spectrometry (SBSE-GC-MS) method for the determination of tetramethylene disulfotetramine is presented. The limits of detection (LOD) of the optimized method was 0.2ngg(-1) for extractions from water and 0.3-2.1ngg(-1) for extractions from foods. Recovery was highly matrix dependent (36-130%) and quantification required standard addition calibrations. Standard addition calibration lines had high linearity (R(2)>0.97) and replicate extractions had good reproducibility (R.S.D.=4.4-9.8%). A comparison of the SBSE method and a previously developed headspace (HS)-solid phase microextraction (SPME) method was performed. Generally, SBSE provided higher sensitivity with decreased analysis time. PMID- 19216874 TI - Headspace solid phase microextraction and gas chromatography-quadrupole mass spectrometry methodology for analysis of volatile compounds of marine salt as potential origin biomarkers. AB - The establishment of geographic origin chemical biomarkers for the marine salt might represent an important improvement to its valorisation. Volatile compounds of marine salt, although never studied, are potential candidates. Thus, the purpose of this work was the development of a headspace solid phase microextraction (SPME) combined with gas chromatography-quadrupole mass spectrometry (HS-SPME/GC-qMS) methodology to study the volatile composition of marine salt. A 65mum carbowax/divinylbenzene SPME coating fibre was used. Three SPME parameters were optimised: extraction temperature, sample quantity, and presentation mode. An extraction temperature of 60 degrees C and 16g of marine salt in a 120mL glass vial were selected. The study of the effect of sample presentation mode showed that the analysis of an aqueous solution saturated with marine salt allowed higher extraction efficiency than the direct analysis of salt crystals. The dissolution of the salt in water and the consequent effect of salting-out promote the release of the volatile compounds to the headspace, enhancing the sensitivity of SPME for the marine salt volatiles. The optimised methodology was applied to real matrices of marine salt from different geographical origins (Portugal, France, and Cape Verde). The marine salt samples contain ca. 40 volatile compounds, distributed by the chemical groups of hydrocarbons, alcohols, phenols, aldehydes, ketones, esters, terpenoids, and norisoprenoids. These compounds seem to arise from three main sources: algae, surrounding bacterial community, and environment pollution. Since these volatile compounds can provide information about the geographic origin and saltpans environment, this study shows that they can be used as chemical biomarkers of marine salt. PMID- 19216875 TI - Characterization of the quantitative relationship between signal-to-noise (S/N) ratio and sample amount on-target by MALDI-TOF MS: Determination of chondroitin sulfate subsequent to enzymatic digestion. AB - Chondroitin sulfate (CS) is an important glycosaminoglycan of the extracellular matrix and its quantitative detection is of interest in different pathologies. Although there are already methods of quantitative CS determination, many of them are laborious, require time-consuming sample workup and/or suffer from low sensitivity. It will be shown here that the CS content of biological samples can be easily assessed in the picomole range subsequent to enzymatic digestion. MALDI TOF MS (matrix-assisted laser desorption and ionization time-of-flight mass spectrometry) was used to determine the concentrations of the unsaturated disaccharide of CS obtained by enzymatic digestion of native CS with chondroitin ABC lyase. The signal-to-noise (S/N) ratio can be used as a quantitative measure: amounts of CS (measured as the disaccharide) down to at least 500fmol could be detected and there is a direct correlation between the S/N ratio and the amount of CS between about 2 and 200pmol although the curve per se is sigmoidal. The influence of different parameters such as the used matrix, the applied laser intensity and different methods of data analysis were also tested. Advantages and drawbacks of this approach are critically discussed. Finally, the method was validated by the determination of the CS content in samples of known concentration as well as in enzymatically digested bovine nasal cartilage and compared with two further established methods of CS determination (Carbazole and Alcian Blue method). PMID- 19216876 TI - A cataluminescence gas sensor for carbon tetrachloride based on nanosized ZnS. AB - A novel and sensitive gas sensor was proposed for the determination of carbon tetrachloride based on its cataluminescence (CTL) by oxidation in the air on the surface of nanosized ZnS. The luminescence characteristics and the optimal conditions were investigated in detail. Under the optimized conditions, the linear range of the CTL intensity versus the concentration of carbon tetrachloride was 0.4-114microg mL(-1), with a correlation coefficient (R) of 0.9986 and a limit of detection (S/N=3) of 0.2microg mL(-1). The relative standard deviation (R.S.D.) for 5.9microg mL(-1) carbon tetrachloride was 2.9% (n=5). There was no or weak response to common foreign substances including methanol, ethanol, benzene, acetone, formaldehyde, acetaldehyde, dichloromethane, xylene, ammonia and trichloromethane. There was no significant change of the catalytic activity of the sensor for 40h over 4 days, with a R.S.D. of less than 5% by collecting the CTL intensity once an hour. The proposed method was simple and sensitive, with a potential of detecting carbon tetrachloride in environment and industry grounds. The possible mechanism was also discussed briefly. PMID- 19216877 TI - Rapid and novel discrimination and quantification of oleanolic and ursolic acids in complex plant extracts using two-dimensional nuclear magnetic resonance spectroscopy-Comparison with HPLC methods. AB - A novel strategy for NMR analysis of mixtures of oleanolic and ursolic acids that occur in natural products is described. These important phytochemicals have similar structure and their discrimination and quantification is rather difficult. We report herein the combined use of proton-carbon heteronuclear single-quantum coherence ((1)H-(13)C HSQC) and proton-carbon heteronuclear multiple-bond correlation ((1)H-(13)C HMBC) NMR spectroscopy, in the identification and quantitation of oleanolic acid (OA) and ursolic acid (UA)in plant extracts of the Lamiaceae and Oleaceae family. The combination of (1)H (13)C HSQC and (1)H-(13)C HMBC techniques allows the connection of the proton and carbon-13 spins across the molecular backbone resulting in the identification and, thus, discrimination of oleanolic and ursolic acid without resorting to physicochemical separation of the components. The quantitative results provided by 2D (1)H-(13)C HSQC NMR data were obtained within a short period of time ( approximately 14min) and are in excellent agreement with those obtained by HPLC, which support the efficiency of the suggested methodology. PMID- 19216878 TI - Imaging fingerprinting of excitation emission matrices. AB - The spectral fingerprinting of the excitation emission matrix (EEM) of fluorescent substances is demonstrated using polychromatic light sources and tri chromatic image detectors. A model of the measured fingerprints explaining their features and classification performance, based on the polychromatic excitation of the indicators is proposed. Substantial amount of spectral information is retained in the fingerprints as corroborated by multivariate analysis and experimental conditions that favor such situation are identified. In average, for five different substances, the model shows a fitting goodness measured by the Pearson's r coefficient and the root mean square deviation of 0.8541 and 0.0247 respectively, while principal component classification patterns satisfactorily compare with the EEM spectroscopy classification and respectively explain 96% and 93% of the information in the fist two principal components. The measurements can be performed using regular computer screens as illumination and web cameras as detectors, which constitute ubiquitous and affordable platforms compatible with distributed evaluations, in contrast to regular instrumentation for EEM measurements. PMID- 19216879 TI - Determination of dextrin based on its self-aggregation by resonance light scattering technique. AB - A novel free-probe assay of dextrin was established based on the resonance light scattering (RLS) enhancement in aqueous solution due to the self-aggregation of dextrin. The RLS intensity was well proportional to the concentration of dextrin over the wide range 0.2-14microg mL(-1) and a detection limit 0.02microg mL(-1) was obtained in the optimum conditions. The effect factors such as pH, buffer medium, holding time, ionic strength and temperature were studied in detail. Little or no interference was presented in the detection when adding coexisting substances including various metal ions and some saccharine in the solution. The assay proposed owns the advantages of easy operation, rapidity, sensitivity and practicability. Three synthetic samples and three kinds of medicine samples were analyzed with satisfactory results. PMID- 19216880 TI - Development of a fluorescent chelating ligand for gallium ion having a quinazoline structure with two Schiff base moieties. AB - A novel chelating ligand, 2,4-[bis-(2,4-dihydroxybenzylidene)] dihydrazinoquinazoline (DBHQ), was synthesized, and the fluorescence characteristics of its complex with metal ions were investigated. Thirty-five different metal ions were tested for the emission of fluorescence in the presence of DBHQ in aqueous solutions in a pH range of 3.0-10.5 (at a difference of 0.5 for each metal). It was observed that DBHQ fluoresces intensely at 470nm with an excitation wavelength of 405nm in the presence of Ga(3+) or Al(3+) in the pH range 3.0-4.0. The other metal ions did not show fluorescence with DBHQ. Although the presence of Cu(2+), Co(2+) and Fe(3+) decreased the fluorescence intensity of DBHQ-Ga(3+), the addition of a fluoride ion (NaF) recovered the fluorescence by masking the interfering ions. In addition, the fluoride ions were found to enhance the sensitive determination of Ga(3+) because the fluorescence intensity of DBHQ-Ga(3+) was further increased approximately 2.5-fold in the presence of F( ) (phi=0.658) from that in the absence of F(-) (phi=0.401). The fluoride ions also masked the Al(3+) ions, which emit fluorescence on chelation with DBHQ. Therefore, a selective and sensitive detection of Ga(3+) was achieved by using DBHQ in the presence of F(-). The detection limit of Ga(3+) was approximately 50nmolL(-1) (3.5ppb). The proposed method was applicable to determine Ga(3+) in river water. PMID- 19216881 TI - Quantification approach for assessment of sparkling wine volatiles from different soils, ripening stages, and varieties by stir bar sorptive extraction with liquid desorption. AB - Stir bar sorptive extraction with liquid desorption followed by large volume injection coupled to gas chromatography-quadrupole mass spectrometry (SBSE-LD/LVI GC-qMS) was applied for the quantification of varietal and fermentative volatiles in sparkling wines. The analytical data were performed by using suitable standards of monoterpene hydrocarbons (alpha-pinene), monoterpenols (linalool), sesquiterpenoids (E,E-farnesol, Z-nerolidol, and guaiazulene), C(13) norisoprenoids (beta-ionone), aliphatic and aromatic alcohols (hexanol and 2 phenylethanol), and esters (hexyl acetate and ethyl decanoate) as model compounds. The wine volatiles were quantified using the structurally related standards. The methodology showed good linearity over the concentration range tested, with correlation coefficients ranging from 0.950 to 0.997, and a reproducibility of 9-18%. The SBSE-LD/LVI-GC-qMS methodology allowed, in a single run, the quantification of 71 wine volatiles that can be quantified accurately at levels lower than their respective olfactory thresholds. This methodology was used for assessment of sparkling wine volatiles from different soils, ripening stages, and varieties. The variety and soil influenced significantly the volatile composition of sparkling wines; lower effect was observed for the ripening stage of grapes picked up one week before or after the maturity state. PMID- 19216883 TI - Application of principal component analysis to the thermal characterization of silanized nanoparticles obtained at supercritical carbon dioxide conditions. AB - Samples resulting from reaction of TiO(2) with octyltriethoxysilane, developed using a supercritical carbon dioxide procedure, have been studied by infrared and Raman spectroscopies and thermogravimetric analysis. Different reaction conditions have been applied to the preparation of samples in order to study the influence of experimental factors on the sample properties. Vibrational techniques have first been used to verify the presence of silanized structures on the surface of TiO(2) through the detection of specific bands characteristic of the Si-O-Si cross-linking. Thermogravimetric profiles consisting of weight loss values as a function of temperature have been analyzed by principal component analysis to extract information about the characteristics of the linkage between silane and TiO(2) as well as the thermal stability of the prepared materials. The mathematical treatment of data has provided conclusions on the properties of the samples and analogies and differences with respect to the commercial material. PMID- 19216882 TI - Residue determination of glyphosate in environmental water samples with high performance liquid chromatography and UV detection after derivatization with 4 chloro-3,5-dinitrobenzotrifluoride. AB - A pre-column derivatization high-performance liquid chromatographic method for glyphosate analysis has been developed. Derivatization of glyphosate was performed with 4-chloro-3,5-dinitrobenzotrifluoride (CNBF). In pH 9.5 H(3)BO(3) Na(2)B(4)O(7) media, the reaction of glyphosate with CNBF completed at 60 degrees C for 30min. The labeled glyphosate was separated on a Kromasil C18 column (250mmx4.6mm, 5microm) at room temperature and UV detection was applied at 360nm. The separation of labeled glyphosate was achieved within 15min by gradient elution mode. Compared to other pre-column derivatization, this derivatization was performed more mildly, the derivative was more stable, and the detection limits of a few reagents were higher than CNBF, except 9-fluorenylmethyl chloroformate (FMOC-Cl) using fluorescence and mass spectrometry, however, this reagent avoid to be removed after derivatization like FMOC-Cl. The detection limit of glyphosate was 0.009mgL(-1) (S/N=3) without preconcentration and reach MRL, which is set at the level of 0.1mgL(-1) in China. The method linearity correlation coefficient was 0.9999, in concentrations ranging from 0.3 to 48.5mgL(-1). The proposed method has been applied to the quantitative determination of glyphosate in environmental water with recoveries of 91.80 100.20% and R.S.D. of 2.27-6.80, depending on the sample investigated. PMID- 19216884 TI - Addiction and suicidal behavior in acute psychiatric inpatients. AB - OBJECTIVE: This study aims to evaluate the relationship of alcohol/drug use and effect severities to the degree of suicidality in acutely admitted psychiatric patients. Both degree of substance dependency and degree of substance-induced syndrome were analyzed. In addition, length of stay, involuntary status, and against medical advice discharge status were determined as they related to these variables. METHODS: Structured clinical admissions and discharge ratings were gathered from 10,667 consecutive, single-case individual records, from an urban acute care county psychiatric hospital. RESULTS: Data indicate that of the most severely suicidal group, 56% had substance abuse or dependence, 40% were rated as having half or more of their admission syndrome substance induced, and most had nonpsychotic diagnoses. There was an inverse relationship between degree of substance problem and length of stay. Although these patients more commonly left against medical advice, and were readmitted more frequently, they were less likely to be involuntarily committed. CONCLUSIONS: A large, potentially lethal, and highly expensive subgroup of patients has been characterized, which might be called the "New Revolving Door acute psychiatric inpatient." This group, which uses the most expensive level of care in the mental health system but is substantially addiction related, poses special challenges for inpatient psychiatric units, addiction treatment providers, and health care planners. PMID- 19216886 TI - General health and quality-of-life measures in active, recent, and comorbid mental disorders: a population-based health 2000 study. AB - AIMS: We studied the impact of comorbidity and recency in psychiatric disorders on psychological well-being, perceived health, and quality of life and compared their effect with the effect of a chronic medical condition, type 2 diabetes mellitus. METHODS: Established instruments for psychological distress (12-item General Health Questionnaire [GHQ-12]), self-rated general health, and health related quality of life (EQ-5D and 15D) were administered for the participants of the nationwide Finnish Health 2000 survey. The diagnoses of depressive, anxiety, and alcohol use disorders and their unique comorbid combinations were based on the Munich version of the Composite International Diagnostic Interview (M-CIDI). RESULTS: People with comorbid anxiety and depressive disorders had the highest distress and lowest health and quality-of-life ratings on all scales. The effects of mental disorders on health-related quality of life and self-rated health were comparable to the effects of diabetes. Type 2 diabetes mellitus and alcohol use disorder were associated with minimal, although statistically significant, increase of psychological distress. Symptom recovery from an active depressive disorder associated with improved well-being on all measures, but residual ill being was also remarkably common among the partly or fully recovered in all disorder categories. CONCLUSIONS: Even in comparison with a chronic physical illness such as diabetes, comorbid psychiatric disorders have a high impact on psychological well-being, perceived health, and quality of life, when evaluated by simple, useful, and feasible self-rating scales. The scales used may be useful in monitoring the severity and course of psychiatric conditions. Recognizing and treating psychiatric comorbidity is important in improving the quality of life of psychiatric patients. PMID- 19216885 TI - Predictors of nonadherence among individuals with bipolar disorder receiving treatment in a community mental health clinic. AB - BACKGROUND: Subjective experience of illness is a critical component of treatment adherence in populations with bipolar disorder (BPD). This cross-sectional analysis examined clinical and subjective variables in relation to adherence in 140 individuals with BPD receiving treatment with mood-stabilizing medication. METHODS: Nonadherence was defined as missing 30% or more of medication on the Tablets Routine Questionnaire, a self-reported measure of medication treatment adherence. Adherent and nonadherent groups were compared on measures of attitudes toward illness and treatment including the Attitudes toward Mood Stabilizers Questionnaire, the Insight and Treatment Attitudes Questionnaire, the Rating of Medication Influences, and the Multidimensional Health Locus of Control Scale. RESULTS: Except for substance abuse comorbidity, adherent individuals (n = 113, 80.7%) did not differ from nonadherent individuals (n = 27, 19.3%) on clinical variables. However, nonadherent individuals had reduced insight into illness, more negative attitudes toward medications, fewer reasons for adherence, and more perceived reasons for nonadherence compared with adherent individuals. The strongest attitudinal predictors for nonadherence were difficulties with medication routines (odds ratio = 2.2) and negative attitudes toward drugs in general (odds ratio = 2.3). LIMITATIONS: Results interpretation is limited by cross-sectional design, self-report methodology, and sample size. CONCLUSIONS: Comorbid substance abuse, negative attitudes toward mood-stabilizing medication, and difficulty managing to take medication in the context of one's daily schedule are primary determinants of medication treatment adherence. A patient-centered collaborative model of care that addresses negative attitudes toward medication and difficulty coping with medication routines may be ideally suited to address individual adherence challenges. PMID- 19216887 TI - Risk factors associated with lifetime suicide attempts in bipolar I patients: findings from a French National Cohort. AB - Risk factors that may be associated with suicide attempts in bipolar disorder are still a matter of debate. We compared demographic, illness course, clinical, and temperamental features of suicide attempters vs those of nonattempters in a large sample of bipolar I patients admitted for an index manic episode. One thousand ninety patients (attempters = 382, nonattempters = 708) were included in the study. Multivariate analysis evidenced 8 risk factors associated with lifetime suicide attempts as follows: multiple hospitalizations, depressive or mixed polarity of first episode, presence of stressful life events before illness onset, younger age at onset, no free intervals between episodes, female sex, higher number of previous episodes, and cyclothymic temperament. These characteristics may help identify subjects at risk for suicide attempt throughout the course of bipolar disorder. We finally propose to integrate such characteristics into a stress-diathesis model of suicidal behavior, adapted to bipolar patients. PMID- 19216888 TI - Cumulative effect of multiple trauma on symptoms of posttraumatic stress disorder, anxiety, and depression in adolescents. AB - BACKGROUND: Recent literature has indicated that exposure to multiple traumatic events in adults is associated with high levels of posttraumatic stress disorder (PTSD), anxiety, and depression. Against the backdrop of stressful life events and childhood abuse and neglect, we investigated the cumulative effect of multiple trauma exposure on PTSD, anxiety, and depression in an adolescent sample. METHOD: One thousand one hundred forty 10th-grade learners from 9 Cape Town (South Africa) schools completed questionnaires on stressful life experiences; trauma exposure; and symptoms of anxiety, depression, and PTSD. Our population of interest for this study was adolescents between the ages of 14 and 18 years who had been exposed to serious, Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, qualifying traumatic events. The final sample size was thus 922. RESULTS: Rates of trauma exposure, PTSD, depression, and anxiety were high. Controlling for sex, stressful life experiences in the past year, and childhood adversity, we found an effect of cumulative trauma exposure effect on PTSD and depression, with an increase in the number of traumas linearly associated with an increase in symptoms of PTSD (F((4,912)) = 7.60, P < .001) and depression (F((4,912)) = 2.77, P < .05). We did not find a cumulative effect on anxiety. CONCLUSION: Our findings indicate that adolescents exposed to multiple traumas are more likely to experience more severe symptoms of PTSD and depression than those who experience a single event, with this effect independent of childhood adversity and everyday stressful life experiences. Exposure to multiple trauma, however, does not seem to be associated with more severe anxiety symptoms. PMID- 19216889 TI - A more proximal impact of dissociation than of trauma and posttraumatic stress disorder on schneiderian symptoms in patients diagnosed with schizophrenia. AB - BACKGROUND: Dissociation is often related to psychologic trauma and is also commonly correlated with posttraumatic stress disorder (PTSD). Schneiderian symptoms occur in dissociative disorder and PTSD, whereas dissociation and trauma are also frequent in patients with schizophrenia. Dissociation and schneiderian symptoms may reflect posttraumatic symptomatology in some patients diagnosed with schizophrenia who might not develop PTSD. METHOD: Seventy-one patients completed an interview assessing symptoms of schizophrenia, depression, dissociative disorder, and PTSD. Data were collected using the Positive and Negative Syndrome Scale, the Montgomery-Asberg Depression Rating Scale, the Arbeitsgemeinschaft Methodik und Dokumentation in der Psychiatrie scale for dissociation, and a Structured Clinical Interview for the Diagnostic and Statistical Manual of Mental Disorders, Revised Third Edition-based questionnaire. Multiple analysis of covariance was used to compare psychopathologic measures between groups with or without PTSD criteria A1 and A2, PTSD, and high and low levels of dissociation, controlling for substance abuse. RESULT: Trauma and dissociation were associated with more severe symptoms of schizophrenia. Especially high dissociation was associated with an increase in symptom load, whereas criterion A and PTSD had little or no such effect. DISCUSSION: In the present study, those with more severe schizophrenic and depressive symptoms were also characterized by high dissociation. The possible relation to trauma and schizophrenic diathesis is discussed along with the limitations of the study. PMID- 19216890 TI - Treatment of social anxiety with paroxetine: mediation of changes in anxiety and depression symptoms. AB - Investigation of relationship patterns between co-occurring symptoms has greatly improved the efficacy of psychiatric care. Depression and anxiety often present together, and identification of primary vs secondary psychiatric symptoms has implications for treatment. Previous psychotherapy research investigating the relationship between social anxiety and depression, across social anxiety treatment, found that severity of social anxiety accounted for most of the change in depression severity across time. Conversely, severity of depression accounted for little variation in severity of social anxiety. The current investigation was conducted to extend these findings by examining this mediational relationship in a pharmacologic trial comparing paroxetine (n = 20) and placebo (n = 22). Social anxiety and depression severity were assessed weekly for 16 weeks. Consistent with the previous study, results indicated that social anxiety severity mediated most of the variance in depression severity, with little variance accounted for by a test of the reverse mediation. Surprisingly, this same pattern was also found in the placebo group. These findings suggest that this pattern of mediational relationships may be fundamental to social anxiety, rather than specific to treatment modality or secondary comorbidity. PMID- 19216891 TI - Relationships between schizotypy and subjective experiences in a sample of 399 university students. AB - The high-risk approach based on the definition of schizotypy by Meehl (The American Psychologist 1962;17:827-38) and the basic symptom approach proposed by Huber (Fortschritte fur Neurologie und Psychiatrie 1957;25:491-520) share many points in common. In particular, several studies have shown that high scores on psychosis proneness or subjective experiences (SE) scales were associated with the onset of psychotic disorders in nonpsychiatric or nonpsychotic subjects. However, the relationships between the 2 concepts have never been studied experimentally. The present study was therefore designed to investigate the relationships between the scores of the SE scale and several psychosis proneness scales evaluating the negative and positive components of schizotypy in a sample of 399 university students. Data were analyzed using multiple regression and principal components analyses. The results show that SE, as assessed by the Frankfurt Complaint Questionnaire, were significantly associated with the positive component of schizotypy but not with the negative component. Subjective experiences could constitute vulnerability traits for psychosis and the Frankfurt Complaint Questionnaire could be use instead of the positive psychosis proneness scales. PMID- 19216892 TI - The impact of anxiety and migraine on quality of sleep in patients with major depressive disorder. AB - OBJECTIVE: The aim of this study was to investigate the impact of anxiety disorders and migraine on sleep quality and to find the independent factors that predict sleep quality among patients with major depressive disorder (MDD). METHOD: Psychiatric outpatients diagnosed with MDD were enrolled in the study. Major depressive disorder and 7 anxiety disorders were diagnosed using the Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision. Migraine was diagnosed based on the International Classification of Headache Disorders, Second Edition. Headache intensity and frequency were reported by the subjects. The Pittsburgh Sleep Quality Index and Hamilton Depression Rating Scale were used to evaluate quality of sleep and depression severity, respectively. Multiple linear regressions were used to identify independent factors related to sleep quality. RESULTS: One hundred thirty-five subjects (34 men and 101 women) with MDD were enrolled in the study. Subjects with panic disorder and agoraphobia were found to have poorer Pittsburgh Sleep Quality Index scores. Subjects with panic disorder, agoraphobia, and migraine had higher scores for items relating to sleep quality in the Hamilton Depression Rating Scale. Headache intensity and frequency correlated with sleep disturbance. Panic disorder was independently predictive of poor sleep quality. Both migraine and panic disorder independently predicted a greater severity of depression. CONCLUSION: Our study demonstrates the negative impact of panic disorder and migraine on MDD and some of the interrelations between depression, anxiety, and sleep quality. Future studies should further explore these interactions and consider possible therapeutic interventions. PMID- 19216893 TI - Are personality dimensions associated with sleep length in a large nationally representative sample? AB - OBJECTIVES: The purpose of this hypothesis-generating study was to determine whether personality domains and specific personality traits are uniquely associated with sleep duration using data obtained from the National Comorbidity Survey (NCS). METHODS: Using trained interviewers, we administered to the 5877 noninstitutionalized adults living in the United States the Composite International Diagnostic Interview (World Health Organization. Composite International Diagnostic Interview. Geneva, Switzerland: WHO, 1990) to assess for any DSM-III-R psychiatric diagnoses, and they completed self-report measures of personality and sleep. This was a secondary data analysis using information from a large existing public use data set (NCS-part II). The NCS-part II was an epidemiologic survey based on a stratified multistage area probability method with a response rate of 82.4%. RESULTS: Using a multivariate logistic regression technique, we found significant and positive associations between short sleep (defined as or=9 hours per 24 hour period) and a diagnosis of dysthymia (OR = 1.52), the use of a sedating medication in the past 12 months (OR = 1.52), emotional reliance on another person (OR = 1.37), employment status (OR = 1.31), and marital status (OR = 1.20). CONCLUSIONS: Findings suggest that personality, even after controlling for psychiatric and medical conditions, is associated with sleep length and may be an additional factor to consider when assessing any individual patient. PMID- 19216894 TI - Personality subtypes in patients with panic disorder. AB - OBJECTIVE: This study examined personality subtypes of adults diagnosed with panic disorder (PD). METHODS: As part of a National Institute of Mental Health funded study on personality in a clinical population, randomly selected psychologists and psychiatrists used a Q-sort instrument to describe 96 adults diagnosed with PD. RESULTS: Q-factor analysis yielded 4 personality subtypes: high functioning, emotionally dysregulated, inhibited/avoidant, and somatizing. Within the sample, the subtypes differed on Axis I and II pathology, adaptive functioning, and developmental and family history variables. Personality constellations in the sample showed substantial incremental validity in predicting adaptive functioning and treatment response above and beyond the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Axis I diagnoses. CONCLUSIONS: These distinctions may elucidate the high rates of comorbidity among patients with PD and are important in understanding treatment choice and outcome. PMID- 19216895 TI - Sex differences among treatment-seeking adult pathologic gamblers. AB - OBJECTIVE: The purpose of this study is to explore the effect of sex as a moderator variable for gambling and clinical profiles in a large sample of Spanish treatment-seeking patients for pathologic gambling (PG). METHOD: Clinical and personality profiles were compared between 143 male and 143 female pathologic gamblers who sought consultation at a specialized hospital unit. Multiple regressions explored the incremental predictive accuracy of sex on PG severity in consideration of sociodemographic and psychologic characteristics. RESULTS: Men gambled most frequently using slot machines and lotteries, spent more money, and had most arguments with family and friends. Although the age of onset of PG was 7.1 years higher for females, the severity was equal for both sexes. Women evidenced more general psychopathology, with higher mean scores in all the Symptom ChekList-90 items scales (except for hostility and psychoticism), and had significantly higher scores for harm avoidance and lower scores for self directedness than the male group. However, sex alone did not obtain a significant incremental validity for PG severity. CONCLUSIONS: These results may provide guidance for obtaining accurate diagnostic information about PG, properly identifying patients with specific needs and planning sex-specific targets. PMID- 19216896 TI - The 4-Point ordinal Alliance Self-report: a self-report questionnaire for assessing therapeutic relationships in routine mental health. AB - BACKGROUND: The quality of the relationship between patient and therapist can be considered a cornerstone of psychiatric practice. Nonadherence is one of the leading problems affecting antipsychotic effectiveness in schizophrenic patients and represents a poor prognostic factor for schizophrenia. Among the factors influencing adherence, the clinician's style of communication and the therapeutic relationship (TR) are recognized as key points. The measures of TR have been broadly explored in psychotherapy process but have received little attention in the treatment of severe mental illness in the context of routine mental health. Our objective was to build a self-rating scale easy to use in clinical psychiatric practice to assess TR, including drug-taking aspects and the relationship with the clinician on a day-to-day basis. A secondary objective was to assess the scale's validity and the association between TR and adherence. METHODS: The study was conducted with 92 consenting inpatients who met specified criteria for schizophrenia and schizoaffective disorders (Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition). Assessments of TR were obtained by the 4-Point ordinal Alliance Self-report and an 8-cm visual analog scale completed by the patient 1 week before discharge, after the remission of acute symptomatology. Adherence behavior was estimated with a new valid and reliable questionnaire called the Medication Adherence Rating Scale. Factor analysis was performed. RESULTS: Internal consistency was assessed by calculating Cronbach alpha, which revealed a highly satisfactory value (alpha = .91). After oblique rotation was run, 2 understandable factors were extracted that we termed empathy experienced and psychoeducation. CONCLUSIONS: Therapeutic relationship with the clinician can be considered a prerequisite for positive treatment course and outcome. The detection of subpopulations characterized by determinants of poor TR could be the first step toward improving schizophrenia prognosis linked to poor adherence. PMID- 19216897 TI - Differences in audiovisual integration, as measured by McGurk phenomenon, among adult and adolescent patients with schizophrenia and age-matched healthy control groups. AB - BACKGROUND: McGurk effect is a perceptual phenomenon that reflects the integration of visual and auditory information during speech perception. Using McGurk effect, the authors examined the audiovisual integration in adolescents and adults with schizophrenia as compared with healthy volunteers. SAMPLING AND METHODS: Thirty hospitalized patients with schizophrenia and 20 age-matched healthy controls were examined for perception of ambiguous audiovisual stimuli. RESULTS: The mean of McGurk-positive responses was significantly lower in adolescent patients with schizophrenia than in healthy adolescents (3.13 +/- 2.09 vs 5.60 +/- 0.7, respectively; t = 3.591, P = .001). The McGurk-positive responses were significantly higher in healthy adolescents than in healthy adults (5.60 +/- 0.7 vs 3.60 +/- 1.43, respectively; t = 3.974, P = .001). No significant difference in McGurk-positive responses was found between adults with schizophrenia and healthy adult individuals, or between adolescent and adults with schizophrenia. Duration of schizophrenia, soft sign status, type of symptoms, and type of antipsychotic medication showed no influence on the audiovisual integration ability. CONCLUSIONS: (I) Age effect: It seems that the audiovisual integrative function increases from childhood to adolescence and decreases from adolescence to early adulthood. (II) Schizophrenia: Audiovisual integration is poor in adolescent and adult patients with schizophrenia. Thus, it seems that schizophrenia is associated with early and persistent impairment in the development of the audiovisual integration ability. (III) Reliance on visual cue stimuli: Although several previous investigations concluded that patients with schizophrenia rely less on visual cue stimuli than healthy controls, our data suggest that this is true only for specific types of visual cue stimuli. PMID- 19216899 TI - Initiation of autophagy by photodynamic therapy. AB - Photodynamic therapy (PDT) involves the irradiation of photosensitized cells with light. Depending on localization of the photosensitizing agent, the process can induce photodamage to the endoplasmic reticulum (ER), mitochondria, plasma membrane, and/or lysosomes. When ER or mitochondria are targeted, antiapoptotic proteins of the Bcl-2 family are especially sensitive to photodamage. Both apoptosis and autophagy can occur after PDT, autophagy being associated with enhanced survival at low levels of photodamage to some cells. Autophagy can become a cell-death pathway if apoptosis is inhibited or when cells attempt to recycle damaged constituents beyond their capacity for recovery. While techniques associated with characterization of autophagy are generally applicable, PDT introduces additional factors related to unknown sites of photodamage that may alter autophagic pathways. This chapter discusses issues that may arise in assessing autophagy after cellular photodamage. PMID- 19216900 TI - Autophagic cell death. AB - In this chapter we discuss methods to study autophagic cell death. A large body of evidence demonstrates that autophagy is a cell survival mechanism in response to starvation. The role of autophagy in cell death, however, has long been controversial. Recently, molecular approaches have provided direct evidence that autophagy contributes to cell death in certain contexts. We begin this chapter by outlining methods to quantify cell death, for example by assaying for cell viability. Next, we discuss methods to measure processes involved in cell death, such as caspase activation and autophagy. Finally, we discuss methods to genetically or chemically perturb autophagy to test whether autophagy is required for cell death. Together, these approaches provide a guide to investigate the relationship between autophagy and cell death. PMID- 19216901 TI - Autophagic neuron death. AB - Neurons of the central nervous system (CNS) tissue are terminally differentiated cells and have large volumes, unlike cells of peripheral tissues. Such neurons possess abundant lysosomes in which damaged and unneeded intracellular constituents are degraded. A cellular process to bring the unneeded constituents to lysosomes is referred to as macroautophagy (autophagy), which is essential for the maintenance of cellular metabolism under physiological conditions. In fact, mice deficient in Atg7 or Atg5 specifically in CNS tissue have ubiquitin aggregates in neurons and massive loss of cerebral and cerebellar cortical neurons, resulting in neurodegeneration and short life span. In addition, acceleration of autophagy induced by the loss of lysosomal proteinases such as cathepsin D or cathepsins B and L, or by hypoxic/ischemic (H/I) brain injury, causes neurodegeneration. Moreover, lysosomes with undigested materials due to loss of proteinases are enwrapped by double membranes to produce autophagosomes, resulting in the further accumulation of autolysosomes. H/I brain injury at birth that is an important cause of cerebral palsy, mental retardation, and epilepsy causes energy failure, oxidative stress, and unbalanced ion fluxes, leading to a high induction of autophagy in brain neurons. Since mice that are unable to execute autophagy (due to brain-specific deletion of Atg7 or Atg5) die as a result of massive loss of cerebral and cerebellar neurons with accumulation of ubiquitin aggregates, induction of neuronal autophagy after H/I injury is generally considered neuroprotective, as it maintains cellular homeostasis. However, our data showing that H/I injury-induced pyramidal neuron death in the neonatal hippocampus is largely prevented by Atg7 deficiency indicate the presence of autophagic neuron death. In this section, we introduce various methods for the detection of autophagic neuron death in addition to other death modes of CNS neurons. PMID- 19216902 TI - Assessing metabolic stress and autophagy status in epithelial tumors. AB - Autophagy is a survival mechanism activated in response to metabolic stress. In normal tissues autophagy plays a major role in energy homeostasis through catabolic self-digestion of damaged proteins and organelles. Contrary to its survival function, autophagy defects are implicated in tumorigenesis suggesting that autophagy is a tumor suppression mechanism. Although the exact mechanism of this tumor suppressor function is not known, it likely involves mitigation of cellular damage leading to chromosomal instability. The complex role of functional autophagy in tumors calls for model systems that allow the assessment of autophagy status, stress management and the impact on oncogenesis both in vitro as well as in vivo. We developed model systems that involve generation of genetically defined, isogenic and immortal epithelial cells from different tissue types that are applicable to both wild-type and mutant mice. This permits the study of tissue- as well as gene-specific tumor promoting functions. We successfully employed this strategy to generate isogenic, immortal epithelial cell lines from wild-type and mutant mice deficient in essential autophagy genes such as beclin 1 (beclin 1(+/-)) and atg5 (atg 5(-/-)). As these cell lines are amenable to further genetic manipulation, they allowed us to generate cell lines with apoptosis defects and stable expression of the autophagy marker EGFP-LC3 that facilitate in vitro and in vivo assessment of stress-mediated autophagy induction. We applied this model system to directly monitor autophagy in cells and 3D-morphogenesis in vitro as well as in tumor allografts in vivo. Using this model system we demonstrated that autophagy is a survival response in solid tumors that co-localizes with hypoxic regions, allowing tolerance to metabolic stress. Furthermore, our studies have established that autophagy also protects tumor cells from genome damage and limits cell death and inflammation as possible means to tumor suppression. Additionally these cell lines provide an efficient way to perform biochemical analyses, and high throughput screening for modulators of autophagy for potential use in cancer therapy and prevention. PMID- 19216903 TI - Autophagic clearance of aggregate-prone proteins associated with neurodegeneration. AB - Autophagy has emerged as a field of rapidly growing interest with implications in several disease conditions, such as cancer, infectious diseases, and neurodegenerative diseases. Autophagy is a major degradation pathway for aggregate-prone, intracytosolic proteins causing neurodegenerative disorders, such as Huntington's disease and forms of Parkinson's disease. Up-regulating autophagy may be a tractable therapeutic intervention for clearing these disease causing proteins. The identification of autophagy-enhancing compounds would be beneficial not only in neurodegenerative diseases but also in other conditions where up-regulating autophagy may act as a protective pathway. Furthermore, small molecule modulators of autophagy may also be useful in dissecting pathways governing mammalian autophagy. In this chapter, we highlight assays that can be used for the identification of autophagy regulators, such as measuring the clearance of mutant aggregate-prone proteins or of autophagic flux with bafilomycin A(1). Using these methods, we recently described several mTOR independent autophagy-enhancing compounds that have protective effects in various models of Huntington's disease. PMID- 19216904 TI - Monitoring autophagy in Alzheimer's disease and related neurodegenerative diseases. AB - This chapter describes detailed methods to monitor autophagy in neurodegenerative disorders, especially in Alzheimer's disease. Strategies to assess the competence of autophagy-related mechanisms in disease states ideally incorporate analyses of human disease and control tissues, which may include brain, fibroblasts, or other peripheral cells, in addition to animal and cell models of the neurodegenerative disease pathology and pathobiology. Cross-validation of pathophysiological mechanisms in the diseased tissues is always critical. Because of the cellular heterogeneity of the brain and the differential vulnerability of the neural cells in a given disease state, analyses focus on regional comparisons of affected and unaffected regions or cell populations within a particular brain region and include ultrastructural, immunological, and cell and molecular biological approaches. PMID- 19216905 TI - Live-cell imaging of autophagy induction and autophagosome-lysosome fusion in primary cultured neurons. AB - The discovery that impaired autophagy is linked to a wide variety of prominent diseases including cancer and neurodegeneration has led to an explosion of research in this area. Methodologies that allow investigators to observe and quantify the autophagic process will clearly advance knowledge of how this process contributes to the pathophysiology of many clinical disorders. The recent identification of essential autophagy genes in higher eukaryotes has made it possible to analyze autophagy in mammalian cells that express autophagy proteins tagged with fluorescent markers. This chapter describes such methods using primary cultured neurons that undergo up-regulation of autophagy when trophic factors are removed from their medium. The prolonged up-regulated autophagy, in turn, contributes to the death of these neurons, thus providing a model to examine the relationship between enhanced autophagy and cell death. Neurons are isolated from the cerebellum of postnatal day 7 rat pups and cultured in the presence of trophic factors and depolarizing concentrations of potassium. Once established, the neurons are transfected with an adeno-viral vector expressing MAP1-LC3 with red fluorescent protein (RFP). MAP1-LC3 is the mammalian homolog of the yeast autophagosomal marker Atg8 and when tagged to GFP or RFP, it is the most widely used marker for autophagosomes. Once expression is stable, autophagy is induced by removing trophic factors. At various time points after inducing autophagy, the neurons are stained with LysoSensor Green (a pH-dependent lysosome marker) and Hoechst (a DNA marker) and subjected to live-cell imaging. In some cases, time-lapse imaging is used to examine the stepwise process of autophagy in live neurons. PMID- 19216906 TI - Using genetic mouse models to study the biology and pathology of autophagy in the central nervous system. AB - Autophagy is a cellular self-eating process that plays an important role in neuroprotection as well as neuronal injury and death. The detailed pathway of autophagy in these two opposing functions remains to be elucidated. Neurons are highly specialized, postmitotic cells that are typically composed of a soma (cell body), a dendritic tree, and an axon. Here, we describe methods for studying autophagy in the central nervous system (CNS). The first involves the use of recently developed transgenic mice expressing the fluorescent autophagosome marker, GFP-LC3. Although CNS neurons show little evidence for the presence of GFP-LC3-containing puncta under normal conditions, under pathological conditions such neurons exhibit many GFP-LC3 puncta. The onset and density of GFP-LC3 puncta have been found to vary significantly in the subcompartments of the affected neurons. These studies suggest that autophagy is distinctly regulated in CNS neurons and that neuronal autophagy can be highly compartmentalized. While transgenic mice expressing GFP-LC3 are a valuable tool for assessing autophagic activity in the CNS, caution needs to be taken when interpreting results solely based on the presence of GFP-LC3 puncta. Therefore, traditional ultrastructural analysis using electron microscopy remains an important tool for studying autophagosomes in vivo. Additional reporters of autophagy are constantly being sought. For example, recently a selective substrate of autophagy p62/SQSTM1 has been shown to be specifically regulated by autophagic activity. Therefore, p62/SQSTM1 protein levels can be used as an additional reporter for autophagic activity. PMID- 19216907 TI - Biochemical and morphological detection of inclusion bodies in autophagy deficient mice. AB - Autophagy-deficient mice exhibit the formation of ubiquitin-inclusions in the liver and brain, which is not attributed to the dysfunction of the ubiquitin proteasome system. Moreover, it is also clear that a multifunctional protein p62/A170/SQSTM1 (hereafter referred to as p62) links autophagy and inclusion formation, being one of the key components of the ubiquitin inclusions. The ubiquitin/p62 inclusions can be detected in the detergent-insoluble fraction by western blot analysis, while morphological information can be obtained by immunohistochemistry at both the light and electron microscopy levels. Importantly, p62 has become a reliable marker, with which we can identify inclusions and estimate autophagic activity in diseased tissues or cells. In this chapter, we describe the methods used for biochemical and morphological detection of ubiquitin/p62-inclusions in autophagy-suppressed Atg7-deficient mice. These methods are suitable for examination of cells and tissues with conditions associated with reduced autophagy (e.g., aging and mice models of intractable diseases such as Alzheimer's disease), and their applications should enhance our understanding of the pathophysiological mechanisms involved in the formation of intracellular inclusions. PMID- 19216908 TI - Analyzing autophagy in clinical tissues of lung and vascular diseases. AB - Autophagy, a process by which organelles and cellular proteins are encapsulated in double-membrane vesicles and subsequently degraded by lysosomes, plays a central role in cellular and tissue homeostasis. In various model systems, autophagy may be triggered by nutrient deprivation, oxidative stress, and other insults such as endoplasmic reticulum stress, hypoxia, and pathogen infection. The role of autophagy in lung physiology and homeostasis, however, has not been well studied. Even less is known of the role of autophagy in the pathogenesis of chronic lung disease. Autophagy may act essentially as a protective mechanism in lung cells, by removing dysfunctional organelles, and recycling essential nutrients. On the other hand, excessive autophagy may also contribute to cell death pathways, resulting in the depletion of critical cell populations, and thus may also contribute to the disease pathogenesis. An understanding of the cell type specific regulation and function of autophagy in the lung may facilitate the development of therapeutic strategies for the treatment of lung pathologies. This chapter provides protocols for the isolation of distinct lung cell types, such as epithelial, endothelial, macrophages, and fibroblasts; as well as protocols for the analysis of autophagy in lung cells and tissues. PMID- 19216909 TI - Autophagy in neurite injury and neurodegeneration: in vitro and in vivo models. AB - Recent advances indicate that maintaining a balanced level of autophagy is critically important for neuronal health and function. Pathologic dysregulation of macroautophagy has been implicated in synaptic dysfunction, cellular stress, and neuronal cell death. Autophagosomes and autolysosomes are induced in acute and chronic neurological disorders including stroke, brain trauma, neurotoxin injury, Parkinson's, Alzheimer's, Huntington's, motor neuron, prion, lysosomal storage, and other neurodegenerative diseases. Compared to other cell types, neuronal autophagy research presents particular challenges that may be addressed through still evolving techniques. Neuronal function depends upon maintenance of axons and dendrites (collectively known as neurites) that extend for great distances from the cell body. Both autophagy and mitochondrial content have been implicated in regulation of neurite length and function in physiological (plasticity) and pathological remodeling. Here, we highlight several molecular cell biological and imaging methods to study autophagy and mitophagy in neuritic and somatic compartments of differentiated neuronal cell lines and primary neuron cultures, using protocols developed in toxic and genetic models of parkinsonian neurodegeneration. In addition, mature neurons can be studied using in vivo protocols for modeling ischemic and traumatic injuries. Future challenges include application of automated computer-assisted image analysis to the axodendritic tree of individual neurons and improving methods for measuring neuronal autophagic flux. PMID- 19216910 TI - Monitoring the autophagy pathway in cancer. AB - Autophagy is an ancient cell survival pathway that is induced by metabolic stress and that helps prevent bioenergetic failure. This pathway has emerged as a promising new target in cancer treatment, where agents that inhibit autophagic degradation have efficacy in preventing cancer and in treating resistant disease when combined with conventional chemotherapeutics, which generally activate the pathway. However, agents that specifically target the autophagy pathway are currently lacking, and monitoring the effects of therapeutics on the autophagy pathway raises several challenges. Here we review the potential roles of the autophagy pathway in tumor progression and in maintenance of the malignant state, and introduce novel methods that we have developed that allow one to monitor autophagic activity ex vivo and in vivo. PMID- 19216911 TI - Autophagy pathways in glioblastoma. AB - Glioma cells are more likely to respond to therapy through autophagy than through apoptosis. The most efficacious cytotoxic drugs employed in glioma therapy, such as temozolomide and rapamycin, induce autophagy. Oncolytic adenoviruses, which will soon be tested in patients with gliomas at the University of Texas M. D. Anderson Cancer Center, also induce autophagy. Autophagy in gliomas thus represents a promising mechanism that may lead to new glioma therapies. In this chapter, we present the methods for studying autophagy in glioma cells, including assessment of in vitro cellular markers acidic vesicle organelles, and green fluorescent protein (GFP)-LC3 punctation; biochemical markers LC3-I/II conversion, p62 degradation, Atg12-Atg5 accumulation, and p70S6K dephosphorylation; and ultrastucture of the autophagosomes. In addition, we will address how LC3B and Atg5 up-regulation during autophagy can be examined through immunostaining in treated tumors and the potential of these proteins for use as surrogate markers to monitor therapeutic effects in clinical trials. Finally, we will discuss the challenges of studying autophagy in gliomas and the future directions of such use. PMID- 19216912 TI - Autophagy in lung cancer. AB - Lung cancer is the leading cause of cancer-related deaths worldwide. The relatively poor cure rate in lung cancer patients has been associated with a resistance to chemotherapy and radiation that is at least in part related to defects in cellular apoptotic machinery. Exploitation of another form of cell death, autophagy, has the capacity to improve the therapeutic gain of current therapies. In an effort to develop novel treatment strategies to enhance the therapeutic ratio for lung cancer, we wish to better understand the role of autophagic cell death for the sensitization of lung cancer. This text reviews the most up to date protocols and techniques for the study of autophagic cell death in lung cancer models. Others may use these techniques as a framework for study within their experimental models. PMID- 19216913 TI - Signal-dependent control of autophagy-related gene expression. AB - Several tumors arise from deregulated signaling pathways leading to increased proliferation and impairment of differentiation. To bypass endogenous control mechanisms and to survive the environmental stress associated with increased growth, tumor cells acquire a plethora of modifications that ultimately tend to down-regulate the ability to undergo apoptosis and exacerbate prosurvival mechanisms. Autophagy is an evolutionarily conserved mechanism through which cells recycle essential molecular constituents or eliminate damaged organelles under stress conditions imposed by nutrients or growth factors deprivation. As such, autophagy acts as a prosurvival mechanism for cancer cells. However, when overactivated, autophagy could also represent a cell death mechanism acting through self-cannibalization. Therefore, understanding the various signaling pathways that regulate autophagy could be of extreme importance. Indeed, the identification of specific molecular targets amenable to pharmacological manipulation to induce cancer cell self-cannibalization could represent a promising approach to treat apoptosis-resistant tumors. PMID- 19216914 TI - Novel methods for measuring cardiac autophagy in vivo. AB - Autophagy, a highly conserved cellular mechanism wherein various cellular components are broken down and recycled through lysosomes, occurs constitutively in the heart and may serve as a cardioprotective mechanism in some situations. It has been implicated in the development of heart failure and is up-regulated following ischemia-reperfusion injury. Autophagic flux, a measure of autophagic vesicle formation and clearance, is an important measurement in evaluating the efficacy of the pathway, however, tools to measure flux in vivo have been limited. Here, we describe the use of monodansylcadaverine (MDC) and the lysosomotropic drug chloroquine to measure autophagic flux in in vivo model systems, specifically focusing on its use in the myocardium. This method allows determination of flux as a more precise measure of autophagic activity in vivo much in the same way that Bafilomycin A(1) is used to measure flux in cell culture. MDC injected 1 h before sacrifice, colocalizes with mCherry-LC3 puncta, validating its use as a marker of autophagosomes. This chapter provides a method to measure autophagic flux in vivo in both transgenic and nontransgenic animals, using MDC and chloroquine, and in addition describes the mCherry-LC3 mouse and the advantages of this animal model in the study of cardiac autophagy. Additionally, we review several methods for inducing autophagy in the myocardium under pathological conditions such as myocardial infarction, ischemia/ reperfusion, pressure overloading, and nutrient starvation. PMID- 19216916 TI - Evaluation of cell death markers in severe calcified aortic valves. AB - Degenerative aortic valve disease is the most frequent acquired valve disease. Especially in the elderly, its prevalence is increasing. Once the disease becomes symptomatic, it is rapidly fatal. The disease cannot be considered a result of aging alone. The condition is an active process, which occurs with rapid progression, especially when calcification can be documented. This calcification can be the end result of cellular mechanisms involving cell death pathways (such as autophagy) and cellular matrix remodeling. These processes are beginning to be unraveled in the initiation and propagation of the disease. Autophagy could be the common step through which these mechanisms lead to this pathway of cell death in this disease. Autophagy can be detected by procedures described hereafter. PMID- 19216917 TI - Monitoring autophagy in muscle diseases. AB - Autophagy is a tightly regulated pathway for the degradation and recycling of proteins delivered to lysosomes, and is an important process in maintaining cellular homeostasis. Whereas a basal level of autophagy can be detected in skeletal muscles, its perturbation can be seen in a variety of conditions affecting the muscle. In certain muscle diseases, moreover, autophagy seems to be a characteristic feature, although the exact role of autophagy in these disorders is just starting to be understood. As autophagy is not only an index of disease progression but also a potential target for treatment in certain disease conditions, its characterization is indeed of relevance. Thus, in this chapter, methods applicable to both human and murine skeletal muscle preparation for the analysis and monitoring of autophagy are presented. PMID- 19216915 TI - Autophagy in load-induced heart disease. AB - The heart is a highly plastic organ capable of remodeling in response to changes in physiological or pathological demand. When workload increases, the heart compensates through hypertrophic growth of individual cardiomyocytes to increase cardiac output. However, sustained stress, such as occurs with hypertension or following myocardial infarction, triggers changes in sarcomeric protein composition and energy metabolism, loss of cardiomyocytes, ventricular dilation, reduced pump function, and ultimately heart failure. It has been known for some time that autophagy is active in cardiomyocytes, occurring at increased levels in disease. Yet the potential contribution of cardiomyocyte autophagy to ventricular remodeling and disease pathogenesis has only recently been explored. This latter fact stems largely from the recent emergence of tools to probe molecular mechanisms governing cardiac plasticity and to define the role of autophagic flux in the context of heart disease. In this chapter, we briefly review prominent mouse models useful in the study of load-induced heart disease and standard techniques used to assess whether a molecular or cellular event is adaptive or maladaptive. We then outline methods available for monitoring autophagic activity in the heart, providing detailed protocols for several techniques unique to working with heart and other striated muscles. PMID- 19216918 TI - Analyzing macroautophagy in hepatocytes and the liver. AB - Mammalian autophagy has been well characterized in the liver and in hepatocytes. Autophagy plays important roles in the normal physiology of the liver and in the pathogenesis of several liver diseases. This chapter will discuss the commonly used methods for analysis of autophagy in hepatocytes and in the liver. PMID- 19216920 TI - Computer Methods, Part A. Preface. PMID- 19216919 TI - Monitoring autophagy in lysosomal storage disorders. AB - Lysosomes are the final destination of the autophagic pathway. It is in the acidic milieu of the lysosomes that autophagic cargo is metabolized and recycled. One would expect that diseases with primary lysosomal defects would be among the first systems in which autophagy would be studied. In reality, this is not the case. Lysosomal storage diseases, a group of more than 60 diverse inherited disorders, have only recently become a focus of autophagic research. Studies of these clinically severe conditions promise not only to clarify pathogenic mechanisms, but also to expand our knowledge of autophagy itself. In this chapter, we will describe the lysosomal storage diseases in which autophagy has been explored, and present the approaches used to evaluate this essential cellular pathway. PMID- 19216921 TI - Phase response curves elucidating the dynamics of coupled oscillators. AB - Phase response curves (PRCs) are widely used in circadian clocks, neuroscience, and heart physiology. They quantify the response of an oscillator to pulse-like perturbations. Phase response curves provide valuable information on the properties of oscillators and their synchronization. This chapter discusses biological self-sustained oscillators (circadian clock, physiological rhythms, etc.) in the context of nonlinear dynamics theory. Coupled oscillators can synchronize with different frequency ratios, can generate toroidal dynamics (superposition of independent frequencies), and may lead to deterministic chaos. These nonlinear phenomena can be analyzed with the aid of a phase transition curve, which is intimately related to the phase response curve. For illustration purposes, this chapter discusses a model of circadian oscillations based on a delayed negative feedback. In a second part, the chapter provides a step-by-step recipe to measure phase response curves. It discusses specifications of this recipe for circadian rhythms, heart rhythms, neuronal spikes, central pattern generators, and insect communication. Finally, it stresses the predictive power of measured phase response curves. PRCs can be used to quantify the coupling strength of oscillations, to classify oscillator types, and to predict the complex dynamics of periodically driven oscillations. PMID- 19216923 TI - Analytical methods for the retrieval and interpretation of continuous glucose monitoring data in diabetes. AB - Scientific and industrial effort is now increasingly focused on the development of closed-loop control systems (artificial pancreas) to control glucose metabolism of people with diabetes, particularly type 1 diabetes mellitus. The primary prerequisite to a successful artificial pancreas, and to optimal diabetes control in general, is the continuous glucose monitor (CGM), which measures glucose levels frequently (e.g., every 5 min). Thus, a CGM collects detailed glucose time series, which carry significant information about the dynamics of glucose fluctuations. However, a CGM assesses blood glucose indirectly via subcutaneous determinations. As a result, two types of analytical problems arise for the retrieval and interpretation of CGM data: (1) the order and the timing of CGM readings and (2) sensor errors, time lag, and deviations from BG need to be accounted for. In order to improve the quality of information extracted from CGM data, we suggest several analytical and data visualization methods. These analyses evaluate CGM errors, assess risks associated with glucose variability, quantify glucose system stability, and predict glucose fluctuation. All analyses are illustrated with data collected using MiniMed CGMS (Medtronic, Northridge, CA) and Freestyle Navigator (Abbott Diabetes Care, Alameda, CA). It is important to remember that traditional statistics do not work well with CGM data because consecutive CGM readings are highly interdependent. In conclusion, advanced analysis and visualization of CGM data allow for evaluation of dynamical characteristics of diabetes and reveal clinical information that is inaccessible via standard statistics, which do not take into account the temporal structure of data. The use of such methods has the potential to enable optimal glycemic control in diabetes and, in the future, artificial pancreas systems. PMID- 19216922 TI - Multiple ion binding equilibria, reaction kinetics, and thermodynamics in dynamic models of biochemical pathways. AB - The operation of biochemical systems in vivo and in vitro is strongly influenced by complex interactions between biochemical reactants and ions such as H(+), Mg(2+), K(+), and Ca(2+). These are important second messengers in metabolic and signaling pathways that directly influence the kinetics and thermodynamics of biochemical systems. Herein we describe the biophysical theory and computational methods to account for multiple ion binding to biochemical reactants and demonstrate the crucial effects of ion binding on biochemical reaction kinetics and thermodynamics. In simulations of realistic systems, the concentrations of these ions change with time due to dynamic buffering and competitive binding. In turn, the effective thermodynamic properties vary as functions of cation concentrations and important environmental variables such as temperature and overall ionic strength. Physically realistic simulations of biochemical systems require incorporating all of these phenomena into a coherent mathematical description. Several applications to physiological systems are demonstrated based on this coherent simulation framework. PMID- 19216924 TI - Analysis of heterogeneity in molecular weight and shape by analytical ultracentrifugation using parallel distributed computing. AB - A computational approach for fitting sedimentation velocity experiments from an analytical ultracentrifuge in a model-independent fashion is presented. This chapter offers a recipe for obtaining high-resolution information for both the shape and the molecular weight distributions of complex mixtures that are heterogeneous in shape and molecular weight and provides suggestions for experimental design to optimize information content. A combination of three methods is used to find the solution most parsimonious in parameters and to verify the statistical confidence intervals of the determined parameters. A supercomputer implementation with a MySQL database back end is integrated into the UltraScan analysis software. The UltraScan LIMS Web portal is used to perform the calculations through a Web interface. The performance and limitations of the method when employed for the analysis of complex mixtures are demonstrated using both simulated data and experimental data characterizing amyloid aggregation. PMID- 19216926 TI - Analyses for physiological and behavioral rhythmicity. AB - Biological data that contain cycles require specialized statistical and analytical procedures. Techniques for analysis of time series from three types of systems are considered with the intent that the choice of examples is sufficiently broad that the processes described can be generalized to most other types of physiological or behavioral work. Behavioral circadian rhythms, acoustic signals in fly mating, and the Drosophila melanogaster cardiac system have been picked as typical in three broad areas. Worked examples from the fly cardiac system are studied in full detail throughout. The nature of the data streams and how they are acquired is first discussed with attention paid to ensuring satisfactory subsequent statistical treatment. Analysis in the time domain, namely simple and advanced plotting of data, autocorrelation analysis, and cross correlation, is described. The search for periodicity is conducted through examples of analysis in the frequency domain, primarily spectral analysis. Nonstationary time series pose a particular problem, and wavelet analysis of Drosophila mating song is described in detail as an example. Conditioning of data to improve output with digital filters, Fourier filtering, and trend removal is described. Finally, two tests for noise levels and regularity are considered. All the nonproprietary software used throughout the work is available from the author free of charge and can be specifically tailored to the needs of individual systems. PMID- 19216925 TI - Discrete stochastic simulation methods for chemically reacting systems. AB - Discrete stochastic chemical kinetics describe the time evolution of a chemically reacting system by taking into account the fact that, in reality, chemical species are present with integer populations and exhibit some degree of randomness in their dynamical behavior. In recent years, with the development of new techniques to study biochemistry dynamics in a single cell, there are increasing studies using this approach to chemical kinetics in cellular systems, where the small copy number of some reactant species in the cell may lead to deviations from the predictions of the deterministic differential equations of classical chemical kinetics. This chapter reviews the fundamental theory related to stochastic chemical kinetics and several simulation methods based on that theory. We focus on nonstiff biochemical systems and the two most important discrete stochastic simulation methods: Gillespie's stochastic simulation algorithm (SSA) and the tau-leaping method. Different implementation strategies of these two methods are discussed. Then we recommend a relatively simple and efficient strategy that combines the strengths of the two methods: the hybrid SSA/tau-leaping method. The implementation details of the hybrid strategy are given here and a related software package is introduced. Finally, the hybrid method is applied to simple biochemical systems as a demonstration of its application. PMID- 19216927 TI - A computational approach for the rational design of stable proteins and enzymes: optimization of surface charge-charge interactions. AB - The design of stable proteins and enzymes is not only of particular biotechnological importance, but also addresses some important fundamental questions. While there are a number of different options available for designing or engineering stable proteins, the field of computational design provides fast and universal methods for stabilizing proteins of interest. One of the successful computational design strategies focuses on stabilizing proteins through the optimization of charge-charge interactions on the protein surface. By optimizing surface interactions, it is possible to alleviate some of the challenges that accompany efforts to redesign the protein core. The rational design of surface charge-charge interactions also allows one to optimize only the interactions that are distant from binding sites or active sites, making it possible to increase stability without adversely affecting activity. The optimization of surface charge-charge interactions is discussed in detail along with the experimental evidence to demonstrate that this is a robust and universal approach to designing proteins with enhanced stability. PMID- 19216928 TI - Efficient computation of confidence intervals for Bayesian model predictions based on multidimensional parameter space. AB - A new algorithm is introduced to efficiently estimate confidence intervals for Bayesian model predictions based on multidimensional parameter space. The algorithm locates the boundary of the smallest confidence region in the multidimensional probability density function (pdf) for the model predictions by approximating a one-dimensional slice through the mode of the pdf with splines made of pieces of normal curve with continuous z values. This computationally efficient process (of order N) reduces estimation of the lower and upper bounds of the confidence interval to a multidimensional constrained nonlinear optimization problem, which can be solved with standard numerical procedures (of order N(2) or less). Application of the new algorithm is illustrated with a five dimensional example involving the computation of 95% confidence intervals for predictions made with a Bayesian forecasting model for cognitive performance deficits of sleep-deprived individuals. PMID- 19216929 TI - Analyzing enzymatic pH activity profiles and protein titration curves using structure-based pKa calculations and titration curve fitting. AB - The pH dependence of protein biophysical characteristics is often analyzed to gain an improved understanding of protein stability, enzyme activity, and protein ligand-binding processes. Indeed, much of our understanding of the catalytic mechanisms of enzymes derives from studies of the pH dependence of catalytic activity, and the ability to redesign the pH-dependent properties of enzymes continues to be of high relevance for both industrial and medical applications of proteins. This chapter discusses current theoretical methods for calculating protein pK(a) values and illustrates how one can analyze protein pK(a) calculation results to study calculation accuracy, pH stability profiles, and enzymatic pH activity profiles. A description of how one can analyze the importance of individual titratable groups is presented along with details on methods for redesigning protein pK(a) values and enzymatic pH activity profiles. Finally, I discuss novel methods for fitting experimental nuclear magnetic resonance titration curves and enzymatic pH activity profiles that can be used to derive information on electrostatic interaction energies in proteins. PMID- 19216930 TI - Least squares in calibration: weights, nonlinearity, and other nuisances. AB - In the least-squares fitting of data, there is a unique answer to the question of how the data should be weighted: inversely as their variance. Any other weighting gives less than optimal efficiency and leads to unreliable estimates of the parameter uncertainties. In calibration, knowledge of the data variance permits exact prediction of the precision of calibration, empowering the analyst to critically examine different response functions and different data structures. These points are illustrated here with a nonlinear response function that exhibits a type of saturation curvature at large signal like that observed in a number of detection methods. Exact error propagation is used to compute the uncertainty in the fitted response function and to treat common data transformations designed to reduce or eliminate the effects of data heteroscedasticity. Data variance functions can be estimated with adequate reliability from remarkably small data sets, illustrated here with three replicates at each of seven calibration values. As a quantitative goodness-of-fit indicator, chi(2) is better than the widely used R(2); in one application it shows clearly that the dominant source of uncertainty is not the measurement but the preparation of the calibration samples, forcing the conclusion that the calibration regression should be reversed. PMID- 19216931 TI - Evaluation and comparison of computational models. AB - Computational models are powerful tools that can enhance the understanding of scientific phenomena. The enterprise of modeling is most productive when the reasons underlying the adequacy of a model, and possibly its superiority to other models, are understood. This chapter begins with an overview of the main criteria that must be considered in model evaluation and selection, in particular explaining why generalizability is the preferred criterion for model selection. This is followed by a review of measures of generalizability. The final section demonstrates the use of five versatile and easy-to-use selection methods for choosing between two mathematical models of protein folding. PMID- 19216932 TI - Desegregating undergraduate mathematics and biology--interdisciplinary instruction with emphasis on ongoing biomedical research. AB - The remarkable advances in the field of biology in the last decade, specifically in the areas of biochemistry, genetics, genomics, proteomics, and systems biology, have demonstrated how critically important mathematical models and methods are in addressing questions of vital importance for these disciplines. There is little doubt that the need for utilizing and developing mathematical methods for biology research will only grow in the future. The rapidly increasing demand for scientists with appropriate interdisciplinary skills and knowledge, however, is not being reflected in the way undergraduate mathematics and biology courses are structured and taught in most colleges and universities nationwide. While a number of institutions have stepped forward and addressed this need by creating and offering interdisciplinary courses at the juncture of mathematics and biology, there are still many others at which there is little, if any, interdisciplinary interaction between the curricula. This chapter describes an interdisciplinary course and a textbook in mathematical biology developed collaboratively by faculty from Sweet Briar College and the University of Virginia School of Medicine. The course and textbook are designed to provide a bridge between the mathematical and biological sciences at the lower undergraduate level. The course is developed for and is being taught in a liberal arts setting at Sweet Briar College, Virginia, but some of the advanced modules are used in a course at the University of Virginia for advanced undergraduate and beginning graduate students. The individual modules are relatively independent and can be used as stand-alone projects in conventional mathematics and biology courses. Except for the introductory material, the course and textbook topics are based on current biomedical research. PMID- 19216933 TI - Mathematical algorithms for high-resolution DNA melting analysis. AB - This chapter discusses mathematical and computational methods that enhance the modeling, optimization, and analysis of high-resolution DNA melting assays. In conjunction with recent improvements in reagents and hardware, these algorithms have enabled new closed-tube techniques for genotyping, mutation scanning, confirming or ruling out genotypic identity among living related organ donors, and quantifying constituents in samples containing different DNA sequences. These methods are rapid, involving only 1 to 10 min of automatic fluorescence acquisition after a polymerase chain reaction. They are economical because inexpensive fluorescent dyes are used rather than fluorescently labeled probes. They are contamination-free and nondestructive. Specific topics include methods for extracting accurate melting curve information from raw signal, for clustering and classifying the results, for predicting complete melting curves and not just melting temperatures, and for modeling and analyzing the behavior of mixtures of multiple duplexes. PMID- 19216935 TI - AutoDecon: a robust numerical method for the quantification of pulsatile events. AB - This work presents a new approach to the analysis of aperiodic pulsatile heteroscedastic time-series data, specifically hormone pulsatility. We have utilized growth hormone (GH) concentration time-series data as an example for the utilization of this new algorithm. While many previously published approaches used for the analysis of GH pulsatility are both subjective and cumbersome to use, AutoDecon is a nonsubjective, standardized, and completely automated algorithm. We have employed computer simulations to evaluate the true-positive, the false-positive, the false-negative, and the sensitivity percentages of several of the routinely employed algorithms when applied to GH concentration time-series data. Based on these simulations, it was concluded that this new algorithm provides a substantial improvement over the previous methods. This novel method has many direct applications in addition to hormone pulsatility, for example, to time-domain fluorescence lifetime measurements, as the mathematical forms that describe these experimental systems are both convolution integrals. PMID- 19216934 TI - Biomathematical modeling of pulsatile hormone secretion: a historical perspective. AB - Shortly after the recognition of the profound physiological significance of the pulsatile nature of hormone secretion, computer-based modeling techniques were introduced for the identification and characterization of such pulses. Whereas these earlier approaches defined perturbations in hormone concentration-time series, deconvolution procedures were subsequently employed to separate such pulses into their secretion event and clearance components. Stochastic differential equation modeling was also used to define basal and pulsatile hormone secretion. To assess the regulation of individual components within a hormone network, a method that quantitated approximate entropy within hormone concentration-times series was described. To define relationships within coupled hormone systems, methods including cross-correlation and cross-approximate entropy were utilized. To address some of the inherent limitations of these methods, modeling techniques with which to appraise the strength of feedback signaling between and among hormone-secreting components of a network have been developed. Techniques such as dynamic modeling have been utilized to reconstruct dose-response interactions between hormones within coupled systems. A logical extension of these advances will require the development of mathematical methods with which to approximate endocrine networks exhibiting multiple feedback interactions and subsequently reconstruct their parameters based on experimental data for the purpose of testing regulatory hypotheses and estimating alterations in hormone release control mechanisms. PMID- 19216936 TI - Modeling fatigue over sleep deprivation, circadian rhythm, and caffeine with a minimal performance inhibitor model. AB - Sleep loss, as well as concomitant fatigue and risk, is ubiquitous in today's fast-paced society. A biomathematical model that succeeds in describing performance during extended wakefulness would have practical utility in operational environments and could help elucidate the physiological basis of sleep loss effects. Eighteen subjects (14 males, 4 females; age 25.8 +/- 4.3 years) with low levels of habitual caffeine consumption (<300 mg/day) participated. On night 1, subjects slept for 8 h (2300-0700 h), followed by 77 h of continuous wakefulness. They were assigned randomly to receive placebo or caffeine (200 mg, i.e., two sticks of Stay Alert gum) at 0100, 0300, 0500, and 0700 during nights 2, 3, and 4. The psychomotor vigilance test (PVT) was administered periodically over the 77-h period of continuous wakefulness. Statistical analysis reveals lognormality in each PVT, allowing for closed-form median calculation. An iterative parameter estimation algorithm, which takes advantage of MatLab's (R2007a) least-squares nonlinear regression, is used to estimate model parameters from subjects' PVT medians over time awake. In the model, daily periodicity is accounted for with a four-component Fourier series, and a simplified binding function describes asymptotic fatigue. The model highlights patterns in data that suggest (1) the presence of a performance inhibitor that increases and saturates over the period of continuous wakefulness, (2) competitive inhibition of this inhibitor by caffeine, (3) the persistence of an internally driven circadian rhythm of alertness, and (4) a multiplicative relationship between circadian rhythm and performance inhibition. The present inhibitor-based minimal model describes performance data in a manner consistent with known biochemical processes. PMID- 19216937 TI - Apyrase-based colorimetric test for detection of Shigella and enteroinvasive Escherichia coli in stool. AB - For lack of simple inexpensive early detection methods for Shigella spp. and enteroinvasive Escherichia coli (EIEC), bacillary dysentery remains a major cause of childhood mortality and morbidity in India and other developing countries. Rapid stool testing for apyrase, a specific periplasmic enzyme essential for the pathogen's intracellular spread, may provide a solution. We have developed a whole-cell colorimetric pyrophosphate hydrolysis assay based on cheap, stable, and locally available reagents. An innovative filtration-cum-inoculation step eliminates interfering stool solids and ensures sufficient bacterial growth and apyrase expression in 6 to 7 h at 37 degrees C. In a limited double-blind study of 57 clinical isolates of common enterobacteria, the test showed 100% sensitivity and 80% specificity for Shigella spp. and EIEC. Requiring only widely available equipment and inexpensive consumables, this affordable test is readily adaptable for determining antibiograms and for surveillance of food and water samples for the presence of Shigella and EIEC. PMID- 19216938 TI - Validation of an immunologic diagnostic kit for infectious vaginitis by Trichomonas vaginalis, Candida spp., and Gardnerella vaginalis. AB - FemPure is a kit for the rapid diagnosis of vaginitis by Trichomonas vaginalis, Candida spp., and Gardnerella vaginalis, based on aggregation of latex particles joined to specific antibodies. The validation of the method involved the parameters specificity, detection limit, robustness, clinical sensitivity, and clinical specificity. Also, samples analyzed in parallel by the validated test and other recognized tests conducted by external laboratory were included. The method was specific for the 3 infectious agents, and no cross-reaction with other microorganisms usually present in vaginal exudates. The detection limit > or =1 x 10(6) CFU/mL for Candida albicans and G. vaginalis avoids the detection of concentrations considered normal flora, whereas T. vaginalis was detected until 1 x 10(5) cells/mL. Values of clinical sensitivity > or =80% and clinical specificity > or =90% and concordance > or =90% were found between samples evaluated in parallel by different methods. Robustness showed that the test can be used in laboratories with different management systems; its simple implementation without equipment allows the use in primary health care areas. PMID- 19216939 TI - Molecular analysis of diversity within the genus Pseudomonas in the lungs of cystic fibrosis patients. AB - The genus Pseudomonas contains species that can act as human pathogens and are important in many infections such as those occurring in the lungs of many cystic fibrosis (CF) patients. Current methodologies that rely on the cultivation of bacteria are too cumbersome and often lack sufficient discriminatory power to define the diversity of Pseudomonas spp. As a result, molecular-based approaches have many advantages when attempting to differentiate between species of this genus. This study assessed the ability of terminal restriction fragment length polymorphism (T-RFLP) profiling of the 16S-23S rRNA ITS1 gene region to differentiate species of the genus Pseudomonas. Before application to clinical samples, this approach was validated on a panel of 10 different Pseudomonas spp. T-RFLP profiling of the 16S-23S rRNA ITS1 gene region amplified from these strains differentiated all Pseudomonas spp. tested. The presence of Pseudomonas spp. in CF sputum was assessed through the detection of this ITS1 gene region as amplified from DNA extracted from 40 samples of CF sputum. The ITS1 gene region was detected in 75% of these samples, including 5 from which no Pseudomonas spp. had been identified using culture-based methods. In silico analysis showed that all sequences amplified had a high homology with the ITS1 region of Pseudomonas aeruginosa. T-RFLP gave data that were consistent with the band being generated from P. aeruginosa for each patient. No correlation between Pseudomonas diversity and severity of lung disease was observed in this group of CF patients. This study has however shown that molecular analysis of the ITS1 region is effective in resolving diversity within the genus Pseudomonas. The wider use of this application is discussed. PMID- 19216940 TI - Molecular identification of Mycobacterium chimaera as a cause of infection in a patient with chronic obstructive pulmonary disease. AB - This report describes a case of Mycobacterium chimaera infection in a patient with a history of chronic obstructive pulmonary disease where the organism was identified by using molecular methods. M. chimaera was identified from fresh lung tissue and from an instrument-negative mycobacterial growth indicator tube broth culture. The utility of using sequence analysis of the internal transcribed spacer region for the rapid identification of a slow-growing nontuberculous Mycobacterium spp. where conventional culture methods were not successful was shown. PMID- 19216941 TI - Epidemiology and susceptibilities of methicillin-resistant Staphylococcus aureus in Taiwan: emphasis on chlorhexidine susceptibility. AB - Chlorhexidine is an antiseptic agent used for hand hygiene worldwide. To evaluate the susceptibility of methicillin-resistant Staphylococcus aureus (MRSA) to chlorhexidine, this study determined MICs of chlorhexidine and another 12 antimicrobial agents, carriage of the Panton-Valentine leukocidin, qacA/B, and smr genes, genetic relatedness by multilocus sequence typing (MLST), and staphylococcal cassette chromosome mec element type for 206 MRSA clinical isolates from the Taiwan Surveillance of Antimicrobial Resistance program III and IV (years 2002 and 2004) from 26 hospitals. Using MLST, we respectively identified 102 (49.5%), 68 (33.0%), 13 (6.3%), 5 (2.4%), 5 (2.4%), and 13 (6.3%) isolates as ST239, ST59, ST5, ST241, ST573, and other types. The MIC(50) and MIC(90) of chlorhexidine for all 206 isolates were 2 and 8 microg/mL, respectively. Seventy-three (35.4%) isolates carried qacA/B gene, but none carried smr. For the 72 (35.0%) MRSA isolates with chlorhexidine MIC > or =4 microg/mL, 53 were ST239 (49 of them carried qacA gene), 12 were ST5 (all carried qacB gene), 5 were ST241 (4 carried qacA gene), 1 was ST338 (and carried qacA gene), and 1 was ST573 (and carried qacA gene). Compared with other sequence-type MRSA isolates, ST239 MRSA isolates were the most resistant to both chlorhexidine and other antimicrobial agents. Methicillin-resistant S. aureus strains with disinfectant resistance qacA/B genes are common in Taiwan. High frequency of qacA/B genes among specific sequence types (ST239, ST5, and ST241) resulted in low susceptibility to chlorhexidine. Periodic surveillance of antiseptic susceptibility among MRSA isolates is important for the control of nosocomial hospital-acquired infections. PMID- 19216942 TI - Prevalence and characteristics of aac(6')-Ib-cr in AmpC-producing Enterobacter cloacae, Citrobacter freundii, and Serratia marcescens: a multicenter study from Korea. AB - We investigated the prevalence of aac(6')-Ib-cr and its association with other resistance genes in AmpC-producing Enterobacteriaceae without any selection criteria. A total of 479 clinical isolates of Enterobacter cloacae (179), Citrobacter freundii (134), and Serratia marcescens (166) from 12 laboratories between March and July 2005 were examined. We performed polymerase chain reaction for aac(6')-Ib, bla(OXA-1), ISEcp1, and class 1 integron. The aac(6')-Ib-cr was further identified by digestion with BstF5I and sequencing. The aac(6')-Ib was detected in 110 (23%) of 479 isolates, and 15 isolates (3.1%) were cr variants (8 E. cloacae, 5 C. freundii, and 2 S. marcescens). The aac(6')-Ib-cr was significantly associated with various resistance genes (bla(OXA-1), qnrS, qnrA, bla(CTX-M-3), and bla(CTX-M-14)), mobile elements (ISEcp1, ISCR1, and class 1 integron), and quinolone resistance. Eleven of 15 aac(6')-Ib-cr producers coharbored qnr genes. Although aac(6')-Ib-cr was uncommon in Korean AmpC producers, its association with various resistance genes and mobile elements would facilitate the dissemination of this variant. PMID- 19216943 TI - Mechanisms of resistance and mobility among multidrug-resistant CTX-M-producing Escherichia coli from Canadian intensive care units: the 1st report of QepA in North America. AB - We studied the molecular mechanisms of resistance and mobility of 18 multidrug resistant CTX-M-producing Escherichia coli isolates isolated from patients in Canadian intensive care units. Fluoroquinolone-resistant isolates (83.3%) had mutations in gyrA and parC. Plasmid-mediated quinolone resistance genes qnr (A, B, and S), qepA, and aac(6')-Ib-cr were detected in 0%, 5.6%, and 44.4%, respectively. Sulfamethoxazole/trimethoprim-resistant isolates (61.1%) carried a dfr gene, and 10 (90.9%) of the 11 carried 1 or more sul genes. Gentamicin resistant isolates (27.8%) carried the aac(3')-II gene, and doxycycline-resistant isolates (33.3%) carried 1 or more tet efflux genes. Both genetically related and unrelated groups of E. coli harboring extended-spectrum beta-lactamases were observed. The bla(CTX-M) genes were primarily located on diverse IncF plasmids of multiple replicon types downstream of the ISEcp1 element. The spread of the bla(CTX-M) genes among E. coli in Canada occurs through a diversity of different mechanisms and does not correspond to a single CTX-M determinant, or a single clone, or a single plasmid but rather through the combination of clonal spread of virulent strains and acquisition of diverse CTX-M-bearing plasmids. We report the 1st qepA-producing E. coli in North America. PMID- 19216944 TI - In vitro activity of iclaprim and comparison agents tested against Neisseria gonorrhoeae including medium growth supplement effects. AB - Iclaprim is a novel diaminopyrimidine currently in phase III clinical development. This study was conducted to determine the activity of iclaprim compared with trimethoprim and other commonly prescribed drug classes against a comprehensive collection of 156 Neisseria gonorrhoeae, including subsets of organisms resistant to beta-lactams and fluoroquinolones (>60%). Iclaprim (MIC(50/90), 4/8 microg/mL) was 16-fold more potent than trimethoprim (MIC(50/90), 64/>64 microg/mL), and medium growth supplements did not adversely influence activity. Lack of cross-resistances for iclaprim with other commonly used therapies could make iclaprim an alternative for several sexually transmitted diseases (gonococci, Chlamydia trachomatis). PMID- 19216945 TI - First report of acute purulent pericarditis by capsule genotype K1 Klebsiella pneumoniae in an alcoholic patient. PMID- 19216946 TI - Radiation burden of patients undergoing endovascular abdominal aortic aneurysm repair. AB - INTRODUCTION: Endovascular repair of abdominal aortic aneurysm (EVAR) requires the patient's extended exposure to x-rays, before, during, and after the intervention. The aim of this study was to determine the radiation exposure of patients undergoing EVAR and to assess the probability for the induction of both late and early radiation-related effects. METHODS: During the period of May 2006 to December 2007 EVAR was carried out in 62 patients using a mobile C-arm unit. The following dosimetric quantities were assessed: fluoroscopy time, cumulative dose in air, dose-area product, field area, and peak skin dose. RESULTS: The duration of fluoroscopy and the body mass index were found to be the main factors that influence the radiation burden in our hospital. The mean effective dose per procedure, 6.2 mSv, was between that from a planar coronary angiography and a coronary angioplasty. Taking into account the computed tomography (CT) procedure related angiographies carried out during the first year, patients receive a total effective dose of about 62 mSv within the first year. In vivo dosimetry showed that the peak skin dose was linearly correlated with cumulative dose in air and did not exceed 1.0 Gy, ie, it was less than the threshold for any acute skin reaction. CONCLUSION: Repair of abdominal aortic aneurysm results in substantial radiation burden. Radiation-related risks for carcinogenesis and skin injuries are factors that have to be taken into account in the selection of the strategy of each facility. PMID- 19216949 TI - Carotid artery stenting is safe and associated with comparable outcomes in men and women. AB - OBJECTIVE: Historically, large randomized controlled studies looking at carotid endarterectomy (CEA) have indicated an increased perioperative risk for women when gender subgroup analysis was performed. However, the outcomes of carotid stenting in women as compared to men have not been adequately investigated. We sought to compare the safety and efficacy of carotid angioplasty and stenting (CAS) when performed in women as compared to men. METHODS: Procedures, complications, demographics, co-morbidities, and follow-up data from carotid stenting procedures performed in a bi-campus division were entered into a prospective database and then retrospectively supplemented with stored angiographic image data and reviewed. Arterial anatomic characteristics evaluated using angiographic images were: common carotid/internal carotid lesion length ratio, common carotid/internal carotid diameter, index lesion length, common carotid/internal carotid artery tortuosity, and lesion and aortic arch calcification. Outcomes compared included groin complications, postoperative pressor requirements, length of stay, restenosis, stroke, myocardial infarction (MI), and death. RESULTS: Between 2003 and 2008, 228 patients underwent 238 procedures. Cerebral protection devices and self-expanding stents were placed in all patients. A total of 97 percutaneous interventions performed in 93 women were compared with 141 interventions in 135 men. Mean age in women was 71.8 +/- 9.2 years, in men was 72.2 +/- 9.1 years (P > .99); 44.3% of women and 34.7% of men had symptomatic disease (P = .14). Preoperative demographics and co-morbidities did not differ significantly between genders, with the exception of hypertension (83.0% of males vs 96.7% of females, P = .001), and history of coronary artery bypass grafting (31.8% of males vs 16.1% of females, P = .01). There were no significant differences seen in anatomic arterial characteristics, though there was a trend towards women having larger internal carotid to common carotid diameter ratios (0.65 vs 0.62) and more plaques isolated to the common carotid segment (9.5% vs 6.9%). There were no significant differences seen in overall 30 day peri-procedural stroke rate (2.1% in women and 4.2% in men, P = .48), death rate (0 % vs 0.7%, P > .99), or cardiac events (3.2% vs 0.7%, P = .3). The combined 30-day stroke, death, and MI rate was 5.7% for males compared to 5.4% for females (P > .99). There were no differences observed in the long-term survival, stroke-free survival, or restenosis between genders. CONCLUSION: Despite previous concerns over adverse outcomes in women undergoing carotid endarterectomy, from our data, carotid stenting appears to be a safe modality in women with equivalent outcomes when compared to men. PMID- 19216952 TI - Outcomes after carotid endarterectomy: is there a high-risk population? A National Surgical Quality Improvement Program report. AB - OBJECTIVE: Carotid endarterectomy (CEA) is the standard treatment of carotid stenosis for symptomatic and asymptomatic patients. Carotid angioplasty and stenting (CAS), however, has been proposed as alternative therapy for patients deemed at high-risk for CEA. This study examined 30-day adjudicated outcomes in a contemporary series of CEAs and assessed the validity of criteria used to define a potential high-risk patient population for CEA. METHODS: Patients undergoing isolated CEA in private sector hospitals between Jan 1, 2005, and Dec 31, 2006, were identified using the prospectively gathered National Surgical Quality Improvement Program database. The primary study end points were 30-day stroke and death rates. Demographic, preoperative, and intraoperative variables were examined using multivariate models to identify variables associated with the study end points. Variables used to define systemic "high-risk" patients in the Stenting and Angioplasty with Protection in Patients at High Risk for Endarterectomy (SAPPHIRE) study (active cardiac disease, severe chronic obstructive pulmonary disease, and octogenarian status) were examined individually and in composite fashion for association with study endpoints. RESULTS: Of the 3949 CEAs performed, 59% were in men, 30% were "high-risk" (19% age >80), and 43% had a previous neurologic event. The 30-day stroke rate was 1.6%, the death rate was 0.7%, and combined stroke/death rate was 2.2%. Multivariate analysis showed that intraoperative transfusion (odds ratio [OR], 5.95; 95% confidence interval [CI], 1.71-20.66; P = .005), prior major stroke (OR, 5.34; 95% CI, 2.96-9.64; P < .0001), shorter height (surrogate for small artery size; OR, 1.09; 95% CI, 1.02-1.16; P = .010), and increased anesthesia time (OR, 1.02; 95% CI, 1.00-1.03; P = .008) were predictive of stroke. Critical limb ischemia (OR, 12.72; 95% CI, 3.49-46.40; P < .0001) and poor functional status (OR, 7.05; 95% CI, 2.95-16.82; P < .0001) were independent correlates of death. Systemic high-risk variables, either combined or individually, did not increase risk of stroke or death on multivariate analysis. CONCLUSION: CEA is associated with favorable 30-day outcomes across a spectrum of patient comorbidity features including octogenarian status. Anatomic and technical features are the important predictors of perioperative stroke, whereas critical limb ischemia and poor functional status are important predictors of death for patients undergoing CEA. These data refute the concept that CAS is preferred for patients deemed high-risk by virtue of systemic comorbidities. PMID- 19216954 TI - Feasibility of simultaneous pre- and postfilter transcranial Doppler monitoring during carotid artery stenting. AB - OBJECTIVE: Carotid artery stenting (CAS) is emerging as an acceptable treatment alternative to surgery for patients with carotid artery stenosis. The major risk of CAS is cerebral embolization of plaque and thrombus causing stroke or asymptomatic brain infarction. Use of embolic protection devices (EPD) to trap emboli before they reach the brain is now standard practice in CAS. The pore size of the currently available filters is >100 microns and emboli smaller than the EPD pores can still reach the brain. While the use of EPD is widespread, little evidence exists of their in vivo efficacy in preventing distal embolization. Our aim was to quantify the number of emboli reaching the brain with the device in place. Therefore, the expected value of this report is in its description of a novel application of transcranial Doppler (TCD). Due to the limited number of cases, it is not intended to support the use of one EPD over another. METHODS: Six patients were monitored with ipsilateral simultaneous dual probe TCD during CAS. Two types of cerebral protection systems were evaluated: FilterWire EZ System (FW; Boston Scientific, Santa Clara, Calif) and GORE Neuro Protection System (NPS; W.L. Gore and Associates, Flagstaff, Ariz). By placing TCD probes both proximal and distal to the filterwire EPD, we quantified the microembolic signals before the EPD as well as those, which reached the intracranial circulation after the EPD. One probe was placed submandibularly to monitor the ICA (SICA), while another was placed transtemporally to monitor the middle and anterior cerebral artery (MCA + ACA). We compare the number of extracranial emboli prior to the EPD with the number of intracranial emboli after the EPD. RESULTS: Dual probe monitoring was successful during the five stages of the CAS: lesion crossing (LC), predilatation (PreD), stent placement (SP), postdilatation (PostD), and filter/device removal (FR/DR). Using FW during LC by probe 1 (SICA)/probe 2 (MCA + ACA): (18 [range, 15-22]/15 [range, 11-20]), PreD (111 [range, 101-121]/101 [range, 90-111]), SP (68 [range, 60-76]/42 [range, 30-53]), PostD (27 [range, 25-30]/24 [range, 22-27]), FR (0.3 [range, 0-1]/0.7 [range, 0 1]) average number of microembolic signals were detected. Using NPS during LC (1.7 [range, 0-3]/1 [range, 0-2]), PreD (0/1.7 [range, 0-4]), SP (0/0), PostD (0/0), DR (18 [range, 0-18]/6.7 [range, 1-13]) average number of microembolic signals were detected. CONCLUSION: EPD significantly reduces but does not eliminate the number of microemboli reaching the brain during carotid artery angioplasty and stenting. We propose monitoring of CAS with submandibular and transtemporal TCD probes to further evaluate the practice of distal embolization protection. Although our study is not powered to make any recommendations about EPDs, we believe that sequential dual probe TCD monitoring is a worthy tool with the potential to give vital information to assess the various devices and the techniques of utilization. PMID- 19216958 TI - Left renal vein transposition for nutcracker syndrome. AB - OBJECTIVE: Nutcracker syndrome, caused by compression of the left renal vein (LRV) between the superior mesenteric artery and the aorta, results in left renal and gonadal venous hypertension. Several treatment options have been described to relieve associated symptoms. The purpose of this study was to evaluate late results of LRV transposition and identify risk factors affecting outcomes. METHODS: Clinical data from 23 consecutive patients diagnosed with nutcracker syndrome from January 1996 to October 2007 were retrospectively reviewed. RESULTS: There were 10 males and 13 females (median age 22 years; range, 14-67) with radiologic evidence of LRV compression. On ultrasound evaluation (15/23 patients), the mean ratio of LRV peak systolic velocity measured at the site of compression and the renal hilum was 7.3 (range, 2.5-12). On venography (14/23 patients), the mean renocaval pressure gradient was 4 mm Hg (range, 2-6 mm Hg). Twelve patients with atypical abdominal pain (n = 4), hematuria (n = 5), and varicocele (n = 6) were managed expectantly. Eleven patients underwent LRV transposition through a transperitoneal exposure. Symptoms in these patients included left flank pain (n = 10), hematuria (n = 7), and varicocele (n = 3). In 2/11 patients, the LRV was found to be occluded at operation. There were no early postoperative complications. Most conservatively managed patients remained stable or improved over a mean follow-up period of 26 months (range, 0.2-59 months). Two patients were lost to follow-up at our institution and ultimately underwent intervention with LRV stenting and autotransplantation elsewhere. One patient was diagnosed with thin basement membrane disease on renal biopsy. Five patients with varicocele remained asymptomatic; 1 underwent local repair. Over a mean follow-up of 39 months (range, 0.13-144 months) in surgically managed patients, symptoms of flank pain and hematuria resolved or improved in 8/10 and 7/7, respectively. Varicoceles recurred in 2/3 patients in spite of resolution of flank pain. Both preoperatively occluded LRVs rethrombosed; one underwent thrombolysis with stenting, the other reimplantation of the left gonadal vein into the IVC. CONCLUSION: Evaluation of the clinical significance of radiologic LRV compression remains challenging, as does selection of patients for intervention. LRV transposition is a safe, effective procedure in selected patients with persistent, severe symptoms. Patients with progression to occlusion of the LRV should be considered for alternative therapeutic procedures. Varicoceles, in the setting of nutcracker syndrome, may need independent repair. PMID- 19216960 TI - Dose-response of compression therapy for chronic venous edema--higher pressures are associated with greater volume reduction: two randomized clinical studies. AB - BACKGROUND: Two phase II clinical studies used an experimental, multi-chambered compression device with different cuff pressure combinations in subjects with leg edema and chronic venous insufficiency. The objective of each study was to evaluate the safety and the relative effects of different cuff pressure combinations to determine if edema reduction was dose-dependent. METHODS: Each study enrolled adults with chronic (>or=6 weeks) venous edema corresponding to CEAP C(3)-C(5). The test device could apply different pressures at the foot, gaiter, mid-calf, and upper-calf. In the first study, the following six sustained pneumatic compression (SPC) profiles were applied for six hours each: 20, 30, and 40-mm Hg at the gaiter with graduated SPC (ie, lower pressures at the calf); and 20, 30, and 40-mm Hg at the gaiter with nongraduated SPC (ie, the same pressures at the calf). In the second study, the following three intermittent pneumatic compression (IPC) profiles were applied for two hours each: 40, 50, and 60-mm Hg at the gaiter with graduated IPC (ie, lower pressures at the calf). Each study included a baseline profile with no compression and two-day intervals between profiles. Leg volume was measured before and after compression using the water displacement method. RESULTS: A dose-response relationship was observed between increased SPC/IPC pressures and reduced limb edema. Limb volume was reduced most effectively with the highest pressures of 40-mm Hg nongraduated SPC and 60-mm Hg graduated IPC (136 mL and 87 mL, respectively); however, some subjects reported discomfort with these profiles. Limb volume was reduced by more than 100 mL with 30 to 40-mm Hg graduated SPC and by 69 mL with 50-mm Hg graduated IPC, and subjects rated these profiles as comfortable or very comfortable. Of the 28 study participants (12 SPC, 16 IPC), two subjects reported pain with 60-mm Hg IPC; no other adverse events were reported with SPC or IPC. CONCLUSION: Pneumatic compression was safe and well-tolerated, with a dose-response relationship between increased SPC/IPC pressures and reduced leg edema. To our knowledge, this is the first study to demonstrate a dose-relationship in compression therapy: higher pressures are associated with greater volume reduction in subjects with chronic venous edema. PMID- 19216961 TI - Posttraumatic edema of the lower extremities: evaluation of the lymphatic vessels with magnetic resonance lymphangiography. AB - OBJECTIVE: To assess for the first time the morphology of the lymphatic system in patients with posttraumatic edema of the lower extremities by magnetic resonance (MR) imaging using the interstitial lymphangiography technique MATERIALS AND METHODS: Six patients with posttraumatic edema in eight of their 12 lower extremities were examined by MR lymphangiography. Eighteen mL of gadoteridol and one mL of mepivacainhydrochloride 1% were subdivided into 10 portions and injected intracutaneously. MR imaging was performed with a 1.5-T system equipped with high-performance gradients. For MR lymphangiography, a 3D-spoiled gradient echo sequence was used. RESULTS: In five of the eight (63%) traumatized lower extremities, enlarged lymphatic vessels were detected, with the largest diameter measuring 5 mm. Additionally, a fast lymphatic outflow was observed in seven of the eight (88%) traumatized legs with enhancement of the inguinal lymph nodes already in the first image acquisition 15 minutes after contrast material injection. In two of the eight (25%) traumatized lower extremities, an extensive network of collateral lymphatic vessels was detected at the level of the calf. In both extremities, lymphatic collateralization involved not only the epifascial but also the subfascial lymphatic system. In one patient, who sustained a trauma of the left lower leg with tibial fracture, a small aneurysmatic widening of 7 mm could be detected at the middle level of the calf. CONCLUSION: MR lymphangiography is a safe and accurate minimal-invasive imaging modality for the evaluation of the lymphatic circulation in patients with posttraumatic edema of the lower extremities. If the extent of lymphatic damage is unclear at the initial clinical examination or requires a better definition for optimal therapeutic planning, MR lymphangiography is able to identify the anatomic and physiological derangements and to establish an objective baseline. PMID- 19216963 TI - Middle mesenteric artery arising from an inflammatory infrarenal aortic aneurysm. AB - We report of a patient with an inflammatory infrarenal aortic aneurysm with a diameter of 6.5 cm, a middle mesenteric artery (MMA) arising from the aneurysm, and a review of the literature. The patient underwent successful surgical treatment by using an interposition tube graft (Dacron graft, 18 mm) with replantation of the MMA. Reports about a MMA arising separately from the aorta are extremely rare, especially in combination with an infrarenal aortic aneurysm. In our case, it arose from the anterior aspect of the abdominal aorta, 6 cm below the superior mesenteric artery (SMA) and 1.2 cm above the inferior mesenteric artery (IMA). The MMA gave branches to the ileum and distal jejunum and supplied the iliocolic and middle colic artery branch as well as the left colic artery branch. It is of extreme clinical importance for the surgical procedure to have a detailed knowledge of the different anatomical variations and anomalies. PMID- 19216964 TI - Paraplegia as a symptom of failure after endovascular therapy of type B aortic dissection in Marfan syndrome. AB - This report describes successful treatment of an unusual case of concomitant paraplegia and type 1 endoleak during the early postoperative course of endovascular therapy of type B dissection in a patient with Marfan syndrome. PMID- 19216965 TI - Gluteopopliteal bypass for a compromised groin. AB - Extra-anatomic arterial reconstruction is indicated in patients with a compromised groin. Surgical options include obturator or transosseous bypass. We present a case of a patient with a necrotic radiation ulcer in the groin treated with a gluteopopliteal bypass. PMID- 19216966 TI - Extraluminal lipoma with common femoral vein obstruction: a cause of chronic venous insufficiency. AB - A 49-year-old man, with a misdiagnosis of common femoral vein deep vein thrombosis presented with advanced chronic venous insufficiency. Further imaging revealed a patent common femoral vein with augmentation that was compressed by an extrinsic mass. Exploration identified a lipoma that was extravascular and was resulting in venous outflow obstruction. Excision of the lipoma resulted in clinical improvement and ulcer healing. PMID- 19216967 TI - Erosion of elephant trunk Dacron graft limb by thoracic endograft causing acute aneurysm expansion. AB - We recently treated a patient in whom a Gore TAG thoracic endograft (W.L. Gore and Assoc, Flagstaff, Arix) had been used to repair a descending thoracic aneurysm as the second stage of a hybrid procedure. This patient had previously undergone repair of ascending and aortic arch aneurysms, with an elephant trunk graft limb placed in the descending thoracic aorta for subsequent repair of the descending thoracic aneurysm. Eight months after placement of the thoracic endograft, the patient presented with an acutely expanding and symptomatic thoracic aneurysm. The patient was operated on urgently. The proximal portion of the endograft had eroded into the previously placed Dacron elephant trunk limb. The proximal portion of the endograft was removed and was replaced with a Dacron graft. The management of this patient forms the basis of this report. PMID- 19216968 TI - Ruptured aneurysm of the ulnar artery in a woman with neurofibromatosis. AB - A 61-year-old woman with neurofibromatosis type 1 (Recklinghausen's disease) was referred for massive swelling of the right forearm, pain, increasing numbness, and impaired movement of the fingers. Angiography demonstrated a 13- x 11-mm aneurysm and a capped rupture of the ulnar artery. Because of the complicated soft-tissue condition, interventional treatment was indicated. Two 360 degrees coils were placed for embolization of the ruptured aneurysm. Arterial involvement in neurofibromatosis is a well known but infrequent occurrence. Stenotic lesions predominate. Aneurysmal defects are less common, and rupture of peripheral arteries is exceptional. PMID- 19216969 TI - Repair of multiple right renal artery aneurysms with associated fibromuscular dysplasia in a solitary kidney. PMID- 19216970 TI - Updated terminology of chronic venous disorders: the VEIN-TERM transatlantic interdisciplinary consensus document. AB - Non-uniform terminology in the world's venous literature has continued to pose a significant hindrance to the dissemination of knowledge regarding the management of chronic venous disorders. This VEIN-TERM consensus document was developed by a transatlantic interdisciplinary faculty of experts under the auspices of the American Venous Forum (AVF), the European Venous Forum (EVF), the International Union of Phlebology (IUP), the American College of Phlebology (ACP), and the International Union of Angiology (IUA). It provides recommendations for fundamental venous terminology, focusing on terms that were identified as creating interpretive problems, with the intent of promoting the use of a common scientific language in the investigation and management of chronic venous disorders. The VEIN-TERM consensus document is intended to augment previous transatlantic/international interdisciplinary efforts in standardizing venous nomenclature which are referenced in this article. PMID- 19216971 TI - Presidential address: Time to lead! PMID- 19216972 TI - Tips and tricks for avoiding access problems when using large sheath endografts. PMID- 19216973 TI - Changing demographics in patients with vascular disease. AB - The United States population with vascular disease has changed dramatically during the past 2 decades, with large increases in the proportion of Hispanic, African American, and other minority patients. Not only has the number of these patients increased, but the types and distribution of vascular disease in minority populations is also different from that encountered in non-Hispanic whites. Although genetic makeup accounts for some of these differences, access to vascular care is also an important determinant, with many minority patients presenting late in the course of the disease process. These factors create significant challenges for the vascular specialists caring for these patients. The vascular surgery workforce is composed of >90% white men and does not currently represent the changes in the population of patients with vascular disease. In addition, women with vascular disease comprise up to 50% of many vascular surgery practices. In many parts of the country, Hispanics and African Americans outnumber non-Hispanic whites with vascular disease. Yet, women and minority physicians are still significantly under-represented in the field of vascular surgery. This year's E. Stanley Crawford Critical Issues Forum at the Society for Vascular Surgery meeting addressed the disconnect between the vascular surgery workforce and the patients whom we serve. This article reviews the projected demographic changes in the population of the United States, which supports the need for training a vascular surgery workforce that is more diverse. This article also reviews the current status of minority and female representation in medical schools, surgical training programs, and vascular surgery programs in the United States. PMID- 19216976 TI - Regarding "Carotid endarterectomy with adjunctive carotid cephalad stenting: complimentary, not competitive techniques". PMID- 19216977 TI - Regarding "Midterm outcome after the distal revascularization and interval ligation (DRIL) procedure". PMID- 19216978 TI - Regarding "Carotid endarterectomy with adjunctive cephalad carotid stenting: Complementary, not competitive, techniques". PMID- 19216981 TI - The silver lining. PMID- 19216982 TI - Depression, anxiety and quality of life in suicide survivors: a comparison of close and distant relationships. AB - The study's purpose was to describe and compare depression, anxiety, and quality of life, by degree of relationship, between closely related and distantly related survivors (persons close to the suicide victim, or "suicide survivors"; N = 60) during the acute phase of bereavement (within 1 month of the death). The close relationship category included spouses, parents, children, and siblings, whereas the distant relationship category included in-laws, aunts/uncles, and nieces/nephews. Analysis of covariance examined differences between the two groups on the symptom measures. Results indicate that, after controlling for age and gender effects, closely related survivors had significantly higher mean levels of depression and anxiety and had lower levels of mental health quality of life. There were no statistically significant differences on the physical health quality of life subscale. PMID- 19216984 TI - An ethnographic approach to interpreting a mental illness photovoice exhibit. AB - The aim of this study was to explore the use of photovoice as an innovative methodology for understanding aspects of the mental illness experience. Applied ethnographic methods were used to examine 15 photographs and accompanying narratives from a mental illness photovoice exhibit. Analysis occurred within and across cases, resulting in individual case studies and a thematic description of the narratives. Suffering, stigma, and loss of identity were found at the center of this rendering of mental illness experience. The findings suggest that a photovoice project offers a useful lens from which to examine experiences associated with living with mental illness. PMID- 19216983 TI - Coping and schizophrenia: a re-analysis. AB - Standard notions of coping have not been particularly fruitful in the study of schizophrenia. However, facilitation of adaptive coping with serious mental illnesses such as schizophrenia is an important part of mental health care in general and of psychiatric nursing in particular. This study explored factors of coping and examined their relation with symptom severity and with quality of life of outpatients with schizophrenia. Data were analyzed from a previous cross sectional study, using the Ways of Coping Checklist, the Positive and Negative Syndrome Scale, and the Wisconsin Quality of Life Index. A principal component factor analysis was performed on the Ways of Coping Checklist scores, and the resulting six coping factors were then tested for correlations with Positive and Negative Syndrome Scale and Wisconsin Quality of Life Index scores. Factors conceptually linked with emotion-focused coping were more strongly associated with symptom severity and with quality of life than were factors conceptually linked with problem-focused coping. The emotion-focused versus problem-focused coping framework was only partly explanatory. It may be fruitful to study whether supportive counseling enhances beneficial factors conceptually linked with emotion-focused coping of individuals with schizophrenia. PMID- 19216985 TI - Behavioral change in patients with severe self-injurious behavior: a patient's perspective. AB - Semistructured interviews were conducted with 12 women who had successfully stopped self-injuring to gain an understanding of the process of stopping self injury. The data were analyzed based on the grounded theory method. The researchers found that the process of stopping self-injury consists of six phases. Connection was identified as key to all phases of the process. Nursing interventions should focus on forging a connection, encouraging people who self injure to develop a positive self-image and learn alternative behavior. PMID- 19216986 TI - Reducing stigma related to mental disorders: initiatives, interventions, and recommendations for nursing. AB - Twenty percent of Americans suffer from mental disorders, but most do not receive treatment. Stigma is an important barrier to mental health treatment and recovery. This article aims to summarize current national initiatives to reduce stigma, clarify the current knowledge of stigma-reducing interventions, and provide recommendations to nurses on implementing and investigating stigma reducing interventions. PMID- 19216987 TI - Intention to seek depression treatment in adolescent mothers and a comparison group of adolescent girls. AB - The aims of the study were (a) to determine the efficacy of the theory of reasoned action in predicting intention to seek depression treatment in adolescent mothers (n = 64) and a comparison group of adolescent girls (n = 65) and (b) to explore the role of empirically suggested variables (e.g., social support and current symptoms of depression) in increasing the explanatory power of the model. Findings between the two groups differed. Subjective norms, but not attitude, was a significant predictor of intention to seek depression treatment in adolescent mothers (F = 4.82, P = .00; R(2) = .14). In the comparison group of adolescent girls, both attitude and subjective norms predicted intention to seek depression treatment (F = 8.56, P = .00, R(2) = .22). The addition of depressive symptoms and social support increased the explanatory power of the model in the comparison group of adolescent girls (R(2) change = 0.10, P = .02) but resulted in the deletion of subjective norms from the regression equation. Implications for further research and practice are discussed. PMID- 19216988 TI - Depression among community-dwelling older adults in Korea: a prediction model of depression. AB - The purpose of this study was to test a hypothesized model for use in explaining the factors that influence depression among older adults dwelling in Ansan City in Korea. A cross-sectional community-based survey was done using face-to-face private interviews. A hypothesized model was tested using path analysis. Of the 32 hypothesized paths in the structural model, 13 were significant. Gender (beta = -.14, P = .24), income (beta = -.21, P < .00), education (beta = -.14, P = .10), activities of daily living (ADL; beta = -.29, P < .00), and somatic symptoms (beta = .29, P < .00) accounted for the incidence of depression (beta = .06, P < .00), representing 28% of the variance in depression. The fit of the model to the data was supported. PMID- 19216989 TI - Challenges in the treatment of factitious disorder: a case study. AB - Factitious disorder is difficult to diagnose and effectively treat. It is often met with intense emotion by both the care provider(s) and the client when suspected. However, if missed or untreated, it can become a chronic syndrome that is expensive and potentially dangerous. This article examines barriers to successful treatment of factitious disorder as manifested by a woman who received services from an urban community mental health center over a 7-year period for the management of multiple mental health issues. The unique role of the advanced practice psychiatric nurse is discussed through application of the nursing theory modeling and role modeling. PMID- 19216990 TI - Relationship between sense of belonging as connectedness and suicide in American Indians. AB - The incidence of suicide in American Indians is highest of all ethnic populations. M. Naegle and C. D'Avanzo (2001) assert that suicide is often correlated with primary depression and substance dependence. Prevalence rate for suicide within the American Indian population was 1.5 times the national rate in 2001. The sociohistorical impact of sense of belonging to suicide within this population has not been explored in previous research (C. Strickland, E. Walsh, & M. Cooper, 2006). The findings support that sense of belonging has a negative association with suicidal ideation (r = -.203, P < or = .01) and may buffer the development of depressive symptoms. PMID- 19216991 TI - Use of antidepressants and complementary and alternative medicine among outpatients with depression in Taiwan. AB - Understanding use of complementary and alternative medicine (CAM) and antidepressants during patient's recovery for depression is important to enhance their psychological functioning and promote positive health care outcomes. This study investigated the use of antidepressant treatment and CAM in outpatients with depression 1 month after discharge from psychiatric hospitalization. Telephone surveys were conducted. Of the 201 participants, 50.2% reported using CAM 1 month after discharge. Nearly 41% (n = 82) used a combination of both antidepressant treatment and CAM after hospital discharge. The most commonly used CAMs were spiritual healing, relaxation techniques, and herbal medicine. Symptom relief was the top reason participants used CAM for depression. A number of factors were associated with higher odds of using CAM. CAM is often considered as a valued component of holistic care plan after discharge. It is important for nurses to identify the CAM approaches patients are using so that this information can assist in the education of patients and family about the benefits of contemporary treatments for depression, possible interactions when combining treatments, and the CAM and conventional medicine that can be helpful to relieve depression symptoms and psychological distress. PMID- 19216992 TI - A convergence of formative events to guide mental health nursing research. PMID- 19216993 TI - Nurses need to debate involuntary outpatient treatment. PMID- 19216994 TI - Editorial: Silkworm genome. PMID- 19216995 TI - Mapping of sex-linked genes onto the genome sequence using various aberrations of the Z chromosome in Bombyx mori. AB - Many strains of Bombyx mori carry chromosomal aberrations, and they are useful resources for integration between phenotypes and genomic sequences. We compared the molecular structures of three kinds of Z chromosomes, i.e., two strains with chromosome deletions and one strain with translocation involving the Z chromosome. Using polymerase chain reaction markers, we showed that: (1) the Z(1) chromosome lacks more than 6Mb, including the proximal end; (2) the Z(Vg) chromosome lacks 1.5Mb in the interstitial portion; and (3) the +(od)p(Sa)+(p)W carries a 0.6-Mb Z-derived fragment surrounding the +(od) gene. The breakpoint junctions of these deletions and a translocation were precisely determined. Through deletion mapping, we narrowed down the regions where distinct oily (od), vestigial (Vg), and muscle dystrophy (Md) are located and identified a candidate gene for od. A retroposon-mediated deletion in BmBLOS2--the Bombyx gene homologous to human "biogenesis of lysosome-related organelles complex-1, subunit 2''--was detected in the od mutant. Although the genes responsible for Vg and Md were not definitively identified, we propose the candidate genes on the basis of their locations and phenotypes. PMID- 19216996 TI - Is the metabolic syndrome a discrete diagnostic category or the end of a continuum? Taxometric evidence for dimensionality in the National Health and Nutrition Examination Survey 1999-2004. AB - PURPOSE: To determine whether the metabolic syndrome is a discrete diagnostic category or the end of a continuum. METHODS: The study sample consisted of 3959 individuals from the National Health and Nutrition Examination Survey 1999-2004. The taxometric methods of mean above minus mean below a cut (MAMBAC) and maximum covariance (MAXCOV) were used. The primary numerical outcome was the comparative curve fit index (CCFI). RESULTS: For men, the CCFI from MAMBAC and MAXCOV analyses were 0.290 and 0.260, respectively. Likewise, for women, the CCFI from MAMBAC and MAXCOV analyses were 0.317 and 0.311, respectively. CONCLUSIONS: The results indicate that the metabolic syndrome is continuously distributed in nature, rather than a discrete diagnostic category. PMID- 19216997 TI - An international case-control study of maternal diet during pregnancy and childhood brain tumor risk: a histology-specific analysis by food group. AB - PURPOSE: Maternal dietary data from an international collaborative case-control study on childhood brain tumors were used to evaluate associations between histology-specific risk and consumption of specific food groups during pregnancy. METHODS: Nine study centers from seven countries contributed 1218 cases and 2223 controls. Most cases were diagnosed between 1982 and 1992 and ranged in age from 0 to 19 years. Dietary consumption was measured as average grams per day. RESULTS: Foods generally associated with increased risk were cured meats, eggs/dairy, and oil products; foods generally associated with decreased risk were yellow-orange vegetables, fresh fish, and grains. The cured meat association was specific to astrocytomas (odds ratio [OR] range=1.8-2.5 across astrocytoma subtypes for 4th vs. 1st quartile of consumption, p trends or=3 vs. <3) outcome. RESULTS: We found strong inverse associations between NSES and AL for black subjects, after adjusting for age, sex, U.S. birth, urban location, and individual SES. These associations were weaker and less consistent for Mexican Americans and whites. CONCLUSIONS: Our results indicate that living in low NSES neighborhoods is most strongly associated with greater cumulative biological risk profiles in the black U.S. population. PMID- 19217004 TI - Screening for serious mental illness in the National Survey on Drug Use and Health (NSDUH). PMID- 19217006 TI - Why are so many epidemiology associations inflated or wrong? Does poorly conducted animal research suggest implausible hypotheses? AB - There is growing concern among epidemiologists that most discovered associations are either inflated or false. The reasons for this concern have focused on methodological issues in the conduct and publication of epidemiologic research. This commentary suggests that another reason for discrepant findings may be that animal research is producing implausible hypotheses. Many animal studies are methodologically weak, and the animal literature is not systematically reviewed and synthesized. Moreover, most bodies of animal literature may be so heterogeneous that they can be used selectively to support the plausibility of almost any epidemiology study result. Epidemiologists themselves also do not consistently conduct systematic reviews of bodies of biological evidence which might point to sources of bias in an evidence base. Animal research will likely continue to provide the biological basis for epidemiological investigation, but substantial improvement is needed in how it is conducted and synthesized to improve the predictability of animal studies for the human condition. PMID- 19217005 TI - U.S. burden of disease--past, present and future. AB - PURPOSE: To review the history and challenges of "burden of disease" studies, how these are dependent on robust epidemiologic data as well as complex conceptual constructions, and to identify the public health policy issues these studies can most usefully inform. METHODS: The emergence of the concept of the "burden of disease" in the public health literature is reviewed, with a focus on the results of an analysis of data from the United States that used the methodology presented in the Global Burden of Disease Study. RESULTS: The systematic analysis of public health mortality data to identify major health problems was conducted by Graunt in 16th-century London. He found that many of the predominant sources of mortality were not the focus of public attention. Today, despite refinements in epidemiologic measurement methods designed to capture the impact of non-fatal health conditions, there are similar incongruities between the major public health problems and expenditures on prevention interventions. CONCLUSIONS: Controversies surrounding the interpretation of "burden of disease" studies are not new. Particularly in developed countries, these studies appear more useful for setting research priorities rather than allocating resources to support prevention efforts. Such investigations are not possible without ongoing support for systematic collection and analysis of descriptive epidemiologic data. PMID- 19217007 TI - Retrospective comparison of surgical treatment modalities in 100 patients with oral submucous fibrosis. AB - BACKGROUND AND OBJECTIVE: Oral submucous fibrosis has been a scourge of southeastern Asia and its residents since time immemorial. Scores of medicinal agents, singly and in combination, have been tried with not very encouraging results. In this study, therefore, we have restricted ourselves to different surgical modalities in the management of this condition and have tried to lay down indications of each surgical procedure. STUDY DESIGN: A total of 100 patients of oral submucous fibrosis were included in this study and randomly allocated to different surgical groups, 25 patients per group. After excision of fibrous bands, group I had buccal fat pad graft, group II had tongue flap, group III had nasolabial fold flap, and group IV had split skin graft for correction of mucosal defect created after incising the fibrous bands. RESULTS: Mean preoperative mouth opening was 14.82 (SD 4.38) mm and ranged between 4.00 and 25.00 mm. Statistically there was no significant difference among the 4 groups (P = .996). Mean postoperative mouth opening at 1 week was 35.79 (3.53) mm, ranging between 24.00 and 42.00 mm. Mean postoperative mouth opening at 1 month in group I was 36.36 (2.64) mm, in group II 35.36 (29) mm, in group III 35.64 (2.94) mm, and in group IV 35.80 (3.24) mm. Total score for pain, esthetics, and function at 1 month after surgery was highest (11.29) in group I, indicating better results. CONCLUSION: The results were very encouraging, and we were able to lay down specific indications for each procedure. However, we believe buccal fat pad rotation is superior to other procedures, because it offers ease of surgery, can be performed under local anesthesia as a day care procedure, shows little postoperative morbidity, and has good patient acceptance, and there appear to be no contraindications to its use. PMID- 19217008 TI - Effect on pocket depth and attachment level of manual versus ultrasonic scaling of lower second molars following lower third molar extraction: a randomized controlled trial. AB - OBJECTIVE: The objective of this study was to compare the effect on periodontal health of ultrasonic versus manual scaling of the lower second molar (L2M) root surface following surgical extraction of mesioangular lower third molars (L3M). STUDY DESIGN: Randomized single-blind clinical trial to compare the effectiveness of ultrasonic scaling of the L2M (experimental group) and manual scaling and root planing (control group) after surgical extraction of the corresponding L3M. RESULTS: Forty-three subjects were enrolled, 69.8% completing the study. The postoperative bony defect on the distal surface of the L2M was significantly reduced for both treatment groups (P = .001). Despite a higher reduction of intrabony defect being observed in the ultrasonic group, the difference was not significant (P = .17) at 6 months. Furthermore, there was also no statistically significant correlation (P > .05) between the postoperative pocket depth and the treatment group (ultrasonic or manual). CONCLUSIONS: Pocket depth and depth of the intrabony defect were similar in both experimental and control groups. PMID- 19217009 TI - Evaluation of peri-implant tissue response according to the presence of keratinized mucosa. AB - OBJECTIVES: The purpose of this study was to evaluate the responses of peri implant tissue in the presence of keratinized mucosa. STUDY DESIGN: A total of 276 implants were placed in 100 patients. From the time of implant placement, the average follow-up observation period was 13 months. The width of keratinized mucosa was compared and evaluated through the gingival inflammation index (GI), plaque index (PI), the pocket depth, mucosal recession, and marginal bone resorption. RESULTS: The GI, PI, and pocket depth in the presence or absence of the keratinized gingiva did not show statistically significant differences. However, mucosal recession and marginal bone resorption experienced statistically significant increases in the group of deficient keratinized mucosa. Based on implant surface treatments, the width of keratinized gingiva and crestal bone loss did not show a significant difference. CONCLUSION: In cases with insufficient keratinized gingiva in the vicinity of implants, the insufficiency does not necessarily mediate adverse effects on the hygiene management and soft tissue health condition. Nonetheless, the risk of the increase of gingival recession and the crestal bone loss is present. Therefore, it is thought that from the aspect of long-term maintenance and management, as well as for the area requiring esthetics, the presence of an appropriate amount of keratinized gingiva is required. PMID- 19217010 TI - Oral anomalies associated with the oculocerebrorenal syndrome of Lowe: case report with multiple unerupted teeth and pericoronal radiolucencies. AB - The oculocerebrorenal syndrome of Lowe (OCRL) is a rare X-linked recessive disorder, chiefly characterized by ocular involvement, mental retardation, and kidney disease. A literature review is provided, detailing the diversity of oral anomalies associated with the OCRL syndrome. Reported abnormalities include delayed tooth eruption, odontogenic cyst formation, and constricted dental arches. In addition, we present an unusual case of an 18-year-old male affected with the OCRL syndrome and fetal alcohol syndrome. The oral radiographic examination was significant for multiple impacted permanent teeth, many with pericoronal radiolucencies, and an underdeveloped mandible. PMID- 19217011 TI - Panoramic radiographic findings as predictors of mandibular nerve exposure following third molar extraction: digital versus conventional radiographic techniques. AB - OBJECTIVE: The aim was to compare digital and conventional panoramic imaging techniques for identifying high-risk radiographic markers associated with mandibular nerve (MN) injury after mandibular third molar (M3) removal. STUDY DESIGN: The study used a retrospective cohort model. The predictor variable was the presence or absence of radiographic signs associated with MN exposure during M3 removal. The outcome variable was MN exposure. Intraexaminer variability was estimated using a kappa statistic. Logistic regression modeling was used to measure the association between radiographic signs and MN exposure and determine if imaging technique modified that association. The level of statistical significance was set at P < .05. RESULTS: The sample was composed of 571 patients having 1,017 mandibular M3s removed. The MN was visualized in 66 extraction sites (6.5%). In the adjusted logistic model, radiographic signs were statistically associated with MN exposure (P < .001) and imaging technique was a statistically insignificant effect modifying variable (P = .4). CONCLUSION: The results of this study suggest that imaging technique does not modify the relationship between high-risk panoramic radiographic signs and MN exposure. PMID- 19217012 TI - Effect of oral appliances on genioglossus muscle tonicity seen with diffusion tensor imaging: a pilot study. AB - OBJECTIVE: The purpose of this study was to examine whether the diffusion tensor imaging (DTI) technique can be used as a modality to represent the structural deformation in the in vivo genioglossus (GG) muscle fibers with oral appliances (OAs). STUDY DESIGN: Three healthy subjects were recruited for the pilot study. A custom-made OA, which is modified from a tongue retaining device (TRD), was constructed for each subject before the DTI acquisitions. Recordings were made with and without OAs to compare the GG muscle fiber deformation. RESULT: DTI provided good resolution of tongue muscle fibers in vivo and successful isolation of each muscle fiber bundle. In particular, the GG muscle fiber deformation due to OAs was clearly visualized. CONCLUSIONS: This DTI technique may be used not only to identify the individual myoarchitecture, but also to assess muscle fiber deformations in vivo, such as constriction, dilatation, and rotation with OAs. Clinical studies for OSA patients will be the next step. PMID- 19217013 TI - Advances in hematologic stem cell transplant: an update for oral health care providers. AB - Oral supportive care is critical in the management of patients receiving hematopoietic cell transplantation (HCT). Advances in HCT, such as the use of stem cells isolated from peripheral blood instead of bone marrow, have resulted in more rapid engraftment and thus a shorter duration of pancytopenia. Reduced intensity conditioning regimens, associated with less toxicity, make HCT available to older patients and patients with comorbidities. These new developments have led to increased transplant rates and an altered spectrum of therapy-related complications, such as mucositis, and to shifts in the prevalence and pattern of occurrence of infections and graft-versus-host disease. The purpose of this paper is to review the main principles of HCT and to update dental providers on new technologies being applied to transplantation that may influence oral complications and oral care. PMID- 19217014 TI - Dental implants in a patient with Paget disease under bisphosphonate treatment: a case report. AB - Paget disease (PD) is an idiopathic disorder characterized by progressive enlargement and deformity of bones, resulting in structural weakness that may be unfavorable for the placement of osseointegrated dental implants. Currently, bisphosphonates are administered for the treatment of PD. However, the use of bisphosphonates has been associated with the onset of osteonecrosis of the jaws. This report deals with a case of a 64-year-old caucasian woman diagnosed with polyostotic PD 17 years earlier who had been treated with bisphosphonates for 7 years. The patient was referred for implant treatment, and after initial placement of 6 fixtures, the postoperative course was uneventful and no complications were registered during a follow-up period of 4 years. Within the limitations of this case, we report that dental implants can be successfully placed and loaded in patients suffering from PD without discontinuing the treatment with bisphosphonates, as long as the patient is not complaining from any additional unfavorable conditions. To the best of our knowledge, this is a unique case describing the successful placement of dental implants in a patient with PD treated with bisphosphonates. Nevertheless, more studies would be needed to optimize the clinical guidelines for the treatment of these patients. PMID- 19217015 TI - Ror2 expression in squamous cell carcinoma and epithelial dysplasia of the oral cavity. AB - In this study, the expressions of Ror2 in the normal mucosa, the epithelium dysplasia, and squamous cell carcinoma (SCC) of the oral cavity were investigated, and possible differences in the expression patterns of Ror2 and of p53, Ki67, or PCNA were examined. In Western blotting analyses, Ror2 expression in oral cancer was significantly higher than that in the normal oral mucosa. Immunohistochemically, Ror2 was localized on the plasmalemma and in the rough endoplasmic reticulum (rER). The tissue area with an Ror2-positive expression tended to differ from the area with a positive expression of p53, ki67, or PCNA, and the number of cells with an Ror2 expression tended to increase as the degree of malignancy rose in the epithelial tissues. These results suggest that Ror2 was not related to cell proliferation, but rather associated with cell polarity and cell motility, and that it was also closely associated with the degree of malignancy in oral epithelial tissue. PMID- 19217016 TI - Comparative radiographic and histological analyses of periapical lesion development. AB - OBJECTIVE: The present study evaluated the dynamics of the development of periapical lesions. STUDY DESIGN: Root canals from dogs' teeth were exposed to the oral environment, and then sealed for 7 days (Group A), 15 days (Group B), 30 days (Group C), and 60 days (Group D). After each experimental period, radiographs were taken to detect periapical bone resorption. In addition, histological sections from the periapical region were prepared. The radiographic and histological results were analyzed by ANOVA and Tukey's, Wilcoxon, and Pearson's tests. Significance level was set at 5%. RESULTS: Lesions were radiographically visible at 15 and 30 days, and had similar size at these periods (P > .05). At 60 days, lesions were larger than in the other periods (P < .05). Bone resorption was detected histologically at 7 days. The greatest values of bone resorption were observed at the 30- and 60-day periods (P < .05). The results of the methods of evaluation were similar only at the 30-day period. There was no correlation between the radiographic and histological results. CONCLUSIONS: Periapical radiography did not provide detection of periapical lesion in its initial stages. The periapical lesions became more evident radiographically when the bone resorption area increased. PMID- 19217017 TI - The European Journal of Cardiovascular Nursing. PMID- 19217018 TI - Gels and microgels for nanotechnological applications. AB - In recent years, "smart" materials have been the focus of considerable interest, from both fundamental and applied perspectives. Polymer gels are within this category; they respond to specific environmental stimuli by changing their size. Thus, the internal structure, the refractive index, and the mechanical properties of the polymer network change. They are considered super absorbent materials, as they can absorb solvent up to several hundred times their own weight. They respond rapidly to local environmental variations, an important fact in device miniaturization and microsensor developments. As size changes are accompanied by changes in internal dimensions, microgels have found application as carriers of therapeutic drugs and as diagnostic agents. They have also been used as microreactors, optically active materials, for template synthesis of nanoparticles or fabrication of artificial muscle. In this paper we review a set of application based on the special features associated to this systems. Basic concepts on the physical-chemistry of gel swelling is first described, followed by different applications covering drug delivery, composite materials using polymer gels to modulate optical or magnetic and electrical properties, molecular imprinting, gel-based biosensors and polymer sensors and actuators used in the field of artificial muscles. PMID- 19217019 TI - Effects of ranibizumab in patients with subfoveal choroidal neovascularization attributable to age-related macular degeneration. AB - PURPOSE: To demonstrate not only prevention of vision loss but also improvement in best-corrected visual acuity (BCVA) after treatment with ranibizumab on a variable-dosing regimen over 24 months in patients with age-related macular degeneration (AMD). DESIGN: Interventional case series. METHODS: SETTING: Institutional. STUDY POPULATION: One hundred and thirty-eight eyes of 138 patients treated intravitreally with 0.5 mg ranibizumab (Lucentis; Novartis, Basel, Switzerland). Age above 50 years, BCVA 0.2 to 1.2 logarithm of the minimal angle of resolution (logMAR), primary or recurrent subfoveal choroidal neovascularization (CNV) secondary to AMD. OBSERVATION PROCEDURES: After single initial treatment, monthly follow-up examination. Retreatment in case of one of the following: sign of subretinal fluid or intraretinal edema, increase in central retinal thickness (CRT) on optical coherence tomography (OCT), active CNV on fluorescein angiography, increase of metamorphopsia, and loss of BCVA > 5 letters on Early Treatment Diabetic Retinopathy Study (ETDRS) chart. MAIN OUTCOME MEASURES: Compared with baseline: proportion of eyes gaining > or = 15 letters, proportion of eyes losing or gaining < 15 letters, change in CRT. RESULTS: After 24 months, 30% of eyes gained > or = 15 letters. After 24 months, 55% of eyes lost or gained < 15 letters. Mean CRT of 386 +/- 145 microm at baseline was significantly reduced to 211 +/- 39 microm after 24 months (P = .036). Mean injection number per patient was 5.6 +/- 2.9 and 4.3 +/- 3.8 from baseline to month 12 and month 12 to 24, respectively. CONCLUSION: Intravitreal ranibizumab on a variable-dosing regimen was effective in significantly increasing mean BVCA and reducing CRT. This beneficial outcome was achieved with a low-rate of mild ocular adverse effects among our patients. PMID- 19217020 TI - Severe Legionella pneumophila pneumonia following infliximab therapy in a patient with Crohn's disease. AB - BACKGROUND: Immunosuppressive therapy with anti-TNF-alpha antibodies is effective in patients with inflammatory bowel disease (IBD). However, there is an increased risk for infections associated with this therapy. METHODS: Here, we report the case of a 58-year-old patient with Crohn's disease (CD) treated with steroids and azathioprine who developed severe Legionella pneumophila pneumonia after 3 infusions of infliximab. The patient presented at our IBD department with severe active CD complicated by inflammatory small bowel stenoses and entero-enteral fistulas despite long-term high-dose steroid therapy. To achieve steroid tapering and control of disease activity, immunosuppressive therapy with azathioprine was initiated. Due to persistent symptoms, infusion therapy with the anti-TNF-alpha antibody infliximab was started, subsequently leading to significant clinical improvement. However, after the third infliximab infusion the patient was hospitalized with fever, severe fatigue, and syncope. RESULTS: Laboratory findings and chest X-ray revealed left-sided pneumonia; cultural analysis showed L. pneumophila serogroup 1 leading to respiratory insufficiency, which required mechanical ventilation for 2 weeks in the intensive care unit. After discontinuation of all immunosuppressive agents and immediate antibiotic therapy the patient recovered completely. CONCLUSIONS: To our knowledge, this is the third case of L. pneumophila pneumonia in an IBD patient treated with infliximab. Similar to other published cases, concomitant treatment of immunosuppressives and anti-TNF agents is a major risk factor for the development of L. pneumophila infection, which should be ruled out in all cases of pneumonia in patients with such a therapeutic regimen. Appropriate prevention strategies should be provided in these patients. PMID- 19217021 TI - Likelihood ratio tests for maternal and fetal genetic effects on obstetric complications. AB - It is well recognized that both maternal and fetal genes could contribute to susceptibility for obstetric complications. Logistic regression models are usually adopted to model the separate or joint action of maternal and fetal loci with case-control data. The standard likelihood ratio tests (LRTs) can be used to test the significance of appropriate odds ratio parameters. This method, although simple to implement, fails to exploit a unique feature of genetic epidemiology studies of obstetric complications. Specifically, it does not take into consideration the correlation between the maternal and offspring genomes. We propose novel LRT that take advantage of this information by incorporating the fact that half of a child's genome is inherited from the mother. Our methods have substantially improved power for detecting marginal, main, and interactive maternal and fetal genotype effects, as evidenced by results from extensive simulation studies. We demonstrate our new methods by applying them to the analysis of data from a pilot study of preeclampsia. PMID- 19217023 TI - Mitotic catastrophe as a prestage to necrosis in mouse liver cells treated with Helicobacter pullorum sonicates. AB - Helicobacter pullorum infections have been associated with several enterohepatic diseases, but the mechanism of action is currently undefined. The present study was therefore set up to investigate possible cytotoxic effects of this pathogen on liver cells. A mouse hepatic cell line was exposed to H. pullorum sonicate and cytotoxicity was observed for all isolates after incubation for 72 h. Features characteristic for mitotic catastrophe characterized by chromatin condensation, formation of multinuclear distended cells and micronucleation, were recorded. In addition, intranuclear pseudoinclusions were seen in sonicate-treated cells. Finally, cells exposed to sonicate eventually underwent cell death with the morphological features of necrosis, which occurred without activation of caspase 3. The toxic factor involved in the cytotoxic activity proved to be soluble, trypsin-sensitive and stable at 56 degrees C and at -70 degrees C with a molecular weight to be over 50 kDa. The current study shows for the first time that H. pullorum causes mitotic catastrophe resulting in primary necrosis in mouse hepatocytes. PMID- 19217022 TI - The sumLINK statistic for genetic linkage analysis in the presence of heterogeneity. AB - We present the "sumLINK" statistic--the sum of multipoint LOD scores for the subset of pedigrees with nominally significant linkage evidence at a given locus- as an alternative to common methods to identify susceptibility loci in the presence of heterogeneity. We also suggest the "sumLOD" statistic (the sum of positive multipoint LOD scores) as a companion to the sumLINK. sumLINK analysis identifies genetic regions of extreme consistency across pedigrees without regard to negative evidence from unlinked or uninformative pedigrees. Significance is determined by an innovative permutation procedure based on genome shuffling that randomizes linkage information across pedigrees. This procedure for generating the empirical null distribution may be useful for other linkage-based statistics as well. Using 500 genome-wide analyses of simulated null data, we show that the genome shuffling procedure results in the correct type 1 error rates for both the sumLINK and sumLOD. The power of the statistics was tested using 100 sets of simulated genome-wide data from the alternative hypothesis from GAW13. Finally, we illustrate the statistics in an analysis of 190 aggressive prostate cancer pedigrees from the International Consortium for Prostate Cancer Genetics, where we identified a new susceptibility locus. We propose that the sumLINK and sumLOD are ideal for collaborative projects and meta-analyses, as they do not require any sharing of identifiable data between contributing institutions. Further, loci identified with the sumLINK have good potential for gene localization via statistical recombinant mapping, as, by definition, several linked pedigrees contribute to each peak. PMID- 19217024 TI - A new measure of the effective number of tests, a practical tool for comparing families of non-independent significance tests. AB - p-Values from tests of significance can be combined using the Sidak correction (or the closely related Bonferroni correction) or Fisher's method, but both these methods require that the p-values combined be independent when all null hypotheses tested are true. In this paper adjustments to these methods are proposed, using a new eigenvalue-based measure of the effective number of independent tests to which the actual tests performed are equivalent, and are compared with adjustments proposed by previous authors. The adjusted methods are evaluated using a sample of 726 Alzheimer's disease (AD) cases and 707 group matched controls, genotyped at 84,975 single-nucleotide polymorphism loci in 2,000 randomly chosen genes. The tests for genetic association with AD at loci within each gene are combined. The number of loci tested per gene varies from 2 to 994. The adjusted combined p-values agree well with the significance of the combined p-values determined empirically by random permutation of the data (Sidak correction: r=0.990; Fisher's method: r=0.994). This indicates that the combined p-values can be used to assess the relative strength of evidence for association of these genes with AD. The adjustment proposed here is a refinement of that of Nyholt ([2004] Am. J. Hum. Genet. 74:765-769), giving improved agreement with the results of random permutation. The improvement obtained is similar to that given by the refinement proposed by Li and Ji ([2005] Heredity 95:221-227). It is concluded that the concept of an effective number of tests is a valid approximation that allows p-values to be combined in a highly informative way. PMID- 19217025 TI - Positron emission tomography imaging of amphetamine-induced dopamine release in the human cortex: a comparative evaluation of the high affinity dopamine D2/3 radiotracers [11C]FLB 457 and [11C]fallypride. AB - The use of PET and SPECT endogenous competition binding techniques has contributed to the understanding of the role of dopamine in several neuropsychiatric disorders. An important limitation of these imaging studies is the fact that measurements of acute changes in synaptic dopamine have been restricted to the striatum. The ligands previously used, such as [(11)C]raclopride and [(123)I]IBZM, do not provide sufficient signal to noise ratio to quantify D(2) receptors in extrastriatal areas, such as cortex, where the concentration of D(2) receptors is much lower than in the striatum. Given the importance of cortical DA function in cognition, a method to measure cortical dopamine function in humans would be highly desirable. The goal of this study was to compare the ability of two high affinity DA D(2) radioligands [(11)C]FLB 457 and [(11)C]fallypride to measure amphetamine-induced changes in DA transmission in the human cortex. D(2) receptor availability was measured in the cortical regions of interest with PET in 12 healthy volunteers under control and postamphetamine conditions (0.5 mg kg(-1), oral), using both [(11)C]FLB 457 and [(11)C]fallypride (four scans per subjects). Kinetic modeling with an arterial input function was used to derive the binding potential (BP(ND)) in eight cortical regions. Under controlled conditions, [(11)C]FLB 457 BP(ND) was 30-70% higher compared with [(11)C]fallypride BP(ND) in cortical regions. Amphetamine induced DA release led to a significant decrease in [(11)C]FLB 457 BP(ND) in five out the eight cortical regions evaluated. In contrast, no significant decrease in [(11)C]fallypride BP(ND) was detected in cortex following amphetamine. The difference between [(11)C]FLB 457 and [(11)C]fallypride ability to detect changes in the cortical D(2) receptor availability following amphetamine is related to the higher signal to noise ratio provided by [(11)C]FLB 457. These findings suggest that [(11)C]FLB 457 is superior to [(11)C]fallypride for measurement of changes in cortical synaptic dopamine. PMID- 19217026 TI - Pharmacological characterization of 2-methoxy-N-propylnorapomorphine's interactions with D2 and D3 dopamine receptors. AB - Dopaminergic signaling pathways have been extensively investigated using PET imaging, primarily with antagonist radioligands of D(2) and D(3) dopamine receptors (DARs). Recently, agonist radioligands of D(2)/D(3) DARs have begun to be developed and employed. One such agonist is (R)-2-(11)CH(3)O-N-n propylnorapomorphine (MNPA). Here, we perform a pharmacological characterization of MNPA using recombinant D(2) and D(3) DARs expressed in HEK293 cells. MNPA was found to robustly inhibit forskolin-stimulated cAMP accumulation to the same extent as dopamine in D(2) or D(3) DAR-transfected cells, indicating that it is a full agonist at both receptors. MNPA is approximately 50-fold more potent than dopamine at the D(2) DAR, but equally potent as dopamine at the D(3) DAR. MNPA competition binding curves in membrane preparations expressing D(2) DARs revealed two binding states of high and low-affinity. In the presence of GTP, only one binding state of low affinity was observed. Direct saturation binding assays using [(3)H]MNPA revealed similar results as with the competition experiments leading to the conclusion that MNPA binds to the D(2) DAR in an agonist-specific fashion. In contrast to membrane preparations, using intact cell binding assays, only one site of low affinity was observed for MNPA and other agonists binding to the D(2) DAR. MNPA was also found to induce D(2) DAR internalization to an even greater extent than dopamine as determined using both cell surface receptor binding assays and confocal fluorescence microscopy. Taken together, our data indicate that the PET tracer, MNPA, is a full and potent agonist at both D(2) and D(3) receptors. PMID- 19217028 TI - The significance of endocervical cells and metaplastic squamous cells in liquid based cervical cytology. AB - We conducted a retrospective study to investigate whether the presence or absence of endocervical cells (EC) and metaplastic squamous cells (MSC) was associated with the detection of squamous intraepithelial lesions in liquid-based cervical cytology. 90,376 cases of liquid-based cervical cytology smears received in 2006 were included in the study. Low-grade (LSIL) and high-grade squamous intraepithelial lesions (HSIL) were classified according to the Bethesda system (2001). The rates of detecting LSIL and HSIL in smears with and without EC and/or MSC were determined. There were 1,540 LSIL and 396 HSIL. The ratio of HSIL/NILM (no intraepithelial lesion or malignancy) was 0.0022 in smears without EC or MSC, 0.0040 in smears with EC only, 0.0044 in smears with MSC only, and 0.0056 in smears with both EC and MSC present. Compared with smears without EC or MSC, this ratio was significantly higher (P < 0.05) when either EC or MSC was present. Compared with smears with EC only, the ratio was also significantly higher when both EC and MSC were present (P < 0.05). On the other hand, the presence or absence of EC had no effect on the detection rate of LSIL (0.0191 for both groups), while the presence of MSC was actually associated with lower detection rate of LSIL (0.0153, P < 0.05). The presence of endocervical and metaplastic cells was associated with higher detection rates of HSIL. MSC was associated with lower detection or LSIL. PMID- 19217027 TI - Increased dopamine D2High receptors in rats reared in social isolation. AB - Postweaning social isolation in the rat induces lasting alterations that parallel several of the core symptoms seen in human schizophrenics, including hyperreactivity to novel environments, cognitive impairment, and deficits in sensorimotor gating. The current study determined whether these changes are accompanied by any elevation in the proportion of striatal dopamine receptors in the functional high affinity state (D(2) (High)), as observed in other preclinical models of psychosis. Male Lister hooded rats (20-24 days) were housed in groups of three or alone. On Day 36 postweaning locomotor activity was monitored for 60 min in a novel arena, and on Day 37 novel object discrimination was assessed using a 2 h intertrial interval. Three days later striata were collected, homogenized, washed three times to remove endogenous dopamine, and the proportion of D(2) (High) determined by competition between dopamine and 2.27 nM [(3)H]domperidone. Isolates were significantly more active than group housed controls for both ambulation and rears. Although both groups exhibited comparable levels of familiarization trial object exploration, group housed animals were able to discriminate between novel and familiar objects during the choice trial while isolates were not. Social isolation was associated with a highly significant elevation in the proportion of striatal D(2) (High), equivalent to a 3.3-fold increase (group 15.2% +/- 1.4%, isolate 49.8% +/- 4.8%; P < 0.0001, Student's unpaired t-test). These findings support both the hypothesis that elevated D(2) (High) is a common feature of multiple animal models of psychosis, and the validity of isolation rearing as a neurodevelopmental model of a "schizophrenia-like" state. PMID- 19217029 TI - Value of cytopathologist-performed ultrasound-guided fine-needle aspiration as a screening test for ultrasound-guided core-needle biopsy in nonpalpable breast masses. AB - Fine-needle aspiration (FNA) of breast masses in the United States has been on the decline for the last decade and has been largely replaced by ultrasound guided core-needle biopsy (UG-CNB). Some studies show core-needle biopsy (CNB) is superior to FNA in terms of absolute sensitivity, specificity, and inadequate rate. However, the importance of a skilled aspirator, experienced cytopathologist, and immediate cytological evaluation (ICE) in FNA is often not considered. CNB is more expensive, invasive, risky, and painful than FNA. This prospective study examines the value of cytopathologist-performed ultrasound guided FNA (UG-FNA) with ICE as a screening test for cytopathologist-performed UG CNB on nonpalpable or difficult-to-palpate solid breast masses visible on ultrasound. One hundred twenty consecutive nonpalpable or difficult-to-palpate presumably solid breast masses in 109 female patients from January2, 2008 to June 30, 2008 underwent cytopathologist-performed UG-FNA with ICE. Twenty cases were converted to cytopathologist-performed UG-CNB because ICE was inadequate, hypocellular, atypical, suspicious, or malignant. Patients with clearly benign cytology did not undergo UG-CNB. UG-FNA with ICE reduced the percentage of patients undergoing UG-CNB by 87%. A new role for cytopathologist-performed UG FNA of nonpalpable breast masses has been identified. PMID- 19217030 TI - Immunocytochemical panel for distinguishing between adenocarcinomas and reactive mesothelial cells in effusion cell blocks. AB - The aim of our study was to determine the value of a panel that consisted of one epithelial marker (MOC-31) and two mesothelial markers (D2-40 and calretinin) for distinguishing between reactive mesothelial cells (RMCs) and adenocarcinomas (ACs) in effusion fluids. A total of 118 cell block specimens from pleural and peritoneal effusions, including 88 ACs and 30 benign effusions with RMCs were stained with antibodies against MOC-31, D2-40, and calretinin. MOC-31 membranous activity was observed in all samples from ACs, regardless of the primary tumor site. All benign effusion samples with RMCs were negative for MOC-31. All benign effusion samples with RMCs exhibited membranous staining for D2-40, and one AC case had focal reactivity for D2-40. Almost all benign effusions reacted positively with calretinin. Staining was noted in both the cytoplasm and the nucleus in the majority of cases. Scattered tumor cells had weak calretinin positivity in two AC cases. Background RMCs in AC effusions were consistently positive for D2-40 and calretinin. In general, D2-40 identified more RMCs than calretinin. The staining combination of positive for MOC-31 and negative for D2 40 or calretinin were 100% specific and 99% sensitive for ACs. Our data suggest that immunohistochemical studies performed on cell blocks with MOC-31, D2-40, and calretinin were useful in the differentiation between ACs and RMCs. D2-40 was a more sensitive marker for RMCs than calretinin. PMID- 19217031 TI - Evaluation of new monoclonal antibodies in detection of estrogen receptor, progesterone receptor, and Her2 protein expression in breast carcinoma cell block sections using conventional microscopy and quantitative image analysis. AB - Accurate assessment of estrogen receptor (ER), progesterone receptor (PR), and Her2 status of breast carcinomas is critical for predicting response to systemic therapies. Recently, developed rabbit monoclonal antibodies (RMab) are reported to have higher sensitivity than murine monoclonal antibodies (Mab). This study compares RMabs against FDA-approved Mab (FMab) in breast carcinoma cell block sections using visual and image quantification. Cell blocks from 52 breast cancers were studied. Immunohistochemistry using RMab (ER, PR, and Her2) was compared with FMabs (ER, PR, Dako) and HercepTest (HerFDA). Fluorescent in situ hybridization (FISH) was used as a reference standard for Her2. Slides were later scanned and reanalyzed with an automated cellular imaging system (ACIS III, Dako). Frequency of ER (38.5% vs. 36.5% for visual; 55.8% vs. 57.7% for image) and PR (28.8% vs. 36.5% for visual; 50% vs. 51.9% for image), and concordance (overall agreement is 71.2% and 75% for visual and image ER; and 84.6% and 59.6% for visual and image PR) were similar for both FMab and RMab, respectively. Overall agreement (53.8% vs. 77.1% for visual and image detection, respectively, using HerFDA and RMab) is poor to moderate for Her2. Visual Her2 (RMab) has the highest concordance (94.1%), and visual HerFDA has the lowest concordance (35.3%) with FISH. ER and PR analysis (FMab vs. RMab) are almost comparable using both detection methods with good overall agreement. For Her2 overexpression, RMab proved to be superior to HerFDA and showed excellent agreement with FISH results with both quantitative detection methods. PMID- 19217032 TI - Papillary thyroid carcinoma with atypical histiocytoid cells on fine-needle aspiration. AB - Although papillary thyroid carcinoma (PTC) usually has classic cytological characteristics on fine-needle aspiration (FNA), it can present rarely with aberrant features resembling those of histiocytes in a cystic nodule. The aim of the current study was to describe PTC with atypical histiocytoid cells and distinguish it from benign histiocytes. A retrospective computerized search for FNAs with atypical features suggestive of PTC and cystic degeneration was performed, and if available, the corresponding resection specimens were compared. Four cases met the criteria for FNAs and three had surgical pathology follow-up, which showed PTC. One aspirate had some features typical of PTC, but the remaining FNAs had atypical histiocytoid cells, which had traits intermediate between those of PTC and histiocytes. Large cell size, pseudoinclusions, nuclear grooves, and multiple well-defined vacuoles in atypical histiocytoid cells favor PTC over benign histiocytes. Ancillary immunocytochemical studies can also be useful in confirming the diagnosis. Histiocytic cells are frequently present in thyroid aspirates, and occasionally, they have atypical features that represent an unusual presentation of PTC. Closer examination of these cells can provide diagnostic clues for preventing false-negative diagnosis of PTC. PMID- 19217033 TI - Has the ThinPrep method of cervical screening maintained its improvement over conventional smears in terms of specimen adequacy? AB - Liquid-based cytology (LBC) has replaced conventional smear assessment in many centers over recent years. In our laboratory this transfer took place in 1999. At that time we performed a split sample study comparing the conventional method of cervical smear evaluation with the ThinPrep system. This split sample study identified a dramatic improvement in specimen adequacy with LBC. While 11% of conventional preparations were reported as unsatisfactory and almost 9% were reported as suboptimal, evaluation of the same cases using LBC saw this combined figure reduced to 2.3%. AIM: To evaluate whether this dramatic fall in unsatisfactory smears has been maintained with the use of LBC. The database for all smears reported for 2005 (100% LBC) was interrogated. The number of unsatisfactory reports was calculated. The reason for an unsatisfactory report was recorded for each case. The overall unsatisfactory rate was compared with that reported in the 1999 split sample study. A total of 41,312 smear tests were reported in 2005. 1,342 (3.25%) were reported as unsatisfactory. Our findings support the ongoing value of LBC in a routine cervical screening laboratory in terms of continuing to maintain a low rate of unsatisfactory smears. PMID- 19217034 TI - Comparison of accuracy of diagnostic modalities for evaluation of breast cancer with review of literature. AB - This study was conducted to confirm which of these modalities [mammography (MG), fine-needle aspiration cytology (FNAC), core-needle biopsy (CNB), or intraoperative touch imprint cytology (IOTIC)] is useful to detect breast cancer and to compare the accuracy of these modalities for the diagnostic setting. One hundred seventy-five, 85, 78, and 25 patients were selected who underwent FNAC, MG, IOTIC, and CNB, respectively. Histopathology was used as a gold standard for comparison of the results of all diagnostic modalities. Twenty-five patients with combinations of three preoperative tests (MG, FNAC, CNB) showed 100% accurate results when compared with histopathology, while the overall accuracy for MG, FNAC, and CNB was 91.7%, 91.5%, and 96%, respectively. Both touch and scrape imprints' sensitivity, specificity, positive predictive value, negative predictive value (NPV), and accuracy were 100% for class V and class II smears, while sensitivity, accuracy, and NPV were 87, 95, and 83% for IOTIC and 94%, 96%, and 89% for IOSC for class III and IV smears. On comparison of the results of all these procedures, the difference was nonspecific (P = 0.2, 0.3, 0.5, 0.55, and 0.6 for MG, FNAC, IOTI, IOSC, and CNB, respectively). Combinations of preoperative tests (MG, FNAC, CNB) were more accurate, reliable, and acceptable when compared with individual diagnostic procedure, but these have their own technical limitations. The accuracy of CNB was much higher than that of MG and FNAC. IOTP and IOSC are simple and cost effective diagnostic tests with better accuracy that can be used as an alternate to frozen section in diagnostic settings and margin assessments. PMID- 19217035 TI - Ascitic fluid cytology of a malignant mixed Mullerian tumor of the peritoneum: a report of two cases with special reference to p53 status. AB - Malignant mixed mullerian tumors (MMMTs) rarely originate in the female peritoneum. Peritoneal MMMT, as well as its uterine or ovarian counterpart, is characterized by biphasic histologic components, including carcinoma of the mullerian type and sarcoma. In ascitic fluid cytology of MMMT, however, the biphasic pattern is less evident than in tissue sections, and heterogeneity of cell differentiation makes the cytologic diagnosis difficult, especially in distinguishing tumor cells from reactive mesothelial cells. Here, we report ascitic fluid cytology for two peritoneal MMMT cases. Immunocytochemistry for p53 was helpful in identifying malignant cells; tumor cells in ascitic fluid smears, as well as carcinomatous cells and sarcomatous cells in tissue sections, showed distinct nuclear immunostaining for p53, whereas mesothelial cells did not. Subsequent molecular genetic analysis confirmed frameshift mutations in both cases. To the best of our knowledge, this is the first report showing p53 overexpression and its genetic background in MMMT of the peritoneum. PMID- 19217036 TI - Cytologic diagnostic pitfall of dermatofibrosarcoma protuberans masquerading as primary parotid tumor: a case report. AB - We report the cytology of a rare case of dermatofibrosarcoma protuberans (DFSP) involving the parotid region. Our patient was a 55-year-old female who presented with a swelling in the right parotid region, which was clinically interpreted as a "recurrent parotid tumor." Fine needle aspiration revealed a moderate cellular yield with spindle cell component, seen discretely as well as, embedded loosely within a fibrillary magenta matrix. A few cellular spindle cell fascicles and several discrete, naked, spindly nuclei were also noted. Neoplastic cells exhibited a minimal pleomorphism with a relatively bland chromatin and inconspicuous nucleoli. A differential diagnosis of the benign spindle cell tumor/low, or intermediate grade myxoid, spindle cell sarcomas was considered. Owing to its clinical presentation simulating a parotid tumor, possibility of "spindle cell myoepithelioma" was suggested. However, histopathologic examination with immunohistochemistry showed it to be a DFSP. A precise diagnosis of DFSP is not always possible on cytology alone. Owing to its location, as well as the spindle cell morphology, DFSP occurring in the parotid region is likely to be mistaken for a spindle cell myoepithelioma, in which case, an appropriate panel of immunomarkers is required for resolving the diagnostic dilemma. PMID- 19217037 TI - Cytomorphologic features of the plasmacytoid variant of urothelial carcinoma: a case report. AB - The plasmacytoid variant of urothelial carcinoma is an uncommon form of urothelial carcinoma with a distinctive histologic appearance. Descriptions of the cytologic features of this particular type of urothelial carcinoma in urinary specimens are limited. The present report details the urinary cytomorphologic findings of a plasmacytoid urothelial carcinoma occurring in the urinary bladder of a 66-year-old male. Bladder washings showed the presence of scattered dyscohesive cells with a plasmacytoid appearance characterized by abundant amounts of cytoplasm, eccentrically located nuclei, and inconspicuous nucleoli. The diagnosis of plasmacytoid urothelial carcinoma was confirmed upon histologic examination of a concurrently performed transurethral resection specimen, with immunohistochemical studies supporting this interpretation. Although rare, the plasmacytoid variant of urothelial carcinoma should be a diagnostic consideration when atypical cells with plasmacytoid features are encountered in a urinary specimen. PMID- 19217038 TI - Aspergillus/Penicillium sp. spores as a contaminant on conventional Pap smear. PMID- 19217039 TI - A case of urothelial carcinoma with triple variants featuring nested, plasmacytoid, and lipoid cell morphology. AB - We present very rare variants of urothelial carcinoma featuring nested, plasmacytoid, and lipoid cell morphology in an 80-year-old female who was admitted to our hospital with mictritional pain and bilateral hydronephrosis. Abdominal computed tomography showed diffuse thickening of the urinary bladder, indicating probable invasion into the surrounding adipose tissue. Cytological tests on a urine specimen revealed medium-sized cancer cell clusters with hyperchromatic nuclei and dense cytoplasm, small clusters of dyshesive cells with eccentric or crescent nuclei, and abundant or vacuolated cytoplasm mimicking plasma cells or lipoblasts. Histopathological findings of the bladder tumor revealed that the nested urothelial carcinoma variants were mainly located in the lamina propria, whereas the plasmacytoid and lipoid-cell variants had deeply infiltrated into cells with a myxoid background. Immunohistochemically, the cancer cells were positive for cytokeratin, epithelial membrane antigen, but not for vimentin, S-100, or hematopoietic markers of plasma cells such as CD79alpha and CD38. The patient received no radiosurgical therapies because of the advanced stage of her disease and died a few months after the initial diagnosis. To our knowledge, this is the first cytological and histological examination of urothelial carcinoma consisting of nested, plasmacytoid, and lipoid-cell variants of the urinary bladder. PMID- 19217040 TI - Dense deposits in cervical cytology samples of patients fitted with intrauterine contraceptive devices. PMID- 19217041 TI - Cytologic and immunocytochemical features of EBV negative primary effusion lymphoma: report on seven Japanese cases. AB - Primary effusion lymphoma (PEL) is very rare type of non-Hodgkin's lymphoma (NHL) usually confined to the body cavities such as the pleural space, pericardium, and peritoneum. PEL is a human herpes virus-8 (HHV-8)-associated lymphoma and commonly observed in human immunodeficiency virus (HIV)-infected patients. However, HIV-infected patients are extremely fewer in Japan in comparison with those in Western countries; PEL is usually not associated with HIV infection in Japan. This report presents seven Japanese cases of PEL. In situ hybridization revealed that the PEL cells were negative for EBV in all cases. An immunocytological analysis showed that only one case was positive for HHV-8, and PEL cells were positive for CD20 in all cases. MUM1 was positive, but CD10 and CD138 were negative in six cases. One case each was positive for CD30 and BCL-6. The phenotypic patterns of HIV-related is BCL6-/MUM1+/CD138+, thus, the phenotypic findings observed by immunocytochemistry in this study were somehow different from those reported in Western countries. However, the cytomorphological features of PEL cells showed large cell size, abundant basophilic cytoplasm, coarse chromatin, and occasional binucleated or multinucleated cells, similar to a large cell immunoblastic and anaplastic large cell lymphoma, indicating that the cytomorphological characteristics of PE cells in Giemsa and Papanicolaou stain were consistent with those reported abroad. The prognosis for PEL in these cases was poor, but the survival time was variable ranging from 1 month to 54 months, and was different from that of Western cases. No p16/CDKN2A expression was observed, and one case showed PEL cells with a BLIMP1 mutation. PMID- 19217042 TI - Nodular pulmonary amyloidosis with an unusual protein composition diagnosed by fine-needle aspiration biopsy: a case report. AB - Nodular pulmonary amyloidosis is rare, but a number of individual cases have been reported in the literature. Radiologically, pulmonary amyloid nodules often mimic cancer and hence require biopsy to obtain a specimen for definitive diagnosis. We describe a case of isolated nodular pulmonary amyloidosis with an unusual amyloid composition that given the patient's clinical history has not been described in the literature to the best of our knowledge, making this case unique. A 58-year old woman with a history of mixed connective tissue disease was found to have multiple lung nodules on chest computed tomography (CT) that were radiologically suspicious for cancer. A CT-guided percutaneous fine-needle aspiration (FNA) biopsy of one of the nodules was performed. A diagnosis of amyloid was rendered using special stains and polarized light microscopic examination. Immunohistochemical stains revealed that the amyloid comprised beta-2 microglobulin as well as both kappa and lambda light chains. Further work up showed no evidence of systemic amyloidosis. We were successful in rendering an accurate diagnosis of a pulmonary amyloidoma on material procured by CT-guided FNA thus avoiding more invasive procedures. In addition, immunohistochemical staining revealed an unexpected and highly unusual protein composition of the amyloid. PMID- 19217043 TI - Psammoma bodies in cervical smear in association with keratinizing squamous cell carcinoma of cervix: a case report. AB - The presence of psammoma bodies (PBs) in cervical smears is a rare finding. These structures have been identified in association with a wide range of benign and malignant conditions within the female genital tract. PBs in cervical smears have usually been associated with malignant serous epithelial ovarian tumors. However, many PBs associated with atypical squamous cells were detected in cervical smears of an 83-year-old woman with complaint of postmenopausal bleeding. Colposcopic examination revealed an ulceroinfiltrative growth in the cervix. Histological examination of the biopsy specimen from the growth revealed keratinizing squamous cell carcinoma with multiple and singly arranged PBs. This report suggests that cytologists should aware of the possibilities, on finding PBs associated with atypical cells in cervical specimens and report the cases accordingly. PMID- 19217044 TI - Hymenolepis nana in squash preparation at endoscopy. PMID- 19217045 TI - Cytology of metastatic cervical squamous cell carcinoma in pleural fluid: report of a case confirmed by human papillomavirus typing. AB - Cervical squamous cell carcinomas are rarely the cause of malignant effusions. Their identification can be relatively easy when keratinizing atypical squamous cells are present, but may be very difficult when only nonkeratinizing malignant cells are present. We present the case of a 47-year-old woman who presented with a large left pleural effusion after having recently completed chemoradiation therapy for stage IIB cervical squamous cell carcinoma. Cytologic examination of the fluid showed a uniform population of single atypical cells with finely vacuolated cytoplasm, ectoendoplasmic demarcation, cell-in-cell arrangements, and short rows of cells with intervening "windows," all features reminiscent of mesothelial cells. No keratinization or three-dimensional cell clusters were identified. A panel of immunohistochemical stains was performed on the cell block material, and the atypical cells were positive for cytokeratin 5/6, p63, and p16 but not for cytokeratin 7, calretinin, WT1, or Ber-EP4 or TTF1. These findings were consistent with metastatic squamous cell carcinoma. HPV DNA determination and typing by PCR confirmed the presence of HPV16 in an aliquot of pleural fluid. This is to our knowledge the first reported case of pleural fluid involved by metastatic squamous cell carcinoma where HPV DNA testing was used to confirm the origin of the metastasis. Despite its rarity, metastatic nonkeratinizing squamous cell carcinoma should be considered when a single cell population of large atypical cells is found in effusions. Immunoperoxidase stains and HPV testing can be performed to establish the diagnosis and confirm the origin from a cervical primary. PMID- 19217046 TI - Cellular mesoblastic nephroma in an infant: report of the cytologic diagnosis of a rare paediatric renal tumor. AB - Congenital mesoblastic nephroma is a rare pediatric tumor with a favorable clinical outcome. Cytological features of this uncommon tumor and diagnostic difficulties with other commoner pediatric renal neoplasms have been inadequately discussed in the available literature. We describe the case of a 1-year-old girl who presented with a right renal mass. Fine-needle aspiration smears consisted of a few cellular clusters of spindle cells with mitotic activity and mild nuclear pleomorphism. No blastema was identified. A cytologic impression of mesoblastic nephroma was rendered, which was confirmed on histopathological examination of the right nephrectomy specimen as a cellular mesoblastic nephroma. Cytologic diagnosis of mesoblastic nephroma has important prognostic and therapeutic implications. The cytopathologist should carefully evaluate smears from such patients and attempt to differentiate mesoblastic nephroma from Wilms' tumor and clear cell sarcoma. PMID- 19217047 TI - Decidualized endometrioma diagnosed by fine needle aspiration cytology: a case report with immunocytochemical confirmation. AB - Endometriosis is defined as ectopic endometrial tissue which can respond to hormonal stimulation. Cutaneous endometriosis is a rare pathologic entity that can represent a clinical diagnostic challenge. We report a case of a decidualized endometrioma in a 24-year old pregnant African American woman diagnosed by fine needle aspiration cytology and confirmed by immunocytochemistry using CD10, ER, and Calretinin. The awareness of the pathologist of the cytologic characteristics of this uncommon entity is important to avoid diagnostic pitfalls and unnecessary diagnostic interventions during pregnancy. PMID- 19217048 TI - Ulcerated neck mass and testicular carcinoma. PMID- 19217049 TI - Endoscopic ultrasound-guided fine-needle aspiration diagnosis of clear-cell pancreatic endocrine neoplasm in a patient with von Hippel-Lindau disease: a case report. AB - The cytologic findings of a clear-cell pancreatic endocrine neoplasm (PEN) diagnosed by endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) occurring in a 29-year-old man with von Hippel-Lindau (VHL) syndrome are described. Imaging studies showed multiple pancreatic masses and cysts, a single liver lesion, and pulmonary and renal cysts. Cytologic features of this clear cell PEN included the presence of large sheets and rounded clusters of polygonal neoplastic cells with relatively abundant cytoplasm containing numerous, small, sharply-demarcated vacuoles that occasionally indented nuclei and gave the cells a "frothy" appearance. Mild anisonucleosis was present and nucleoli were visible. Rare single cells and stripped nuclei were seen. Small vessels transgressed tumor cell sheets. These cytologic findings are distinct from those of typical endocrine neoplasms, and bear resemblance to the cytologic features of renal cell carcinoma metastatic to the pancreas. To the best of our knowledge, this is the first detailed report of the EUS-FNA cytologic findings of a clear-cell PEN associated with VHL syndrome. We believe that the distinctive and characteristic cytologic features, together with immunohistochemical studies, can allow a preoperative cytologic diagnosis of this highly unusual pancreatic lesion and avoid possible confusion with other clear-cell neoplasms in the pancreas, particularly metastatic renal cell carcinoma. PMID- 19217050 TI - Pitfalls in the diagnosis of adult rhabdomyoma by fine needle aspiration: report of a case and a brief literature review. AB - Fine needle aspiration (FNA) is a valuable, noninvasive, commonly used technique in the diagnoses of head and neck tumors. Adult rhabdomyoma is a rare benign tumor of striate muscle tissue usually located in the head and neck region. Cytomorphologically, the tumor cells have eosinophilic finely granular cytoplasm, which sometimes can mimic other tumors. We report a case of adult rhabdomyoma, which was initially misinterpreted as granular cell tumor on FNA biopsy due to lacking of typical cytological features of adult rhabdomyoma. We recommend that adult rhabdomyoma should be included in differential diagnoses when the cytological features are suggestive of granular cell tumor. A panel of immunohistochemical stains including S100, desmin, and myoglobulin may also be helpful in making correct diagnosis. PMID- 19217051 TI - "Fruiting body" Penicillium species: Papanicolaou stained conventional cervical smear findings. PMID- 19217052 TI - Pure neuritic leprosy with nerve abscess presenting as a cystic, soft tissue mass: report of a case diagnosed by fine needle aspiration cytology. AB - Pure neuritic leprosy (PNL) with nerve abscess manifesting as a huge, cystic, soft tissue mass is highly uncommon. Fine needle aspiration cytology can serve as an important initial diagnostic modality in such an instance. We report a case of 28-year-old male, who presented with a huge swelling in the lower, medial aspect of the right upper arm. The clinical diagnosis was schwannoma. Fine needle aspiration (FNA) yielded 80 ml of sticky, turbid, pale brown fluid. Cytologic examination revealed abundant, caseous, necrotic material and many degenerated neutrophils in a thin proteinaceous background. Stain for acid fast bacilli (AFB) was negative. Based on an AFB negative, caseous, necrotic material obtained from the soft tissue mass located in the ulnar nerve region, a cytodiagnosis of tuberculoid PNL with nerve abscess was given, and this was confirmed by the subsequent histopathologic examination. Our case emphasizes the major role of minimally traumatic, FNA technique in the diagnosis of rare cases of clinically unsuspected neuritic leprosies. PMID- 19217053 TI - Mucoepidermoid thymic carcinoma: a challenging mediastinal aspirate. AB - Primary thymic carcinoma-mucoepidermoid cell (MEC) type is rare and only one report describing the cytologic features of this neoplasm in the metastatic site is described. We describe the cytological features of poorly differentiated carcinoma possibly MEC in a 54-year-old man who presented with cough, weight loss, and puffiness of face for 3 months. The significance of this infrequently encountered neoplasm lies in its potential confusion of origin of the tumor thymus or metastases from a primary bronchial MEC. Immunocytochemical profile was suggestive of a thymic carcinoma of the MEC type. PMID- 19217054 TI - Imprint smear cytology of chondroblastic osteosarcoma: a case report. AB - Imprint cytology is gaining increasing popularity in the diagnosis of musculoskeletal lesions and in many patients; a definitive diagnosis can be rendered from aspiration smears alone. However, evaluation of tissue architecture is a major limitation and therefore cytological diagnosis has to be confirmed on histopathology. The authors take this opportunity to report a case of chondroblastic osteosarcoma occurring in the middle 1/3rd of the left thigh in a 35 year old male, diagnosed on imprint cytology and confirmed on histopatholgy. PMID- 19217056 TI - Cytodiagnosis of actinomycetoma. AB - Mycetoma is a late clinical manifestation of a subcutaneous infection produced by bacteria (actinomycetoma) or fungi (eumycetoma). The distinction between eumycetoma and actinomycetoma in Fine Needle Aspiration Cytology (FNAC) is as accurate as histopathology. A 55 year old man presented with a slow growing swelling on the plantar aspect of the right foot which was present for the last 10 years. A clinical diagnosis of soft tissue tumor was made and FNAC was advised. Smears revealed mixed inflammatory infiltrate and foreign body type of giant cells along with clumps of fibrillar organisms. Gram stain done later demonstrated gram positive thin branching filaments. A diagnosis of actinomycetoma was rendered. Histopathology of the excised specimen confirmed the cytologic diagnosis of actinomycetoma. Mycetoma can be accurately diagnosed by FNAC, which is a simple, inexpensive technique for rapid diagnosis. PMID- 19217055 TI - Diagnostic utility of mammaglobin and GCDFP-15 in the identification of metastatic breast carcinoma in fluid specimens. AB - Morphologic differentiation of breast carcinoma from nonmammary malignancies in fluid specimens can be a diagnostic challenge. Immunocytochemistry is often employed in the differential diagnosis. In this study, we evaluated the expression of mammoglobin (MGB1) in body-cavity fluid specimens and compared its efficacy as a marker for metastatic breast carcinomas with that of gross cystic disease fluid protein-15 (GCDFP-15). Cell blocks from 40 fluid specimens were immunostained with monoclonal antibodies against MGB1 and GCDFP-15. They included 15 breast carcinomas and 25 nonmammary carcinomas (10 lungs, 10 ovaries, 3 gastrointestinal tracts, 1 kidney, and 1 urinary bladder). Positivity was defined as the presence of cytoplasmic staining in 10% or more carcinoma cells. Thirteen (87%) and seven (47%) breast carcinomas showed positive staining with MGB1 and GCDFP-15, respectively. Three (12%) nonmammary carcinomas (2 ovarian and 1 colonic) showed positive MGB1 staining; one (3%) nonmammary carcinoma demonstrated positive GCDFP-15 staining. The differences of MGB1 and GCDFP-15 staining between breast and nonmammary carcinomas were statistically significant (P < 0.05). Both MGB1 and GCDFP-15 are specific markers for metastatic breast carcinomas in cell block fluid specimens (88 vs. 96%). However, MGB1 is more sensitive than GCDFP-15 as a marker for metastatic breast carcinoma (87 vs. 46%). PMID- 19217057 TI - Comparison of EUS-guided FNA and Trucut biopsy for diagnosing and staging abdominal and mediastinal neoplasms. AB - The objective of this study was to evaluate endoscopic ultrasound Trucut biopsy (TCB) specimens and compare these findings to fine needle aspiration (FNA) specimens for the diagnosis of neoplasia. FNA and TCB specimens were reviewed in blinded fashion by a cytopathologist from patients (N = 93) who had EUS-guided FNA and TCB specimens collected between July 2000 and January 2005. Specimens were categorized as nondiagnostic, negative, suspicious for stromal neoplasm, suspicious for malignancy, positive for stromal neoplasm, or positive for malignancy. Standard final diagnosis based on clinical and/or pathologic follow up was available for 86 of 93 patients. The final diagnoses comprised malignancy (n = 55), stromal neoplasm (n = 19), and benign findings (n = 12). The combination of FNA and TCB results combined were significantly (P < 0.001) more sensitive that FNA alone for the detection of both malignancy (78% vs. 55%) and stromal neoplasia (79% vs. 19%) without a significant change in overall specificity (92% vs. 100%, P = 1.00). A positive FNA specimen with a negative/nondiagnostic TCB result was established in seven patients with malignancy. A positive TCB diagnosis with a negative/nondiagnostic FNA result was noted in five patients with malignancy. A suspicious FNA result was upgraded to positive in conjunction with TCB specimen evaluation in eight patients with malignancy. The results of this study suggest that TCB is a useful adjunctive technique when used in tandem with FNA for malignancy and stromal neoplasia detection. Additional data are needed to firmly establish practice guidelines for the use of EUS-guided TCB specimens in clinical practice. PMID- 19217058 TI - Comparison of cytological results obtained by repeated US-guided fine-needle aspiration biopsies of thyroid nodules: focus on the rate of malignancy and diagnostic concordance. AB - The debate still continues on the repeated fine-needle aspiration biopsies (FNABs) for thyroid nodule in clinical practice. In this study, we determined the rate of cytological change to malignancy and the diagnostic concordance of repeated FNABs when the same nodules were targeted under US-guidance. We retrospectively reviewed data for 187 thyroid nodules (173: twice, 14: three times) from 160 patients who underwent repeated US-guided FNABs, which were performed by one skillful radiologist targeting for the same nodules at a mean interval of 7.5 months. Their initial cytological findings were compared with second or third results and histopathologic follow-up. The initial FNABs findings of 187 nodules were unsatisfactory, benign, and indeterminate in 56, 52, and 79 cases, respectively. The rate of a second cytological diagnosis changed to malignancy was significantly higher in the unsatisfactory aspirates (10.7%; 6/56), when compared with those of the benign (0.0%; 0/52) or of indeterminate aspirates (3.8%; 3/79) (P = 0.022). However, there was no change to malignancy at third cytological findings of all 14 nodules. After the second US-guided FNABs, 30.8% (16/52) of the initially diagnosed as benign aspirates were reclassified as indeterminate, while 26.6% (21/79) of the initially diagnosed as indeterminate were reclassified as benign. In conclusion, to identify malignancies, repeated US guided FNABs are recommended for thyroid nodules initially classified as unsatisfactory aspirates. However, although US-guidance is applied, a discrepancy might be unavoidable in the cytological interpretation of the nodules classified as benign or as indeterminate aspirates because of overlapping cytological criteria. PMID- 19217059 TI - Breast Actinomyces neuii abscess simulating primary malignancy: a case diagnosed by fine-needle aspiration. PMID- 19217060 TI - Evaluation of flow cytometric immunophenotyping and DNA analysis for detection of malignant cells in serosal cavity fluids. AB - The serosal cavities are frequent sites of tumor metastasis. The distinction between carcinoma cells, inflammatory cells, and reactive or malignant mesothelial cells can be difficult in cytology. Multicolor flow cytometry (FCM) provides the opportunity to evaluate multiple antigens simultaneously, making it possible to characterize various cell populations. In this study, we aimed to assess the diagnostic accuracy of FCM immunophenotyping and DNA in comparison with serum tumor markers and classic cytology for detection of malignant cells in pleural and ascitic fluids. One hundred and nineteen samples of body cavity fluids were analyzed. Immunophenotyping was performed by four-color immunofluorescent staining using monoclonal antibodies against Ber-EP4, cytokeratin, CD3, and CD45. The DNA analysis by FCM was also performed. In addition, serum CA19-9, CEA, AFP, and CA125 were analyzed. Ber-EP4 marker had the highest sensitivity (73%) and specificity (95.5%) in the detection of carcinoma cells in serous fluid and correlated with cytology in most of cases (73%). The mean of DI differed statistically in patients with malignant effusions than in benign one. DI showed no difference in fluids due to infiltration of malignant epithelial cells or hematopoietic malignancy or due to hepatocellular carcinoma developing in cirrhotic liver. Thus, flow cytometry appears to aid not only in the detection of malignant cells but also in the characterization of cell type. On the other hand, although DNA ploidy examination had better sensitivity; it had no advantage over conventional cytopathological examination in identification of malignant cells. PMID- 19217061 TI - Oncocytoma of the lacrimal gland diagnosed initially by fine-needle aspiration cytology. AB - Oncocytomas are benign tumors that infrequently involve ocular adnexa. We describe a case of oncocytoma of the lacrimal gland which was initially diagnosed by fine-needle aspiration cytology. An 83-year-old patient presented to the cytology laboratory with a peripunctal mass in the right eye. The tumor was aspirated. The cytoplasm of the tumor cells was rich in eosinophilic granules. The initial cytological diagnosis was oncocytoma. The tumor was then completely excised and the histological diagnosis confirmed the initial cytological one. Oncocytoma is a rare entity which must be considered in differential diagnosis of a peripunctal mass in elderly patients. The literature was reviewed confirming the rarity of such a presentation and the novelty of this case. Cytological criteria of malignancy are not yet established. PMID- 19217062 TI - Low value of color flow-doppler in predicting malignancy of thyroid follicular neoplasms. PMID- 19217063 TI - Fine-needle aspiration biopsy diagnosis of prostate carcinoma metastatic to the breast. PMID- 19217064 TI - An approach to post-radical trachelectomy vaginal-isthmus cytology. AB - Radical trachelectomy (RT) is the surgical amputation of the uterine cervix with paracervical lymphadenectomy, performed in reproductive age women to treat invasive squamous-cell carcinoma or endocervical adenocarcinoma while preserving the uterine corpus for potential child bearing. Post-RT patient monitoring includes isthmic-vaginal cytology. This study reviews our experience with liquid based preparation of post-RT cytology samples. Fifty-four post-RT vaginal-isthmic cytology specimens were reviewed from nine patients, seven with adenocarcinoma, and two with squamous-cell carcinoma. Five patients had normal (NILM) or normal with reactive changes on all cytology samples. Two patients had isolated squamous abnormalities (atypical squamous-cells of uncertain significance (ASC-US) and low grade squamous intraepithelial lesion (LSIL)); both follow-up biopsies were negative. Two patients had repeatedly abnormal specimens interpreted as atypical glandular cells (AGC), one of whom also had a concurrent ASC-US. Only one sample was tested for high risk human papilloma virus (hrHPV), with negative results. All patients with abnormal cytology went on to have biopsies which were interpreted as benign. The cytology specimens most often interpreted as AGC contained many groups of hyperchromatic crowded glandular cells and/or stromal cells derived from direct sampling of the lower uterine segment. The crowding often limits visualization of all the cells in a group, plus sampled endometrium may harbor mitoses, adding to the atypical appearance. Cytologists should become familiar with the spectrum of changes in the post-RT cytology. Testing for hrHPV should be considered for use in the management of abnormal cytology results. Post RT cytology should be compared with presurgical cytology since one would anticipate similarities in post-RT true positive cases. In particular, a primary diagnosis of adenocarcinoma makes differentiating benign reactive glandular cells from recurrence a critical issue. PMID- 19217065 TI - Anatomical Sciences Education: a solid contribution during its first year. PMID- 19217066 TI - Evaluation of computer-aided instruction in a gross anatomy course: a six-year study. AB - Web-based computer-aided instruction (CAI) has become increasingly important to medical curricula. This multi-year study investigated the effectiveness of CAI and the factors affecting level of individual use. Three CAI were tested that differed in specificity of applicability to the curriculum and in the level of student interaction with the CAI. Student personality preferences and learning styles were measured using the Meyers Briggs Type Indicator (MBTI) and Kolb's Learning Style Inventory (LSI). Information on "computer literacy" and use of CAI was collected from student surveys. Server logs were used to quantify individual use of respective CAI. There was considerable variability in the level of utilization of each CAI by individual students. Individual use of each CAI differed and was associated with gender, MBTI preferences and learning style, but not with "computer literacy." The majority of students found the CAI useful for learning and used the CAI by themselves. Students who accessed the CAI resources most frequently scored significantly higher on exams compared with students who never accessed the resources. Our results show that medical students do not uniformly use CAI developed for their curriculum and this variability is associated with various attributes of individual students. Our data also provide evidence of the importance of understanding student preferences and learning styles when implementing CAI into the curriculum. PMID- 19217067 TI - A new presentation and exploration of human cerebral vasculature correlated with surface and sectional neuroanatomy. AB - The increasing complexity of human body models enabled by advances in diagnostic imaging, computing, and growing knowledge calls for the development of a new generation of systems for intelligent exploration of these models. Here, we introduce a novel paradigm for the exploration of digital body models illustrating cerebral vasculature. It enables dynamic scene compositing, real time interaction combined with animation, correlation of 3D models with sectional images, quantification as well as 3D manipulation-independent labeling and knowledge-related meta labeling (with name, diameter, description, variants, and references). This novel exploration is incorporated into a 3D atlas of cerebral vasculature with arteries and veins along with the surrounding surface and sectional neuroanatomy derived from 3.0 Tesla scans. This exploration paradigm is useful in medical education, training, research, and clinical applications. It enables development of new generation systems for rapid and intelligent exploration of complicated digital body models in real time with dynamic scene compositing from highly parcellated 3D models, continuous navigation, and manipulation-independent labeling with multiple features. PMID- 19217068 TI - Design and development of a new facility for teaching and research in clinical anatomy. AB - This article discusses factors in the design, commissioning, project management, and intellectual property protection of developments within a new clinical anatomy facility in the United Kingdom. The project was aimed at creating cost effective facilities that would address widespread concerns over anatomy teaching, and support other activities central to the university mission-namely research and community interaction. The new facilities comprise an engaging learning environment and were designed to support a range of pedagogies appropriate to the needs of healthcare professionals at different stages of their careers. Specific innovations include integrated workstations each comprising of a dissection table, with removable top sections, an overhead operating light, and ceiling-mounted camera. The tables incorporate waterproof touch-screen monitors to display images from the camera, an endoscope or a database of images, videos, and tutorials. The screens work independently so that instructors can run different teaching sessions simultaneously and students can progress at different speeds to suit themselves. Further, database access is provided from within an integrated anatomy and pathology museum and display units dedicated to the correlation of cross-sectional anatomy with medical imaging. A new functional neuroanatomy modeling system, called the BrainTower, has been developed to aid integration of anatomy with physiology and clinical neurology. Many aspects of the new facility are reproduced within a Mobile Teaching Unit, which can be driven to hospitals, colleges, and schools to provide appropriate work-based education and community interaction. PMID- 19217069 TI - Privileged learning. AB - Students often attain memorable experiences from cadaver dissections through reflective writing. For many, facing a dissection for the first time elicits a wide range of emotions. These may include thoughts of their own mortality to the sheer admiration of knowing that someone cared enough to help others learn about the body, even in death. Poems or stories from medical students are often moving and heartfelt, and warrant being shared. PMID- 19217070 TI - Reduced N-acetylaspartate in the basal ganglia of a patient with anti-NMDA receptor encephalitis. PMID- 19217071 TI - Otic mesenchyme expression of Cre recombinase directed by the inner ear enhancer of the Brn4/Pou3f4 gene. AB - Brn4/Pou3f4 is a POU-domain transcription factor expressed in the otic mesenchyme that is required for the normal development of the inner ear. In this report, we describe the isolation of an otic mesenchyme enhancer in the Brn4 gene. Subsequently, this enhancer was used to drive the expression of Cre recombinase in the otic mesenchyme of transgenic mice. When intercrossed with the ROSA reporter strain, R26R, ss-galactosidase expression is detected in several inner ear structures derived from otic mesenchyme, including the temporal bone, spiral ligament, spiral limbus, and mesenchyme underlying sensory epithelium of the utricle, saccule and semicircular canals. Thus, this Cre pedigree can induce conditional rearrangement of genes in the otic mesenchyme, and will serve as a powerful genetic tool to characterize the function of genes in the mesenchymal tissues of the inner ear. PMID- 19217072 TI - Pathways to intrusive memories in a trauma analogue paradigm: a structural equation model. AB - BACKGROUND: Structural equation modeling was used to develop and test an integrated model of how emotional and cognitive factors contribute to the development of intrusive memories after a distressing event. METHODS: The emotional constructs included preexisting emotion and emotional reactivity to the distressing event. Cognitive constructs included peritraumatic cognitive processing and maladaptive coping strategies. RESULTS: Using a prospective design, 148 undergraduate students viewed a distressing film and recorded their film-related intrusive memories over the following week. RESULTS: The results indicated that emotional reactivity and, to a lesser extent, preexisting emotion, predicted peritraumatic cognitive processing during the film, which in turn predicted the use of maladaptive coping strategies (i.e., rumination, thought suppression, and safety behaviors). Furthermore, emotional reactivity and maladaptive coping strategies directly predicted intrusive memories. CONCLUSIONS: The observed model supported the importance of cognitive variables central to cognitive models, placing them in a broader context. PMID- 19217073 TI - Corticotropin-releasing factor-1 receptor activation mediates nicotine withdrawal induced deficit in brain reward function and stress-induced relapse. AB - BACKGROUND: Tobacco addiction is a chronic brain disorder that is characterized by a negative affective state upon smoking cessation and relapse after periods of abstinence. Previous research has shown that blockade of corticotropin-releasing factor (CRF) receptors with a nonspecific CRF1/CRF2 receptor antagonist prevents the deficit in brain reward function associated with nicotine withdrawal and stress-induced reinstatement of extinguished nicotine-seeking in rats. The aim of these studies was to investigate the role of CRF1 and CRF2 receptors in the deficit in brain reward function associated with precipitated nicotine withdrawal and stress-induced reinstatement of nicotine-seeking. METHODS: The intracranial self-stimulation (ICSS) procedure was used to assess the negative affective state of nicotine withdrawal. Elevations in brain reward thresholds are indicative of a deficit in brain reward function. Stress-induced reinstatement of nicotine seeking was investigated in animals in which responding for intravenously infused nicotine was extinguished by substituting saline for nicotine. RESULTS: In the ICSS experiments, the nicotinic receptor antagonist mecamylamine elevated the brain reward thresholds of the nicotine-dependent rats but not those of the control rats. The CRF1 receptor antagonist R278995/CRA0450 but not the CRF2 receptor antagonist astressin-2B prevented the elevations in brain reward thresholds associated with precipitated nicotine withdrawal. Furthermore, R278995/CRA0450 but not astressin-2B prevented stress-induced reinstatement of extinguished nicotine-seeking. Neither R278995/CRA0450 nor astressin-2B affected operant responding for chocolate-flavored food pellets. CONCLUSIONS: These studies indicate that CRF(1) receptors but not CRF(2) receptors play an important role in the anhedonic-state associated with acute nicotine withdrawal and stress induced reinstatement of nicotine-seeking. PMID- 19217074 TI - Co-administration of a D-amino acid oxidase inhibitor potentiates the efficacy of D-serine in attenuating prepulse inhibition deficits after administration of dizocilpine. AB - BACKGROUND: D-Serine, an endogenous agonist of the N-methyl-D-aspartate (NMDA) receptors, is effective in the treatment of schizophrenia. However, orally administered D-serine is metabolized substantially by D-amino acid oxidase (DAAO), diminishing its oral bioavailability. In this study, we examined the effects of oral D-serine administration with or without a DAAO inhibitor, 5 chloro-benzo[d]isoxazol-3-ol (CBIO), on the prepulse inhibition (PPI) deficits after administration of the NMDA receptor antagonist dizocilpine. METHODS: Vehicle or D-serine (30, 300, or 900 mg/kg) with or without CBIO (30 mg/kg) was orally administered to mice 1 hour before administration of dizocilpine (.1 mg/kg), and then the PPI of the acoustic startle response was measured. We measured the extracellular levels of D-serine in the frontal cortex after oral administration of D-serine with or without CBIO. RESULTS: Coadministration of CBIO with D-serine (30 mg/kg), but not D-serine (30 mg/kg) alone, significantly attenuated dizocilpine-induced PPI deficits. Furthermore, coadministration of CBIO significantly increased the extracellular levels of D-serine in the frontal cortex after administration of D-serine. CONCLUSIONS: These findings suggest that coadministration of CBIO significantly enhanced the efficacy of D-serine in attenuating PPI deficits by administration of dizocilpine. Therefore, coadministration of D-serine and a DAAO inhibitor has therapeutic potential for the treatment of schizophrenia. PMID- 19217075 TI - Cortisol response to stress in female youths exposed to childhood maltreatment: results of the youth mood project. AB - BACKGROUND: Few studies have examined stress reactivity and its relationship to major depressive disorder (MDD) and posttraumatic stress disorder (PTSD) among maltreated youth. We examined differences between maltreated and control participants in heart rate and cortisol resting and reactivity levels in response to a psychosocial stressor. METHODS: We recruited 67 female youths aged 12 to 16 with no prior history of depression from child protection agencies and a control group of 25 youths matched on age and postal code. Child maltreatment was measured with two self-report instruments. Psychiatric status was assessed using the Schedule for Affective Disorders and Schizophrenia for School-Aged Children. RESULTS: Piecewise multilevel growth curve analysis was used to model group differences in resting and reactivity cortisol levels and heart rate in response to the Trier Social Stress Test (TSST). During the resting period, both the maltreated and control groups showed a similar decline in levels of cortisol. During the reactivity phase, youth in the control group showed an increase in cortisol levels following the TSST and a gradual flattening over time; maltreated youth exhibited an attenuated response. This blunted reactivity was not associated with current symptoms of MDD or PTSD. There were no group differences in resting and reactivity levels of heart rate. CONCLUSIONS: These findings provide further support for hypothalamic-pituitary-adrenal axis dysregulation among maltreated youth. Since the ability to respond to acute stressors by raising cortisol is important for health, these findings may assist in understanding the vulnerability of maltreated youth to experience physical and mental health problems. PMID- 19217076 TI - Increased anxiety during anticipation of unpredictable aversive stimuli in posttraumatic stress disorder but not in generalized anxiety disorder. AB - BACKGROUND: Uncontrollability and unpredictability are key concepts related to re experiencing, avoidance, and hypervigilance symptoms of posttraumatic stress disorder (PTSD). However, little is known about the differential sensitivity of PTSD individuals to unpredictable stressors, relative to either healthy individuals or individuals with other anxiety disorders. This study tested the hypothesis that elevated anxious reactivity, specifically for unpredictable aversive events, is a psychophysiological correlate of PTSD. METHODS: Sixteen patients with PTSD (34.5 +/- 12.4 years) were compared with 18 patients with generalized anxiety disorder (GAD) (34.0 +/- 10.5 years) and 34 healthy control subjects (30.2 +/- 8.5 years). Participants were exposed to three conditions: one in which predictable aversive stimuli were signaled by a cue, a second in which aversive stimuli were administered unpredictably, and a third in which no aversive stimuli were anticipated. Startle magnitude was used to assess anxious responses to the threat cue and to contexts associated with each condition. RESULTS: Posttraumatic stress disorder and GAD patients showed normative enhancement of fear to the predictable threat cue, but the PTSD group displayed elevated anxiety during the unpredictable condition compared with participants with GAD and healthy control subjects. CONCLUSIONS: Anxious reactivity to unpredictable aversive events was heightened in PTSD but not in GAD and healthy subjects. Prior works also found signs of increased reactivity to unpredictable threat in panic disorder (PD), suggesting that PTSD and PD may involve shared vulnerability. As such, the current results inform understandings of classification, pathophysiology, and psychopharmacology of anxiety disorders, generally, and PTSD and panic disorder specifically. PMID- 19217077 TI - A double-blind, placebo-controlled study of the opiate antagonist, naltrexone, in the treatment of kleptomania. AB - BACKGROUND: Kleptomania is a rare psychiatric disorder characterized by recurrent stealing and for which there exists no empirically validated treatments. This study examined the efficacy and tolerability of the opioid antagonist naltrexone in adults with kleptomania who have urges to steal. METHODS: An 8-week, double blind, placebo-controlled trial was conducted to evaluate the safety and efficacy of oral naltrexone for kleptomania. Twenty-five individuals with DSM-IV kleptomania were randomized to naltrexone (dosing ranging from 50 mg/day to 150 mg/day) or placebo. Twenty-three subjects (92%) completed the study. Subjects were assessed every 2 weeks with the Yale Brown Obsessive Compulsive Scale Modified for Kleptomania (K-YBOCS), the urge and behavior subscales of the K YBOCS, the Kleptomania Symptom Assessment Scale (K-SAS), the Clinical Global Impressions Scale (CGI), and measures of depression, anxiety, and psychosocial functioning. RESULTS: Subjects assigned to naltrexone had significantly greater reductions in K-YBOCS total scores (p = .001), stealing urges (p = .032), and stealing behavior (p < .001) compared with subjects on placebo. Subjects assigned to naltrexone also had greater improvement in overall kleptomania severity (reflected in the CGI scores) (p < .001). The mean effective dose of naltrexone was 116.7 (+/-44.4) mg/day. CONCLUSIONS: Naltrexone demonstrated statistically significant reductions in stealing urges and behavior in kleptomania. Naltrexone was well tolerated. PMID- 19217078 TI - Ambience and drug choice: cocaine- and heroin-taking as a function of environmental context in humans and rats. AB - BACKGROUND: We have recently observed an unforeseen dissociation in the effect of environmental context on heroin versus cocaine self-administration in rats. Rats housed in the self-administration chambers (Residents) took more heroin than rats that were transferred to the self-administration chambers only for the test sessions (Nonresidents). The contrary was found for cocaine. The twofold aim of the present study was to investigate: 1) drug choice as a function of ambience in rats given access to both cocaine and heroin, and 2) ambience of choice for cocaine- versus heroin-taking in human addicts. METHODS: Resident and Nonresident rats with double-lumen intrajugular catheters were trained to self-administer cocaine (400 microg/kg/infusion) and heroin (25 microg/kg/infusion) on alternate days and then given the opportunity to choose between the two drugs during seven daily sessions. In the human study, we asked heroin and cocaine abusers where they preferred to take these drugs. RESULTS: Approximately 46.7% of Resident rats exhibited a preference for heroin over cocaine; 33.3% preferred cocaine, and 20% expressed no preference. In contrast, only 8.3% of Nonresident rats preferred heroin, whereas 66.7% preferred cocaine, and 25% expressed no preference. In the human study, 73% of co-abusers reported that they used heroin exclusively or mostly at home (22% used it outside the home), whereas only 25% reported using cocaine at home (67% took it outside their homes). CONCLUSIONS: Environmental context plays an important role in drug choice in both humans and rats self administering heroin and cocaine. PMID- 19217080 TI - Preparation and conformational analysis of C-glycosyl beta(2)- and beta/beta(2) peptides. AB - Ten C-glycosyl beta(2)- and beta/beta(2)-peptides with three to eight amino acid residues have been prepared. Solution and solid-phase peptide syntheses were employed to assemble beta(2)-amino acids in which C-glycosylic substituents are attached to the C-2 position of beta-amino acids. Conformational analysis of the C-glycosyl beta(2)-peptides using NMR and CD spectra indicates that the tripeptide can form a helical secondary structure. Besides, helix directions of the C-glycosyl beta/beta(2)-peptides are governed by the configuration at the alpha-carbon of the peptide backbone that originates from the stereocenter of the C-glycosyl beta(2)-amino acids. PMID- 19217079 TI - Pro-opiomelanocortin gene variation related to alcohol or drug dependence: evidence and replications across family- and population-based studies. AB - BACKGROUND: Opioidergic neurotransmission is critical in many, possibly all, forms of substance dependence. Several opioid-system genes have been shown to be associated with substance dependence disorders. The pro-opiomelanocortin gene (POMC) encodes several peptides important for endogenous opioidergic neurotransmission. We tested whether POMC genetic variation affects risk for substance dependence. METHODS: Five single nucleotide polymorphisms spanning POMC were examined in independent family and case-control samples. Family-based studies included 854 subjects from 319 African American (AA) families and 761 subjects from 313 European American (EA) families. Each family had a pair of siblings affected with cocaine and/or opioid dependence. Case-control studies included 791 cases (455 AAs and 336 EAs) affected with alcohol, cocaine, and/or opioid dependence and 682 control subjects (199 AAs and 483 EAs). RESULTS: Family based analyses revealed an association of rs6719226 with opioid dependence in AA families and rs6713532 with cocaine dependence in EA families (p = .010-.044). Case-control analyses demonstrated an association of rs6713532 with alcohol or cocaine dependence in EAs (p(allele-wise) = .003-.008). Moreover, the minor allele of rs1866146 was found to be a risk factor for cocaine or opioid dependence in AAs (p(allele-wise) = .010-.017) and for alcohol, cocaine, or opioid dependence in EAs (p(allele-wise) = .001-.003). Logistic regression analyses in which sex and age were considered and population stratification analyses confirmed these findings. Additionally, specific haplotypes increased risk for cocaine dependence (p = .023) in AAs and opioid dependence (p = .012) in EAs. CONCLUSIONS: Given these replicated results, we conclude that variation in POMC confers vulnerability to multiple forms of substance dependence. PMID- 19217081 TI - Glycoside modification of oleanolic acid derivatives as a novel class of anti osteoclast formation agents. AB - Oleanolic acid, a natural product, possesses an anti-osteoclast formation activity. Targeting at discovery of novel and potent anti-bone resorption agents, 22 glycosides of oleanolic acid derivatives (including d-galactopyranosides, d glucopyranosides, d-xylopyranoses, d-arabopyranoses and d-glycuronic acids) were synthesized at phase-transfer-catalyzed conditions (K(2)CO(3), Bu(4)NBr, CH(2)Cl(2)-H(2)O) and their inhibitory activity on the formation of osteoclast like multinucleated cells (OCLs) induced by 1alpha, 25-dihydroxy vitamin D(3) was evaluated in a co-culture assay system. The structure-activity relationships of these compounds were also discussed. PMID- 19217082 TI - A novel supramolecular compound 2,2'-bipyridyl-phosphotungstic acid: synthesis and catalysis. AB - A new supramolecular compound (C(10)H(8)N(2))(3.2).H(3)PW(12)O(40).25.6H(2)O (Bipy-PW(12)) was synthesized by self-assembly design, and characterized by elemental analysis, Fourier-transform infrared spectra (FTIR), and (31)P NMR spectra. Bipy-PW(12) can effectively catalyze oxidative degradation of chitosan with H(2)O(2) in heterogeneous phase. To obtain water-soluble chitosan with an average molecular weight of 5000, the optimum reaction conditions were determined as follows: reaction temperature, 80 degrees C; reaction time, 13min; H(2)O(2) concentration, 2.7mol/L; and mass ratio of Bipy-PW(12) to chitosan, 0.01. PMID- 19217083 TI - Synthesis of fused pyran-carbahexopyranoses as glycosidase inhibitors. AB - Synthesis of polyhydroxylated oxabicyclo[4,4,0]decanes, which constitute a new family of annulated carbasugars, has been accomplished in a stereoselective manner by employing readily available 1,2-anhydro-3,4,6-tri-O-benzyl-alpha-d glycopyranoses. PMID- 19217084 TI - 4-1BBL costimulation retrieves CD28 expression in activated T cells. AB - Binding of CD80/86 to CD28 is regarded as the main T cell costimulatory interaction. However, CD28 downregulates soon after T cell activation. To investigate potential cross-interaction between CD137 (4-1BB) and CD28, we stimulated T cells with anti-CD3 in the presence of A549 lung carcinoma cells expressing CD80/CD86 and 4-1BBL molecules, transduced into the cells using recombinant non-replicating adenoviruses. Following initial T cell proliferation, the proportion of CD28(+) cells in both CD4(+) and CD8(+) populations was rapidly reduced by CD80/86 costimulation, whereas cultures costimulated with just 4-1BBL continued to express CD28. CD28 was also downregulated in cultures costimulated with both CD80/86 and 4-1BBL. Interestingly, in cells costimulated with CD80/86 that had downregulated CD28 expression and ceased to proliferate, reactivation of proliferation by 4-1BBL costimulation also restored their CD28 expression. These findings show a positive effect of CD137 signalling on CD28 expression, similar to the effect of CD28 engagement on 4-1BB expression during the initial phases of T cell activation. Moreover, they point to the importance of signals through 4 1BB for the purposes of ex-vivo T cell activation and expansion. PMID- 19217085 TI - Familiarity effects on categorization levels of faces and objects. AB - It is well established that faces, in contrast to objects, are categorized as fast or faster at the individual level (e.g., Bill Clinton) than at the basic level (e.g., human face). This subordinate-shift from basic-level categorization has been considered an outcome of visual expertise with processing faces. However, in the present study we found that, similar to familiar faces, categorization of individually-known familiar towers is also faster at the individual level than at the basic-level in naive participants. In addition, category-verification of familiar stimuli, at basic and superordinate levels, was slower and less accurate compared to unfamiliar stimuli. Thus, the existence of detailed semantic information, regardless of expertise, can induce a shift in the default level of object categorization from basic to individual level. Moreover, the individually-specific knowledge is not only more easily-retrieved from memory but it might also interfere with accessing more general category information. PMID- 19217086 TI - Too much control can hurt: a threaded cognition model of the attentional blink. AB - Explanations for the attentional blink (AB; a deficit in identifying the second of two targets when presented 200-500 ms after the first) have recently shifted from limitations in memory consolidation to disruptions in cognitive control. With a new model based on the threaded cognition theory of multi-tasking we propose a different explanation: the AB is produced by an overexertion of control. This overexertion is produced by a production rule that blocks target detection during memory consolidation. In addition to fitting many known effects in the literature, the model predicts that adding certain secondary tasks will decrease the AB. In Experiment 1, a secondary task is added to the AB task in which participants have to respond to a moving dot. As predicted, AB decreases. Experiment 2 expands this result by controlling for learning, and adds a second variation, rotating the first target. For this variation the model predicts an increase in AB, which is indeed what we found. PMID- 19217087 TI - Dynamic photorefraction system: an offline application for the dynamic analysis of ocular focus and pupil size from photorefraction images. AB - Eccentric photorefraction is an optical technique used to assess static and/or dynamic changes in ocular focus (accommodation), ocular alignment (vergence) and pupil size. In this paper, we have developed and tested an offline application namely the dynamic photorefraction system (DPRS) which allows an accurate analysis of accommodation and pupil size from eccentric photorefraction images. The application uses the Microsoft componentized technology known as the Component Object Model (COM), includes distinct libraries for importing photorefraction videos and provides an accurate analysis and output of pupil size and accommodation. In addition, the system can interface with any custom built photorefractor allowing a widespread application in vision science experiments involving simultaneous measures of ocular focus and pupil size. PMID- 19217088 TI - Dual-phase lag effects on thermal damage to biological tissues caused by laser irradiation. AB - A dual-phase lag (DPL) bioheat conduction model, together with the broad beam irradiation method and the rate process equation, is proposed to investigate thermal damage in laser-irradiated biological tissues. It is shown that the DPL bioheat conduction model could predict significantly different temperature and thermal damage in tissues from the hyperbolic thermal wave and Fourier's heat conduction models. It is also found that the DPL bioheat conduction equations can be reduced to the Fourier heat conduction equations only if both phase lag times of the temperature gradient (tau(T)) and the heat flux (tau(q)) are zero. This is different from the DPL model for pure conduction materials, for which it can be reduced to the Fourier's heat conduction model provided that tau(q)=tau(T). Effects of laser parameters and blood perfusion on the thermal damage simulated in tissues are also studied. The result shows that the overall effects of the blood flow on the thermal response and damage are similar to those of the time delay tau(T). PMID- 19217089 TI - 3D MDCT-based system for planning peripheral bronchoscopic procedures. AB - The diagnosis and staging of lung cancer often begins with the assessment of a suspect peripheral chest site. Such suspicious peripheral sites may be solitary pulmonary nodules or other abnormally appearing regions of interest (ROIs). The state-of-the-art process for assessing such peripheral ROIs involves off-line procedure planning using a three-dimensional (3D) multidetector computed tomography (MDCT) chest scan followed by bronchoscopy with an ultrathin bronchoscope. We present an integrated computer-based system for planning peripheral bronchoscopic procedures. The system takes a 3D MDCT chest image as input and performs nearly all operations automatically. The only interaction required by the physician is the selection of ROI locations. The system is computationally efficient and fits smoothly within the clinical work flow. Integrated into the system and described in detail in the paper is a new surface definition method, which is vital for effective analysis and planning to peripheral sites. Results demonstrate the efficacy of the system and its usage for the live guidance of ultrathin bronchoscopy to the periphery. PMID- 19217090 TI - Estrogen and progesterone receptors in smooth muscle component of deep infiltrating endometriosis. AB - OBJECTIVE: To analyze the expression of estrogen (ER) and progesterone (PR) receptors in the smooth muscle component (SMC) of deep infiltrating endometriosis (DIE). DESIGN: A prospective clinical and pathologic study of 60 cases of DIE. SETTING: University Hospital Department of Gynacology. PATIENT(S): Sixty patients with symptomatic DIE (uterosacral endometriosis n = 14; bladder endometriosis n = 10; colonic endometriosis n = 16; rectovaginal endometriosis n = 20). INTERVENTION(S): Laparoscopic surgery. MAIN OUTCOME MEASURE(S): The expression of ER and PR was studied by immunohistochemistry in the SMC directly around endometriotic foci and at distance (at least >1.5 cm) from them in correlation with proliferative and secretory phases of cycle. RESULTS: The ER and PR were present in the SMC of DEI in each location excepting colonic endometriosis where ER were absent. Independently of cycle's phases the PR were more abundant than ER. With the exception of rectovaginal endometriosis, where the ER and PR were more abundant in the proliferative than in the secretory phase, in other locations the ER and PR did not differ significantly with cycle's phases. Last, if ER and PR were more abundant in SMC around endometriotic foci than at a distance from them. However, the difference was not significant. CONCLUSIONS: Our data substantially confirm for the first time that in various forms of DIE, ER and PR are present not only in glands and stroma but also in the smooth muscle major histologic component of this disease. PMID- 19217091 TI - Human embryo twinning with applications in reproductive medicine. AB - OBJECTIVE: To assess the efficacy of human embryo twinning by blastomere biopsy at different early embryonic stages (splitting efficiency) and to determine the in vitro developmental capacity of twinned human embryos (developmental efficiency). DESIGN: Randomized comparative study. SETTING: Private IVF centers. PATIENT(S): Couples undergoing IVF donating triploid embryos. INTERVENTION(S): Embryos at the 2- to 5- and 6- to 8-cell stage were split into twin embryos. Half the number of blastomeres from donor embryos were removed and inserted into recipient empty zonae pellucidae. After embryo splitting, donor and recipient embryos were cultured in vitro. MAIN OUTCOME MEASURE(S): Development of twinned embryos to the blastocyst stage. RESULT(S): The number of developing embryos obtained after splitting could be increased in comparison with the number of embryos available before splitting at the 6- to 8-cell stage but not at the 2- to 5-cell stage (splitting efficiency). Splitting of 6- to 8-cell embryos yielded superior rates of twin embryos developing to blastocysts (developmental efficiency). Twinning success was related to the superior morphological quality of embryos used for splitting. CONCLUSION(S): This is the first report on twinned human embryos developing to blastocysts. This study exhibits the potential for novel applications in human assisted reproduction. PMID- 19217092 TI - Nitric oxide platelet production in spontaneous miscarriage in the first trimester. AB - OBJECTIVE: To investigate the role played by platelet nitric oxide (NO) metabolism in patients with spontaneous miscarriage (SM) and recurrent spontaneous miscarriage (RSM) compared with healthy pregnant women. DESIGN: Retrospective case-control study. SETTING: Patients and controls in an academic research environment. INTERVENTION(S): None. PATIENT(S): Thirty singleton pregnant women who experienced SM, nine singleton pregnant women who presented with RSM, and 30 singleton healthy pregnant women matched for age, parity, and gestational age were enrolled. MAIN OUTCOME MEASURE(S): NO levels and peroxynitrite (ONOO(-)) production; moreover, inducible NO synthase (iNOS), endothelial NO synthase (eNOS), and nitrotyrosine expression (N-Tyr) were observed in the same samples. RESULT(S): A significant increase was shown in platelet NO and ONOO(-) levels and in iNOS and N-Tyr both in SM and in RSM pregnant women compared with controls. CONCLUSION(S): The data herein reported imply that a modified NO pathway might play a key role in the physiological changes of advancing gestation but may also contribute to the pathophysiology of spontaneous miscarriage. Thus, any factors balancing NO metabolism might be useful in the treatment of miscarriage, thus reducing the substantial morbidity and associated mortality. PMID- 19217093 TI - Predictors of urinary albumin excretion in women with polycystic ovary syndrome. AB - OBJECTIVE: To evaluate urinary albumin excretion (UAE) in normotensive and nondiabetic women with polycystic ovary syndrome (PCOS) in relation to their clinical, endocrine, and metabolic profiles. DESIGN: Observational study. SETTING: University fertility center. PATIENT(S): Sixty-three women with PCOS were evaluated. INTERVENTION(S): Clinical assessments and urine and blood testing. MAIN OUTCOME MEASURE(S): Urinary albumin excretion, systolic and diastolic blood pressure, serum levels of LH, FSH, PRL, T, 17 hydroxyprogesterone, glucose, insulin, lipids, C-reactive protein, and 24-hour urinary free cortisol. RESULT(S): In univariate and multivariate correlation analysis, UAE correlated with diastolic blood pressure, insulin area under the curve during glucose tolerance test, PRL, and 17-hydroxyprogesterone. Overt microalbuminuria was detected in a significant proportion of subjects. CONCLUSION(S): Urinary albumin excretion in women with PCOS correlates well with other cardiovascular risk factors. Because the relationship between UAE and adverse cardiovascular events is continuous, evaluation of UAE in the presence of PCOS may provide clinically relevant information and may aid in selecting appropriate patients for more aggressive treatment of likely aggravating factors, such as hyperinsulinemia or borderline hypertension. PMID- 19217094 TI - Fertility status in male cystinosis patients treated with cysteamine. AB - OBJECTIVE: To analyze the fertility status in adult, male cystinosis patients treated with cysteamine. Cystinosis is an autosomal recessive disease leading to intralysosomal cystine accumulation. Worldwide, a few female cystinosis patients have given birth. However, no male cystinosis patients are known to have induced pregnancy. Adequate cysteamine treatment might improve male fertility. PATIENT(S): Seven male cystinosis patients (19-43 years) were submitted. INTERVENTION(S): Glomerular filtration rate was estimated using the Cockcroft formula. Serum LH, FSH, testosterone, and inhibin B were determined. Semen analysis was performed in five patients. Testicular biopsy was performed in one patient. RESULTS: Glomerular filtration rate ranged between 10 and 110 (normal >90) mL/min/1.73 m(2), LH and FSH levels ranged between 7.4 and 235.0 (normal 1.4 8.5) E/L and 6.8-298.0 (normal 1.5-11) E/L, respectively. Plasma testosterone level ranged between 8.7 and 31.3 (normal 11-45) nmol/L; plasma inhibin B level ranged between 10 and 210 (normal 150-400) ng/L. All of the collected sperm samples showed azoospermia. The testicular biopsy showed a Johnson score of 8 to 9. CONCLUSION(S): We demonstrate azoospermia in male cystinosis patients, even if adequately treated with cysteamine starting from an early age. The finding of spermatogenesis in the testis biopsy of one patient may provide opportunities to male cystinosis patients to produce their own offspring by in vitro fertilization after testicular sperm extraction. PMID- 19217095 TI - Comparing anti-Mullerian hormone (AMH) and follicle-stimulating hormone (FSH) as predictors of ovarian function. AB - We compared predictive values of anti-Mullerian hormone (AMH) and baseline FSH with respect to IVF cycle outcomes based on oocyte numbers retrieved and number of clinical pregnancies established. In 76 IVF cycles investigated, AMH was clearly superior in predicting IVF outcomes in comparison with FSH. PMID- 19217096 TI - Cellular senescence in usual type uterine leiomyoma. AB - OBJECTIVE: To evaluate the role of senescence in symptomatic patients with multifibroids. DESIGN: A cohort study. SETTING: University research laboratory. PATIENT(S): Eighty-six fibroids collected from 14 patients who underwent myomectomy or hysterectomy. INTERVENTION(S): Senescence-associated beta galactosidase (SA-beta-Gal) stain in fresh-frozen tissue; reverse-transcription polymerase chain reaction (RT-PCR); MicroRNA in situ hybridization (MISH); immunohistochemistry in formalin-fixed paraffin-embedded tissue. MAIN OUTCOME MEASURE(S): Senescence measured by percentage of SA-beta-Gal-positive cells; levels of let-7 microRNAs measured by RT-PCR and MISH; expression of p16(INK4a), Ki-67, HMGA1, and HMGA2 scaled by immunoreactivity. RESULT(S): About 58% (48 of 82) of tumors showed significant senescent change (SA-beta-Gal positive) in >10% of the tumor volume. The overall trend was a higher level of senescence in small fibroids and older-aged women. Senescent fibroids were additionally shown to have, high levels of let-7 c, d, and f-2 and a low Ki-67 index. CONCLUSION(S): Senescence is a common cellular change in a large proportion of usual type fibroids. Similarly, senescence may explain the variation in growth rates of these tumors, and may prove to be an important molecular and cellular target in prevention of fibroid growth. PMID- 19217097 TI - Incidence of monozygotic twins in blastocyst and cleavage stage assisted reproductive technology cycles. AB - OBJECTIVE: To study the incidence of monozygotic twins (MZT) in blastocyst and cleavage stage ET. DESIGN: Retrospective review. PATIENT(S): Four hundred ninety six women undergoing IVF/intracytoplasmic sperm injection cycles at a private assisted reproductive technology (ART) center. INTERVENTION(S): Patients undergoing ART were divided according to the stage of ET into blastocyst transfer (BT) and cleavage stage (CS) ET. MAIN OUTCOME MEASURE(S): Incidence of MZT as noted on vaginal ultrasound at 6 to 8 weeks. RESULT(S): There were 374 (75.4%) BT cycles, and 122 (24.6%) CS cycles. Women in the BT group were significantly younger, had more oocytes retrieved, and had less embryos transferred compared with the CS group. The clinical pregnancy rate was significantly higher in the BT group at 67.9% (254 of 374), compared with 37.7% (46 of 122) among the CS group. There was a significantly higher incidence of multiple gestation in the BT group compared with CS group (37.4% compared with 19.6%). The overall incidence of MZT was 1.3%, but differed with the stage of ET: the incidence of MZT was 1.57% (4 of 254) in the BT group, and 0 (0 of 46) in the CS group. CONCLUSION(S): Contrary to the older published literature on MZT in BT cycles, the incidence of MZT is low. Women undergoing ART therefore should not be discouraged to undergo BT for fear of MZT, especially in light of the higher pregnancy rate and lower number of transferred embryos noted in those cycles compared with cleavage stage transfers. PMID- 19217098 TI - Chronic endometritis is a frequent finding in women with recurrent implantation failure after in vitro fertilization. AB - OBJECTIVE: To determine the role of endometrial sampling for identification and treatment of chronic endometritis (CE) in patients undergoing IVF-ET who repeatedly failed to conceive despite the transfer of good-quality embryos. DESIGN: Retrospective chart review. SETTING: University-based tertiary fertility center. PATIENT(S): Thirty-three patients with recurrent implantation failure (RIF) who underwent endometrial sampling and subsequent ET were analyzed based on immunohistochemically confirmed CE: CE present on biopsy (group 1; n = 10) and CE absent on biopsy (group 2; n = 23). Patients with RIF undergoing IVF cycles during the same time period who did not have endometrial sampling were used as controls (group 3; n = 485). INTERVENTION(S): Endometrial sampling for CE and subsequent antibiotic treatment in affected patients followed by another IVF-ET cycle. RESULT(S): Chronic endometritis was identified in 30.3% of patients with RIF. Group 1 had lower implantation rates (11.5%) in the IVF cycle following treatment than did group 2 and group 3 (32.7% and 20.3%, respectively). Clinical pregnancy and ongoing pregnancy rates were similar across groups. CONCLUSION(S): Recurrent implantation failure warrants investigation of CE as a contributing factor. Women demonstrating CE on endometrial sampling have lower implantation rates in a subsequent IVF-ET cycle; however, there were no differences in subsequent clinical pregnancy or ongoing pregnancy rates after successful antibiotic treatment. PMID- 19217099 TI - Vitrification of oocytes produces high pregnancy rates when carried out in fertile women. AB - OBJECTIVE: To determine the efficiency of our vitrification technique when applied in young fertile women. DESIGN: Clinical research and application. SETTING: In vitro fertilization center. PATIENT(S): Twenty-one women were recruited from the navy community with 19 patients finishing the study. INTERVENTION(S): Vitrified oocytes with use of the electron microscopic grid method were warmed 6 months after vitrification. Surviving metaphase II oocytes were microinjected for fertilization, and clinical results were evaluated. MAIN OUTCOME MEASURE(S): Survival, fertilization, and cleavage rate. Pregnancy and implantation rate. RESULT(S): Three hundred ninety-five oocytes were warmed, of which 320 oocytes (81.0%) survived. Two hundred eighty-five metaphase II oocytes were microinjected for fertilization; 206 of them (72.3%) fertilized, and 53 embryos were transferred to 19 patients (in 20 warming cycles). Twenty-four of 53 transferred embryos (45.3%) implanted as confirmed by ultrasound examination. Of the 20 transfers, 16 resulted in clinical pregnancy (80%), 3 miscarried (15%), and 13 (65%) went on to produce 20 live births, respectively. This is much higher in comparison with our previous data using supernumerary oocytes where the rates of implantation and pregnancy were 6% and 21%. Live-birth rates per warmed oocyte and per injected oocyte were 5.1% and 7.2%, respectively. CONCLUSION(S): High pregnancy and implantation rates were observed after 6 months of cryopreservation by vitrification when oocytes from fertile woman were used. Proper screening of candidates for oocyte cryopreservation is of crucial importance to assure a favorable pregnancy outcome. PMID- 19217100 TI - Decreased sperm motility is associated with air pollution in Salt Lake City. AB - OBJECTIVE: To study the correlation between indices of air pollution and sperm parameters. DESIGN: Ecological study. SETTING: Contained geographic area. PATIENT(S): Resident men presenting for semen analysis or artificial insemination. INTERVENTION(S): Analysis of levels of particulate air pollution (particulate matter [PM] 2.5) over a five-year period in relation to sperm parameters obtained from semen analyses and separately in relation to sequences of sperm parameters at the time of semen preparations for artificial insemination. To account for the duration of spermatogenesis (72 days), "corrected" variables were created by shifting backward 1, 2, 3 and 4 months each of the semen parameters. The final analysis corrected for season of the year and current temperature. MAIN OUTCOME MEASURE(S): Sperm concentration, sperm motility and sperm morphology. RESULT(S): The study included 1,699 semen analyses and 877 inseminations. PM 2.5 levels were highest in the winter months, when the ambient temperature was the lowest. Semen analysis data showed that values of PM 2.5 were negatively correlated to sperm motility two months and three months following the recording of the PM 2.5 values. Artificial insemination data also showed that sperm motility correlated negatively with PM 2.5 values recorded three months previously. CONCLUSION(S): Both semen analysis and sperm parameters data obtained from men presenting for multiple inseminations over time showed that air pollution is associated with reduced sperm motility two to three month after exposure. PMID- 19217101 TI - Effects of rapid distraction rate on new bone formation during mandibular distraction osteogenesis in goats. AB - OBJECTIVE: Distraction osteogenesis typically requires a long treatment period, which can lead to bone and soft-tissue infection and considerable patient discomfort. Use of a rapid distraction rate in craniofacial distraction osteogenesis to shorten the distraction period is possible owing to the unique characteristics of craniofacial bones, including an abundant blood supply and rapid bone healing compared with long bones. The effects of using a rapid distraction rate in the treatment of craniofacial deformities are currently unclear, however. The objective of this study was to investigate the effects of a rapid distraction rate on new bone formation during mandibular distraction osteogenesis in goats. METHODS: Sixteen goats were randomly divided into four groups consisting of four goats each. In Groups A, B, and C, the right mandible of each goat was distracted at a rate of 0.8mm/d, 1.6mm/d, and 2.0mm/d, respectively; Group D was the control group and did not undergo distraction. Six weeks after the conclusion of distraction, bone densitometry and three-point bending testing were performed in all groups. RESULTS: The mean bone density value of goats in Group A was significantly higher than those of all the other groups (p<0.05), and the mean bone density value of goats in Group C was significantly lower than those of all the other groups (p<0.05). The mean curve slope, peak stress, bending modulus, and energy to failure values of Groups A, B, and C were all significantly lower than those of the control group (p<0.05). As the distraction rate increased, the curve slope and peak stress values gradually declined (p<0.05). CONCLUSIONS: Use of a rapid distraction rate in mandibular distraction osteogenesis may have detrimental effects on the quality of new bone, despite the abundant blood supply of craniofacial bones. PMID- 19217102 TI - W.S.B.--a fluoroscopy C-arm communication strategy. AB - BACKGROUND: Good communication between an operating clinician and the attending radiographer when using C-arm fluoroscopy is vital. Efficient and safe use of ionising radiation protects all theatre staff. The object of this study was to design and test a single communication language for use in all fluoroscopy assisted procedures. METHOD: A clinician guiding a radiographer performed a control 'no-rules' experiment of 56 targeting manoeuvres with a fluoroscopic C arm on a scientific model. A single beam laser replaced the usual ionising radiation beam. The time taken for each targeting along with the laser exposure required was recorded. Following a period of instruction, the experiment was repeated using only our pre-defined communication strategy. The data was collected for statistical analysis. RESULTS: A statistically significant reduction was seen in the time taken for each targeting manoeuvre and the laser exposure required when using our strategy. CONCLUSION: This study shows that using our pre-prepared communication strategy could significantly improve efficiency and safety of fluoroscopic C-arm use. PMID- 19217103 TI - Acute care of hip fractures in centenarians--do we need more resources? AB - The centenarian population in England and Wales is the most rapidly increasing age group, yet we have little information as regards their acute hospital stay and delay in surgery after hip fracture. We reviewed the records of 26 centenarians with hip fracture between 2000 and 2007 and compared them to a randomly selected control group of 50 hip fracture patients between the ages of 75 and 85 years. The mean stay in acute orthopaedic wards for centenarians was 20.7 days and for the control group was 14.9 days (p=0.015). Centenarians had a mean delay in surgery of 3.6 days while non-centenarians were operated within a mean of 1.9 days, which was not statistically significant (p=0.241). The longer acute hospital stay in our centenarian cohort would amount to a mean extra cost of pound 2511 per patient. PMID- 19217104 TI - Severe and multiple trauma in older patients; incidence and mortality. AB - OBJECTIVE: To examine the differences between severely injured older patients (aged over 65 years) compared with similarly injured younger adults in terms of incidence, inpatient mortality and factors predicting outcome. METHODS: Data prospectively entered into the Trauma Audit and Research Network (TARN) database from our level I trauma unit over a 5-year period were retrospectively examined, with 3172 patients included in the final analysis. RESULTS: Older patients accounted for 13.8% of those with severe injuries (Injury Severity Score 16 or more) and almost 2% of our trauma admissions overall. High energy injuries were responsible for the majority of these injuries though relatively minor trauma became increasingly important in older patients. Mortality rates in the older patients were more than twice those seen in the adult population (19% in the under 40's to almost 50% in the over 75's). Age, Injury Severity Score and Glasgow Coma Score continued to be predictive of mortality in older patients but other factors relevant in younger adults were not. CONCLUSIONS: Patients in the older group without physiological derangement on admission were still at a relatively high risk of inpatient mortality. This was in contrast to the younger patients, suggesting that it might be more difficult to predict which older patients might benefit from more aggressive monitoring or treatment. Despite increased mortality in older patients, significant survival rates were achieved even in the oldest. Active treatment should not be withdrawn on the basis of age alone. PMID- 19217105 TI - Effects of abdominal massage in management of constipation--a randomized controlled trial. AB - BACKGROUND: Associated with decreases in quality of life, constipation is a relatively common problem. Abdominal massage appears to increase bowel function, but unlike laxatives with no negative side effects. Because earlier studies have methodological flaws and cannot provide recommendations, more research is needed. OBJECTIVE: This study investigates the effects of abdominal massage on gastrointestinal functions and laxative intake in people who have constipation. DESIGN: Randomized controlled trial. PARTICIPANTS AND METHOD: A sample of 60 people with constipation was included and randomized in two groups. The intervention group received abdominal massage in addition to an earlier prescribed laxative and the control group received only laxatives according to earlier prescriptions. Gastrointestinal function was assessed with Gastrointestinal Symptoms Rating Scale (GSRS) on three occasions; at baseline, week 4 and week 8. The statistical methods included linear regression, Wilcoxon sign rank test, and Mann-Whitney U-test. RESULT: Abdominal massage significantly decreased severity of gastrointestinal symptoms assessed with GSRS according to total score (p=.003), constipation syndrome (p=.013), and abdominal pain syndrome (p=.019). The intervention group also had significant increase of bowel movements compared to the control group (p=.016). There was no significant difference in the change of the amount of laxative intake after 8 weeks. CONCLUSIONS: Abdominal massage decreased severity of gastrointestinal symptoms, especially constipation and abdominal pain syndrome, and increased bowel movements. The massage did not lead to decrease in laxative intake, a result that indicates that abdominal massage could be a complement to laxatives rather than a substitute. PMID- 19217106 TI - "Insufficient" but still "necessary"? EBPM's dangerous leap of faith: commentary on Porter and O'Halloran (2009). PMID- 19217107 TI - Psychometric evaluation of the Mainland Chinese version of the Edinburgh Postnatal Depression Scale. AB - BACKGROUND: The Edinburgh Postnatal Depression Scale (EPDS) has been validated in many countries, but not in Mainland China. OBJECTIVES: This study investigated the reliability and validity of the Mainland Chinese version of the EPDS. DESIGN: A three-stage design was used for this study. Stage I consisted of a multi stepped process of forward and backward translation, using a panel of six experts to test content validity. Stage II established the psychometric properties of the EPDS by examining the convergent, discriminant and construct validity, internal consistency and stability of the scale. Stage III established its sensitivity, specificity and the optimal cutoff score of the EPDS according the DSM-IV-TR criteria using the Structured Clinical Interview. SETTING: Three regional public hospitals in Chengdu. PARTICIPANTS: A convenience sample was composed of 312 and 451 pregnant women at 28-36 weeks' gestation. METHOD: Translation and back translation of the original English instrument and content validation by an expert panel. Receiver operating characteristic (ROC) analysis was carried out to evaluate the global functioning of the scale. The Beck Depression Inventory (BDI), Dyadic Adjustment Scale (DAS) and standard SF-12 Health Survey (SF-12) were used to investigate the convergent, discriminant and construct validity. The Cronbach's alpha reliability coefficient, split-half reliability and test-retest reliability were used to examine the internal consistency and stability of the scale. RESULTS: The translation process was rigorously conducted to ensure that equivalence was established. Content validity was confirmed by a satisfactory level of agreement with a content validity index (CVI) of 0.93. The area under curve (AUC) was 85.6% and the logistic estimate for the threshold score was 9.5 (sensitivity, 80.0%; specificity, 83.03%) for clinical depression. Convergent and construct validity was supported and discriminant validity suggested that the EPDS successfully discriminated among the non-depressed, mildly and clinically depressed groups. The split-half reliability of the EPDS was 0.76, Cronbach's alpha was 0.79 and test-retest reliability was 0.85. CONCLUSIONS: Based on the results of this psychometric testing, the Mainland Chinese version of the EPDS is considered ready for use in the routine screening of pregnant women. It is hoped that with this type of cross-cultural information, the universality of the constructs of the instrument can be reliably demonstrated. PMID- 19217108 TI - Independent protective roles for macrophage Abcg1 and Apoe in the atherosclerotic lesion development. AB - OBJECTIVE: ATP-binding cassette transporter G1 (Abcg1) and apolipoprotein E (Apoe) play a role in macrophage cholesterol efflux and consequently the development of atherosclerosis. A possible interaction between Abcg1 and Apoe in cholesterol efflux was postulated, but the potential combined action of these proteins on atherosclerotic lesion formation is unclear. METHODS: LDL receptor knockout (KO) mice were transplanted with bone marrow from Abcg1/Apoe double KO (dKO) mice, their respective single knockouts, and wild-type (WT) controls and challenged with a high-fat/high-cholesterol diet for 6 weeks to induce atherosclerosis. RESULTS: No differences were found in serum lipid levels. The mean atherosclerotic lesion area in dKO transplanted animals (187+/ 18x10(3)microm(2)) was 1.4-fold (p<0.01) increased compared to single knockouts (Abcg1 KO: 138+/-5x10(3)microm(2); Apoe KO: 131+/-7x10(3)microm(2)) and 1.9-fold (p<0.001) as compared to WT controls (97+/-15x10(3)microm(2)). In vitro cholesterol efflux experiments established that combined deletion of Abcg1 and Apoe leads to a larger attenuation of macrophage cholesterol efflux to HDL as compared to single knockouts. CONCLUSIONS: Single deletion of macrophage Abcg1 or Apoe does lead to a moderate non-significant increase in atherosclerotic lesion development as tested by ANOVA, while combined deletion of Abcg1 and Apoe induces a more dramatic and significant increase in atherosclerosis. Our results indicate an additive, independent effect for both macrophage Abcg1 and Apoe in the prevention of atherosclerosis. PMID- 19217110 TI - Optimization strategies for off-line two-dimensional liquid chromatography. AB - A step by step strategy of optimization of comprehensive off-line two-dimensional liquid chromatography (2D-LC) separations is proposed. The goal of an optimization process in the separation sciences is either to achieve a given resolution (a target peak capacity in 2D-LC) within as short a time as possible or to reach the highest possible resolution in a given analysis time. The proposed method takes into account the characteristics of the columns used in the first and the second dimension and the number of fractions of the first dimension eluent that should be collected. The effect of the time spent during the analysis on the second dimension column to carry out necessary tasks that are not the separation itself (called the additional time) on the maximum peak capacity that is achievable was carefully investigated. It was shown that (1) an increase in the peak capacity of the first dimension column combined with the collection of larger volume fractions permits a significant reduction of the time needed to achieve the desired peak capacity; and (2) there is an optimum fraction collection ratio (or number of collected fractions per peak) which yields the target peak capacity in the minimum time. The proposed strategy was used for the optimization of the separation of samples of BSA tryptic digest by an off-line 2D LC using an SCXmultiply sign in circleRP-HPLC method. As a result of this optimization, a peak capacity of 4000 could be achieved in about 5h with the two columns available. The time needed for the optimized analysis was less than two thirds of the analysis time that would have been needed had the conventional rule of thumb of sample collection in comprehensive on-line 2D-LC (4 samples/peak) been followed. PMID- 19217109 TI - Mechanisms of improved knee flexion after rectus femoris transfer surgery. AB - Rectus femoris transfer is frequently performed to treat stiff-knee gait in subjects with cerebral palsy. In this surgery, the distal tendon is released from the patella and re-attached to one of several sites, such as the sartorius or the iliotibial band. Surgical outcomes vary, and the mechanisms by which the surgery improves knee motion are unclear. The purpose of this study was to clarify the mechanism by which the transferred muscle improves knee flexion by examining three types of transfers. Muscle-actuated dynamic simulations were created of ten children diagnosed with cerebral palsy and stiff-knee gait. These simulations were altered to represent surgical transfers of the rectus femoris to the sartorius and the iliotibial band. Rectus femoris transfers in which the muscle remained attached to the underlying vasti through scar tissue were also simulated by reducing but not eliminating the muscle's knee extension moment. Simulated transfer to the sartorius, which converted the rectus femoris' knee extension moment to a flexion moment, produced 32+/-8 degrees improvement in peak knee flexion on average. Simulated transfer to the iliotibial band, which completely eliminated the muscle's knee extension moment, predicted only slightly less improvement in peak knee flexion (28+/-8 degrees ). Scarred transfer simulations, which reduced the muscle's knee extension moment, predicted significantly less (p<0.001) improvement in peak knee flexion (14+/-5 degrees ). Simulations revealed that improved knee flexion following rectus femoris transfer is achieved primarily by reduction of the muscle's knee extension moment. Reduction of scarring of the rectus femoris to underlying muscles has the potential to enhance knee flexion. PMID- 19217111 TI - Fundamental equation of the dual flow rate-solvent gradient elution in liquid chromatography. AB - An alternative expression of the fundamental equation of multi-mode gradient elution involving simultaneous changes in mobile phase composition and flow rate is derived using simple kinetic arguments and graphic interpretation. The new expression consists of a system of two integral equations and provides an easy and direct way of predicting retention times under dual-mode gradient conditions. PMID- 19217113 TI - Freely dissolved concentrations of anionic surfactants in seawater solutions: optimization of the non-depletive solid-phase microextraction method and application to linear alkylbenzene sulfonates. AB - A solid-phase microextraction method (SPME) has been optimized for the analysis of freely dissolved anionic surfactants, namely linear alkylbenzene sulfonates (LAS), in seawater. An effect of the thermal conditioning treatment on the polyacrylate fiber coating was demonstrated for both uptake kinetics and sorption isotherm linearity. Thermal conditioning at 120 degrees C yielded linear sorption isotherms and reproducible SPME measurements for several individual LAS compounds, with detection limits at the low microgram per liter range. Sorption of LAS to the conditioned SPME fiber was independent of LAS co-solutes in mixtures. The method has been applied to study the precipitation of LAS in seawater, and solubility data for a wide range of individual LAS constituents is presented for the first time. Hence, the developed SPME method for the anionic LAS has shown to be a useful tool in complex matrices. The advantage of the SPME analyses in complex matrices is, besides its simplicity, that it also leads to clean extracts for chromatographic analyses. PMID- 19217112 TI - Short monolithic columns for purification and fractionation of peptide samples for matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry analysis in proteomics. AB - This study records a novel application of methacrylate-based monolithic columns for MALDI-TOF/TOF MS analyses in proteomics for pre-concentration and separation of peptides derived from protein digestion. Reversed-phase monolithic capillary columns (30mm x 0.32mm i.d.) were created inside the fused silica capillary via thermal-initiated free-radical polymerization of ethylene glycol dimethacrylate and lauryl methacrylate monomers in the presence of 1-propanol and 1,4-butandiol as a porogen system. The elution of peptides was achieved using a linear gradient of acetonitrile from 0 to 60% in water with 0.1% trifluoroacetic acid formed in a microsyringe. Individual fractions of separated peptides were collected on the MALDI target spots covered with alpha-cyano-4-hydroxycinnamic acid used as a matrix and then they were analyzed using MALDI-TOF/TOF mass spectrometry. The developed method was tested with a mixture of tryptic peptides from bovine serum albumin and its applicability was also tested for tryptic in-gel digests from barley grain extracts of water soluble proteins separated using SDS gel electrophoresis. The number of detected peptides was approximately three to four times higher compared to the analysis without previous separation. These results show an improved quality of sample information with the higher amount of identified peptides which increased protein sequence coverage and improved sensitivity of mass spectrometry measurements. PMID- 19217114 TI - Electrospinning preparation, characterization and photocatalytic properties of Bi2O3 nanofibers. AB - Bi(2)O(3) nanofibers with diameter of 70-200 nm were successfully prepared by electrospinning a precursor mixture of polyacrylonitrile (PAN)/bismuth nitrate, followed by calcination treatment of the electrospun polymer/inorganic composite fibers. The resulting Bi(2)O(3) nanofibers were characterized with X-ray diffraction, X-ray photoelectron spectroscopy, scanning electron microscopy, FT IR spectra and UV-vis diffuse reflectance spectroscopy. The Bi(2)O(3) nanofibers calcined at 500 degrees C exhibited beta phase, and whereas the nanofibers calcined at 550 and 600 degrees C both exhibited dominant beta phase together with a trace amount of alpha phase. Photocatalytic experiments indicated that the obtained Bi(2)O(3) nanofibers calcined at 500 degrees C were highly active for photodegradation of organic pollutants Rhodamine B (RB). Furthermore, the Bi(2)O(3) nanofibers could be easily recycled without decrease of the photocatalytic activity. PMID- 19217115 TI - Normalization, comparison, and scaling of adsorption data: arsenate and goethite. AB - Various approaches for analyzing adsorption data were examined to determine the best method for reporting and interpreting the results of adsorption experiments and ultimately extrapolating laboratory measurements to the field. The interactions of arsenate and goethite were used as representative adsorbate and adsorbent, respectively, although the general principles are applicable to other adsorbate-adsorbent systems as well. A modeling exercise was conducted first to determine the theoretical principles governing the comparison and scaling of adsorption data. These principles were then tested on a suite of experimental data, both new and previously published. LogK(D) is significantly more sensitive to variations in adsorbate (As(T)) or adsorbent (Fe(T)) concentrations than either adsorbed concentration (q) or percentage adsorbed. The sensitivity of K(D) relative to q occurs due to the non-linearity of the adsorption isotherm at a given pH, since as the equilibrium aqueous concentration approaches zero, q also approaches zero while K(D) approaches infinity. Varying As(T) and Fe(T) while keeping As(T)/Fe(T) fixed yields more consistent values of percentage adsorbed, logK(D), and q, although the adsorbate-to-adsorbent ratios used in laboratory studies often have a rather narrow range compared to those possible in the field. Specific surface area is also a better scaling parameter than the mass of adsorbent, especially between systems with differing adsorbents with markedly different specific surface areas (e.g., natural versus synthetic goethite). Our results have significant implications to contaminant transport modeling, as the constant K(D) approach is the most common method of modeling contaminant transport, while contaminant concentrations in the field are typically low, precisely the conditions where K(D) is most sensitive. PMID- 19217116 TI - Electrochemical preparation and structural characterization of platinum thin film on a polypyrrole film modified ITO electrode. AB - Platinum nanostructures have been fabricated by electrochemical deposition of platinum onto indium tin oxide (ITO) glass substrate modified with thin polypyrrole film. The crystal size and the number density of the platinum nanoparticles have been examined by varying several deposition parameters, including the thickness of the PPy film and the current densities for platinum deposition. Optimal conditions for uniform growth of nanoparticles well-dispersed on the ITO have been determined, along with insight into the mechanism of crystal growth. The PPy film thickness principally affect the size and number density of the nanoparticles, while the platinum deposition current densities could be used to regulate the shape of the nanoparticles. In addition, the flower-like platinum nanoparticles showed high catalytic activity on electrochemical oxidation of methanol, and its activity was measured to be approximately 1.9 times that of bare platinum. PMID- 19217117 TI - An ATR-FTIR study of sulphate sorption on magnetite; rate of adsorption, surface speciation, and effect of calcium ions. AB - The adsorption of sulphate on magnetite was studied in-situ using ATR-FTIR spectroscopy. Synthetic magnetite particles were deposited on a ZnSe internal reflection element and the spectra of sulphate adsorbed at pH 4-8.5 were recorded. Two different ionic strengths were used viz. 0.01 M and 0.1 M NaCl. The spectra of adsorbed sulphate on magnetite coated ZnSe were compared with the spectra of sulphate solutions at the same pH values and in contact with uncoated ZnSe. The spectrum of adsorbed sulphate at pH 4 showed three maxima at 979, 1044, and 1115 cm(-1) indicating a monodentate adsorption in which the T(d) symmetry of SO(4)(2-) is lowered to C(3v). At pH 6.5, sulphate adsorbed as an outer-sphere complex with two weak bands appearing at 1102 and 980 cm(-1). Moreover, spectra of the adsorbed sulphate at pH 4 were recorded as a function of time and sulphate concentration. The equilibrium absorbance at different concentrations fitted a Langmuir type adsorption isotherm. The Langmuir affinity constant K at pH 4 was determined from the slope and intercept of the Langmuir plot to be K=1.2344x10(4) M(-1) and the Gibbs free energy of adsorption DeltaG(ads)(0) was estimated from this value to be -33.3 kJ/mol. Kinetic analysis indicated that adsorption at pH 4 is fast, whilst the desorption kinetic at the same pH is very slow. In addition, the effect of Ca ions on sulphate adsorption was also studied. It was shown that Ca ions increased the sulphate adsorption on magnetite at pH 8.5. PMID- 19217118 TI - Comments on "Adsorption of direct dyes from aqueous solutions by carbon nanotubes: determination of equilibrium, kinetics and thermodynamics parameters". AB - Two most suggested papers for pseudo-second-order kinetic model were published in 1984 and 1995 by Blanchard et al. and Ho, respectively. Blanchard et al. noted the overall exchange reaction of NH(+)(4) ions fixed in zeolite by divalent metallic ions in the solution using a second-order kinetic model. Ho used the pseudo-second-order kinetic model to the copper ion/peat adsorption system. The adsorption involved chemical bonding and cation exchange. In this comment citation error and quotation error were pointed. PMID- 19217119 TI - Synthesis of novel methacrylate based adsorbents and their sorptive properties towards p-nitrophenol from aqueous solutions. AB - The polymeric adsorbents were synthesized from 2-dimethylaminoethyl methacrylate (DMA) and [2-(methacryloyloxy)ethyl]dimethylhexadecylammonium bromide (DMAC(16)) monomers in the presence of ethylene glycol dimethacrylate (EDMA) cross-linking monomer using suspension polymerization technique and their adsorption efficiencies in the removal of p-nitrophenol from aqueous solutions were investigated. DMAC(16) monomer was prepared by means of modification of DMA monomer with 1-bromohexadecane. Adsorption experiments were carried out in a batch system and the experimental parameters were evaluated with respect to pH, agitation time, temperature and initial p-nitrophenol concentration. It was observed that the adsorbent poly[2 (methacryloyloxy)ethyl]dimethylhexadecylammonium bromide (p-DMAC(16)) prepared from DMAC(16) monomer was more effective in the removal of p-nitrophenol than the adsorbent poly(2-dimethylaminoethyl methacrylate) (p-DMA) prepared from DMA monomer. The effective pH ranges for the adsorption of p-nitrophenol onto p DMAC(16) and p-DMA were 2-12 and 3-9, respectively. Langmuir and Freundlich adsorption models were used to describe the isotherms and find isotherm constants. The Langmuir model was well agreed with experimental data for both adsorbents. The pseudo-first-order, pseudo-second-order, and intraparticle diffusion kinetic models were used to understand the mechanism of the adsorption process and it fitted very well the pseudo-second-order kinetic model for each adsorbent. Thermodynamic parameters such as activation energy and changes of free energy, enthalpy, and entropy were also evaluated for the adsorption of p nitrophenol onto each adsorbent. Additionally, reusability of the adsorbents was investigated and the results showed that both adsorbents can be employed many times without a significant loss in their adsorption capacities for the removal of p-nitrophenol from water. PMID- 19217120 TI - Electrochemically controlled release of alpha,beta,gamma,delta-tetrakis(4-N methylpyridyl)porphine from layer-by-layer thin films. AB - The release of alpha,beta,gamma,delta-tetrakis(4-N-methylpyridyl)porphyrin (TMPyP) from layer-by-layer assembled thin films composed of poly(allylamine hydrochloride) (PAH) and poly(acrylic acid) (PAA) was electrochemically controlled. The release rate of TMPyP was enhanced when a positive electrode potential (+1.2 to +1.5 V) was applied to the glassy carbon electrode on which TMPyP-loaded PAH/PAA film was coated, while the effect of lower electrode potentials (0 to +1.0 V) was negligibly small. The effect of applied potential on the release rate was more significant at pH 7.4 than in the solutions of pH 8.0 and 9.0. The electrochemically enhanced release of TMPyP was rationalized based on local pH change in the vicinity of the electrode surface, which was induced by electrolysis of H(2)O. PMID- 19217121 TI - Study of the interaction between a diblock polyelectrolyte PDMA-b-PAA and a gemini surfactant 12-6-12 in basic media. AB - The interactions between negatively charged diblock polyelectrolyte PDMA(71)-b PAA(59) and oppositely charged gemini surfactant hexylene-1,6 bis(dodecyldimethylammonium bromide) (12-6-12) in basic media were studied using dynamic light scattering, fluorescence spectroscopy, surface tension, and (1)H NMR. With increased addition of surfactant, the conformation of polyelectrolyte experienced changes from the initial unimer with open-extended PAA block, to the nano-scaled aggregates/complexes with a maximum hydrodynamic diameter (D(h)), and finally to the stable complexes with a smaller D(h). Accordingly, the value of D(h) during the whole process of increasing the surfactant concentration changed from 14-17 nm, to 184 nm, and to the final 70 nm, respectively. This transformation was driven by the electrostatic attractive/repulsive interactions, the hydrophobic interaction between hydrophobic surfactant tails, and the hydrophilicity of PDMA block. PMID- 19217122 TI - The susceptibility of young preschoolers to source similarity effects: confusing story or video events with reality. AB - This study examined children's tendency to confuse events that varied in source similarity, which was manipulated using different media of event presentation. In Experiment 1, children in two age groups (3- and 4-year-olds and 5- and 6-year olds) experienced a live presentation of an event, and another event was either heard from a story (low similarity group) or seen on a video (high similarity group). Immediately afterward, the children were asked to monitor the source of the events. The children in the low similarity group produced higher source discrimination scores than did the children in the high similarity group. Overall, the older children were better at source monitoring than were the younger children. In Experiment 2, the procedure was replicated except that the children's source monitoring was tested after a 4-day delay. When attributing the source of the story or video events, both 3- and 4-year-olds and 5- and 6-year olds in the low similarity group produced more accurate story or video attributions than did their age mates in the high similarity group. However, when attributing the source of the live events, only the 3- and 4-year-olds evidenced this effect of source similarity. The 5- and 6-year-olds in both the low and high similarity groups performed at ceiling levels for live discriminations. PMID- 19217123 TI - Characterization of surgical procedures in patients with mucopolysaccharidosis type I: findings from the MPS I Registry. AB - OBJECTIVE: To clarify the extent and chronology of surgical burden in relation to symptom onset and diagnosis in patients with mucopolysaccharidosis I (MPS I) as reported in the MPS I Registry, an international observational database. STUDY DESIGN: Analysis of surgical data from 544 patients enrolled in the MPS I Registry. Among all patients with at least 1 reported surgery, the number and frequency of procedures, and age at procedure, diagnosis, and symptom onset were collected overall, by patient, and by reported phenotype (Hurler, Hurler-Scheie, Scheie). RESULTS: Overall and by phenotype, approximately 75% of patients in the MPS I Registry reported at least 1 surgery. The most common were myringotomies and related procedures, hernia repair, adenoidectomy/tonsillectomy, and carpal tunnel release. Median age at first surgery was <5 years. A median of 3 to 4 surgeries was reported per patient. By age 1.5, 4, and 10 years, respectively, 22%, 44%, and 54% of patients reported > or = 2 surgeries. At least 1 surgery preceded diagnosis in 36%, 46%, and 63% of patients with Hurler, Hurler-Scheie, and Scheie, respectively. CONCLUSIONS: Pediatricians and pediatric surgeons need to be aware of the surgical burden of MPS I and be alert to its presenting signs and symptoms in children scheduled for surgery. PMID- 19217124 TI - Endothelial pulse amplitude testing: feasibility and reproducibility in adolescents. AB - OBJECTIVES: To test prospectively the reproducibility and feasibility of endothelial pulse amplitude testing (Endo-PAT), a novel Food and Drug Administration-approved technology, in healthy adolescents. STUDY DESIGN: We performed Endo-PAT testing on 2 different days separated by no more than 7 days in 30 healthy fasting adolescents, ages 13 to 19 years, to assess reproducibility and feasibility. The reported level of discomfort, as measured on a pain scale of 1 to 5, was documented. RESULTS: The mean difference in paired Endo-PAT indices was 0.12 (95% CI, -0.09-0.33; P = .24; intraclass correlation coefficient, 0.78), and the within-subject variation of Endo-PAT index was 0.16. The Endo-PAT index on test days 1 and 2 were 1.91 +/- 0.57 and 1.78 +/- 0.51 (mean plus or minus SD), respectively. All attempted studies (100%) were completed (95% CI, 88% 100%), and all completed studies (100%) could be analyzed (95% CI, 88%-100%). The median pain score was 1 on both days. CONCLUSION: In healthy adolescents, Endo PAT is feasible and has excellent reproducibility. This technology may provide an easy and reliable means of assessing endothelial function in the pediatric population. PMID- 19217125 TI - Seco-tabersonine alkaloids from Tabernaemontana corymbosa. AB - Two seco-tabersonine alkaloids, jerantiphyllines A and B, in addition to a tabersonine hydroxyindolenine, jerantinine H, and a recently reported vincamine alkaloid 7, were isolated from the leaf extract of the Malayan Tabernaemontana corymbosa and the structures were established using NMR and MS analysis. Biomimetic conversion of jerantinines A and E to their respective vincamine and 16-epivincamine derivatives were also carried out. PMID- 19217126 TI - A survey on canine leishmaniasis and phlebotomine sand flies in central Italy. AB - Zoonotic visceral leishmaniasis (ZVL) is a vector-transmitted zoonosis caused by the parasitic protozoan Leishmania infantum. Bloodsucking sand flies of the subfamily Phlebotominae are the obligatory insect hosts, and the dog is the only domestic reservoir. This study reports data from a survey of canine infection and sand fly phlebotomine monitoring in the province of Perugia in central Italy. The overall seroprevalence in a total of 100 dogs tested was 8% (95% confidence interval: 3.8-15.6%). Data analysis revealed that serological positivity was statistically associated with age (p-value=0.03) and the area where dogs lived. Standard blacklight traps employed for sampling Culicoides midges in bluetongue disease surveillance were used in phlebotomine monitoring. A total of 5698 sand flies were collected and the two species, Leishmania competent vectors, were identified, Phlebotomus perfiliewi (50%) and Phlebotomus perniciosus (30%). PMID- 19217127 TI - [The electroencephalogram in status epilepticus]. AB - Recordings of the electroencephalogram (EEG) play a major role in the management of patients with status epilepticus (SE). The EEG contributes to the diagnosis of SE, can be used to identify differential and syndromic diagnoses, and sometimes provides the etiologic diagnosis. EEG is helpful in monitoring therapeutic management and is an essential component of the follow up. The interpretation of the EEG in a patient with refractory SE is difficult, requiring clinical experience in this domain. We discuss the different modalities of EEG recording and their indications in emergency situations. PMID- 19217128 TI - [Sonography versus electrodiagnosis for the diagnosis of carpal tunnel syndrome in routine practice]. AB - INTRODUCTION: Many studies had been performed in the last years to prove the usefulness of ultrasonographic measurements of the median nerve in the diagnosis of carpal tunnel syndrome (CTS). We wanted to determine its reliability and to compare this technology with electromyography (EMG) in ordinary diagnostic conditions. METHODS: The study involved 90 wrists with suspected CTS, 35 controlateral wrists and 52 control wrists. The diagnosis of CTS was confirmed in 81 cases by the hand symptom diagram and the Tinnel and Phalen sign. The EMG examination evaluated medianulnar sensory latency difference to the ring finger and wrist-to-palm sensory conduction velocity. For the ultrasound diagnosis, the cross sectional area of the median nerve at the level of the pisiform bone, was considered. The sensitivity and specificity of the two techniques was calculated. RESULTS: Sensitive electroneurographic parameters showed a sensibility and specificity respectively of 79 and 80%. The cut-off point for ultrasound sensibility and specificity using ROC analysis was 11mm(2) for mean cross sectional area. Sensitivity and specificity found in this way were 72% and 56%. Reliability was good with intra- and inter-reader intraclass correlation coefficients of 0.99, and interobserver coefficient of 0.88. Sonography found seven CTS among the 17 clinical CTS with normal electrophysiological findings. A statistically correlation was found between the cross-sectional section and the sensitive electrophysiologic parameters (r=0.43, p<0.001). CONCLUSIONS: In our study, ultrasonographic diagnostic value are not as good as electrophysiological value, like found in recent literature, probably because of the composition of our group of patients which is including many causes of acroparesthesias. This can mean that in clinical practice, sonography is a complementary tool instead, for example in cases of equivocal EMG. PMID- 19217129 TI - [Metronidazole-related polyneuritis, convulsive seizures, and cerebellar syndrome. Contribution of MRI]. AB - INTRODUCTION: Neurological complications of metronidazole are rare, predominantly peripheral neuropathies, especially in patients on a long-term high-dose regimen. Cerebellar syndrome or seizures are less frequently reported. The concomitant occurrence of the three complications is exceptional. CASE REPORT: We report herein a case with these three complications as side effects of metronidazole. For the cerebellar syndrome, the T2-weighted brain MRI showed a rounded and well delimited zone of high signal intensity in the cerebellar dentate nuclei, extending up to the protuberance and the subthalamic nucleus, bilaterally and symmetrically. CONCLUSION: Neurological complications are possible when a treatment with metronidazole is prescribed for a long duration or at high dose. In our patient, the clinical abnormalities and MRI signs regressed a few months after treatment withdrawal. PMID- 19217130 TI - [Internet and amyotrophic lateral sclerosis treatment: what is wrong?]. PMID- 19217131 TI - Epidemiological study of hepatitis A, B and C in the largest Afro-Brazilian isolated community. AB - This study was conducted to estimate the prevalence and molecular epidemiological features of viral hepatitis A, B and C in the Kalunga population, which represents the largest Afro-Brazilian isolated community. Among 878 individuals studied, the overall prevalence of anti-hepatitis A virus antibodies was 80.9%, with a significant rise from 44.8% to near 100% between the first and fourth decade of life. Rates for hepatitis B surface antigen (HBsAg) and antibody to hepatitis B core antigen (anti-HBc) of 1.8% and 35.4%, respectively, were found. Increasing age, male gender, illiteracy and history of multiple sexual partners were associated with hepatitis B virus (HBV) infection. An occult HBV infection rate of 1.7% (5/295) was found among anti-HBc-positive individuals. HBV genotype A (subtype Aa) was dominant in this community. Only 5/878 individuals (0.6%) were positive for anti-hepatitis C virus (HCV). HCV RNA was detected in three of them, who were infected with genotype 1 (subtype 1a). These findings point out high, intermediate and low endemicity for hepatitis A, B and C, respectively, in the Kalunga community in Brazil. Circulation of HBV genotype A (subtype Aa) in this Afro-Brazilian isolated community indicates the introduction of this virus during the slave trade from Africa to Brazil. PMID- 19217132 TI - Clinical profile of 516 children affected by HIV in a tertiary care centre in northern India: 14 years of experience. AB - Increasing numbers of children affected by HIV are being recognised in northern India. The present study aimed to evaluate the clinical profile of 516 children affected by HIV at the Pediatric Allergy and Immunology Unit, Advanced Pediatric Center, Postgraduate Institute of Medical Education and Research, Chandigarh, India, during the period January 1994 to May 2008. In total, 454 children (327 boys and 127 girls) infected by HIV were analysed. The median age at presentation was 54 months. Of these children, 401 (88.3%) acquired the infection vertically and 26 (5.7%) acquired it through transfusion of blood/blood products. Moreover, 156 children (34.4%) were asymptomatic at presentation to hospital. Common clinical features included fever (36.6%), respiratory infections (31.7%), lymphadenopathy (30.0%), hepatosplenomegaly (21.8%) and diarrhoea (18.1%); 299 children (65.9%) were malnourished. Triple drug antiretroviral therapy was initiated in 205 children. Children receiving such therapy showed significant improvement in clinical and immunological parameters. Furthermore, follow-up rates improved markedly following free supply of the drug. Therapy was very well tolerated. To conclude, physicians looking after children need to be familiar with the varying clinical presentation of HIV infection. To the best of our knowledge, this is the largest paediatric series on HIV infection from a single centre from any developing country. PMID- 19217133 TI - Crystalline inclusions in hepatocytes and associated interhepatocytic macrophages from female Ohrid trout (Salmo letnica Kar.). AB - When characterizing the liver ultrastructure in Ohrid trout, we noticed that cells often displayed unusual cytoplasmic crystalline inclusions. Their morphology varied much, concerning the size, shape and electron density, showing aspects never reported in fish liver. In hepatocytes, the inclusions existed in close topographical relationship with dense bodies (often within them). Such "crystals" inclusions also existed in macrophages and in biliary passages lumina. Data revealed seasonal/breeding-related changes of the dense bodies and crystalline inclusions within hepatocytes; decreasing from the earliest to the more advanced ovary maturation stages. Additionally, based on the negative correlations between the amounts of dense bodies and of "crystals" and the ovary somatic ratio, we suggested there might be connections between the sex steroids status and the inclusions formation. A positive correlation between the quantities of "crystals" and of dense bodies further suggested that the inclusions might derive from normal processes, e.g., the females' cyclic liver changes, that involves dense bodies' turnover. However, a toxicological aetiology cannot be excluded. Additionally, multiple mechanisms can contribute to the "crystals" genesis. Facing literature reports and because the inclusions content seemed washed out during tissue processing, they are likely lipid in nature, but their exact composition and genesis require further analyses. PMID- 19217134 TI - Insights into the translational regulation of biologically active open reading frames of Pelargonium line pattern virus. AB - Pelargonium line pattern virus (PLPV), a proposed member of a prospective genus (Pelarspovirus) within family Tombusviridae, has a positive-sense, single stranded genomic RNA. According to previous predictions, it contains six open reading frames (ORFs) potentially encoding proteins of 27 (p27), 13 (p13), 87 (p87), 7 (p7), 6 (p6), and 37 kDa (p37). Using a variety of techniques we demonstrate that all predicted ORFs are functional, with the exception of (p13) and (p6). We also characterize a previously unidentified ORF which encodes a 9.7 kDa protein (p9.7) that is essential for viral movement. Furthermore, we present evidence that the single subgenomic RNA (sgRNA) produced by the virus directs synthesis of p7, p9.7 and p37. Remarkably, the translation of these totally unrelated proteins is coordinated via leaky-scanning. This mechanism seems to be favoured by the poor translation context of the start codon of ORF(p7), the non AUG weak initiation codon of ORF(p9.7) and the lack of additional AUG codons in any reading frame preceding ORF(p37). The results also suggest that precise regulation of protein production from the sgRNA is critical for virus viability. Altogether, the data supports the notion that PLPV belongs to a new genus of plant viruses. PMID- 19217135 TI - The microRNAs of Epstein-Barr Virus are expressed at dramatically differing levels among cell lines. AB - Epstein-Barr Virus (EBV) encodes multiple microRNAs (miRNAs) from two primary transcripts, BHRF1 and the BARTs. The expression of BHRF1 miRNAs is dependent on the type of viral latency, whereas the BART miRNAs are expressed in cells during all forms of latency. It is not known how these miRNAs are otherwise regulated, though. We have used quantitative, stem-loop, real-time PCR to measure the expression of EBV's miRNAs and found them to differ nearly 50- and 25-fold among all tested cell lines and among EBV-positive Burkitt's lymphomas, respectively. In addition, the expression of individual BART miRNAs within a cell can differ by 50-fold or more despite the fact these miRNAs are likely transcribed together as a single primary transcript. These measurements are illuminating: they indicate that few of EBV's miRNAs are expressed at levels comparable to those of cellular miRNAs in most cell lines and therefore likely function interdependently. PMID- 19217136 TI - Expression of the highly conserved vaccinia virus E6 protein is required for virion morphogenesis. AB - The vaccinia virus E6R gene (VACVWR062) is conserved in all members of the poxvirus family and encodes a protein associated with the mature virion. We confirmed this association and provided evidence for an internal location. An inducible mutant that conditionally expresses E6 was constructed. In the absence of inducer, plaque formation and virus production were severely inhibited in several cell lines, whereas some replication occurred in others. This difference could be due to variation in the stringency of repression, since we could not isolate a stable deletion mutant even in the more "permissive" cells. Under non permissive conditions, viral late proteins were synthesized but processing of core proteins was inefficient, indicative of an assembly block. Transmission electron microscopy of sections of cells infected with the mutant in the absence of inducer revealed morphogenetic defects with crescents and empty immature virions adjacent to dense inclusions of viroplasm. Mature virions were infrequent and cores appeared to have lucent centers. PMID- 19217137 TI - Long-term (1,243 days), low-temperature (4-15 degrees C), anaerobic biotreatment of acidified wastewaters: bioprocess performance and physiological characteristics. AB - The feasibility of long-term (>3 years), low-temperature (4-15 degrees C) and anaerobic bioreactor operation, for the treatment of acidified wastewater, was investigated. A hybrid, expanded granular sludge bed-anaerobic filter bioreactor was seeded with a mesophilic inoculum and employed for the mineralization of moderate-strength (3.75-10 kg chemical oxygen demand (COD)m(-3)) volatile fatty acid-based wastewaters at 4-15 degrees C. Bioprocess performance was assessed in terms of COD removal efficiency (CODRE), methane biogas concentration, and yield, and biomass retention. Batch specific methanogenic activity assays were performed to physiologically characterise reactor biomass. Despite transient disimprovements, CODRE and methane biogas concentrations exceeded 80% and 65%, respectively, at an applied organic loading rate (OLR) of 10 kgCODm(-3)d(-1) between 9.5 and 15 degrees C (sludge loading rate (SLR), 0.6 kgCOD kg[VSS](-1)d( 1)). Over 50% of the granular sludge bed was lost to disintegration during operation at 9.5 degrees C, warranting a reduction in the applied OLR to 3.75-5 kgCODm(-3)d(-1) (SLR, c. 0.4-0.5kgCOD kg[VSS](-1)d(-1)). From that point forward, remarkably stable and efficient performance was observed during operation at 4-10 degrees C, with respect to CODRE (>or=82%), methane biogas concentration (>70%) and methane yields (>4l(Methane)d(-1)), suggesting the adaptation of our mesophilic inoculum to psychrophilic operating conditions. Physiological activity assays indicated the development of psychroactive syntrophic and methanogenic populations, including the emergence of putatively psychrophilic propionate oxidising and hydrogenotrophic methanogenic activity. The data suggest that mesophilic inocula can physiologically adapt to sub-optimal operational temperatures: treatment efficiencies and sludge loading rates at 4 degrees C (day, 1243) were comparable to those achieved at 15 degrees C (day 0). Furthermore, long-term, low-temperature bioreactor operation may act as a selective enrichment for psychrophilic methanogenic activity from mesophilic inocula. The observed efficient and stable bioprocess performance highlights the potential for long-term, low-temperature bioreactor operation. PMID- 19217138 TI - In situ characterization of nitrifying biofilm: minimizing biomass loss and preserving perspective. AB - Methods for characterizing nitrifying bacteria within biofilms are of key importance to understand and optimize the nitrification kinetics of attached growth treatment facilities. In this work, we propose an analytical protocol based upon environmental scanning electron microscopy (ESEM) and confocal laser scanning microscopy (CSLM) in combination with fluorescent in situ hybridization (FISH) to characterize the structure of nitrifying biofilm as it remains attached to the original reactor substratum. This protocol minimizes the loss of mass and distortion of in situ perspective commonly associated with traditionally applied microscopic techniques and thereby enables a more accurate estimation of the nitrifying biomass within biofilm attached to the substratum. The use of ESEM eliminates the destructive preparatory procedures associated with traditional scanning electron microscopy and thus the loss of mass and shrinking of the samples. ESEM is used in this study to evaluate the percent coverage of the substratum with biofilm and the biofilm thickness. CLSM-FISH is used to determine cell counts in the biofilm and to characterize the undisturbed substratum/biofilm interface. By hybridizing and analyzing the nitrifying biofilm using CLSM as it remains attached to the substratum, the loss of material and distortion of in situ perspective associated with the biofilm detachment process is minimized. Moreover, by conducting the CLSM analysis directly on the nitrifying biofilm as it remains attached to the substratum it is shown that cell counts at the substratum/biofilm interface differ significantly from that located above the interface. PMID- 19217139 TI - Kinetics of hexavalent chromium removal from water by chitosan-Fe0 nanoparticles. AB - Chitosan-Fe(0) nanoparticles (chitosan-Fe(0)) were prepared using nontoxic and biodegradable chitosan as a stabilizer. Batch experiments were conducted to evaluate the influences of initial Cr(VI) concentration and other factors on Cr(VI) reduction on the surface of the chitosan-Fe(0). The overall disappearance of Cr(VI) may include both physical adsorption of Cr(VI) onto the chitosan-Fe(0) surface and subsequent reduction of Cr(VI) to Cr(III). The rate of reduction of Cr(VI) to Cr(III) can be expressed by a pseudo-first-order reaction kinetics. The rate constants increase with the increase in temperature and iron loading but decrease with the increase in initial Cr(VI) concentration and pH. The apparent activation energy is found to be 33 kJ mol(-1), which is characteristic of a chemically controlled reaction. Characterization with high-resolution X-ray photoelectron spectroscopy reveals that after the reaction, relative to Cr(VI) and Fe(0), Cr(III) and Fe(III) are the predominant species on the surface of chitosan-Fe(0). Chitosan has also been found to inhibit the formation of Fe(III) Cr(III) precipitation due to its high efficiency in chelating the Fe(III) ions. This study demonstrates that chitosan-Fe(0) has the potential to become an effective agent for in situ subsurface environment remediation. PMID- 19217140 TI - Polychlorinated dibenzo-p-dioxins, dibenzofurans, and polychlorinated biphenyls in semi-domesticated reindeer (Rangifer tarandus tarandus) and wild moose (Alces alces) meat in Finland. AB - Semi-domesticated reindeer and wild moose meat samples were analyzed for polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), and polychlorinated biphenyls (PCBs). Both calves and adults were studied. Individual reindeer and moose meat samples and pooled reindeer calf meat samples were collected from the northern, the middle, and the southern reindeer herding regions in Finland. Samples represented the edible parts of carcasses. In individual samples of reindeer the fat based WHO-PCDD/F-PCB-TEQ concentration was on average 3.2pgg(-1) in calves and 2.3pgg(-1) in adults. In moose calves the fat based WHO-PCDD/F-PCB-TEQ concentration (1.9pgg(-1)) was lower than in reindeer calves. WHO-PCDD/F-PCB-TEQ concentration in the adult moose samples was equal as in the adult reindeer samples. The mean fat based WHO-PCDD/F-PCB-TEQ concentration was highest in reindeer calf samples from the middle region. These samples contained also the highest content of fat. Individual samples of reindeer contained on average more WHO-PCB-TEQ than WHO-PCDD/F-TEQ, while the opposite was true for moose samples, and also samples of adult reindeer from the southern area. The contributions of PCDD/Fs and PCBs to the total TEQ were similar in the reindeer calves' pooled samples which were collected from more western regions than individual samples. PMID- 19217141 TI - 8-2 fluorotelomer alcohol aerobic soil biodegradation: pathways, metabolites, and metabolite yields. AB - The biodegradation pathways and metabolite yields of [3-(14)C] 8-2 fluorotelomer alcohol [8-2 FTOH, F(CF(2))(7)(14)CF(2)CH(2)CH(2)OH) in aerobic soils were investigated. Studies were conducted under closed (static) and continuous headspace air flow to assess differences in degradation rate and metabolite concentrations in soil and headspace. Aerobic degradation pathways in soils were in general similar to those in aerobic sludge and bacterial culture. (14)C mass balance was achieved in soils incubated for up to 7 months. Up to 35% (14)C dosed was irreversibly bound to soils and was only recoverable by soil combustion. The average PFOA yield was approximately 25%. Perfluorohexanoic acid (PFHxA) yield reached approximately 4%. (14)CO(2) yield was 6.8% under continuous air flow for 33 days. Three metabolites not previously identified in environmental samples were detected: 3-OH-7-3 acid [F(CF(2))(7)CHOHCH(2)COOH], 7-2 FT ketone [F(CF(2))(7)COCH(3)] and 2H-PFOA [F(CF(2))(6)CFHCOOH]. No perfluorononanoic acid (PFNA) was observed. The formation of 2H-PFOA, PFHxA, and (14)CO(2) shows that multiple -CF(2)- groups were removed from 8-2 FTOH. 7-3 Acid [F(CF(2))(7)CH(2)CH(2)COOH] reached a yield of 11% at day 7 and did not change thereafter. 7-3 Acid was incubated in aerobic soil and did not degrade to PFOA. 7 2 sFTOH [F(CF(2))(7)CH(OH)CH(3)], a transient metabolite, was incubated and degraded principally to PFOA. 7-3 Acid may be a unique metabolite from 8-2 FTOH biodegradation. The terminal ratio of PFOA to 7-3 acid ranged between 1.8-2.5 in soils and 0.6-3.2 in activated sludge, sediment, and mixed bacterial culture. This ratio may be useful in evaluating environmental samples to distinguish the potential contribution of 8-2 FTOH biodegradation to PFOA observed versus PFOA originating from other sources. PMID- 19217142 TI - Strategies to use phytoextraction in very acidic soil contaminated by heavy metals. AB - In microcosm experiments, the use of inorganic and organic amendments has been studied as potential agents to reduce heavy metal bioavailability in an acidic soil highly contaminated by Cu, Zn and Ni, that has to be remediated by phytoremediation. The concentrations of heavy metals in the original soil (O Soil) produced phytotoxic effects with a strong reduction in biomass yield that hinder the utilization of this technology. To overcome phytotoxicity the use of three immobilizing agents was evaluated. The results obtained showed that all the strategies decreased the mobile fractions of heavy metals in soil and increased the metal removal efficiency. In the case of Brassica juncea the best results for Zn and Ni were obtained after zeolites addition (Z-Soil) with an increase of about 6 times with respect to the value found in the O-Soil. In the case of Cu, the more efficient treatment was Ca(OH)(2) addition (Ca-Soil). The B. juncea plants accumulated Cu amounts 8 times greater than in the O-Soil. For this metal, relevant results were obtained also with compost, that increased the amount of Cu in the plants of 7 times with respect to the O-Soil. Similar results were obtained with Helianthus annuus the highest Zn and Ni accumulation was detected in the Z-Soil and compost-treated soils (C-Soil), with an increase of nearly 11 times with respect to the accumulation in the O-Soil. In the case of Cu the highest increase of total uptake was found in the C-Soil: 28 times higher than in the O-Soil. Total accumulation in Poa annua plants showed the highest removal efficiency in the Z-Soil for all metals. The values obtained increased of 4, 11 and 12 times for Cu, Zn and Ni, respectively. PMID- 19217143 TI - Mineralogical and geochemical controls of arsenic speciation and mobility under different redox conditions in soil, sediment and water at the Mokrsko-West gold deposit, Czech Republic. AB - Naturally contaminated soil, sediment and water at the Mokrsko-West gold deposit, Central Bohemia, have been studied in order to determine the processes that lead to release of As into water and to control its speciation under various redox conditions. In soils, As is bonded mainly to secondary arseniosiderite, pharmacosiderite and Fe oxyhydroxides and, rarely, to scorodite; in sediments, As is bonded mainly to Fe oxyhydroxides and rarely to arsenate minerals. The highest concentrations of dissolved As were found in groundwater (up to 1141 microg L( 1)), which mostly represented a redox transition zone where neither sulphide minerals nor Fe oxyhydroxide are stable. The main processes releasing dissolved As in this zone are attributed to the reductive dissolution of Fe oxyhydroxides and arsenate minerals, resulting in a substantial decrease in their amounts below the groundwater level. Some shallow subsurface environments with high organic matter contents were characterized by reducing conditions that indicated a relatively high amount of S(-2,0) in the solid phase and a lower dissolved As concentration (70-80 microg L(-1)) in the pore water. These findings are attributed to the formation of Fe(II) sulphides with the sorbed As. Under oxidizing conditions, surface waters were undersaturated with respect to arsenate minerals and this promoted the dissolution of secondary arsenates and increased the As concentrations in the water to characteristic values from 300 to 450 microg L(-1) in the stream and fishpond waters. The levels of dissolved As(III) often predominate over As(V) levels, both in groundwaters and in surface waters. The As(III)/As(V) ratio is closely related to the DOC concentration and this could support the assumption of a key role of microbial processes in transformations of aqueous As species as well as in the mobility of As. PMID- 19217144 TI - Natural attenuation and characterization of contaminants composition in landfill leachate under different disposing ages. AB - Chemical Oxygen Demand (COD) composition in landfill leachate would vary as the disposal time extended. Leachates with different ages were collected from Laogang Refuse Landfill of Shanghai, the largest landfill in China with a placement scale of 7600 t refuse per day. To characterize COD composition in leachate, samples were size-fractioned into suspended fractions (>0.45 microm), colloid fraction (0.45 microm70%) and in the proteins (55.0-79.6%) of the algal cells. Urea reduced the Ni content in the amino acid-carbohydrate but elevated its content in proteins, and shown significantly correlated with the protein content of the algal cells. Thus, nutrient enrichment could influence both metal uptake and its distribution in the subcellular structures and components of the phytoplankton, as well as its subsequent transfer in marine food chains. PMID- 19217195 TI - Immune modulation in the blue mussel Mytilus edulis exposed to North Sea produced water. AB - The discharge of oil well produced water (PW) provides a constant source of contaminants to the marine environment including polycyclic aromatic hydrocarbons, alkylated phenols, metals and production chemicals. High concentrations of PW cause adverse effects to exposed biota, including reduced survival, growth and reproduction. Here we explore the effects of PW on immune function in the blue mussel, Mytilus edulis. Mussels were exposed for 21 days to sublethal PW concentrations (0.125-0.5%) and cellular parameters were measured. Cell viability, phagocytosis and cytotoxicity were inhibited after exposure to 0.25% and 0.5% PW, whilst the 0.125% PW treatment produced significant increases in these biomarker responses. This biphasic response was only observed after 7 days exposure; longer exposure periods led to a reduction in immune parameters. Results indicate that PW concentrations close to the discharge point cause modulation to cellular immunity. The implications for longer-term disease resistance are discussed. PMID- 19217196 TI - Childhood sexual abuse and adult binge drinking among Kanak women in New Caledonia. AB - The long-term consequences of violence against women are poorly documented within the context of political domination, economic inequalities and rapid social change of indigenous communities. Using data from the first population study on violence against women and their consequences on health in New Caledonia, South Pacific, this article investigates the association between childhood sexual abuse and binge drinking among 441 adult Kanak women. Face-to-face standardised interviews were conducted in 2002-2003, among women aged 18-54 years drawn from the electoral rolls. Childhood sexual abuse before 15 years of age was reported by 11.6% of respondents. Nearly all the perpetrators (96%) were known to the victims (63% being a close relative). The rate of frequent binge drinking amongst the women within the last 12 months was 34%. After controlling for social and demographic factors, an independent association was found between childhood sexual abuse and current binge drinking. This study is the first to analyse the contribution of childhood sexual abuse to the likelihood of later heavy alcohol use in an indigenous population in the South Pacific. The findings call for improving and giving priority to care for children who are victims of violence to prevent long-term health consequences and to develop prevention programs aimed at alcohol-related behaviour in women, while taking into account simultaneous individual and collective factors. PMID- 19217197 TI - To what extent does socioeconomic status explain differences in health between Roma and non-Roma adolescents in Slovakia? AB - The Roma make up one of the largest ethnic groups in Europe. The few studies that are available report health among the Roma as considerably worse than that of the majority population, and virtually nothing is known about the health status of Roma adolescents. The purpose of this study was to compare the self-reported health outcomes of Roma adolescents living in Roma settlements with adolescents from the majority population and to assess the impact of socioeconomic status on the results obtained. We conducted a survey among Roma adolescents (N=330, mean age=14.5) and non-Roma adolescents (N=722, mean age=14.9) living in eastern Slovakia. We gathered data on sociodemographic position, self-rated health (using the SF-36), the occurrence of accidents and injuries during the past year, healthcare utilization during the past year, health complaints, mental health and social desirability. Roma adolescents reported poorer self-rated health, more accidents and injuries during the past year and more frequent use of healthcare during the past year, though fewer health complaints. Furthermore, they reported more prosocial behaviour than non-Roma. No differences appeared in total difficulties. Socioeconomic status decreased the association of ethnicity with health outcomes. Adjustment for social desirability had a significant effect on the differences for all outcomes, except for accidents and injuries during the past year. PMID- 19217198 TI - Bispectral index (BIS) helps predicting bad neurological outcome in comatose survivors after cardiac arrest and induced therapeutic hypothermia. AB - AIM OF THE STUDY: Determine the use of bispectral index (BIS) as prognostic tool in therapeutic hypothermia (TH) treated comatose survivors after cardiac arrest (CA), regardless of initial rhythm, location or cause. METHODS: Prospective, single-centre, unblinded, observational cohort study in an 18 bed general ICU in a tertiary teaching hospital. 45 consecutive comatose patients admitted after CA and treated with TH were included. All patients were sedated with a standardised protocol including neuromuscular blockade. Induced TH was started as soon as possible after arrival in the hospital and continued for 24h before slow rewarming. Sedation was stopped after reaching normothermia (36 degrees C). All patients benefited from maximal supportive intensive care and no therapeutic withdrawal or withholding was done unless bad neurological status was confirmed. Continuous BIS monitoring was performed over 72h in all patients. RESULTS: 14 patients presented BIS values of zero (0) during their ICU stay. At 6 months 11 patients were dead, 1 remained comatose and 2 had severe neurological sequelae (CPC3). No patient of this group had good neurological outcome or improved his neurological outcome between ICU and 6-month follow-up. 31 patients had BIS values higher than 0. At 6 months of those, 11 died, none remained comatose, 3 had bad neurological outcome (CPC3) and 17 had no or minor neurological sequelae (CPC1-2). Thus no correlation between good outcome and BIS values higher than 0 is possible. CONCLUSIONS: BIS values of 0 help predict bad neurological outcome after CA and induced hypothermia. PMID- 19217199 TI - Acute ischemic heart disease alters ventricular fibrillation waveform characteristics in out-of hospital cardiac arrest. AB - BACKGROUND: Although ventricular fibrillation waveform characteristics (VFWC) correlate with coronary perfusion pressure and may predict defibrillation outcome, recent animal data indicate that these waveform characteristics are altered in both acute myocardial infarction (AMI) and chronic coronary heart disease (CHD). We wanted to confirm these recent animal data in humans and explore the possibility for such characteristics to identify acute ischemia during cardiac arrest. METHODS: Data from all adult patients admitted to hospital after out-of-hospital VF cardiac arrest in Oslo between May 2003 and July 2007 were prospectively collected. Patients were categorized into one of four pre defined etiologic groups: patients with AMI (AMI only), patients with AMI and CHD (AMI and CHD), patients with previous CHD without evidence for a new AMI (CHD only), and patients with primary arrhythmia (PA). VFWC were analyzed from prehospital ECG tracings, and the different etiologic groups compared using ANOVA. RESULTS: One-hundred-and-one patients with ECG recordings usable for VF analysis could confidently be categorized; 16 with AMI only, 34 with AMI and CHD, 41 with CHD only and 10 with PA. The two VFWC median slope (MS) and amplitude spectral area (AMSA) were significantly depressed in patients with AMI only compared to both PA (MS p=0.008, AMSA p=0.035) and CHD only patients (MS p=0.008, AMSA p=0.006). CONCLUSIONS: AMI patients have depressed MS and AMSA compared to patients without AMI during VF cardiac arrest. VFWC might be helpful in identifying patients with AMI during cardiac arrest, but prospective clinical studies are warranted to assess its feasibility and clinical benefit. PMID- 19217200 TI - Early goal-directed hemodynamic optimization combined with therapeutic hypothermia in comatose survivors of out-of-hospital cardiac arrest. AB - BACKGROUND: Comatose survivors of out-of-hospital cardiac arrest (OHCA) have high in-hospital mortality due to a complex pathophysiology that includes cardiovascular dysfunction, inflammation, coagulopathy, brain injury and persistence of the precipitating pathology. Therapeutic hypothermia (TH) is the only intervention that has been shown to improve outcomes in this patient population. Due to the similarities between the post-cardiac arrest state and severe sepsis, it has been postulated that early goal-directed hemodyamic optimization (EGDHO) combined with TH would improve outcome of comatose cardiac arrest survivors. OBJECTIVE: We examined the feasibility of establishing an integrated post-cardiac arrest resuscitation (PCAR) algorithm combining TH and EGDHO within 6h of emergency department (ED) presentation. METHODS: In May, 2005 we began prospectively identifying comatose (Glasgow Motor Score<6) survivors of OHCA treated with our PCAR protocol. The PCAR patients were compared to matched historic controls from a cardiac arrest database maintained at our institution. RESULTS: Between May, 2005 and January, 2008, 18/20 (90%) eligible patients were enrolled in the PCAR protocol. They were compared to historic controls from 2001 to 2005, during which time 18 patients met inclusion criteria for the PCAR protocol. Mean time from initiation of TH to target temperature (33 degrees C) was 2.8h (range 0.8-23.2; SD=h); 78% (14/18) had interventions based upon EGDHO parameters; 72% (13/18) of patients achieved their EGDHO goals within 6h of return of spontaneous circulation (ROSC). Mortality for historic controls who qualified for the PCAR protocol was 78% (14/18); mortality for those treated with the PCAR protocol was 50% (9/18) (p=0.15). CONCLUSIONS: In patients with ROSC after OHCA, EGDHO and TH can be implemented simultaneously. PMID- 19217202 TI - The multifunctionality of agriculture and contractual policies. A comparative analysis of France and The Netherlands. AB - The process of CAP reform has reorientated the development of agriculture, more towards the principles of rural development and agricultural multifunctionality. These transformations have been accompanied by a shift in the principles and tools used by policy-makers, necessary to implement these new orientations. The emergent contractual policies represent a renewal of public intervention in agriculture. This article explores the implementation of these policy instruments in two countries, France and the Netherlands. The content and implementation of these policies differs greatly between the two countries, reflecting their different conceptions of multifunctionality. The Netherlands limited the scope of the policy instrument to nature and landscape protection, while France adopted a wider definition of multifunctionality. These differences are partly explained by the particularities of the national context to which those policies have been applied. France is characterized by an institutionalized system of co-management, in which farmers' organizations work closely and have a privileged relationship with the state. Although this mode of governance is weakening and becoming vulnerable, it proved to be highly effective during the 1990s and contributed to the adoption of a broad vision of multifunctionality. By contrast, the selective definition of multifunctionality within Dutch policy is partly due to the co evolution of strong environmentalist sympathies, together with a liberalization of agricultural management. These divergences are perceptible in the policies that were adopted as well as in the configurations of power that were at stake during the policy-making processes. PMID- 19217201 TI - Valproic acid (VPA), a histone deacetylase (HDAC) inhibitor, diminishes lymphoproliferation in the Fas -deficient MRL/lpr(-/-) murine model of autoimmune lymphoproliferative syndrome (ALPS). AB - OBJECTIVE: Autoimmune lymphoproliferative syndrome (ALPS) is a disorder of apoptosis, often presenting in childhood. Similarly, MRL/lpr(-/-) mice homozygous for Fas mutations develop an ALPS-like disease with autoimmunity, lymphadenopathy, splenomegaly, and expansion of double-negative T cells. Currently, there are no proven therapies with adequate safety margins for sustained abolition of the lymphoproliferation associated with ALPS. We sought to test the ability of valproic acid (VPA), a histone deacetylase inhibitor, to induce apoptosis and inhibit lymphoproliferation. MATERIALS AND METHODS: Human peripheral blood mononuclear cells from patients with ALPS and normal controls were tested in vitro to determine the efficacy of VPA at inducing cell death. VPA was used in vivo to control lymphoproliferation in MRL/lpr(-/-) mice, a model for ALPS. RESULTS: VPA induced cell death in vitro, and was partially inhibited by the pan caspase inhibitor, Z-VAD-FMK. MRL/lpr(-/-) mice treated with VPA for 8 weeks showed significant reductions in spleen and lymph node weights and cellularity compared to controls. A concomitant decrease in double-negative T cells was observed in the spleen, lymph nodes, and peripheral blood. Serum levels of VPA peaked 1 hour after injection, and a 2.5-fold increase in histone acetylation was observed in the spleen at 4 hours after injection. CONCLUSION: Based on our data, VPA is effective at reducing lymphoproliferation in mice, and is currently being studied in a clinical trial as a lympholytic agent in patients with ALPS. PMID- 19217203 TI - Bilateral basal ganglia and unilateral cortical involvement in a diabetic uremic patient. AB - We report a 57-year-old woman with uremic encephalopathy who presented with dysarthria, dysphagia, hypophonia, and drowsiness. The patient's radiologic findings were rather unusual in that magnetic resonance imaging (MRI) showed abnormal findings involving the basal ganglia bilaterally and frontal cortex unilaterally. After intensified hemodialysis, her symptoms and follow-up brain MRI showed marked improvement. We postulated that the underlying mechanism of uremic encephalopathy based on diffusion-weighted imaging and apparent diffusion coefficient maps. PMID- 19217204 TI - Targeting the RAF-MEK-ERK pathway in cancer therapy. AB - The clinical success of selective kinase inhibitors, such as imatinib and erlotinib, as therapeutic agents for several human cancers has prompted substantial interest in the further development and clinical testing of such inhibitors for a wide variety of malignancies. While much of this effort has been focused on the receptor tyrosine kinases, including EGFR, HER2, PDGF receptor, c KIT, and MET, inhibitors of serine/threonine kinases are also beginning to emerge within discovery pipelines. Among these kinases, the RAF and MEK kinases have received substantial attention, owing largely to the relatively high frequency of activating mutations of RAS ( approximately 20% of all human cancers), an upstream activator of the well established RAF-MEK-ERK signaling cascade, as well as frequent activating mutations in the BRAF kinase ( approximately 7% of all human cancers). Here, we summarize the current state of development of kinase inhibitors directed at this signaling pathway, a few of which have already demonstrating favorable toxicity profiles as well as promising activity in early phase clinical studies. PMID- 19217205 TI - Epidermal growth factor receptor vIII enhances tumorigenicity and resistance to 5 fluorouracil in human hepatocellular carcinoma. AB - We investigated whether EGFRvIII contributes to tumorigenicity and resistance to 5-FU in HCC cell lines. Our results show that several HCC cell lines have EGFRvIII expression. EGFRvIII-positive HCC cells grew more rapidly and had a lower sensitivity to 5-FU than EGFRvIII-negative HCC cells. For further analysis of the biological characteristics of EGFRvIII, an EGFRvIII or EGFR expression cassette was introduced into the HCC cell line, Huh-7. Compared with Huh-7 cells and Huh7-EGFR cells, Huh7-EGFRvIII not only exhibit significantly increase of cell growth in vitro and in vivo but also show enhanced migration in vitro. Furthermore, 5-FU has significantly lower inhibition effect on Huh7-EGFRvIII cells then on both Huh-7 and Huh7-EGFR cells in vitro and in vivo. Collectively, these results demonstrate that EGFRvIII plays a pivotal role in tumorigenicity and enhanced 5-FU resistance of HCC. PMID- 19217206 TI - v-Src-mediated transformation suppresses the expression of focal adhesion protein vinexin. AB - Expression of focal adhesion protein vinexin is reported to be altered in several cancer tissues; however, the mechanism of expressional change in vinexin is not known. Here we report the suppression of vinexin expression according to cellular transformation by v-Src. We found that vinexin expression was down-regulated both at the mRNA level and at the post-transcriptional level in v-Src-transformed cells. Both mTOR and MEK/ERK signals were involved in the suppression. Inhibition of these pathways by pharmacological treatment partially restored both vinexin protein and mRNA expression. Moreover, re-expression of vinexin in v-Src transformed cells suppressed cell migration. Taken together, these observations suggest that cellular transformation by v-Src suppressed vinexin expression and that down-regulation of vinexin may be associated with oncogenic transformation. PMID- 19217207 TI - TRAP1, a novel mitochondrial chaperone responsible for multi-drug resistance and protection from apoptotis in human colorectal carcinoma cells. AB - TRAP1 is a component of a pro-survival mitochondrial pathway up-regulated in tumor cells. The evaluation of TRAP1 expression in 26 human colorectal carcinomas showed up-regulation in 17/26 tumors. Accordingly, TRAP1 levels were increased in HT-29 colorectal carcinoma cells resistant to 5-fluorouracil, oxaliplatin and irinotecan. Thus, we investigated the role of TRAP1 in multi-drug resistance in human colorectal cancer. Interestingly, TRAP1 overexpression leads to 5 fluorouracil-, oxaliplatin- and irinotecan-resistant phenotypes in different neoplastic cells. Conversely, the inhibition of TRAP1 activity by TRAP1 ATPase antagonist, shepherdin, increased the sensitivity to oxaliplatin and irinotecan in colorectal carcinoma cells resistant to the single agents. These results suggest that the increased expression of TRAP1 could be part of a pro-survival pathway responsible for multi-drug resistance. PMID- 19217208 TI - Physical activity and low back pain: a U-shaped relation? AB - Being physically active is often suggested to be important in the prevention and management of low back pain. This simple view does not take into account that the relation between the level of activity and back pain may be a U-shaped curve - i.e. both inactivity and excessive activities (back-unhealthy activity) present an increased risk for back pain. We explored the U-shaped association between physical activity and chronic low back pain (3 months duration) by analyzing cross-sectional data from the Dutch population-based Musculoskeletal Complaints and Consequences Cohort study (DMC(3), 1998) of a sex-age stratified sample of 25 years and older (n=3364). Type of activity (daily routine, leisure time and sport activity), intensity of and time spent on these activities, and back exertion of sport activities were taken into account. Physical activity was not associated with chronic low back pain (CLBP) when studied by the dimension of activity, by the intensity or by the duration of physical activity. Only engaging in sport activity was associated with less CLBP (OR 0.78: 95% CI 0.66-0.93). The extremes of the total physical activity pattern were associated with CLBP. A moderate increased risk for CLBP was found for both participants with a sedentary lifestyle (OR 1.31: 95% CI 1.08-1.58) and for those being involved in physical strenuous activities (OR 1.22: 95% CI 1.00-1.49). This was especially true for women (sedentary: OR 1.44: 95% CI 1.10-1.83; physically active: OR 1.36: 95% CI 1.04-1.78). This study provides some evidence that the relation between physical activity and CLBP is U-shaped. PMID- 19217209 TI - Involvement of voltage-gated sodium channels blockade in the analgesic effects of orphenadrine. AB - Orphenadrine is a drug acting on multiple targets, including muscarinic, histaminic, and NMDA receptors. It is used in the treatment of Parkinson's disease and in musculoskeletal disorders. It is also used as an analgesic, although its mechanism of action is still unknown. Both physiological and pharmacological results have demonstrated a critical role for voltage-gated sodium channels in many types of chronic pain syndromes. We tested the hypothesis that orphenadrine may block voltage-gated sodium channels. By using patch-clamp experiments, we evaluated the effects of the drug on whole-cell sodium currents in HEK293 cells expressing the skeletal muscle (Nav1.4), cardiac (Nav1.5) and neuronal (Nav1.1 and Nav1.7) subtypes of human sodium channels, as well as on whole-cell tetrodotoxin (TTX)-resistant sodium currents likely conducted by Nav1.8 and Nav1.9 channel subtypes in primary culture of rat DRG sensory neurons. The results indicate that orphenadrine inhibits sodium channels in a concentration-, voltage- and frequency-dependent manner. By using site-directed mutagenesis, we further show that orphenadrine binds to the same receptor as the local anesthetics. Orphenadrine affinities for resting and inactivated sodium channels were higher compared to those of known sodium channels blockers, such as mexiletine and flecainide. Low, clinically relevant orphenadrine concentration produces a significant block of Nav1.7, Nav1.8, and Nav1.9 channels, which are critical for experiencing pain sensations, indicating a role for sodium channel blockade in the clinical efficacy of orphenadrine as analgesic compound. On the other hand, block of Nav1.1 and Nav1.5 may contribute to the proconvulsive and proarrhythmic adverse reactions, especially observed during overdose. PMID- 19217210 TI - Associations between nematode larval challenge and gastrointestinal tract size that affect carcass productivity in sheep. AB - Effects of gastrointestinal parasitism on sheep productivity are usually described using live weight change, however carcass productivity is more accurately described using dressing percentage (carcass weight as a proportion of live weight). This experiment had a 2x2x2 factorial design whereby 10-month-old Merino wethers were fed lucerne (Medicago sativa) diets (fresh lucerne or lucerne chaff) with 2 levels of carboxymethycellulose (CMC) inclusion (0% or 8% CMC) and nematode larval challenge (no larval challenge or 10,000 Teladorsagia circumcincta and 10,000 Trichostrongylus colubriformis per week). Sheep were weighed and euthanased 50 or 51 days after larval challenge and CMC supplementation commenced. Weight of the carcass (hot standard carcass weight) and gastrointestinal organs (full and empty) were recorded and expressed as a proportion of live weight. Larval challenged sheep had a worm egg count (mean+/ standard error) of 173+/-38 eggs per gram of faeces and total worm count of 30,237+/-2013 at slaughter. Larval challenged sheep had 1.3% lower dressing percentage (p=0.048), and 2% heavier full (p=0.007) and 1.2% heavier empty gastrointestinal tracts (p=0.012) compared to unchallenged sheep. There was no effect of CMC inclusion or lucerne type (fresh or chaff) on gastrointestinal tract weight or dressing percentage. Larval challenged sheep had 1.1% heavier full (p<0.001) and 0.6% heavier empty (p<0.001) small intestines, and 0.6% heavier full (p=0.005) and 0.3% heavier empty (p=0.026) large intestines compared to unchallenged sheep. Use of live weight change or other measures based on live weight (e.g. feed conversion efficiency) to assess the impact of nematode challenge in sheep may underestimate carcass productivity losses associated with larval challenge in sheep even at moderate levels of larval intake and without overt clinical signs of parasitism. Measurement of carcass weight and/or lean meat yield may better reflect the true economic effects of parasitism in sheep. PMID- 19217211 TI - Trichinella britovi and Trichinella spiralis mixed infection in a horse from Poland. AB - Trichinella infections in horses continue to represent a health problem and, despite the rarity of infection, it is necessary to continue to control properly horse meat. In 2008, a 10-year-old horse imported from Poland to Italy for consumption found to have been positive at the digestion test. Both Trichinella britovi and Trichinella spiralis larvae in a proportion of 4:1 were detected in the horse muscles. This is the first report of a mixed Trichinella species infection in a horse. The epidemiological investigation revealed that the infected horse originated from a small farm about 120km from Warsaw and the horse owner had bought the horse at a horse market. The findings suggest that the horse was fed more than once with infected meat. PMID- 19217212 TI - The influence of attachment on perceived stress and cortisol response to acute stress in women sexually abused in childhood or adolescence. AB - The long-term implications of sexual abuse in childhood or adolescence (CSA) have been relatively well documented regarding attachment (disorganized attachment in childhood, unresolved trauma in adulthood), stress reactions (altered patterns of stress reactivity under experimental conditions), and psychopathology. Attachment has been shown to mediate the implications of CSA, namely on psychopathology. The implication of attachment on stress responses of abused persons has not been documented. Twenty-seven 20-46 years old women who had experienced episodes of CSA, and 17 controls have been interviewed using the Adult Attachment Interview. Sixty-three percent of abused women presented an unresolved trauma (12% for the controls). Thirty-six women (14 controls and 22 abused) came again to the laboratory for a session involving an experimental stress challenge (TSST). Subjects provided repeated appreciations of perceived stress on visual analogue scales and saliva samples were collected to assay cortisol levels. Whereas abused women with unresolved trauma showed the highest levels of perceived stress, they simultaneously presented the most suppressed cortisol reactions (there were significant post hoc differences between "unresolved abused" and controls on the increase of perceived stress and on cortisol recovery after the acute stress). It is suggested that important stressful experiences (such as CSA), especially when they have not been psychologically assimilated, may cause a disconnection, during subsequent mildly stressful circumstances, between the perception of stress and natural defensive body reactions. PMID- 19217213 TI - Poor vitamin D status may contribute to high risk for insulin resistance, obesity, and cardiovascular disease in Asian Indians. AB - Asian Indians are highly prone to insulin resistance syndrome, obesity, diabetes, and coronary disease. At any given BMI, they tend to have more body fat and more central fat than other groups - yet their insulin resistance is disproportionately high relative to their body composition. They are also tend to have very poor vitamin D status, even in UV-drenched India, primarily owing to highly pigmented skin and a cultural tendency to avoid direct sun exposure. The resulting up-regulation of parathyroid hormone (PTH) arguably may play a role in their high risk for insulin resistance and associated pathologies. There is suggestive evidence that moderate elevations of PTH may promote insulin resistance, weight gain, hypertension, left ventricular hypertrophy, and the acute phase response, while increasing risk for ischemic arrhythmias and cardiovascular mortality. Controlled studies should assess the impact of optimal vitamin D supplementation, with or without added calcium, on risk factors associated with insulin resistance in Asian Indians, as well as in other highly pigmented urbanized ethnic groups that are at high risk for insulin resistance and obesity. PMID- 19217214 TI - Can uncompensated higher order aberration profile, or aberropia be responsible for subnormal best corrected vision and pseudo-amblyopia. AB - BACKGROUND: Higher order aberrations (HOA) are components of wavefront distortion which cannot be corrected by conventional spectacle correction. Normally they constitute up to 15% of wavefront distortion in the ocular media. The HOA interact in a nonlinear method and tend to compensate each other in most conditions. HYPOTHESES: We hypothesize that abnormally increased and uncompensated higher order aberration profile in certain cases may limit the amount of best corrected visual acuity to subnormal levels and produce a pseudo ambylopia like picture. We term this entity as aberropia. EVALUATION OF THE HYPOTHESIS: Higher order aberrations (HOA) have been proven to have supporting role in visual function in association with lower order aberrations. Normally HOA tend to compensate for each other and have an effect much less than what would be expected if they would have acted separately. This unique state of compensation may be lost in certain cases, for example, in patients who have undergone laser refractive surgery without consideration for HOA correction or in partial correction of a pathological eye like severe keratoconus. Occasionally investigators have noticed that patients with high induced aberrations after refractive surgery cannot achieve spectacle correction to get a 20/20 vision. Conversely, there is evidence for gain of vision after wavefront guided refractive surgery and phakic intraocular lens implantations in some cases which cannot be explained only by retinal image change. CHALLENGES TO CONCLUSIVELY PROVE THE HYPOTHESES: With the current level of advancement in refractive surgery, we can correct lower order aberration to a near zero level. However it is not as easy to do the same with higher order aberrations because of variability in assessment, evolving laser beam profiles and factors associated with the surgery itself and wound healing. All the four can alter higher order aberrations. IMPLICATIONS OF THE HYPOTHESES: Understanding of uncompensated higher order profiles leading to loss of BCVA, or aberropia, would be a paradigm shift in understanding of the role of higher order aberrations on visual function. With improvement in diagnostic and treatment methods, these patients with a subnormal vision, may be treated to achieve their maximum visual potential. PMID- 19217215 TI - Long-term therapy with proton pump inhibitors is associated with decreased bone density. PMID- 19217216 TI - Transporter associated with antigen processing and the chaperone tapasin: are non classical HLA genes keys to the pathogenesis of schizophrenia? AB - Schizophrenia is a mind-destructive disease. Family and twin studies have indicated an equal contribution of genetic endowment and environmental factors in the pathogenesis of schizophrenia. Recently Chlamydiaceae species has been identified as a major factor in the pathogenesis of schizophrenia, suggesting defective immune responses of schizophrenic patients against this environmental factor. Immune responses against Chlamydiaceae species are controlled by immunogenetic factors. Successful responses against microbes depend on the presentation of immunogenic peptides by HLA molecules, which are encoded by a highly polymorphic gene system. Until now several HLA alleles or HLA antigens have been found associated with schizophrenia by some researchers but not by others. This could be explained by failing immune responses against different microbes or different immune responses against the same microbe. Another explanation, not contradictory rather supplementary, is the participation of non classical HLA genes in the immune response and thus in the disease development. Variants of these genes, i.e. alleles, which control transportation and loading of microbial peptides onto HLA molecules, could prevent clearing of immune cell infection by selection of non-immunogenic peptides for HLA presentation. To generate support for our hypothesis we determined in a small group of schizophrenic patients and control individuals allele frequencies of the transporter proteins TAP1/TAP2, which select the immunoproteasome-tailored peptides for transportation. We determined also frequencies of TAPASIN alleles, which encode chaperons and also may select peptides for loading on MHC molecules. Our pilot study included 20 patients and 162 control individuals. We found significant associations between schizophrenia and TAP1 allele frequencies (P=9.95x10(-3), chi(2)=12.36) as well as TAPASIN allele frequencies (P=2.8x10( 2), chi(2)=5.3). This suggests that variants of these two genetic systems could influence the disease process of schizophrenia. Furthermore, these genes belong to the family of ABC transporter proteins and may also influence the efficiency of drugs and thus of therapeutic modalities. Our investigations require certainly larger patient panels to prove our hypothesis and our results to be correct. PMID- 19217217 TI - Dysadherin expression in gastrointestinal stromal tumors (GISTs). AB - The purpose of this study was to detect the protein and mRNA expression of dysadherin and to investigate the clinical significance of dysadherin expression in gastrointestinal stromal tumors (GISTs). Twenty fresh GISTs samples were used for testing the mRNA of dysadherin and E-cadherin by reverse-transcription polymerase chain reaction (RT-PCR). In addition, 54 paraffin blocks of GISTs cases were also used for the investigation of dysadherin and E-cadherin using immunohistochemistry. The dysadherin expression level was significantly correlated with GISTs risk stratification (chi(2)=5.769, P<0.05), but was not related to age, sex, histologic subtype, and location (P>0.05). There was only faint or absent E-cadherin protein expression in GISTs. The expression level of dysadherin is related to the risk of GISTs. Dysadherin upregulation, which may cause loss of E-cadherin, is one of the reasons for GISTs recurrence and metastasis. It seems that dysadherin protein detection is a promising method for GISTs prognostication. PMID- 19217218 TI - Hepatocellular carcinoma with massive lymphoid infiltration: a regressing phenomenon? AB - Spontaneous regression of hepatocellular carcinoma (HCC) is extremely rare. Among the numerous proposed mechanisms of spontaneous regression for HCC, immunological factors, which have yet to be fully understood, may have the most important roles in this rare phenomenon. A regressing HCC with lymphoid stroma is reported. A hepatic mass was detected in a 57-year-old man by abdominal computed tomography during a follow-up health check. The resected tumor was 2.7 x 2.4cm(2) in size, and was composed of two distinct nodules with a complete rim of fibrous tissue separating the tumor from the adjacent liver parenchyma. Microscopically, one nodule was replaced by a remarkable infiltration of inflammatory cells with insignificant amounts of vital tumor cells. The other nodule demonstrated a small portion of well- to moderately-differentiated hepatocellular carcinoma cells forming 2-3 thick trabeculae with massive inflammatory infiltrates. Infiltrating inflammatory cells were composed of T cells (CD4+>CD8+), macrophages, B cells, and other cells in order of prevalence. Epstein-Barr virus was not detected. After 60 months of follow-up, the patient is in good health without evidence of tumor recurrence. PMID- 19217219 TI - Stereotactic radiotherapy of intracranial tumors: a comparison of intensity modulated radiotherapy and dynamic conformal arc. AB - PURPOSE: Intensity-modulated radiotherapy (IMRT) and dynamic conformal arc (DCA) are two state-of-the-art techniques for linac-based stereotactic radiotherapy (SRT) using the micromultileaf collimator. The purpose of this planning study is to examine the relative merits of these techniques in the treatment of intracranial tumors. MATERIALS AND METHODS: SRT treatment plans were made for 25 patients with a glioma or meningioma. For all patients, we made an IMRT and a DCA plan. Plans were evaluated using: target coverage, conformity index (CI), homogeneity index (HI), doses in critical structures, number of monitor units needed, and equivalent uniform dose (EUD) in planning target volume (PTV) and critical structures. RESULTS: In the overall comparison of both techniques, we found adequate target coverage in all cases; a better mean CI with IMRT in concave tumors (p = 0.027); a better mean HI with DCA in meningiomas, complex tumors, and small (< 92 mL) tumors (p = 0.000, p = 0.005, and p = 0.005, respectively); and a higher EUD in the PTV with DCA in convex tumors (gliomas) and large tumors (p = 0.000 and p = 0.003, respectively). In all patients, significantly more monitor units were needed with IMRT. The results of the overall comparison did not enable us to predict the preference for one of the techniques in individual patients. The DCA plan was acceptable in 23 patients and the IMRT plan in 19 patients. DCA was preferred in 18 of 25 patients. CONCLUSIONS: DCA is our preferred SRT technique for most intracranial tumors. Tumor type, size, or shape do not predict a preference for DCA or IMRT. PMID- 19217220 TI - An isogenic Actinobacillus pleuropneumoniae AasP mutant exhibits altered biofilm formation but retains virulence. AB - AasP, an autotransporter serine protease of Actinobacillus pleuropneumoniae, has been shown to be expressed in necrotic porcine lung tissue. Based on the hypothesis that AasP might play an important role in A. pleuropneumoniae adhesion and virulence by processing other surface-associated proteins, the predicted catalytic site of AasP was deleted and the isogenic mutant, AP76DeltaaasP, was compared to the wild-type strain in a biofilm assay as well as an aerosol infection model. AP76DeltaaasP showed increased adherence compared to the wild type strain under standard culturing conditions as well as under NAD restriction. No significant differences between AP76 wild-type and AP76DeltaaasP were observed upon experimental infection of pigs, indicating that AasP does not play a crucial role in A. pleuropneumoniae virulence. PMID- 19217221 TI - Genotyping of Chlamydophila abortus strains by multilocus VNTR analysis. AB - Chlamydophila (C.) abortus is the causative agent of ovine enzootic abortion with zoonotic potential whose epidemiology has been held back because of the obligate intracellular habitat of the bacterium. In the present study, we report on a molecular typing method termed multiple loci variable number of tandem repeats (VNTR) Analysis (MLVA) for exploring the diversity of C. abortus. An initial analysis performed with 34 selected genetic loci on 34 ruminant strains including the variant Greek strains LLG and POS resulted in the identification of five polymorphic loci, confirming the widely held notion that C. abortus is a very homogeneous species. Analysis of additional 111 samples with the selected five loci resulted in the classification of all strains into six genotypes with distinct molecular patterns termed genotypes [1] through [6]. Interestingly, the classification of the isolates in the six genotypes was partly related to their geographical origin. Direct examination of clinical samples proved the MLVA to be suitable for direct typing. Analysis of the genomic sequences in six C. abortus prototypes of amplicons generated with each of the five selected VNTR primers revealed that variation between genotypes was caused by the presence or absence of coding tandem repeats in three loci. Amplification of Chlamydophila psittaci reference strains with the five selected VNTR primers and of the six C. abortus prototype strains with the eight VNTR primers established for the typing of C. psittaci [Laroucau, K., Thierry, S., Vorimore, F., Blanco, K., Kaleta, E., Hoop, R., Magnino, S., Vanrompay, D., Sachse, K., Myers, G.S., Bavoil, P.M., Vergnaud, G., Pourcel, C., 2008. High resolution typing of Chlamydophila psittaci by multilocus VNTR analysis (MLVA). Infect. Genet. Evol. 8(2), 171-181] showed that both MLVA typing systems were species-specific when all respective VNTR primer sets were used. In conclusion, the newly developed MLVA system provides a highly sensitive, high-resolution and easy-to-perform tool for the differentiation of C. abortus isolates of different origin, which is suitable for molecular epidemiological studies. PMID- 19217222 TI - Immune response against equine gammaherpesvirus in Icelandic horses. AB - Horses are hosts to two types of gammaherpesviruses, equine herpes virus (EHV) 2 and 5. While EHV-2 is ubiquitous in adult horses, EHV-5 has been less frequently described. Due to strong serological cross-reactivity, EHV-2 and -5 cannot be discriminated in broad spectrum antibody tests and are thus commonly referred to as gamma-EHV. Total IgG and IgG subclass response against gamma-EHV were determined in serum from 41 healthy Icelandic horses, thereof 20 adults, 10 foals aged 10 months, and 11 foals aged 1-4 months. Additionally, in 10 of the adult horses, interferon (IFN)-gamma and interleukin (IL)-4 expression were measured by real-time PCR in white blood cells upon in vitro stimulation with EHV-2. With the exception of one orphan foal, all tested individuals were seropositive for gamma EHV. All but one adult had high titer of EHV-specific IgG4/7 (IgGb) in combination with much lower titer of IgG1 (IgGa) and IgG3/5 (IgG(T)), indicating a stabilized response. IgG titer and subclasses in the foals showed considerably more variation, possibly dependant on maternal antibodies and/or recent infection. In all the 10 horses tested for cytokine expression, IFN-gamma production exceeds production of IL-4. These results indicate that equine gammaherpesvirus infection is characterized by an induction of IgG1, IgG4/7 and IgG3/5 with prevailing IgG4/7 and cytokine profile dominated by IFN-gamma. To our knowledge, this is the first report on the cytokine and IgG subclass response against gamma-EHV in horses. PMID- 19217223 TI - Influence of ventilation mode on neonatal cerebral blood flow and volume. AB - BACKGROUND: Cerebral hemodynamics is supposed to be influenced by the different ventilation approach. Ventilation support can be classified as non-invasive (N CPAP) or invasive (SIMV and HFV), the last known to induce endotrauma. Our aim was the non-invasive NIRS assessment of neonatal absolute cerebral blood flow (CBF) and relative cerebral blood volume changes (DeltaCBV) during synchronized intermittent ventilation (SIMV), or high frequency ventilation (HFV) and nasal continuous positive airways pressure (CPAP). METHODS: An observational study in a tertiary referral NICU. CBF and DeltaCBV changes were assessed in 41 preterm newborn infants with respiratory distress syndrome treated using mechanical ventilation or the CPAP device. RESULTS: Basal chromophore traces enabled DeltaCBV (mL/100 g) changes to be calculated. CBF was calculated in mL/100 g/min from the saturation rise integral and rate of rise [O(2)Hb-HHb]. Median DeltaCBV was 0.07 (range 0.01-0.13) in SIMV group, 0.07 (0.01-0.19) in HFV group and 0.13 (0.10-1.28) in CPAP group. Median CBF was 14.44 (2.70-32.10), 9.20 (2.94-19.58) and 31.69 (13.59-34.93) respectively. A multiple regression model showed a significant correlation between DeltaCBV or CBF and ventilation approach. CONCLUSION: In the light of our results, we might speculate that, assuming that hemodynamic autoregulation is safe and arterial blood pressure is preserved, ventilation per se influences brain circulation. PMID- 19217224 TI - Clinical and echocardiographic diagnosis of patent ductus arteriosus in premature neonates. AB - The ductus arteriosus frequently fails to close in premature neonates. Considerable difference in opinion exists around what signifies a hemodynamically significant patent ductus arteriosus (PDA) and how reliable clinical signs are in determining the degree of the left-to-right shunting. Although reliance on clinical signs alone could delay the diagnosis of a PDA, there is insufficient evidence to suggest that early treatment improves outcome. Echocardiography is often used as the gold standard for diagnosing a PDA. A combination of echocardiographic measurements may assist in the early diagnosis of a PDA with a hemodynamically significant degree of left-to-right shunting, especially in extremely premature babies, where closure can be significantly delayed. Decision to treat PDA should be based on a combination of clinical signs and echocardiographic parameters. Monitoring B-type natriuretic peptide may be useful in the diagnosing neonates with symptomatic PDA. PMID- 19217225 TI - Effect of the apoptosis rate observed in oocytes and cumulus cells on embryo development in prepubertal goats. AB - Oocyte quality is the main factor that determines blastocyst yield; any factor that could affect it, such as apoptosis, could impair subsequent embryonic development. Our aim was to investigate the incidence of apoptosis in prepubertal goat oocytes and cumulus cells, assessed by Annexin-V staining and TUNEL assay, and their effect on embryo development. Oocyte-cumulus complexes (COCs) from slaughtered females were collected and classified depending on COC morphology as: Healthy (H) and Early Atretic (EA). Each one of these groups was classified depending on oocyte diameter: A: 110-125microm, B: 125-135microm and C: >135microm. The COCs were IVM for 27h, IVF with fresh semen and IVC for 8 days after insemination. Apoptosis analyses were performed before and after maturation. Annexin-positive oocytes decreased with diameter in the EA class (immature oocytes: A: 42.6%; B: 30.3%; C: 21%; IVM-oocytes: A: 17.5%; B: 4.8%; C: 0%), while TUNEL assay showed a decrease of apoptosis in the largest oocytes before and after IVM only in Healthy oocytes (immature oocytes: A: 51.5%; B: 43.3%; C: 12.1%; IVM-oocytes: A: 31.7%; B: 12%; C: 0%). Blastocyst rate increased with increasing oocyte diameter, and it was higher in H than in EA oocytes (Healthy; A: 0%; B: 5.3%; C: 14.4%; Early atretic: A: 0.3%; B: 4.1%; C: 5.1%). Oocyte diameter and COC morphology had no effect on the percentage of apoptosis in blastocyst cells. In conclusion, oocyte developmental competence in prepubertal goats is influenced by oocyte diameter and COC morphology. PMID- 19217226 TI - Colloidal centrifugation with Androcoll-E prolongs stallion sperm motility, viability and chromatin integrity. AB - The objective was to investigate the changes in stallion sperm quality (sperm motility, viability, membrane integrity and chromatin integrity) occurring during cool storage, and to study the effect of sperm selection by single layer colloidal centrifugation on these parameters of sperm quality. Spermatozoa from 3 stallions (10 ejaculates, 3-4 per stallion) were selected by centrifugation through a single layer of colloid (SLC). The resulting sperm preparations and the control samples (extended but unselected semen samples) were stored at 5 degrees C for 48h. Assessments of sperm quality, such as sperm motility, viability (SYBR 14/PI staining), membrane stability (Annexin-V/PI staining) and chromatin integrity, were performed on aliquots of the selected sperm preparations and unselected samples on the day of collection (3h) and after 24 and 48h of storage. In the SLC-selected sperm samples, sperm motility, sperm viability, proportions of spermatozoa with normal morphology and with intact chromatin were significantly better than in unselected samples (motility: 77+/-4% vs. 64+/-8% at 3h; P<0.001; viability: 79.5+/-9% vs. 64.7+/-9%, P<0.001; normal morphology 89+/ 6% vs. 69+/-9%; chromatin integrity DFI 11.3+/-5% vs. 22.1+/-10%). Membrane stability, however, was not different in the SLC-selected and unselected samples (74.6+/-8% vs. 69.3+/-8%). The deterioration seen in sperm quality in the unselected samples was prevented by SLC, so that sperm viability, membrane stability and chromatin integrity were unchanged in the selected samples by 48h compared to 3h (P<0.001), whereas the unselected samples were significantly worse by 48h (P<0.001). Furthermore, it should be possible to send an aliquot of a normal insemination dose (i.e. unselected spermatozoa) overnight to a reference laboratory for analysis of both plasma membrane and chromatin integrity. In conclusion, centrifugation of stallion spermatozoa through a single layer of colloid is a useful technique for selecting the best spermatozoa from an ejaculate and, moreover, sperm quality is maintained during storage. PMID- 19217227 TI - Allelic switching of the imprinted IGF2R gene in cloned bovine fetuses and calves. AB - Cloned animals often suffer from loss of development to term and abnormalities, typically classified under the umbrella term of Large Offspring Syndrome (LOS). Cattle are an interesting species to study because of the relatively greater success rate of nuclear transfer in this species compared with all species cloned to date. The imprinted insulin-like growth factor receptor (IGF2R; mannose-6 phosphate) gene was chosen to investigate aspects of fetal growth and development in cloned cattle in the present study. IGF2R gene expression patterns in identical genetic clones of several age groups were assessed in day 25, day 45, and day 75 fetuses as well as spontaneously aborted fetuses, calves that died shortly after birth and healthy cloned calves using single stranded conformational polymorphism gel electrophoresis. A variable pattern of IGF2R allelic expression in major organs such as the brain, cotyledon, heart, liver, lung, spleen, kidney and intercotyledon was observed using a G/A transition in the 3'UTR of IGF2R. IGF2R gene expression was also assessed by real time RT-PCR and found to be highly variable among the clone groups. Proper IGF2R gene expression is necessary for survival to term, but is most likely not a cause of early fetal lethality or an indicator of postnatal fitness. Contrary to previous reports of the transmission of imprinting patterns from somatic donor cells to cloned animals within organs in the same cloned animal the paternal allele of IGF2R can be imprinted in one tissue while the maternal allele is imprinted in another tissue. This observation has never been reported in any species in which imprinting has been studied. PMID- 19217228 TI - Analysis of organic volatile residues in 9 mm spent cartridges. AB - Determining the time since discharge of spent cartridges found on a crime scene may be very useful in firearm investigations. The potential of small calibre munitions was barely studied in the past and this work did therefore focus on that problematic. The first step was to optimise the detection potential of solid phase microextraction (SPME) followed by gas chromatography coupled to a mass spectrometry detector (GC/MS). This allowed determining the organic volatile composition of empty cartridges immediately after a gunshot. Identification of 32 detected compounds was confirmed by the analysis of reference substances. Preliminary aging studies over 32 h were carried out on selected target compounds to evaluate their potential to determine the time since discharge of empty cartridge cases. PMID- 19217229 TI - Clinical overview of children with mucopolysaccharidosis type III A and effect of Risperidone treatment on children and their mothers psychological status. AB - Mucopolysaccharidosis IIIA (MPS IIIA) is a lysosomal storage disorder characterized by progressive mental deterioration and severe behavioral problems. We conducted an open-label, crossover study of the efficacy and safety of Risperidone on behavioral disorder in children with MPS IIIA. A total of 12 patients (5.5+/-2.2 years) with enzymatic diagnosis of MPS IIIA were randomly assigned to receive Risperidone (0.125-2mg/d) for 6 months. The hyperactivity and disruptive behavior level of children before and after treatment was evaluated regarding the scores from Turgay DSM IV Based Child and Adolescent Behavior Disorders Screening and Rating Scale (T-DSM-IV-S). Clinic Global Impression Scale - Severity (CGIS-S) was used for all cases for determining the psychiatric disorder severity. The anxiety and depression levels of mothers before and after treatment were evaluated using Hamilton Anxiety Scale (HAM-A) and Beck Depression Inventory (BDI). The adverse effects were evaluated by monitoring weight, serum prolactin, glucose and lipid levels. The response to the treatment was measured by decrease in values of CGI-S (from 6+/-1.12 to 2.91+/-0.66, p=0.001). According to T-DSM-IV-S scores the best improvement was observed in hyperactivity scores (16.25+/-8.57/11.58+/-7.26, p=0.001), followed by opposition/defiance (6.66+/ 5.92/5.08+/-4.88, p=0.032), and conduct disorder scores (1.00+/-1.85/0.41+/-.99, p=0.67). No clinically relevant elevations in weight and serum prolactin, glucose or lipid levels have been documented (p>0.05). There was a significant decrease in anxiety and depression scores of mothers (HAM-A: 20.33+/-8.28/17.91+/-6.89, BDI: 23.58+/-7.14/20.5+/-5.93, p<0.001). To our knowledge, research on the pharmacological treatment of MPS IIIA with Risperidone has not been reported. According to our data, Risperidone appeared to be safe and effective in MPS IIIA patients. PMID- 19217231 TI - Long-term follow-up of pyogenic liver abscess by ultrasound. AB - BACKGROUND/AIMS: To study the morphology of residual abscess in successfully treated pyogenic liver abscess by ultrasound. METHODS: 116 consecutive patients of liver abscess diagnosed from April 2004 to March 2007 were included in this study. Patients with amebic liver abscess were excluded. Patients were treated with antibiotics for 6 weeks and when indicated drained percutaneously. Ultrasound scan was repeated after 2 weeks, 1 month, 3 months, 6 months and 12 months of treatment. Thereafter it was repeated every 6 months. RESULTS: After exclusion, 102 patients with >150 lesions were studied. Diagnosis was made on the basis of clinical presentation, ultrasound examination and diagnostic aspiration. It was single in 83, double in 12 and more than 2 in seven patients. Size varied from 4.2cm to 15cm. Organisms were isolated in 29 patients (E. coli 10, Kliebsiella sp. 6, S. aureus 5, Pseudomonas aeruginosa 2, Acenetobacter 1 and multiple growth 5) in blood and eight patients (E. coli 5 and S. aureus 3 and multiple growth 2) in pus. Majority of abscess resolved sonologically after 2-18 weeks of treatment. Healing was delayed in alcoholic and diabetic patients. In eight patients there was residual abscess even after 104 weeks of follow-up. In four patients it healed with calcification. CONCLUSIONS: Majority of pyogenic liver abscess resolve to normal parenchyma within 18 weeks time. However, some lesions may take longer time to heal even after successful treatment. It should be considered in differential diagnosis of space occupying lesion of the liver in ultrasound and need no therapeutic intervention. PMID- 19217230 TI - Is oxidative damage in operation in patients with hereditary spastic paraparesis? AB - Oxidative stress resulting from increased free radical production and/or defects in antioxidant defences may be the cause of various neurodegenerative disorders. In this study, the roles of oxygen free radicals, nitric oxide, superoxide dismutase, vitamin E and vitamin C were investigated in pure and complicated hereditary spastic paraparesis (HSP) patients. The results showed that plasma SOD, vitamin E and nitric oxide levels were significantly low in HSP patients. These findings indicate the influence of oxidative damage in the degenerative process of HSP. PMID- 19217232 TI - Fluorescence in situ hybridization for intracellular localization of nifH mRNA. AB - Few reports on in situ mRNA detection in bacteria have been published, even though a major aim in environmental microbiology is to link function/activity to the identity of the organisms. This study reports a reliable approach for the in situ detection of nifH mRNA using fluorescence hybridization based on a previously described protocol for pmoA. nifH codes for a dinitrogenase reductase, a key enzyme in dinitrogen fixation. nifH mRNA was hybridized with a digoxigenin labelled polynucleotide probe. The hybrid was detected with an anti-DIG-antibody labelled with horseradish peroxidase. Subsequently, the signal was amplified by catalyzed reporter deposition (CARD) with fluorochrome-labelled tyramides. Furthermore, the imaged organisms were identified using standard fluorescence in situ hybridization of rRNA. Thus, the approach enabled us specifically to link in situ the information from the dinitrogen fixation activity of an organism to its identity. Unexpectedly, the signals derived from nifH mRNA hybridization showed a distinct uneven pattern within the cells. This indicated that the method used could even give insights about the localization of the detected mRNA within the cell, which is a potential use of mRNA fluorescence in situ hybridization (FISH) that has not been reported up to now for bacterial cells. PMID- 19217233 TI - Diversity of aerobic and facultative alkalitolerant and halotolerant endospore formers in soil from the Alvord Basin, Oregon. AB - Many proteins produced by Bacillus species isolated from extreme environments have been utilized for industrial purposes, as these extreme environments often promote evolution of unique protein properties. The Borax Lake area is unusual due to its geothermal activity, elevated pH, and high arsenic and salt concentrations in its soils. Soils from this region are likely to harbor alkalitolerant, halotolerant, endospore-forming strains that may be of potential ecological and/or commercial interest. The objectives of this study were to develop new PCR primers that could target Bacillus or closely related 16S rRNA genes, to characterize the diversity of alkalitolerant, halotolerant, endospore forming organisms in the soils surrounding Borax Lake, and to identify novel organisms that may ultimately provide new enzymes for applied use. A three pronged approach was used to identify such bacteria in soil samples. Organisms were isolated using two different techniques. Finally, metagenomic DNA from soil samples was subjected to 16S rRNA gene amplification using the newly designed primers. Assays were performed to characterize the halotolerance and alkalitolerance of isolates. Four different endospore-forming genera and 22 different species were identified by sequencing their 16S rRNA genes. Twenty-five organisms had 96% or less identity to known organisms. Thus, the newly designed Bacillus-related PCR primer sets proved useful for the detection of new species of endospore-forming bacteria in these unique soils. Results indicate that the collection of strains obtained from the Borax Lake region represents a rich source of alkalitolerant, halotolerant, endospore formers. PMID- 19217234 TI - Plasma pharmacokinetics, tissue distribution and excretion study of 6-gingerol in rat by liquid chromatography-electrospray ionization time-of-flight mass spectrometry. AB - A rapid resolution liquid chromatography coupled with electrospray ionization (ESI) time-of-flight mass spectrometry method was developed and validated for quantitative analysis of 6-gingerol in plasma and various tissues. Liquid-liquid extraction was employed as sample preparation technique. Biological samples were separated on an Agilent Zorbax StableBond-C(18) column (4.6 mm x 50 mm, 1.8 microm) and detected by TOF/MS with electrospray ionization (ESI) interface in positive ion mode. Calibration curves (1/x(2) weighted) offered satisfactory linearity (r(2)>0.995) within the test range. The lower limit of quantification in different matrices was in a range of 10-100 ng/mL. Inter- and intra-day precision were in the range of 0.91-11.90% and 0.75-10.23%, respectively. Recoveries in plasma, urine and tissues ranged from 72.5% to 90.4%. Glucuronide of 6-gingerol, the major metabolite of 6-gingerol, was further determined after beta-glucuronidase hydrolyzation. This developed method was successfully applied to pharmacokinetics, tissue distribution and excretion studies of 6-gingerol after oral or intraperitoneal administration in rats. PMID- 19217235 TI - Cervical spondylodiscitis, osteomyelitis, and epidural abscess mimicking a vertebral fracture. AB - BACKGROUND: Spinal vertebral fractures are common in elderly patients, especially after a significant fall. Neck tenderness or neurologic deficits after a fall should increase the suspicion of cervical spinal fracture. CASE REPORT: A 75-year old woman complained of 2 weeks of gradually increasing neck and right shoulder pain after a fall. She had an X-ray study that appeared to show an unstable C6-C7 fracture. A computed tomography scan of the neck revealed no fracture and was consistent with spondylodiscitis and osteomyelitis of C6 and C7, with an adjacent epidural abscess and mild cord compression. CONCLUSION: It is important to consider this diagnosis in patients with a history of infective endocarditis who develop back pain, to minimize morbidity associated with this disorder. PMID- 19217236 TI - Using a laryngeal tube suction-device (LTS-D) reduces the "no flow time" in a single rescuer manikin study. AB - BACKGROUND: In 2005, the European Resuscitation Council and the American Heart Association published new guidelines for Advanced Life Support. One of the points was to reduce the time without chest compressions in the first phase of cardiac arrest. OBJECTIVE: We evaluated in a manikin model whether using the single-use laryngeal tube with suction option (LTS-D) instead of endotracheal intubation (ET) and bag-mask-valve ventilation (BMV) for emergency airway management could reduce the "no-flow time" (NFT). The NFT is defined as the time during resuscitation when no chest compressions take place. METHODS: A randomized, prospective study was undertaken with 150 volunteers who performed management of a standardized simulated cardiac arrest in a manikin. Every participant was randomized to one of three different airway management groups (LTS-D vs. ET vs. BMV). RESULTS: The LTS-D was inserted significantly faster than the ET tube (15 s vs. 44 s, respectively, p < 0.01). During the cardiac arrest simulation, establishing and performing ventilation took an average of 57 s with the LTS-D compared to 116 s with ET and 111 s with the BMV. Using the LTS-D significantly reduced NFT compared to ET and the BMV (125 s vs. 207 s vs. 160 s; p < 0.01). CONCLUSIONS: In our manikin study, NFT was reduced significantly when the LTS-D was used when compared to ET and BMV. The results of our manikin study suggest that for personnel not experienced in tracheal intubation, the LTS-D offers a good alternative to ET and BMV to manage the airway during resuscitation, and to avoid the failure to achieve tracheal intubation with the ET, and the failure to achieve adequate ventilation with the BMV. PMID- 19217237 TI - Use of biomarkers in triage of patients with suspected stroke. AB - BACKGROUND: The absence of a rapidly available and sensitive diagnostic test represents an important limitation in the triage of patients with suspected stroke. OBJECTIVES: The aim of the present study was to investigate the triage accuracy of a novel test that measures blood-borne biomarkers (triage stroke panel, TSP) and to compare its accuracy with that of the Cincinnati Prehospital Stroke Scale (CPSS). METHODS: Consecutive patients with suspected stroke presenting to the Emergency Departments of three Italian hospitals underwent triage by a trained nurse according to the CPSS and had blood drawn for TSP testing. The TSP simultaneously measures four markers (B-type natriuretic peptide, D-dimer, matrix metalloproteinase-9, and S100beta) presenting a single composite result, the Multimarker Index (MMX). Stroke diagnosis was established by an expert committee blinded to MMX and CPSS results. RESULTS: There were 155 patients enrolled, 87 (56%) of whom had a final diagnosis of stroke. The area under the receiver operating characteristic (ROC) curve for CPSS was 0.77 (95% confidence interval [CI] 0.70-0.84) and that of MMX was 0.74 (95% CI 0.66-0.82) (p = 0.285). Thus, both tests, when used alone, failed to recognize approximately 25% of strokes. The area under the ROC curve of the combination of the two tests (0.86, 95% CI 0.79-0.91) was significantly greater than that of either single test (p = 0.01 vs. CPSS and p < 0.001 vs. TSP). CONCLUSIONS: In an emergency care setting, a panel test using multiple biochemical markers showed triage accuracy similar to that of CPSS. Further studies are needed before biomarkers can be introduced in the clinical work-up of patients with suspected stroke. PMID- 19217238 TI - A living will misinterpreted as a DNR order: confusion compromises patient care. AB - BACKGROUND: Advance directives are becoming ever more commonplace in the United States. Correct interpretation of living wills and do-not-resuscitate (DNR) orders is essential if patient safety and autonomy are to be preserved. OBJECTIVES: 1) To recount a case in which a living will was misinterpreted as a DNR order; 2) To make known the ramifications of this misinterpretation; 3) To advocate for improved education of health care professionals regarding the interpretation and implementation of advance directives. CASE REPORT: Mr. S. is an 89-year-old nursing home resident who agreed to the terms of a living will. This living will was subsequently misinterpreted as a DNR order by the patient's physician. This misinterpretation set off a cascade of events that led to the completion of an out-of-hospital DNR order and a compromise of patient care. CONCLUSION: This case study underscores the potential for misunderstanding of an advance directive and the consequent effect on patient care. Likely this is the result of a fundamental lack of understanding about the terminology and definitions inherent in an advance directive document. PMID- 19217239 TI - Pericardial effusion and pneumothorax after breast augmentation. PMID- 19217240 TI - Suicide attempt by intravenous injection of gasoline: a case report. AB - BACKGROUND: There is much experience with intoxication by aspiration of volatile hydrocarbon products, whereas intravenous injection of these distillates is rare. There are only few reports that describe a wide variety of associated pathological changes, predominantly in the pulmonary system. OBJECTIVES: We report the case of an intravenous self-injection of gasoline by a young man in a suicide attempt. CASE REPORT: Immediately after injecting gasoline, the 22-year old man developed bradycardia, hypotension, and increasing dyspnea. Computed tomography scan of the chest showed signs consistent with diffuse alveolar-toxic damage to the lung. These symptoms and radiological findings are similar to those commonly observed after inhalation of this type of substance. This may have been due to diffusion of gasoline into the alveoli, where its presence leads to this characteristic damage. In this patient, gasoline entered the intramuscular tissue, and the patient developed a soft-tissue phlegmon at the forearm. At operation, gas emanation and superficial necrosis were noted. Nevertheless, the patient's outcome was good, with full recovery within 3 weeks. CONCLUSIONS: The major changes in this patient after intravenous injection of gasoline were in the pulmonary system, including hypoxemia and radiological findings that could be related to an exhalation of the volatile substance. In addition, gas in the musculature of the injection area caused a soft-tissue phlegmon. PMID- 19217241 TI - Intrathoracic hiatal hernia. PMID- 19217242 TI - Resuscitation, prolonged cardiac arrest, and an automated chest compression device. AB - BACKGROUND: The European Resuscitation Council's 2005 guidelines for cardiopulmonary resuscitation (CPR) emphasize the delivery of uninterrupted chest compressions of adequate depth during cardiac arrest. OBJECTIVES: To describe how the circumstances of out-of-hospital cardiac arrest can impede the performance of CPR, and how this situation can be overcome. CASE REPORT: The presentation of two cases of prolonged CPR (48 min and 120 min, respectively) with an automated chest compression device, the AutoPulse, under difficult circumstances. Both patients survived without neurological sequelae. CONCLUSION: Prolonged chest compressions may be necessary in some cardiac arrests. These cases suggest that automated chest compression devices may increase the chance of a favorable outcome in these rare situations. PMID- 19217243 TI - The effect of alpha-blockade in emergency department patients with ureterolithiasis. AB - BACKGROUND: Recent studies suggest that alpha-blockade with tamsulosin may be useful in the treatment of ureterolithiasis by increasing the success of, and decreasing the time to, stone passage. OBJECTIVES: We sought to determine stone passage success rates in patients diagnosed with ureterolithiasis and discharged from our Emergency Department (ED) with tamsulosin. METHODS: We conducted a non randomized retrospective cohort study over a 1-year period during which we made monthly follow-up telephone calls to patients discharged from our ED with a diagnosis of ureterolithiasis determined by computed tomography (CT) scan. Stone size and location (proximal, middle, or distal ureter) were determined from radiologist reports and prescriptions were determined from electronic medical records. Patients were asked if they spontaneously passed their stone, how much time was required before stone passage, and if any subsequent hospital visit was required after discharge due to pain or complication resulting from the ureteral stone. RESULTS: During the study, a total of 213 patients were discharged from the ED with ureterolithiasis diagnosed by CT scan; of these, 119 (56%) were successfully contacted and 113 (53%) agreed to participate. Of the 113 patients, 83 (73%) had been prescribed tamsulosin and 30 (27%) had not. Overall age (43 years), gender (67% male), median stone size (3 mm, interquartile range 2-4 mm), and location (71% distal ureter) were similar in the two cohorts. Of tamsulosin recipients, 84% (95% confidence interval [CI] 75-91%) reported passing their stone, whereas 60% (95% CI 42-75%) of non-recipients passed their stone. CONCLUSION: Patients with ureterolithiasis discharged from our ED and given tamsulosin by prescription had moderately high ureteral stone passage success rates in our non-randomized retrospective cohort study. Compared to non recipients of tamsulosin, patients receiving tamsulosin seemed to have greater success in stone passage. PMID- 19217244 TI - Fire jumpers: description of burns and traumatic injuries from a spontaneous mass gathering and celebratory riot. AB - BACKGROUND: On April 3 and 5 of 2005, approximately 52,000 people gathered in Chapel Hill, North Carolina to celebrate the National Collegiate Athletic Association Final Four victories of the University of North Carolina men's basketball team. As crowds rejoiced and intensified into a celebratory riot, many participants lit dozens of bonfires and expressed themselves by jumping through the flames. OBJECTIVE: To describe the interesting injury mechanism of celebratory fire jumping and to describe the injuries associated with two celebratory riots. METHODS: We conducted a cross-sectional study analyzing all Emergency Medical Services (EMS) and hospital reports of injuries associated with each gathering. We used a standardized data collection instrument to record descriptive information on all patients with celebration-associated complaints and noted their treatment and disposition. For analysis, we abstracted data and generated basic descriptive statistics and comparisons of groups. RESULTS: A total of 58 celebrants received medical care, including 27 patients first evaluated by EMS and 49 patients subsequently evaluated in the Emergency Department. Most were young (average age = 23.8 years), male (65%, 32/49), had complaints associated with alcohol (65%, 32/49), and were not admitted to the hospital (92%, 45/49). Of those presenting for hospital evaluation, 30% (15/49) had burns associated with fire-jumping. CONCLUSIONS: Most patients from these gatherings had relatively minor injuries or medical complaints. However, burns associated with fire-jumping represented a substantial proportion of more serious injuries and hospitalizations. These cases are presented to increase awareness of the burn injuries associated with this type of celebratory mass gathering. PMID- 19217245 TI - Comparison of methods for identifying ano-genital injury after consensual intercourse. AB - BACKGROUND: Microtrauma occurs after consensual intercourse. Understanding the injury pattern from consensual intercourse may enlighten our understanding of the ano-genital injury after sexual assault. OBJECTIVE: The purpose of this report is to compare consensual intercourse-related ano-genital injury prevalence by using three different forensic examination techniques: 1) direct visual inspection, 2) colposcopy, and 3) toluidine-blue contrast application. METHODS: Using a descriptive, comparative design, 120 female volunteers, aged 21 years or older, were examined after consensual sexual intercourse using the above techniques. Ano genital injuries were noted using the TEARS classification (Tears, Ecchymoses, Abrasions, Redness, and Swelling). RESULTS: Direct visualization and colposcopy yielded similar ano-genital injury findings. However, more tears and abrasions of the external genitalia were identified with toluidine-blue than with direct visual inspection or colposcopy (p < 0.05). More tears were identified on the anus with toluidine-blue as compared to direct visualization (p < 0.05), but not colposcopy. Fewer ecchymoses were identified on the internal genitalia and fewer areas of redness were identified on both the external and internal genitalia when toluidine-blue was used, as compared to either direct visualization or colposcopy (p < 0.05). CONCLUSIONS: The scientific community needs to continue to build information about ano-genital injury prevalence after consensual sexual intercourse. Understanding the ano-genital injury patterns, including frequency and prevalence, that occur with consensual sexual intercourse will help to identify the difference between injury related to consensual vs. non-consensual sexual intercourse. At this time, toluidine-blue staining may add value to the sexual assault forensic examination. It is the health care provider's role to collect all possible evidence and the courts' role to determine what evidence is admissible and to ensure a rightful conviction. PMID- 19217246 TI - Antidiabetic effect of orally administered conophylline-containing plant extract on streptozotocin-treated and Goto-Kakizaki rats. AB - Conophylline, a vinca alkaloid from Ervatamia microphylla, is known to induce the differentiation of pancreatic precursor cells to insulin-producing cells. In the present research we examined the antidiabetic effects of this alkaloid in vivo by oral administration. Crude conophylline preparations were prepared from the leaves of Tabernaemontana divaricata collected in Okinawa Prefecture, Japan. Conophylline was orally absorbed and showed an increase in its plasma level in both normal and streptozotocin-induced diabetic Sprague-Dawley rats. The plasma conophylline concentration reached its maximum from 1.5 to 3h after a single oral administration and gradually decreased in 24h. The alkaloid decreased the blood glucose level and increased the plasma insulin level in streptozotocin-induced diabetic rats after repetitive administration for 15 days. Fasting blood glucose levels in rats treated orally with conophylline at 0.11 and 0.46 mg/kg/day were 411+/-47 and 381+/-65 mg/dl, respectively; whereas the glucose level of the control rats was 435+/-46 mg/dl. Conophylline also decreased the fasting blood glucose level in Goto-Kakizaki rats in a dose-dependent manner after repetitive administration for 42 days. These results suggest that the extract from conophylline-containing leaves may be useful as a functional food for the treatment of type 2 diabetes mellitus. PMID- 19217247 TI - [Obstructive sleep apnea syndrome, hypertension and artery]. AB - Obstructive sleep apnea syndrome (OSAS), due to upper airway collapse, is frequent but still underestimated. The dose-response relation between OSAS and hypertension (HTN) is now well established. Logically, therefore, blood pressure must be tested in every apneic patient, if necessary by ambulatory blood pressure monitoring. Multiple mechanisms explain this relation, most importantly the increase in sympathetic activity during apnea episodes. OSAS-related hypertension has several characteristics: it is highly prevalent, predominantly diastolic and nocturnal, and frequently affects non-dippers; and the HTN tends to be resistant to treatment. OSAS promotes the formation of arterial lesions (parietal thickening of the carotid artery, increased aortic stiffness, and endothelial dysfunction); the more severe the OSAS, the more severe the lesions. The beneficial effects on blood pressure of continuous positive airway pressure (CPAP), the benchmark treatment for OSAS, are still debated but appear to be significant for untreated or refractory hypertension, for severe OSAS, and when CPAP compliance is good. It also seems promising for the reduction of arterial lesions linked to OSAS. PMID- 19217248 TI - [Tinea faciei, underrecognized because clinically misleading. 14 cases]. AB - INTRODUCTION: The term Tinea Faciei (TF) designates a superficial dermatophyte infection of the glabrous skin of the face, characterized by clinical polymorphism. The aim of the present study is to assess epidemiologic, clinical, mycological, and therapeutic characteristics of TF in a Tunisian dermatology department. PATIENTS AND METHODS: This retrospective study examined the records of cases of TF observed in our dermatology department over the past 5 years (2003 2007). RESULTS: Fourteen patients with TF were seen during the study period. The sex ratio (5 male/9 female) was 0.56. The average age of these 4 children and 10 adults was 22 years. While the clinical form was typical of TF in 10 cases, 4 patients had a clinical form modified by topical steroids (Tinea incognito). Direct microscopic examination showed hyphal elements in 12 cases, and the mycological culture was positive for 11. Five patients had Trichophyton Violaceum, and 4 T. Mentagrophytes. DISCUSSION: TF is frequently misdiagnosed because of its variable appearance. Atypical forms known as tinea incognito are observed after modification of the dermatophyte infections by corticosteroid treatment. Our study included 4 cases of Tinea incognito. T. Mentagrophytes and T. Rubrum are the agents reported most frequently. Unusually, T. Violaceum was found most frequently in our series. PMID- 19217249 TI - [Crohn colitis complicated by a gastrocolic and a colovesical fistula: diagnosis and therapeutic management]. PMID- 19217250 TI - [Ventilatory support at home: indications of oxygen therapy and non invasive ventilation]. AB - The main indication of LTO is COPD with chronic respiratory failure, where it improves life expectancy. By extension, LTO may be proposed to other etiologies of CRF with chronic documented hypoxemia. Daily duration is at least 15H per day. Non invasive ventilation (NIV) represents facial or nasal ventilation. Consensus indications of NIV are restrictive pulmonary diseases (chest wall and /or parenchmal sequelae, neuromuscular diseases, or obesity hypoventilation syndrome) Real role of NIV in log-term management of COPD remains controversial. PMID- 19217251 TI - [Giant hemangioma of the small bowel: a rare cause of intestinal obstruction]. PMID- 19217252 TI - Increasing community integration and inclusion for people with intellectual disabilities. AB - Historically residential facilities for individuals with intellectual disabilities have served the role of segregation and congregation with no real focus on integration into the community. More recently the focus has been to get people out of residential institutions and into community-based living settings. This work examines an approach to changing the systems and culture at a large residential facility to create higher rates of transitions to community-based living settings. A multi-phased systematic implementation approach is discussed in which each successive phase builds upon the previous phase. This approach creates opportunities for community integrated activities and then utilizes these community contexts as functional learning opportunities. Results are evaluated in the areas of community presence, community participation, community integration and community inclusion. Data indicate significant increases in each of these areas based on changing the facility focus, simplifying the intrusive accountability systems, aligning resources and teaching staff how to utilize support plans more efficiently to teach skills in functionally appropriate community integrated activities. PMID- 19217253 TI - Cortical basis of communication: local computation, coordination, attention. AB - Human communication emerges from cortical processing, known to be implemented on a regular repetitive neuronal substratum. The supposed genericity of cortical processing has elicited a series of modeling works in computational neuroscience that underline the information flows driven by the cortical circuitry. In the minimalist framework underlying the current theories for the embodiment of cognition, such a generic cortical processing is exploited for the coordination of poles of representation, as is reported in this paper for the case of visual attention. Interestingly, this case emphasizes how abstract internal referents are built to conform to memory requirements. This paper proposes that these referents are the basis for communication in humans, which is firstly a coordination and an attentional procedure with regard to their congeners. PMID- 19217254 TI - A narrative review of dexterity assessments. AB - NARRATIVE REVIEW: This article is a narrative review of the psychometric properties (reliability and validity) and other characteristics (cost, time to administer, and year of publication) of commercially available manual and finger dexterity assessments used for adults in the United States. Complete research articles related to dexterity assessments were gathered from online database searches and individually critiqued for scientific rigor based on reliability and validity. Articles relating to 14 dexterity assessments were reviewed. All but three tools had established reliability, seven tools had all five forms of validity established, and two had only face and content validity. The results of this review provide information to those interested in fine motor skill acquisition, impairment, or functional recovery after injury. Therapists may use this information to choose the best assessment instrument to evaluate a patient's recovery of function over time. This review adds to the evidence-based, best practice literature related to assessment and outcome measurements of patients with limited dexterity function participating in rehabilitation. PMID- 19217255 TI - Neuroimaging characteristics of patients with focal hand dystonia. AB - NARRATIVE REVIEW: Advances in structural and functional imaging have provided both scientists and clinicians with information about the neural mechanisms underlying focal hand dystonia (FHd), a motor disorder associated with aberrant posturing and patterns of muscle contraction specific to movements of the hand. Consistent with the hypothesis that FHd is the result of reorganization in cortical fields, studies in neuroimaging have confirmed alterations in the topography and response properties of somatosensory and motor areas of the brain. Noninvasive stimulation of these regions also demonstrates that FHd may be due to reductions in inhibition between competing sensory and motor representations. Compromises in neuroanatomical structure, such as white matter density and gray matter volume, have also been identified through neuroimaging methods. These advances in neuroimaging have provided clinicians with an expanded understanding of the changes in the brain that contribute to FHd. These findings should provide a foundation for the development of retraining paradigms focused on reversing overlapping sensory representations and interactions between brain regions in patients with FHd. Continued collaborations between health professionals who treat FHd and research scientists who examine the brain using neuroimaging tools are imperative for answering difficult questions about patients with specific movement disorders. PMID- 19217256 TI - Strengthening the reporting of genetic association studies (STREGA): an extension of the strengthening the reporting of observational studies in epidemiology (STROBE) statement. AB - Making sense of rapidly evolving evidence on genetic associations is crucial to making genuine advances in human genomics and the eventual integration of this information in the practice of medicine and public health. Assessment of the strengths and weaknesses of this evidence, and hence, the ability to synthesize it, has been limited by inadequate reporting of results. The STrengthening the REporting of Genetic Association (STREGA) studies initiative builds on the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) statement and provides additions to 12 of the 22 items on the STROBE checklist. The additions concern population stratification, genotyping errors, modeling haplotype variation, Hardy-Weinberg equilibrium, replication, selection of participants, rationale for choice of genes and variants, treatment effects in studying quantitative traits, statistical methods, relatedness, reporting of descriptive and outcome data, and the volume of data issues that are important to consider in genetic association studies. The STREGA recommendations do not prescribe or dictate how a genetic association study should be designed, but seek to enhance the transparency of its reporting, regardless of choices made during design, conduct, or analysis. PMID- 19217257 TI - Precision of cortical bone reconstruction based on 3D CT scans. AB - The precision and accuracy of human cortical bone reconstruction using 3D CT scans was evaluated using machined bone segments. Both linear and angular errors were measured. Cadaver adult femoral and tibial cortical bone segments were obtained and machined in six orthogonal planes with a precision milling machine. CT scans were then obtained and the bone segments were reconstructed as digital replicas. Dimensional and angular measurements errors were evaluated for the machined bone segments and the results were compared with known dimensions based on milling machine settings to calculate errors due to scanning and model reconstruction. The model dimensional error in the coronal, sagittal and axial directions had a mean of 0.21 mm, with standard a deviation of 0.12 mm and a maximum error of 0.47 mm. The mean percent error was 0.74% and the maximum percent error was 1.9%. The angular error of models in the coronal, sagittal and axial directions was calculated, yielding a mean of 0.47 degrees with a standard deviation of 0.37 degrees and a maximum of 1.33 degrees. The error in the cross sectional axial direction had a mean of 0.54 mm with a maximum error of 0.83 mm, depending on the slice interval. The main error source was of the image processing, which was about 70% of the total error. We found that machining cortical bone segments prior to CT scanning is an effective method for accuracy evaluation of CT-based bone reconstruction. This method can provide a reference for assessing the sensitivity, reliability and accuracy of CT-based applications in the study of movement, finite element modeling, and prosthesis construction. PMID- 19217258 TI - Marginal mass islet transplantation with autologous mesenchymal stem cells promotes long-term islet allograft survival and sustained normoglycemia. AB - Allogeneic islet transplantation is an option to treat diabetes however there are obstacles that are limiting its clinical use. We have examined whether mesenchymal stem cells (MSC) improve islet graft survival and whether such therapy allows for better graft acceptance with reduced requirement for immunosuppression. In vitro-expanded syngeneic bone marrow-derived MSC were co transplanted with islets into omental pouch in a rat model of streptozotocin induced diabetes. Marginal mass syngeneic islet transplantation into the omentum with MSC promoted sustained normoglycemia. Interestingly, allogeneic islets +MSC, but not islets alone, with short-term use of immunosuppression enhanced long-term islet graft survival, insulin expression in the grafts and induced normal serum insulin levels and normoglycemia. T cells from recipients transplanted with allogeneic islets +MSC produced low levels of IFN-gamma and TNF-alpha upon ex vivo activation, and this transplantation protocol promoted the generation of IL 10-secreting CD4(+) T cells. These data encourage further preclinical and eventually, clinical MSC-based islet transplantation to improve the outcome of allogeneic islet transplantation in the treatment of diabetes. PMID- 19217259 TI - Vitamin A deficiency increases hepcidin expression and oxidative stress in rat. AB - OBJECTIVE: The interaction between vitamin A and iron status has been widely reported; however, the exact mechanism involved in this interaction has not been well characterized. The present study investigated the mechanism involved in tissue iron accumulation and changes in the oxidative status in vitamin A deficient rats. METHODS: Rats were treated with a control diet, a vitamin A deficient diet, or a vitamin A/iron-deficient diet for 57 d. The animals were sacrificed; the blood, liver, and spleen were collected for biochemical analysis. Analysis of variance or Mann-Whitney tests were used to compare groups and Pearson's or Spearman's tests to investigate the bivariate correlation. RESULTS: Vitamin A deficiency increased liver hepcidin mRNA and iron spleen concentrations; however, iron deficiency in vitamin A-deficient rats deeply inhibits liver hepcidin mRNA expression and significantly increases divalent metal transporter-1 mRNA levels. Liver ferroportin and hereditary hemochromatosis gene mRNA levels did not change in either treatment. In the vitamin A-deficient groups, liver carbonyl protein increased, whereas catalase and glutathione S transferase activities decreased, suggesting that vitamin A protects the liver against protein oxidation. A significant positive correlation was found between lipid oxidative damage and iron concentration in the liver and spleen (r = 0.611, P = 0.007; r = 0.558, P = 0.025, respectively). CONCLUSION: These results suggest that vitamin A maintains iron homeostasis by the modulation of liver hepcidin expression. The increase of lipid peroxidation in vitamin A deficiency seems to be iron dependent, whereas protein oxidation is not. PMID- 19217260 TI - Postprandial metabolism with 1,3-diacylglycerol oil versus equivalent intakes of long-chain and medium-chain triacylglycerol oils. AB - OBJECTIVE: We assessed the ability of novel lipid structures including medium chain triacylglycerols (MCTs) and 1,3-diacylglycerol (DG) oil to lower postprandial triacylglycerol (TG) elevation and increase hepatic fat oxidation when substituted for dietary TG, which may be useful in the prevention and treatment of obesity and other related metabolic conditions, such as dyslipidemias. METHODS: This double-blind, randomized, crossover trial evaluated the effects of an oral fat load containing DG or MCTs compared with equivalent intakes of long-chain triacylglycerols (LCTs) on the postprandial metabolic responses of insulin-resistant men and women (n = 36). Each subject consumed a single oral fat load on each test day. The fat loads were delivered in milkshakes that contained 30 g of one of the three test oils. RESULTS: The postprandial TG incremental area under the curve after MCT was 73% lower, and that for DG was 22% lower, compared with the response after LCT oil. The incremental area under the curve values for chylomicron TG were reduced versus LCT by 89% and 28%, respectively, in the MCT and DG conditions. Compared with the LCT treatment, beta hydroxybutyrate concentration was increased after MCT oil, but not after DG. CONCLUSION: These results indicate that dietary DG decreased postprandial triglyceridemia compared with LCT, but to a lesser extent than MCT. PMID- 19217261 TI - Lingual cyst with respiratory epithelium: a histopathological and immunohistochemical analysis of two cases. AB - Cysts of the tongue are rare, usually derived from epithelia of the embryonic gastrointestinal and respiratory tracts, and classified according to the predominant epithelium lining. These cysts are usually discovered during infancy, more frequently in males, but they may not appear until well into adulthood. The authors report two lingual cysts lined mainly with respiratory, and focally by squamous, epithelium. Periodic acid-Schiff and mucicarmine staining revealed focal positivity in intracystic mucoid material and goblet cells. Immunohistochemical analysis with vimentin, cytokeratins (AE1/AE3, 34betaE12, CK1, CK5, CK6, CK7, CK8, CK10, CK13, CK14, CK16, CK18, and CK19), E-cadherin, beta-catenin, and epithelial membrane antigen showed a similar profile of normal respiratory epithelium, suggesting well-differentiated states. Owing to their controversial origin, these cysts should be named descriptively, as suggested by Manor et al., as lingual cysts with respiratory epithelium. PMID- 19217262 TI - Oral rehabilitation using pre-shaped Epitec fixation systems after extensive maxillary tumor surgery. AB - Reconstruction of maxillary defects represents a major challenge for reconstructive surgeons. The authors describe two cases of malignant tumor on the maxilla that were successfully repaired using the immediate installation of prostheses supported with an Epitec-system during maxilla reconstruction. The Epitec-plates provided a strong retention source for the maxillodental prosthesis. To reduce the operation time and improve conformity to the remaining bone shape, a craniofacial skull model was fabricated using stereo-lithographic techniques. After trimming the model to simulate segmental resection, Epitec plates were shaped to match the defect. A united Epitec-plate was fixed to the remaining bone immediately and easily after tumor resection. An immediate maxillary prosthesis was placed and was functional at the end of surgery. The Epitec-system provides effective materials for immediate prosthesis of extensive maxillary defects. PMID- 19217263 TI - Human mesenchymal stem cell grafts engineered to release adenosine reduce chronic seizures in a mouse model of CA3-selective epileptogenesis. AB - A novel generation of silk-based brain implants engineered to release adenosine was recently shown to provide robust seizure suppression in kindled rats. As a first step to develop stem cell-coated silk-based 3D-scaffolds for the therapeutic long-term delivery of adenosine we engineered human mesenchymal stem cells (hMSCs) to release adenosine. Here we demonstrate reduction of chronic seizures in a mouse model of CA3-selective epileptogenesis after infrahippocampal transplantation of adenosine-releasing hMSCs. PMID- 19217264 TI - Epigenetics, traffic and firewood. PMID- 19217265 TI - Application of a multifractal analysis to study brain white matter abnormalities of schizophrenia on T2-weighted magnetic resonance imaging. AB - Multifractal analysis provides a precise quantitative description of the structural complexity of white matter (WM) on magnetic resonance imaging (MRI). To test this new technique as an aid to elucidating the pathology of schizophrenia, we examined a multifractal dimension (i.e. Deltaalpha) of WM in schizophrenia patients and their relations to clinical variables. We examined 16 patients with schizophrenia and 16 controls matched for age, sex and handedness. Delta alpha value of WM in the prefrontal and frontoparietal lobes and the corpus callosum (genu and splenium) on T2-weighted MRI was calculated. Delta alpha was not significantly different between groups in either region of interest. However, group-by-side interaction for Deltaalpha was found in the frontoparietal WM; post hoc analysis revealed normal left dominant asymmetry in Deltaalpha for frontoparietal WM in control subjects, which was absent in schizophrenia patients. Furthermore, the patients with schizophrenia had a lower asymmetry coefficient ([R-L]/[R+L]) for Deltaalpha in frontoparietal WM. Relations to clinical symptoms from the Positive and Negative Syndrome Scale, Deltaalpha in corpus callosum, and the asymmetry coefficient in prefrontal WM were correlated with negative and general psychopathology symptom scores. Our results support the left-sided dysfunction hypothesis of schizophrenia and its relation to schizophrenic symptoms. Multifractal analysis reveals abnormal patterns of WM structures in schizophrenia that could be implicated in the disorder's etiology. PMID- 19217266 TI - Diffusion tensor imaging of cocaine-treated rodents. AB - Studies in cocaine-dependent human subjects have shown differences in white matter on diffusion tensor imaging (DTI) compared with non-drug-using controls. It is not known whether the differences in fractional anisotropy (FA) seen on DTI in white matter regions of cocaine-dependent humans result from a pre-existing predilection for drug use or purely from cocaine abuse. To study the effect of cocaine on brain white matter, DTI was performed on 24 rats after continuous infusion of cocaine or saline for 4 weeks, followed by brain histology. Voxel based morphometry analysis showed an 18% FA decrease in the splenium of the corpus callosum (CC) in cocaine-treated animals relative to saline controls. On histology, significant increase in neurofilament expression (125%) and decrease in myelin basic protein (40%) were observed in the same region in cocaine-treated animals. This study supports the hypothesis that chronic cocaine use alters white matter integrity in human CC. Unlike humans, where the FA in the genu differed between cocaine users and non-users, the splenium was affected in rats. These differences between rodent and human findings could be due to several factors that include differences in the brain structure and function between species and/or the dose, timing, and duration of cocaine administration. PMID- 19217267 TI - Resonance light scattering spectral method for the determination of serum albumin with the interaction of neutral red-sodium dodecyl sulfonate. AB - Based on the enhancement of resonance light scattering (RLS) of serum albumin interaction with neutral red (NR) and sodium dodecyl sulfonate (SDS), a novel sensitive assay of serum albumins has been developed. Experimental conditions such as mixing sequence of reagents, pH, NR and SDS concentrations have been optimized. Linear relationships between the enhanced RLS intensity and the protein concentration were observed for bovine serum albumin (BSA) within the range of 0.01-5.0 microg mL(-1) and human serum albumin (HAS) of 0.01-7.0 microg mL(-1). The detection limits (S/N=3) are 6.0 ng mL(-1) for BSA and 5.0 ng mL(-1) for HAS, respectively. The method was successfully applied to the determination of HSA in human blood plasma samples with recovery from 97.3 to 104.3%. PMID- 19217268 TI - [Diet for mother and infant and allergy prevention]. PMID- 19217269 TI - MHC molecules and microbial antigen processing in phagosomes. AB - Macrophages and dendritic cells are phagocytic antigen presenting cells that internalize bacteria and other particulate antigens into phagosomes. The phagosome must then balance microbicidal and proteolytic degradation functions with the generation of antigenic peptides for presentation by class I and class II MHC molecules to CD8 and CD4 T cells, respectively. Understanding the host and bacterial factors that affect phagosomal antigen processing may help facilitate new strategies to eliminate pathogens. PMID- 19217270 TI - Altered ventilatory responses to exercise testing in young adult men with obstructive sleep apnea. AB - BACKGROUND: Obstructive sleep apnea (OSA) is a disorder characterized by repetitive obstructions of the upper airway. Individuals with OSA experience intermittent hypoxia, hypercapnia, and arousals during sleep, resulting in increased sympathetic activation. Chemoreflex activation, arising from the resultant oscillatory disturbances in blood gases from OSA, exerts control over ventilation, and may induce increases in sympathetic vasoconstriction, contributing to increased long-term risks for hypertension (HTN) and cardiovascular disease (CVD). METHODS: To evaluate whether OSA elicits exaggerated ventilatory responses to exercise in young men, 14 overweight men with OSA and 16 overweight men without OSA performed maximal ramping cycle ergometer exercise tests. Oxygen consumption (VO(2)), ventilation, (V(E)), ventilatory equivalents for oxygen (V(E)/VO(2)) and carbon dioxide (V(E)/VCO(2)), and V(E)/VCO(2) slope were measured. RESULTS: The VO(2) response to exercise did not differ between groups. The V(E), V(E)/VCO(2), V(E)/VO(2) were higher (p< 0.05, 0.002, and p<0.02, respectively) in the OSA group across all workloads. The V(E)/VCO(2) slope was greater in the OSA group (p<0.05). The V(E)/VCO(2) slope and AHI were significantly correlated (r=0.56, p<0.03). Thus, young, overweight men with OSA exhibit increased ventilatory responses to exercise when compared to overweight controls. This may reflect alterations in chemoreflex sensitivity, and contribute to increased sympathetic drive and HTN risk. PMID- 19217271 TI - Mycobacterium tuberculosis and polymorphonuclear pleural effusion: incidence and clinical pointers. AB - BACKGROUND: Delayed diagnosis and treatment of a polymorphonuclear cell (PMN) predominant pleural effusion due to Mycobacterium tuberculosis (MTB) are associated with poor outcome and the risk of tuberculosis transmission. We investigated the clinical differences of PMN-predominant pleural effusion due to MTB or other microorganisms. METHODS: From January 2000 to April 2007, a total of 354 patients with tuberculous pleurisy were identified. Among them, 39 (11.0%) adults had PMN-predominant pleural effusion (MTB group). Their clinical characteristics were compared with the 117 age-/gender-matched controls (1:3) selected from 715 patients with PMN-predominant pleural effusion due to other microorganisms. RESULTS: Among patients with PMN-predominant septic pleural effusion, 5.2% were due to MTB. The in-hospital mortality rate in the MTB group was 36%, similar to that of the control group. Sputum samples were culture positive for MTB in 41%. Weight loss (p=0.006), initial leukocyte count G (Pro12Ala) and +82466C>T (His449His), in Korean subjects (839 subjects with asthma and 449 normal controls). RESULTS: Association analysis using logistic regression analysis showed that +82466C>T and haplotypes 1(CC) and 2(CT) were associated with the development of asthma (p=0.01-0.04). The frequency of PPARG ht2 was significantly lower in the patients with asthma compared to the normal controls in codominant and dominant models (p=0.01, p(corr)=0.03 and p=0.02, p(corr)=0.03, respectively). Conversely, the frequency of PPARG-ht1 was significantly higher in the patients with asthma compared to the normal controls in the codominant model [p=0.04, OR: 1.27 (1.01-1.6)]. In addition, the rare allele frequency of +82466C>T was significantly lower in patients with asthma in comparison to normal controls in the codominant model (OR: 0.78, p=0.04). Thus, polymorphism of the PPARG gene may be linked to an increased risk of asthma development. PMID- 19217274 TI - The interplay of structural information and functional studies in kinase drug design: insights from BCR-Abl. AB - As an inhibitor of the tyrosine kinase activity of the BCR-Abl oncoprotein, imatinib sets a new paradigm for the treatment of cancer with molecularly targeted therapies. Subsequent structural studies have provided in depth knowledge of how this antileukaemia drug interacts with the catalytic site of the enzyme and allowed the rationalisation of mechanisms of drug-resistance which can lead to patient relapse. This understanding has facilitated the design of new inhibitors of BCR-Abl, as well as the discovery of inhibitors of many other kinases. As structural information accumulates for more of the 518 kinases encoded within the human genome, the design of many more highly selective, well tolerated kinase inhibitors should be possible. PMID- 19217275 TI - Study of the solubility and stability of polystyrene wastes in a dissolution recycling process. AB - Dissolution with suitable solvents is one of the cheapest and more efficient processes for polystyrene waste management. In this work the solubility of polystyrene foams in several solvents benzene, toluene, xylene, tetrahydrofuran, chloroform, 1,3-butanediol, 2-butanol, linalool, geraniol, d-limonene, p-cymene, terpinene, phellandrene, terpineol, menthol, eucalyptol, cinnamaldheyde, nitrobenzene, N,N-dimethylformamide and water has been determined. Experimental results have shown that to develop a "green process" the constituents of essential oils, d-limonene, p-cymene, terpinene, phellandrene, are the most appropriate solvents. The action of these solvent does not produce any degradation of polymer chains. The solubility of the polymer in the mentioned solvents at different temperatures has been investigated. The solvent can be easily recycled by distillation. PMID- 19217273 TI - Cell fate regulation by coupling mechanical cycles to biochemical signaling pathways. AB - Many aspects of cellular motility and mechanics are cyclic in nature such as the extension and retraction of lamellipodia or filopodia. Inherent to the cycles of extension and retraction that test the environment is the production of mechano chemical signals that can alter long-term cell behavior, transcription patterns, and cell fate. We are just starting to define such cycles in several aspects of cell motility, including periodic contractions, integrin cycles of binding and release as well as the normal oscillations in motile activity. Cycles of local cell contraction and release are directly coupled to cycles of stressing and releasing extracellular contacts (matrix or cells) as well as cytoplasmic mechanotransducers. Stretching can alter external physical properties or sites exposed by matrix molecules as well as internal networks; thus, cell contractions can cause a secondary wave of mechano-regulated outside-in and internal cell signal changes. In some cases, the integration of both external and internal signals in space and time can stimulate a change in cell state from quiescence to growth or differentiation. In this review we will develop the basic concept of the mechano-chemical cycles and the ways in which they can be described and understood. PMID- 19217276 TI - SPR sensing of small molecules with modified RNA aptamers: detection of neomycin B. AB - We have studied how the modification of the RNA aptamer evolved against neomycin B at 2' position of ribose with a methyl group influences the affinity of the interaction. Using surface plasmon resonance (SPR) and faradaic impedance spectroscopy (FIS) an affinity constant in the muM range was calculated. The results showed that the modification of the aptamer does not significantly alter the affinity of the aptamer for the antibiotic. This finding opens up the possibility of designing modified RNA aptamers resistant to endonucleases without variation of the analytical features. In addition to this, we propose a competitive assay for the detection of neomycin B using SPR as a transduction technique. A range of quantification between 10 nM and 100 microM was obtained, which shows the feasibility of detecting small molecules using aptamers with high sensitivity. PMID- 19217277 TI - ORC proteins: marking the start. AB - The DNA replication apparatus of archaea is more closely related to that of eukaryotes than eubacteria. Furthermore, recent work has shown that archaea, like eukaryotes, have multiple replication origins. Biochemical data are starting to reveal how archaeal origin binding proteins recognise and remodel origin DNA sequences. Crystal structures of archaeal replication origin binding proteins complexed with their DNA targets revealed details of how they interact with origins and showed that they introduce significant deformations of the DNA. Although these recent advances provide insight about the initial interactions of proteins at archaeal replication origins, the molecular mechanisms of origin assembly and firing still remain elusive. PMID- 19217278 TI - Neutropaenia as a prognostic factor in metastatic colorectal cancer patients undergoing chemotherapy with first-line FOLFOX. AB - We retrospectively analysed 153 patients with metastatic colorectal cancer who received FOLFOX with or without bevacizumab as first-line chemotherapy. Several background characteristics and chemotherapy features (grade of neutropaenia, use of bevacizumab or irinotecan, re-introduction of FOLFOX, and tumour progression) as time-varying covariates were analysed as potential prognostic factors. Of the 153 patients, mild neutropaenia (grade 1-2) occurred in 60 patients (39%) and severe neutropaenia (grade 3-4) occurred in 46 patients (30%). The other 47 patients (31%) did not experience neutropaenia. According to a multivariate Cox model with time-varying covariates, hazard ratios (HRs) of death were 0.55 (95% confidence interval (CI), 0.31-0.98; P=0.044) for patients with mild neutropaenia and 0.35 (95% CI, 0.18-0.66; P=0.002) for those with severe neutropaenia. Both mild and severe neutropaenia during chemotherapy are associated with improved survival in patients with MCRC. Prospective trials are required to assess whether dosing adjustments based on neutropaenia may improve chemotherapy efficacy. PMID- 19217279 TI - Is the recent fall in incidence of post-menopausal breast cancer in UK related to changes in use of hormone replacement therapy? AB - There has been a substantial decline in the use of female sex hormone replacement therapy (HRT) in the United Kingdom, particularly by post-menopausal women, since around 2000-2001. Given what is known of the risk of breast cancer in women receiving HRT, the decline in use should have resulted in a decrease in risk, and incidence rates about 14% lower than expected were predicted for the age group 50 59 in 2005. There has been a recent slowing and reversal of the increasing trends in incidence of breast cancer in the age group 45-64. This is most marked at ages 50-59, where rates since 1999 have been decreasing at 0.8% a year, following a long period of sustained increase. It seems probable that these two events are causally related. PMID- 19217281 TI - Comparative QSAR modelling of 2-phenylindole-3-carbaldehyde derivatives as potential antimitotic agents. AB - QSAR modelling was done on some 2-phenylindole-3-carbaldehyde derivatives to find out structural requirements for more active antimitotic agents. Four statistical methods were used to develop models. The results show the importance of ETSA indices, RTSA indices, IC1, SIC4, Jhetv and MSD on the activity. Electrostatic potential charges of atoms, increased surface area, and presence of bulky group along Y-axis and chlorine substitution were also found to be important. PMID- 19217280 TI - 14-O-Heterocyclic-substituted naltrexone derivatives as non-peptide mu opioid receptor selective antagonists: design, synthesis, and biological studies. AB - Mu opioid receptor antagonists have clinical utility and are important research tools. To develop non-peptide and highly selective mu opioid receptor antagonist, a series of 14-O-heterocyclic-substituted naltrexone derivatives were designed, synthesized, and evaluated. These compounds showed subnanomolar-to-nanomolar binding affinity for the mu opioid receptor. Among them, compound 1 exhibited the highest selectivity for the mu opioid receptor over the delta and kappa receptors. These results implicated an alternative 'address' domain in the extracellular loops of the mu opioid receptor. PMID- 19217282 TI - Antiproliferative effects of peracetylated naphthoxylosides. AB - The antiproliferative activity, and the capability of priming of glycosaminoglycan chains, of two series of peracetylated mono- and bis xylosylated dihydroxynaphthalenes have been investigated for normal HFL-1 cells, as well as transformed T24 cells, and compared to the unprotected analogs. Our data show increased antiproliferative activity upon peracetylation, but a loss of selectivity towards T24 cells. PMID- 19217284 TI - 1,3,4-Oxadiazole substituted naphthyridines as HIV-1 integrase inhibitors. Part 2: SAR of the C5 position. AB - The use of a 1,3,4-oxadiazole in combination with an 8-hydroxy-1,6-naphthyridine ring system has been shown to deliver potent enzyme and antiviral activity through inhibition of viral DNA integration. This report presents a detailed structure-activity investigation of the C5 position resulting in low nM potency for several analogs with an excellent therapeutic index. PMID- 19217283 TI - First small molecular inhibitors of T. brucei dolicholphosphate mannose synthase (DPMS), a validated drug target in African sleeping sickness. AB - Drug-like molecules with activity against Trypanosoma brucei are urgently required as potential therapeutics for the treatment of African sleeping sickness. Starting from known inhibitors of other glycosyltransferases, we have developed the first small molecular inhibitors of dolicholphosphate mannose synthase (DPMS), a mannosyltransferase critically involved in glycoconjugate biosynthesis in T. brucei. We show that these DPMS inhibitors prevent the biosynthesis of glycosylphosphatidylinositol (GPI) anchors, and possess trypanocidal activity against live trypanosomes. PMID- 19217285 TI - Synthesis and SAR study of novel pseudo-steroids as potent and selective progesterone receptor antagonists. AB - Synthesis of novel 7-pseudo-steroids 1c has been achieved from trenbolone 3 via an efficient 14 step sequence with overall yields of 10-15%. Various substitutions were incorporated at both the aromatic side chain as well as the D ring. The orientation of aromatic side chain at C10 plays a crucial role for progesterone receptor (PR) activity. Compound 2a (T47D=1nM) with -NMe(2) para to the aromatic group along with spirofurane groups in the D ring was the optimal substitution. All compounds were also evaluated for glucocorticoid receptor (GR) antagonist activities in vivo in a rat and found efficacious in uterine complement C3 assay via the oral route of administrations. PMID- 19217286 TI - N-Benzyl-indolo carboxylic acids: Design and synthesis of potent and selective adipocyte fatty-acid binding protein (A-FABP) inhibitors. AB - Small molecule inhibitors of adipocyte fatty-acid binding protein (A-FABP) have gained renewed interest following the recent publication of pharmacologically beneficial effects of such inhibitors. Despite the potential utility of selective A-FABP inhibitors within the fields of metabolic disease, inflammation and atherosclerosis, there are few examples of useful A-FABP inhibitors in the public domain. Herein, we describe the optimization of N-benzyl-tetrahydrocarbazole derivatives through the use of co-crystal structure guided medicinal chemistry efforts. This led to the identification of a potent and selective class of A-FABP inhibitors as illustrated by N-benzyl-hexahydrocyclohepta[b]indole 30. PMID- 19217287 TI - Development of multitargeted inhibitors of both the insulin-like growth factor receptor (IGF-IR) and members of the epidermal growth factor family of receptor tyrosine kinases. AB - Emerging clinical and pre-clinical data indicate that both insulin-like growth factor receptor (IGF-IR) and members of the epidermal growth factor (EGF) family of receptor tyrosine kinases (RTKs) exhibit significant cross-talk in human cancers. Therefore, a small molecule that successfully inhibits the signaling of both classes of oncogenic kinases might provide an attractive agent for chemotherapeutic use. Herein, we disclose the structure activity relationships that led to the synthesis and biological characterization of 14, a novel small molecule inhibitor of both IGF-IR and members of the epidermal growth factor family of RTKs. PMID- 19217288 TI - Synthesis and structure-activity relationships of novel benzofuran farnesyltransferase inhibitors. AB - A series of benzofuran-based farnesyltransferase inhibitors have been designed and synthesized as antitumor agents. Among them, 11f showed the most potent enzyme inhibitory activity (IC(50)=1.1nM) and antitumor activity in human cancer xenografts in mice. PMID- 19217289 TI - Atropisomeric small molecule Bcl-2 ligands: determination of bioactive conformation. AB - The separation of atropisomeric conformers of 1,2,3,4-tetrahydroisoquinoline amide Bcl-2 ligands allowed the identification of the bioactive conformer which was subsequently confirmed by X-ray crystallography. PMID- 19217290 TI - Mammalian animal models for Duchenne muscular dystrophy. AB - Duchenne muscular dystrophy (DMD) is a fatal neuromuscular disease that affects boys and leads to early death. In the quest for new treatments that improve the quality of life and in the search for a possible definitive cure, the use of animal models plays undoubtedly an important role. Therefore, a number of different mammalian models for DMD have been described. Much knowledge on the molecular mechanisms underlying the disease has arisen from studies in these animals. However, the use of different models does not often allow a direct comparison of results obtained in preclinical trials and therefore hinders a straightforward translational research. In the frame of "TREAT-NMD", a European Network of Excellence addressing the fragmentation in the assessment and treatment of neuromuscular diseases, we compare here the currently used mammalian animal models for DMD with the aim of selecting and recommending the most appropriate ones for preclinical efficacy testing of new therapeutic strategies. PMID- 19217291 TI - The phosphatidylserine receptor TIM-4 does not mediate direct signaling. AB - Engulfment of apoptotic cells is an active process coordinated by receptors on phagocytes and ligands on apoptotic cells [1]. Phosphatidylserine (PtdSer) is a key ligand on apoptotic cells, and recently three PtdSer recognition receptors have been identified, namely, TIM-4, BAI1, and Stabilin-2 [1-6]. Whereas BAI1 is dependent on the ELMO1/Dock180/Rac signaling module, and Stablilin-2 appears to use the intracellular adaptor GULP [2, 3, 7], little is known about how TIM-4 transduces signals downstream of PtdSer recognition [8]. To test the role of known engulfment signaling pathways in TIM-4-mediated engulfment, we used a combination of dominant-negative mutants, knockdown of specific signaling proteins, and knockout cell lines. TIM-4 appears to be largely independent of the two known engulfment signaling pathways [7, 9-17], yet the TIM-4-mediated uptake is inhibited by cytoskeleton disrupting drugs. Remarkably, a version of TIM-4 lacking its cytoplasmic tail promoted corpse uptake via PtdSer recognition. Moreover, replacement of the transmembrane region of TIM-4 with a glycophosphatidylinositol anchor still promoted engulfment comparable to wild type TIM-4. Thus, the transmembrane region and cytoplasmic tail of TIM-4 are dispensable for apoptotic cell engulfment, and we propose that TIM-4 is a PtdSer tethering receptor without any direct signaling of its own. PMID- 19217292 TI - A role for Id2 in regulating photic entrainment of the mammalian circadian system. AB - Inhibitor of DNA binding genes (Id1-Id4) encode helix-loop-helix (HLH) transcriptional repressors associated with development and tumorigenesis [1, 2], but little is known concerning the function(s) of these genes in normal adult animals. Id2 was identified in DNA microarray screens for rhythmically expressed genes [3-5], and further analysis revealed a circadian pattern of expression of all four Id genes in multiple tissues including the suprachiasmatic nucleus. To explore an in vivo function, we generated and characterized deletion mutations of Id2 and of Id4. Id2(-/-) mice exhibit abnormally rapid entrainment and an increase in the magnitude of the phase shift of the pacemaker. A significant proportion of mice also exhibit disrupted rhythms when maintained under constant darkness. Conversely, Id4(-/-) mice did not exhibit a noticeable circadian phenotype. In vitro studies using an mPer1 and an AVP promoter reporter revealed the potential for ID1, ID2, and ID3 proteins to interact with the canonical basic HLH clock proteins BMAL1 and CLOCK. These data suggest that the Id genes may be important for entrainment and operation of the mammalian circadian system, potentially acting through BMAL1 and CLOCK targets. PMID- 19217293 TI - A dynamin is required for the biogenesis of secretory organelles in Toxoplasma gondii. AB - BACKGROUND: Apicomplexans contain only a core set of factors involved in vesicular traffic. Yet these obligate intracellular parasites evolved a set of unique secretory organelles (micronemes, rhoptries, and dense granules) that are required for invasion and modulation of the host cell. Apicomplexa replicate by budding from or within a single mother cell, and secretory organelles are synthesized de novo at the final stage of division. To date, the molecular basis for their biogenesis is unknown. RESULTS: We demonstrate that the apicomplexan dynamin-related protein B (DrpB) belongs to an alveolate specific family of dynamins that is expanded in ciliates. DrpB accumulates in a cytoplasmic region close to the Golgi that breaks up during replication and reforms after assembly of the daughter cells. Conditional ablation of DrpB function results in mature daughter parasites that are devoid of micronemes and rhoptries. In the absence of these organelles, invasion-related secretory proteins are mistargeted to the constitutive secretory pathway. Mutant parasites are able to replicate but are unable to escape from or invade into host cells. CONCLUSIONS: DrpB is the essential mechanoenzyme for the biogenesis of secretory organelles in Apicomplexa. We suggest that DrpB is required during replication to generate vesicles for the regulated secretory pathway that form the unique secretory organelles. Our study supports a role of an alveolate-specific dynamin that was required for the evolution of novel, secretory organelles. In the case of Apicomplexa, these organelles further evolved to enable a parasitic lifestyle. PMID- 19217294 TI - A novel dynamin-related protein has been recruited for apicoplast fission in Toxoplasma gondii. AB - BACKGROUND: Apicomplexan parasites cause numerous important human diseases, including malaria and toxoplasmosis. Apicomplexa belong to the Alveolata, a group that also includes ciliates and dinoflagellates. Apicomplexa retain a plastid organelle (the apicoplast) that was derived from an endosymbiotic relationship between the alveolate ancestor and a red alga. Apicoplasts are essential for parasite growth and must correctly divide and segregate into daughter cells upon cytokinesis. Apicoplast division depends on association with the mitotic spindle, although little is known about the molecular machinery involved in this process. Apicoplasts lack the conserved machinery that divides chloroplasts in plants and red algae, suggesting that these mechanisms are unique. RESULTS: Here, we demonstrate that a dynamin-related protein in Toxoplasma gondii (TgDrpA) localizes to punctate regions on the apicoplast surface. We generate a conditional dominant-negative TgDrpA cell line to disrupt TgDrpA functions and demonstrate that TgDrpA is essential for parasite growth and apicoplast biogenesis. Fluorescence recovery after photobleaching and time-lapse imaging studies provide evidence for a direct role for TgDrpA in apicoplast fission. CONCLUSIONS: Our data suggest that DrpA was likely recruited from the alveolate ancestor to function in fission of the symbiont and ultimately replaced the conserved division machinery of that symbiont. PMID- 19217295 TI - Motion-induced blindness and motion streak suppression. AB - In motion-induced blindness (MIB), persistent static targets intermittently disappear when presented near moving elements [1, 2]. There is currently no consensus regarding the cause or causes of MIB [3-7]. Here, we link the phenomenon to a mechanism that is integral for normal human vision, motion streak suppression [8]. The human visual system integrates information over time [9], resulting in streaks of activity across visual brain regions when objects move [10, 11]. These "motion streaks" are usually suppressed from awareness. Our results suggest that this process shapes MIB. We show that MIB is enhanced at the trailing edges of movement and that both MIB and motion streak suppression are impaired at equiluminance. These findings suggest that an apparent failure of human vision, MIB, is at least partially driven by a functional adaptation that facilitates clear perceptions of moving form. PMID- 19217296 TI - Motor task variation induces structural learning. AB - When we have learned a motor skill, such as cycling or ice-skating, we can rapidly generalize to novel tasks, such as motorcycling or rollerblading [1-8]. Such facilitation of learning could arise through two distinct mechanisms by which the motor system might adjust its control parameters. First, fast learning could simply be a consequence of the proximity of the original and final settings of the control parameters. Second, by structural learning [9-14], the motor system could constrain the parameter adjustments to conform to the control parameters' covariance structure. Thus, facilitation of learning would rely on the novel task parameters' lying on the structure of a lower-dimensional subspace that can be explored more efficiently. To test between these two hypotheses, we exposed subjects to randomly varying visuomotor tasks of fixed structure. Although such randomly varying tasks are thought to prevent learning, we show that when subsequently presented with novel tasks, subjects exhibit three key features of structural learning: facilitated learning of tasks with the same structure, strong reduction in interference normally observed when switching between tasks that require opposite control strategies, and preferential exploration along the learned structure. These results suggest that skill generalization relies on task variation and structural learning. PMID- 19217297 TI - The motor somatotopy of speech perception. AB - Listening to speech recruits a network of fronto-temporo-parietal cortical areas. Classical models consider anterior (motor) sites to be involved in speech production whereas posterior sites are considered to be involved in comprehension. This functional segregation is challenged by action-perception theories suggesting that brain circuits for speech articulation and speech perception are functionally dependent. Although recent data show that speech listening elicits motor activities analogous to production, it's still debated whether motor circuits play a causal contribution to the perception of speech. Here we administered transcranial magnetic stimulation (TMS) to motor cortex controlling lips and tongue during the discrimination of lip- and tongue articulated phonemes. We found a neurofunctional double dissociation in speech sound discrimination, supporting the idea that motor structures provide a specific functional contribution to the perception of speech sounds. Moreover, our findings show a fine-grained motor somatotopy for speech comprehension. We discuss our results in light of a modified "motor theory of speech perception" according to which speech comprehension is grounded in motor circuits not exclusively involved in speech production. PMID- 19217298 TI - Type III secretion systems in symbiotic adaptation of pathogenic and non pathogenic bacteria. AB - The emergence of multi-drug resistance and bacteria with increased virulence is a familiar refrain to the contemporary microbiologist. Although intense research over the past decade has ascribed much molecular detail to these processes, more esoteric questions remain: for example, why are some bacteria evolving increased virulence towards humans, what are the genes underpinning this virulence potential and what are the selective pressures that favor these traits? A holistic approach that considers the organismal biology of bacteria with their diverse hosts seems appropriate to begin to tackle such issues. As it happens, the type III secretion system is turning out to be a central player in the adaptation of both parasites and mutualists to diverse hosts. With this in mind, human interventions in agriculture, animal husbandry and even drug discovery that could influence the selection of bacteria with improved type III secretion system function should be critically appraised. PMID- 19217299 TI - The role of fibroblast growth factor-2 in healing the dura mater after inducing cerebrospinal fluid leakage in rats. AB - We conducted a study to determine the effectiveness of topically applied recombinant mouse fibroblast growth factor-2 (FGF-2) in healing the dura mater in a rat with dura mater injury and cerebrospinal fluid leakage. Laminectomies were performed in 32 rats at the level of the L2-L4 vertebrae, and a dura mater defect was created. Sixteen rats were treated postoperatively with locally applied recombinant mouse FGF-2, and 16 animals received normal saline. FGF-2 effects on dura mater healing, cerebrospinal fluid leakage, and wound healing were assessed at 3 and 6 weeks postoperatively. The extent of dura mater healing was evaluated by histological analysis. We found that dura mater healing was significantly increased (p<0.05) in rats treated with FGF-2 compared with rats in the control group. In this experimental model, locally applied FGF-2 effectively increased dura mater healing and induced no side effects. PMID- 19217300 TI - Beta-catenin gets jaded and von Hippel-Lindau is to blame. AB - Numerous studies have pointed to interactions between the tumor suppressor von Hippel-Lindau (VHL) and the oncogenic Wnt-beta-catenin signaling cascade; however, the mechanism of this crosstalk has remained elusive. Among other roles, VHL can promote the stabilization of Jade-1. Now, recent findings provide compelling evidence that Jade-1 ubiquitylates beta-catenin, leading to its degradation. Thus, the loss of VHL, as seen in clear cell renal cell carcinoma, could lead to tumor formation through beta-catenin de-repression. PMID- 19217301 TI - Synthesis and cytotoxic activity of new azepino[3',4':4,5]pyrrolo[2,1 a]isoquinolin-12-ones. AB - A series of azepino[3',4':4,5]pyrrolo[2,1-a]isoquinolin-12-ones (3a-f), that were conformationally restricted analogs of lead compound 2, were designed as potential cytotoxic compounds and synthesized using a radical oxidative aromatic substitution reaction as the key step. Compounds 3a-f were tested on five tumor cell lines to determine the conformational requirements for biological activity of compound 2. The results show that conformational restrictions on compound 2, generating the derivatives 3a-f, do not appreciably reduce the cytotoxic activity of 2, although compound 3d (R=Br) showed good activity against U-251 cells. Preliminary structure-activity relationship studies with these compounds revealed the importance of halogens bonded to the isoquinoline moiety. Additionally, derivatives 3f (R=NO(2)) and 3b (R=F) were cytotoxic to PC-3 and K-562 cells. However, none of the azepino[3',4':4,5]pyrrolo[2,1-a]isoquinolinones inhibited the enzymatic activity of CDK1/cyclin B, CDK5/p25, or GSK-3. PMID- 19217302 TI - Synthesis of highly deuterium-labeled (R)-K-13675, PPAR alpha agonist, for use as an internal standard for low-level quantification of drugs in plasma. AB - Two highly deuterium-labeled compounds, (R)-K-13675-d(11) and (R)-K-13675-d(7), were prepared for use as internal standards for low-level quantification of plasma drugs by LC/MS/MS. We successfully demonstrated their utility in pharmacokinetic studies for sensitive and precise drug quantification. PMID- 19217303 TI - Effects of the introduction of inversion of polarity sites in the quadruplex forming oligonucleotide TGGGT. AB - Insight into the influence of inversion of polarity sites on the structural features of quadruplex structures is presented. The NMR and CD studies concern modified oligodeoxynucleotides (ODNs) based on the quadruplex forming sequence TGGGT. The presence of inversion of polarity sites not only does not compromise the formation of quadruplexes, but in some cases it increases the thermal stability of modified complexes compared with that of the unmodified one. PMID- 19217304 TI - 5-Arylidene-2-phenylimino-4-thiazolidinones as PTP1B and LMW-PTP inhibitors. AB - As part of a project aimed at identifying effective low molecular weight nonphosphorus monoanionic inhibitors of PTPs, we have synthesized 4-[(5-arylidene 4-oxo-2-phenyliminothiazolidin-3-yl)methyl]benzoic acids (4) and evaluated their inhibitory activity against human PTP1B and LMW-PTP enzymes. The introduction of a 2-phenylimino moiety onto the 4-thiazolidinone ring was designed to enhance the inhibitor/enzyme affinity by means of further favourable interactions with residues of the active site and the surrounding loops. Some of the compounds (4a d, f) showed interesting inhibition levels in the low micromolar range. The 5 arylidene moiety of acids 4 proved to markedly influence the potency of these inhibitors. Molecular modeling experiments inside the binding sites of both enzymes were performed. PMID- 19217305 TI - Synthesis of (-)-lobeline via enzymatic desymmetrization of lobelanidine. AB - The bioactive alkaloid (-)-lobeline was synthesized via the stereoselective acylation (desymmetrization) of meso-lobelanidine by vinyl acetate in the presence of Candida antarctica lipase B. PMID- 19217306 TI - Mitochondrial proteome during salt stress-induced programmed cell death in rice. AB - It has been shown that mitochondria play a pivotal role in plant programmed cell death (PCD). Previous study established a salt stress-induced PCD model in rice (Oryza sativa L. cv. WYJ 8th) root tip cells, demonstrated by DNA laddering, cytochrome c release, and TUNEL positive reaction. In this study, the role of mitochondria during the early phase of PCD (2h-PCD) was analyzed in rice roots. After 2h-PCD induction, the integrity of mitochondria decreased slightly, consistent with a small release of cytochrome c. 2h-PCD partially inhibited electron transport, resulting in oxidative burst in mitochondria. However, ATP production maintained constant. Mitochondria proteome were analyzed by two dimensional IEF/SDS-PAGE before and after 2h-PCD induction, and eight PCD-related proteins were identified. Among them, four proteins were up-regulated after PCD induction, which included glycoside hydrolase, mitochondrial heat shock protein 70, 20S proteasome subunit, and Cu/Zn-superoxide dismutase, and four were down regulated, namely ATP synthase beta subunit, cytochrome c oxidase subunit 6b, S adenosylmethionine synthetase 2, and transcription initiation factor eIF-3 epsilon. These results suggested that ATP synthase may not be the major producer of ATP in mitochondria during the early stage of PCD in rice. Glycoside hydrolase may be involved in ETC impairment and ROS burst, and mitochondrial HSP70 is a potential candidate for PCD regulation. The possible roles of other proteins on PCD initiation were also discussed. PMID- 19217307 TI - A fused silica micro-electrospray tip with an electrically floating metal wire insert to achieve more stable electrospray ionization. AB - A new electrospray tip with a wire insert was tested and compared with the conventional bare fused silica capillary tip. The new tip combined the approach of conventional fused silica spray tips with those containing metal wires. Here, we used a floating wire so that the tips could be prepared and replaced more easily. With the conventional tip, the electrospray process became unstable and the spray current fluctuated significantly in the presence of an air bubble. When the wire-inserted tip was used under the same conditions, much less signal deterioration occurred. The superior performance of this tip over the conventional tip was attributable to its enhanced electric conduction. The new tip has great potential for improving signal stability in LC mass spectrometry. PMID- 19217308 TI - Gas-phase fragmentation of long-lived cysteine radical cations formed via NO loss from protonated S-nitrosocysteine. AB - In this work, we describe two different methods for generating protonated S nitrosocysteine in the gas phase. The first method involves a gas-phase reaction of protonated cysteine with t-butylnitrite, while the second method uses a solution-based transnitrosylation reaction of cysteine with S-nitrosoglutathione followed by transfer of the resulting S-nitrosocysteine into the gas phase by electrospray ionization mass spectrometry (ESI-MS). Independent of the way it was formed, protonated S-nitrosocysteine readily fragments via bond homolysis to form a long-lived radical cation of cysteine (Cys(*+)), which fragments under collision-induced dissociation (CID) conditions via losses in the following relative abundance order: *COOH CH(2)S >> *CH(2)SH approximately = H(2)S. Deuterium labeling experiments were performed to study the mechanisms leading to these pathways. DFT calculations were also used to probe aspects of the fragmentation of protonated S-nitrosocysteine and the radical cation of cysteine. NO loss is found to be the lowest energy channel for the former ion, while the initially formed distonic Cys(*+) with a sulfur radical site undergoes proton and/or H atom transfer reactions that precede the losses of CH(2)S, *COOH, *CH(2)SH, and H(2)S. PMID- 19217309 TI - Metal imaging on surface of micro- and nanoelectronic devices by laser ablation inductively coupled plasma mass spectrometry and possibility to measure at nanometer range. AB - An analytical mass spectrometric method for the elemental analysis of nano bioelectronic devices involved in bioengineering research was developed and applied for measurements of selected metals (Au, Ti, Pt, Cr, etc.) on interdigitated electrode array chips (IDA-chip). An imaging laser ablation inductively coupled plasma mass spectrometric (LA-ICP-MS) procedure was used to map the elements of interest on the surface of the analyzed sample. The obtained images of metals were in a good agreement and corresponded to the micro- and nanofabricated metal electrode pattern. For the analysis at nanometer resolution scale a NF-LA-ICP-MS (NF-near-field) procedure was applied, which utilize thin Ag needle to enhance laser beam energy and improve spatial resolution of the method. The results show a approximately 100x enhancement of analyte signal, when the needle was positioned in the "near-field region" to the sample surface and the laser shot was performed. In addition, mass spectrometric studies of reproducibly for five separated NF-LA shots in different places of analyzed sample yielded an RSD of the measurement of 16%. PMID- 19217310 TI - Sensorimotor specificities in balance control of expert fencers and pistol shooters. AB - Motor skills during sport activity are influenced by practice-related constraints and leads to the development of appropriate postural sensorimotor strategies. Fencing is highly requiring visual monitoring and high-speed motor skills while retaining efficient balance control. Conversely, pistol shooting is a static activity requiring a high control of body sway. This study aimed to evaluate balance control and the related neurosensory organisation through reproducible postural tasks with and without sensory conflict. Twelve expert fencers, 10 expert shooters and 10 sedentary controls have performed a static posturographic test and a sensory organisation test (in 6 different sensory situations based upon sway-referenced vision and support surface, C1 to C6). Shooters yielded a better balance control during C1 (eyes open) and C2 (eyes closed) than fencers and controls. Fencers showed a better balance control in C5 (eyes closed with sway-referenced support surface) than shooters and controls. While this study confirms the beneficial effects of physical activities on balance control, a differential effect on balance characteristics due to the acquired specific motor skills was also noted. In addition to high proprioceptive sensitivity in sportsmen, dynamic constraints in fencing force fencers to permanently select the most relevant information to manage better sensory conflicting situations. PMID- 19217311 TI - Neuromuscular fatigue following high versus low-intensity eccentric exercise of biceps brachii muscle. AB - PURPOSE: This study investigated neuromuscular fatigue following high versus low intensity eccentric exercise corresponding to the same amount of work. METHODS: Ten volunteers performed two eccentric exercises of the elbow flexors: a high intensity versus a low-intensity exercise. Maximal voluntary contraction torque and surface electromyography of the biceps brachii muscle were recorded before, immediately and 48 h after exercises. Maximal voluntary activation level, neural (M-wave) and contractile (muscular twitch) properties of the biceps brachii muscle were analysed using electrical stimulation techniques. RESULTS: Maximal voluntary contraction torque was significantly (P<0.01) reduced immediately and 48 h after exercise but the reduction was not different between the two conditions. Electromyography associated with maximal voluntary contraction significantly decreased (P<0.05) immediately and 48 h after exercise for both conditions while maximal voluntary activation level was only significantly reduced immediately after the high-intensity exercise. Peak twitch alterations were observed immediately and 48 h after exercise for both conditions while M wave did not change. CONCLUSION: High and low-intensity eccentric exercises with the same amount of work induced the same reduction in maximal strength capacities of the biceps brachii muscles. The magnitude of peripheral and central fatigue was very similar in both conditions. PMID- 19217312 TI - Cognitive cost of motor reorganizations associated with muscular fatigue during a repetitive pointing task. AB - Muscular fatigue is known to impair motor performance and to catalyse the development of upper limb musculoskeletal disorders. In order to delay the deleterious effects of muscular fatigue, the central nervous system (CNS) employs compensatory strategies. The cognitive cost of such compensatory strategies was assessed in 10 male subjects who alternatively performed two dual-task protocols before and immediately after an exhaustion procedure specific to upper arm abductor musculature. The main motor tasks were an isometric force-matching and a rapid multi-joint pointing. A secondary probe reaction time (RT) task was performed during both protocols and served as an indicator of attentional demands. Overall motor task performance was maintained despite fatigue. Kinematic and electromyographic data revealed that subjects used motor reorganization during the pointing task when fatigued. The RT increased with fatigue in both dual-task protocols, but this increase was significantly higher during the pointing task than during the force-matching task. The results highlight that the motor reorganization used by the CNS under muscular fatigue states require higher attentional demands than the initial motor organization. Finally, the capacity to delay the deleterious effects of muscular fatigue seems to depend on the proportion of cognitive resources available to plan the compensatory motor strategy. PMID- 19217313 TI - Routine femoral artery pressure monitoring in cardiac surgery. PMID- 19217314 TI - Antibiotic prophylaxis for cardiac surgery: a shift away from traditional cephalosporins? PMID- 19217315 TI - Levosimendan reduces cardiac troponin release after cardiac surgery: a meta analysis of randomized controlled studies. AB - OBJECTIVES: The authors performed a meta-analysis to investigate the effects of levosimendan in cardiac surgery. Inotropic drugs have never shown beneficial effects on outcome in randomized controlled studies, with the possible exception of levosimendan. DESIGN: A meta-analysis. SETTING: Hospitals. PARTICIPANTS: A total of 139 patients from 5 randomized controlled studies were included in the analysis. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Four investigators independently searched BioMedCentral and PubMed. Inclusion criteria were random allocation to treatment, and comparison of levosimendan versus control performed on cardiac surgery patients. Exclusion criteria were duplicate publications, nonhuman experimental studies, and no outcome data. The endpoint was postoperative cardiac troponin release. Levosimendan was associated with a significant reduction in cardiac troponin peak release (weighted mean difference = 2.5 ng/dL [-3.86, -1.14], p = 0.0003) and in time to hospital discharge (weighted mean difference = -1.38 days [-2.78, 0.03], p = 0.05). No other relevant outcome (mortality, myocardial infarction, atrial fibrillation, time on mechanical ventilation, and intensive care unit stay) was improved in those patients receiving levosimendan. CONCLUSIONS: Levosimendan has cardioprotective effects, resulting in reduced postoperative cardiac troponin release. PMID- 19217316 TI - The impact of atrio-biventricular pacing on hemodynamics and left ventricular dyssynchrony compared with atrio-right ventricular pacing alone in the postoperative period after cardiac surgery. AB - OBJECTIVES: The aims of this study were to test the hypotheses that in the postoperative period after coronary artery bypass graft surgery (1) atrio-right ventricular (RA-RV) pacing induces a decrease in cardiac output compared with RA pacing alone and (2) atrio-biventricular (RA-BiV) pacing improves CO compared with RA-RV pacing. DESIGN: A prospective observational study. SETTING: A single center university hospital. PARTICIPANTS: Patients referred for coronary artery bypass graft surgery. INTERVENTIONS: Patients were studied during atrial, RA-RV, and RA-BiV pacing. Cardiac output (echocardiography) and left ventricular dyssynchrony were assessed at each step. MEASUREMENTS AND MAIN RESULTS: RA-RV pacing induced a significant decrease in cardiac output (4.3 +/- 1.0 to 3.7 +/- 0.8 L/min, p < 0.01) and a significant increase in left ventricular dyssynchrony (13 +/- 12 to 80 +/- 25 milliseconds, p < 0.01). Biventricular pacing induced a significant increase in cardiac output (3.7 +/- 0.8 to 4.5 +/- 1.0 L/min, p < 0.01) and a significant decrease in left ventricular dyssynchrony compared with right ventricular pacing (80 +/- 25 to 21 +/- 16 milliseconds, p < 0.05). CONCLUSIONS: RA-BiV pacing improves cardiac output compared with RA-RV pacing in the postoperative period after coronary artery bypass graft surgery. This improvement is related to an improvement in left ventricular synchronicity. PMID- 19217318 TI - In vivo characterization of cytokine profiles and viral load during murine cytomegalovirus-induced acute myocarditis. AB - BACKGROUND: Murine cytomegalovirus (MCMV) is an etiologic agent of acute and chronic myocarditis in BALB/c mice. Immunologic host responses appear to play a key role in pathogenesis but have been incompletely defined. METHODS: BALB/c mice were infected with a sublethal dose of MCMV. Cytokine transcription and viral load (measured by quantitative real-time polymerase chain reaction) and histopathological analyses were performed at specified time points. RESULTS: Increased tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, and interferon (IFN)-gamma, as well as IL-10 mRNA transcripts, were detected in the hearts of infected mice starting at Day 1 post-infection (p.i.), with peak levels occurring at Day 8 p.i. (7-fold, 14-fold, 41-fold, and 16-fold higher than background, respectively). Peak cytokine transcription significantly correlated with a 10 fold increase in viral load (P<.001) at Day 8 p.i. Myocarditis-related pathological changes, measured by infiltration foci, were greatest at Day 8 p.i., corresponding with peak cytokine transcription and significantly correlated with IFN-gamma levels (P<.0001). Infiltration foci were predominantly composed of CD3(+) T cells. Cardiac calcification was observed in most infected mice predominantly over the right ventricle. Histological analysis of heart sections from mice infected with recombinant enhanced green fluorescence protein-MCMV revealed a localized and sporadic pattern of virus throughout all heart layers. CONCLUSIONS: MCMV-induced myocarditis in BALB/c mice is characterized by in vivo production of proinflammatory cytokines in a pattern correlating with MCMV viral load. The infection pattern and inflammatory response is highly localized, sporadic, and involves endocardium, epicardium, as well as the myocardium, with greatest amounts of virus detected in areas of pathologic calcification. PMID- 19217319 TI - The diabetic hand: a forgotten complication? AB - The manifestations of diabetes in the hand were much discussed in the 1970s and 1980s. The present review aims to revisit the diabetic hand and to discuss the pathology of the hand that may be clinically important in diabetic patients. In the strict sense of the term, the "diabetic hand" encompasses the three most widely studied conditions which have traditionally been associated with diabetes, namely limited joint mobility, Dupuytren's contracture and trigger finger. There is evidence that these entities are significantly more frequent in patients with diabetes and also that they may be associated with diabetes duration, poor metabolic control and presence of microvascular complications. In a more general sense, though, there are other conditions affecting the hands, which also occur more frequently in diabetes. From a practical point of view, increased alertness both for neuropathic hand ulcers in patients with profound neuropathy and for diabetic hand infections is absolutely necessary. Recently, reduced hand strength is beginning to be recognized as a further complication of diabetes. Thus, the hand may reveal substantial pathology in diabetes, and ideally, clinical examination should not ignore it. PMID- 19217320 TI - Exendin-4 treatment of nonobese diabetic mice increases beta-cell proliferation and fractional insulin reactive area. AB - OBJECTIVE: The notion of combining immunomodulatory agents with the incretin exendin-4 (Ex-4) has seen considerable favor as a potential therapy for the reversal of type 1 diabetes in man. While the addition of Ex-4 provides modest improvement to the effectiveness of immunological-based monotherapies in reversing hyperglycemia in the nonobese diabetic (NOD) mouse, the mechanism of action underlying this effect remains controversial and formed the basis for this investigation. RESEARCH DESIGN AND METHODS: Female NOD mice with new onset diabetes received either Ex-4 (0.2 microg) or saline via daily intraperitoneal injection for 30 days. To maintain viability after diagnosis of diabetes, animals also received subcutaneous insulin pellets. When persistent hyperglycemia returned, animals were sacrificed and histological studies performed to assess beta-cell proliferation (BrdU+/insulin+; Ki67+/insulin+) and fractional insulin reactive area. RESULTS: Ex-4-treated animals experienced diabetes reversal rates no better than controls. Despite this, Ex-4-treated mice demonstrated increased fractional insulin area (P=.035) and beta-cell proliferation as evidenced by elevated BrdU (P=.0001) and Ki67 staining (P=.04) with insulin co-localization. Also noteworthy, Ex-4-treated mice had poor weight gain following diagnosis in comparison to saline-treated animals (P=.003). CONCLUSIONS: Ex-4 monotherapy (0.2 microg daily-10 microg/kg per day) in NOD mice with new onset diabetes increases beta-cell proliferation and fractional insulin area. Ex-4 remains a promising component of combination therapies for type 1 diabetes. Additional studies are needed to identify a dose that maximizes beta-cell proliferation and minimizes potential side effects. PMID- 19217321 TI - Inflammation induction of Dickkopf-1 mediates chondrocyte apoptosis in osteoarthritic joint. AB - OBJECTIVE: Dysregulated Wnt signaling appears to modulate chondrocyte fate and joint disorders. Dickkopf-1 (DKK1) regulates the pathogenesis of skeletal tissue by inhibiting Wnt actions. This study examined whether DKK1 expression is linked to chondrocyte fate in osteoarthritis (OA). METHOD: Articular cartilage specimens harvested from nine patients with knee OA and from six controls with femoral neck fracture were assessed for DKK1, interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), Bad, Bax, Bcl2 and caspase-3 expression by real time polymerase chain reaction (RT-PCR) and immunohistochemistry. Apoptotic chondrocytes were detected by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labelling (TUNEL) and 4', 6-dianidino-2 phenylindole dihydrochloride (DAPI) staining. Human chondrocyte cultures were treated with recombinant IL-1beta and monoclonal DKK1 antibody to determine whether DKK1 impairs chondrocyte survival. RESULTS: Expression of DKK1 correlated with inflammatory cytokine levels (IL-1beta and TNF-alpha expressions), proapoptosis regulators (Bad and caspase-3 expressions) and TUNEL staining in OA cartilage tissues. The IL-1beta induced expressions of DKK1, Bax, Bad and caspase 3-dependent apoptosis of chondrocyte cultures. Neutralization of DKK1 by monoclonal DKK1 antibody significantly abrogated IL-1beta-mediated caspase-3 cleavage and apoptosis and reversed chondrocyte proliferation. Recombinant DKK1 treatment impaired chondrocyte growth and promoted apoptosis. By suppressing nuclear beta-catenin accumulation and Akt phosphorylation, DKK1 mediated IL-1beta promotion of chondrocyte apoptosis. CONCLUSION: Chondrocyte apoptosis correlates with joint OA. Expression of DKK1 contributes to cartilage deterioration and is a potent factor in OA pathogenesis. Attenuating DKK1 may reduce cartilage deterioration in OA. PMID- 19217322 TI - Relative efficacies of omega-3 polyunsaturated fatty acids in reducing expression of key proteins in a model system for studying osteoarthritis. AB - OBJECTIVE: To assess the relative efficacy of three different omega-3 (n-3) polyunsaturated fatty acids (PUFAs) in suppressing the mRNA levels for important proteins involved in the etiology of osteoarthritis (OA). METHODS: A model cell culture system (bovine chondrocytes) was used. Inflammatory factors and enzymes involved in OA were induced by exposure of the chondrocyte cultures to interleukin-1alpha (IL-1alpha). The effect of pre-incubating cultures with various amounts of exogenous fatty acids on subsequent levels of mRNAs was assessed by reverse transcription-polymerase chain reactions (RT-PCR). RESULTS: Exposure of cultures to IL-1alpha induced expression of the cartilage proteinases A Disintegrin And Metalloproteinase with ThromboSpondin motifs (ADAMTS)-4 and ADAMTS-5, cyclooxygenase (COX)-2, the matrix metalloproteinase (MMP)-3 and the inflammatory cytokines IL-1alpha, interleukin-1beta (IL-1beta) and tumour necrosis factor-alpha (TNF-alpha). n-3 PUFAs were able to reduce the levels of mRNA for ADAMTS-4, ADAMTS-5, MMP-3, MMP-13, COX-2 (but not COX-1), IL-1alpha, IL 1beta and TNF-alpha. Eicosapentaenoic acid (EPA) was the most effective, followed by docosahexaenoic (DHA) and then alpha-linolenic (ALA) acid. The n-6 PUFA, arachidonic acid (AA) had no effect. CONCLUSION: These results show that omega-3 (n-3) PUFAs cause a reduction in the mRNA levels for various proteins known to be important in the pathology of OA. They provide a molecular explanation, at least in part, for beneficial effects of dietary omega-3 PUFAs for the amelioration of symptoms of the disease. The relative efficacy of EPA suggests that this omega-3 PUFA may be especially useful for dietary supplementation in patients with OA. PMID- 19217323 TI - NKT cell-plasmacytoid dendritic cell cooperation via OX40 controls viral infection in a tissue-specific manner. AB - Invariant natural killer T (iNKT) cells promote immune responses to various pathogens, but exactly how iNKT cells control antiviral responses is unclear. Here, we showed that iNKT cells induced tissue-specific antiviral effects in mice infected by lymphocytic choriomeningitis virus (LCMV). Indeed, iNKT cells inhibited viral replication in the pancreas and liver but not in the spleen. In the pancreas, iNKT cells expressed the OX40 molecule and promoted type I interferon (IFN) production by plasmacytoid dendritic cells (pDCs) through OX40 OX40 ligand interaction. Subsequently, this iNKT cell-pDC cooperation attenuated the antiviral adaptive immune response in the pancreas but not in the spleen. The dampening of pancreatic anti-LCMV CD8(+) T cell response prevented tissue damage in transgenic mice expressing LCMV protein in islet beta cells. Thus, this study identifies pDCs as an essential partner of iNKT cells for mounting an efficient, nondeleterious antiviral response in peripheral tissue. PMID- 19217324 TI - Roquin differentiates the specialized functions of duplicated T cell costimulatory receptor genes CD28 and ICOS. AB - During evolutionary adaptation in the immune system, host defense is traded off against autoreactivity. Signals through the costimulatory receptor CD28 enable T cells to respond specifically to pathogens, whereas those through the related costimulatory receptor, ICOS, which arose by gene duplication, are critical for affinity maturation and memory antibody responses. ICOS ligand, unlike the pathogen-inducible CD28 ligands, is widely and constitutively expressed in the immune system. Here, we show that crosstalk between these two pathways provides a mechanism for obviating the normal T cell dependence on CD28. Several CD28 mediated responses-generation of follicular helper T cells, germinal center formation, T helper 1 cell-dependent extrafollicular antibody responses to Salmonella and bacterial clearance, and regulatory T cell homeostasis-became independent of CD28 and dependent on ICOS when the E3 ubiquitin ligase Roquin was mutated. Mechanisms to functionally compartmentalize ICOS and CD28 signals are thus critical for two-signal control of normal immune reactions. PMID- 19217325 TI - Lunatic and manic fringe cooperatively enhance marginal zone B cell precursor competition for delta-like 1 in splenic endothelial niches. AB - Notch2 activation induced by Delta-like-1 (DL1) drives development of splenic marginal zone (MZ) B cells, an innate-like lineage that protects against sepsis. DL1 interacts with Notch2 weakly, but it is not known whether enhancement of DL1 induced Notch2 activation by Fringe glycosyltransferases is important for MZ B cell development. Furthermore, DL1-expressing cells that promote MZ B cell development have not been identified. We show that Lunatic Fringe (Lfng) and Manic Fringe (Mfng) cooperatively enhanced the DL1-Notch2 interaction to promote MZ B cell development. We also identified radio-resistant red pulp endothelial cells in the splenic MZ that express high amounts of DL1 and promoted MZ B generation. Finally, MZ B cell precursor competition for DL1 homeostatically regulated entry into the MZ B cell pool. Our study has revealed that the Fringe Notch2 interaction has important functions in vivo and provides insights into mechanisms regulating MZ B cell development. PMID- 19217326 TI - Increased metabolic activity in abdominal aortic aneurysm detected by 18F fluorodeoxyglucose (18F-FDG) positron emission tomography/computed tomography (PET/CT). AB - OBJECTIVES: Abdominal aortic aneurysms (AAAs) are associated with an inflammatory cell infiltrate and enzymatic degradation of the vessel wall. The aim of this study was to detect increased metabolic activity in the wall of the AAA with 18F fluorodeoxyglucose ((18)F-FDG), mediated by glucose transporter protein (GLUTs), using a dedicated hybrid PET/64-detector CT. DESIGN, METHOD AND MATERIALS: 14 patients (All male, mean age 73.6 years, range 61-82) with AAA under surveillance underwent PET/CT scanning with 175 MBq of intravenous (18)F-FDG. The maximum aneurysm diameter and calcification score were determined on the attenuation correction CT. A volume of interest was placed on the aneurysm sac and the maximum Standardised Uptake Value (SUV(max)) measured. RESULTS: The mean aneurysm diameter was 5.4 cm (SD+/-0.8). Two aneurysms had the CT characteristics of inflammatory aneurysms. Twelve aneurysms showed increased FDG uptake (SUV(max)>2.5). There was no significant difference in FDG uptake between heavily calcified aneurysms and non-heavily calcified aneurysms (t-test). There was a significant increase in the FDG uptake in the two inflammatory aneurysms compared to the other twelve aneurysms (t-test; P=0.04). CONCLUSION: The findings in this study offer in vivo evidence that the AAA wall shows increased glucose metabolism, mediated by the GLUTs: this increased metabolic activity as detected by PET/CT may be present in most AAAs. PMID- 19217327 TI - Re: Is endodontic treatment passe? PMID- 19217328 TI - Facial skin sensibility in a young healthy chinese population. AB - OBJECTIVE: To quantify normal neurosensory facial sensibility in a young healthy Chinese population for use as a reference when evaluating postoperative nerve damage. STUDY DESIGN: One hundred consecutive eligible normal young Chinese individuals were included. Each subject underwent objective neurosensory testing (static light touch, 2-point static, and pain detection thresholds) at 8 facial sites within the distribution of the trigeminal nerve. Data were calculated into means and standard deviations, and paired t tests were used to compare values between the left and right sides and quadrants; unpaired t test was used to compare the values between genders. A P value of 50 cc with those with prostate volumes 50 cc (mean 58.1 cc, range 50.2-86.0 cc); the mean D(90), V(100), V(150), and V(200) was 125.1%, 95.2%, 68.2%, and 41.7%, respectively. The rectal V(100) was 1.0 cc for both cohorts. There was no statistically significant difference between the cohorts with respect to postimplant dosimetry for D(90), V(100), and V(150). The V(200) for prostate volumes >50 cc was significantly lower (p<0.05). CONCLUSIONS: In the present study, patients with prostate volumes >50 cc have postimplant dosimetry parameters similar to patients with prostate volumes or=15%/y was defined as clinically significant progression. The relationship between race and CAC progression was evaluated with multivariable linear and logistic regression models controlling for age and other cardiovascular risk factors. RESULTS: African Americans had significantly lower baseline CAC scores (34.3 vs 101.5; P = 0.004); lower follow-up CAC scores (56.6 vs 180.6; P = 0.001); and worse cardiovascular risk profiles. The annualized CAC progression rate was not significantly related to race in the multivariable linear regression model controlling for age, the Framingham risk score, and other cardiovascular risk factors. Significant CAC progression occurred in 43.5% of all participants. The incidence of significant progression of CAC for African American and white men was similar (53.1% vs 52.4%; P = 0.94), even when controlling for age, the Framingham risk score, and other cardiovascular risk factors. CONCLUSION: Although African American men have less CAC than white men, CAC progression occurs at a comparable rate over 4 years. PMID- 19217368 TI - The curious case of a wandering kinase: CaMKII spreads the wealth? AB - Calcium/calmodulin-dependent kinase II has been suggested to produce input specific long-term potentiation of synaptic strength. This idea has been complicated by results from Rose, Jin, and Craig demonstrating that spatiotemporally restricted NMDA receptor excitation at contiguous synapses can result in the translocation of activated CaMKII throughout the dendritic arbor. PMID- 19217369 TI - Synaptic vesicle endocytosis: get two for the price of one? AB - Tight coupling between synaptic vesicle exocytosis and endocytosis is critical for the maintenance of neurotransmission. In this issue of Neuron, Zhu et al. reveal a surprising facet of this coupling by showing that, at low frequencies, fusion of a single vesicle leads to retrieval of two vesicles with dissimilar attributes. PMID- 19217370 TI - Hooked on the D3 receptor: CaMKII's new addiction. AB - The dopaminergic D3 receptor (D3R) is emerging as an antagonist of sensitized behavior induced by addictive drugs such as cocaine. New work by Wang and coworkers in this issue of Neuron now identifies CaMKII as an important negative regulator of the D3R. CaMKII relieves the D3R-mediated inhibition on sensitized behavior to foster drug-seeking behavior. PMID- 19217371 TI - Reprogramming therapeutics: iPS cell prospects for neurodegenerative disease. AB - The recent description of somatic cell reprogramming to an embryonic stem (ES) cell-like phenotype, termed induced pluripotent stem (iPS) cell technology, presents an exciting potential venue toward cell-based therapeutics and disease models for neurodegenerative disorders. Two recent studies (Dimos et al. and Ebert et al.) describe the initial characterization of neurodegenerative disease patient-derived iPS cell cultures as proof of concept for the utility of this technology. PMID- 19217373 TI - Heterosynaptic molecular dynamics: locally induced propagating synaptic accumulation of CaM kinase II. AB - Calcium-calmodulin-dependent protein kinase II (CaMKII) is a key mediator of synaptic plasticity and learning. Global pyramidal cell glutamate stimulation induces translocation of CaMKII from dendritic shafts to spines. Here we show that local dendritic stimulation by puffing glutamate onto a region containing 7 32 synapses induces translocation of CaMKII to synapses initially at the puff site but that translocation subsequently spreads within dendrites to the distal dendrite arbor, resulting in a persistent, widespread synaptic accumulation. This locally induced propagating synaptic (L-IPS) accumulation of CaMKII requires activation of NMDA receptors and L-type Ca(2+) channels and is preceded by a Ca(2+) spike. L-IPS translocation of CaMKII alters biochemical signaling and is associated with an increase in AMPA receptor GluR1 at both stimulated and nonstimulated synapses and thus provides a molecular mechanism for heterosynaptic plasticity. PMID- 19217372 TI - Synaptic AMPA receptor plasticity and behavior. AB - The ability to change behavior likely depends on the selective strengthening and weakening of brain synapses. The cellular models of synaptic plasticity, long term potentiation (LTP) and depression (LTD) of synaptic strength, can be expressed by the synaptic insertion or removal of AMPA receptors (AMPARs), respectively. We here present an overview of studies that have used animal models to show that such AMPAR trafficking underlies several experience-driven phenomena from neuronal circuit formation to the modification of behavior. We argue that monitoring and manipulating synaptic AMPAR trafficking represents an attractive means to study cognitive function and dysfunction in animal models. PMID- 19217374 TI - FARP1 promotes the dendritic growth of spinal motor neuron subtypes through transmembrane Semaphorin6A and PlexinA4 signaling. AB - The dendritic morphology of neurons dictates their abilities to process and transmit information; however, the signaling pathways that regulate dendritic growth and complexity are poorly understood. Here, we show that retinoids induce the expression of the FERM Rho-GEF protein FARP1 in the developing spinal cord. FARP1 is expressed in subsets of motor neurons and is enriched in dendrites of lateral motor column (LMC) neurons that innervate the limb. FARP1 is necessary and sufficient to promote LMC dendritic growth but does not affect dendrite number or axonal morphology. We show that FARP1 serves as a specific effector of transmembrane Semaphorin6A and PlexinA4 signals to regulate LMC dendritic growth, and that its Rho-GEF domain is necessary for this function. These findings reveal that retinoid and Sema6A/PlexA4 signaling pathways intersect through FARP1 to control dendritic growth, and uncover the existence of subtype-specific signaling networks that control dendritic developmental programs in spinal motor neurons. PMID- 19217375 TI - Motor control in a Drosophila taste circuit. AB - Tastes elicit innate behaviors critical for directing animals to ingest nutritious substances and reject toxic compounds, but the neural basis of these behaviors is not understood. Here, we use a neural silencing screen to identify neurons required for a simple Drosophila taste behavior and characterize a neural population that controls a specific subprogram of this behavior. By silencing and activating subsets of the defined cell population, we identify the neurons involved in the taste behavior as a pair of motor neurons located in the subesophageal ganglion (SOG). The motor neurons are activated by sugar stimulation of gustatory neurons and inhibited by bitter compounds; however, experiments utilizing split-GFP detect no direct connections between the motor neurons and primary sensory neurons, indicating that further study will be necessary to elucidate the circuitry bridging these populations. Combined, these results provide a general strategy and a valuable starting point for future taste circuit analysis. PMID- 19217377 TI - Two pathways of synaptic vesicle retrieval revealed by single-vesicle imaging. AB - Synaptic vesicle recycling is essential for maintaining efficient synaptic transmission. Detailed dissection of single-vesicle recycling still remains a major challenge. We have developed a fluorescent pH reporter that permits us to follow the fate of individual vesicles at hippocampal synapses after exocytosis. Here we show that, during low-frequency stimulation, single-vesicle fusion leads to two distinct vesicle internalizations, instead of one, as in general perception: one by a fast endocytosis pathway ( approximately 3 s), the other by a slow endocytosis pathway (after 10 s). The exocytosed vesicular proteins are preferentially recaptured in both pathways. RNAi knockdown of clathrin inhibits both pathways. As stimulation frequency increases, the number of endocytosed vesicles begins to match antecedent exocytosis. Meanwhile, the slow endocytosis is accelerated and becomes the predominant pathway. These results reveal that two pathways of endocytosis are orchestrated during neuronal activity, establishing a highly efficient endocytosis at central synapses. PMID- 19217376 TI - A transmembrane accessory subunit that modulates kainate-type glutamate receptors. AB - Glutamate receptors play major roles in excitatory transmission in the vertebrate brain. Among ionotropic glutamate receptors (AMPA, kainate, NMDA), AMPA receptors mediate fast synaptic transmission and require TARP auxiliary subunits. NMDA receptors and kainate receptors play roles in synaptic transmission, but it remains uncertain whether these ionotropic glutamate receptors also have essential subunits. Using a proteomic screen, we have identified NETO2, a brain specific protein of unknown function, as an interactor with kainate-type glutamate receptors. NETO2 modulates the channel properties of recombinant and native kainate receptors without affecting trafficking of the receptors and also modulates kainate-receptor-mediated mEPSCs. Furthermore, we found that kainate receptors regulate the surface expression of NETO2 and that NETO2 protein levels and surface expression are decreased in mice lacking the kainate receptor GluR6. The results show that NETO2 is a kainate receptor subunit with significant effects on glutamate signaling mechanisms in brain. PMID- 19217378 TI - Snapin facilitates the synchronization of synaptic vesicle fusion. AB - Synaptic vesicle (SV) fusion is a fine-tuned process requiring a concert of fusion machineries. Using cortical neurons from snapin-deficient mice, we reveal a role for Snapin in facilitating synchronous release. In addition to reduced frequency of miniature excitatory postsynaptic currents (mini-EPSCs) and smaller release-ready vesicle pool (RRP) size, snapin deficiency results in EPSCs with multiple peaks and increased rise and decay times, reflecting "desynchronized" SV fusion. These defects impair both synaptic precision and efficacy during sustained neurotransmission. Transient expression of Snapin not only rescues the slowed kinetics of EPSCs, but also further accelerates the rate found in wild type neurons. Furthermore, expression of Snapin-C66A, a dimerization-defective mutant with impaired interactions with SNAP-25 and Synaptotagmin, reduces the RRP size but exhibits less effect on synchronized fusion. Our studies provide mechanistic insights into a dual role of Snapin in enhancing the efficacy of SV priming and in fine-tuning synchronous SV fusion. PMID- 19217379 TI - Activity-dependent modulation of limbic dopamine D3 receptors by CaMKII. AB - Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) is central to synaptic transmission. Here we show that synaptic CaMKIIalpha binds to the N-terminal region of the third intracellular loop of the limbic dopamine D3 receptor (D3R). This binding is Ca(2+) sensitive and is sustained by autophosphorylation of CaMKII, providing an unrecognized route for the Ca(2+)-mediated regulation of D3Rs. The interaction of CaMKIIalpha with D3Rs transforms D3Rs into a biochemical substrate of the kinase and promotes the kinase to phosphorylate D3Rs at a selective serine site (S229). In accumbal neurons in vivo, CaMKIIalpha is recruited to D3Rs by rising Ca(2+) to increase the CaMKIIalpha-mediated phosphorylation of D3Rs, thereby transiently inhibiting D3R efficacy. Notably, the D3R inhibition is critical for integrating dopamine signaling to control behavioral sensitivity to the psychostimulant cocaine. Our data identify CaMKIIalpha as a recruitable regulator of dopamine receptor function. By binding and phosphorylating limbic D3Rs, CaMKIIalpha modulates dopamine signaling and psychomotor function in an activity-dependent manner. PMID- 19217380 TI - Robust microcircuit synchronization by inhibitory connections. AB - Microcircuits in different brain areas share similar architectural and biophysical properties with compact motor networks known as central pattern generators (CPGs). Consequently, CPGs have been suggested as valuable biological models for understanding of microcircuit dynamics and particularly, their synchronization. We use a well known compact motor network, the lobster pyloric CPG to study principles of intercircuit synchronization. We couple separate pyloric circuits obtained from two animals via artificial synapses and observe how their synchronization depends on the topology and kinetic parameters of the computer-generated synapses. Stable in-phase synchronization appears when electrically coupling the pacemaker groups of the two networks, but reciprocal inhibitory connections produce more robust and regular cooperative activity. Contralateral inhibitory connections offer effective synchronization and flexible setting of the burst phases of the interacting networks. We also show that a conductance-based mathematical model of the coupled circuits correctly reproduces the observed dynamics illustrating the generality of the phenomena. PMID- 19217381 TI - Mechanisms of sleep-dependent consolidation of cortical plasticity. AB - Sleep is thought to consolidate changes in synaptic strength, but the underlying mechanisms are unknown. We investigated the cellular events involved in this process during ocular dominance plasticity (ODP)-a canonical form of in vivo cortical plasticity triggered by monocular deprivation (MD) and consolidated by sleep via undetermined, activity-dependent mechanisms. We find that sleep consolidates ODP primarily by strengthening cortical responses to nondeprived eye stimulation. Consolidation is inhibited by reversible, intracortical antagonism of NMDA receptors (NMDARs) or cAMP-dependent protein kinase (PKA) during post-MD sleep. Consolidation is also associated with sleep-dependent increases in the activity of remodeling neurons and in the phosphorylation of proteins required for potentiation of glutamatergic synapses. These findings demonstrate that synaptic strengthening via NMDAR and PKA activity is a key step in sleep dependent consolidation of ODP. PMID- 19217382 TI - Task difficulty and performance induce diverse adaptive patterns in gain and shape of primary auditory cortical receptive fields. AB - Attention is essential for navigating complex acoustic scenes, when the listener seeks to extract a foreground source while suppressing background acoustic clutter. This study explored the neural correlates of this perceptual ability by measuring rapid changes of spectrotemporal receptive fields (STRFs) in primary auditory cortex during detection of a target tone embedded in noise. Compared with responses in the passive state, STRF gain decreased during task performance in most cells. By contrast, STRF shape changes were excitatory and specific, and were strongest in cells with best frequencies near the target tone. The net effect of these adaptations was to accentuate the representation of the target tone relative to the noise by enhancing responses of near-target cells to the tone during high-signal-to-noise ratio (SNR) tasks while suppressing responses of far-from-target cells to the masking noise in low-SNR tasks. These adaptive STRF changes were largest in high-performance sessions, confirming a close correlation with behavior. PMID- 19217383 TI - Gambling near-misses enhance motivation to gamble and recruit win-related brain circuitry. AB - "Near-miss" events, where unsuccessful outcomes are proximal to the jackpot, increase gambling propensity and may be associated with the addictiveness of gambling, but little is known about the neurocognitive mechanisms that underlie their potency. Using a simplified slot machine task, we measured behavioral and neural responses to gambling outcomes. Compared to "full-misses," near-misses were experienced as less pleasant, but increased desire to play. This effect was restricted to trials where the subject had personal control over arranging their gamble. Near-miss outcomes recruited striatal and insula circuitry that also responded to monetary wins; in addition, near-miss-related activity in the rostral anterior cingulate cortex varied as a function of personal control. Insula activity to near-misses correlated with self-report ratings as well as a questionnaire measure of gambling propensity. These data indicate that near misses invigorate gambling through the anomalous recruitment of reward circuitry, despite the objective lack of monetary reinforcement on these trials. PMID- 19217384 TI - Entry of herpesviruses into cells: more than one way to pull the trigger. AB - Interaction of viral protein gp42 with its receptor is the trigger for the entry of Epstein-Barr virus into B cells. The structure of gp42 reported by Kirschner etal. (2009) in this issue of Structure suggests a likely triggering mechanism substantially different from that of a related herpesvirus. PMID- 19217385 TI - For structural biology, try infrared instead. AB - 1D and 2D-IR spectroscopy was used to resolve the structural transition that is responsible for the gating mechanism of the influenza A M2 channel (Manor et al., 2009). This report constitutes a milestone in the development of both ATR-FTIR and 2D-IR spectroscopies as precise tools for structural biology. PMID- 19217387 TI - The nucleosome family: dynamic and growing. AB - Ever since the discovery of the nucleosome in 1974, scientists have stumbled upon discrete particles in which DNA is wrapped around histone complexes of different stoichiometries: octasomes, hexasomes, tetrasomes, "split" half-nucleosomes, and, recently, bona fide hemisomes. Do all these particles exist in vivo? Under what conditions? What is their physiological significance in the complex DNA transactions in the eukaryotic nucleus? What are their dynamics? This review summarizes research spanning more than three decades and provides a new meaning to the term "nucleosome." The nucleosome can no longer be viewed as a single static entity: rather, it is a family of particles differing in their structural and dynamic properties, leading to different functionalities. PMID- 19217386 TI - Outcome of a workshop on applications of protein models in biomedical research. AB - We describe the proceedings and conclusions from the "Workshop on Applications of Protein Models in Biomedical Research" (the Workshop) that was held at the University of California, San Francisco on 11 and 12 July, 2008. At the Workshop, international scientists involved with structure modeling explored (i) how models are currently used in biomedical research, (ii) the requirements and challenges for different applications, and (iii) how the interaction between the computational and experimental research communities could be strengthened to advance the field. PMID- 19217388 TI - Wheel of Life, Wheel of Death: A Mechanistic Insight into Signaling by STAND Proteins. AB - The signal transduction ATPases with numerous domains (STAND) represent a newly recognized class of widespread, sophisticated ATPases that are related to the AAA+ proteins and that function as signaling hubs. These proteins control diverse biological processes in bacteria and eukaryotes, including gene expression, apoptosis, and innate immunity responses. They function as tightly regulated switches, with the off and on positions corresponding to a long-lived monomeric, ADP-bound form and a multimeric, ATP-bound form, respectively. Inducer binding to the sensor domain activates the protein by promoting ADP for ATP exchange, probably through removal of an intramolecular inhibitory interaction, whereas ATP hydrolysis turns off the protein. One key component of the switch is a three domain module carrying the ATPase activity (nucleotide-binding oligomerization domain [NOD]). Analysis of the atomic structures of four crystallized nucleotide bound NOD modules provides an unprecedented insight into the NOD conformational changes underlying the activation process. PMID- 19217389 TI - "Conditional Restraints": Restraining the Free Atoms in ARP/wARP. AB - The automated building of a protein model into an electron density map remains a challenging problem. In the ARP/wARP approach, model building is facilitated by initially interpreting a density map with free atoms of unknown chemical identity; all structural information for such chemically unassigned atoms is discarded. Here, this is remedied by applying restraints between free atoms, and between free atoms and a partial protein model. These are based on geometric considerations of protein structure and tentative (conditional) assignments for the free atoms. Restraints are applied in the REFMAC5 refinement program and are generated on an ad hoc basis, allowing them to fluctuate from step to step. A large set of experimentally phased and molecular replacement structures showcases individual structures where automated building is improved drastically by the conditional restraints. The concept and implementation we present can also find application in restraining geometries, such as hydrogen bonds, in low-resolution refinement. PMID- 19217390 TI - Structural analysis of ligand stimulation of the histidine kinase NarX. AB - Histidine kinase receptors are a large family of membrane-spanning proteins found in many prokaryotes and some eukaryotes. They are a part of two-component signal transduction systems, which each comprise a sensor kinase and a response regulator and are involved with the regulation of many cellular processes. NarX is a histidine kinase receptor that responds to nitrate and nitrite to effect regulation of anaerobic respiration in various bacteria. We present high resolution X-ray crystal structures of the periplasmic sensor domain from Escherichia coli NarX in a complex with nitrate and in the apo state. Our analysis reveals that nitrate-binding induces conformation changes that result in a piston-type displacement between the N- and C-terminal helices of the periplasmic domain. Such conformational changes might represent a conserved mechanism of signaling in histidine kinases by which ligand binding is communicated across the lipid bilayer. PMID- 19217391 TI - Structural basis for p300 Taz2-p53 TAD1 binding and modulation by phosphorylation. AB - Coactivators CREB-binding protein and p300 play important roles in mediating the transcriptional activity of p53. Until now, however, no detailed structural information has been available on how any of the domains of p300 interact with p53. Here, we report the NMR structure of the complex of the Taz2 (C/H3) domain of p300 and the N-terminal transactivation domain of p53. In the complex, p53 forms a short alpha helix and interacts with the Taz2 domain through an extended surface. Mutational analyses demonstrate the importance of hydrophobic residues for complex stabilization. Additionally, they suggest that the increased affinity of Taz2 for p53(1-39) phosphorylated at Thr(18) is due in part to electrostatic interactions of the phosphate with neighboring arginine residues in Taz2. Thermodynamic experiments revealed the importance of hydrophobic interactions in the complex of Taz2 with p53 phosphorylated at Ser(15) and Thr(18). PMID- 19217392 TI - Insights into the architecture of the replicative helicase from the structure of an archaeal MCM homolog. AB - The minichromosome maintenance (MCM) proteins, members of the AAA+ (ATPase associated with diverse cellular activities) superfamily, are believed to constitute the replicative helicase in eukaryotic and archaeal species. Here, we present the 1.9 A resolution crystal structure of a monomeric MCM homolog from Methanopyrus kandleri, the first crystallographic structure of a full-length MCM. We also present an 18 A cryo-electron microscopy reconstruction of the hexameric MCM from Methanothermobacter thermautotrophicus, and fit the atomic resolution crystal structure into the reconstruction in order to generate an atomic model for the oligomeric assembly. These structural data reveal a distinct active site topology consisting of a unique arrangement of critical determinants. The structures also provide a molecular framework for understanding the functional contributions of trans-acting elements that facilitate intersubunit crosstalk in response to DNA binding and ATP hydrolysis. PMID- 19217393 TI - Structure of Epstein-Barr virus glycoprotein 42 suggests a mechanism for triggering receptor-activated virus entry. AB - Epstein-Barr virus requires glycoproteins gH/gL, gB, and gp42 to fuse its lipid envelope with B cells. Gp42 is a type II membrane protein consisting of a flexible N-terminal region, which binds gH/gL, and a C-terminal lectin-like domain that binds to the B-cell entry receptor human leukocyte antigen (HLA) class II. Gp42 triggers membrane fusion after HLA binding, a process that requires simultaneous binding to gH/gL and a functional hydrophobic pocket in the lectin domain adjacent to the HLA binding site. Here we present the structure of gp42 in its unbound form. Comparisons to the previously determined structure of a gp42:HLA complex reveals additional N-terminal residues forming part of the gH/gL binding site and structural changes in the receptor binding domain. Although the core of the lectin domain remains similar, significant shifts in two loops and an alpha helix bordering the essential hydrophobic pocket suggest a structural mechanism for triggering fusion. PMID- 19217394 TI - Comprehensive structural classification of ligand-binding motifs in proteins. AB - Comprehensive knowledge of protein-ligand interactions should provide a useful basis for annotating protein functions, studying protein evolution, engineering enzymatic activity, and designing drugs. To investigate the diversity and universality of ligand-binding sites in protein structures, we conducted the all against-all atomic-level structural comparison of over 180,000 ligand-binding sites found in all the known structures in the Protein Data Bank by using a recently developed database search and alignment algorithm. By applying a hybrid top-down-bottom-up clustering analysis to the comparison results, we determined approximately 3000 well-defined structural motifs of ligand-binding sites. Apart from a handful of exceptions, most structural motifs were found to be confined within single families or superfamilies, and to be associated with particular ligands. Furthermore, we analyzed the components of the similarity network and enumerated more than 4000 pairs of structural motifs that were shared across different protein folds. PMID- 19217395 TI - Gating mechanism of the influenza A M2 channel revealed by 1D and 2D IR spectroscopies. AB - The pH-controlled M2 protein from influenza A is a critical component of the virus and serves as a target for the aminoadamantane antiflu agents that block its H+ channel activity. To better understand its H+ gating mechanism, we investigated M2 in lipid bilayers with a new combination of IR spectroscopies and theory. Linear Fourier transform infrared (FTIR) spectroscopy was used to measure the precise orientation of the backbone carbonyl groups, and 2D infrared (IR) spectroscopy was used to identify channel-lining residues. At low pH (open state), our results match previously published solid-state NMR and X-ray structures remarkably well. However, at neutral pH when the channel is closed, our measurements indicate that a large conformational change occurs that is consistent with the transmembrane alpha-helices rotating by one amino acid register--a structural rearrangement not previously observed. The combination of simulations and isotope-labeled FTIR and 2D IR spectroscopies provides a noninvasive means of interrogating the structures of membrane proteins in general and ion channels in particular. PMID- 19217396 TI - Crystal structure of the N-terminal domain of the secretin GspD from ETEC determined with the assistance of a nanobody. AB - Secretins are among the largest bacterial outer membrane proteins known. Here we report the crystal structure of the periplasmic N-terminal domain of GspD (peri GspD) from the type 2 secretion system (T2SS) secretin in complex with a nanobody, the VHH domain of a heavy-chain camelid antibody. Two different crystal forms contained the same compact peri-GspD:nanobody heterotetramer. The nanobody contacts peri-GspD mainly via CDR3 and framework residues. The peri-GspD structure reveals three subdomains, with the second and third subdomains exhibiting the KH fold which also occurs in ring-forming proteins of the type 3 secretion system. The first subdomain of GspD is related to domains in phage tail proteins and outer membrane TonB-dependent receptors. A dodecameric peri-GspD model is proposed in which a solvent-accessible beta strand of the first subdomain interacts with secreted proteins and/or T2SS partner proteins by beta strand complementation. PMID- 19217397 TI - Reconstruction of the P2X(2) receptor reveals a vase-shaped structure with lateral tunnels above the membrane. AB - In response to the intercellular messenger ATP, P2X receptors transfer various sensory information, including pain. Here we have reconstructed the structure of the P2X(2) receptor at 15 A resolution from more than 90,000 particle images, taken with a cryo-electron microscope equipped with a helium-cooled stage. This three-dimensional depiction, presumably in a closed state, revealed an elongated vase-shaped structure 202 A in height and 160 A in major diameter. The extracellular and transmembrane domains present a two-layered structure, in which a sparse outer layer surrounds a pore-forming inner density. The decreased diameter of a putative ion-conducting pathway at the middle of the membrane was considered to be the narrowest part of the pore, which has been predicted from electrophysiological studies. The sparse, extended structure of the P2X(2) receptor indicates a loose assembly of subunits, which could be a basis for the activation-dependent pore dilation of P2X receptors. PMID- 19217398 TI - Structural basis of m(7)GpppG binding to poly(A)-specific ribonuclease. AB - Poly(A)-specific ribonuclease (PARN) is a homodimeric, processive, and cap interacting 3' exoribonuclease that efficiently degrades eukaryotic mRNA poly(A) tails. The crystal structure of a C-terminally truncated PARN in complex with m(7)GpppG reveals that, in one subunit, m(7)GpppG binds to a cavity formed by the RRM domain and the nuclease domain, whereas in the other subunit, it binds almost exclusively to the RRM domain. Importantly, our structural and competition data show that the cap-binding site overlaps with the active site in the nuclease domain. Mutational analysis demonstrates that residues involved in m(7)G recognition are crucial for cap-stimulated deadenylation activity, and those involved in both cap and poly(A) binding are important for catalysis. A modeled PARN, which shows that the RRM domain from one subunit and the R3H domain from the other subunit enclose the active site, provides a structural foundation for further studies to elucidate the mechanism of PARN-mediated deadenylation. PMID- 19217399 TI - Cryo-EM structure of the native GroEL-GroES complex from thermus thermophilus encapsulating substrate inside the cavity. AB - The chaperonin GroEL interacts with various proteins, leading them to adopt their correct conformations with the aid of GroES and ATP. The actual mechanism is still being debated. In this study, by use of cryo-electron microscopy, we determined the solution structure of the Thermus thermophilus GroEL-GroES complex encapsulating its substrate proteins. We observed the averaged density of substrate proteins in the center of the GroEL-GroES cavity. The position of the averaged substrate density in the cavity suggested a repulsive interaction between a majority of the substrate proteins and the interior wall of the cavity, which is suitable for substrate release. In addition, we observed a distortion of the cis-GroEL ring, especially at the position near the substrate, which indicated that the interaction between the encapsulated proteins and the GroEL ring results in an adjustment in the cavity's shape to accommodate the substrate. PMID- 19217400 TI - Solution structure of the U11-48K CHHC zinc-finger domain that specifically binds the 5' splice site of U12-type introns. AB - The formation of stable 18S U11/U12 di-snRNPs before their association with the pre-mRNA is a characteristic feature of the minor spliceosome. During the spliceosomal assembly, the 18S snRNP binds cooperatively to the introns' 5' splice and branch point site. The molecular basis for this recognition is still unknown. Here, we report the solution structure of the U11-48K CHHC Zn finger, a domain unique to the minor spliceosome. The CHHC Zn-finger structure revealed an unexpected similarity to the TFIIIA domains, with distinct features originating from the type and separation of the zinc-coordinating residues. We show that this domain specifically binds the 5' splice site sequence of U12-type introns when base paired to U11 snRNA in vitro and hence may contribute to the U12 intron recognition. We propose a model in which the U11-48K Zn finger stabilizes U11-5' splice site base pairing and thus plays an important role during the minor spliceosome assembly. PMID- 19217402 TI - Nonproteolytic functions of ubiquitin in cell signaling. AB - The small protein ubiquitin is a central regulator of a cell's life and death. Ubiquitin is best known for targeting protein destruction by the 26S proteasome. In the past few years, however, nonproteolytic functions of ubiquitin have been uncovered at a rapid pace. These functions include membrane trafficking, protein kinase activation, DNA repair, and chromatin dynamics. A common mechanism underlying these functions is that ubiquitin, or polyubiquitin chains, serves as a signal to recruit proteins harboring ubiquitin-binding domains, thereby bringing together ubiquitinated proteins and ubiquitin receptors to execute specific biological functions. Recent advances in understanding ubiquitination in protein kinase activation and DNA repair are discussed to illustrate the nonproteolytic functions of ubiquitin in cell signaling. PMID- 19217401 TI - Structural basis of murein peptide specificity of a gamma-D-glutamyl-l-diamino acid endopeptidase. AB - The crystal structures of two homologous endopeptidases from cyanobacteria Anabaena variabilis and Nostoc punctiforme were determined at 1.05 and 1.60 A resolution, respectively, and contain a bacterial SH3-like domain (SH3b) and a ubiquitous cell-wall-associated NlpC/P60 (or CHAP) cysteine peptidase domain. The NlpC/P60 domain is a primitive, papain-like peptidase in the CA clan of cysteine peptidases with a Cys126/His176/His188 catalytic triad and a conserved catalytic core. We deduced from structure and sequence analysis, and then experimentally, that these two proteins act as gamma-D-glutamyl-L-diamino acid endopeptidases (EC 3.4.22.-). The active site is located near the interface between the SH3b and NlpC/P60 domains, where the SH3b domain may help define substrate specificity, instead of functioning as a targeting domain, so that only muropeptides with an N terminal L-alanine can bind to the active site. PMID- 19217403 TI - Fission yeast Scm3 mediates stable assembly of Cnp1/CENP-A into centromeric chromatin. AB - Mis16 and Mis18 are subunits of a protein complex required for incorporation of the histone H3 variant CenH3 (Cnp1/CENP-A) into centromeric chromatin in Schizosaccharomyces pombe and mammals. How the Mis16-Mis18 complex performs this function is unknown. Here, we report that the Mis16-Mis18 complex is required for centromere localization of Scm3(Sp), a Cnp1-binding protein related to Saccharomyces cerevisiae Scm3. Scm3(Sp) is required for centromeric localization of Cnp1, while Scm3(Sp) localizes at centromeres independently of Cnp1. Like the Mis16-Mis18 complex but unlike Cnp1, Scm3(Sp) dissociates from centromeres during mitosis. Inactivation of Scm3(Sp) or Mis18 increases centromere localization of histones H3 and H2A/H2B, which are largely absent from centromeres in wild-type cells. Whereas S. cerevisiae Scm3 is proposed to replace histone H2A/H2B in centromeric nucleosomes, the dynamic behavior of S. pombe Scm3 suggests that it acts as a Cnp1 assembly/maintenance factor that directly mediates the stable deposition of Cnp1 into centromeric chromatin. PMID- 19217404 TI - Fission yeast Scm3: A CENP-A receptor required for integrity of subkinetochore chromatin. AB - The mechanisms ensuring specific incorporation of CENP-A at centromeres are poorly understood. Mis16 and Mis18 are required for CENP-A localization at centromeres and form a complex that is conserved from fission yeast to human. Fission yeast sim1 mutants that alleviate kinetochore domain silencing are defective in Scm3(Sp), the ortholog of budding yeast Scm3(Sc). Scm3(Sp) depends on Mis16/18 for its centromere localization and like them is recruited to centromeres in late anaphase. Importantly, Scm3(Sp) coaffinity purifies with CENP A(Cnp1) and associates with CENP-A(Cnp1) in vitro, yet localizes independently of intact CENP-A(Cnp1) chromatin and is differentially released from chromatin. While Scm3(Sc) has been proposed to form a unique hexameric nucleosome with CENP A(Cse4) and histone H4 at budding yeast point centromeres, we favor a model in which Scm3(Sp) acts as a CENP-A(Cnp1) receptor/assembly factor, cooperating with Mis16 and Mis18 to receive CENP-A(Cnp1) from the Sim3 escort and mediate assembly of CENP-A(Cnp1) into subkinetochore chromatin. PMID- 19217406 TI - Reconstitution of yeast silent chromatin: multiple contact sites and O-AADPR binding load SIR complexes onto nucleosomes in vitro. AB - At yeast telomeres and silent mating-type loci, chromatin assumes a higher-order structure that represses transcription by means of the histone deacetylase Sir2 and structural proteins Sir3 and Sir4. Here, we present a fully reconstituted system to analyze SIR holocomplex binding to nucleosomal arrays. Purified Sir2-3 4 heterotrimers bind chromatin, cooperatively yielding a stable complex of homogeneous molecular weight. Remarkably, Sir2-3-4 also binds naked DNA, reflecting the strong, albeit nonspecific, DNA-binding activity of Sir4. The binding of Sir3 to nucleosomes is sensitive to histone H4 N-terminal tail removal, while that of Sir2-4 is not. Dot1-mediated methylation of histone H3K79 reduces the binding of both Sir3 and Sir2-3-4. Additionally, a byproduct of Sir2 mediated NAD hydrolysis, O-acetyl-ADP-ribose, increases the efficiency with which Sir3 and Sir2-3-4 bind nucleosomes. Thus, in small cumulative steps, each Sir protein, unmodified histone domains, and contacts with DNA contribute to the stability of the silent chromatin complex. PMID- 19217405 TI - Rif1 and rif2 inhibit localization of tel1 to DNA ends. AB - Chromosome ends, known as telomeres, have to be distinguished from DNA double strand breaks (DSBs) that activate the DNA-damage checkpoint. In budding yeast, the ATM homolog Tel1 associates preferentially with short telomeres and promotes telomere addition. Here, we show that the telomeric proteins Rif1 and Rif2 attenuate Tel1 recruitment to DNA ends through distinct mechanisms. Both Rif1 and Rif2 inhibit the localization of Tel1, but not the Mre11-Rad50-Xrs2 (MRX) complex, to adjacent DNA ends. Rif1 function is weaker at short telomeric repeats compared with Rif2 function and is partly dependent on Rif2. Rif2 competes with Tel1 for binding to the C terminus of Xrs2. Once Tel1 is delocalized, MRX does not associate efficiently with Rap1-covered DNA ends. These results reveal a mechanism by which telomeric DNA sequences mask DNA ends from Tel1 recognition for the regulation of telomere length. PMID- 19217407 TI - Chromosome-wide Rad51 spreading and SUMO-H2A.Z-dependent chromosome fixation in response to a persistent DNA double-strand break. AB - DNA double-strand breaks (DSBs) are acutely hazardous for cells, as they can cause genome instability. DSB repair involves the sequential recruitment of repair factors to the DSBs, followed by Rad51-mediated homology probing, DNA synthesis, and ligation. However, little is known about how cells react if no homology is found and DSBs persist. Here, by monitoring a single persistent DNA break, we show that, following DNA resection and RPA recruitment, Rad51 spreads chromosome-wide bidirectionally from the DSB but selectively only on the broken chromosome. Remarkably, the persistent DSB is later fixed to the nuclear periphery in a process that requires Rad51, the histone variant H2A.Z, its SUMO modification, and the DNA-damage checkpoint. Indeed, H2A.Z is deposited close to the break early but transiently and directs DNA resection, single DSB-induced checkpoint activation, and DSB anchoring. Thus, a persistent DSB induces a multifaceted response, which is linked to a specific chromatin mark. PMID- 19217408 TI - TAF12 recruits Gadd45a and the nucleotide excision repair complex to the promoter of rRNA genes leading to active DNA demethylation. AB - Many studies have detailed the repressive effects of DNA methylation on gene expression. However, the mechanisms that promote active demethylation are just beginning to emerge. Here, we show that methylation of the rDNA promoter is a dynamic and reversible process. Demethylation of rDNA is initiated by recruitment of Gadd45a (growth arrest and DNA damage inducible protein 45 alpha) to the rDNA promoter by TAF12, a TBP-associated factor that is contained in Pol I- and Pol II specific TBP-TAF complexes. Once targeted to rDNA, Gadd45a triggers demethylation of promoter-proximal DNA by recruiting the nucleotide excision repair (NER) machinery to remove methylated cytosines. Knockdown of Gadd45a, XPA, XPG, XPF, or TAF12 or treatment with drugs that inhibit NER causes hypermethylation of rDNA, establishes heterochromatic histone marks, and impairs transcription. The results reveal a mechanism that recruits the DNA repair machinery to the promoter of active genes, keeping them in a hypomethylated state. PMID- 19217409 TI - Transcriptional termination enhances protein expression in human cells. AB - Transcriptional termination of mammalian RNA polymerase II (Pol II) requires a poly(A) (pA) signal and, often, a downstream terminator sequence. Termination is triggered following recognition of the pA signal by Pol II and subsequent pre mRNA cleavage, which occurs either at the pA site or in transcripts from terminator elements. Although this process has been extensively studied, it is generally considered inconsequential to the level of gene expression. However, our results demonstrate that termination acts as a driving force for optimal gene expression. We show that this effect is general but most dramatic where weak or noncanonical pA signals are present. We establish that termination of Pol II increases the efficiency of pre-mRNA processing that is completed posttranscriptionally. As such, transcripts escape from nuclear surveillance. PMID- 19217410 TI - Molecular architecture of the human pre-mRNA 3' processing complex. AB - Pre-mRNA 3' end formation is an essential step in eukaryotic gene expression. Over half of human genes produce alternatively polyadenylated mRNAs, suggesting that regulated polyadenylation is an important mechanism for posttranscriptional gene control. Although a number of mammalian mRNA 3' processing factors have been identified, the full protein composition of the 3' processing machinery has not been determined, and its structure is unknown. Here we report the purification and subsequent proteomic and structural characterization of human mRNA 3' processing complexes. Remarkably, the purified 3' processing complex contains approximately 85 proteins, including known and new core 3' processing factors and over 50 proteins that may mediate crosstalk with other processes. Electron microscopic analyses show that the core 3' processing complex has a distinct "kidney" shape and is approximately 250 A in length. Together, our data has revealed the complexity and molecular architecture of the pre-mRNA 3' processing complex. PMID- 19217411 TI - Bad targets the permeability transition pore independent of Bax or Bak to switch between Ca2+-dependent cell survival and death. AB - Calcium oscillations exert physiological control on mitochondrial energy metabolism and can also lead to mitochondrial membrane permeabilization and cell death. The outcome of the mitochondrial calcium signaling is altered by stress factors such as ceramide or staurosporine. However, the mechanism of this proapoptotic switch remains unclear. Using genetic, biochemical, pharmacological, and functional approaches, we here show that ceramide and staurosporine target PP2A and protein kinases A and C, respectively, in a mitochondria-associated signaling complex to induce dephosphorylation of the BH3-only protein Bad. Dephosphorylated Bad sensitizes the mitochondrial permeability transition pore (PTP) to Ca2+ through a Bcl-xL-sensitive and VDAC-mediated process. Furthermore, the Bad-induced sensitization of the PTP to Ca2+ does not require Bax or Bak. Thus, phospho-regulatory mechanisms converge on Bad to switch between the survival and apoptotic functions of mitochondrial calcium signaling by activating a mechanism whereby a BH3-only protein bypasses Bax/Bak and engages the PTP. PMID- 19217412 TI - Hsm3/S5b participates in the assembly pathway of the 19S regulatory particle of the proteasome. AB - The 26S proteasome, the central enzyme of the ubiquitin-proteasome system, is comprised of the 20S catalytic core particle (CP) and the 19S regulatory particle (RP), itself composed of two subcomplexes, the base and the lid. 20S proteasome assembly is assisted by several chaperones. Integral subunits of the RP participate in its assembly, but no external factors have been identified so far. Here we characterize the yeast Hsm3 protein, which displays unique features regarding 19S assembly. Hsm3 associates with 19S subcomplexes via a carboxy terminal domain of the Rpt1 base subunit but is missing in the final 26S proteasome. Moreover, Hsm3 is specifically required for the base subcomplex assembly. Finally, we identify the putative species-specific 19S subunit S5b as a functional homolog of the Hsm3 chaperone in mammals. These findings shed light on chaperone-assisted proteasome assembly in eukaryotes. PMID- 19217413 TI - Phospho-regulated SUMO interaction modules connect the SUMO system to CK2 signaling. AB - Attachment of SUMO to proteins regulates protein-protein interactions through noncovalent binding of the SUMO moiety to specialized SUMO interaction motifs (SIMs). A core of hydrophobic amino acids has been described as the major determinant of SIM function. Using the transcriptional coregulator and SUMO ligase PIAS1 as a model, we define an extended phospho-regulated SIM module. We show that serine residues adjacent to the hydrophobic core are phosphorylated by CK2 and demonstrate that this dictates binding of free SUMO and SUMO conjugates to PIAS1 in vivo. We provide evidence that the phosphorylated residues contact lysine 39 and 35 in SUMO1 and SUMO2, respectively. Phospho-dependent SUMO binding does not impair the ligase activity but affects the transcriptional coregulatory potential of PIAS1 and other PIAS family members. CK2-regulated phosphoSIM modules were also dissected in the tumor suppressor PML and the exosome component PMSCL1, indicating that these modules serve as general platforms that integrate CK2- and SUMO-regulated signaling networks. PMID- 19217414 TI - Reviews in developmental cell: vascular biology and more. PMID- 19217415 TI - A top-NOTCH way to make astrocytes. AB - During mammalian nervous system development, neural precursor cells first generate neurons and then switch to astrocytes. In this issue of Developmental Cell, Namihira and colleagues show that committed neuronal precursors and young neurons promote the production of astrocytes from the remaining neural precursors by activating Notch signaling, leading to demethylation and upregulation of astrocyte-specific genes via cytokine-mediated JAK-STAT signaling. PMID- 19217416 TI - Calcium flickers lighting the way in chemotaxis? AB - Recent studies identified local calcium signals at the leading edge of migrating cells, suggesting a new role for calcium in cell polarization and chemotaxis. PMID- 19217418 TI - A new G(q)-initiated MAPK signaling pathway in the heart. AB - Pathological cardiac hypertrophy involves auto/paracrine mediators acting through G(q/11)-coupled receptors. A novel signaling route stimulated by betagamma subunits of G(q/11) results in the autophosphorylation of ERK1/2 on a new site and the nuclear retention of ERK1/2, thereby activating hypertrophic gene programs. PMID- 19217417 TI - Fibroblasts inform the heart: control of cardiomyocyte cycling and size by age dependent paracrine signals. AB - Within the developing and adult heart, the fibroblast is often dismissed as merely a structural element, important just to mechanical integrity or to scarring when excessive in disease. Ieda et al. in this issue of Developmental Cell now report an essential program of paracrine factor production in cardiac fibroblasts that controls heart muscle cell growth, driving cycling or enlargement depending on the fibroblasts' developmental stage. PMID- 19217419 TI - SUMO weighs in on a photoreceptor finish. AB - In the developing retina the nuclear hormone receptor Nr2e3 has dual roles, promoting expression of rod-specific genes and repressing expression of cone specific genes. In a recent issue of Neuron, Onishi et al. show that PIAS3 mediated SUMOylation of Nr2e3 plays an essential role in the specification of rod photoreceptors by converting Nr2e3 to a potent repressor of cone gene expression. PMID- 19217420 TI - Regulation of angiogenesis by oxygen and metabolism. AB - Blood vessels form an important interface between the environment and the organism by carrying oxygen and nutrients to all cells and thus determining cellular metabolism. It is therefore not surprising that oxygen and metabolism influence the development of the vascular network. Here, we discuss recent insights regarding the emerging crosstalk between angiogenesis and metabolism. We will highlight advances in how oxygen and metabolism regulate angiogenesis as well as how angiogenic factors in turn also regulate metabolism. PMID- 19217422 TI - Angiogenesis: a team effort coordinated by notch. AB - The past two decades of angiogenesis research have identified a wealth of pro- and antiangiogenic signals originating from the tissue environment, which control blood vessel density and function. Understanding when and how blood vessels respond to the combination of signals they encounter to achieve a balanced cellular response is a major challenge for the field of developmental and tumor angiogenesis. This review focuses on how endothelial cell-cell communication via the Notch pathway contributes to this signal integration and is essential for functional vessel patterning. PMID- 19217421 TI - Transcriptional control of endothelial cell development. AB - The transcription factors that regulate endothelial cell development have been a focus of active research for several years, and many players in the endothelial transcriptional program have been identified. This review discusses the function of several major regulators of endothelial transcription, including members of the Sox, Ets, Forkhead, GATA, and Kruppel-like families. This review also highlights recent developments aimed at unraveling the combinatorial mechanisms and transcription factor interactions that regulate endothelial cell specification and differentiation during vasculogenesis and angiogenesis. PMID- 19217423 TI - The control of vascular integrity by endothelial cell junctions: molecular basis and pathological implications. AB - Human pathologies such as vascular malformations, hemorrhagic stroke, and edema have been associated with defects in the organization of endothelial cell junctions. Understanding the molecular basis of these diseases requires different integrated approaches which include basic cell biology, clinical studies, and studies in animal models such as mice and zebrafish. In this review we discuss recent findings derived from these approaches and their possible integration in a common picture. PMID- 19217424 TI - Cellular and molecular mechanisms of vascular lumen formation. AB - The formation of vascular lumens by endothelial cells is a critical step in the angiogenic process that occurs during invasion and growth of the incipient vascular sprout. Once a lumen is established, capillaries are rapidly exposed to the physical forces associated with the flow of blood which, together with genetic information, regulate the ultimate size of inner vessel diameter. Here we review the recent literature on vascular lumen formation and compare it to lumen formation in other epithelial systems. We also discuss the regulation of lumen diameter after vascular morphogenesis has been completed. PMID- 19217426 TI - Committed neuronal precursors confer astrocytic potential on residual neural precursor cells. AB - During midgestation, mammalian neural precursor cells (NPCs) differentiate only into neurons. Generation of astrocytes is prevented at this stage, because astrocyte-specific gene promoters are methylated. How the subsequent switch from suppression to expression of astrocytic genes occurs is unknown. We show in this study that Notch ligands are expressed on committed neuronal precursors and young neurons in mid-gestational telencephalon, and that neighboring Notch-activated NPCs acquire the potential to become astrocytes. Activation of the Notch signaling pathway in midgestational NPCs induces expression of the transcription factor nuclear factor I, which binds to astrocytic gene promoters, resulting in demethylation of astrocyte-specific genes. These findings provide a mechanistic explanation for why neurons come first: committed neuronal precursors and young neurons potentiate remaining NPCs to differentiate into the next cell lineage, astrocytes. PMID- 19217425 TI - Cardiac fibroblasts regulate myocardial proliferation through beta1 integrin signaling. AB - Growth and expansion of ventricular chambers is essential during heart development and is achieved by proliferation of cardiac progenitors. Adult cardiomyocytes, by contrast, achieve growth through hypertrophy rather than hyperplasia. Although epicardial-derived signals may contribute to the proliferative process in myocytes, the factors and cell types responsible for development of the ventricular myocardial thickness are unclear. Using a coculture system, we found that embryonic cardiac fibroblasts induced proliferation of cardiomyocytes, in contrast to adult cardiac fibroblasts that promoted myocyte hypertrophy. We identified fibronectin, collagen, and heparin binding EGF-like growth factor as embryonic cardiac fibroblast-specific signals that collaboratively promoted cardiomyocyte proliferation in a paracrine fashion. Myocardial beta1-integrin was required for this proliferative response, and ventricular cardiomyocyte-specific deletion of beta1-integrin in mice resulted in reduced myocardial proliferation and impaired ventricular compaction. These findings reveal a previously unrecognized paracrine function of embryonic cardiac fibroblasts in regulating cardiomyocyte proliferation. PMID- 19217429 TI - A systematic analysis of Tinman function reveals Eya and JAK-STAT signaling as essential regulators of muscle development. AB - Nk-2 proteins are essential developmental regulators from flies to humans. In Drosophila, the family member tinman is the major regulator of cell fate within the dorsal mesoderm, including heart, visceral, and dorsal somatic muscle. To decipher Tinman's direct regulatory role, we performed a time course of ChIP-on chip experiments, revealing a more prominent role in somatic muscle specification than previously anticipated. Through the combination of transgenic enhancer reporter assays, colocalization studies, and phenotypic analyses, we uncovered two additional factors within this myogenic network: by activating eyes absent, Tinman's regulatory network extends beyond developmental stages and tissues where it is expressed; by regulating stat92E expression, Tinman modulates the transcriptional readout of JAK/STAT signaling. We show that this pathway is essential for somatic muscle development in Drosophila and for myotome morphogenesis in zebrafish. Taken together, these data uncover a conserved requirement for JAK/STAT signaling and an important component of the transcriptional network driving myogenesis. PMID- 19217427 TI - AMP-activated protein kinase phosphorylates retinoblastoma protein to control mammalian brain development. AB - AMP-activated protein kinase (AMPK) is an evolutionarily conserved metabolic sensor that responds to alterations in cellular energy levels to maintain energy balance. While its role in metabolic homeostasis is well documented, its role in mammalian development is less clear. Here we demonstrate that mutant mice lacking the regulatory AMPK beta1 subunit have profound brain abnormalities. The beta1(-/ ) mice show atrophy of the dentate gyrus and cerebellum, and severe loss of neurons, oligodendrocytes, and myelination throughout the central nervous system. These abnormalities stem from reduced AMPK activity, with ensuing cell cycle defects in neural stem and progenitor cells (NPCs). The beta1(-/-) NPC deficits result from hypophosphorylation of the retinoblastoma protein (Rb), which is directly phosphorylated by AMPK at Ser(804). The AMPK-Rb axis is utilized by both growth factors and energy restriction to increase NPC growth. Our results reveal that AMPK integrates growth factor signaling with cell cycle control to regulate brain development. PMID- 19217428 TI - Interactions with the Abelson tyrosine kinase reveal compartmentalization of eyes absent function between nucleus and cytoplasm. AB - Eyes absent (Eya), named for its role in Drosophila eye development but broadly conserved in metazoa, possesses dual functions as a transcriptional coactivator and protein tyrosine phosphatase. Although Eya's transcriptional activity has been extensively characterized, the physiological requirements for its phosphatase activity remain obscure. In this study, we provide insight into Eya's participation in phosphotyrosine-mediated signaling networks by demonstrating cooperative interactions between Eya and the Abelson (Abl) tyrosine kinase during development of the Drosophila larval visual system. Mechanistically, Abl-mediated phosphorylation recruits Eya to the cytoplasm, where in vivo studies reveal a requirement for its phosphatase function. Thus, we propose a model in which, in addition to its role as a transcription factor, Eya functions as a cytoplasmic protein tyrosine phosphatase. PMID- 19217430 TI - Displacement of formins from growing barbed ends by bud14 is critical for actin cable architecture and function. AB - Normal cellular development and function require tight spatiotemporal control of actin assembly. Formins are potent actin assembly factors that protect the growing ends of actin filaments from capping proteins. However, it is unresolved how the duration of formin-mediated actin assembly events is controlled, whether formins are actively displaced from growing ends, and how filament length is regulated in vivo. Here, we identify Bud14 as a high-affinity inhibitor of the yeast formin Bnr1 that rapidly displaces the Bnr1 FH2 domain from growing barbed ends. Consistent with these activities, bud14Delta cells display fewer actin cables, which are aberrantly long, bent, and latrunculinA resistant, leading to defects in secretory vesicle movement. Moreover, bud14Delta suppressed mutations that cause abnormally numerous and shortened cables, restoring wild-type actin architecture. From these results, we propose that formin displacement factors regulate filament length and are required in vivo to maintain proper actin network architecture and function. PMID- 19217431 TI - Increased PDGFRalpha activation disrupts connective tissue development and drives systemic fibrosis. AB - PDGF signaling regulates the development of mesenchymal cell types in the embryo and in the adult, but the role of receptor activation in tissue homeostasis has not been investigated. We have generated conditional knockin mice with mutations in PDGFRalpha that drive increased kinase activity under the control of the endogenous PDGFRalpha promoter. In embryos, increased PDGFRalpha signaling leads to hyperplasia of stromal fibroblasts, which disturbs normal smooth muscle tissue in radially patterned organs. In adult mice, elevated PDGFRalpha signaling also increases connective tissue growth, leading to a progressive fibrosis phenotype in multiple organs. Increased PDGFRalpha signaling in an Ink4a/Arf-deficient genetic background leads to accelerated fibrosis, suggesting a new role for tumor suppressors in attenuating fibrotic diseases. These results highlight the role of PDGFRalpha in normal connective tissue development and homeostasis and demonstrate a pivotal role for PDGFRalpha signaling in systemic fibrosis diseases. PMID- 19217432 TI - Inactivation of murine Usp1 results in genomic instability and a Fanconi anemia phenotype. AB - Fanconi anemia (FA) is a human genetic disease characterized by chromosome instability, cancer predisposition, and cellular hypersensitivity to DNA crosslinking agents. The FA pathway regulates the repair of DNA crosslinks. A critical step in this pathway is the monoubiquitination and deubiquitination of FANCD2. Deubiquitination of FANCD2 is mediated by the ubiquitin protease, USP1. Here, we demonstrate that targeted deletion of mouse Usp1 results in elevated perinatal lethality, male infertility, crosslinker hypersensitivity, and an FA phenotype. Usp1(-/-) mouse embryonic fibroblasts had heightened levels of monoubiquitinated Fancd2 in chromatin. Usp1(-/-) cells exhibited impaired Fancd2 foci assembly and a defect in homologous recombination repair. Double knockout of Usp1 and Fancd2 resulted in a more severe phenotype than either single knockout. Our results indicate that mouse Usp1 functions downstream in the FA pathway. Deubiquitination is a critical event required for Fancd2 nuclear foci assembly, release from chromatin, and function in DNA repair. PMID- 19217434 TI - Protracted elimination of gold nanoparticles from mouse liver. AB - The present study aims at revealing the fate of 40-nm gold nanoparticles after intravenous injections. The gold nanoparticles were traced histochemically with light and transmission electron microscopy using autometallographic (AMG) staining, and the gold content in the liver was determined with inductively coupled plasma mass spectrometry (ICP-MS). Gold nanoparticles were identified in almost all Kupffer cells one day after the injection, but the fraction of gold loaded cells gradually decreased to about one fifth after 6 months. Transmission electron microscopic analysis showed that the gold nanoparticles had accumulated inside the vesicular lysosome/endosome-like structures of the macrophages. At day 1, about 4.5 per thousand of the area of the liver sections was AMG-stained, after 1 month it had decreased to 0.7 per thousand, and thereafter no further significant reduction was recorded. Because ICP-MS only showed a 9% fall in the gold content over the observed 6 months, the AMG finding of a significant reduction in the stained area of the liver sections and number of macrophages loaded with gold nanoparticles reveals that over time an increasing part of the total amount of gold nanoparticles in the liver is contained in fewer macrophages accumulated in growing clusters. PMID- 19217435 TI - Effectiveness of school-based prevention and intervention programs for children and adolescents with emotional disturbance: a meta-analysis. AB - The present study evaluated the effectiveness of school-based prevention and intervention programs for children and adolescents at-risk for and with emotional disturbance. Published outcome studies (k=29) from December, 1988, to March, 2006, including 1405 children and adolescents were reviewed. Each investigation was coded on several variables describing the child, parent, and teacher samples, as well as reported outcome results. The overall mean weighted effect size was 1.00 at post-test and 1.35 at follow-up. Mean weighted ESs were 0.42 for between subjects design studies, 0.87 for within-subjects design studies, and 1.87 for single-subject design studies. Prevention programs yielded a mean weighted ES of 0.54 and intervention programs produced a mean weighted ES of 1.35. Findings for specific outcome foci are presented and implications are discussed. PMID- 19217433 TI - Bdnf overexpression in hippocampal neurons prevents dendritic atrophy caused by Rett-associated MECP2 mutations. AB - The expression of the methylated DNA-binding protein MeCP2 increases during neuronal development, which suggests that this epigenetic factor is crucial for neuronal terminal differentiation. We evaluated dendritic and axonal development in embryonic day-18 hippocampal neurons in culture by measuring total length and counting branch point numbers at 4 days in vitro, well before synapse formation. Pyramidal neurons transfected with a plasmid encoding a small hairpin RNA (shRNA) to knockdown endogenous Mecp2 had shorter dendrites than control untransfected neurons, without detectable changes in axonal morphology. On the other hand, overexpression of wildtype (wt) human MECP2 increased dendritic branching, in addition to axonal branching and length. Consistent with reduced neuronal growth and complexity in Rett syndrome (RTT) brains, overexpression of human MECP2 carrying missense mutations common in RTT individuals (R106W or T158M) reduced dendritic and axonal length. One of the targets of MeCP2 transcriptional control is the Bdnf gene. Indeed, endogenous Mecp2 knockdown increased the intracellular levels of BDNF protein compared to untransfected neurons, suggesting that MeCP2 represses Bdnf transcription. Surprisingly, overexpression of wt MECP2 also increased BDNF levels, while overexpression of RTT-associated MECP2 mutants failed to affect BDNF levels. The extracellular BDNF scavenger TrkB-Fc prevented dendritic overgrowth in wt MECP2-overexpressing neurons, while overexpression of the Bdnf gene reverted the dendritic atrophy caused by Mecp2-knockdown. However, this effect was only partial, since Bdnf increased dendritic length only to control levels in mutant MECP2-overexpressing neurons, but not as much as in Bdnf transfected cells. Our results demonstrate that MeCP2 plays varied roles in dendritic and axonal development during neuronal terminal differentiation, and that some of these effects are mediated by autocrine actions of BDNF. PMID- 19217436 TI - Collective motivation beliefs of early adolescents working in small groups. AB - This study examined collective efficacy, group cohesion, and group performance in 125 randomly assigned groups of older (mean age 13.45 years) and younger (mean age 11.41 years) early adolescents working on three cooperative tasks. Collective motivation significantly predicted performance, even after controlling for past performance and self-efficacy for the older but not the younger participants. For the older (but not the younger) participants, groups with high collective efficacy and group cohesion scored higher on performance tasks than groups with low collective efficacy and group cohesion. The results point to the emergence of collective motivation beliefs in early adolescence, consistent with theories of social and cognitive development. Implications for theory and suggestions for future research are discussed. PMID- 19217437 TI - A preliminary investigation of within-year growth patterns in reading and mathematics curriculum-based measures. AB - This study examined patterns of growth across benchmark assessments for curriculum-based measures (CBM) over 2 academic years, with the twofold purpose of replicating earlier findings of growth patterns on R-CBM and conducting a preliminary investigation of growth patterns on M-CBM and CBM-Maze. The sample included 898 cases from 3rd through 5th grade over 2 academic years from one elementary school in the rural Midwest. All students participated in tri-annual benchmark assessments in which they were administered R-CBM, CBM-Maze, and M-CBM. Repeated measures analysis of variance was used to examine patterns of growth for all three measures. Students demonstrated greater R-CBM weekly growth during winter-to-spring than fall-to-winter across grade levels. Patterns for CBM-Maze and M-CBM were somewhat inconsistent across school years and grade levels. Results are discussed in terms of contextual variables that may impact within year growth patterns and technical characteristics of CBM slopes. PMID- 19217438 TI - Preliminary report: Serum levels of retinol-binding protein 4 in preeclampsia. AB - The objective of the study was to investigate serum levels of the adipokine retinol-binding protein 4 (RBP4) in patients with preeclampsia (PE) as compared with healthy controls of similar gestational age. Retinol-binding protein 4 serum levels were quantified by enzyme-linked immunosorbent assay in control (n = 20) and PE (n = 16) patients. Mean maternal RBP4 concentrations were not significantly different in PE (24.5 mg/L) as compared with controls (22.3 mg/L). Furthermore, RBP4 did not correlate to clinical and biochemical measures of pregnancy outcome, renal function, glucose, and lipid metabolism, as well as inflammation. Our results do not support a role of RBP4 in the pathogenesis of PE. PMID- 19217439 TI - Evaluation of endogenous nitric oxide synthesis in congenital urea cycle enzyme defects. AB - Nitric oxide (NO) is synthesized from arginine and O(2) by nitric oxide synthase (NOS). Citrulline, which is formed as a by-product of the NOS reaction, can be recycled to arginine by the 2 enzymes acting in the urea cycle: argininosuccinate synthetase (ASS) and argininosuccinate lyase (ASL). Although the complete urea cycle is expressed only in the liver, ASS and ASL are expressed in other organs including the kidney and vascular endothelium. To examine possible alterations of the NO pathway in urea cycle defects, we measured plasma concentrations of arginine and citrulline and serum concentrations of nitrite/nitrate (NOx(-), stable NO metabolites) and asymmetric dimethylarginine (ADMA, an endogenous NOS inhibitor) in patients with congenital urea cycle disorders of 3 types: ornithine transcarbamylase (OTC) deficiency, ASS deficiency, and ASL deficiency. All were receiving oral arginine replacement at the time of this study. The same parameters were also measured in healthy subjects, who participated as controls. The OTC-deficient patients had significantly high NOx(-) and nonsignificantly high ADMA concentrations. Their NOx(-) was significantly positively correlated with arginine. The ASS-deficient patients had significantly low NOx(-) and significantly high ADMA concentrations. The ASL-deficient patients had normal NOx(-) and nonsignificantly high ADMA concentrations. In ASS-deficient and ASL deficient patients, the NOx(-) was significantly inversely correlated with citrulline. These results suggest that NO synthesis is enhanced in OTC-deficient patients while receiving arginine but that NO synthesis remains low in ASS deficient patients despite receiving arginine. They also suggest that endogenous NO synthesis is negatively affected by citrulline and ADMA in ASS-deficient and ASL-deficient patients. Although the molecular mechanisms remain poorly understood, we infer that the NO pathway might play a role in the pathophysiology related to congenital urea cycle disorders. PMID- 19217440 TI - Influence of cholesteryl ester transfer protein, peroxisome proliferator activated receptor alpha, apolipoprotein E, and apolipoprotein A-I polymorphisms on high-density lipoprotein cholesterol, apolipoprotein A-I, lipoprotein A-I, and lipoprotein A-I:A-II concentrations: the Prospective Epidemiological Study of Myocardial Infarction study. AB - The plasma level of high-density lipoprotein cholesterol (HDL-C) is known to be inversely associated with cardiovascular risk. However, besides lifestyle, gene polymorphism may influence the HDL-C concentration. The aim of this study was to investigate the possibility of interactions between CETP, PPARA, APOE, and APOAI polymorphisms and HDL-C, apolipoprotein (apo) A-I, lipoprotein (Lp) A-I, and Lp A I:A-II in a sample selected from the Prospective Epidemiological Study of Myocardial Infarction (PRIME) study population who remained free of cardiovascular events over 5 years of follow-up. Healthy individuals (857) were randomly selected for genotyping the PRIME study subjects. The population was selected so as to provide 25% of subjects in the lowest tertile of HDL-C (< or = 28 mg/dL) in the whole PRIME study sample, 25% of subjects in the highest tertile of HDL-C (> or = 73 mg/dL), and 50% of subjects in the medium tertile of HDL-C (28-73 mg/dL). Genotyping was performed by using a polymerase chain reaction system with predeveloped TaqMan allelic discrimination assay. The CETP A373P rare allele c was less frequent in the group of subjects with high HDL-C, apo A-I, Lp A-I, and Lp A-I:A-II concentrations. Apolipoprotein A-I and Lp A-I were also found to be higher in the presence of the epsilon2 allele coding for APOE. The effect of the CETP A373P rare allele c on HDL-C was independent of all tested parameters except triglycerides. The respective effect of these polymorphisms and triglycerides on cardiovascular risk should be evaluated prospectively. PMID- 19217441 TI - Homeostasis model assessment of insulin resistance*body mass index interactions at ages 9 to 10 years predict metabolic syndrome risk factor aggregate score at ages 18 to 19 years: a 10-year prospective study of black and white girls. AB - If homeostasis model assessment of insulin resistance (HOMA-IR) interactions with obesity (body mass index [BMI]) at ages 9 to 10 years predict aggregate metabolic syndrome risk factors at ages 18 to 19 years, this would identify novel avenues for primary prevention of metabolic syndrome. Our hypothesis was that HOMA-IR*BMI interactions at ages 9 to 10 years would predict aggregate metabolic syndrome risk factor z scores at ages 18 to 19 years in prospective studies of a biracial population of girls. Two centers in the National Heart, Lung, and Blood Institute Growth and Health Study measured serum insulin and glucose at ages 9 to 10 years and 5 metabolic syndrome risk factors at ages 18 to 19 years (triglyceride, high density lipoprotein cholesterol, systolic/diastolic blood pressure, waist circumference, and glucose). Studies in Cincinnati, OH, included girls from public and parochial schools in the inner city, within-city residential neighborhoods, and suburban areas; and those in Washington, DC, included girls from a health maintenance organization. Girls (194 white, 281 black) were studied first at ages 9 to 10 years, then at ages 18 to 19 years. We assessed HOMA-IR*BMI interactions at ages 9 to 10 years with race-specific z scores for 5 metabolic syndrome risk factors at ages 18 to 19 years. The lowest summed z score (mean +/- SD) was observed for subjects in the lowest tertiles for both HOMA-IR and BMI ( 1.15 +/- 2.05), and the highest z score (2.58 +/- 3.11) was for subjects in the highest tertiles for both HOMA-IR and BMI (P < .0001). For the top BMI tertile, there was a progressive increase in z score (increasing risk of metabolic syndrome) as HOMA-IR increased. Interaction of BMI with HOMA-IR at ages 9 to 10 years predicts aggregate metabolic risk score at ages 18 to 19 years, with progressive risk increments within the top BMI tertile as HOMA-IR increases, opening avenues for intervention to reduce both BMI and HOMA-IR at ages 9 to 10 years as a primary approach to prevention of metabolic syndrome at ages 18 to 19 years. PMID- 19217442 TI - A novel model of type 2 diabetes mellitus based on obesity induced by high-fat diet in BDF1 mice. AB - For experimental research on type 2 diabetes mellitus, a diet-induced obesity dependent diabetes model developed using genetically normal animals is essential. However, attempts at feeding a high-fat diet (HFD) to major inbred strains of mice have not resulted in the establishment of an ideal model. Here, we show that BDF1 mice, the F(1) hybrids of C57BL/6 and DBA/2 normal strains, develop HFD induced obesity-dependent diabetes. BDF1 mice fed a HFD gained weight rapidly and developed severe diabetes characterized by hyperglycemia, glucosuria, and elevation of hemoglobin A(1C) levels in 3 to 4 months. The glucose tolerance of the diabetic mice was significantly impaired, and the elevation of plasma insulin after a glucose load was significantly reduced. Isolated pancreatic islets of HFD fed BDF1 mice showed decreased insulin content and a reduced insulin secretory response to higher concentrations of glucose. Immunohistochemical analysis of the pancreas showed reduced staining intensity to insulin and aberrant distribution of glucagon-positive cells in diabetic BDF1 mice. These observations suggest the cause of the diabetes in HFD-fed BDF1 mice to be dysfunction of the pancreatic beta-cells, which do not produce or secrete enough insulin to compensate for insulin resistance. BDF1 mice fed a HFD showing obesity-dependent diabetes are suggested to be an appropriate animal model of type 2 diabetes mellitus. This model would be useful for exploring the mechanism of obesity-dependent type 2 diabetes mellitus and evaluating antiobesity and antidiabetic drugs. PMID- 19217443 TI - Repeatability and reproducibility of the hyperinsulinemic-euglycemic clamp and the tracer dilution technique in a controlled inpatient setting. AB - The objective of the study was to evaluate the reproducibility and repeatability of the combined use of the hyperinsulinemic-euglycemic (H-E) clamp and tracer dilution techniques. Ten nondiabetic men underwent a low-dose (40 mU/[m(2) min]) H-E clamp that was repeated within 3 to 4 days using porcine or human insulin in a double-blinded, randomized, crossover design. Coefficients of variation (CVs) for intraindividual differences and repeatability coefficient were calculated to evaluate reproducibility and repeatability. The Bland and Altman method was used to quantify repeatability. The CVs for intraindividual differences were 5.7% +/- 3.5% for steady-state (SS) insulin; 6.7% +/- 6.2% and 54.2 +/- 38.3% for basal and SS endogenous glucose product (EGP), respectively; and 10.3% +/- 8.5% for total insulin-stimulated glucose disposal (M) values. Basal EGP, SS EGP, and SS glucose and insulin concentrations were similar for the 2 clamps; but glucose infusion rate (P = .02) and M (borderline significant, P = .06) were higher in the first clamp than the second clamp. No significant correlations between mean of differences and average of basal and SS EGP, SS insulin concentration, and M between the 2 clamps were observed. We also found that the different values were less than the repeatability coefficients of these parameters and that the 95% limits of agreement and the interval of repeatability coefficient of these parameters were similar. There were no differences in metabolic responses between clamps when compared by the type of insulin (porcine vs human) infused. Our findings indicate that, although SS EGP has a high CV, the clamp, which measures insulin action (ie, SS insulin, M), and the tracer dilution technique for assessing basal EGP are repeatable and reproducible. Decreased glucose infusion rate and M over a short period in the second clamp may reflect an accumulative effect of continued physical inactivity. PMID- 19217444 TI - Longitudinal changes in abdominal fat distribution with menopause. AB - Increases in abdominal fat have been reported with menopause, but the impact of menopause on abdominal fat distribution (visceral vs subcutaneous) is still unclear. The objective of the study was to determine if abdominal fat content (volume) or distribution is altered with menopause. Magnetic resonance imaging was used to quantify total abdominal, subcutaneous, and visceral fat in 8 healthy women, both in the premenopausal state and 8 years later in the postmenopausal state. Physical activity (PA) and blood lipids were also measured. Body weight and waist circumference did not change with menopause (pre- vs postmenopause: body weight, 63.2 +/- 3.1 vs 63.9 +/- 2.5 kg; waist circumference, 92.1 +/- 4.6 vs 93.4 +/- 3.7 cm); however, total abdominal fat, subcutaneous fat, and visceral fat all significantly (P < .05) increased with menopause (pre- vs postmenopause: total, 27 154 +/- 4268 vs 34 717 +/- 3272 cm(3); subcutaneous, 19 981 +/- 3203 vs 24 918 +/- 2521 cm(3); visceral, 7173 +/- 1611 vs 9798 +/- 1644 cm(3)). Although absolute adiposity changed with menopause, relative fat distribution was not significantly different after menopause (pre- vs postmenopause: subcutaneous, 73% +/- 3% vs 71% +/- 3%; visceral, 26% +/- 3% vs 28% +/- 3%). Lean mass, fat mass, and PA, along with total cholesterol and triglyceride levels, did not change with menopause. High-density lipoprotein and low-density lipoprotein both increased (P < .05), and the ratio of total cholesterol to high-density lipoprotein decreased (P < .05) with menopause. As measured longitudinally with magnetic resonance imaging, total abdominal fat content increased with menopause despite no change in PA, body weight, or waist circumference; however, menopause did not affect the relative abdominal fat distribution in these women. PMID- 19217445 TI - Plasma asymmetric dimethylarginine is related to anticitrullinated protein antibodies in rheumatoid arthritis of short duration. AB - We have recently demonstrated elevated plasma levels of an endogenous nitric oxide synthase inhibitor, asymmetric dimethyl-L-arginine (ADMA), and its association with carotid atherosclerosis in rheumatoid arthritis (RA). Both an elevated risk of myocardial infarction and increased levels of anticitrullinated protein antibodies (ACPAs), specific for RA, had been shown to precede the onset of clinical RA symptoms. Therefore, our aim was to verify the hypothesis that ADMA accumulation might accompany raised ACPAs titers in RA of short duration (< or = 3 years). Twenty patients (16 women, 4 men; mean age, 45 +/- 12 years; mean disease duration, 2.3 +/- 0.5 years) with active RA despite chronic disease modifying antirheumatic medication, free of cardiovascular disease or atherosclerotic risk factors, were studied. Plasma levels of ADMA and its stereoisomer, symmetric dimethyl-L-arginine (SDMA), were assayed by liquid chromatography/tandem mass spectrometry. The ACPAs were measured by a second generation enzyme-linked immunosorbent assay. In addition to routine biochemical assays, plasma concentrations of tumor necrosis factor alpha, monocyte chemoattractant protein-1, and vascular cell adhesion molecule-1 soluble form were analyzed with respective enzyme-linked immunosorbent assays. A significant positive correlation between levels of ACPAs and ADMA (r = .60, P = .005), but not SDMA (r = -.02, P = .9), was found. Neither ADMA nor SDMA was correlated to any of the clinical or biochemical parameters reflecting disease activity and inflammatory activation. Thus, excessive ADMA accumulation accompanies elevated ACPAs levels in patients with RA of short duration free of cardiovascular disease or risk factors. PMID- 19217447 TI - Meal-induced enhancement in insulin sensitivity is not triggered by hyperinsulinemia in rats. AB - Several reports confirmed the phenomenon of postprandial increase in whole-body insulin sensitivity. Although the initial step of this process is unknown, the pivotal role of postprandial hyperinsulinemia has strongly been suggested. The aim of the present study was to determine whether hyperinsulinemia per se induces insulin sensitization in healthy male Wistar rats. Rapid insulin sensitivity test (RIST) were performed in fasted, anesthetized rats before and during stable hyperinsulinemia achieved by hyperinsulinemic euglycemic glucose clamping (HEGC) with insulin infused either through the jugular vein (systemic HEGC) or into the portal circulation (portal HEGC) at a rate of 3 mU/(kg min). Insulin sensitivity expressed by the rapid insulin sensitivity (RIST) index (in milligrams per kilogram) was characterized by the total amount of glucose needed to maintain prestudy blood glucose level succeeding an intravenous bolus infusion of 50 mU/kg insulin over 5 minutes. In fasted animals, the RIST index was 37.4 +/- 3.1 mg/kg. When hyperinsulinemia mimicking the postprandial state was achieved by systemic HEGC, the RIST index (39.7 +/- 10.6 mg/kg) showed no significant changes as compared with the pre-HEGC values. Hyperinsulinemia achieved by portal insulin infusion also failed to modify the RIST index (35.7 +/- 4.3 mg/kg). The results demonstrate that acute hyperinsulinemia, no matter how induced, does not yield any sensitization to the hypoglycemic effect of insulin. PMID- 19217446 TI - Nonesterified fatty acid exposure activates protective and mitogenic pathways in vascular smooth muscle cells by alternate signaling pathways. AB - Vascular smooth muscle cells (VSMC) are dynamic cells exposed to fluctuating concentrations of nutrients on a daily basis. Nonesterified fatty acids (NEFA) have been indicted as potential mediators of atherosclerosis and exaggerated VSMC remodeling observed in diabetes, and in vitro data support a model of VSMC activation by NEFA. However, recent observations suggest that metabolic stressors such as oxidants and NEFA may also simultaneously induce cytoprotective events as part of a homeostatic "off switch." Our group has established that the transcription factor cyclic adenosine monophosphate response element binding protein (CREB) is important for maintenance of VSMC quiescence, differentiation, and survival. We therefore examined whether acute physiologic NEFA exposure would regulate CREB in primary cultures of bovine aortic VSMC and explored the relationship between signaling to the cytoprotective CREB and the activating mitogen-activated protein kinase pathways. In vitro exposure of VSMC to 3 classes of unsaturated NEFA leads to significant acute, transient, dose-dependent, and repeatedly inducible CREB activation. As expected, extracellular signal-regulated kinase, P38 mitogen-activated protein kinase, Akt, Jun N-terminal kinase, and protein kinase C (PKC) pathways are also activated by NEFA. Using a battery of pharmacologic inhibitors and antioxidants, we demonstrate that CREB activation is mediated by a novel PKC isoform and is reactive oxygen species independent, whereas extracellular signal-regulated kinase activation, in contrast, is mediated by reactive oxygen species and is PKC independent. These data suggest parallel and mechanistically distinct stimulation of separate stabilizing and activating pathways in VSMC response to acute NEFA-mediated stress. Furthermore, the down-regulation of CREB in models of chronic metabolic stress reported in the literature would be expected to disrupt this homeostasis and shift the balance toward VSMC activation, consistent with emerging models of atherosclerosis. PMID- 19217448 TI - Acute administration of GPR40 receptor agonist potentiates glucose-stimulated insulin secretion in vivo in the rat. AB - Recently, several in vitro studies have shown that GPR40 receptor activation by free fatty acids (FFAs) results in glucose-dependent insulin secretion. However, whether GPR40 receptor activation results in glucose-dependent insulin secretion in vivo in rats is not known. Therefore, we evaluated the effect of synthetic GPR40 receptor agonist (compound 1) on glucose tolerance test (GTT) in fed, fasted, and insulin-resistant rats. In oral GTT, intraperitoneal GTT, and intravenous GTT, GPR40 receptor agonist improved glucose tolerance, which was associated with increase in plasma insulin level. Interestingly, in GTTs, the rise in insulin levels in agonist-treated group was directly proportional to the rate of rise and peak levels of glucose in control group. Although glibenclamide, a widely used insulin secretagogue, improved glucose tolerance in all GTTs, it did not display insulin release in intraperitoneal GTT or intravenous GTT. In the absence of glucose load, GPR40 receptor agonist did not significantly change the plasma insulin concentration, but did decrease the plasma glucose concentration. Fasted rats exhibited impaired glucose-stimulated insulin secretion (GSIS) as compared with fed rats. Compound 1 potentiated GSIS in fasted state but failed to do so in fed state. Suspecting differential pharmacokinetics, a detailed pharmacokinetic evaluation was performed, which revealed the low plasma concentration of compound 1 in fed state. Consequently, we examined the absorption profile of compound 1 at higher doses in fed state; and at a dose at which its absorption was comparable with that in fasted state, we observed significant potentiation of GSIS. Chronic high-fructose (60%) diet feeding resulted in impaired glucose tolerance, which was improved by GPR40 receptor agonist. Therefore, our results demonstrate for the first time that acute GPR40 receptor activation leads to potentiation of GSIS in vivo and improves glucose tolerance even in insulin-resistant condition in rats. Taken together, these results suggest that GPR40 receptor agonists could be potential therapeutic alternatives to sulfonylureas. PMID- 19217449 TI - Parental history of type 2 diabetes mellitus, metabolic syndrome, and cardiometabolic risk factors in Asian Indian adolescents. AB - The objective was to study the influence of parental history of type 2 diabetes mellitus on prevalence of the metabolic syndrome (MS) and other cardiometabolic risk factors in Asian Indian adolescents. Adolescents aged 12 to 19 years (N = 321) were recruited from the Chennai Urban Rural Epidemiology Study. Based on parental diabetic status, 3 groups were studied: group 1, offspring of parents with normal glucose tolerance (n = 105); group 2, offspring of 1 diabetic parent (n = 114); and group 3, offspring of 2 diabetic parents (n = 102). Subjects underwent blood pressure and anthropometric measurements as well as an oral glucose tolerance test and a fasting lipid profile. Metabolic syndrome was diagnosed using the International Diabetes Federation definition. Body mass index (P < .001) and waist and hip circumference (P < .05 for group 2 and P < .001 for group 3) were significantly higher in groups 2 and 3 compared with group 1. High density lipoprotein cholesterol was significantly lower in groups 2 and 3 compared with group 1 (P < .05). Serum triglycerides were significantly higher in group 3 (P < .05) compared with the other 2 groups. Adolescents in group 3 (P < .001) and group 2 (P < .05) were significantly more overweight and had more abdominal obesity compared with those in group 1. Impaired fasting glucose and impaired glucose tolerance were also significantly higher in group 3 compared with the other 2 groups. High blood pressure showed an increasing trend from group 1 to group 3 (P for trend < .05). Two metabolic abnormalities were present in 7.6%, 14.9%, and 22.5% of adolescents in groups 1, 2, and 3, respectively (trend chi(2): 9.04, P = .003). Prevalence of MS was higher in groups 2 and 3 compared with group 1 but did not reach statistical significance because of small numbers. The cardiometabolic profile of the parents was similar to that of the adolescents. Parental history of type 2 diabetes mellitus increases risk of not only glucose intolerance but also other cardiometabolic risk factors like overweight, low high-density lipoprotein cholesterol, and high blood pressure in Asian Indian adolescents. PMID- 19217450 TI - A study of acid phosphatase locus 1 in women with high fat content and normal body mass index. AB - De Lorenzo and coworkers have recently described a class of women with normal body mass index (BMI) and high fat content (normal weight obese syndrome [NWO]). This observation prompted us to study the possible role of acid phosphatase locus 1 (ACP(1)) in the differentiation of this special class of obese subjects. Acid phosphatase locus 1 is a polymorphic gene associated with severe obesity and with total cholesterol and triglycerides levels. The enzyme is composed by 2 isoforms- F and S--that have different biochemical properties and probably different functions. The sample study was composed of 130 white women from the population of Rome. Total fat mass and percentage of fat mass were measured by dual-energy x ray absorptiometry. Thirty-six women had a BMI less than 25 and percentage of fat mass greater than 30 (high fat, normal BMI [HFHB]), and 94 women showed a BMI greater than 25 and a percentage of fat mass greater than 30 (high fat, high BMI [HFHB]). In the whole sample, the proportion of low-activity ACP(1) genotypes (*A/*A and *B/*A) was higher than in controls. However, whereas HFNB showed a very high frequency of ACP(1) *A/*A genotype, high-fat, high-BMI women showed an increase of *B/*A genotype. These 2 genotypes differ in the concentration of F isoform and the F/S ratio, which are lower in ACP(1)*A/*A genotype than in ACP(1)*B/*A genotype. The genetic differentiation of the class of women with normal BMI and high fat content from the class showing a concordant level of the 2 parameters supports the hypothesis that HFNB class represents a special cluster of obese subjects not revealed by BMI evaluation. Because ACP(1) is present in adipocytes, the present observation suggests that F isoform may have a specific role in the regulation of quantity of adipose tissue. PMID- 19217451 TI - High-molecular-weight adiponectin is a predictor of progression to metabolic syndrome: a population-based 6-year follow-up study in Japanese men. AB - Adiponectin is an adipocyte-specific secretory protein, which possesses antidiabetic and antiatherosclerotic properties. Adiponectin exists as multimers in serum, and high-molecular-weight (HMW) adiponectin is particularly considered to be the active form of the protein. The objective of the present study was to examine whether decreased HMW adiponectin is a predictor of progression to metabolic syndrome during a 6-year follow-up period in Japanese men. The study subjects were 416 Japanese men without metabolic syndrome, aged 30 to 59 years at baseline, who had participated in annual health checkups in both 2000 and 2006. Low concentration of HMW adiponectin (< or =2.65 microg/mL) was associated with substantially higher hazard ratio of the progression to metabolic syndrome after adjustment for age and body mass index (hazard ratio, 1.561; 95% confidence interval, 1.051-2.292; P = .028). The number of subjects with the progression to metabolic syndrome in each tertile based on baseline HMW adiponectin concentration was significantly different among the 3 groups (HMW adiponectin: chi(2) = 7.473, P = .0238; total adiponectin: chi(2) = 4.477, P = .1066; HMW total adiponectin ratio: chi(2) = 1.676, P = .4325). It was suggested that decreased HMW adiponectin is a predictor of the progression to metabolic syndrome in a 6-year follow-up study of Japanese men. Furthermore, it was suggested longitudinally that measuring HMW adiponectin is efficient to predict the progression to metabolic syndrome compared with measuring total adiponectin or HMW-total adiponectin ratio. PMID- 19217453 TI - Association of elevated serum alanine aminotransferase with metabolic factors in obese children: sex-related analysis. AB - Alanine aminotransferase (ALT) elevations are considered a surrogate marker of nonalcoholic liver disease and predict later development of diabetes and metabolic syndrome in adults. The aim of the present study is to evaluate the prevalence of high ALT levels in obese children using updated and sex-related cutoff ALT value (ALT >30 IU/L for boys and >19 IU/L for girls). We also analyzed the association between ALT levels and metabolic factors in the 2 sexes. Three hundred fifty-eight obese children (168 boys and 190 girls; age range, 6-16 years) were studied. Inclusion criteria were as follows: obesity, defined by an individual body mass index (BMI) greater than or equal to the 95th percentile for age and sex; negativity of markers for viral hepatitis; and no alcohol consumption. Two hundred six nonobese children (92 boys and 114 girls; age range, 6-16 years) served as a control group for ALT levels. The percentage of obese children with elevated ALT was 36% in boys and 55% in girls. Obese boys with ALT greater than 30 IU/L showed higher mother's BMI (P < .025), BMI, waist circumference, insulin resistance evaluated with homeostasis model assessment (HOMA-IR) index (P < .0001, for all), and systolic and diastolic blood pressure (P < .025, for both) compared with those with ALT not exceeding 30 IU/L. The ALT levels correlated positively with mother's BMI, BMI, waist circumference, HOMA IR, triglycerides, and blood pressure. In linear regression analysis, waist circumference was the only independent factor associated with ALT level (beta = 0.370, t = 3,905, P < .0001). Obese girls with ALT greater than 19 IU/L exhibited lower age (P < .025) and higher triglycerides (P < .0001) than girls with ALT not exceeding 19 IU/L. The ALT levels correlated positively with triglycerides and HOMA-IR and negatively with age and Tanner stage. In linear regression analysis, ALT levels were independently associated only with triglycerides (beta = 0.330, t = 4.588, P < .0001). Our study shows that a high proportion of obese children present elevated ALT levels. This abnormality is associated in boys, more than in girls, with preclinical traits of the metabolic syndrome. The adoption of sex related cutoff of ALT levels is desirable also for the pediatric population. PMID- 19217452 TI - Altered pattern of cannabinoid type 1 receptor expression in adipose tissue of dysmetabolic and overweight patients. AB - In overweight patients (OW), the increased peripheral activity of the endocannabinoid system in visceral adipose tissue (VAT) may be mediated by cannabinoid type 1 (CB1) receptor expression. We determined whether CB1 receptor splice variants and messenger RNA (mRNA) levels in perirenal and subcutaneous adipose tissues are associated with obesity and metabolic syndrome (MetS). Gene expression with multiple-primers real-time polymerase chain reaction (TaqMan; Applied Biosystem, Weiterstadt, Germany) was performed to study VAT and paired subcutaneous adipose tissue (SAT) mRNA from 36 consecutive patients undergoing nephrectomy. Cannabinoid type 1A and CB1E mRNAs variants with the longer version of exon 4 were expressed. The CB1 expression in perirenal VAT significantly correlated with body mass index (BMI). Paired subcutaneous/perirenal samples from normal-weight patients (BMI < 25 kg/m(2)) showed higher CB1 expression in SAT (P = .002), whereas in OW (BMI > or = 25 kg/m(2)), the higher CB1 expression was in VAT (P = .038). In unpaired samples, SAT of normal-weight patients had significantly higher CB1 mRNA levels compared with SAT of OW, whereas higher CB1 expression (P = .009) was found in VAT of OW (n = 25). Overweight patients with increased visceral CB1 expression had higher waist circumference (P < .01), insulin (P < .01), and homeostasis model assessment index (P < .01). In addition, patients with the MetS (n = 22) showed higher CB1 expression in perirenal adipose tissues (P = .007). Visceral adipose CB1 expression correlated with BMI. Overweight patients and those with MetS showed a CB1 expression pattern supporting a CB1-mediated overactivity of the endocannabinoid system in human VAT. PMID- 19217454 TI - Relationship between changes in insulin sensitivity and associated cardiovascular disease risk factors in thiazolidinedione-treated, insulin-resistant, nondiabetic individuals: pioglitazone versus rosiglitazone. AB - This study compared the effects of administering rosiglitazone (RSG) vs pioglitazone (PIO) on cardiovascular disease risk factors in insulin-resistant, nondiabetic individuals with no apparent disease. Twenty-two nondiabetic, apparently healthy individuals, classified as being insulin resistant on the basis of a steady-state plasma glucose concentration of at least 10 mmol/L during the insulin suppression test, were treated with either RSG or PIO for 3 months. Measurements were made before and after drug treatment of weight; blood pressure; fasting and daylong glucose, insulin, and free fatty acid (FFA) levels; and lipid and lipoprotein concentrations. Insulin sensitivity (steady-state plasma glucose concentration) significantly improved in both treatment groups, associated with significant decreases in daylong plasma concentrations of glucose, insulin, and FFA. Diastolic blood pressure fell somewhat in both groups, and this change reached significance in those receiving PIO. Improvement in lipid metabolism was confined to the PIO-treated group, signified by a significant decrease in plasma triglyceride concentration, whereas triglyceride concentration did not decline in the RSG-treated group, and these individuals also had increases in total (P = .047) and low-density lipoprotein cholesterol (P = .07). In conclusion, RSG and PIO appear to have comparable abilities to improve insulin sensitivity and lower daylong glucose, insulin, and FFA concentrations in nondiabetic, insulin resistant individuals. However, despite these similarities, their effects on lipoprotein metabolism seem to be quite different, with beneficial effects confined to PIO-treated individuals. PMID- 19217455 TI - Antidiabetic effects of dipeptidyl peptidase-IV inhibitors and sulfonylureas in streptozotocin-nicotinamide-induced mildly diabetic mice. AB - The present study investigated the antidiabetic effects of the dipeptidyl peptidase (DPP)-IV inhibitors ASP8497 and vildagliptin, and the sulfonylureas glibenclamide and gliclazide in streptozotocin-nicotinamide-induced mildly diabetic mice. A single administration of ASP8497 and vildagliptin significantly improved glucose tolerance by increasing plasma insulin and glucagon-like peptide 1 levels. In addition, a single administration of glibenclamide and gliclazide also caused significant improvement in glucose tolerance with an accompanying increase in the plasma insulin level. Subsequently, the effects of a 1-week chronic daily dosing of DPP-IV inhibitors and sulfonylureas were investigated. All drugs significantly improved glucose tolerance on day 1 of chronic daily dosing. After 1 week of chronic daily dosing, the DPP-IV inhibitors caused a significant improvement in glucose tolerance similar to those observed on day 1 by increasing the plasma insulin and glucagon-like peptide-1 levels. In contrast, the sulfonylureas had no significant improving or insulinotropic effect. Furthermore, ASP8497 also had an antihyperglycemic effect and improved pancreatic histopathologic lesions in a 4-week chronic daily dosing study. These results suggest that chronic daily dosing of sulfonylureas had virtually no antidiabetic effects because of marked attenuation of the insulinotropic action in streptozotocin-nicotinamide-induced mildly diabetic mice. In contrast, the antidiabetic efficacy of DPP-IV inhibitors, including ASP8497, did not change even after chronic daily dosing; therefore, DPP-IV inhibitors are useful as a therapeutic agent for impaired glucose tolerance and type 2 diabetes mellitus. PMID- 19217456 TI - Effect of moderate alcoholic beverage consumption on insulin sensitivity in insulin-resistant, nondiabetic individuals. AB - Although moderate alcohol consumption has been associated with a decrease in plasma insulin concentrations, relatively few studies have been conducted to evaluate the effect of alcohol on insulin sensitivity, particularly in nondiabetic, insulin-resistant individuals. Because enhanced insulin sensitivity could contribute to the reported association between moderate alcohol consumption and reduced risk of heart disease and diabetes, we believed it is important to address this issue. Consequently, we evaluated the ability of moderate alcohol consumption to improve insulin sensitivity, as measured by determining the steady state plasma glucose (SSPG) concentration during the insulin suppression test, in 20 nondiabetic, insulin-resistant individuals. Measurements were made of SSPG, glucose, insulin, and lipoprotein concentrations before and after consuming 30 g of alcohol for 8 weeks, either as vodka (n = 9) or red wine (n = 11). The SSPG concentrations (insulin resistance) decreased by approximately 8% in the total group (P = .08), and high-density lipoprotein cholesterol concentration increased by a mean of 0.09 mmol/L (P = .02). Trends were similar in individuals who consumed vodka or red wine. Men tended to have greater decline in SSPG and increase in high-density lipoprotein cholesterol compared with women. There were no other metabolic changes in fasting plasma glucose, insulin, and triglyceride concentrations. These data demonstrate that 8 weeks of moderate alcohol consumption had minimal impact on enhancing insulin sensitivity in nondiabetic, insulin-resistant individuals, raising questions as to the role, if any, of improved insulin sensitivity in the purported clinical benefits associated with moderate alcohol consumption. PMID- 19217457 TI - Low plasma basic fibroblast growth factor is associated with laser photocoagulation treatment in adult type 2 diabetes mellitus from the Veterans Affairs Diabetes Trial. AB - Basic fibroblast growth factor (bFGF) is a potent endothelial cell mitogen that does not normally circulate. Yet plasma bFGF-like bioactivity was increased in association with persistent microalbuminuria and retinopathy in adult type 2 diabetes mellitus. In the present study, we tested whether plasma bFGF immunoreactivity (IR) could predict the need for laser treatment of diabetic retinopathy in a baseline subset of advanced type 2 diabetes mellitus from the Veterans Affairs Diabetes Trial (mean: age, 59 years; diabetes duration, 11 years; baseline glycosylated hemoglobin, 9.5%). Plasma bFGF-IR was determined with a sensitive and specific 2-site enzyme-linked immunoassay in 172 patients at the baseline visit. Results were dichotomized at 4.5 pg/mL, the upper limit in healthy men. There was an unexpected significant association between low baseline plasma bFGF-IR level and the interim (4 years) need for laser treatment. First laser treatment was significantly more likely to be required in patients with low compared with high baseline bFGF (19% vs 6%, P = .03 for the difference). After adjusting for clinical risk factors, low vs high bFGF (hazard ratio [HR], 5.01; P = .012), duration of diabetes (HR, 1.05; P = .050), and low-density lipoprotein cholesterol concentration (HR, 0.98; P = .027) were all significantly associated with time to first laser occurrence. These and our prior results suggest that low plasma bFGF-IR may be a marker for the presence of anti-endothelial cell autoantibodies that may contribute to the need for laser photocoagulation treatment in adult men with advanced type 2 diabetes mellitus. PMID- 19217458 TI - The metabolism of plant sterols is disturbed in postmenopausal women with coronary artery disease. AB - In postmenopausal coronary artery disease (CAD) women, serum plant sterols are elevated. Thus, we investigated further whether serum plant sterols reflect absolute cholesterol metabolism in CAD as in other populations and whether the ABCG5 and ABCG8 genes, associated with plant sterol metabolism, were related to the risk of CAD. In free-living postmenopausal women with (n = 47) and without (n = 62) CAD, serum noncholesterol sterols including plant sterols were analyzed with gas-liquid chromatography, cholesterol absorption with peroral isotopes, absolute cholesterol synthesis with sterol balance technique, and bile acid synthesis with quantitating fecal bile acids. In CAD women, serum plant sterol ratios to cholesterol were 21% to 26% (P < .05) higher than in controls despite similar cholesterol absorption efficiency. Absolute cholesterol and bile acid synthesis were reduced. Only in controls were serum plant sterols related to cholesterol absorption (eg, sitosterol; in controls: r = 0.533, P < .001; in CAD: r = 0.296, P = not significant). However, even in CAD women, serum lathosterol (relative synthesis marker) and lathosterol-cholestanol (relative synthesis absorption marker) were related to absolute synthesis and absorption percentage (P range from .05 to <.001) similarly to controls. Frequencies of the common polymorphisms of ABCG5 and ABCG8 genes did not differ between coronary and control women. In conclusion, plant sterol metabolism is disturbed in CAD women; so serum plant sterols only tended to reflect absolute cholesterol absorption. Other relative markers of cholesterol metabolism were related to the absolute ones in both groups. ABCG5 and ABCG8 genes were not associated with the risk of CAD. PMID- 19217459 TI - Changes in insulin sensitivity, renal function, and markers of endothelial dysfunction in hypertension--the impact of microalbuminuria: a 13-year follow-up study. AB - Microalbuminuria (MA) clusters with the metabolic syndrome and insulin resistance and may reflect endothelial dysfunction. Microalbuminuria may also represent renal dysfunction. The aim of the present follow-up study was to assess changes over 13 years in insulin sensitivity, markers of endothelial dysfunction, and renal function in hypertensive subjects with and without MA in 1992-1993, matched for age, sex, and body mass index. Fourteen subjects with and 17 without MA at baseline (1992-1993) participated. At follow-up (2005-2006), MA status was unchanged in 75% of the subjects. The groups had comparable age, blood pressure, body mass index, markers of endothelial dysfunction, and metabolic traits, assessed by oral glucose tolerance test and hyperglycemic clamp. Estimated glomerular filtration rate decreased significantly in the MA group (P = .049) and tended to be lower in the MA than the non-MA group in 2005-2006 (79.9 +/- 24.5 vs 90.8 +/- 13.3 mL min(-1) (1.73 m(2))(-1), P = .2). Urinary albumin excretion in 1992-1993 predicted estimated glomerular filtration rate in 2005-2006 in adjusted analysis (beta = -0.47, P = .006). Estimated glomerular filtration rate less than 60 mL min(-1) (1.73 m(2))(-1) was more frequent in the MA than non-MA group at follow-up (P = .03). In conclusion, long-standing MA was not associated with progression of metabolic disturbances or markers of endothelial dysfunction in hypertensive individuals. A decline in renal function predicted by urinary albumin excretion was suggested. Microalbuminuria may not be a metabolic trait, but a marker mainly of renal endothelial dysfunction. PMID- 19217460 TI - Metabolic syndrome abating the beneficial effect of pravastatin treatment on adhesion of endothelium by monocytes in subjects with hypercholesterolemia. AB - Statin, a potent lipid-lowering agent, ameliorates the interaction of monocytes and endothelium, a critical step in the atherosclerotic process. However, it remains unclear whether this effect of statin depends on different doses or the presence of metabolic syndrome. In this prospectively double-blind study, 21 hypercholesterolemia subjects, with low-density lipoprotein cholesterol between 130 and 170 mg/dL, received low-dose (10 mg/d) or high-dose (40 mg/d) pravastatin treatment for 8 weeks. We assessed the reduction of monocyte adhesion to cultured endothelium between different-dose groups and the relationship to metabolic syndrome. Total cholesterol and low-density lipoprotein cholesterol were significantly decreased after 40-mg pravastatin treatment (-23.3% +/- 3.7%, P < .001 and -28.8% +/- 3.0%, P < .001), and the reductions were greater than those in the 10-mg group (P = .041 and P = .045, respectively). There was no significant difference in monocyte adhesion between high-dose and low-dose pravastatin treatment. When all subjects were divided into an improvement group and a no improvement group, according to the median of change percentage of monocyte adhesion after pravastatin treatment, there were significantly more subjects with metabolic syndrome in the no improvement than the improvement group (6 vs 1 person, P =.024). Using logistic regression analysis, metabolic syndrome, rather than dose effect of pravastatin, was an independent predictor of interaction between monocytes and endothelium (95% confidence interval = 0.001 0.865, P = .041). Attenuating adhesion between monocytes and endothelium is altered by the presence of metabolic syndrome when hypercholesterolemia subjects receive pravastatin treatment. PMID- 19217461 TI - Independent determinants of soluble form of receptor for advanced glycation end products in elderly hypertensive patients. AB - Advanced glycation end product receptor (RAGE) interaction plays an important role in atherosclerosis. Although exogenously administered soluble form of RAGE (sRAGE) has been shown to suppress the development and progression of atherosclerosis in animals, the kinetics and role of endogenous sRAGE in humans are not fully understood. In this study, to clarify whether endogenous sRAGE could capture and efficiently eliminate RAGE ligands such as circulating AGEs and high-mobility group box-1 (HMGB-1), we investigated the correlation between sRAGE and RAGE ligands and examined independent determinants of serum levels of sRAGE in hypertensive humans. Two-hundred seventy-one consecutive nondiabetic outpatients with essential hypertension (83 male and 188 female; mean age, 76.5 +/- 9.2 years) underwent a complete history, physical examination, and determination of blood chemistries, including serum levels of sRAGE, AGEs, and HMGB-1. Univariate regression analysis showed that serum levels of sRAGE were associated with body mass index (r = -0.313, P < .0001), waist (r = -0.214, P < .0001), alanine aminotransferase (r = -0.172, P = .005), gamma glutamyltranspeptidase (r = -0.213, P < .0001), 24-hour creatinine clearance (r = -0.348, P < .0001), B-type natriuretic peptide (r = 0.138, P = .027), tumor necrosis factor-alpha (r = 0.138, P = .002), and alcohol intake (r = -0.155, P = .010). By the use of multiple stepwise regression analyses, 24-hour creatinine clearance (P < .0001), gamma-glutamyltranspeptidase (P < .001), body mass index (P = .007), and tumor necrosis factor-alpha (P = .024) remained significant independently. The present study demonstrated for the first time that there was no significant correlation between serum levels of sRAGE and RAGE ligands such as circulating AGEs and HMGB-1 in hypertensive patients. Anthropometric and inflammatory variables and liver and renal function may be the determinants of endogenous sRAGE levels in nondiabetic hypertensive patients. PMID- 19217462 TI - Vascular complications in diabetic pregnancy. AB - Progression of diabetic retinopathy (DR) occurs at least temporarily during pregnancy and postpartum. Nephropathy is associated with increased risk of preeclampsia, nephrotic syndrome, preterm delivery, fetal growth restriction, and perinatal mortality. The degree of renal impairment and proteinuria in early pregnancy predict pregnancy complications in women with diabetic nephropathy. The main pregnancy complication in this respect is pre-eclampsia which increases with severity of proteinuria. Presence of retinopathy increases also risk of preeclampsia and also poor glycemic control. The pregnancy itself (first or subsequent) is not a long-term risk factor for developing microalbuminuria, any retinopathy, proliferative retinopathy, or neuropathy. These findings have practical implications for counselling young women in planning their pregnancies. Good glycemic control, normotension, lack of nephropathy as well as lack of pre proliferative/proliferative changes of diabetic retinopathy are good prognostic factors as regards the progression of vascular complications during pregnancy. Women with Type 1 diabetes should be evaluated before pregnancy for microangiopathies, treated and followed closely during pregnancy by obstetrician and internist/diabetologue. PMID- 19217463 TI - Predicting recurrences or major bleeding in women with cancer and venous thromboembolism. Findings from the RIETE Registry. AB - BACKGROUND: Cancer patients with venous thromboembolism (VTE) have an increased incidence of recurrences and bleeding complications Reliable information on the factors determining the risk for such complications may facilitate better use of therapy. METHODS: RIETE Registry is an ongoing, international registry of consecutive patients presenting with symptomatic acute VTE confirmed by objective tests. We assessed the 3-month outcome in all women with active cancer, trying to identify if differences exist according to the tumor site. RESULTS: Up to May 2007, 18,883 patients had been enrolled. Of them, 3805 (20%) had active cancer, 1719 (45%) were women. During the 3-month study period, 40 (2.3%) had recurrent deep vein thrombosis, 39 (2.3%) recurrent pulmonary embolism (PE), 67 (3.9%) major bleeding, 394 (23%) died. Of these, 13 (33%) women with recurrent PE died of the PE, 17 (42%) with major bleeding had fatal bleeding. In women with gastrointestinal (5.7% vs. 4.3%) or genitourinary (6.4% vs. 4.7%) cancers the incidence of bleeding complications exceeded that of VTE recurrences, while in those with brain (3.4% vs. 13%) or lung cancer (2.6% vs. 11%) the rate of recurrences outweighed that of major bleeding. CONCLUSIONS: We identified significant differences in outcome according to the site of cancer that may help to identify those women with cancer and VTE at a higher risk for recurrences or major bleeding. PMID- 19217464 TI - Thrombophilia and placenta-mediated pregnancy complications: from the bench to bedside to policy. PMID- 19217465 TI - Thrombophilia and pregnancy loss: cause or association. PMID- 19217466 TI - North American and British guidelines for anti-thrombotic therapy: are we reaching consensus? PMID- 19217467 TI - Guidelines for bleeding disorders in women. AB - Women with bleeding disorders are disproportionately affected by reproductive tract bleeding and other related complications. Data on the management of women with bleeding disorders are hampered by a lack of randomized trials, case-control studies or even large case series. In the absence of strong evidence to direct practice, government agencies and hemophilia organizations have developed consensus guidelines. This paper summarizes existing recommendations from these guidelines. PMID- 19217468 TI - Medico-legal aspects of thrombophilia and women's health. PMID- 19217469 TI - Cancer and thrombosis in women - molecular mechanisms. AB - Both women and men with cancer are at increased risk for developing venous thromboembolism (VTE), a propensity that has been known for many years. Until recently it was assumed, however, that the association between cancer and thrombosis is an epiphenomenon - not causally related to the transforming malignant events. The pathophysiology of thrombosis in patients with cancer is complex involving multiple tumor-related and host-related factors. Several recent studies have provided strong evidence that activation of blood coagulation, perhaps most often mediated by tissue factor (TF)-rich microparticles (MPs), is linked directly to oncogene-induced malignant transformation. In addition, the development of VTE, either before or concurrent with the diagnosis of cancer, appears to predict an aggressive behavior of a tumor, and correlates with increased tumor angiogenesis and early onset of distant metastasis. The regulation of expression of TF in tumor cells is controlled at the molecular level by several oncogenes, as appears to be true for cyclooxygenase-2 (COX-2), an important regulator of platelet function and plasminogen activator inhibitor type 1 (PAI-1), an inhibitor of fibrinolysis. In addition, engagement of protease activated receptors (PARs) by the TF-factor VIIa complex, factor Xa and/or thrombin, have now been shown to be important for tumor growth, angiogenesis and metastasis. Targeting blood clotting reactions in cancer, therefore, may provide a unique approach to cancer treatment. PMID- 19217470 TI - Menorrhagia: treatment options. PMID- 19217471 TI - Obstetric haemorrhage. PMID- 19217472 TI - Bleeding issues in neonates, infants and young children. PMID- 19217473 TI - Diagnostic issues of VTE in pregnancy. AB - Venous thromboembolism is a major cause of maternal morbidity and mortality, and accurate diagnostic workup upon suspicion of deep vein thrombosis or pulmonary embolism in a pregnant woman is of utmost importance. The diagnostic repertoire for venous thromboembolism is, however, less well studied in pregnant women. The clinical assessment is influenced by common symptoms of pregnancy such as leg swelling or shortness of breath. The role of D-Dimer is limited, since - even during uncomplicated pregnancy - D-Dimer levels increase with gestational age. Preliminary data indicate that a normal D-Dimer in a healthy pregnant woman with a low clinical probability may exclude deep vein thrombosis. Compression ultrasonography and ventilation perfusion scanning or helical computed tomography are the imaging techniques of choice in a pregnant woman with suspected deep vein thrombosis or pulmonary embolism, respectively. The role of magnetic resonance imaging for the diagnosis of venous thromboembolism during pregnancy is uncertain and contraindications particularly to contrast media have to be considered. PMID- 19217474 TI - Pathogenesis and management of antiphospholipid syndrome. PMID- 19217475 TI - Thrombophilia and gestational VTE. PMID- 19217476 TI - Treatment of venous thromboembolism during pregnancy. AB - BACKGROUND: Acute VTE occurs with an incidence of 1-2/1,000 during pregnancy and is associated with an acute mortality of 1-2%. However, data on VTE treatment during pregnancy are sparse: Even though 50 cases occur daily in the EU and US, a recent review identified only 174 cases in the literature. ACUTE TREATMENT: Standard treatment is LMWH at therapeutic doses or APTT-adjusted UFH. LMWH is at least as safe and effective as UFH in non-pregnant patients. In pregnancy, LMWH is the preferred option, because it offers better bioavailability, fewer injections, superior safety regarding HIT and osteoporosis. LONG-TERM TREATMENT: VKA are contraindicated because of teratogenicity and thus heparin is used for secondary prevention. Since UFH requires therapeutic doses throughout pregnancy, carrying the risk of osteoporosis, LMWH is the drug of choice. In a recent review most patients were treated initially with LMWH, predominately with twice daily injections. Recurrent VTE occurred in 1.2%, bleeding in 1.7%, with no HIT. Whether the long-term dose of LMWH can be reduced remains unresolved: Intermediate dose LMWH has been used effectively in cancer patients, who - like pregnant women - continue to have a high pro-thrombotic burden after the initial phase. CONCLUSION: Even though acute VTE is not uncommon and represents a life threatening event during pregnancy, data are sparse, and prospective trial data are needed to answer open questions concerning treatment modalities. Nevertheless, it is evident that LMWH is the preferred option for treatment of VTE during pregnancy. PMID- 19217477 TI - Novel approaches in management of immune thrombocytopenia. AB - Platelets play the key role in primary haemostasis. The number of platelets that can be recruited for these first steps of haemostasis is important, as platelets maintain vascular integrity. Therefore severe thrombocytopenia is associated with a high risk for bleedings. The overall incidence of chronic autoimmune thrombocytopenia (cAITP) is 1 to 10 per 100 000 in industrialized countries. Like most other autoimmune disorders, cAITP is more common among adults, and thereby women are more often affected than man. It is relatively benign and rarely requires treatment. Furthermore, most treatment modalities are associated with severe adverse effects that may outweigh the benefits. Therefore, currently treatment is considered appropriate only for symptomatic patients and for those at high risk of bleeding, either because of a surgical procedure or because of very low platelet counts. Short-term treatment aims to increase platelet counts in patients with acute bleeding symptoms or in those undergoing surgery. Long term treatment may be considered to increase the quality of life, if patients are affected by low platelet counts in the form of bleeding or anxiety. Recently, two new substances have been shown to increase platelet counts in cAITP, romiplostim and eltrombopag. Both bind to the thrombopoietin receptor thereby inducing an increase of platelet formation. Remarkably, the rate of treatment success was independent of any stratification modalities like previous response to drugs or splenectomy, and was associated with rather few and mild adverse effects. Further, it was previously assumed that thrombopoiesis is already turned on to compensate increased platelet sequestration in cAITP. The treatment success shows that thrombopoiesis can be increased to enhance platelet production, strongly supporting impaired thrombopoiesis in cAITP. It is likely that romiplostim and eltrombopag will significantly add to the treatment armaments for bridging certain situations of increased risk of bleeding in cAITP, mainly if corticosteroids are contraindicated, or for non-responders to first-line treatment modalities. Their role in long-term treatment and in deferring splenectomy will be determined by their price and foremost by their proven safety in long-term observational studies. PMID- 19217478 TI - Thrombotic microangiopathy in pregnancy. PMID- 19217479 TI - Disseminated intravascular coagulation (DIC) in pregnancy and the peri-partum period. PMID- 19217480 TI - Epidemiology of hormone-related venous thromboembolism. PMID- 19217481 TI - Mechanisms of thrombosis related to hormone therapy. AB - Combined oral contraceptives and combined oral postmenopausal hormone therapy are associated with a weak, but clinically significant risk of arterial and venous thrombosis (VT). The effects are related to dose of estrogen and type of progestin. The main effects are increase in markers of activated coagulation, reduction in coagulation inhibitors, and acquired activated protein C resistance. Reduction in tissue factor pathway inhibitor (TFPI) is probably an important mechanism, which predicts activation of coagulation and acquired resistance to activated protein C. Coagulation markers should be used as intermediate or surrogate markers in early pharmacodynamic studies to evaluate the risk associated with new formulations. PMID- 19217482 TI - Counselling women about hormonal therapy. PMID- 19217483 TI - Thrombophilia and fetal loss: Lessons from gene targeting in mice. AB - Inherited thrombophilia is recognized as one of the causes of recurrent fetal loss. Yet, the risk of fetal loss in women with thrombophilia is far from absolute. Other risk modifiers are clearly involved, but remain to be identified. The mechanism that translates maternal thrombophilia into pregnancy disorder is also not understood. Genetically engineered mice have led to the development of new insights into the etiology and pathogenesis of thrombophilia-associated fetal loss. These insights are the focus of this review. PMID- 19217484 TI - Platelet activation impairs placental function. PMID- 19217485 TI - Microparticles and placental hemostasis. PMID- 19217486 TI - Angiogenic factors and preeclampsia. AB - Preeclampsia/eclampsia is a major cause of maternal and fetal morbidity worldwide. Although the etiology of preeclampsia is still unclear, the clinical phenotypes of preeclampsia have been demonstrated to be related to high circulating levels of anti-angiogenic proteins secreted by the placenta such as soluble Fms-like tyrosine kinase 1 (sFlt1) and soluble endoglin. Because, alterations in circulating sFlt1 and soluble endoglin precede the onset of clinical disease, these factors may be useful to screen or identify patients at risk for preeclampsia. Investigations are currently underway of various pharmacologic agents to counteract the effects of sFlt1 and/or sEng as a potential treatment for preeclampsia. Recently several isoforms of sFlt1 have been described, such as sFlt1-14 which is expressed only in primates, and is thought to be the primary isoform produced by the placenta in preeclamptic subjects. Although several novel pathways have been proposed to play key roles in inducing sFlt1 production, the exact role of these pathways in human preeclampsia is still not known. Women with a history of preeclampsia have an increased risk of hypertension, and cardiovascular and renal disease. Whether these long-term observations are due to persistent and subtle endothelial damage as result of preeclampsia, or simply reflect the consequences of the vascular risk factors which are more common in these women remains unknown. PMID- 19217487 TI - Disabled inside. PMID- 19217488 TI - Preface. Home care, from origins to present day. PMID- 19217489 TI - The past, present, and future of skilled home health agency care. AB - This article reviews the past history of home health agency care from its beginnings to the present day, evidence regarding the effect of recent changes in financing on these services, the state of skilled home health care in 2008, and a discussion of future directions. Home health care serves several million patients per year, many of whom are recuperating from acute illness episodes. Due to illness burden and Medicare funding, a large proportion of care that home health agencies deliver is geriatric care. However, home health care plays an important role for patients of all ages with significant acute and chronic illnesses. Medicare home health care suffered a significant downturn following the 1997 Balanced Budget Act and is recovering under Prospective Payment. Like most sectors of care, home health care has often operated in a "silo" but there is increasing recognition of the need to bridge care settings and provide care continuity for sick, chronically ill individuals. This is an important challenge for the future. Agencies that have strong information technology infrastructure and chronic care management systems along with a seasoned clinical workforce will be well positioned for key roles in home health care in decades to come. Home health care serves several million patients each year, many of whom are recuperating from acute illness episodes. Due to the burden of illness and Medicare funding, a large proportion of care that home health agencies deliver is geriatric care. However, home health care plays an important role for patients of all ages with significant acute and chronic illnesses. Medicare home health care suffered a significant downturn following the 1997 Balanced Budget Act (BBA) and is recovering under prospective payment. Like most sectors of care, home health care has often operated in a "silo," but there is increasing recognition of the need to bridge care settings and provide care continuity for sick, chronically ill individuals. This is an important challenge for the future. Agencies that have strong information technology infrastructure and chronic care management systems along with a seasoned clinical workforce will be well positioned for key roles in home health care in the decades to come. PMID- 19217490 TI - The past, present, and future of house calls. AB - This article describes: the origin and evolution of house calls; what is uniquely gained from house calls for both patients and providers; key clinical issues managed at home; evidence that house calls improve outcomes; organization of house call practices; and changes in Medicare reimbursement, technology, and interdisciplinary team care that have influenced the delivery of home-based medical care. PMID- 19217491 TI - Advances and issues in personal care. AB - The focus of this article is paid personal assistant service (PAS) workers in community settings, and the converging current and future trends affecting the need to strengthen and expand the PAS workforce. A number of ways in which the federal and state governments, and the private sector can begin to solve the short and long-term workforce challenges are highlighted. PMID- 19217492 TI - Elder abuse and neglect: when home is not safe. AB - The prevalence and seriousness of elder abuse and neglect require the collaboration of health care professionals with many other disciplines for adequate assessment and intervention. The home visit provides a unique opportunity for the visitor to evaluate risk factors. Interventions and reporting depend on available resources, expertise and local reporting laws. Possible reasons for low physician and victim self-reporting are reviewed. Domestic violence persists into late life and requires different approaches than dealing with caregiver burnout or self-neglect. Involvement of health professionals in educating others in the community about elder abuse and neglect may allow isolated at-risk elders to be identified. PMID- 19217493 TI - Assistive technologies in the home. AB - Assistive technologies are critical to elders maintaining independence in the home. Adequate assessment of the patient's needs, the appropriateness of the device to that need, and the patient's motivation to use of a device is required for successful outcomes. A team approach is needed to ensure that devices are correctly prescribed, and the patient is taught how to use it effectively. A wide range of devices is available to support activities of daily living, mobility, home management, and safety. The use of personal computers is significantly expanding the possibility of independent living through support systems, monitoring systems, and information resources. PMID- 19217494 TI - Hospital at home. AB - Although the acute hospital is the standard venue for treating acute serious illness, it is often a difficult environment for older adults who are highly susceptible to functional decline and other iatrogenic consequences of hospital care. Hospital care is also expensive. Providing acute hospital-level care at home, in lieu of usual institutional care, is viable. As an emerging service model, the definition of hospital at home (HaH) remains unsettled. Data favor HaH models that provide substantial physician inputs and are geared toward substituting for hospital care, provide service that is highly satisfying to patients and their caregivers, are associated with less iatrogenic complications, and are less expensive. Dissemination of HaH in integrated delivery systems is feasible. Widespread dissemination of HaH in the United States will require payment reform that acknowledges the role of HaH in the health care system. PMID- 19217495 TI - New diagnostic and information technology for mobile medical care. AB - Medicare reimbursement for home visits average around $100 without ancillaries, so making 10 home visits to prevent even a single $1,000 ambulance ride is cost neutral for Medicare. Home medical care is only an added cost if it fails to offset acute care use. The government's demographic and financial pressure suggests a need to press ahead with the enhanced mobile care model, so the explosion in point-of-care devices should continue. The main challenge is to decide which ones provide dispositive value to patients. PMID- 19217496 TI - Workforce development in geriatric home care. AB - With the rapidly aging population, it is anticipated that within two decades several million more individuals in the United States with functional impairment and serious ill health will need home health care. This article discusses workforce development, which is a critical issue for future planning, as recently highlighted by the Institute of Medicine (IOM). Key aspects of recruitment, training, and retention of home care workers are discussed, including those who provide basic support for activities of daily living as well as a variety of skilled professionals: therapists, nurses, pharmacists, and physicians. Although the geriatric workforce shortage affects all care settings, it is especially critical in home health care, in part because we are starting with far too few clinicians to meet the medical needs of homebound elderly. A combination of actions is needed, including educational programs, such as those developed by the American Academy of Home Care Physicians (AAHCP), changes in financial incentives, and changes in the culture and practice of health care, to make the home the primary focus of care for these vulnerable, underserved individuals rather than an afterthought. PMID- 19217497 TI - The history of quality measurement in home health care. AB - Quality improvement is as central to home health care as to any other field of health care. With the mandated addition in 2000 of Outcome Assessment and Information Set (OASIS) and outcome-based quality improvement (OBQI), Medicare home health agencies entered a new era of documenting, tracking, and systematically improving quality. OBQI is augmented by the Medicare Quality Improvement Organization (QIO) program, which is now entering the ninth in a series of work assignments, with the tenth scope in the planning stages. Evidence has shown that applied quality improvement methods can drive better outcomes using important metrics, such as acute care hospitalization. This article reviews key findings from the past 2 decades of home care quality improvement research and public policy advances, describes specific examples of local and regional programmatic approaches to quality improvement, and forecasts near-future trends in this vital arena of home health care. PMID- 19217498 TI - Care transitions and home health care. AB - Transitions of care are becoming recognized as an important area for improvement in health care quality and patient safety. Yet there remains consistent evidence from multiple studies in varied settings of failures to complete safe, effective hand-offs from one location of care to the next. Major lapses include absent or limited clinical information and care plan content, plus errors related to medications. There are identifiable problems with half or more of the transitions that occur between care settings, and adverse consequences occur in 15 to 25 percent of patients. Undoubtedly these lapses contribute to the rates of re hospitalization in post-acute care which affect 20 to 30 percent of patients within 60 days after hospital discharge. This article reviews models of transitional care intervention that have been tested and shown to be effective including less intensive coaching or guided care approaches, and more intensive case management strategies. Effective transitional care processes, linked with strong home care programs can reduce re-hospitalization by a third in some less intensive models and by half or more in some more intensive models. PMID- 19217499 TI - Veteran's Affairs Home Based Primary Care. AB - In response to the anticipated growth of the veteran population with chronic disabling diseases, the Department of Veterans Affairs (VA) established Home Based Primary Care (HBPC). This article focuses on that program, a home care program that specifically targets individuals with complex chronic disabling disease, with the goal of maximizing the independence of the patient and reducing preventable emergency room visits and hospitalizations. HBPC programs provide comprehensive longitudinal primary care by an interdisciplinary team in the homes of veterans with complex chronic disease, who are not effectively managed by routine clinic-based care. HBPC is very different from and complementary to standard skilled home care services, in population, processes and outcomes. HBPC targets persons with advanced chronic disease, rather than remediable conditions. HBPC provides comprehensive care of multiple co-morbidities, rather than problem focused care. HBPC is delivered by an interdisciplinary team, rather than one or two independent providers. Currently operating in three-fourths of VA facilities, HBPC expansion continues to be driven by clinical success and the highest satisfaction of all VA services. VA HBPC is a model to emulate for the care of persons with complex, chronic disabling conditions, improving quality without added cost, and maximizing their independence through comprehensive longitudinal interdisciplinary care delivered in their homes. PMID- 19217500 TI - Independence at home: community-based care for older adults with severe chronic illness. AB - By most clinical and economic measures, our health care system is not providing effective or affordable care to Medicare beneficiaries with severe chronic illness. Two million elders, constituting most of the 5% who account for nearly half of Medicare costs, have multiple chronic conditions, functional disability, and average per capita costs of over $50,000 per year. Prior reforms aimed at this population did not change the flawed delivery system, which remains centered in the doctor's office, hospitals, and nursing homes. This article describes a model of coordinated home-based medical care, called Independence at Home (IAH), which operates on a limited basis in many US communities and in the Veterans Affairs system. IAH-type teams deliver a full range of medical and social services at home to seriously ill elders and thereby reduce overall health care costs. We review the evidence that this approach can lower total costs by 25 percent or more while improving patient satisfaction and outcomes. We discuss funding for the new model, which also produces net savings for Medicare. A Medicare reform bill, called the Independence at Home Act, was introduced in the US House and Senate in 2008 to promote replication of this mobile elder care model. PMID- 19217501 TI - Introduction: gastrointestinal (GI) malignancies. PMID- 19217502 TI - Review of the gastrointestinal tract: from macro to micro. AB - OBJECTIVES: To review the normal anatomy and physiology of the gastrointestinal (GI) tract, the malignant transformations in GI cancers, and the rationale for targeted therapy for these cancers. DATA SOURCES: Published articles, book chapters and web sources. CONCLUSION: Oncology nurses require an understanding of normal GI anatomy and physiology, along with an understanding of malignant transformations at the cellular and molecular level, to effectively educate and care for the patient with a diagnosis of a GI cancer. IMPLICATIONS FOR NURSING PRACTICE: Challenges for the oncology nurse include continuing education related to GI cancer, the development of effective patient education skills, ensuring safe administration of oral agents and remaining current regarding GI clinical trial opportunities. Education of nursing colleagues, development of an area of expertise through specialization, and development of leadership skills are opportunities associated with practicing in the dynamic environment of oncology nursing. PMID- 19217503 TI - Prevention and screening of gastrointestinal cancers. AB - OBJECTIVES: This article reviews the current recommendations and data supporting various screening and prevention strategies for colorectal cancer (CRC) in average and high-risk individuals. DATA SOURCES: Published research reports, epidemiologic data, and published guidelines from professional organizations. CONCLUSION: Properly applied screening tools can potentially decrease the morbidity and mortality associated with CRC. IMPLICATIONS FOR NURSING PRACTICE: Nurses need to be aware of current recommendations for the early detection of CRC so they can provide patients with an accurate assessment of risk for developing CRC and education about the appropriate CRC screening guidelines. PMID- 19217504 TI - Colon, rectal, and anal cancers. AB - OBJECTIVES: To review the incidence, risk factors, staging, diagnosis, and treatment of colon, rectal, and anal cancers, as well as nursing care associated with managing patients diagnosed with these malignancies. DATA SOURCES: Published research reports, epidemiologic data, published patient management guidelines, and institution-based clinical tools. CONCLUSIONS: Significant advances in the management of colon, rectal, and anal cancers in the past decade have extended patient survival. Further clinical research will refine current therapeutic strategies and treatment decision-making aids while minimizing symptoms of disease and treatment. IMPLICATIONS FOR NURSING PRACTICE: Nurses need to be familiar with risk factors, disease course, and current and emerging therapies to assist patients with treatment decision-making, and to anticipate and intervene in managing disease and treatment-induced problems. Early identification and management of distressing symptoms can help to avoid life-threatening effects and promote patient adherence to prescribed therapies; timely patient/family education may minimize anxiety and promote self-management. PMID- 19217505 TI - Gastrointestinal stromal tumors and neuroendocrine tumors. AB - OBJECTIVES: Patients with rare gastrointestinal (GI) malignancies can exhibit unique objective and subjective manifestations. This article is a primer for the fundamental understanding of some of these diseases, namely gastrointestinal stromal tumors (GIST) and gastroenteropancreatic neuroendocrine tumors (NET) and therapeutic strategies. DATA SOURCES: Epidemiologic data, published research reports, national guidelines for oncology practice, and personal experience. CONCLUSION: Despite the rarity of GIST, gastroenteropancreatic neuroendocrine tumors, gastric lymphoma, and adenocarcinoma of the small bowel, oncology nurses must be prepared to effectively assess, plan, and implement care strategies for these patients. IMPLICATIONS FOR NURSING PRACTICE: Caring for patients with uncommon GI malignancies is challenging for oncology nurses whose experience with these tumors is limited. Fundamental knowledge and awareness of resources can help to ensure optimal patient care. Case vignettes illustrate patient presentation and formulation of treatment recommendations. PMID- 19217506 TI - Tumors of the esophagus, gastroesophageal junction, and stomach. AB - OBJECTIVES: To review the incidence, risk factors, staging, and diagnosis of esophageal, gastroesophageal junction, and stomach cancers, as well as nursing challenges associated with managing patients with these tumors. DATA SOURCES: Published research reports, epidemiologic data, published patient management guidelines, and institution-based clinical tools. CONCLUSION: While survival is slowly increasing, morbidity and mortality associated with upper gastrointestinal (UGI) cancers remain high. Research is needed to identify therapeutic approaches that will effectively decrease tumor burden and minimize physical consequences associated with disease and treatment. IMPLICATIONS FOR NURSING PRACTICE: Nurses need to be familiar with risk factors, disease course, and future therapies of UGI cancers to provide education relevant to preventive approaches, assist patients with treatment decision-making, and manage disease and treatment-induced sequela. Early identification and management of postoperative complications and nutritional issues can avoid life-threatening effects. PMID- 19217507 TI - Cancers of the pancreas and hepatobiliary system. AB - OBJECTIVES: To review the incidence, risk factors, staging, and diagnosis of hepatobiliary malignancies and pancreas cancer as well as nursing challenges associated with managing patients with these tumors. DATA SOURCES: Published research reports, epidemiologic data, patient management guidelines, and institution-based clinical tools. CONCLUSION: While survival is slowly increasing, morbidity and mortality associated with hepatobiliary and pancreas cancers remains high. Further scientific study is needed to identify methods of detecting these malignancies earlier and to develop therapeutic approaches that will effectively decrease tumor burden while simultaneously palliating disease and treatment-induced consequences including both physical and emotional effects. IMPLICATIONS FOR NURSING PRACTICE: Nurses need to be familiar with the natural history of these diseases and current and future therapies to facilitate treatment decision-making when options are limited, anticipate and intervene in managing disease and treatment-induced effects, and support clinical research efforts. PMID- 19217508 TI - Basic aspects and clinical applicability in the field of lipidology. Foreword. PMID- 19217509 TI - The field of clinical lipidology. Preface. PMID- 19217510 TI - Advanced lipoprotein testing: recommendations based on current evidence. AB - Low-density lipoprotein cholesterol (LDL-C) level currently is used as the major determinant of lipid- and lipoprotein-associated risk for ischemic cardiovascular disease, and varying levels have become the standard goals of lipid-altering treatment. The predictive value of the LDL-C cholesterol level, however, often is less than that provided by other variables such as non-high-density lipoprotein cholesterol (non-HDL-C), apolipoprotein B (apoB), and the number of LDL particles measured by nuclear magnetic resonance spectroscopy. This article reviews studies that compare these different lipoprotein variables, describes advanced methodologies of lipoprotein testing, and suggests goals of treatment and clinical situations in which these tests might be ordered. PMID- 19217511 TI - Risk scores for prediction of coronary heart disease: an update. AB - Risk scores for the prediction of coronary heart disease (CHD) have greatly improved in the past 30 years. While standardized baseline measurements and modern technology aid in the development of increasingly accurate CHD risk algorithms, recent reports have shown that simple prediction tools using a basic set of variables, including age, systolic blood pressure, smoking, hypertension, exercise, body mass index, diabetes, and family history are predictive of CHD risk and can potentially be self-administered. PMID- 19217512 TI - Lifestyle approaches and dietary strategies to lower LDL-cholesterol and triglycerides and raise HDL-cholesterol. AB - This article discusses specific dietary factors as well as dietary patterns that affect the major coronary heart disease (CHD) lipid risk factors (ie, LDL-C, HDL C, and TG). Based on a very large evidence base, it is clear that diet and lifestyle practices can markedly affect these major CHD lipid risk factors, and consequently decrease CHD risk substantively. PMID- 19217513 TI - Lowering low-density lipoprotein cholesterol: statins, ezetimibe, bile acid sequestrants, and combinations: comparative efficacy and safety. AB - Statins, ezetimibe, and bile acid-binding resins can be used individually or in combination for lowering low-density lipoprotein cholesterol (LDL-C) levels. Statins are the most potent drugs for lowering LDL-C and are well tolerated in most patients. The addition of a bile acid sequestrant or ezetimibe to a statin produces additional LDL-C reduction allowing many patients to reach LDL-C targets. This article discusses the efficacy and safety of available statins, bile acid sequestrants, and ezetimibe in the treatment of hyperlipidemia. PMID- 19217514 TI - Other therapies for reducing low-density lipoprotein cholesterol: medications in development. AB - Although the past 30 years have been fruitful and productive in lipid research, from basic science to drug development to demonstration of clinical benefit, cardiovascular disease remains the major cause of mortality and morbidity in industrialized societies. With the rapid industrialization of countries, such as India and China, cardiovascular disease rapidly is becoming the leading cause of global death and disability. Although most of the effective lipid-lowering drugs, the statins, have become generic and inexpensive, there remains a need for effective and safe agents. Hopefully, some of those discussed in this article will fill that need. PMID- 19217515 TI - Managing statin myopathy. AB - Approximately 10% of patients treated with statins experience some form of muscle related side effects in clinical practice. These can range from asymptomatic creatine kinase (CK) elevation, to muscle pain, weakness, and its most severe form, rhabdomyolysis. Higher risk patients for statin myopathy are those older than 80, with a small body frame, on higher statin doses, on other medications, or with other systemic diseases including hepatic or renal diseases, diabetes mellitus, or hypothyroidism. The cause of statin myopathy is presumed to be the same for its variable presentation but has not been defined. In patients with myopathic symptoms, their symptoms and CK levels determine whether statin therapy can be continued or must be stopped. PMID- 19217516 TI - Hypertriglyceridemia: impact and treatment. AB - The treatment of elevated levels of low-density lipoprotein cholesterol is standard medical practice supported by conclusive outcome data. Less definitive information exists for hypertriglyceridemia. Only in the setting of severe hyperchylomicronemia is the benefit of triglyceride lowering clear: it is a means to reduce the risk of pancreatitis. The relationship of triglycerides and cardiovascular disease is still unclear. Moreover, the cardiovascular benefits of reducing triglycerides and of using triglyceride-lowering medications remain unproved. Nonetheless it has become almost standard to reduce the levels of triglyceride-rich lipoproteins that are a major component of plasma non-high density lipoprotein cholesterol. PMID- 19217517 TI - Novel therapies for increasing serum levels of HDL. AB - The protectiveness of elevated HDL-C against CHD and its long-term sequelae is a subject of intense investigation throughout the world. HDL has the capacity to modulate a large number of atherogenic mechanisms, such as inflammation, oxidation, thrombosis, and cell proliferation. Among lipoproteins, HDL is also unique, in that it promotes the mobilization and clearance of excess lipid via the series of reactions collectively termed "reverse cholesterol transport." Numerous therapeutic agents are being developed in an attempt to modulate serum levels of HDL-C as well as its functionality. This article discusses the development of newer treatments targeted at raising HDL-C and HDL particle numbers to reduce residual risk in patients at risk for CHD. PMID- 19217518 TI - Lipid management in children. AB - Atherosclerosis begins in childhood, and early initiation of prevention through behavioral means may lower the risk of future cardiovascular disease. The obesity epidemic threatens the cardiovascular health of today's children. Genetic dyslipidemias such as familial hypercholesterolemia and the presence of multiple risk factors in the same child or adolescent may require pharmacologic therapy. PMID- 19217519 TI - Lipid management in the geriatric patient. AB - Elderly individuals are at higher risk for cardiovascular events, and thus this population stands to gain a greater reduction in events from lipid therapy than younger individuals. Multiple primary and secondary prevention trials have demonstrated that the benefits of statins in geriatric patients are equivalent to, or greater than, those seen in younger patients. Combination therapy with non statin agents should be considered in patients who do not meet cholesterol goals or who have concomitant hypertriglyceridemia or low levels of high-density lipoprotein cholesterol. Although increased side effects may occur with high-dose statin therapy, careful vigilance of drug interactions and limiting polypharmacy can reduce these effects. PMID- 19217520 TI - Lipid management in patients who have HIV and are receiving HIV therapy. AB - Dyslipidemia now is recognized as a significant potential adverse event in HIV infected patients who are receiving antiretroviral therapy. HIV-infected persons who have hyperlipidemia should be managed similarly to those without HIV infection in accordance with the National Cholesterol Education Program. Providers must treat the HIV infection first; if dyslipidemia develops, patients should be prescribed lipid-lowering therapies or should consider modifying their current antiretroviral therapy, if indicated. Evidence for these two strategies is discussed. PMID- 19217521 TI - Lipid management in chronic kidney disease, hemodialysis, and transplantation. AB - Recent studies have shown the spectrum of dyslipidemia in patients who have chronic kidney disease (CKD) or end-stage renal disease to be different from that of the general population. This article discusses the pathophysiology of dyslipidemia in CKD, dialysis, and renal transplant patients, the therapeutic options, and their association with clinical outcomes. Whenever possible, comparisons are made to outcomes in the general population. PMID- 19217522 TI - (18)F-Labeled benzylideneaniline derivatives as new ligands for beta-amyloid plaque imaging in Alzheimer's disease. AB - INTRODUCTION: Noninvasive early detection of beta-amyloid (Abeta) plaques might be useful for the treatment of patients with Alzheimer's disease (AD). We herein describe the synthesis of (18)F-labeled benzylideneaniline derivatives using a novel labeling approach for imaging Abeta plaques in AD patients. METHODS: Benzylidenaniline derivatives were synthesized by reacting fluorobenzaldehyde and corresponding aniline derivatives. Fluorobenzaldehyde was labeled with (18)F by incubating [(18)F]fluoride with N,N,N-trimethylbenzaldehyde in the presence of tetrabutylammonium bicarbonate. In vitro binding assay, stability test and biodistribution study were performed. RESULTS: These compounds were stable at alkaline pH (pH >9); however, they were hydrolyzed rapidly at physiological pH (pH approximately 7.4). The K(i) values of amine-containing benzylideneaniline derivatives for Abeta(1-40) and Abeta(1-42) aggregates were 26-78 nM. These (18)F labeled benzylideneaniline derivatives showed high brain uptake and rapid clearance after intravenous administration in normal mice (1.8-3.1%ID/g at 2 min and 0.1-1.2%ID/g at 30 min). The low level of bone activity at 30 min indicated that these (18)F-labeled benzylideneanilines are not prone to defluorination. Furthermore, the compounds have suitable lipophilicity - a property required to penetrate the blood-brain barrier. CONCLUSION: These results showed that the instability of these compounds could cause a higher early phase/late phase ratio due to rapid clearance in the normal brain. The findings from this study suggest that these (18)F-labeled benzylideneaniline derivatives are feasible for the imaging of Abeta plaques. PMID- 19217523 TI - Labeling internalizing anti-epidermal growth factor receptor variant III monoclonal antibody with (177)Lu: in vitro comparison of acyclic and macrocyclic ligands. AB - INTRODUCTION: The monoclonal antibody (mAb) L8A4, reactive with the epidermal growth factor receptor variant III (EGFRvIII), internalizes rapidly in glioma cells after receptor binding. Combining this tumor-specific mAb with the low energy beta-emitter (177)Lu would be an attractive approach for brain tumor radioimmunotherapy, provided that trapping of the radionuclide in tumor cells after mAb intracellular processing could be maximized. MATERIALS AND METHODS: L8A4 mAb was labeled with (177)Lu using the acyclic ligands [(R)-2-amino-3-(4 isothiocyanatophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diamine-pentaacetic acid (CHX-A''-DTPA), 2-(4-isothiocyanatobenzyl)-diethylenetriaminepenta-acetic acid (pSCN-Bz-DTPA) and 2-(4-isothiocyanatobenzyl)-6 methyldiethylenetriaminepentaacetic acid (1B4M-DTPA), and the macrocyclic ligands S-2-(4-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-tetraacetic acid (C DOTA) and alpha-(5-isothiocyanato-2-methoxyphenyl)-1,4,7,10-tetraazacyclododecane 1,4,7,10-tetraacetic acid (MeO-DOTA). Paired-label internalization and cellular processing assays were performed on EGFRvIII-expressing U87.DeltaEGFR glioma cells over 24 h to directly compare (177)Lu-labeled L8A4 to L8A4 labeled with (125)I using either iodogen or N-succinimidyl 4-guanidinomethyl-3 [(125)I]iodobenzoate ([(125)I]SGMIB). In order to facilitate comparison of labeling methods, the primary parameter evaluated was the ratio of (177)Lu to (125)I activity retained in U87.DeltaEGFR cells. RESULTS: All chelates demonstrated higher retention of internalized activity compared with mAb labeled using iodogen, with (177)Lu/(125)I ratios of >20 observed for the three DTPA chelates at 24 h. When compared to L8A4 labeled using SGMIB, except for MeO-DOTA, internalized activity for (125)I was higher than (177)Lu from 1-8 h with the opposite behavior observed thereafter. At 24 h, (177)Lu/(125)I ratios were between 1.5 and 3, with higher values observed for the three DTPA chelates. CONCLUSIONS: The nature of the chelate used to label this internalizing mAb with (177)Lu influenced intracellular retention in vitro, although at early time points, only MeO-DOTA provided more favorable results than radioiodination of the mAb via SGMIB. PMID- 19217524 TI - Development of [(90)Y]DOTA-conjugated bisphosphonate for treatment of painful bone metastases. AB - INTRODUCTION: Based on the concept of bifunctional radiopharmaceuticals, we have previously developed (186)Re-complex-conjugated bisphosphonate analogs for palliation of painful bone metastases and have demonstrated the utility of these compounds. By applying a similar concept, we hypothesized that a bone-specific directed (90)Y-labeled radiopharmaceutical could be developed. METHODS: In this study, 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) was chosen as the chelating site, and DOTA was conjugated with 4-amino-1-hydroxybutylidene 1,1-bisphosphonate. [(90)Y]DOTA-complex-conjugated bisphosphonate ([(90)Y]DOTA HBP) was prepared by coordination with (90)Y, and its biodistribution was studied in comparison to [(90)Y]citrate. RESULTS: In biodistribution experiments, [(90)Y]DOTA-HBP and [(90)Y]citrate rapidly accumulated and resided in the bone. Although [(90)Y]citrate showed a higher level of accumulation in the bone than [(90)Y]DOTA-HBP, the clearances of [(90)Y]DOTA-HBP from the blood and from almost all soft tissues were much faster than those of [(90)Y]citrate. As a result, the estimated absorbed dose ratios of soft tissues to osteogenic cells (target organ) of [(90)Y]DOTA-HBP were lower than those of [(90)Y]citrate. CONCLUSIONS: [(90)Y]DOTA-HBP showed superior biodistribution characteristics as a bone-seeking agent and led to a decrease in the level of unnecessary radiation compared to [(90)Y]citrate. Since the DOTA ligand forms a stable complex not only with (90)Y but also with lutetium ((177)Lu), indium ((111)In), gallium ((67/68)Ga), gadolinium (Gd) and so on, complexes of DOTA-conjugated bisphosphonate with various metals could be useful as agents for palliation of metastatic bone pain, bone scintigraphy and magnetic resonance imaging. PMID- 19217525 TI - Tumor targeting and SPECT imaging properties of an (111)In-labeled galectin-3 binding peptide in prostate carcinoma. AB - INTRODUCTION: Galectin-3 (gal-3) is a carbohydrate binding protein that has been implicated in cell adhesion, tumor invasion and metastasis. The objective of this study was to evaluate the tumor targeting and imaging properties of a gal-3 binding peptide selected by phage display in a mouse model of metastatic human prostate carcinoma expressing gal-3. METHODS: A gal-3 binding peptide, ANTPCGPYTHDCPVKR, was synthesized with a Gly-Ser-Gly (GSG) spacer and 1,4,7,10 tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid (DOTA) and then radiolabeled with (111)In. The in vitro cell binding properties of (111)In-DOTA-(GSG) ANTPCGPYTHDCPVKR were determined in metastatic human PC3-M prostate carcinoma cells. The pharmacokinetics and single-photon emission computed tomographic (SPECT/CT) imaging with the radiolabeled peptide were evaluated in SCID mice bearing human PC3-M prostate carcinoma tumor xenografts. RESULTS: The radiolabeled peptide bound with a 50% inhibitory concentration of 191+/-10.2 nM to cultured PC3-M prostate carcinoma cells. In vivo tumor uptake and retention coupled with fast whole-body clearance of the peptide were demonstrated in PC3-M tumor-bearing SCID mice. The tumor uptake rates of the radiolabeled peptide were 1.27+/-0.10%ID/g at 30 min, 0.82+/-0.15%ID/g at 1 h and 0.57+/-0.09%ID/g at 2 h. MicroSPECT/CT studies revealed good tumor uptake of (111)In-DOTA-(GSG) ANTPCGPYTHDCPVKR 2 h postinjection, while uptake in normal organs was low, with the exception of the kidneys. CONCLUSIONS: In vitro cell binding along with tumor uptake of (111)In-DOTA-(GSG)-ANTPCGPYTHDCPVKR in PC3-M human prostate carcinoma tumor-bearing SCID mice suggests the potential of this peptide as a radiopharmaceutical for imaging of gal-3-expressing prostate tumors. PMID- 19217526 TI - Fluorine-18 labeling and biodistribution studies on peroxisome proliferator activated receptor-gamma ligands: potential positron emission tomography imaging agents. AB - INTRODUCTION: Peroxisome proliferator-activated receptor-gamma (PPARgamma) is an important regulator of lipid metabolism; it controls the differentiation of preadipocytes and is also found at high levels in small metastatic tumors. In this report, we describe the radiochemical synthesis and evaluation of two (18)F labeled analogs of the potent and selective PPARgamma agonist farglitazar. MATERIALS AND METHODS: The isomeric aromatic fluorine-substituted target compounds [(2S)-(2-benzoylphenylamino)-3-(4-(2-[2-(4-[(18)F]fluorophenyl)-5 methyloxazol-4-yl]ethoxy)-phenyl)propionic acid ([(18)F]-1) and (2S)-[2-(4 fluorobenzoyl)phenylamino]-3-(4-[2-(5-methyl-2-phenyloxazol-4-yl)ethoxy] phenyl)propionic acid ([(18)F]-2)] were prepared in fluorine-18-labeled form, respectively, by radiofluorination of an iodonium salt precursor or by Ullmann type condensation with 2-iodo-4'-[(18)F]fluorobenzophenone after nucleophilic aromatic substitution with [(18)F]fluoride ion. Each compound was obtained in high specific activity and good radiochemical yield. RESULTS AND DISCUSSION: (18)F-1 and (18)F-2 have high and selective PPARgamma binding affinities comparable to that of the parent molecule farglitazar, and they were found to have good metabolic stability. Tissue biodistribution studies of (18)F-1 and (18)F-2 were conducted, but PPARgamma-mediated uptake of both agents was minimal. CONCLUSION: This study completes our first look at an important class of PPARgamma ligands as potential positron emission tomography (PET) imaging agents for breast cancer and vascular disease. Although (18)F-1 and (18)F-2 have high affinities for PPARgamma and good metabolic stability, their poor target-tissue distribution properties, which likely reflect their high lipophilicity combined with the low titer of PPARgamma in target tissues, indicate that they have limited potential as PPARgamma PET imaging agents. PMID- 19217527 TI - Preparation and evaluation of ethyl [(18)F]fluoroacetate as a proradiotracer of [(18)F]fluoroacetate for the measurement of glial metabolism by PET. AB - INTRODUCTION: Changes in glial metabolism in brain ischemia, Alzheimer's disease, depression, schizophrenia, epilepsy and manganese neurotoxicity have been reported in recent studies. Therefore, it is very important to measure glial metabolism in vivo for the elucidation and diagnosis of these diseases. Radiolabeled acetate is a good candidate for this purpose, but acetate has little uptake in the brain due to its low lipophilicity. We have designed a new proradiotracer, ethyl [(18)F]fluoroacetate ([(18)F]EFA), which is [(18)F]fluoroacetate ([(18)F]FA) esterified with ethanol, to increase the lipophilicity of fluoroacetate (FA), allowing the measurement of glial metabolism. METHODS: The synthesis of [(18)F]EFA was achieved using ethyl O-mesyl glycolate as precursor. The blood-brain barrier permeability of ethyl [1 (14)C]fluoroacetate ([(14)C]EFA) was estimated by a brain uptake index (BUI) method. Hydrolysis of [(14)C]EFA in the brain was calculated by the fraction of radioactivity in lipophilic and water fractions of homogenized brain. Using the plasma of five animal species, the stability of [(14)C]EFA was measured. Biodistribution studies of [(18)F]EFA in ddY mice were carried out and compared with [(18)F]FA. Positron emission tomography (PET) scanning using common marmosets was performed for 90 min postadministration. At 60 min postinjection of [(18)F]EFA, metabolite studies were performed. Organs were dissected from the marmosets, and extracted metabolites were analyzed with a thin-layer chromatography method. RESULTS: The synthesis of [(18)F]EFA was accomplished in a short time (29 min) and with a reproducible radiochemical yield of 28.6+/-3.6% (decay corrected) and a high radiochemical purity of more than 95%. In the brain permeability study, the BUI of [(14)C]EFA was 3.8 times higher than that of sodium [1-(14)C]fluoroacetate. [(14)C]EFA was hydrolyzed rapidly in rat brains. In stability studies using the plasma of five animal species, [(14)C]EFA was stable only in primate plasma. Biodistribution studies in mice showed that the uptake of [(18)F]EFA in selected organs was higher than that of [(18)F]FA. From nonprimate PET studies, [(18)F]EFA was initially taken into the brain after injection. Metabolites related to the tricarboxylic acid (TCA) cycle were detected in common marmoset brain. CONCLUSION: [(18)F]EFA rapidly enters the brain and is then converted into TCA cycle metabolites in the brains of common marmosets. [(18)F]EFA shows promise as a proradiotracer for the measurement of glial metabolism. PMID- 19217528 TI - 3'-Deoxy-3'-[(18)F]fluorothymidine (FLT) uptake in breast cancer cells as a measure of proliferation after doxorubicin and docetaxel treatment. AB - INTRODUCTION: The nucleoside analogue [(18)F]fluorothymidine (FLT) has been designed as a marker of cell proliferation that can be imaged in vivo by positron emission tomography. Clinical pilot studies have demonstrated decreasing FLT uptake following antiproliferative chemotherapy of breast cancer. However, the significance of posttreatment FLT uptake has not been evaluated at the cell level. The aim of this study was to investigate whether FLT uptake detects proliferation inhibition induced by docetaxel or doxorubicin treatment in an in vitro breast cancer model. METHODS: Breast cancer cells (MCF-7) were treated with docetaxel or doxorubicin for 24 h at drug doses inducing 25-99% inhibition of clonogenic survival (IC(25) to IC(99)). Cellular FLT uptake was estimated at 4 h and at 1, 3 and 5 days interval from chemotherapy. [(3)H]Thymidine incorporation and S-phase fraction were measured for comparison. Analysis of variance and the Bland-Altman difference plot were employed for statistical analysis. RESULTS: After treatment, FLT uptake was declined in dependence of the proliferation inhibition mediated by both chemotherapeutic agents (all P<.0001). The decrease of FLT was greater after doxorubicin treatment than after the corresponding docetaxel dose. With doxorubicin (IC(99)), FLT accumulation was reduced by 70% as early as 4 h after treatment. FLT uptake was closely correlated to [(3)H]thymidine incorporation and S-phase fraction (r=.84 to .93). CONCLUSIONS: Right after docetaxel or doxorubicin treatment, FLT uptake corresponds to the reduction of tumor cell proliferation induced. [(18)F]FLT appears promising for monitoring chemosensitivity in breast cancer. PMID- 19217529 TI - In vitro and in vivo analysis of [(64)Cu-NO2A-8-Aoc-BBN(7-14)NH(2)]: a site directed radiopharmaceutical for positron-emission tomography imaging of T-47D human breast cancer tumors. AB - INTRODUCTION: Human breast cancer, from which the T-47D cell line was derived, is known to overexpress the gastrin-releasing peptide receptor (GRPR) in some cases. Bombesin (BBN), an agonist for the GRPR, has been appended with a radionuclide capable of positron-emission tomography (PET) imaging and therapy. (64)Cu-NO2A-8 Aoc-BBN(7-14)NH(2) (NO2A=1,4,7-triazacyclononane-1,4-diacetate) has produced high quality microPET images of GRPR-positive breast cancer xenografted tumors in mice. METHODS: The imaging probe was synthesized by solid-phase peptide synthesis followed by manual conjugation of the 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA) bifunctional chelator and radiolabeling in aqueous solution. The radiolabeled conjugate was subjected to in vitro and in vivo studies to determine its specificity for the GRPR and its pharmacokinetic profile. A T-47D tumor bearing mouse was imaged with microPET/CT and microMRI imaging. RESULTS: The (64)Cu-NO2A-8-Aoc-BBN(7-14)NH(2) targeting vector was determined to specifically localize in GRPR-positive tissue. Accumulation was observed in the tumor in sufficient quantities to allow for identification of tumors in microPET imaging procedures. For example, uptake and retention in T-47D xenografts at 1, 4 and 24 h were determined to be 2.27+/-0.08, 1.35+/-0.14 and 0.28+/-0.07 % ID/g, respectively. CONCLUSIONS: The (64)Cu-NO2A-8-Aoc-BBN(7-14)NH(2) produced high quality microPET images. The pharmacokinetic profile justifies investigation of this bioconjugate as a potentially useful diagnostic/therapeutic agent. Additionally, the bioconjugate would serve as a good starting point for modification and optimization of similar agents to maximize tumor uptake and minimize nontarget accumulation. PMID- 19217530 TI - Measurement of glucose metabolism in rat spinal cord slices with dynamic positron autoradiography. AB - We attempted to measure the regional metabolic rate of glucose (MRglc) in sliced spinal cords in vitro. The thoracic spinal cord of a mature Wister rat was cut into 400-mum slices in oxygenated and cooled (1-4 degrees C) Krebs-Ringer solution. After at least 60 min of preincubation, the spinal cord slices were transferred into double polystyrene chambers and incubated in Krebs-Ringer solution at 36 degrees C, bubbled with 5% O(2)/5% CO(2) gas. To measure MRglc, we used the dynamic positron autoradiography technique (dPAT) with F-18-2-fluoro-2 deoxy-d-glucose ([(18)F]FDG) and the net influx constant of [(18)F]FDG as an index. Uptake curves of [(18)F]FDG were well fitted by straight lines for more than 7 h after the slicing of the spinal cord (linear regression coefficient, r=0.99), indicating a constant uptake of glucose by the spinal cord tissue. The slope (K), which denotes MRglc, is affected by tetrodotoxin, and high K(+) (50 mM) or Ca(2+)-free, high Mg(2+) solution. After 10 min of hypoxia, the K value following reoxygenation was similar to the unloaded control value, but after 45 min of hypoxia, the K value was markedly lower than the unloaded control value, and after >90 min of reoxygenation it was nearly 0. Our results indicate that the living spinal cord slices used retained an activity-dependent metabolism to some extent. This technique may provide a new approach for measuring MRglc in sliced living spinal cord tissue in vitro and for quantifying the dynamic changes in MRglc in response to various interventions such as hypoxia. PMID- 19217531 TI - Expressions of glucose transporter Types 1 and 3 and hexokinase-II in diffuse large B-cell lymphoma and other B-cell non-Hodgkin's lymphomas. AB - INTRODUCTION: Diffuse large B-cell lymphoma (DLBCL) has been reported to show higher uptake of 2-deoxy-2-F18-fluoro-d-glucose (FDG) by positron emission tomography than other B-cell non-Hodgkin's lymphomas (non-DLBCL). The authors addressed the mechanism of FDG uptake in DLBCL by immunostaining for glucose transporter Types 1 (Glut-1) and 3 (Glut-3) and hexokinase-II (HK-II) in excised lymphoma tissues. METHODS: Sixteen B-cell non-Hodgkin's lymphoma patients (11 DLBCL and 5 non-DLBCL patients) were included in the study because the lymphoma tissues obtained by excision were eligible for immunostaining. The expressions of Glut-1, Glut-3 and HK-II were assessed regarding the percentages of positively stained lymphoma cells (%expression), the staining intensities (none=0, weak=1, moderate=2 and strong=3) and the staining patterns (membranous or cytoplasmic) and compared between DLBCL and non-DLBCL. RESULTS: Glut-1 was not expressed at all in DLBCL or non-DLBCL, and their Glut-3 expressions were not significantly different (P>.05) with respect to %expression (mean+/-S.E.M., 73.6+/-7.3% vs. 72.0+/-3.7%), staining intensity (2.5+/-0.2 vs. 2.6+/-0.2) and staining pattern (membranous pattern dominant; 54.5% vs. 60.0%). However, DLBCL expressed more HK II than non-DLBCL, i.e., %expression (45.2+/-11.5% vs. 17.0+/-15.8%, P=.0275) and staining intensity (2.3+/-0.2 vs. 0.6+/-0.4, P=.0032). HK-II showed a cytoplasmic location in DLBCL and non-DLBCL. CONCLUSIONS: HK-II and Glut-3 contribute significantly to FDG uptake in DLBCL. DLBCL may have higher FDG uptake because it expresses more HK-II, whereas Glut-1 appears to play no role in FDG uptake in B cell non-Hodgkin's lymphoma. PMID- 19217532 TI - Clinical evaluation of antimicrobial peptide [(99m)Tc/Tricine/HYNIC(0)]ubiquicidin 29-41 as a human-specific infection imaging agent. AB - INTRODUCTION: Ubiquicidin (UBI) 29-41 is a cationic synthetic antimicrobial peptide fragment that binds preferentially with the anionic microbial cell membrane at the site of infection. This study was conducted to evaluate the potentiality of [(99m)Tc/Tricine/HYNIC(0)]UBI 29-41 prepared from lyophilized kits as an infection imaging agent in humans. METHODS: Seven patients (5 males and 2 females; mean age=55 years; age range=35-75 years) with suspected bone or soft-tissue infections participated in this study. [(99m)Tc/Tricine/HYNIC]UBI 29 41, corresponding to activity in the range 555-740 MBq added to 40 mug of peptide obtained from instant freeze-dried kit formulations with radiochemical purities >95%, was injected intravenously. A 45-min dynamic study was followed by spot views of the suspected region of infection (target) and a corresponding normal area (nontarget). Whole-body anterior and posterior images were also acquired at 30, 60 and 120 min after injection. True- or false-positive or true- or false negative images were interpreted upon bacterial culture, radiography, clinical tests and bone scanning. RESULTS: The biodistribution of [(99m)Tc/Tricine/HYNIC]UBI 29-41 in patients showed rapid accumulation of activity in the kidneys in the first 30 min after injection that gradually declined and accumulated in the urinary bladder. There were positive findings in five studies and negative findings in two. Findings were subsequently confirmed to be true positive or negative. Images showed minimal accumulation in nontarget tissues, with an average target/nontarget ratio of 2.10+/-0.33 in positive lesions at 30 min. CONCLUSION: Given its favorable clinical characteristics, [(99m)Tc/Tricine/HYNIC]UBI 29-41 shows promise as a tracer for infection imaging that allows early diagnosis (30 min) of infection. PMID- 19217533 TI - Comparative study of cellular kinetics of reporter probe [(131)I]FIAU in neonatal cardiac myocytes after transfer of HSV1-tk reporter gene with two vectors. AB - AIM: Reporter gene imaging is a promising approach for noninvasive monitoring of cardiac gene therapy. In this study, HSV1-tk (herpes simplex virus type 1 thymidine kinase) and FIAU (2'-fluoro-2'-deoxy-1-beta-d-arabinofuranosyl-5 iodouracil) were used as the reporter gene and probe, respectively. Cellular uptakes of radiolabeled FIAU of neonatal rat cardiac myocytes transferred with HSV1-tk were compared between two vectors, adenovirus and liposome. The aims of this study were to choose the better vector and to provide a theoretical basis for good nuclide images. METHODS: Neonatal cardiac myocytes were obtained from rat heart by single collagenase digestion. HSV1-tk inserted into adenovirus vector (recombinant adenovirus type 5, Ad5-tk) and plasmid (pDC316-tk) coated with Lipofectamine 2000 (pDC316-tk/lipoplex) were developed; thus, HSV1-tk could be transferred into neonatal cardiac myocytes. FAU (2'-fluoro-2'-deoxy-1-beta-d arabinofuranosyluracil) was labeled with (131)I, and the product was assessed after purification with reversed-phase Sep-Pak C-18 column. The uptake rates of [(131)I]FIAU in the transferred cardiac myocytes at different times (0.5, 1, 2, 3, 4 and 5 h) were detected. Furthermore, mRNA expression and protein expression of HSV1-tk were detected by semiquantitative reverse-transcriptase polymerase chain reaction and immunocytochemistry. RESULTS: FAU could be labeled with (131)I, and the labeling efficiency and radiochemical purity rates were 53.82+/ 2.05% and 94.85+/-1.76%, respectively. Time-dependent increase of the accumulation of [(131)I]FIAU was observed in both the Ad5-tk group and the pDC316/lipoplex group, and the highest uptake rate occurred at 5 h, with peak values of 12.55+/-0.37% and 2.09+/-0.34%, respectively. Greater uptakes of [(131)I]FIAU in Ad5-tk-infected cells compared with pDC316/lipoplex-transfected ones occurred at all the time points (t=12.978-38.253, P<.01). The exogenous gene expression by polymerase chain reaction in adenovirus vector-infected cardiac myocytes was significantly higher than that in pDC316-tk/lipoplex-transducted ones (semiquantitative analysis, 3.11+/-0.14 versus 1.60+/-0.05, P<.01). Immunocytochemistry showed that the transferred cardiac myocytes successfully expressed the target protein, and the positive rates were 81.70+/-0.40% in Ad5-tk and 22.06+/-0.32% in liposome (P<.01). CONCLUSIONS: Both adenovirus and liposome could transfer reporter gene into cardiac myocytes successfully, and the expressed exogenous protein could form functional enzymes efficiently. However, the adenovirus vector acted more efficiently than did liposome, with a higher uptake rate of the reporter probe. Thus, adenovirus is competent for gene transfer in cardiac reporter gene imaging. PMID- 19217534 TI - Synthesis and PET studies of [(11)C-cyano]letrozole (Femara), an aromatase inhibitor drug. AB - INTRODUCTION: Aromatase, a member of the cytochrome P450 family, converts androgens such as androstenedione and testosterone into estrone and estradiol, respectively. Letrozole (1-[bis-(4-cyanophenyl)methyl]-1H-1,2,4-triazole; Femara) is a high-affinity aromatase inhibitor (K(i)=11.5 nM) that has Food and Drug Administration approval for breast cancer treatment. Here we report the synthesis of carbon-11-labeled letrozole and its assessment as a radiotracer for brain aromatase in the baboon. METHODS: Letrozole and its precursor (4-[(4-bromophenyl) 1H-1,2,4-triazol-1-ylmethyl]benzonitrile) were prepared in a two-step synthesis from 4-cyanobenzyl bromide and 4-bromobenzyl bromide, respectively. The [(11)C]cyano group was introduced via tetrakis(triphenylphosphine)palladium(0) catalyzed coupling of [(11)C]cyanide with the bromo precursor. Positron emission tomography (PET) studies in the baboon brain were carried out to assess regional distribution and kinetics, reproducibility of repeated measures and saturability. Log D, the free fraction of letrozole in plasma and the [(11)C-cyano]letrozole fraction in arterial plasma were also measured. RESULTS: [(11)C-cyano]Letrozole was synthesized in 60 min with a radiochemical yield of 79-80%, with a radiochemical purity greater than 98% and a specific activity of 4.16+/-2.21 Ci/mumol at the end of bombardment (n=4). PET studies in the baboon revealed initial rapid and high uptake and initial rapid clearance, followed by slow clearance of carbon-11 from the brain, with no difference between brain regions. Brain kinetics was not affected by coinjection of unlabeled letrozole (0.1 mg/kg). The free fraction of letrozole in plasma was 48.9%, and log D was 1.84. CONCLUSION: [(11)C-cyano]Letrozole is readily synthesized via a palladium catalyzed coupling reaction with [(11)C]cyanide. Although it is unsuitable as a PET radiotracer for brain aromatase, as revealed by the absence of regional specificity and saturability in brain regions such as amygdala, which are known to contain aromatase, it may be useful in measuring letrozole distribution and pharmacokinetics in the brain and peripheral organs. PMID- 19217537 TI - Leadership is not just a title or position; it is also an attitude. PMID- 19217535 TI - Development and validation of 2-deoxy-2-chloro-d-glucose impurity analysis in [(18)F]FDG by three potential-time waveforms of high-performance liquid chromatography/pulsed amperometric detection. AB - A suitable three potential-time waveforms for the electrochemical detection of 2 deoxy-2-chloro-d-glucose (ClDG) by gold working electrode and palladium reference electrode have been developed and method validation was performed on Waters 2796 Bioalliance HPLC system coupled with pulsed amperometric detection (HPLC/PAD). FDG and ClDG could be completely separated by 50 mM NaOH mobile phase at a flow rate of 1.0 ml/min; 30 degrees C analytical column temperature; and E1 of 200 mV, T1 of 900 mS; E2 of -770 mV, T2 of 10 mS; E3 of 1400 mV, T3 of 10 mS; acquisition delay (AD) of 300 mS. The validation results were shown as follows: (1) in specificity study, mannose, FDG and ClDG could be completely separated and the retention times of these were 6.2, 11.1 and 13.5 min, respectively, with a total run time of 20 min; (2) the intraday repeatable precision expressed with the CV% in six successive analysis was 0.52% (for FDG) and 0.83% (for ClDG); (3) the interday variability precision expressed with the CV% value of the repeatable precision of 3 days was 0.99%, 0.52% and 0.58% for FDG and 0.71%, 0.83% and 1.24% for ClDG; both the CV% of intraday and interday reproducibilities of FDG and ClDG were better than 1.5%; (4) the accuracy and recovery of FDG and ClDG expressed with the percentage of mean value of three successive analysis were 99.75% (for FDG) and 100.68% (for ClDG) which were all greater than 95%; (5) under optimum conditions, the limit of detection of FDG and ClDG was 0.41 and 0.68 microg/ml, and the limit of quantization of FDG and ClDG was 1.24 and 2.04 microg/ml; (6) the correlation coefficient (r) value of linearity is over 0.999 by 5-50 microg/ml ranges of both compounds, respectively; (7) no interference peak effects by composition of mobile phase or increasing/decreasing flow rate or change of temperature was observed. PMID- 19217538 TI - Changes in pain-free walking based on time in accommodating pain-free exercise therapy for peripheral arterial disease. AB - Symptoms of intermittent claudication (IC) can be relieved by lifestyle modification, medications, and walking exercises. The intensity of the walking exercise is still debatable. The goal of this study was to determine the effects of accommodating pain-free walking exercise therapy program length on pain-free walking. A descriptive, longitudinal study with repeated measures of exercise capacity was conducted. An IC questionnaire was administered to assess IC signs, symptoms, and lifestyle. Walking was performed on a treadmill for 30 to 50 minutes below the participant's individualized walking pain threshold. The study included patients diagnosed with IC due to peripheral arterial disease. All participants were randomly assigned to three groups. Group A (n = 28) participated in the walking program for 2-9 weeks, group B (n = 30) for 10-14 weeks, and group C (n = 26) for 15-94 weeks. The main outcome measure of the study was to determine changes in exercise capacity: walking distance (miles), walking duration (minutes), and walking speed (mph). Group A increased the amount of distance, duration, and speed walked from pretest to posttest by 80% (P < .001), 27% (P < .001), and 37% (P < .001), respectively. Group B increased the amount of distance, duration, and speed walked from pretest to posttest by 122% (P < .001), 56% (P < .001), and 43% (P < .001), respectively. Group C increased the amount of distance, duration, and speed walked from pretest to posttest by 26% (P = .002), 22% (P = .002), and 5% (P = .541) respectively. We reached the conclusion that a walking program of 10-14 weeks is optimal for achieving the best improvements in walking distance, duration, and speed. PMID- 19217539 TI - Continuous improvement process for a high-risk population: catheter-directed thrombolytic infusions. AB - Catheter directed infusion of thrombolytics, though beneficial in restoring blood flow to an ischemic limb, remains a high-risk procedure requiring vigilant, coordinated care to prevent complications. A multidisciplinary team that included staff from all care settings and disciplines was formed. This team used a continuous improvement process to identify system problems, analyze work processes and develop tools to standardize and guide care. Because catheter directed peripheral thrombolytic infusion is a low volume, high-risk procedure, data were collected concurrently over 4 years on all patients. Concurrent data collection identified problems as they occurred, allowing problem analysis and resolution to begin immediately. The outcomes measured included time to initiation of thrombolytic (goal was 30 minutes or less) and the incidence of bleeding complications. Data from 61 cases from November 2003 through November 2007 were analyzed. Although the 30-minute goal for the initiation of the thrombolytic infusion was met for some time periods, it was difficult to sustain and required continued communication and collaboration. The incidence of bleeding complications were within an expected range. Concurrent data collection with real time problem solving in a team setting enhanced the delivery of safe, effective care to a high-risk, low-volume population. PMID- 19217540 TI - Foot venous system arterialization for salvage of nonreconstructable acute ischemic limb: a case report. AB - Patients 73 years old man, was admitted to the Hospital with acute ischaemia of the left lower limb, pain started 4 days ago. Ultrasound investigation and lower limbs angiography were performed. Giant infrapopliteal aneurysm and thrombosis due to massive embolism of the crural and pedal arteries' was revealed. In order to keep the limb, urgent surgical treatment--popliteal aneurysm ligation and foot venous system arterialisation was performed. Urgent venous arterialisation may be considered as viable alternative in patients with limb acute ischaemia, when arterial reconstruction is impossible to perform. In our case improvement of the foot perfusion during 8 days increases the interest to this technique. PMID- 19217541 TI - Review of an article: Canonico M, Plu-Bureau G, Lowe G, Scarabin P. (2008). Hormone replacement therapy and risk of venous thromboembolism in postmenopausal women: systematic review and meta-analysis. British Medical Journal 10(336):1227 1231. PMID- 19217542 TI - Practical tips for starting a journal club. PMID- 19217543 TI - Identification of mutation sites on Delta12 desaturase genes from Mortierella alpina 1S-4 mutants. AB - The mutation sites on the Delta12 desaturase gene in Mortierella alpina Delta12 desaturase-defective mutants SR88, TM912, and Mut48 accumulating Mead acid were identified. Each mutation resulted in an amino acid replacement (H116Y and P166L) in the Delta12 desaturase gene from SR88 and Mut48, respectively. PMID- 19217544 TI - Enhancement of over expression and chaperone assisted yield of folded recombinant aconitase in Escherichia coli in bioreactor cultures. AB - A major portion of the over expressed yeast mitochondrial aconitase, a large 82 kDa monomeric TCA cycle enzyme, in Escherichia coli led to the formation of inclusion bodies. Bacterial chaperonin GroEL mediated the correct folding of aconitase with the assistance of its co-chaperonin GroES in an ATP dependent manner. Till date the chaperonin assisted folding of aconitase was limited to the shake flask studies with relatively low yields of folded aconitase. No attempt had yet been made to enhance the yield of chaperone mediated folding of aconitase using a bioreactor. The current report deals with the effect of co-expression of GroEL/GroES in the production of soluble, biologically active recombinant aconitase in E. coli by cultivation in a bioreactor at different temperatures under optimized conditions. It revealed that the yield of functional aconitase was enhanced, either in presence of co-expressed GroEL/ES or at low temperature cultivation. However, the outcome from the chaperone assisted folding of aconitase was more pronounced at lower temperature. A 3-fold enhancement in the yield of functional aconitase from the bioreactor based chaperone assisted folding was obtained as compared to the shake flask study. Hence, the present study provides optimized conditions for increasing the yield of functional aconitase by batch cultivation in a bioreactor. PMID- 19217545 TI - Production and characterization of alcohol oxidase from Penicillium purpurescens AIU 063. AB - A filamentous fungus belonging to Penicillium purpurescens was isolated as a strain grown well in methanol medium. The strain produced an alcohol oxidase (AOD), which oxidizes short-chain primary alcohols and ethylene glycol, by incubation with a medium containing not only methanol, but also ethanol, ethylene glycol, glycerol or glucose. This AOD belonged to the same group as AOD from methylotrophic yeast, but some properties were different from AODs from methylotrophic yeast and other fungi. Thus, this enzyme consisted of four identical subunits of 66 kDa, while AODs from methylotrophic yeasts were of eight subunits. The enzyme activity was enhanced to two-fold by incubation at pH 6.0 and 40 degrees C for 60 min, whereas the K(m) values for ethanol and ethylene glycol did not change. PMID- 19217546 TI - Purification and characterization of two DyP isozymes from Thanatephorus cucumeris Dec 1 specifically expressed in an air-membrane surface bioreactor. AB - DyP isozymes (DyP2 and DyP3) from the culture fluid of the fungus Thanatephorus cucumeris Dec 1 by air-membrane surface bioreactor were purified and characterized. The characteristics of DyP2 were almost the same as those of a recombinant DyP reported previously, but different from DyP3. PMID- 19217547 TI - Purification and characterization of a (S)-1-phenyl-1, 3-propanediol-producing enzyme from Williopsis saturnus var. mrakii AJ-5620. PMID- 19217548 TI - Streptococcus thermophilus produces exopolysaccharides including hyaluronic acid. AB - Hyaluronic acid (HA) is an important material for medical, cosmetic and food applications. HA is obtained commercially from rooster combs and the fermentation of streptococci. However, the safety problems such as hyaluronidase or exotoxin contamination remain controversial. To reduce the risk of hyaluronidase or exotoxin contamination, we attempted to isolate strains of Streptococcus thermophilus with high productivity of useful exopolysaccharides (EPSs) including HA from traditional dairy food products. Forty-six S. thermophilus strains were isolated from dairy food products, and examined of their HA production using a HA binding protein method. According to the results, six S. thermophilus strains produced EPSs including HA. S. thermophilus YIT 2084 had a markedly high HA productivity (approximately 8 mg/l). We focused on the high-molecular-mass fraction of EPS (2000 kDa) from S. thermophilus YIT 2084. By using high performance liquid chromatography, it was found that a high-molecular-mass fraction of EPS included N-acetylglucosamine (54.4%) and glucuronic acid (45.6%), which are components of HA. Furthermore, 13C-nuclear magnetic resonance spectroscopy showed that the spectra of the high-molecular-mass fraction corresponded well to those of commercial HA. Here, we described for the first time that S. thermophilus, which is a generally recognized safe bacterium, produces HA. The novel HA-producing bacterium S thermophilus YIT 2084 has great potential for applications in the medical, cosmetic and food fields, although its culture conditions remain to be improved. PMID- 19217549 TI - Characterization of the C-terminal truncated form of amylopullulanase from Lactobacillus plantarum L137. AB - A gene (apuA) encoding amylopullulanase from a starch-hydrolyzing lactic acid bacterium, Lactobacillus plantarum L137, which had been isolated from traditional fermented food made from fish and rice in the Philippines, was found to contain two unique amino acid repeating units in the N- and C-terminal region. The former is a six amino acid sequence (Asp-Ala/Thr-Ala-Asn-Ser-Thr) repeated 39 times, and the latter is a three amino acid sequence (Gln-Pro-Thr) repeated 50 times. To clarify the role of these repeating units, a truncated apuA in the C-terminal region was constructed and expressed in L. plantarum NCL21, which is the ApuA- derivative of strain L137. The recombinant truncated amylopullulanase (ApuADelta), which lacks the 24 kDa of the C-terminal repeat region, was purified and characterized, and compared with wild-type amylopullulanase (ApuA). The enzyme production and specific activity of ApuADelta were higher than those of ApuA. The two enzymes, ApuA and ApuADelta, showed similar pH (4.0-4.5) and temperature (40-45 degrees C) optima. However, the activity of ApuADelta was more stable in the pH and temperature than that of ApuA. The catalytic efficiencies of ApuADelta toward soluble starch, pullulan and amylose were higher than those of ApuA, although their substrate specificities towards saccharides were similar. From these results, we conclude that the C-terminal repeating region of ApuA is negatively involved in the stability of amylopullulanase and binding of substrates. Thus, the truncated amylopullulanase is more useful in processing of amylose and pullulan. PMID- 19217550 TI - Slurry bioreactor modeling using a dissimilatory arsenate-reducing bacterium for remediation of arsenic-contaminated soil. AB - A slurry bioreactor using a dissimilatory arsenate (As(V))-reducing bacterium is proposed for remediation of arsenic-contaminated soils. Bacterial As(V) reduction can cause arsenic extraction from the solid to the liquid phase because arsenite, As(III), is much less adsorptive than As(V). A mathematical model was developed incorporating the reversible sorption process of arsenic as well as bacterial growth and decay via As(V) reduction. A linear isotherm equation expressed the sorption process. The model included Haldane kinetics with high As(V) concentrations and cell inactivation by toxicity due to As(III). Extraction experiments used synthetic contaminated soils (forest soil, Soil SF, 1100 mg kg( 1); paddy soil, Soil SP, 1100 mg kg(-1)) and actual contaminated soils (Soil AH 2200 mg kg(-1) and Soil AL, 220 mg kg(-1)) at 5% w/v slurry concentration. Simulation results matched the observed changes of arsenic concentrations in the liquid phase. The respective extraction efficiencies of arsenic were 63%, 41%, 20%, and 55% for SF, SP, AH, and AL soils. Sensitivity analyses showed that the rate-limiting step was the desorption rate of As(V) from the solid to the liquid phase, rather than the As(V)-reducing rate. The proposed model provides a useful framework for understanding and predicting the extraction of arsenic from soil. PMID- 19217551 TI - Process optimization of biological hydrogen production from molasses by a newly isolated Clostridium butyricum W5. AB - This work sought to optimize fermentation parameters in a batch process for hydrogen production from molasses by a newly isolated Clostridium butyricum W5. Hydrogen yield and production rate, bacterial biomass and volatile fatty acids, including acetic, lactic and butyric acids, were measured. Key fermentation operation parameters, including concentration of carbon and nitrogen sources, growth temperature and pH, and inoculum size were investigated. The best results in terms of hydrogen yield and productivity were obtained under the conditions of 100 g/L molasses, 1.2 g/L NH4NO3, 39 degrees C at pH 6.5 with initial cell concentration of 9x10(4) cell/ml. Maximum hydrogen yield was 1.85 mol hydrogen/mol hexose, corresponding to a hydrogen production rate of 17.38 mmol/h/L. Experimental data showed that the acetic/butyric acid ratio remained relatively stable with an increase in molasses concentration, while the unfavoured product, lactic acid, portion increased. No solvent (ethanol, butanol and acetone) was detected during the fermentation. Propionic acid was measured at a very low level in the hydrogen fermentation. Statistical analysis showed that hydrogen yield increased exponentially with the increase in cell growth, and that there was no correlation between the hydrogen yield and ratio of acetic acid to butyric acid. PMID- 19217552 TI - Rhodococcus aetherivorans IAR1, a new bacterial strain synthesizing poly(3 hydroxybutyrate-co-3-hydroxyvalerate) from toluene. AB - Toluene is the volatile organic compound (VOC) that is most abundantly emitted to the environment. Here, we report a new bacterial strain, IAR1, which synthesizes polyhydroxyalkanoate (PHA) from toluene as the sole carbon source. Using physiological tests and phylogenetic analysis, IAR1 was identified as Rhodococcus aetherivorans. Degradation of toluene by resting IAR1 cells followed Michaelis Menten kinetics. The determined kinetic parameters showed that the degradation ability of IAR1 is slightly lower than that of Acinetobacter sp. Tol 5, which is highly effective for removing toluene from off-gas. PHA synthesized by IAR1 was identified as poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) by 1H NMR and gas chromatography. During fed-batch cultivation of resting IAR1 cells, the principal monomer unit shifted from 3-hydroxybutyrate to 3-hydroxyvalerate. PHBV shows flexibility and toughness comparable to conventional thermoplastics, but its production usually requires supplying expensive precursors as a secondary carbon source. Therefore, employment of R. aetherivorans IAR1 in the production of PHBV from toluene as the sole carbon source is an attractive means for production of the practical copolymer. In addition, the utilization of waste toluene as a raw material for the polymer production will achieve further cost reduction alongside the effective utilization of waste material. PMID- 19217553 TI - Removal of Mn2+ from water by "aged" biofilter media: the role of catalytic oxides layers. AB - The present work was aimed at evaluating the surface coatings characteristics and autocatalytic manganese oxidation potentials of two groups of "aged" biofilter media. This refers to the anthracite filter media of a biological water treatment plant on which metal oxides and a biofilm have deposited on the surface of the filter media over long time of filtration. Duplicate samples of anthracite filter media were collected from each of the six filter wells in the plant and classified into two groups, based on their duration of operation, as 3-years filter media and 15-years filter media. Batch experiments showed that the 15 years filter media exhibited very high manganese sorption capacity and were less dependent on the microbial activity than the 3-years filter media. Results of the surface coatings analyses indicated that the biofilter materials is predominantly composed of variable layers of manganese and iron oxides, with microbial biomass contributing only about 3.5 and 1.4% of the dry weight of the surface coatings on the 3- and 15-years filter media respectively. Investigations onto the Mn2+ sorption by the lyophilized biofilter media showed that, the sorption kinetics on the catalytic oxides layers followed the pseudo-second-order kinetics model, thus suggesting chemisorption as the dominant mechanism of Mn2+ removal. This implied that manganese removal by these biofilters is mainly by adsorption of Mn2+ onto the iron and manganese (catalytic) oxides layers and autocatalytic oxidation. The present study has clearly linked Mn2+ oxidation to the catalytic oxides layers on the aged biofilter media. PMID- 19217554 TI - Enzymatic hydrolysis of cuttlefish (Sepia officinalis) and sardine (Sardina pilchardus) viscera using commercial proteases: effects on lipid distribution and amino acid composition. AB - Total lipid and phospholipid recovery as well as amino acid quality and composition from cuttlefish (Sepia officinalis) and sardine (Sardina pilchardus) were compared. Enzymatic hydrolyses were performed using the three proteases Protamex, Alcalase, and Flavourzyme by the pH-stat method (24 h, pH 8, 50 degrees C). Three fractions were generated: an insoluble sludge, a soluble aqueous phase, and an oily phase. For each fraction, lipids, phospholipids, and proteins were quantified. Quantitative and qualitative analyses of the raw material and hydrolysates were performed. The degree of hydrolysis (DH) for cuttlefish viscera was 3.2% using Protamex, 6.8% using Flavourzyme, and 7% using Alcalase. DH for sardine viscera was 1.9% (using Flavourzyme), 3.1% (using Protamex) and 3.3% (using Alcalase). Dry matter yields of all hydrolysis reactions increased in the aqueous phases. Protein recovery following hydrolysis ranged from 57.2% to 64.3% for cuttlefish and 57.4% to 61.2% for sardine. Tissue disruption following protease treatment increased lipid extractability, leading to higher total lipid content after hydrolysis. At least 80% of the lipids quantified in the raw material were distributed in the liquid phases for both substrates. The hydrolysed lipids were richer in phospholipids than in the lipids extracted by classical chemical extraction, especially after Flavourzyme hydrolysis for cuttlefish and Alcalase hydrolysis for sardine. The total amino acid content differed according to the substrate and the enzyme used. However, regardless of the raw material or the protease used, hydrolysis increased the level of essential amino acids in the hydrolysates, thereby increasing their potential nutritional value for feed products. PMID- 19217555 TI - Change in glycosylation pattern with extension of endoplasmic reticulum retention signal sequence of mouse antibody produced by suspension-cultured tobacco BY2 cells. AB - The production of antibodies using plants as bioreactors has been realized. Because sugar chain structures on recombinant proteins are a cause of concern, remodeling technology is highly promising. Localizing recombinant proteins in the endoplasmic reticulum (ER) affects the glycosylation pattern in transgenic plants. Previously, a mouse antibody produced by suspension-cultured tobacco BY2 cells has sugar chains with possible glycoepitopes as the predominant structures. In this study, we extended the Lys-Asp-Glu-Leu (KDEL) ER retention signal sequence over the heavy (H) and light (L) chains of the antibody and expressed the altered antibody in tobacco BY2 cells to study the effect of the KDEL sequence on glycosylation. For the antibody with the KDEL-extended H-chains, glycans with beta(1,2)-xylose or alpha(1,3)-fucose residues accounted for 49% of the total glycans. Meanwhile, for the antibody with the KDEL-extended H- and L chains, glycans with xylose or fucose accounted for 38% of the total glycans. Although the addition of an ER retention signal shifted the dominant glycan structures of the KDEL-extended antibody to high-mannose-type structures, some of the antibodies escaped the retrieval system during intracellular traffic and were then modified by xylosylation or fucosylation. PMID- 19217556 TI - Phytochelatins inhibit the metal-induced aggregation of alpha-crystallin. AB - Phytochelatins (PCs) are heavy-metal-binding peptides found in some eukaryotes. This study investigates the use of plant-derived PCs for the inhibition of metal induced protein aggregation. The results of this study show that PCs inhibit zinc induced alpha-crystallin aggregation, and suggest that PCs might be useful as anti-cataract agents. PMID- 19217557 TI - Type II collagen-chondroitin sulfate-hyaluronan scaffold cross-linked by genipin for cartilage tissue engineering. AB - Owing to of the limited repair capacity of articular cartilage, it is essential to develop tissue-engineered cartilage for patients suffering from joint disease. Chondroitin sulfate (CS) and hyaluronan (HA) are the components of the cartilage extracellular matrix (ECM) and are known to influence the proliferation and differentiation of chondrocytes. Scaffolds composed of type-II collagen, CS, and HA may create an environment that can preserve the normal phenotype of cells to promote regeneration of cartilage-like constructs. In this investigation, we prepared and characterized 3-dimensional type-II collagen scaffolds both with and without HA and CS. Porous composite scaffolds fabricated by freeze-drying showed interconnected pores with mean diameters of 140+/-30 microm and porosities of 92 95% after cross-linking with genipin. After a 14-day in vitro culture, morphologically round chondrocytes were found to be uniformly distributed throughout the sponges. Expression of genes of aggrecan, type-II collagen and cartilage oligomeric matrix protein (COMP) was statistically and significantly increased on scaffolds with CS and HA than those without CS and HA. Furthermore, there was a markedly greater accumulation of proteoglycans (PGs) on the scaffolds with CS and HA. PMID- 19217558 TI - Novel bench-top perfusion system improves functional performance of bioengineered heart muscle. AB - Research in the area of cardiac tissue engineering is focused on the development of functional 3-dimensional cardiac muscle tissue in vitro, which includes bioengineered cardiac patches, pumps and ventricles. One of the major challenges in the field of cardiovascular tissue engineering is determining how to support the increased metabolic demands of 3-dimensional tissue constructs, due to the increase in both cellular mass and density compared to monolayer cultures. Traditional culture systems rely on passive diffusion for the delivery of oxygen and soluble factors. However, perfusion systems can provide continuous delivery of cell culture media to 3D tissue constructs, which promotes more active delivery of oxygen, soluble factors, and shear stress, which can be utilized to guide tissue maturation and functional remodeling of bioengineered tissues. We have previously described a perfusion system and demonstrated compatibility over short time periods (approximately hours) with 2-dimensional monolayer cell culture and 3-dimensional tissue constructs. The objectives of our current study were to: introduce CO2 buffering to stabilize media pH in order to achieve long term culture within the system, incorporate sensors capable of recording high media oxygen concentrations, and to increase the culture time of bioengineered heart muscle within the perfusion system in order to increase their functional performance. We showed that exposure of bioengineered heart muscle to perfusion for a period of 24 h increased their functional performance, as measured by cellular viability, total protein, total RNA, spontaneous contractility, twitch force, and specific force. PMID- 19217559 TI - Effect of lactoferrin-embedded collagen membrane on osteogenic differentiation of human osteoblast-like cells. AB - Lactoferrin (LF) has the ability to promote the proliferation and differentiation of osteoblasts, suggesting its potential utility as an osteogenic growth factor in bone tissue engineering. However, this type of application requires improved drug delivery system (DDS) technology at the target site. In this study, we report enhanced calcium deposition and alkaline phosphatase (ALP) activity using the type I collagen membrane during osteogenic differentiation of MG63 human osteoblast-like cells, indicating that type I collagen not only acts as a site for calcification but also promotes the expression of differentiated phenotypes. We also used this membrane as a drug delivery carrier for bovine LF. Approximately 27% of LF embedded on the type I collagen membrane was released within the first hour in cell-free condition. This initial burst release of LF was followed by a slower release from the collagen membrane. Bovine LF embedded in the type I collagen membrane promoted its calcification during osteogenic differentiation of MG63 cells without the loss of LF bioactivity. Taken together, ALP activity and osteocalcin production were enhanced in the MG63 cells plated on the LF-embedded collagen membrane, suggesting that LF incorporated in the collagen membrane promoted bone-like tissue formation by MG63 cells. These observations suggest that the type I collagen membrane is useful as a drug delivery carrier for LF in bone tissue engineering. PMID- 19217560 TI - Morphological regulation and aggregate formation of rabbit chondrocytes on dendrimer-immobilized surfaces with D-glucose display. AB - The present study describes dendrimer-immobilized surfaces with D-glucose display, used to regulate the morphology of rabbit chondrocytes by changing the generation number and density of the dendrimer, and to clarify the relation between cell morphology and chondrogenic expression on the prepared surfaces. When the generation number increased, the frequency of round-shaped cells increased, whereas, cell stretching was appreciably suppressed. Further suppression of stretching was significant on the G4-LD surface, on which the density of the immobilized dendrimer was lowered, and this phenomenon was thought to be responsible for the sparse display of D-glucose on the surface. The time lapse observations revealed that the G4-LD surface caused repeated morphological changes of stretching and contracting, together with fluctuations in cell roundness. By the cytoskeletal staining of F-actin, the immature stress fibers were recognized in both round and stretched cells on the G4-LD surface. It was also found that N-cadherin expression was promoted on this surface, thus supporting the idea that an increase in cell-cell contacts may maintain chondrocytic phenotypes. These results suggest that a G4-LD surface can provide a culture environment to promote cell-cell interactions by N-cadherin-mediated adhesion during cell aggregation, thereby facilitating chondrocytic phenotypes of cells. PMID- 19217561 TI - Enhancement of antibody production from a chicken B cell line DT40 by reducing Pax5 expression. AB - We developed a novel in vitro antibody (Ab) generation system using a hypermutating chicken B cell line (DT40-SW). We suppressed the expression of the Pax5 transcription factor by targeted disruption of the gene to increase Ab production in isolated clones and produce the desired Abs. This single genetic manipulation resulted in a significant enhancement of Ab production without significantly affecting maximum cell density. PMID- 19217562 TI - Identification and validation of suitable reference genes for quantitative expression of xylA and xylE genes in Pseudomonas putida mt-2. AB - Reference genes are used to normalize target genes for relative quantification in gene expression studies. However, different experimental conditions may affect the expression of reference genes, which could lead to erroneous quantitative results. In this study, we performed real-time polymerase chain to investigate the expression of eight reference genes (rpoN, rpoD, dbhA, phaF, 16S rRNA, gst, lexA, and atkA) in Pseudomonas putida mt-2 during degradation of p-xylene. According to their expression stability, geNorm software analysis revealed that rpoN, rpoD, 16S rRNA, and atkA were suitable reference genes with highly stable expression, whereas phaF and dbhA were not suitable due to unstable expression. When normalized either to phaF or dbhA, xylA and xylE expression were significantly different compared to the expression levels normalized with the normalization factor (NF(4)) obtained from the four most stable reference genes (rpoN, -rpoD, -16S rRNA, and -atkA). The use of unstably expressing reference genes resulted in an over- or underestimation of target gene expression, a delay in maximal gene expression, and an increase in gene expression in the absence of inducer. While experimental results indicated that the relative maximum expression of xylA and xylE occurred at different times, unstable reference genes indicated that the maximum expression occurred at the same time. Our study indicates that a valid set of reference genes covering a broad expression range is recommended to accurately normalize and quantify the relative expression levels of the target gene(s) transcripts in many microbial processes. PMID- 19217563 TI - Identification of cyclophilin as a novel allergen from Platanus orientalis pollens by mass spectrometry. AB - Oriental plane trees are an important source of airborne allergens in cities of southwest Asia. In spite of extensive studies on Platanus acerifolia allergy, there are no reports on the molecular characterization of pollen allergens from Platanus orientalis trees. In this study, a newly recognized member of cyclophilin family with a molecular weight of 18 kDa was identified as being partly responsible for IgE reactivity of P. orientalis pollen extract. PMID- 19217564 TI - [Neonatal respiratory assistance: current trends]. PMID- 19217565 TI - [Subcutaneous gammaglobulin in common variable immunodeficiency. First experience in Spain]. AB - INTRODUCTION AND AIM: Weekly home-based subcutaneous immunoglobulin (SCIg) therapy is an alternative to intravenous immunoglobulin (IVIg) in the treatment of patients with primary antibody deficiencies. The objective of this study was to investigate the efficacy, safety, related quality of life and cost effectiveness of SCIg in our area. MATERIALS AND METHODS: Observational and descriptive study including paediatric patients with common variable immunodeficiency (CVID) receiving SCIg in our hospital (November 2006 to April 2008). Obtained data were compared with those from the last year with IVIg. RESULTS: Eleven patients with CVID were included. Median age was 15 years. The median trough serum IgG level was 622 mg/dl with IVIg. In patients in whom the SCIg dose was maintained or reduced compared to IVIg, the median trough serum IgG level was 850 mg/dl (p < 0.0005). Annual rate of infection was 2.22 per patient year, without significant differences to IVIg (p = 0.212). There were 58 treatment-related adverse events (AE) reported with SCIg (45 local AE and 13 systemic AE). The most frequent treatment-related adverse event was infusion-site reaction. Switching to home-based subcutaneous IgG treatment led to significant improvements in quality of life and substantial cost savings. CONCLUSIONS: We conclude that subcutaneous administration of 16% SCIg is a safe and cost effective alternative to IVIg for replacement therapy of primary antibody deficiencies. Median trough serum IgG levels were higher with SCIg. Local AE were common but mild and the incidence decreased over time. Quality of life is significantly improved. PMID- 19217566 TI - [Continuous glucose monitoring system in the screening of glucose disorders in cystic fibrosis]. AB - BACKGROUND: Diabetes mellitus (DM) is an increasing complication of cystic fibrosis (CF). It is associated with enhance morbidity. Continuous glucose monitoring system (CGMS) could detect glucose disorders earlier than other screening tests usually used. AIMS: To compare oral glucose tolerance test (OGTT), HbA(1c) and CGMS in patients with CF and recent disorders of glucose homeostasis and to analyse changes in nutritional status and/or pulmonary function. PATIENTS AND METHODS: Thirteen patients with CF (11-22 years, 7 males) were studied using OGTT, HbA(1c) and CGMS. All of them had newly diagnosed glucose disturbances. They were not receiving steroid therapy or had an underlying illness. In all subjects we compared: HbA(1c) levels (%), fasting and 2-hours glucose OGTT (mg/dl) and glucose CGMS values (overall, fasting, 2-hours post mean-meals and excursions >140mg/dl at any time). Furthermore, body mass index, forced expiratory volume in the first second (%) and forced vital capacity (%) were evaluated in the previous year and at the time of the study. We also analysed exocrine pancreatic function and CF-mutation. RESULTS: Mean age at diagnosis of glucose disturbance was 16.4 years. All patients had insufficient exocrine pancreatic function and 11/13 presented DeltaF508 CF-mutation. Only one patient was diagnosed with DM using OGGT and 7/13 (53.8%) with CGMS. A total 77% of patients had poor nutritional status and/or pulmonary function at time of diagnosing the glucose disorder. Only 4 patients had abnormal HbA(1c) levels. CONCLUSIONS: CGMS allows a better detection of glucose disorders than OGTT. Glucose homeostasis abnormalities are associated with a decrease in nutritional status and/or pulmonary function. HbA(1c) does not aid in the early diagnose of glucose disorders. PMID- 19217567 TI - [Optimising azathioprine treatment: determination of thiopurine methyltransferase activity and thiopurine metabolites]. AB - INTRODUCTION: Individualised doses of azathioprine (AZA) may be prescribed by monitoring the levels of the enzyme thiopurine methyltransferase (TPMT). The measurements of thiopurine metabolites of AZA, 6-thioguanine (6-TGN) and 6 methylmercaptopurine (6-MMP), have also been reported as new markers of AZA activity. OBJECTIVES: To describe TPMT phenotype in our population and to establish a relationship between thiopurine metabolites,and therapeutic activity and adverse effects. MATERIAL AND METHODS: Data on TPMT were retrospectively collected from 107 patients, and 6-TGN and 6-MMP levels in 18 patients currently on treatment with AZA (Crohn's disease 5, ulcerative colitis 5, autoimmune hepatitis 5). RESULTS: Mean value of TPMT was 20.19U/ml. None of the patients had a TPMT activity<5U/ml. Of the 18 patients on treatment, 13 showed sub-therapeutic levels of 6-TGN (<235pmol/8x10(8) red blood cells). Clinical remission was maintained in 45% of patients. Mean levels of 6-TGN in patients with clinical remission were 259pmol/8x10(8) red blood cells versus 209pmol/8x10(8) red blood cells in non-responders (p=0.37). There was an inverse relationship (r=-0.28) between TPMT and 6-TGN levels. Toxic effects occurred in 6 of 18 patients, with leukopenia in 5 and hyperamylasemia in 1. CONCLUSIONS: Determination of TPMT and monitoring of thiopurine metabolites allows AZA treatment to be optimised, although further studies are necessary to establish therapeutic effectiveness and toxicity ranges. PMID- 19217568 TI - [Acute ethanol intoxication in a paediatric emergency department]. AB - INTRODUCTION AND AIM: Patients arriving in Paediatric Emergency Departments due to alcohol abuse is fairly common. The aim of this study is to determine the profile of the patients seen in the emergency room for acute ethanol intoxication. MATERIAL AND METHOD: A descriptive study was performed on all patients seen in the emergency department with an acute ethanol intoxication over a 12-month period (2007). Patients with accidental poisoning or with related chronic illness were excluded. RESULTS: A total of 104 patients were included (50% males). Their mean age was 16.2 years (standar desviation [SD] 1.2 years); 95.8% were older than 14 years. There was on patient who was seen twice for the same reason. The majority, 72.4%, were seen on holiday days; 82.9%, from 8:00 pm to 8:00 am. A total of 81.9% of the patients were brought in by ambulance. No cases of hypoglycaemia were detected and 7.5% of the patients had hypothermia. Ethanol levels and urine toxicology were measured in 66 cases. The mean blood ethanol was 186.1mg/dl (SD 56.2mg/dl); 21.2% were slightly intoxicated, 75.8% moderate and 3% severe. Cannabis was detected in seven cases and amphetamines in three. Three patients needed hospital admission. CONCLUSIONS: The profile of the patient who arrives to emergency department with acute ethanol intoxication is a teenager who is seen on holiday nights, after a consumption of high proof alcoholic drinks, presents a moderate intoxication, and does not require hospitalisation. PMID- 19217569 TI - [Respiratory care in neonatal intensive care units. Situation in the year 2005]. AB - AIM: To learn the characteristic of the neonatal intensive care units (NICUs) that offer neonatal respiratory assistance in Spain. MATERIAL AND METHOD: A structured survey was developed and sent to all Spanish neonatal units to learn about the respiratory care offered in 2005. RESULTS: A total of 96 Units answered the survey, with an estimated representatively of 63%, with a range from 3 to 92%, depending on the geographical area. Level IIIc Units were in the upper range. Answer the survey 26 units type IIb (27%), 16 IIIa (17%), 40 IIIb (42%) and 14 IIIc (14%). The total number of level III NICU beds was 541 (1.2 beds per 1000 livebirths; range, 0.7-1.7). The mean number of beds per NICU was 4.1 in level IIIa Units, 2.8 in those IIIb and 14.6 in type IIIc NICUs. In level III NICUs, the bed per physician ratio was 2.4 and that of beds per registered nurse was 2.8 (2.2 in level IIIc NICUs). There were a total 13,219 admissions, 54% of those needed mechanical ventilation (36% in IIIa and 65% in level IIIc NICUs). Oxygen blenders for resuscitation at birth were available in 42% of level IIIb and IIIc NICUs. NICUs had one neonatal ventilator per bed, and 63% of units had high frequency ventilation available. All units had nasal-CPAP systems, 25% of level IIIa Units, 58% IIIb and 64% of those type IIIc had systems for nasal ventilation. All level IIIc and 93% of level IIIb NICUs were able to provide inhaled nitric oxygen therapy. Four NICUS offered ECMO. CONCLUSIONS: The mean number of NICU beds per 1000 livebirths is within the lower limits of those been recommended, and there were wide variations among different geographical areas. A 54% of those babies admitted to NICUs required mechanical ventilation. The mean number of NICU beds per registered nurse was 2.8. There was an adequate number of neonatal ventilators (one per bed) and 63% were able to provide HFV. All NICUs hand n-CPAP systems. PMID- 19217570 TI - [Neonatal and perinatal mortality in hospitals of the Basque Country-Navarre Neonatal Study Group (GEN-VN) during the period 2000-2006]. AB - BACKGROUND: In 2000, the Basque-Navarre Neonatal Study Group (GEN-VN) was created with a clear objective of studying the perinatal and neonatal health outcomes in newborns. OBJECTIVE: The aim of this investigation is to present the trends of neonatal and perinatal mortality and their causes in the hospitals of the Basque Navarre Neonatal Study Group from 2000 to 2006. PATIENTS AND METHOD: A descriptive study was carried out on the 157,623 births in the participating hospitals of the Basque-Navarre Neonatal Study Group, from 2000 to 2006. During this period, of the total births, 156.904 were born alive, there were 719 foetal deaths and 363 newborns died within the first 28 days of life. Perinatal and neonatal mortality was analysed, raw and stratified by gestational age and birth weight. RESULTS: The Basque -Navarre Neonatal Study Group collects from 80.5% to 96.8% of all the births reported by the authorities. There is a decreasing trend in foetal and perinatal mortality rates from 2000 to 2006. However, neonatal mortality rates shows a stable trend when compared with the descense in the last years of the 20th century. The most frequent causes of death are respiratory, infections and the congenital anomalies. CONCLUSIONS: Collecting regional data improves the quality of neonatal and perinatal mortality studies. Very low birth weight and very low gestational age newborns require special quality of care due their high mortality (54.8% of neonatal mortality in 2006), therefore further studies are required on the impact of these newborns on neonatal mortality in our hospitals. PMID- 19217571 TI - [Prenatal exposure to drugs of abuse using meconium analysis in a low socioeconomic population in Barcelona]. AB - INTRODUCTION: For the first time in Europe, the <> aimed to estimate the prevalence of drug use by pregnant women and the subsequent foetal exposure to illicit drugs. PATIENTS AND METHOD: Between October 2002 and February 2004, 1209 mother-infant dyads from the Hospital del Mar, Barcelona, Spain met eligibility criteria and agreed to participate in the study. Data on socio economic and demographic characteristics and on drug habits during pregnancy were collected using a structured questionnaire. Neonatal meconium was collected within 24h after birth and analyzed by standardized chromatographic techniques for the presence of opiates, cocaine, cannabinoids and amphetamines. RESULTS: Meconium analysis showed an overall 10.9% positivity for drugs of abuse, with a specific prevalence of heroin, cocaine and cannabis with foetal exposure of 4.7, 2.6 and 5.3%, respectively. Structured interviews also revealed that 0.3, 1.2 and 1.5% of mothers used heroin, cocaine and cannabis, respectively, while only one mother declared ecstasy consumption, confirmed by meconium analysis. Parental ethnicity and working class was not associated with drug use. Drug consuming mothers were shown to have a higher number of previous abortions when compared to non-consumer mothers, which was probably due to a lack of family planning. Significantly lower birth weight and length was found in newborns from mothers exposed to cocaine alone or in combination with other drugs. CONCLUSIONS: This study, although developed in a low socio-economic-status cohort, may serve as an eye opener for any hidden non-negligible drug consumption during pregnancy. In this sense, meconium analysis can be important to identify neonates with a high suspicion of exposure to drugs of abuse in utero, and provides the basis for appropriate treatment and adequate medical and social follow-up. PMID- 19217572 TI - [Neonatal complications associated to HELLP syndrome]. AB - INTRODUCTION: HELLP syndrome is a variant of pregnancy-induced hypertension that is associated with significant maternal and perinatal morbidity and mortality. The aim of our study was to investigate the neonatal complications associated to this syndrome. PATIENTS AND METHOD: A retrospective observational study was carried out on all newborns of mothers with HELLP syndrome in Virgen del Rocio hospital from 1995 to 2005. There were 120 newborns of 99 mothers with HELLP syndrome. Gestational age, birth weight, length, skull perimeter, number of hospital admissions and mortality were analyzed. The birth weight, length and skull perimeter were compared with a healthy population of the same gestational age using Lubchenco charts. The statistical relationships were determined between the mothers' platelet counts and the birth weight and perinatal mortality. RESULTS: A total of 80% of pregnancies were preterm delivery with a mean gestational age of 33 weeks. Mean birth weight was 1,834g, length 41cm and skull perimeter 29cm. A third of newborns had fetal growth restriction. 61% of newborns needed admitting to hospital due to prematurity and low birth weight. There were 24 perinatal deaths. We did not find any correlation between the number of platelets of the mother and birth weight or perinatal mortality. CONCLUSIONS: HELLP syndrome is an uncommon but potentially serious complication of pregnancy which is associated with an increased risk of adverse maternal and fetal outcome. PMID- 19217573 TI - [Griscelli-Prunieras syndrome: report of two cases]. AB - Griscelli-Prunieras syndrome (GS) is a rare autosomal recessive disorder characterized by partial albinism. His pathogenic mechanism is associated with defects in the packaging of melanin and other cellular proteins. GS is classified into 3 types based on the genetic and molecular features. Mutations in the genes which cause GS are known. We report two first cases described in Spain who presented a silver-gray sheen of the hair and a severe immune disorder. They were studied for mutations principally related to this syndrome. Two patients showed the Rab27a mutation (frequently associated with GS2). The natural disorder evolution differs considerably among the various forms, so a genetic study is essential in GS to achieve the most accurate prognosis and treatment possible. PMID- 19217574 TI - [Unusually high frequency of complications in acute otitis media]. AB - INTRODUCTION: Intracranial complications (ICC) secondary to otitis media are unusual but potentially life-threatening. We report an unusually high frequency of these events, and describe their clinical and epidemiological features. MATERIAL AND METHOD: A retrospective study of all pediatric patients with ICC admitted to our tertiary hospital from April 2004 to November 2007. RESULTS: Eight patients had ICC: Sigmoid sinus thrombosis (4), lateral sinus thrombosis (1), meningitis (2), epidural abscess (1), otitic hydrocephalus (1). Four of the patients had acute mastoiditis. Pre-admission oral antibiotics were administered in 87.5% of the patients. Fever, otalgia, headache and VI and VII cranial nerves paralysis were the most frequent symptoms associated. The microorganism could be isolated in 3 patients: S pyogenes (2) and P mirabilis (1); the remaining patients had negative cultures. All of the patients received broad-spectrum parenteral antibiotics and 2 of them underwent a radical mastoidectomy. CONCLUSIONS: We report an unusually high incidence of cranial complications secondary to acute otitis media. Multicenter studies are needed in order to assess whether there is an increase in the incidence of these events. PMID- 19217575 TI - [Recent advances in research on neonatal resuscitation: an update]. AB - Coinciding with the Annual Meeting of the Pediatric Academic Societies (PAS) last May 2008 in Honolulu (Hawaii, USA), Professor Colin Morley and his colleagues (University of Melbourne, Australia) held a Workshop on Research in Neonatal Resuscitation. Experts in the field presented their results and future projects over 2 days. The subjects presented during the workshop were: oxygen, air and oximetry; when and where to apply surfactant; neonatal resuscitation research techniques; teaching neonatal resuscitation; cellular physiology and biology; CPAP/PEEP/prolonged inspiration; video recording during resuscitation. Sessions were characterized by an interactive discussion. Our intention is to tell about some of the most innovative aspects that might interest our neonatal colleagues who did not have the opportunity to attend the meeting. As much of the information contained in this article is on experiments based, we recommend the reader not to consider it for immediate application in clinical practice until it has been validated by sufficient proof. PMID- 19217576 TI - [A guide to the clinical diagnosis and urgent treatment of neonatal hyperammonaemia]. AB - Symptomatic hyperammonaemia in newborn is a medical emergency that should be recognised in its early stages, specifically diagnosed and aggressively treated to improve the immediate and long-term prognosis of these children. The paediatrician and the neonatal doctor should have a diagnosis-therapy scheme for its urgent management. PMID- 19217578 TI - [n-3 and n-6 fatty acids in plasma at birth and at one year and the relationship with the type of feeding]. PMID- 19217579 TI - [Congenital syphilis in a 7-week old baby: could it be prevented with another protocol?]. PMID- 19217580 TI - [Basal ganglia infarction secondary to a Varicella-Zoster infection]. PMID- 19217581 TI - [Meropenem and valproic acid. An interaction to remember]. PMID- 19217582 TI - [Meat bolus impactation in a 12-year-old]. PMID- 19217583 TI - [Herpes esophagitis as a cause of acute dysphagia]. PMID- 19217584 TI - [Enteropathic-like acrodermatitis in propionic acidaemia]. PMID- 19217585 TI - [Micropenis and bilateral cryptorchidism secondary to vanishing testes syndrome]. PMID- 19217586 TI - [Spontaneous puberty and menarche in a patient with Turner syndrome and 45X monosomy]. PMID- 19217587 TI - [Systemic lupus erythematosus with fulminant neurological involvement]. PMID- 19217589 TI - Just how stable are stable aesthetic features? Symmetry, complexity, and the jaws of massive familiarization. AB - Using both group- and individual-level analyses, we explored the complex and dynamic effects of basic visual features on aesthetic judgment. Specifically, the mediating influence of familiarization on the combined effects of complexity and symmetry on aesthetic judgment was examined. Experiment 1 showed that symmetry and complexity are indeed powerful determinants of aesthetic judgment. Experiment 2 demonstrated that massive familiarization generated contrast effects for complexity: participants familiarized to simple stimuli subsequently judged complex stimuli more beautiful and participants familiarized to complex stimuli subsequently judged simple stimuli more beautiful. In contrast, moderate familiarization in Experiment 3 did not elicit the above effects. Group-level analyses were augmented with judgment analyses of individual response patterns resulting in a more comprehensive assessment of aesthetic judgment. PMID- 19217590 TI - Chorioamnionitis induced by intraamniotic lipopolysaccharide resulted in an interval-dependent increase in central nervous system injury in the fetal sheep. AB - OBJECTIVE: We quantified the impact of chorioamnionitis on both the white and gray matter structures of the preterm ovine central nervous system (CNS). STUDY DESIGN: The CNS was studied at 125 days of gestation, either 2 or 14 days after the intraamniotic administration of 10 mg of lipopolysaccharide (LPS) (Escherichia coli) or saline. Apoptotic cells and cell types were analyzed in the brain, cerebellum, and spinal cord using flow cytometry. RESULTS: Apoptosis and microglial activation increased in all regions with prolonged exposure to LPS induced chorioamnionitis. Astrocytes were increased in the brain and cerebellum of LPS-exposed fetuses but not in the spinal cord. Mature oligodendrocytes decreased in the cerebral and cerebellar white matter, the cerebral cortex, caudate putamen, and hippocampus 14 days after LPS. Neurons in the cerebral cortex, hippocampus, and substantia nigra were reduced 14 days after LPS. CONCLUSION: Fetal inflammation globally but differentially affected the CNS depending on the maturational stage of the brain region. PMID- 19217591 TI - Drug use and limited prenatal care: an examination of responsible barriers. AB - OBJECTIVE: The purpose of this study was to determine sociodemographic, psychosocial, and health belief factors that explain the association between maternal drug use and little or no prenatal care. STUDY DESIGN: A cohort of 812 low-income women who delivered at Johns Hopkins Hospital were administered a postpartum survey. Drug use was determined by self-report, medical record, and toxicologic screens. Medical records were abstracted to determine little or no prenatal care, as defined by 7 days, and 69 (49.3%) survived >30 days, whereas in group 2, respective survival was 110 of 130 (84.6%) and 86 of 130 (66.2%). Average cold ischemic times in the 2 groups were 48.5 +/- 5.1 minutes and 31 +/- 3.0 minutes, respectively. There was a significant difference between the 2 groups (P <.05). In most cases, the average volume of bleeding during recipient surgery was <1 mL using the simplified 3-cuff anastomosis technique. There was shorter graft revascularization time with the new model of sutureless microanastomosis using cuff apparatus for OIT in rats compared with the control group. The method adopted in group 2 was much easier, more stable, and more feasible than that in group 1. Sixty-three rats died in 7 days, and autopsy verified the causative factors leading to death, which are summarized in the text. The results obtained were acceptable and satisfactory. Overall, there was a comparative lower incidence of complications associated with the procedure used in group 2. CONCLUSIONS: The modified 3-cuff anastomosis technique for rat OIT models has several obvious advantages, which can be summarized as follows: vascular anastomosis is stable and simplified, and blood loss is significantly decreased; natural anatomic physiologic portal graft drainage is maintained; and intraoperative mortality and postsurgical morbidity are minimized. Furthermore, technical refinement of rat OIT models established by our research team can be carried out without a microscope and can be easily implemented in the laboratory by 1 trainee with acceptable success after a short period of training. We regard it as one of the best available orthotopic small bowel transplantation methods in rat. PMID- 19217612 TI - Differential performance on the go/no-go task as a function of the autogenous reactive taxonomy of obsessions: findings from a non-treatment seeking sample. AB - It has been suggested that obsessive-compulsive disorder is characterized by impaired inhibitory control. Response inhibition is a cognitive process required for one to cancel or suppress dominant but inappropriate responses. The present study examined response inhibition among non-treatment seeking individuals diagnosed with OCD and individuals with low levels of OCD symptoms using a computerized visual go/no-go task. Specifically, we sought to examine a prediction from the autogenous-reactive subtype model of obsessions (Lee, H.-J., & Kwon, S.-M. (2003). Two different types of obsession: autogenous obsessions and reactive obsessions. Behaviour Research and Therapy, 41, 11-29; Lee, H.-J., & Telch, M. J. (2008). Autogenous obsessions and reactive obsessions. In J. Abramowitz, S. Taylor, & D. McKay (Eds.), Obsessive-compulsive disorder: subtypes and spectrum conditions. New York: Elsevier., asserting that OCD individuals presenting with the autogenous subtype of obsessions will display greater difficulty in inhibitory control relative to individuals presenting with obsessions of the reactive subtype. Results showed that individuals with OCD of the autogenous subtype displayed more deficient inhibitory control on the visual go/no-go task as indexed by a longer response delay between the original stimuli set and the reversed stimuli set compared to individuals with OCD of the reactive subtype or individuals with low levels of OCD symptoms. PMID- 19217611 TI - Enamel mineralization in the absence of maturation stage ameloblasts. AB - The role of maturation stage ameloblasts is not clear yet. The aim of this study was to verify to which extent enamel mineralizes in the absence of these cells. Maturation stage ameloblasts and adjacent dental follicle cells from rat lower incisors were surgically removed and the limits of this removal were marked by notches made in the enamel. Histological analysis confirmed that the ameloblasts had been removed within the limits of the notches. The teeth erupted and when the notches appeared in the mouth, the enamel in the experimental teeth was hard but whitish compared to the yellowish colour of the contralateral incisors used as control. SEM images revealed similar enamel rod arrangement in both groups. Decreased mineral content was observed in some specimens by polarized light microscopy, and microhardness values were much lower in the experimental teeth. FTIR analysis showed that higher amounts of protein were found in most experimental teeth, compared with the control teeth. Enamel proteins could not be resolved on 15% SDS-PAGE gels, suggesting that most of them were below 5kDa. These results suggest that the enamel matured in the absence of ameloblasts has increased protein content and a much lower mineral content, suggesting that maturation stage ameloblasts are essential for proper enamel mineralization. PMID- 19217613 TI - Disorientation, confabulation, and extinction capacity: clues on how the brain creates reality. AB - BACKGROUND: Disorientation and confabulation often have a common course, independent of amnesia. Behaviorally spontaneous confabulation is the form in which patients act according to a false concept of reality; they fail to abandon action plans (anticipations) that do not pertain to the present situation. This continued enactment of previously valid but meanwhile invalidated anticipations can be conceived as deficient extinction capacity, that is, failure to integrate negative prediction errors into behavior. In this study, we explored whether disorientation and behaviorally spontaneous confabulation are associated with extinction failure. METHODS: Twenty-five patients hospitalized for neurorehabilitation after first-ever brain injury who either had severe amnesia (n = 17), an orbitofrontal lesion (n = 14), or both (n = 6) were tested regarding disorientation (questionnaire) and performed an experimental task of association learning and extinction. Five patients were also classified as behaviorally spontaneous confabulators. RESULTS: Extinction capacity explained 66% of the variance of orientation in the whole group of patients (amnesics only, 56%; orbitofrontal group only, 90%), whereas association learning explained only 17% of the variance in the whole group (amnesics only, 7%; orbitofrontal group only, 16%). Also, extinction capacity, but not association learning, significantly distinguished between behaviorally spontaneous confabulators and all other subjects. CONCLUSIONS: Disorientation and behaviorally spontaneous confabulation are strongly and specifically associated with a failure of extinction, the ability to learn that previously appropriate anticipations no longer apply. Rather than invoking high-level monitoring processes, the human brain seems to make use of an ancient biological faculty-extinction-to keep thought and behavior in phase with reality. PMID- 19217614 TI - Structural insight into the mechanism of streptozotocin inhibition of O GlcNAcase. AB - Despite decades of its use in diabetes research, the mechanism of cytotoxicity of streptozotocin (STZ) toward pancreatic beta-islet cells has remained a topic of discussion. Although STZ toxicity is likely a function of its capacity to promote DNA alkylation, it has been proposed that STZ induces pancreatic beta-cell death through O-GlcNAcase inhibition. In this report, we explore the binding mode of STZ to a close homolog of human O-GlcNAcase, BtGH84 from Bacteroides thetaiotaomicron. Our results show that STZ binds in the enzyme active site in its intact form, without the formation of a covalent adduct, consistent with solution studies on BtGH84 and human O-GlcNAcase, as well as with structural work on a homolog from Clostridium perfringens. The active site of the BtGH84 is considerably deformed upon STZ binding and as a result the catalytic machinery is expelled from the binding cavity. PMID- 19217615 TI - Fine structural properties of natural and synthetic glycogens. AB - Glycogen, highly branched (1-->4)(1-->6)-linked alpha-d-glucan, can be extracted from natural sources such as animal tissues or shellfish (natural source glycogen, NSG). Glycogen can also be synthesized in vitro from glucose-1 phosphate using the cooperative action of alpha-glucan phosphorylase (GP, EC 2.4.1.1) and branching enzyme (BE, EC 2.4.1.18), or from short-chain amylose by the cooperative action of BE and amylomaltase (AM, EC 2.4.1.25). It has been shown that enzymatically synthesized glycogen (ESG) has structural and physicochemical properties similar to those of NSG. In this study, the fine structures of ESG and NSG were analyzed using isoamylase and alpha-amylase. Isoamylase completely hydrolyzed the alpha-1,6 linkages of ESG and NSG. The unit chain distribution (distribution of degrees of polymerization (DP) of alpha-1,4 linked chains) of ESG was slightly narrower than that of NSG. alpha-Amylase treatment revealed that initial profiles of hydrolyses of ESG and NSG were almost the same: both glycogens were digested slowly, compared with starch. The final products from NSG by alpha-amylase hydrolysis were glucose, maltose, maltotriose, branched oligosaccharides with DP4, and highly branched macrodextrin molecules with molecular weights of up to 10,000. When ESG was digested with excess amounts of alpha-amylase, much larger macrodextrins (molecular weight>10(6)) were detected. In contrast, oligosaccharides with DP 4-7 could not be detected from ESG. These results suggest that the alpha-1,6 linkages in ESG molecules are more regularly distributed than those in NSG molecules. PMID- 19217616 TI - The logical syntax of number words: theory, acquisition and processing. AB - Recent work on the acquisition of number words has emphasized the importance of integrating linguistic and developmental perspectives [Musolino, J. (2004). The semantics and acquisition of number words: Integrating linguistic and developmental perspectives. Cognition93, 1-41; Papafragou, A., Musolino, J. (2003). Scalar implicatures: Scalar implicatures: Experiments at the semantics pragmatics interface. Cognition, 86, 253-282; Hurewitz, F., Papafragou, A., Gleitman, L., Gelman, R. (2006). Asymmetries in the acquisition of numbers and quantifiers. Language Learning and Development, 2, 76-97; Huang, Y. T., Snedeker, J., Spelke, L. (submitted for publication). What exactly do numbers mean?]. Specifically, these studies have shown that data from experimental investigations of child language can be used to illuminate core theoretical issues in the semantic and pragmatic analysis of number terms. In this article, I extend this approach to the logico-syntactic properties of number words, focusing on the way numerals interact with each other (e.g. Three boys are holding two balloons) as well as with other quantified expressions (e.g. Three boys are holding each balloon). On the basis of their intuitions, linguists have claimed that such sentences give rise to at least four different interpretations, reflecting the complexity of the linguistic structure and syntactic operations involved. Using psycholinguistic experimentation with preschoolers (n=32) and adult speakers of English (n=32), I show that (a) for adults, the intuitions of linguists can be verified experimentally, (b) by the age of 5, children have knowledge of the core aspects of the logical syntax of number words, (c) in spite of this knowledge, children nevertheless differ from adults in systematic ways, (d) the differences observed between children and adults can be accounted for on the basis of an independently motivated, linguistically-based processing model [Geurts, B. (2003). Quantifying kids. Language Acquisition, 11(4), 197-218]. In doing so, this work ties together research on the acquisition of the number vocabulary with a growing body of work on the development of quantification and sentence processing abilities in young children [Geurts, 2003; Lidz, J., Musolino, J. (2002). Children's command of quantification. Cognition, 84, 113-154; Musolino, J., Lidz, J. (2003). The scope of isomorphism: Turning adults into children. Language Acquisition, 11(4), 277-291; Trueswell, J., Sekerina, I., Hilland, N., Logrip, M. (1999). The kindergarten-path effect: Studying on-line sentence processing in young children. Cognition, 73, 89-134; Noveck, I. (2001). When children are more logical than adults: Experimental investigations of scalar implicature. Cognition, 78, 165-188; Noveck, I., Guelminger, R., Georgieff, N., & Labruyere, N. (2007). What autism can tell us about every. . . not sentences. Journal of Semantics,24(1), 73-90. On a more general level, this work confirms the importance of integrating formal and developmental perspectives [Musolino, 2004], this time by highlighting the explanatory power of linguistically-based models of language acquisition and by showing that the complex structure postulated by linguists has important implications for developmental accounts of the number vocabulary. PMID- 19217617 TI - Characterization of the similarity between diffusion tensors for image registration. AB - Registration algorithms are usually based on measures that must be optimized, whose choice has a great influence in the final result. For diffusion tensor imaging, scalar measures cannot be directly applied, and therefore new cost functions must be defined, regarding the special features of these data. We present a new pointwise similarity measure, named diffusion type based (DTB), that considers the physical meaning of the diffusion tensor. Theoretically, it is proved that DTB corrects the drawbacks of previous analogous measures, as well as it provides a more realistic result than generic measures. These conclusions are corroborated by experiments, where registration algorithms are driven by DTB and other broadly used measures. It is shown both quantitatively and visually that DTB leads to better results, and is more robust in presence of noise. PMID- 19217618 TI - Development of electronic software for the management of trauma patients on the orthopaedic unit. AB - INTRODUCTION: Continuity of patient care is an essential prerequisite for the successful running of a trauma surgery service. This is becoming increasingly difficult because of the new working arrangements of junior doctors. Handover is now central to ensure continuity of care following shift change over. The purpose of this study was to compare the quality of information handed over using the traditional ad hoc method of a handover sheet versus a web-based electronic software programme. It was hoped that through improved quality of handover the new system would have a positive impact on clinical care, risk and time management. METHODS: Data was prospectively collected and analyzed using the SPSS 14 statistical package. The handover data of 350 patients using a paper-based system was compared to the data of 357 cases using the web-based system. Key data included basic demographic data, responsible surgeon, location of patient, injury site including site, whether fractures were open or closed, concomitant injuries and the treatment plan. A survey was conducted amongst health care providers to assess the impact of the new software. RESULTS: With the introduction of the electronic handover system, patients with missing demographic data reduced from 35.1% to 0.8% (p<0.0001) and missing patient location from 18.6% to 3.6% (p<0.0001). Missing consultant information and missing diagnosis dropped from 12.9% to 2.0% (p<0.0001) and from 11.7% to 0.8% (p<0.0001), respectively. The missing information regarding side and anatomical site of the injury was reduced from 31.4% to 0.8% (p<0.0001) and from 13.7% to 1.1% (p<0.0001), respectively. In 96.6% of paper ad hoc handovers it was not stated whether the injury was 'closed' or 'open', whereas in the electronic group this information was evident in all 357 patients (p<0.0001). A treatment plan was included only in 52.3% of paper handovers compared to 94.7% (p<0.0001) of electronic handovers. A survey revealed 96% of members of the trauma team felt an improvement of handover since the introduction of the software, and 94% of members were satisfied with the software. CONCLUSIONS: The findings of our study show that the use of web-based electronic software is effective in facilitating and improving the quality of information passed during handover. Structured software also aids in improving work flow amongst the trauma team. We argue that an improvement in the quality of handover is an improvement in clinical practice. PMID- 19217619 TI - Outcomes of trauma in centenarians. AB - INTRODUCTION: We present the first study specifically investigating outcomes of centenarian (age>100 years) trauma patients. MATERIALS AND METHODS: We conducted a retrospective cohort study over a 2-year period using data from patients' notes. Inclusion criteria were: admission to Poole Hospital, age of 100 years old or greater and history of sustaining trauma. RESULTS: We used the hospital CaMIS database to identify patients aged 100 years and over who had incurred an injury. 24 patients met the entry criteria for the study: accounting for 26 admission episodes. 23 patients were female and 13 had sustained a fractured neck of femur. 10 patients underwent surgery and the rest were managed non-operatively. There were five inpatient deaths and the majority of the discharged patients returned to their original place of residence following treatment and rehabilitation. Overall mortality at 1 year was 10 out of 24. DISCUSSION: Our increasingly elderly population is leading to a growing burden upon the NHS. Admission of the extreme elderly trauma patient is becoming more commonplace and presents us with difficult management decisions. Orthopaedic/geriatric collaborative care is now the norm and should continue to develop and improve in the future. CONCLUSION: The extreme elderly should be afforded the same consideration for operative treatment as younger patients and that age alone should not be relied upon as a determinant of treatment and management in these patients. PMID- 19217620 TI - Exercise tolerance test for predicting coronary heart disease in asymptomatic individuals: A review. AB - In symptom-free subjects, exercise tolerance testing (ETT) has a doubtful utility for detecting latent coronary heart disease (CHD) because of frequent false positives, but it may be valuable for predicting future CHD. To clarify the latter question, we calculated CHD incidence associated with presence or absence of ETT-induced abnormalities of ST-segment depression, exercise capacity, and heart rate using published prospective ETT studies in primary prevention populations. Based on 5-23 years of follow-up, yearly incidence of fatal and non fatal CHD ranged from 0.9 to 5.8% in the presence of ST-segment depression and from 1.2 to 1.7% in the presence of impaired maximal heart rate, and pooling of all data provided a linear positive relationship between pre- and post-test CHD incidence. Yearly incidence of CHD death was 0.8% in the presence of ST-segment depression, 0.2-0.3% in the presence of impaired heart rate recovery, and 0.5% in the presence of low exercise capacity. Absence of ST-segment depression was associated with <1.5% yearly incidence of fatal and non-fatal CHD events, except for one study and <0.2% yearly incidence of CHD or CVD death in all populations analyzed. Lastly ETT-induced ST-segment depression conveys a CHD risk superior to that associated with ETT-induced heart rate and exercise capacity abnormalities. This may be due to difference in pretest CHD incidences in ETT studies. PMID- 19217621 TI - Atherosclerosis proceeds independently of thrombin-induced platelet activation in ApoE-/- mice. AB - Platelet activation has long been postulated to contribute to the development of atherosclerotic plaques, although the mechanism by which this might occur remains unknown. Thrombin is a potent platelet activator and transfusion of thrombin activated platelets into mice increases plaque formation, suggesting that thrombin-induced platelet activation might contribute to platelet-dependent atherosclerosis. Platelets from protease-activated receptor 4-deficient (Par4-/-) mice fail to respond to thrombin. To determine whether thrombin-activated platelets play a necessary role in a model of atherogenesis, we compared plaque formation and progression in Par4+/+ and Par4-/- mice in the atherosclerosis prone apolipoprotein E-deficient (ApoE-/-) background. Littermate Par4+/+ and Par4-/- mice, all ApoE-/-, were placed on a Western diet (21% fat, 0.15% cholesterol) for 5 or 10 weeks. The percent of aortic lumenal surface covered by plaques in Par4+/+ and Par4-/- mice was not different at either time point (2.2+/ 0.3% vs. 2.5+/-0.2% and 5.1+/-0.4% vs. 5.6+/-0.4% after 5 and 10 weeks, respectively). Further, no differences were detected in the cross-sectional area of plaques measured at the aortic root (1.53+/-0.17 vs. 1.66+/ 0.16x10(5)microm(2) and 12.56+/-1.23 vs. 13.03+/-0.55x10(5)microm(2) after 5 and 10 weeks, respectively). These findings indicate that thrombin-mediated platelet activation is not required for the early development of atherosclerotic plaques in the ApoE-/- mouse model and suggest that, if platelet activation is required for plaque formation under these experimental conditions, platelet activators other than thrombin suffice. PMID- 19217622 TI - Endothelin-1 activates ETA receptors on human vascular smooth muscle cells to yield proteoglycans with increased binding to LDL. AB - OBJECTIVE: Lipid retention in the vessel wall by glycosaminoglycan (GAG) chains on chondroitin/dermatan sulfate proteoglycans synthesized by vascular smooth muscle cells (VSMC) have recently been established as an early event in human coronary artery atherosclerosis. GAG structure can be altered by growth factors resulting in enhanced binding to low density lipoprotein (LDL). The aim of this study was to determine if proteoglycans produced by endothelin-1 treated VSMCs had increased binding to human LDL, to examine the effect of endothelin-1 on the synthesis and structure of proteoglycans and to elucidate the signalling pathway. METHODS AND RESULTS: Endothelin-1 stimulated an increase in [(35)S]sulfate and [(3)H]glucosamine incorporation into proteoglycans produced by human VSMC. The increase was due to an increase in GAG chain size assessed by SDS-PAGE and size exclusion chromatography. Increased radiolabel incorporation was inhibited by an ET(A) but not an ET(B) receptor antagonist. Endothelin-1 stimulated an increase in the 6:4 position sulfation ratio on the disaccharides of the GAG chains, an effect that was blocked by bosentan. The EGF receptor antagonist AG1478 did not affect the increase in GAG size mediated by endothelin-1. Inhibition of protein kinase C (PKC) with GF109203X or down regulation by PMA pre-treatment attenuated the effect of endothelin-1 on GAG synthesis. CONCLUSION: These data demonstrate that endothelin-1 stimulates changes in GAG chain structure that increase binding to LDL. This action of endothelin-1 may represent a new target for the prevention of lipid binding within the vascular wall and the associated complications resulting from this interaction. PMID- 19217623 TI - Cigarette smoking: an undertreated risk factor for cardiovascular disease. AB - Smoking and other forms of tobacco use are major risk factors for cardiovascular disease. The effect of cigarette smoking on cardiovascular health is evident even at the lowest levels of exposure. Yet, the adverse effects of smoking are reversible, with cardiovascular risk decreasing substantially within the first 2 years of smoking cessation. Significantly, the mortality from coronary heart disease is reduced more through smoking cessation than by other secondary preventive therapies such as cholesterol lowering. Smoking cessation is a highly effective way to improve cardiovascular health in smokers and extremely cost effective. However, smoking cessation therapies are not implemented maximally if they are implemented at all, perhaps because smoking is seen as a lifestyle choice or because smokers frequently relapse, as indicated by very low long-term quit rates. Too often, healthcare professionals, including lipidologists and cardiologists, do little to address their patients' smoking status, in spite of its impact on cardiovascular health. With the advent of new therapies to treat the nicotine addiction that results from smoking and other tobacco use, it is hoped that physicians will be more proactive in encouraging and implementing smoking cessation programs for their patients, with the goal of increasing long term quit rates, and reducing the morbidity and mortality associated with cardiovascular disease. PMID- 19217624 TI - Smoking cessation increases serum adiponectin levels in an apparently healthy Greek population. AB - BACKGROUND: Smoking has been associated with low serum levels of adiponectin, an adipocytokine with insulin-sensitizing, anti-inflammatory and anti-atherogenic properties. The objective of this study was to assess the early effect on adiponectin levels of smoking cessation supported by bupropion. METHODS: Apparently healthy smokers of both sexes with no additional cardiovascular risk factors were administered 150mg sustained-release bupropion twice daily for 9 weeks. Quitters constituted the active group and non-quitters the control group. Sandwich enzyme-linked immunosorbent assays were employed for the measurement of serum adiponectin and cotinine, the latter used for validation of self-reported abstinence. RESULTS: Among the 106 participants (mean age 44.5+/-11.3 years, 57 females, Brinkman index 512.2+/-98.4, basal adiponectin 7.2+/-1.5mg/L), 45 (42.5%) had quitted smoking at week 9. Quitters' post-cessation adiponectin levels were significantly increased (mean difference with baseline 1.9+/-0.8mg/L, 95% CI 1.2, 2.3; p<0.001), while non-quitters' adiponectin remained unaltered. A multiple regression model including female gender (standardized beta coefficient=0.480, p=0.002), age (0.355, p=0.003), body mass index (BMI) (-0.308, p=0.005), waist circumference (-0.276, p=0.008), smoking status (-0.255, p=0.010), and cotinine levels (-0.233, p=0.021) explained about two thirds of the variation in adiponectin levels (adjusted R(2)=0.656). CONCLUSIONS: Serum adiponectin levels appear to increase considerably within 2 months after smoking cessation. This finding may provide further insight into the mechanisms related to the detrimental effects of smoking and the benefits of quitting. PMID- 19217625 TI - Computational simulation of trabecular adaptation progress in human proximal femur during growth. AB - There are a large number of clinical and experimental studies that analyzed trabecular architecture as a result of bone adaptation. However, only a limited amount of quantitative data is currently available on the progress of trabecular adaptation during growth. In this paper, we proposed a two-step numerical simulation method that predicts trabecular adaptation progress during growth using a recently developed topology optimization algorithm, design space optimization (DSO), under the hypothesis that the mechanisms of DSO are functionally equivalent to those of bone adaptation. We applied the proposed scheme to trabecular adaptation simulation in human proximal femur. For the simulation, the full trabecular architecture in human proximal femur was represented by a two-dimensional microFE model with 50 microm resolution. In Step 1, we determined a reference value that regulates trabecular adaptation in human proximal femur. In Step 2, we simulated trabecular adaptation in human proximal femur during growth with the reference value derived in Step 1. We analyzed the architectural and mechanical properties of trabecular patterns through iterations. From the comparison with experimental data in the literature, we showed that in the early growth stage trabecular adaptation was achieved mainly by increasing bone volume fraction (or trabecular thickness), while in the later stage of the development the trabecular architecture gained higher structural efficiency by increasing structural anisotropy with a relatively low level of bone volume fraction (or trabecular thickness). We demonstrated that the proposed numerical framework predicted the growing progress of trabecular bone that has a close correlation with experimental data. PMID- 19217626 TI - Numerical study of the phase separation in binary lipid membrane containing protein inclusions under stationary shear flow. AB - The phase separation of lipids is believed to be responsible for the formation of lipid rafts in biological cell membrane. In the present work, a continuum model and a particle model are constructed to study the phase separation in binary lipid membrane containing inclusions under stationary shear flow. In each model, employing the cell dynamical system (CDS) approach, the kinetic equations of the confusion-advection process are numerically solved. Snapshot figures of the phase morphology are performed to intuitively display such phase evolving process. Considering the effects from both the inclusions and the shear flow, the time growth law of the characteristic domain size is discussed. PMID- 19217627 TI - 5th International Conference on Countercurrent Chromatography. Foreword. PMID- 19217628 TI - Analysis of recombinant human erythropoietin and novel erythropoiesis stimulating protein digests by immunoaffinity capillary electrophoresis-mass spectrometry. AB - In this work, we demonstrate that detection of a specific peptide marker by immunoaffinity capillary electrophoresis-mass spectrometry (IA-CE-MS) could be used to confirm the presence of recombinant human erythropoietin (rhEPO) in solution. Besides the carbohydrate content, the amino acid sequence of novel erythropoiesis stimulating protein (NESP) differs from human erythropoietin (hEPO) at five positions (Ala30Asn, His32Thr, Pro87Val, Trp88Asn, and Pro90Thr). After digesting both glycoproteins in solution by trypsin and PNGase F, two specific proteotypic peptides, EPO (77-97) and NESP (77-97) which differ in three amino acids, were selected as rhEPO and NESP markers, respectively. Both digests and their mixtures were analyzed by IA-CE-MS. The IA stationary phase was prepared from a custom made polyclonal anti-EPO (81-95) antibody immobilized on a solid support of CNBr-Sepharose 4B and was packed in a microcartridge near the inlet of the separation capillary. As the antibody was directed to a synthetic peptide EPO (81-95), only the proteotypic peptide EPO (77-97) was retained. The retained peptide was eluted, separated by electrophoresis and detected by MS. The method was specific to confirm the presence of rhEPO in solution. Although the limits of detection for the peptide marker were similar to those obtained with CE MS (a few mg/L), these results show the potential of this novel approach to detect in the future rhEPO and its analogues selectively and unambiguously at the levels expected in biological fluids. PMID- 19217629 TI - The effects of urea modification and heat treatment on the process of NO2 removal by wood-based activated carbon. AB - The removal of NO(2) on urea-modified and heat-treated wood-based activated carbons was studied. From the obtained results it was found that these modifications, especially when done at 950 degrees C, have a positive effect on NO(2) adsorption and on the retention of NO (the product of NO(2) reduction by carbon). The presence of moisture in the system enhances the removal of NO(2) but negatively affects the retention of NO. It is possible that the formation of active centers on the carbon surface and some increase in the volume of supermicropores during the high temperature treatment play a significant role in these removal processes. The surface of the carbons was analyzed in terms of the pK(a) distributions. The qualitative and quantitative analyses of the NO(2) adsorption products were carried out by means of FTIR and TA techniques, respectively. The main products found on the carbon surface were the NO(3) and NO(2) species. PMID- 19217630 TI - Electrostatic interaction between two cylindrical soft particles. AB - An expression for the potential energy of electrostatic interaction between two soft cylinders (i.e., polyelectrolyte-coated cylindrical particles) in an electrolyte solution is derived by applying Derjaguin's approximation to the corresponding interaction energy between two parallel soft plates for the case where the density of fixed charges within the polyelectrolyte layer is low. The interaction between two parallel cylinders and that between two crossed cylinders are considered. The obtained expression covers various limiting cases that include hard cylinder/hard cylinder interaction, cylindrical polyelectrolyte/cylindrical polyelectrolyte interaction, soft cylinder/cylindrical polyelectrolyte interaction, soft cylinder/hard cylinder interaction, and cylindrical polyelectrolyte/hard cylinder interaction. PMID- 19217631 TI - AA amyloidosis induced in sheep principally affects the gastrointestinal tract. AB - AA amyloidosis was initiated experimentally in adult sheep by induction of gangrenous pneumonia, an inflammatory process known to be associated with amyloid formation. A vegetable fragment contaminated with rumen content was instilled into the lungs of 4 experimental animals. A fifth animal was not inoculated and served as control. The animals were examined daily and blood and urine were sampled biweekly post-inoculation. One sheep was killed 18 days post-inoculation (dpi), another 49dpi, and the remaining two (as well as the control animal) 63dpi. Respiratory signs, diarrhoea and/or soft, unformed stool were observed in all inoculated sheep. All experimental animals developed gangrenous pneumonia with hypoalbuminaemia and hypergammaglobulinaemia, and elevated urinary protein, creatinine, gamma glutamyl transferase and ss-glucuronidase. Amyloid deposition was most pronounced in the gastrointestinal tract and was evident from 18dpi. Amyloid was present from the tongue to the rectum, but was most prominent in the duodenum where the deposits disrupted the normal mucosal architecture. Other body organs had only mild amyloid deposition. Immunohistochemistry confirmed that the deposits were AA amyloid. These findings suggest that the gastrointestinal tract is the main target organ for AA amyloid deposition in sheep. The observations in this experimental model must now be confirmed in animals with spontaneously arising AA amyloidosis. PMID- 19217632 TI - The effect of methylphenidate on postural stability under single and dual task conditions in children with attention deficit hyperactivity disorder - a double blind randomized control trial. AB - OBJECTIVES: To investigate the effects of Methylphenidate (MPH) on postural stability in attention deficit hyperactivity disorder (ADHD) children in single and dual task conditions. METHODS: A randomized controlled double-blind study analyzing postural stability in 24 ADHD children before and after MPH vs. placebo treatments, in three task conditions: (1) Single task, standing still; (2) dual task, standing still performing a memory-attention demanding task; (3) standing still listening to music. RESULTS: MPH resulted in a significant improvement in postural stability during the dual task condition and while listening to music, with no equivalent improvement in placebo controls. CONCLUSIONS: MPH improves postural stability in ADHD, especially when an additional task is performed. This is probably due to enhanced attention abilities, thus contributing to improved balance control during performance of tasks that require attention. MPH remains to be studied as a potential drug treatment to improve balance control and physical functioning in other clinical populations. PMID- 19217633 TI - Triterpenoids with antimicrobial activity from Drypetes inaequalis. AB - The air-dried stems and ripe fruit of Drypetes inaequalis Hutch. (Euphorbiaceae) were studied. Four triterpene derivatives, characterized as lup-20(29)-en 3beta,6alpha-diol, 3beta-acetoxylup-20(29)-en-6alpha-ol, 3beta-caffeoyloxylup 20(29)-en-6alpha-ol and 28-betad-glucopyranosyl-30-methyl 3beta-hydroxyolean-12 en-28,30-dioate along with 10 known compounds were isolated from the whole stems. One triterpene, characterized as 3alpha-hydroxyfriedelan-25-al along with six known compounds were isolated from the ripe fruit. Their structures were established on the basis of spectroscopic analysis and chemical evidence. The triterpenes were tested for antimicrobial activity against some Gram-positive and Gram-negative bacteria, and two of them appeared to be modestly active. PMID- 19217634 TI - Substrate specificity of plant UDP-dependent glycosyltransferases predicted from crystal structures and homology modeling. AB - Plant family 1 UDP-dependent glycosyltransferases (UGTs) catalyze the glycosylation of a plethora of bioactive natural products. In Arabidopsis thaliana, 120 UGT encoding genes have been identified. The crystal-based 3D structures of four plant UGTs have recently been published. Despite low sequence conservation, the UGTs show a highly conserved secondary and tertiary structure. The sugar acceptor and sugar donor substrates of UGTs are accommodated in the cleft formed between the N- and C-terminal domains. Several regions of the primary sequence contribute to the formation of the substrate binding pocket including structurally conserved domains as well as loop regions differing both with respect to their amino acid sequence and sequence length. In this review we provide a detailed analysis of the available plant UGT crystal structures to reveal structural features determining substrate specificity. The high 3D structural conservation of the plant UGTs render homology modeling an attractive tool for structure elucidation. The accuracy and utility of UGT structures obtained by homology modeling are discussed and quantitative assessments of model quality are performed by modeling of a plant UGT for which the 3D crystal structure is known. We conclude that homology modeling offers a high degree of accuracy. Shortcomings in homology modeling are also apparent with modeling of loop regions remaining as a particularly difficult task. PMID- 19217635 TI - [Differential diagnosis of status epilepticus]. AB - The diagnosis of status epilepticus can be retained, wrongly, in several circumstances. Nonepileptic pseudoseizures from a psychiatric origin and some movement disorders can mimic convulsive status epilepticus. Encephalopathy of various causes (post-anoxic, metabolic, toxic, Creutzfeldt-Jakob disease) can be wrongly taken for non-convulsive status epilepticus, mainly due to inadequate interpretation of the electroencephalogram (EEG). In these encephalopathies, the existence of (non-epileptic) myoclonus and the abolition of the EEG abnormalities with the use of a benzodiazepine (without correction of the clinical symptoms) are additional confounding factors, leading to false diagnosis. Nevertheless, in general, the diagnosis of status epilepticus can be confirmed or rejected base on a combined analysis of the clinical data and the EEG. PMID- 19217636 TI - Measuring derived acoustic power of an ultrasound surgical device in the linear and nonlinear operating modes. AB - OBJECTIVE AND MOTIVATION: The method for measuring derived acoustic power of an ultrasound point source in the form of a sonotrode tip has been considered in the free acoustic field, according to the IEC 61847 standard. The main objective of this work is measuring averaged pressure magnitude spatial distribution of an sonotrode tip in the free acoustic field conditions at different electrical excitation levels and calculation of the derived acoustic power at excitation frequency (f0 approximately 25 kHz). Finding the derived acoustic power of an ultrasonic surgical device in the strong cavitation regime of working, even in the considered laboratory conditions (anechoic pool), will enable better understanding of the biological effects on the tissue produced during operation with the considered device. EXPERIMENTAL METHOD: The pressure magnitude spatial distribution is measured using B&K 8103 hydrophone connected with a B&K 2626 conditioning amplifier, digital storage oscilloscope LeCroy Waverunner 474, where pressure waveforms in the field points are recorded. Using MATLAB with DSP processing toolbox, averaged power spectrum density of recorded pressure signals in different field positions is calculated. The measured pressure magnitude spatial distributions are fitted with the appropriate theoretical models. THEORETICAL APPROACHES: In the linear operating mode, using the acoustic reciprocity principle, the sonotrode tip is theoretically described as radially oscillating sphere (ROS) and transversely oscillating sphere (TOS) in the vicinity of pressure release boundary. The measured pressure magnitude spatial distribution is fitted with theoretical curves, describing the pressure field of the considered theoretical models. The velocity and displacement magnitudes with derived acoustic power of equivalent theoretical sources are found, and the electroacoustic efficiency factor is calculated. When the transmitter is excited at higher electrical power levels, the displacement magnitude of sonotrode tip is increased, and nonlinear behaviour in loading medium appears, with strong cavitation activity produced hydrodynamically. The presence of harmonics, subharmonics and ultraharmonics as a consequence of stable cavitation is evident in the averaged power spectral density. The cavitation noise with continuous frequency components is present as a consequence of transient cavitation. The averaged pressure magnitude at the frequency components of interest (discrete and continuous) in the field points is found by calculating average power spectral density of the recorded pressure waveform signal using the welch method. The frequency band of interest where average power spectral density is calculated is in the range from 15 Hz up to 120 kHz due to measurement system restrictions. The novelty in the approach is the application of the acoustic reciprocity principle on the nonlinear system (sonotrode tip and bubble cloud) to find necessary acoustic power of the equivalent acoustic source to produce the measured pressure magnitude in the field points at the frequency components of interest. RESULTS: In the nonlinear operating mode, the ROS model for the considered sonotrode tip is chosen due to the better agreement between measurement results and theoretical considerations. At higher excitation levels, it is shown that the averaged pressure magnitude spatial distribution of discrete frequency components, produced due to stable cavitation, can be fitted in the far field with the inverse distance law. The reduced electroacoustic efficiency factor, calculated at excitation frequency component as ratio of derived acoustic power with applied electrical power, is reduced from 40% in the linear to 3% in the strong nonlinear operating mode. The derived acoustic power at other frequency components (subharmonic, harmonic and ultraharmonic) is negligible in comparison with the derived acoustic power at excitation frequency. DISCUSSION AND CONCLUSIONS: The sonotrode tip and loading medium are shown in the strong cavitation regime as the coupled nonlinear dynamical system radiating acoustic power at frequency components appearing in the spectrum. The bubble cloud in the strong nonlinear operating mode decreases the derived acoustic power significantly at the excitation frequency. PMID- 19217637 TI - Lethal synergy of solar UV-radiation and H(2)O(2) on wild Fusarium solani spores in distilled and natural well water. AB - Environmentally-friendly disinfection methods are needed in many industrial applications. As a natural metabolite of many organisms, hydrogen peroxide (H(2)O(2))-based disinfection may be such a method as long as H(2)O(2) is used in non-toxic concentrations. Nevertheless, when applied alone as a disinfectant, H(2)O(2) concentrations need to be high enough to achieve significant pathogen reduction, and this may lead to phytotoxicity. This paper shows how H(2)O(2) disinfection concentrations could be significantly reduced by using the synergic lethality of H(2)O(2) and sunlight the first time for fungi and disinfection. Experiments were performed on spores of Fusarium solani, the ubiquitous, pytho- and human pathogenic fungus. Laboratory (250-mL bottles) and pilot plant solar reactors (2 x 14 L compound parabolic collectors, CPCs) were employed with distilled water and real well water under natural sunlight. This opens the way to applications for agricultural water resources, seed disinfection, curing of fungal skin infections, etc. PMID- 19217638 TI - Oxygen transfer in membrane bioreactors treating synthetic greywater. AB - Mass transfer coefficients (k(L)a) were studied in two pilot scale membrane bioreactors (MBR) with different setup configurations treating 200L/h of synthetic greywater with mixed liquor suspended solids' (MLSS) concentrations ranging from 4.7 to 19.5g/L. Besides the MLSS concentration, mixed liquor volatile suspended solids (MLVSS), total solids (TS), volatile solids (VS), chemical oxygen demand (COD) and anionic surfactants of the sludge were measured. Although the pilot plants differed essentially in their configurations and aeration systems, similar alpha-factors at the same MLSS concentration could be determined. A comparison of the results to the published values of other authors showed that not the MLSS concentration but rather the MLVSS concentration seems to be the decisive parameter which influences the oxygen transfer in activated sludge systems operating at a high sludge retention time (SRT). PMID- 19217639 TI - Enhanced washing of HCB contaminated soils by methyl-beta-cyclodextrin combined with ethanol. AB - The present study investigates the combined effect of cyclodextrin (CD) and cosolvent on the washing of hexachlorobenzene (HCB) from contaminated soils. Methyl-beta-cyclodextrin (MCD), ethanol, a natural soil (NS) and kaolin, were selected. Batch equilibrium experiments were conducted to quantify the performance of HCB solubilization and recovery by varied MCD/ethanol systems. Results show that with 30% or 50% of added ethanol, considerable synergistic effect on HCB solubilization was achieved in MCD/ethanol system. The formation of ternary MCD/ethanol/HCB complex is most likely responsible for the synergy. Similar synergy was further evidenced in the washing of HCB from kaolin and NS by MCD/ethanol system containing 30% ethanol. Unexpectedly, the combination of MCD and 50% ethanol exhibited negligible synergy of HCB recovery. The failure in promotion is probably due to a nonextractable fraction of HCB for NS, or a nearly complete HCB recovery from kaolin by 50% ethanol alone. Results also indicate that the synergistic effect of the MCD/ethanol system will be significantly hindered by a higher organic content and longer aging of the soil. Generally, our study suggests that compared to conventional CD or cosolvent-aided soil remediation, the combined use of CD and cosolvents would offer great superiority to the contaminant recovery. PMID- 19217640 TI - Phytotoxicity and bioavailability of cobalt to plants in a range of soils. AB - Risk assessments of existing substances, including metals, in the environment are being carried in the European Union, which require estimation of the predicted no effect concentration (PNEC) derived from biological toxicity assays. Few toxicity data exist for Co and higher plants, and the influence of soil properties on Co toxicity also needs to be evaluated systematically. We investigated phytotoxicity and bioavailability of Co to barley (Hordeum vulgare L.), oilseed rape (OSR; Brassica napus L.) and tomato (Lycopersicon esculentum L.) shoot growth in ten soils varying widely in soil properties using a standardised shoot biomass assay. The effective concentration of added Co causing 50% inhibition (EC(50)) ranged from 40 to 1708 mg kg(-1), from 7 to 966 mg kg(-1) and from 7 to 733 mg kg(-1) for the barley, OSR and tomato test, respectively, representing 43-, 138-, and 105-fold variation among soils. The EC(50) based on the Co concentration in soil solution varied less among soils (4-15 fold) than that based on the total added Co, suggesting that solubility of Co is a key factor influencing its toxicity to plants. Regressions of soil Co toxicity threshold values with various soil properties, showed that exchangeable calcium (Ca) concentration was the most consistent single predictor. For risk assessment, we suggest that Co toxicity threshold values for plants be normalised using the soil exchangeable Ca. PMID- 19217641 TI - PBDEs in US and German clothes dryer lint: a potential source of indoor contamination and exposure. AB - Polybrominated diphenyl ethers (PBDEs) are emerging persistent organic pollutants (POPs). Since the 1970s, PBDEs have been widely used as additive flame retardants in furniture and electronic equipment. Due to their wide use and persistent nature, these chemicals are found in the environment, human blood, breast milk and other tissues in increasing levels in recent decades. PBDEs are similar to polychlorinated biphenyls (PCBs) in structure and toxicity. However, unlike PCBs and PCDDs/PCDFs, the route of PBDE exposure is not almost exclusively through food. PBDE levels in US food are not markedly higher than in Europe, although US human blood and milk levels are an order of magnitude higher. For these reasons, other possible routes of PBDE exposure have been investigated to understand PBDE intake into humans. PBDE contaminated clothes dryer lint and household dust are indicators of indoor contamination and may be sources of human exposure through hand-to-mouth contact or dermal absorption. There are very few publications about PBDEs in US or European lint. Household dryer lint from 12 US and seven German homes were analyzed for PBDEs by gas chromatography coupled to high-resolution mass spectrometry. We found the median US total PBDE levels were more than 10 times higher than median German levels and the mean US levels were two times higher than mean German levels. The US levels ranged from 321 to 3073ngg(-1) (median: 803ngg(-1), mean: 1138ngg(-1)) and the German levels were from 330 to 2069ngg(-1) (median: 71ngg(-1), mean: 361ngg(-1)). PBDE contamination of lint was found in all samples; the source of the PBDEs may be from dryer electrical components and/or dust deposition onto clothing. PMID- 19217642 TI - A dynamic model for predicting chemical concentrations in water and biota during the planning phase of aquatic ecotoxicological tests. AB - An unsteady-state fugacity model has been developed and validated as a predictive tool that will be useful in the planning phase of aquatic ecotoxicological tests. The model predicts the compound concentration trends in water and biota in experimental aquaria, with respect to the chemical and experimental conditions. The model has been validated with two echinoderm species, Paracentrotus lividus and Antedon mediterranea after a 28-days exposure to p,p'-DDE or triphenyltin chloride (TPT-Cl), respectively. Differences between the predicted vs. measured concentrations of these compounds in water and biota were generally below a factor of two for both compounds. The model here proposed considers three different compartments, water, animals, and glass, and five loss processes: volatilisation, glass adsorption, abiotic degradation, bioconcentration and biotransformation. In particular, adsorption onto glass materials was introduced into the model by means of two equations (R(2) values of 0.86 and 0.90) relating the adsorption rate constant and glass-water partition coefficient on the base of the physical-chemical properties of the compound (log K(ow)). The model can be applied during the planning phase of ecotoxicological tests and for understanding the behaviour of the compound at this micro-ecosystem scale after the tests have been performed. PMID- 19217643 TI - Analysis of the dissipation kinetics of ivermectin at different temperatures and in four different soils. AB - The study target was to assess the usefulness of the OECD test guideline 307 for the veterinary pharmaceutical ivermectin. Laboratory microcosm studies were conducted to investigate the aerobic and anaerobic transformation of ivermectin in soils from three locations in Europe (York, Madrid and Tastrup) and an artificial soil. The reason to include an artificial soil in the study was to understand the exposure potential of ivermectin in a parallel eco-toxicological study with non-target organisms in this soil for a longer duration. Three kinetic models (first-order (SFO), availability-adjusted first-order (AAFO) and bi exponential first-order (BFO)) were applied to fit the observed transformation dynamics and to derive dissipation times. Dissipation rates were highly dependent on the tested soils. Under aerobic conditions, dissipation was remarkably faster in the three natural soils tested (DT(50)=16.1-36.1d) than in the artificial soil (DT(50)>500d). Furthermore, a clear increase in DT(50) values was seen when the temperature was lowered from 20 to 6 degrees C. The results indicated that dissipation in soils with comparably strong sorption and low degrees of desorption (i.e. the York soil and to some extent the Tastrup soil) were best described by the AAFO model. While dissipation in the Madrid soil which had a lower sorption coefficient and a higher degree of reversibility of sorption could be satisfactorily described with the SFO model. Our data further showed that no significant dissipation occurred under anaerobic conditions. PMID- 19217644 TI - Treating landfill leachate using passive aeration trickling filters; effects of leachate characteristics and temperature on rates and process dynamics. AB - Biological ammoniacal-nitrogen (NH(4)(+)-N) and organic carbon (TOC) treatment was investigated in replicated mesoscale attached microbial film trickling filters, treating strong and weak strength landfill leachates in batch mode at temperatures of 3, 10, 15 and 30 degrees C. Comparing leachates, rates of NH(4)(+)-N reduction (0.126-0.159 g m(-2) d(-1)) were predominantly unaffected by leachate characteristics; there were significant differences in TOC rates (0.072 0.194 g m(-2) d(-1)) but no trend relating to leachate strength. Rates of total oxidised nitrogen (TON) accumulation (0.012-0.144 g m(-2) d(-1)) were slower for strong leachates. Comparing temperatures, treatment rates varied between 0.029 0.319 g NH(4)(+)-N m(-2) d(-1) and 0.033-0.251 g C m(-2) d(-1) generally increasing with rising temperatures; rates at 3 degrees C were 9 and 13% of those at 30 degrees C for NH(4)(+)-N and TOC respectively. For the weak leachates (NH(4)(+)-N<140 mg l(-1)) complete oxidation of NH(4)(+)-N was achieved. For the strong leachates (NH(4)(+)-N 883-1150 mg l(-1)) a biphasic treatment response resulted in NH(4)(+)-N removal efficiencies of between 68 and 88% and for one leachate no direct transformation of NH(4)(+)-N to TON in bulk leachate. The temporal decoupling of NH(4)(+)-N oxidation and TON accumulation in this leachate could not be fully explained by denitrification, volatilisation or anammox, suggesting temporary storage of N within the treatment system. This study demonstrates that passive aeration trickling filters can treat well-buffered high NH(4)(+)-N strength landfill leachates under a range of temperatures and that leachate strength has no effect on initial NH(4)(+)-N treatment rates. Whether this approach is a practicable option depends on a range of site specific factors. PMID- 19217645 TI - Environmental fate of tungsten from military use. AB - This manuscript describes the distribution, fate and transport of tungsten used in training rounds at three small arms ranges at Camp Edwards on the Massachusetts Military Reservation (MMR), USA. Practice with tungsten/nylon rounds began in 2000 subsequent to a 1997 US Environmental Protection Agency ban on training with lead. Training with the tungsten rounds was halted in 2005 because of concerns regarding tungsten's environmental mobility and potential toxicity. This study, therefore, examines how tungsten partitions in the environment when fired on a small arms training range. Soil sampling revealed surface soil concentrations, highest at the berm face, up to 2080 mg/kg. Concentrations decreased rapidly with depth--at least by an order of magnitude by 25 cm. Nonetheless, tungsten concentrations remained above background to at least 150 cm. Pore-water samples from lysimeters installed in berm areas revealed a range of concentrations (<1-400 mg/L) elevated with respect to background although there was no discernable trend with depth. Groundwater monitoring well samples collected approximately 30 m below ground surface showed tungsten (0.001 0.56 mg/L) attributable to range use. PMID- 19217646 TI - Feather mercury concentrations and physiological condition of great egret and white ibis nestlings in the Florida Everglades. AB - Mercury contamination in the Florida Everglades has reportedly played a role in the recent decline of wading birds, although no studies have identified a mechanism leading to population-level effects. We assessed feather mercury levels in great egret (Ardea alba; n=91) and white ibis (Eudocimus albus; n=46) nestlings at breeding colonies in the Florida Everglades during a year (2006) with excellent breeding conditions (characterized by hydrology leading to concentrated prey) and a year with below average breeding conditions (2007). We also assessed the physiological condition of those nestlings based on levels of plasma and fecal corticosterone metabolites, and stress proteins 60 and 70. Mercury levels were higher in both species during the good breeding condition year (great egret=6.25 microg/g+/-0.81 SE, white ibis=1.47 microg/g+/-0.41 SE) and lower in the below average breeding year (great egret=1.60 microg/g+/-0.11 SE, white ibis=0.20 microg/g+/-0.03 SE). Nestlings were in better physiological condition in 2006, the year with higher feather mercury levels. These results support the hypothesis that nestlings are protected from the harmful effects of mercury through deposition of mercury in growing feathers. We found evidence to suggest shifts in diets of the two species, as a function of prey availability, thus altering their exposure profiles. However, we found no evidence to suggest they respond differently to mercury exposure. PMID- 19217647 TI - Arsenic levels in wipe samples collected from play structures constructed with CCA-treated wood: impact on exposure estimates. AB - Lumber treated with chromated copper arsenate (CCA) has been used in residential outdoor wood structures and playgrounds. The U.S. EPA has conducted a probabilistic assessment of children's exposure to arsenic from CCA-treated structures using the Stochastic Human Exposure and Dose Simulation model for the wood preservative scenario (SHEDS-Wood). The EPA assessment relied on data from an experimental study using adult volunteers and designed to measure arsenic in maximum hand and wipe loadings. Analyses using arsenic handloading data from a study of children playing on CCA-treated play structures in Edmonton, Canada, indicate that the maximum handloading values significantly overestimate the exposure that occurs during actual play. The objective of our paper is to assess whether the dislodgeable arsenic residues from structures in the Edmonton study are comparable to those observed in other studies and whether they support the conclusion that the values derived by EPA using modeled maximum loading values overestimate hand exposures. We compared dislodgeable arsenic residue data from structures in the playgrounds in the Edmonton study to levels observed in studies used in EPA's assessment. Our analysis showed that the dislodgeable arsenic levels in the Edmonton playground structures are similar to those in the studies used by EPA. Hence, the exposure estimates derived using the handloading data from children playing on CCA-treated structures are more representative of children's actual exposures than the overestimates derived by EPA using modeled maximum values. Handloading data from children playing on CCA-treated structures should be used to reduce the uncertainty of modeled estimates derived using the SHEDS-Wood model. PMID- 19217648 TI - Aortitis and spondyloarthritis--an unusual presentation: case report and review of the literature. AB - OBJECTIVES: To report a patient with aortic regurgitation (AR) and aortitis, who failed to respond to multiple attempts at surgical replacement of the affected valve and adjacent aorta, and in whom a late diagnosis of spondyloarthritis (SpA) was made. The relevant literature on the association of SpA and aortitis is reviewed. METHODS: Descriptive case report of a patient with AR secondary to aortic dilation and aortitis with a late diagnosis of SpA, and a review of the relevant literature (PubMed search 1956 through April 2008). RESULTS: A 36-year old man was admitted to the hospital for replacement of his aortic valve and aortic valve root as a result of AR after a dehiscence of a mechanical prosthetic valve and subsequent aortic pseudoaneurysm. His past medical history included 2 prior aortic valve replacements that failed due to late postoperative dehiscence. Histological findings from the aortic wall after the third operation showed signs of aortitis. A rheumatologic workup revealed a 6-year history of mild low back pain, radiological evidence of bilateral sacroiliitis, and presence of HLA-B27. A diagnosis of SpA, most likely ankylosing spondylitis (AS), was made. On reviewing the literature, AR has been diagnosed in 2 to 12% of the patients with AS, most of whom had long-lasting disease. There have been several reports of uncommon presentations of acute AR in young patients with juvenile AS; however, no similar adult case was found. CONCLUSION: This unusual case emphasizes the importance of early and adequate workup for SpA in young patients with lone AR or aortic dilation. PMID- 19217649 TI - The interface of pain and mood disturbances in the rheumatic diseases. AB - OBJECTIVES: To review the overlap of pain and depression in the rheumatic diseases, focusing on fibromyalgia (FM) and rheumatoid arthritis, and to provide treatment recommendations based on an understanding of this interface. METHODS: A literature search was performed through PubMed and Medline, for the years 1978 to 2008 and using the keywords: depression, mood disorders, pain, arthritis, fibromyalgia, rheumatoid arthritis, osteoarthritis, systemic lupus erythematosus, scleroderma, antidepressants, exercise, and cognitive behavioral therapy. RESULTS: This review focuses on the pain/depression interface in common rheumatic disorders. FM is an excellent model to study the proposed mechanisms of such interactions. The potential genetic, inflammatory/immune, morphologic, and neurohormonal mechanisms that have been reported in FM are reviewed and compared with similar studies in depression. The importance of the interface of pain and depression has been noted in rheumatoid arthritis and other rheumatic illnesses, although there are very few studies to investigate their pathophysiologic underpinnings. General therapeutic recommendations can be made based on these overlapping models of the interface of pain and depression. CONCLUSIONS: Clinical and pathophysiologic studies in FM demonstrate the striking overlap of pain and depression. These studies pertain to a better understanding of the pain/depression interface in all chronic rheumatic illnesses. PMID- 19217650 TI - Is remitting seronegative symmetrical synovitis with pitting edema (RS3PE) a subset of rheumatoid arthritis? AB - OBJECTIVES: To contrast and compare the spectrum of remitting seronegative symmetrical synovitis with pitting edema (RS3PE) with rheumatoid arthritis (RA) using an illustrative case. METHODS: The relevant English literature of RS3PE was searched using the keywords "RS3PE" alone and in combination with terms such as neoplasia and rheumatic disease. Original and review articles were reviewed and the clinical setting was exemplified with a case report. RESULTS: RS3PE initially was reported to represent a form of RA. However, RS3PE has clinical features that are different from both early- and late-onset RA, such as lack of bony erosions and rheumatoid factor. RS3PE is thought to involve vascular endothelial growth factor, suggesting an infectious etiology, generally has an excellent prognosis, and is associated with neoplasia not commonly seen in RA, and the RA associated human leukocyte antigen (HLA) DRB1 genotype is absent. CONCLUSIONS: Based on the clinical, laboratory, suspected infectious etiology, genetic differences, and types of associated malignancies, RS3PE appears to be a distinct entity rather than a subset of RA. PMID- 19217652 TI - Effects of methylene blue in reducing cholestatic oxidative stress and hepatic damage after bile-duct ligation in rats. AB - The aim of this study was to evaluate the effects of methylene blue against cholestatic oxidative stress and liver damage after ligation of the common bile duct in male Wistar rats. Eight animals were included in each of the following five groups: untreated control, methylene blue control, sham-operated, bile-duct ligation, and bile-duct ligation plus methylene blue. Methylene blue was administered intraperitoneally for 14 days at a daily dose of 2mg/kg per day. All rats were sacrificed 2 weeks following the experimental treatment and the livers of all groups were examined biochemically and histopathologically. The severity of cholestasis and hepatic injury were determined by changes in the plasma, including enzymatic activities: aspartate aminotransferase, alanine aminotransferase, gamma glutamine transferase, and also bilirubin levels. Malondialdehyde, nitric oxide and superoxide dismutase were measured to indicate the oxidative status in the liver tissue. Myeloperoxidase activity and levels of tissue hydroxyproline were determined as measures of neutrophil activation and collagen accumulation, respectively. Liver damage was significantly prevented in the bile-duct ligated rats treated with methylene blue compared with the control bile-duct ligated rats without methylene blue. Treatment with methylene blue markedly reduced activities of serum transaminase, gamma glutamine transferase and bilirubin levels as compared to bile-duct ligated rats without methylene blue. Positive immunolabelling for alpha-smooth muscle actin (alpha-SMA) was increased, especially in vascular smooth muscle cells, fibrotic septa and also around the proliferated bile ducts, after bile-duct ligation. Only weak alpha-SMA immunolabelling was seen in livers of rats treated with methylene blue. These results indicate that methylene blue can attenuate hepatic damage in extrahepatic cholestasis by reducing oxidative stress and inflammatory processes. PMID- 19217653 TI - Multiallergen immunotherapy for allergic rhinitis and asthma. AB - The English and non-English language literature on allergen immunotherapy was reviewed for studies simultaneously using 2 or more distinct allergen extracts in either subcutaneous or sublingual immunotherapy. Thirteen studies were identified, 11 using subcutaneous injections, 1 using sublingual administration, and 1 using both. In studies with adequate information, administration of 2 extracts by means of either subcutaneous immunotherapy or sublingual immunotherapy was effective. In studies using multiple allergens, 3 studies showed clear efficacy, whereas in the other 2 studies, lack of efficacy might have been due to inadequate doses of extract or omission of clinically relevant allergens in the treatment regimen. It is concluded that simultaneous administration of more than 1 allergen extract is clinically effective. However, more studies are needed, particularly with more than 2 allergen extracts and with sublingual administration. PMID- 19217651 TI - Ultrasound-enhanced thrombolysis with tPA-loaded echogenic liposomes. AB - BACKGROUND AND PURPOSE: Currently, the only FDA-approved therapy for acute ischemic stroke is the administration of recombinant tissue plasminogen activator (tPA). Echogenic liposomes (ELIP), phospholipid vesicles filled with gas and fluid, can be manufactured to incorporate tPA. Also, transcranial ultrasound enhanced thrombolysis can increase the recanalization rate in stroke patients. However, there is little data on lytic efficacy of combining ultrasound, echogenic liposomes, and tPA treatment. In this study, we measure the effects of pulsed 120-kHz ultrasound on the lytic efficacy of tPA and tPA-incorporating ELIP (t-ELIP) in an in-vitro human clot model. It is hypothesized that t-ELIP exhibits similar lytic efficacy to that of rt-PA. METHODS: Blood was drawn from 22 subjects after IRB approval. Clots were made in 20-microL pipettes, and placed in a water tank for microscopic visualization during ultrasound and drug treatment. Clots were exposed to combinations of [tPA]=3.15 microg/ml, [t-ELIP]=3.15 microg/ml, and 120-kHz ultrasound for 30 minutes at 37 degrees C in human plasma. At least 12 clots were used for each treatment. Clot lysis over time was imaged and clot diameter was measured over time, using previously developed imaging analysis algorithms. The fractional clot loss (FCL), which is the decrease in mean clot width at the end of lytic treatment, was used as a measure of lytic efficacy for the various treatment regimens. RESULTS: The fractional clot loss FCL was 31% (95% CI: 26-37%) and 71% (56-86%) for clots exposed to tPA alone or tPA with 120 kHz ultrasound. Similarly, FCL was 48% (31-64%) and 89% (76-100%) for clots exposed to t-ELIP without or with ultrasound. CONCLUSIONS: The lytic efficacy of tPA containing echogenic liposomes is comparable to that of tPA alone. The addition of 120 kHz ultrasound significantly enhanced lytic treatment efficacy for both tPA and t-ELIP. Liposomes loaded with tPA may be a useful adjunct in lytic treatment with tPA. PMID- 19217654 TI - Characteristics of childhood peanut allergy in the Australian Capital Territory, 1995 to 2007. AB - BACKGROUND: It is unknown whether clinical features of peanut allergy have changed in the past decade alongside possible increasing prevalence. OBJECTIVE: The clinical features of peanut allergy over 13 years were examined with regard to age of onset, sex distribution, severity, and incidence. METHODS: Retrospective study of 778 patients (age 4 months to 66 years) diagnosed with peanut allergy at a community-based specialist allergy practice in the Australian Capital Territory. RESULTS: Most peanut allergy (90%) developed by age 72 months. In this group, there were no significant time-dependent changes in sex distribution, reaction severity, or age of first reaction (mean/median 12/15.1 months). Later age of first reaction was associated with an increased risk of anaphylaxis in the overall population (P < .01) and in those with onset by 72 months, in whom risk increased by 22.7% (CI, 3.3-45.7) for every additional year of age (P < .02). Asthma was associated with increased risk of anaphylaxis (odds ratio, 1.9; P < .001). In children with peanut allergy, 22% experienced anaphylaxis with first exposure and 30% with anaphylaxis had preceding milder reactions. The estimated minimum incidences of peanut allergy and sensitization by age 72 months for children born in the Australian Capital Territory in 2004 were 1.15% and 1.53%, respectively (by end December 2007), compared with 0.73% and 0.84% for those born in 2001. CONCLUSION: Although most characteristics of peanut allergy have changed little over the period of the last 13 years (onset age, sex, comorbidity, severity), later onset was associated with greater risk of anaphylaxis. Our data are consistent with a rise in incidence. PMID- 19217655 TI - Integrin mediated attachment of periodontal ligament to titanium surfaces. AB - OBJECTIVE: Reducing the force between the implant and the bone by recapitulating a similar matrix has the potential to reduce implant failure. To begin to pursue the goal of creating a periodontal ligament interface between a dental implant and bone, the mechanism of cellular attachment to dental implant surfaces must be characterized. METHODS: In this study we examined the role of integrin receptors in the attachment of periodontal ligament fibroblasts to titanium surfaces utilized on dental implants; those surfaces included smooth polished titanium, acid pickled titanium, ground titanium, sandblasted and acid etched titanium, non oxidized titanium that has been sandblasted and acid etched, hydroxyapatite coated titanium, titanium plasma sprayed or uncoated titanium. For these studies integrin mediated fibroblast attachment was blocked by the integrin blocking peptide GRGDSP or anti-integrin beta1 antibody or a combination of the two. Quantitation of periodontal ligament fibroblast attachment was completed by counting cells on the various implant surfaces after culturing in vitro for 24h with and without the integrin receptor blockers. RESULTS: Antibody and peptide treatment significantly reduced the number of fibroblasts cells attached to the various implant surfaces but this effect varied significantly depending on the surface. Moreover, increased levels of peptide further decreased fibroblasts attachment in a dose dependent manner. SIGNIFICANCE: Blocking studies suggest first, that integrin receptors function in periodontal ligament attachment to titanium surfaces and second, that different integrin subunits are important in attachment to a particular surface. PMID- 19217656 TI - Novel opioid antagonists for opioid-induced bowel dysfunction and postoperative ileus. AB - Peripherally acting mu-opioid receptor antagonists methylnaltrexone and alvimopan are a new class of drugs designed to reverse opioid-induced side-effects on the gastrointestinal system without compromising pain relief. This article gives an overview of the pharmacology, the efficacy, and adverse effects of these drugs. Both compounds seem to be generally well tolerated and effective for the treatment of opioid-related bowel dysfunction and postoperative ileus. Methylnaltrexone recently received approval by the US Food and Drug Administration (FDA) and the European Medicines Agency for treatment of opioid related bowel dysfunction in patients with advanced illness. Alvimopan was recently approved by the FDA for treatment of postoperative ileus, but the use of the drug is restricted to inpatients because it has been associated with an increased rate of myocardial infarction. Further research should assess the effectiveness and safety of these drugs in clinical practice. PMID- 19217657 TI - Microscale mechanical properties of single elastic fibers: the role of fibrillin microfibrils. AB - Micromechanical properties of single elastic fibers and fibrillin-microfibrils, isolated from equine ligamentum nuchae using chemical and enzymatic methods, were determined with atomic force microscopy (AFM). Young's moduli of single elastic fibers immersed in water, devoid of or containing fibrillin-microfibrils, were determined using bending tests. Bending freely suspended elastic fibers on a micro-channeled substrate by a tip-less AFM cantilever generated a force versus displacement curve from which Young's moduli were calculated. For single elastic fibers, Young's moduli in the range of 0.3-1.5 MPa were determined, values not significantly affected by the presence of fibrillin-microfibrils. To further understand the role of fibrillin-microfibrils in vertebrate elastic fibers, layers of fibrillin-microfibrils were subjected to nano-indentation tests. From the slope of the force versus indentation curves, Young's moduli ranging between 0.56 and 0.74 MPa were calculated. The results suggest that fibrillin microfibrils are not essential for the mechanical properties of single vertebrate elastic fibers. PMID- 19217658 TI - Low molecular mass polypeptide-2 in human trophoblast: over-expression in hydatidiform moles and possible role in trophoblast cell invasion. AB - Embryo implantation involves invasion of placental extravillous trophoblast cell (EVTs) into the uterus. Hyperactive EVT invasion occurs in hydatidiform moles and choriocarcinomas. We have previously demonstrated that the 20S proteasome is involved in mouse embryo implantation and its action is mediated via regulating the expression and activities of matrix metalloproteinase (MMP)-2 and MMP-9 in the EVTs. Our objective was to investigate whether low molecular mass polypeptide 2 (LMP2), a beta subunit of the 20S proteasome, is involved in the regulation of human trophoblast invasion. Normal human placentas or placentas from hydatidiform mole patients were collected and the expression of LMP2 in different cell types including trophoblastic column (TC), cytotrophoblast cells (CTB) and syncytiotrophoblast (STB) under different pathological states were studied by immunohistochemical analysis. Furthermore, the effect of LMP2 or proteasome on cell invasion was measured by using RNAi and inhibitors in a Matrigel invasion assay system in HTR-8/SVneo cells, a human invasive extravillous trophoblast cell line. Changes in the invasion-related molecules including MMP-2 and MMP-9 were also examined by using real time PCR and gelatin zymography. We demonstrated that the expression of LMP2 in TC of partial hydatidiform mole and complete hydatidiform mole, is higher than that in TC of normal human placentas. Besides, LMP2 knockdown significantly attenuated IL-1beta-induced cell invasion in vitro, a response readily induced by proteasome inhibitors. In summary, over-expression of the 20S proteasome beta-subunit LMP2 in trophoblast cells of hydatidiform moles may contribute to its highly invasive phenotype. PMID- 19217659 TI - Blockade of central kappa-opioid receptors inhibits the antidipsogenic effect of interleukin-1beta. AB - The objective of the present study was to investigate the role of brain kappa opioid receptors (KOR) in the antidipsogenic effect promoted by third ventricle injections of interleukin-1beta (IL-1beta). Wistar male rats were submitted to three different, thirst-inducing, physiological conditions: dehydration induced by water deprivation, hyperosmolarity induced by salt-load and hypovolemia induced by polyethylene glycol subcutaneous injection. Third ventricle injections of IL-1beta significantly inhibited the increase in water intake observed in those situations. The pharmacological blockade of central KOR by the selective KOR antagonist nor-binaltorphimine (BNI) at different doses significantly inhibited the antidipsogenic effect induced by the central administration of IL 1beta in all conditions tested: dehydration, hypovolemia and hyperosmolarity. The central administration of IL-1beta failed to induce any locomotor deficit, as verified in an open field test. Stimulation of the central interleukinergic component did not result in any general suppression of ingestive behavior since no change in saccharin intake was recorded during a dessert test in animals receiving central injections of IL-1beta. Furthermore, the inhibitory effects of IL-1beta on water intake cannot be attributed to sickness-like effects induced by these compounds, since an aversion test excluded this possibility. In summary, the data shown in the present study clearly show that the antidipsogenic effect observed in rats following third ventricle injections of IL-1beta depend on the functional integrity of a brain kappa-opioid-dependent component. PMID- 19217660 TI - Assessment of environmental stress in Parablennius sanguinolentus (Pallas, 1814) of the Sicilian Ionian coast. AB - The blenny Parablennius sanguinolentus was selected as a useful bioindicator of environmental pollution. Chemical parameters in water and sediments from three different sampling sites along the Sicilian Ionian coast were determined and metal concentrations in fish muscle were measured. DNA fragmentation and oxidation in erythrocytes and hepatocytes was determined by the Comet assay and HSP70 expression levels were evaluated in the liver. The results show an increased level of chromium in sediments and high polycyclic aromatic hydrocarbon (PAH) concentrations in water at one site. The bioaccumulation of metals in muscle tissue shows high concentrations of lead in some samples. A high percentage of DNA damage in blood and liver cells, as well as high hepatic levels of HSP70, were found in all the sites. The results demonstrate the usefulness of an integrated chemical and biological approach for the determination of environmental stress. PMID- 19217661 TI - Split cord malformation incidentally discovered. PMID- 19217662 TI - Pilocytic astrocytoma arising from the pineal gland. PMID- 19217663 TI - Two cases of vertebral artery dissections with late stroke recurrences. PMID- 19217664 TI - Psychiatric disorders and personality characteristics of prisoners at regular prison wards. AB - BACKGROUND: Dutch correctional officers are trained to observe prisoners with severe mental disorder. This 'behavioural' approach is assumed to detect psychiatrically disordered prisoners with striking symptoms. On the basis of this screening procedure about 10% of the Dutch prison population is classified as needing special care or control. In the current study, what psychopathology can still be found among the remaining 90% prisoners residing at regular wards is investigated and which personality traits characterize them. When the prevalence of major mental illness would still turn out to be high in this group, the question arises whether the current, rather unstandardized, way of screening prisoners is sufficient. AIMS: To assess the prevalence of psychiatric disorders among adult Dutch prisoners on regular wards, and to determine the concurrent and convergent validity of self-report measures and their screening characteristics in such a sample. METHOD: 191 randomly selected prisoners admitted to the general wards were administered a number of tests (SCL-90, NEO-PI-R, MINI) during the first weeks of their incarceration. RESULTS: Including substance abuse, 57% of the participants suffered from one or more Axis I disorders. About seven out of ten detainees with psychopathology as assessed with the MINI did not receive professional help. The concurrent and convergent validity of the SCL-90 and the NEO-PI-R turned out to be reasonable. The predictive validity of self-report measures in detecting prisoners with an Axis I disorders or suicide risks was moderate. CONCLUSIONS: The Dutch 'behavioural approach' seems to be quite accurate in detecting prisoners with psychotic disorders. Most prisoners with other mental disorders on regular wards, however, did not receive professional help. The current study suggests that self report scales such as the SCL-90 and the NEO-PI-R may be helpful in screening detainees on important DSM-IV disorders. PMID- 19217665 TI - Arsenic geochemistry, transport mechanism in the soil-plant system, human and animal health issues. PMID- 19217666 TI - Born in 1809: Julia Margaret Cameron's photographs. PMID- 19217667 TI - Humoral immune response to abnormal MUC1 in subjects with colorectal adenoma and cancer. AB - Colorectal cancer (CRC) expresses a hypoglycosylated (abnormal) form of MUC1 different than MUC1 expressed in normal colon, which elicits antibodies in patients with CRC. This form of MUC1 is expressed in other abnormal but non malignant lesions in the colon, such as adenomatous polyps, precursors to CRC. Estimates of the prevalence of anti-MUC1 antibodies in subjects with these lesions are lacking. We evaluated IgM and IgG anti-MUC1 antibodies in 148 subjects with non-advanced adenomas (NAA), advanced adenomas (AA), colorectal cancer, hyperplastic polyps (HPP), and normal controls. We hypothesized that the prevalence of anti-MUC1 antibodies would increase along the adenoma-carcinoma sequence as more dysplastic tissues express more abnormal MUC1. Anti-MUC1 IgM was found in 5/47 (10.6%) of normals, 5/45 (11.1%) of NAA, 7/47 (14.9%) of AA, and 4/20 (20.0%) of CRC (p=0.70). The prevalence of anti-MUC1 IgG was 8/47 (17.0%) of normals, 14/45 (31.1%) of NAA, 14/47 (29.8%) of AA, and 6/20 (30.0%) of CRC (p=0.36). We found no significant differences in the prevalence of anti-MUC1 antibodies between subjects along the adenoma-carcinoma sequence. However, in an exploratory analysis, the median normalized anti-MUC1 IgG OD level of the combined abnormal groups (NAA, AA, CRC) was significantly higher than the normals (0.045 OD vs. 0.030 OD; p=0.017). Our data support further studies into the potential role of anti-MUC1 immunity in preventing progression of premalignant colon lesions to colon cancer. PMID- 19217668 TI - Clinical correlates of the worsening or emergence of suicidal ideation during SSRI treatment of depression: an examination of citalopram in the STAR*D study. AB - BACKGROUND: Untreated major depressive disorder (MDD) is a major risk factor for suicide, but some data suggest antidepressants may be associated with increased suicidal ideation (SI) in some depressed patients. The purpose of this study was to determine whether, and in whom, treatment of MDD is associated with increased or emergent SI. METHODS: Patients were treated with Citalopram, 10-60 mg/day for 12-14 weeks. A score >0 on Item 12 of the Quick Inventory of Depressive Symptomatology - Self-Report indicated the presence of SI. Worsening was defined by a >or=1 point increase. Emergent SI was defined by an increase from 0 at baseline to >or=1 during treatment. RESULTS: Of the 1909 participants with baseline SI, 57% experienced improvement in SI by their first post-baseline visit and 5% worsened. By the final visit, 74% experienced improvement and 4% worsened. Of 1721 participants without baseline SI, 7% experienced emergence by the first postbaseline visit. Of these, 63% had no SI at their final visit. Major risk factors for treatment-emergent SI at the first treatment visit were drug abuse, severe depression and melancholic features. LIMITATIONS: Main limitations are lack of a comparison group to help pinpoint whether citalopram treatment added risk or protection, a placebo group to determine whether changes in SI were related to illness factors, medication effects or other factors, and more detailed and validated measures of SI. CONCLUSIONS: SI and behaviors, core features of MDD, wax and wane in intensity before, during, and perhaps after treatment. It is clinically important to understand risk factors, maintain careful surveillance and treat as vigorously as necessary to attain remission. PMID- 19217669 TI - Patterns of early change and their relationship to outcome and follow-up among patients with major depressive disorders. AB - BACKGROUND: While highlighting the efficacy of different treatments for major depressive disorders (MDD), findings of the National Institute of Mental Health (NIMH) Treatment of Depression Collaborative Research Program (TDCRP) often are interpreted as supporting the idea of treatment non-specificity for MDD. However, heterogeneity in treatment courses and outcomes might be undetected when focusing only on types of treatment in terms of sample means without taking into account early change in treatment. METHOD: In this study, growth mixture models (GMM) were used in the completer sample of N=162 patients from the NIMH TDCRP to identify meaningful patterns of early change of depression severity that are shared by many individual patients. RESULTS: Results revealed three typical patterns of early change over the first 8 weeks of treatment irrespective of the type of treatment protocol provided: (a) moderate to severe depression with moderate early improvement, (b) moderate to severe depression with rapid early improvement, and (c) mild to moderate depression with moderate early improvement. In contrast to the type of treatment, these differential patterns of early response (together with overall pre-treatment symptom severity) predicted outcomes (i.e. depression severity) at treatment termination and over the 1.5 year follow-up period. LIMITATIONS: Due to the small sample size and the controlled setting of this study the degree to which these results can be generalized to clinical practice has to be further investigated. CONCLUSIONS: The findings provide further support for the assumption that early change is an important factor for the prediction of short- and long-term outcome in psychotherapy. PMID- 19217670 TI - Sex differences in the behavior of children with the 22q11 deletion syndrome. AB - High rates of psychiatric impairment in adults with 22q11 deletion syndrome (22q11DS, also referred to as DiGeorge or velocardiofacial syndrome) suggest that behavioral trajectories of children with 22q11DS may provide critical etiologic insights. Past findings that report Diagnostic and Statistical Manual (DSM) diagnoses are extremely variable; moreover, sex differences in behavior have not yet been examined. Child Behavior Checklist (CBCL) ratings from 82 children, including 51 with the 22q11DS and 31 control siblings, were analyzed. Strikingly consistent with rates of psychiatric impairment among affected adults, 25% of children with 22q11DS had high CBCL scores for Total Impairment, and 20% had high CBCL Internalizing Scale scores. Males accounted for 90% of high Internalizing scores and 67% of high Total Impairment scores. Attention and Social Problems were ubiquitous; more affected males than females (23% vs. 4%) scored high on Thought Problems. With regard to CBCL/DSM overlap, 20% of affected males as compared with 0 affected females had one or more high CBCL ratings in the absence of a DSM diagnosis. Behaviors of children with 22q11DS are characterized by marked sex differences when rated dimensionally, with significantly more males experiencing Internalizing and Thought Problems. Categorical diagnoses do not reflect behavioral differences between male and female children with 22q11DS, and may miss significant behavior problems in 20% of affected males. PMID- 19217671 TI - Therapeutic activities of intravenous immunoglobulins in multiple sclerosis involve modulation of chemokine expression. AB - The objective of this study was to identify genes that are differentially expressed in peripheral T cells of patients with MS exacerbation receiving treatment with IVIG. Using microarray analysis, we identified 360 genes that were at least two-fold up- or down-regulated. The expression of four representative genes (PTGER4, CXCL5, IL11 and CASP2) was confirmed by quantitative PCR. Four of the differentially expressed genes encode chemokines (CXCL3, CXCL5, CCL13 and XCL2) that are involved in directing leukocyte migration. We suggest that the modulation of chemokine expression in peripheral T cells contributes to the beneficial activity of IVIG in patients with MS exacerbation. PMID- 19217672 TI - Associations of personal and mobility resources with subjective well-being among older adults in Italy and Germany. AB - The purpose of this study was to investigate the patterns of associations of personal and mobility resources with positive well-being in two sample populations of older adults from Germany (n=1518) and Italy (n=600). A correlation study showed which variables correlated the closest with the well being components, then four logistic regression analyses were performed, one for each well-being component such as the positive and negative affect schedule (PANAS) positive scale, mobility satisfaction, health satisfaction and life satisfaction. High positive scores of PANAS and satisfaction scales were found both for Germany and Italy. As expected and in accordance with previous studies, components utilized as a measure of subjective well-being correlated significantly with each other in both countries, confirming their validity as instruments of evaluation for several different life domains. A considerable result of this study was that the driving car is an important predictor of well being in both countries. The findings underlined a variety of associations among the mobility resources and the subjective perception of well-being. Our work supports the importance of being aware of the main variables influencing the positive dimension of well-being for creating a better understanding the needs and goals of older people. Moreover, the present study shows that individual environment interaction should be considered in evaluating personal well-being, especially in older people. PMID- 19217673 TI - Statin therapy in the elderly: a review. AB - Cardiovascular morbidity is the leading cause of mortality in the developed nations. Elevated serum cholesterol is a major risk factor for ischemic heart disease, one of the common cardiovascular morbidity in older adults, statins have been shown to be effective in reducing serum cholesterol and improving outcomes. Hypercholesterolemia is common in older adults and is one of the major modifiable risk factors. Yet, these patients have often been excluded from major clinical trials of statins and evidence suggests of their underuse. Data from recent clinical trials of statins indicate that the elderly patients with the highest cardiovascular risk are likely to derive the most benefits from cholesterol lowering. With the aging of the population, the prevalence of hypercholesterolemia and cardiovascular morbidity is likely to increase. In this review we evaluate the evidence for the use of statins in older adults. PMID- 19217674 TI - Prevalence and related influencing factors of depressive symptoms for empty-nest elderly living in the rural area of YongZhou, China. AB - The number of empty-nest elderly is on the rise, especially in rural areas of China. The empty-nest elderly were concerned from social and demographic perspective, but researches on mental health issues of the aged were still rare in China. To clarify the prevalence of depression among empty-nest elderly in a rural area of YongZhou, Hunan and evaluate the impact of social support, coping style and socio-demographic factors on depression of the empty-nest elderly, with the goal of decreasing the rate of depression in empty-nest elderly, a questionnaire survey was conducted on 415 elderly persons. The determined prevalence of depressive symptoms was 74.46%. The empty-nest group, in comparison with the non-empty nest group, had higher levels of depression. Multivariate linear regression analysis showed associations between depressive symptoms and negative coping style, support utilization, religious beliefs, economic status, marital status and subjective support. The results suggested that the strategy to alleviate the depression in empty-nest elderly can be considered by the society, village committee, the family members and the empty-nest elderly subjects themselves, which could control effectively the incidence of depression. PMID- 19217675 TI - Exercise capacity at submaximal heart rate and prognosis. AB - INTRODUCTION: Submaximal exercise testing can be useful for individuals with limitations to performing maximal exercise. Recent studies suggested that a low workload at a heart rate 100 beats/minute (HR(100)) was associated with a greater risk of cardiovascular (CV) mortality than maximal exercise capacity. This study evaluated the prognostic value of METs achieved at HR(100) (METs(100)) in patients referred for treadmill testing. METHOD: We studied 1446 patients (56+/ 12 years; 76 females) without heart failure or beta-blockers treatment (1997 2004). RESULTS: During a period of 7.0+/-2.3 years, 35 (2.5%) patients died from CV causes. Compared to survivors, the non-survivors were older (69+/-9 vs. 56+/ 12 years, p<0.001); had a higher prevalence of diabetes (27% vs. 14%, p=0.04), coronary artery disease (57% vs. 25%, p<0.05) and stroke (9% vs. 2%, p<0.001). Non-survivors had lower Duke Treadmill Scores (DTS) (2.8+/-6.8 vs. 9.7+/-5.5; p<0.001) and exercise capacity (7.5+/-3.3 vs.11.0+/-3.8 METs, p<0.001). At HR(100), METs (median (range): 3.8 (2.8-4.0) vs. 3.5 (3.3-3.5)) and %HR reserve achieved (45+/-13% vs. 34+/-17%; p<0.001) were higher in non-survivors. In Cox model, age-adjusted METs(100) was not a significant predictor of CV mortality. In contrast, each one MET increase in exercise capacity was associated with a 17% increase in survival (HR=0.83, 95% CI 0.73-0.93, p=0.002). DTS was also a significant predictor of CV mortality. CONCLUSION: In our population, METs at HR(100) was not a significant predictor of CV mortality. PMID- 19217676 TI - Exercise training in adults with congenital heart disease: feasibility and benefits. AB - BACKGROUND: Regular physical activity is associated with a range of health benefits; however the influence of regular exercise training on exercise capacity and quality of life in adults with congenital heart disease [ACHD] has not been previously reported. METHODS: Prospective study of patients attending the ACHD clinic in a supra-regional centre, assessing feasibility and effects of exercise training. There were 3 phases: Phase I: Initial assessment including quality of life and physical activity questionnaires; treadmill exercise test; physical activity assessment with accelerometers [Caltrac and Actigraph]; Phase II: Exercise training (home-based walking 5/7 days) for 10 weeks; Phase III: Re assessment. RESULTS: Sixty-one adults [36 males; mean age 31.7+/-10.9 yrs] were divided into 3 groups according to NYHA class. Fifty patients completed the intervention and all pre-post assessments. Group I (n=21; 13 males), Group II (n=16; 10 males), Group III (n=13, 10 males). Median body mass index was 23.8; 12 were overweight (BMI 25-29.9) and 7 obese (BMI>30). The scores of the quality of life and physical activity questionnaires increased significantly after exercise training in all three groups. Similar increases were seen in the treadmill test duration and in physical activity levels assessed by the Caltrac and the Actigraph in all groups. CONCLUSIONS: A simple physical activity intervention like regular walking is feasible, safe and significantly increases the exercise capacity of adult patients at all stages of congenital heart disease. It is also helpful in improving the quality of life by improving physical self-perception, satisfaction with life, physical activity levels and general health. PMID- 19217677 TI - Poisson sampling: a sampling strategy for concurrently establishing freedom from disease and estimating population characteristics. AB - Surveys of animal populations are often designed to either demonstrate freedom from disease or to estimate parameters that describe the population, such as disease prevalence, proportion of vaccinated animals, or average animal weight and value. Targeted surveillance is a sampling approach where animals are selected for testing based on the presence of characteristics that indicate a higher probability of disease. This approach can substantially reduce the sample size that is required to demonstrate freedom from disease, but inferences about other population parameters are generally not possible because the sample design often lacks the properties required for making inferences in a traditional survey sample. Determining which animals to sample can also be difficult when either more than one characteristic exists or the characteristic is a continuous attribute, such as age or weight. Poisson sampling is an unequal probability sampling design that can provide efficiencies similar to targeted surveillance while allowing inferences for other population parameters. The adaptation of Poisson sampling to animal surveys is described. A simulation study, based on sampling a flock of sheep, is used to demonstrate the reductions in sample size that are possible with Poisson sampling. The study showed that the sample size required for a flock-level sensitivity of 0.95 when using Poisson sampling was less than half that required when using simple random sampling. The performance of estimators for prevalence of scrapie and distribution of genotypes are also compared. PMID- 19217678 TI - The impact of poor health on academic performance: New evidence using genetic markers. AB - This paper examines the influence of health conditions on academic performance during adolescence. To account for the endogeneity of health outcomes and their interactions with risky behaviors we exploit natural variation within a set of genetic markers across individuals. We present evidence that specific genetic markers have good statistical properties to identify the impacts of ADHD, depression and obesity. These markers help reveal a new dynamism from poor health to lower academic achievement with substantial heterogeneity in their impacts across genders. Our investigation further exposes the considerable challenges in identifying health impacts due to the prevalence of comorbid health conditions, with clear implications for the health economics literature. PMID- 19217679 TI - Efficacy of myrtle oil against Salmonella Typhimurium on fresh produce. AB - The antimicrobial activity of myrtle leaves (Myrtus communis) oil was tested against the nalidixic acid resistant strain of Salmonella Typhimurium ATCC 13311. An inoculum (100 microl, ca.10(8) cfu/ml) was deposited on the skin of whole tomatoes and 10 g of shredded iceberg lettuce, dried for 2 h at 22 degrees C and held for 22 h at 4 degrees C before treatments. Inoculated iceberg lettuce (3.51 3.99 log cfu/g) and tomatoes (3.47-4.86 log cfu/tomato) were treated with three different washing procedures for 5, 10, 15 and 20 min; washing with sterile distilled water (control), washing with three different concentrations of myrtle leaves oil and the last treatment was a combination of washing with myrtle leaves oil and then rinsing in sterile distilled water for 1 min. Washing with myrtle leaves oil with or without rinsing procedures caused significant reduction in S. Typhimurium population compared with the control after treatment for four different times (p<0.05). There is no significant difference between washing times in reduction of S. Typhimurium (p>0.05). The maximum logarithmic reductions of 1.66 cfu/g-1.89 cfu/tomato were respectively obtained on iceberg lettuce and tomatoes treated with 1000 ppm myrtle leaves oil without any rinsing treatment. The results suggest that the use of myrtle leaves oil is an innovative and useful tool as an alternative to the use of chlorine or other synthetic disinfectants in fruits and vegetables, especially for organic products. PMID- 19217680 TI - Genetic manipulation of HSP26 and YHR087W stress genes may improve fermentative behaviour in wine yeasts under vinification conditions. AB - Throughout wine production yeast cells are affected by a plethora of stress conditions that compromise their ability to carry out the whole process. In recent years important knowledge about the mechanisms involved in stress response in both laboratory and wine yeast strains has been obtained. Several studies have indicated that a correlation exists between stress resistance, expression of stress response genes and fermentative behaviour. In this work we introduce several genetic manipulations in two genes induced by several stress conditions: HSP26 (which encodes a heat shock protein) and YHR087W (encoding a protein of unknown function) in two different wine yeasts, ICV16 and ICV27. These manipulations include expression in multicopy and centromeric plasmids, and substitution of the promoter in one of the genomic copies of these genes for that of the SPI1 gene, encoding for a cell wall protein of unknown function, or the PGK1 gene, which encodes the phosphoglycerate kinase glycolytic enzyme. Our results indicate that some of these modifications result in strains with higher expression of these genes, better resistance to certain stress conditions, and even improved fermentative behaviour. The modifications of the YHR087W gene are particularly interesting, and suggest an important role of this gene in the vinification process. PMID- 19217681 TI - Reduction of Vibrio vulnificus in pure culture, half shell and whole shell oysters (Crassostrea virginica) by X-ray. AB - The purpose of this investigation was to study the inactivation effect of X-ray treatments on inoculated Vibrio vulnificus in pure culture, half shell and whole shell oysters to achieve a 5.0 log reduction, which is recommended by the Interstate Shellfish Sanitation Conference and the Food and Drug Administration. A mixed culture of three V. vulnificus strains was used to prepare the pure culture and inoculated oysters. The pure culture and inoculated oysters were treated with 0.0, 0.1, 0.5, 0.75, 1.0, 1.5, 2.0, and 3.0 kGy X-ray at 22 degrees C and 50-60% relative humidity. Surviving bacterial populations in the pure culture and inoculated oysters were enumerated using overlay-plating method [with a non-selective media (trypticase soy agar) followed by a selective medium (Modified Cellobiose-Polymyxin B-Colistin) and most probable number (MPN) method. Greater than a 6-log reduction of V. vulnificus was achieved with 0.75, 1.0 3.0 kGy X-ray in pure culture, half shell and whole shell oysters, respectively. Treatment with 0.75 kGy X-ray significantly (p<0.05) reduced the inherent microorganisms in half shell oysters, to less than the detectable limit (<1 log CFU/g). The maximum dose (3.0 kGy) of X-ray treatment did not affect the survivability of live oysters. PMID- 19217682 TI - Impact of coffee consumption on the gut microbiota: a human volunteer study. AB - The impact of a moderate consumption of an instant coffee on the general composition of the human intestinal bacterial population was assessed in this study. Sixteen (16) healthy adult volunteers consumed a daily dose of 3 cups of coffee during 3 weeks. Faecal samples were collected before and after the consumption of coffee, and the impact of the ingestion of the product on the intestinal bacteria as well as the quantification of specific bacterial groups was assessed using nucleic acid-based methods. Although faecal profiles of the dominant microbiota were not significantly affected after the consumption of the coffee (Dice's similarity index=92%, n=16), the population of Bifidobacterium spp. increased after the 3-week test period (P=0.02). Moreover, in some subjects, there was a specific increase in the metabolic activity of Bifidobacterium spp. Our results show that the consumption of the coffee preparation resulting from water co-extraction of green and roasted coffee beans produce an increase in the metabolic activity and/or numbers of the Bifidobacterium spp. population, a bacterial group of reputed beneficial effects, without major impact on the dominant microbiota. PMID- 19217683 TI - Consensus on the leadership of hospital CEOs and its impact on the participation of physicians in improvement projects. AB - OBJECTIVES: The success of a Dutch program to disseminate quality improvement projects depends on the participation of physicians working in program hospitals. The leadership of hospital executives (CEOs) is considered an important explanation. This study aims to determine whether the relation, between the extent to which physicians notice their CEOs stimulate improvement initiatives and the number of projects joined by physicians, is moderated by the consensus among physicians working in the same hospital. METHODS: Multilevel analyses are applied on data of 286 physicians from eight hospitals to: (1) estimate whether participation depends on noticing if CEOs stimulate improvement, (2) test if an individual's participation differs when more colleagues have the same opinion (effect modification). RESULTS: Significant moderator effects are found. The participation of physicians, noticing that CEOs stimulate improvement is higher when more colleagues share this opinion. For physicians not knowing whether improvement is encouraged, higher consensus coincides with lower participation. CONCLUSION: Project involvement of physicians depends on their consensus about encouragement by CEOs. This confirms the importance of strategic leaders in dissemination programs. Further research is recommended into causes of CEO leadership visibility and methods to strengthen leadership climate. PMID- 19217684 TI - GWA studies: rewriting the story of IBD. AB - Genome-wide association (GWA) studies are substantially improving our understanding of the molecular pathways leading to inflammatory bowel diseases (IBD). This is a result of the nature of these studies, which are comprehensive - leading to a dramatic increase in the number of validated genetic risk factors - and unbiased - leading to the identification of novel pathways not previously suspected in IBD. Such discoveries are not only driving the functional studies to understand the mechanisms by which genetic variants modify an individual's susceptibility to disease, but also hold the promise of guiding the development of more effective treatment strategies. In this review, we discuss how GWA studies are enabling us to rewrite the story of IBD pathogenesis, focusing on the interleukin-23 and autophagy pathways. PMID- 19217685 TI - An enhanced algorithm for knee joint sound classification using feature extraction based on time-frequency analysis. AB - Vibroarthrographic (VAG) signals, generated by human knee movement, are non stationary and multi-component in nature and their time-frequency distribution (TFD) provides a powerful means to analyze such signals. The objective of this paper is to improve the classification accuracy of the features, obtained from the TFD of normal and abnormal VAG signals, using segmentation by the dynamic time warping (DTW) and denoising algorithm by the singular value decomposition (SVD). VAG and knee angle signals, recorded simultaneously during one flexion and one extension of the knee, were segmented and normalized at 0.5 Hz by the DTW method. Also, the noise within the TFD of the segmented VAG signals was reduced by the SVD algorithm, and a back-propagation neural network (BPNN) was used to classify the normal and abnormal VAG signals. The characteristic parameters of VAG signals consist of the energy, energy spread, frequency and frequency spread parameter extracted by the TFD. A total of 1408 segments (normal 1031, abnormal 377) were used for training and evaluating the BPNN. As a result, the average classification accuracy was 91.4 (standard deviation +/-1.7) %. The proposed method showed good potential for the non-invasive diagnosis and monitoring of joint disorders such as osteoarthritis. PMID- 19217686 TI - Fuzzy-stochastic characterization of site uncertainty and variability in groundwater flow and contaminant transport through a heterogeneous aquifer. AB - Site variabilities and uncertainties in data and information lead to significant spread in results of groundwater flow and contaminant transport models. A framework for hybrid propagation of random uncertainties represented by probability theory; nonrandom uncertainties represented by fuzzy set theory; and site variabilities represented by geostatistics was developed in this research. A case study was provided to explain the computational algorithm. The methodology presented here can be applied to complex environments where there are site variabilities as well as uncertainties of different kinds. The algorithm is suited when uncertainties in some variables may be best represented as fuzzy numbers whereas in others as probability distributions and both form part of the same governing equation. PMID- 19217687 TI - Stem juice production of the C4 sugarcane (Saccharum officinarum) is enhanced by growth at double-ambient CO2 and high temperature. AB - Two cultivars of sugarcane (Saccharum officinarum cv. CP73-1547 and CP88-1508) were grown for 3 months in paired-companion, temperature-gradient, sunlit greenhouses under daytime [CO2] of 360 (ambient) and 720 (double ambient) micromol mol(-1) and at temperatures of 1.5 degrees C (near ambient) and 6.0 degrees C higher than outside ambient temperature. Leaf area and biomass, stem biomass and juice and CO2 exchange rate (CER) and activities of ribulose bisphosphate carboxylase-oxygenase (Rubisco) and phosphoenolpyruvate carboxylase (PEPC) of fully developed leaves were measured at harvest. On a main stem basis, leaf area, leaf dry weight, stem dry weight and stem juice volume were increased by growth at doubled [CO2] or high temperature. Such increases were even greater under combination of doubled [CO2]/high temperature. Plants grown at doubled [CO2]/high temperature combination averaged 50%, 26%, 84% and 124% greater in leaf area, leaf dry weight, stem dry weight and stem juice volume, respectively, compared with plants grown at ambient [CO2]/near-ambient temperature combination. In addition, plants grown at doubled [CO2]/high temperature combination were 2-3 fold higher in stem soluble solids than those at ambient [CO2]/near-ambient temperature combination. Although midday CER of fully developed leaves was not affected by doubled [CO2] or high temperature, plants grown at doubled [CO2] were 41-43% less in leaf stomatal conductance and 69-79% greater in leaf water-use efficiency, compared with plants grown at ambient [CO2]. Activity of PEPC was down-regulated 23-32% at doubled [CO2], while high temperature did not have a significant impact on this enzyme. Activity of Rubisco was not affected by growth at doubled [CO2], but was reduced 15-28% at high temperature. The increases in stem juice production and stem juice soluble solids concentration for sugarcane grown at doubled [CO2] or high temperature, or at doubled [CO2]/high temperature combination, were partially the outcome of an increase in whole plant leaf area. Such increase would enhance the ongoing and cumulative photosynthetic capability of the whole plant. The results indicate that a doubling of [CO2] would benefit sugarcane production more than the anticipated 10-15% increase for a C4 species. PMID- 19217688 TI - Benzyladenine metabolism and temporal competence of Pinus pinea cotyledons to form buds in vitro. AB - Germination negatively affects adventitious shoot formation induced by cytokinins in pine cotyledons. To investigate the causes of this decrease in the organogenic response, uptake and metabolism of benzyladenine (BA) were studied in stone pine cotyledons (Pinus pinea) isolated from in vitro germinating embryos and cultured in bud induction medium. As embryos grew, cotyledons showed a progressive decrease in the amount of BA taken up from the medium. BA was barely metabolized; however, a BA metabolite previously undescribed in conifers was found. It was identified as a glucoside of the BA riboside, a type of metabolite recently described in other gymnosperms. Data revealed that differences in the organogenic capacity of P. pinea cotyledons associated with embryo germination are related primarily to their ability to absorb BA from the bud induction medium. PMID- 19217689 TI - Accuracy of teledermatology for nonpigmented neoplasms. AB - BACKGROUND: Studies of teledermatology utilizing the standard reference of histopathology are lacking. OBJECTIVE: To compare accuracy of store-and-forward teledermatology for non-pigmented neoplasms with in-person dermatology. METHODS: This study was a repeated-measures equivalence trial involving veterans with non pigmented skin neoplasms. Each lesion was evaluated by an in-person dermatologist and a teledermatologist; both generated a primary diagnosis, up to two differential diagnoses, and management plan. The primary outcome was aggregated diagnostic accuracy (percent correct matches of any chosen diagnosis with histopathology). Secondary outcomes included management plan accuracy (percent correct matches with expert panel management plan). Additional analyses included evaluation of the incremental effect of using polarized light dermatoscopy in addition to standard macro images, and evaluating benign and malignant lesion subgroups separately. RESULTS: Most of the 728 participants were male (97.8%) and Caucasian (98.9%). The aggregated diagnostic accuracy (primary outcome) of teledermatology (macro images) was not equivalent (95% confidence interval [CI] for difference within +/-10%) and was inferior (95% CI lower bound <10%) to in person dermatology for all lesions and the subgroups of benign and malignant lesions. However, management plan accuracy was equivalent. Teledermatology aggregated diagnostic accuracy using polarized light dermatoscopy was significantly better than for macro images alone (P = .0017). The addition of polarized light dermatoscopy showed the same pattern for malignant lesions, but not for benign lesions. Most interestingly, for malignant lesions, the addition of polarized light dermatoscopy yielded equivalent aggregated diagnostic accuracy rates. LIMITATIONS: Non-diverse study population. CONCLUSIONS: Using macro images, the diagnostic accuracy of teledermatology was inferior to in-person dermatology, but accuracy of management plans was equivalent. The addition of polarized light dermatoscopy yielded significantly better aggregated diagnostic accuracy, but management plan accuracy was not significantly improved. For the important subgroup of malignant lesions, the addition of polarized light dermatoscopy yielded equivalent diagnostic accuracy between teledermatologists and clinic dermatologists. PMID- 19217690 TI - Seasonal variation of Stevens-Johnson syndrome and toxic epidermal necrolysis associated with trimethoprim-sulfamethoxazole. AB - BACKGROUND: Stevens-Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN) are rare and severe cutaneous adverse reactions to medications and infections. OBJECTIVE: We sought to determine whether a seasonal variation to SJS and TEN exists and to define the characteristics in our tertiary referral hospital. METHODS: A retrospective chart review of 50 patients from 1995 through 2007 was performed and statistically analyzed. RESULTS: The most common medication implicated as a cause of SJS/TEN was trimethoprim-sulfamethoxazole (TMX) (26%). A seasonal trend, favoring springtime, was observed for the total number of cases of SJS and TEN (P = .34). There was a significant increase in cases due to TMX (53%) occurring in spring compared to other seasons (P = .002). These patients were significantly younger (37.8 +/- 13.7) than other patients with SJS and TEN (53.7 +/- 16.4) (P = .003). Their overall mortality (1 death) and average SCORTEN value (1.62 +/- 1.6) was also significantly lower (P = .04 and 0.03, respectively). Based on outpatient pharmacy records, there was no increase in TMX prescriptions filled during the spring. LIMITATIONS: The study was limited by reliance on chart data, the use of inpatient records, and number of patients. CONCLUSIONS: A seasonal variation in SJS and TEN caused by TMX affecting younger patients may exist. PMID- 19217691 TI - Comparison of a 585-nm pulsed dye laser and a 1064-nm Nd:YAG laser for the treatment of acne scars: A randomized split-face clinical study. AB - BACKGROUND: No studies have reported a comparison of the pulsed dye laser (PDL) and the 1064-nm long-pulsed neodymium:yttrium-aluminum-garnet (Nd:YAG) laser treatment of acne scars in the same patient. OBJECTIVE: To compare the efficacies of these two lasers in the treatment of acne scars. METHODS: Eighteen patients received 4 sessions of PDL or Nd:YAG laser at 2-week intervals in a randomized split-face manner. RESULTS: Both lasers induced notable and comparable improvement in the appearance of acne scars, particularly superficial scars, with significant reductions in the scores associated with the clinical evaluation scale for acne scarring (ECCA). Histologic evaluations revealed significant increases in collagen production and deposition following both lasers. Patient satisfaction scores concurred with these improvements. Ice-pick scars and boxcar scars tended to respond better to PDL and Nd:YAG lasers, respectively. LIMITATIONS: The number of subjects was small. CONCLUSIONS: Both lasers are effective modalities for the treatment of acne scars. Optimal outcomes might be achieved considering scar types and responses to a specific laser. PMID- 19217692 TI - Thrombocytopenia associated with the use of anti-tumor necrosis factor-alpha agents for psoriasis. AB - BACKGROUND: Thrombocytopenia has been reported to be associated with efalizumab therapy, but has only sporadically been reported with other anti-tumor necrosis factor alfa (TNF-alpha) agents. OBJECTIVE: To describe the frequency of thrombocytopenia in a cohort of patients who underwent biological therapies for psoriasis. METHODS: This was a retrospective observational study of 93 patients. RESULTS: One hundred eighteen courses of biological therapies were administered to 93 patients. Four of 67 patients who received anti-TNF-alpha agents developed drug-induced thrombocytopenia during treatment, compared with none of the 51 patients receiving efalizumab therapy. The platelet count recovered after suspension of anti-TNF-alpha agents in 3 patients and relapsed after re-exposure in two patients. The overall estimated frequency of thrombocytopenia in our cohort was 4.30% (95% confidence interval [CI], 0% to 6.2%). LIMITATIONS: These findings should be validated in larger studies. CONCLUSIONS: Drug-induced thrombocytopenia is a potential side effect of anti-TNF-alpha agents. Immediate monitoring of platelet counts is recommended if autoimmunity is suspected. PMID- 19217693 TI - Persistent hypoglycemia in a patient with diabetes taking etanercept for the treatment of psoriasis. AB - We report a patient with type 2 diabetes mellitus who, while treated with the antitumor necrosis factor-alpha blocking agent etanercept for severe plaque psoriasis, experienced persistent hypoglycemia requiring the lowering and eventual elimination of his previous insulin regimen. After 20 months of therapy on etanercept, his plaque psoriasis markedly improved, whereas both his fasting blood sugars and glycosylated hemoglobin A(1c) decreased. Hypoglycemia can be a serious side effect of etanercept in patients already on antidiabetic medications known to cause hypoglycemia, such as sulfonylureas, meglitinides, and insulin. Thus, it is important for dermatologists treating patients with diabetes and antitumor necrosis factor-alpha agents for psoriasis to be aware of potential hypoglycemia and to adjust antidiabetes therapy accordingly. PMID- 19217695 TI - A randomized, crossover comparison of injected buffered lidocaine, lidocaine cream, and no analgesia for peripheral intravenous cannula insertion. AB - STUDY OBJECTIVE: We compare pain and anxiety associated with peripheral intravenous (IV) cannula insertion after pretreatment with no local anesthesia, 4% lidocaine cream, or subcutaneously injected, buffered 1% lidocaine. METHODS: In a randomized, crossover design, 3 peripheral IVs were inserted in each of 70 medical students or nurses. In random order, insertion sites were pretreated with nothing, lidocaine cream, or injected, buffered lidocaine. After each IV insertion, subjects recorded pain, anxiety, and preference (as patient and provider) for each technique on a 10-point numeric rating scale. Higher scores indicated greater pain, anxiety, and preference. RESULTS: Median pain scores (interquartile range [IQR]) were 7 (4 to 8) without local anesthesia, 3 (2 to 5) with lidocaine cream, and 1 (1 to 2) with injected, buffered lidocaine. Median anxiety scores (IQR) were 4 (2 to 7) without local anesthesia, 2 (1 to 4) with lidocaine cream, and 2 (1 to 3) with injected, buffered lidocaine. There was no detectable difference in anxiety scores between lidocaine cream and injected, buffered lidocaine. Most IV placement attempts were successful, regardless of technique. Seventy percent of subjects indicated they would "always" request buffered lidocaine for peripheral IV insertion. CONCLUSION: In adult health care providers, pain and anxiety associated with peripheral IV insertion is significantly reduced by using topical lidocaine cream or injected, buffered lidocaine. Injected, buffered lidocaine reduces IV insertion pain more than lidocaine cream, without affecting success. Adults desire the use of local anesthetic techniques for IV insertion for themselves and for their patients. PMID- 19217694 TI - Guidelines of care for the management of psoriasis and psoriatic arthritis. Section 3. Guidelines of care for the management and treatment of psoriasis with topical therapies. AB - Psoriasis is a common, chronic, inflammatory, multi-system disease with predominantly skin and joint manifestations affecting approximately 2% of the population. In this third of 6 sections of the guidelines of care for psoriasis, we discuss the use of topical medications for the treatment of psoriasis. The majority of patients with psoriasis have limited disease (<5% body surface area involvement) and can be treated with topical agents, which generally provide a high efficacy-to-safety ratio. Topical agents may also be used adjunctively for patients with more extensive psoriasis undergoing therapy with either ultraviolet light, systemic or biologic medications. However, the use of topical agents as monotherapy in the setting of extensive disease or in the setting of limited, but recalcitrant, disease is not routinely recommended. Treatment should be tailored to meet individual patients' needs. We will discuss the efficacy and safety of as well as offer recommendations for the use of topical corticosteroids, vitamin D analogues, tazarotene, tacrolimus, pimecrolimus, emollients, salicylic acid, anthralin, coal tar, as well as combination therapy. PMID- 19217696 TI - The use of penicillin skin testing to assess the prevalence of penicillin allergy in an emergency department setting. AB - STUDY OBJECTIVE: Patient-reported penicillin allergies are often unreliable and can result in unnecessary changes in antibiotic therapy. Although penicillin allergy skin testing is commonly performed in allergy clinics, it has not been used in emergency departments (EDs) to verify self-reported allergies. We hypothesize that ED-based testing is possible and that the false-positive rate of patients with self-reported penicillin allergy are greater than 90%. METHODS: This prospective observational cohort study enrolled a convenience sample of ED patients with a self-reported penicillin allergy. Patients were enrolled by one of 2 emergency physicians who performed skin prick and intracutaneous tests with penicillin major and minor determinants. The total testing time was 30 minutes. The proportion of false-positive self-reported allergies was computed with 95% confidence intervals (CIs) by using the score method. RESULTS: A total of 150 patients (mean age 42 years; SD 16 years; 46% men; 47% black) were enrolled. The false-positive rate for self-reported penicillin allergy was 137 of 150 (91.3%; 95% CI 85.3% to 95.1%). There were no adverse reactions associated with penicillin skin testing. Compared with patients with a false-positive penicillin allergy result (confirmed by negative penicillin skin testing result), patients reporting a true penicillin allergy confirmed by positive penicillin skin test results tended to be more frequently men (61.5% versus 44.5%; Delta 17.0%; 95% CI -13.5% to 42%), black (69.2% versus 44.5%; Delta 24.7%; 95% CI -6.9% to 46.8%), and have no family history of drug allergy (7.7% versus 17.5%; Delta9.8%; 95% CI 20.9% to 20.4%), but self-reported other drug allergies more frequently (61.5% versus 38.7%; Delta 22.9%; 95% CI -7.7% to 47.5%). CONCLUSION: Penicillin skin testing is feasible in the ED setting. A substantial number of patients who self report a penicillin allergy do not exhibit immunoglobulin E-mediated sensitization to penicillin major and minor determinants. Penicillin testing in the ED may allow the use of more appropriate antibiotics for patients presenting with a history of penicillin allergy. PMID- 19217697 TI - Synthesis and antitumor activity of some new xanthotoxin derivatives. AB - The condensation of 4-amino-9-methoxy psoralene (4-aminoxanthotoxin) with some aromatic aldehydes led to the formation of 4-arylimine xanthotoxin derivatives 2a h, which were cyclized with mercaptoacetic acid to afford the thiazolidinone derivatives 3a-h. On the other hand, the reaction of aminoxanthotoxin 1 with some anhydrides afforded 4-imidione derivatives 3a-d. When 1 reacted with some isothiocyanates, the thiourea derivatives 5a-c were obtained but the thiourea derivative 6 was obtained when 1 reacted with ammonium thiocyanate. The thiourea derivative 6 was cyclized by the reaction with monochloroacetic acid in the presence of sodium acetate to give aminothiazolidinone derivative 7, but when the same reaction is carried out in the presence of pyridine, the thioxoimidazolidinone 8 was formed. The condensation of xanthotoxin sulphonamide with aromatic aldehydes gave the aryliminosulphonyl derivatives 9a-e. Xanthotoxin sulphonyl hydrazine condensed with some anhydride afforded sulphonic acid imide derivatives 10a-c. The antitumor and cytotoxic activities of 9 synthesized derivatives were tested, five compounds were found to be active, they inhibited the growth of HeLa cells. PMID- 19217698 TI - Synthesis and antileishmanial activity of novel 2,4,6-trisubstituted pyrimidines and 1,3,5-triazines. AB - A series of 2,4,6-trisubstituted pyrimidines and 1,3,5-triazines have been synthesized and screened for their in vitro and in vivo antileishmanial activity against Leishmania donovani. Among all, 14 compounds have shown promising inhibition of 80-100% at 10 microg/ml against promastigotes and IC(50) in the range of 0.89-9.68 microg/ml against amastigotes. Three compounds 13, 32 and 33 with good selectivity index (S.I.) were screened for their in vivo activity in golden hamsters (Mesocricetus auratus) infected with MHOM/IN/80/Dd(8) strain of L. donovani and have shown moderate in vivo inhibition of 48-56% at a dose of 50 mg/kg x 5, i.p. route for 5 days. PMID- 19217699 TI - Synthesis and biological evaluation of abietic acid derivatives. AB - A series of C18-oxygenated derivatives of abietic acid were synthesized and evaluated for their cytotoxic, antimycotic, and antiviral activities. In general, the introduction of an aldehyde group at C18 did improve the resultant bioactivity, while the presence of an acid or alcohol led to less active compounds. PMID- 19217700 TI - Re: Letter to the editor: will any wire do? PMID- 19217701 TI - Heavy metal contamination in surface sediments of Yangtze River intertidal zone: an assessment from different indexes. AB - Surface sediments (0-5 cm) from 59 stations within the Yangtze River intertidal zone (YRIZ) were sampled for metal contamination analysis in April and August 2005. The concentrations ranged (in mg kg(-1) dry weight): Al, 40,803-97,213; Fe, 20,538-49,627; Cd, 0.12-0.75; Cr, 36.9-173; Cu, 6.87-49.7; Mn, 413-1,112; Ni, 17.6-48.0; Pb, 18.3-44.1; and Zn, 47.6-154; respectively. Among the 59 sampling stations, enrichment factors (EF) indicate enrichment of Cd (52 stations), Cr (54 stations), Cu (5 stations), Ni (26 stations), Pb (5 stations) and Zn (5 stations). Geoaccumulation indexes (I(geo)) also suggest individual metal contamination in localized areas. This study indicates that Cd, Cr and Ni enrichment in the YRIZ sediment is widespread whereas Cu, Mn, Pb and Zn enrichment is localized or nonexistent. Factor and cluster analyses indicate that Cd is associated with total organic carbon whereas Cu, Cr, Ni, Pb and Zn have a close association with Mn. PMID- 19217703 TI - Idiographic assessment: conceptual and psychometric foundations of individualized behavioral assessment. AB - Idiographic assessment is the measurement of variables and functional relations that have been individually selected, or derived from assessment stimuli or contexts that have been individually tailored, to maximize their relevance for the particular individual. This article first reviews various definitions and clinical applications of idiographic assessment. Several properties of behavior problems and causal relations provide the conceptual basis for idiographic assessment: (a) differences across persons in the attributes, response systems, and dimensions of a behavior problem, and (b) differences across persons in the causal relations relevant to a particular behavior problem. Because of these individual differences, nomothetic measures often reflect variance that is irrelevant to the targeted construct for the individual. We present a psychometric framework for idiographic assessment by first summarizing why the psychometric principles used to develop standardized measures of nomothetic constructs can create incongruity between the nomothetic measure and the characteristics of the targeted construct for an individual. We then develop a psychometric framework for idiographic assessment that combines components of multilevel modeling (random effects) and confirmatory factor analyses applied to repeated measurements of each individual. We also provide a step-by-step guide for the development and evaluation of an idiographic assessment instrument. PMID- 19217702 TI - Association of body mass index with decline in residual kidney function after initiation of dialysis. AB - BACKGROUND: Obesity is a risk factor for loss of kidney function in the general population, but it is unknown whether it proceeds to affect residual kidney function when patients require dialysis. Our aim was to study the effects of body mass index (BMI) on decline in kidney function and risk to develop anuria after initiation of dialysis therapy. STUDY DESIGN: Prospective cohort study. SETTING & PARTICIPANTS: 1,271 incident dialysis patients from 38 centers in The Netherlands participating in the Netherlands Cooperative Study on the Adequacy of Dialysis (NECOSAD) between 1997 and 2006. PREDICTOR: BMI assessed at 3 months after the initiation of dialysis therapy (baseline) and categorized into 4 groups: less than 20, 20 or greater to 25, 25 or greater to 30, and 30 or greater kg/m(2). OUTCOMES & MEASUREMENTS: The decrease in measured glomerular filtration rate (mGFR) was determined by means of linear mixed models and adjusted for age, sex, primary kidney disease, dialysis modality, smoking, cardiovascular disease, and normalized protein nitrogen appearance and additionally for proteinuria, blood pressure, and baseline mGFR. Cox regression analysis was used to calculate hazard ratios for the development of anuria. RESULTS: Patients had a mean age of 59 +/- 15 years, BMI of 24.8 +/- 4.1 kg/m(2), and mGFR of 4.7 +/- 3.3 mL/min. During 18 months of follow-up, the decrease in mGFR in patients with normal weight was 1.2 mL/min/y (95% confidence interval [CI], 0.7 to 1.6). Compared with those values, adjusted losses of mGFR were 0.4 mL/min/y (95% CI, 0.02 to 0.8) greater for overweight and 1.2 mL/min/y (95% CI, 0.5 to 1.8) greater for obese patients. In contrast, the decrease in underweight patients was 0.6 mL/min/y (-0.1 to 1.3) less. Anuria occurred in 297 patients; the risk was similar among BMI groups after adjustment for confounders and baseline diuresis. LIMITATIONS: Patients with missing BMI or mGFR values at baseline were excluded. CONCLUSION: Obesity is a strong risk factor for the decline in kidney function after initiation of dialysis therapy. Whether obese dialysis patients might benefit from a healthy weight reduction needs to be studied further. PMID- 19217704 TI - Pathways connecting neighborhood influences and mental well-being: socioeconomic position and gender differences. AB - Few studies have explored how participant socioeconomic position (SEP) and gender is related to perceptions of the pathways connecting neighborhood influences and mental well-being. This research used the concept mapping method, an intensive structured conceptualization process that produces pictorial views of how concepts are connected and interrelated. Thirty-six low and non-low SEP men and women from Toronto, Canada participated in the concept mapping sessions. One hundred and twenty unique neighborhood characteristics were felt to be related to mental well-being and those items were grouped into six distinct clusters. Notable differences in cluster importance by participant SEP status were found. While no overall differences were observed for males verses females, further stratification by both participant SEP and gender suggested that gendered perceptions are not uniform. Participant-created diagrams illustrated how the cluster domains are related to each other and to good mental well-being. These findings are important for uncovering the mechanisms by which neighborhoods differentially affect the mental health of residents from different SEPs and genders. PMID- 19217705 TI - [Sensibility of the abdomen after high superior tension abdominoplasty]. AB - INTRODUCTION: Patients who undergo an abdominoplasty frequently complain about the loss of sensibility of the abdominal wall. In this study, we analyze this sensibility after the high tension abdominoplasty. PATIENTS AND METHODS: This is a prospective study of 23 females operated between July 2003 and January 2005. The abdominoplasty technique used in our study combines extensive liposuccion, limited undermining centered on the linea alba and traction sutures. The sensibilty tests are carried out preoperatively, as well as at 3 and 6 months postoperatively. The three components of the skin sensibility -tactile, algesic and thermic- are evaluated in four differents areas of the abdomen. RESULTS: In the lateral areas of the abdomen (liposucted only), the thermoalgesic sensibility is diminished at 3 months and completely recovers at 6 months. At 3 months postoperatively, the tactile sensibility is even better than the preoperative one and continues to improve by 6 months. The postoperative hypogastric area is widely undermined during surgery. In this area, the three types of sensibility are heavily altered at 3 months and only partially recover at 6 months. The undermining of the postoperative epigastric area is limited. In this zone, the postoperative thermoalgesic sensibility is diminished at 3 months, but completely recovers at 6 months. At 3 months, the tactile sensibility is less than the preoperative one, but it improves with time to even exceed the preoperative values at 6 months. CONCLUSION: The high tension abdominoplasty only needs a limited undermining and largely preserves the innervation of the abdominal flap. Only the hypogastric area, largely undermined, presents a sensitivity loss. These results are better than those previously reported in the literature. PMID- 19217706 TI - Cerebral effects of three resuscitation protocols in uncontrolled haemorrhagic shock: a randomised controlled experimental study. AB - BACKGROUND: To compare haemodynamic and cerebral variables during aggressive fluid resuscitation vs. administration of a hypertonic starch solution (HS) combined with either noradrenaline [norepinephrine] or arginine vasopressin in an animal model of uncontrolled haemorrhagic shock. METHODS: After Animal Investigational Committee approval, 24 anaesthetised pigs underwent a liver trauma. At haemodynamic decompensation, animals were randomly assigned to receive fluid resuscitation (6% HES 130/0.4, 20 mL/kg, and Ringer, 40 mL/kg; FR group, n=8), or noradrenaline (bolus 20 microg/kg, continuously 1 microg/kg/min) combined with HS (7.2% NaCl/6% HES 200/0.5; 4 mL/kg) (n=8; NA/HS group), or vasopressin (bolus 0.2U/kg, continuously 0.04 U/kg/min) combined with HS (4 mL/kg) (n=8; AVP/HS group), respectively. Thirty minutes after drug administration, bleeding was controlled manually. RESULTS: Mean arterial blood pressure (MAP), cerebral perfusion pressure (CPP), and brain tissue oxygen pressure (P(bt)O(2)) decreased significantly with haemorrhage in all groups (p<0.05). AVP/HS resulted in a faster and higher increase of MAP and CPP compared to both NA/HS and FR (p<0.001 vs. FR; p<0.01 vs. NA/HS). Compared to FR, P(bt)O(2) increased faster with AVP/HS and NA/HS (p<0.05) after therapy, and ICP was lower at the end of the study period (p<0.05). All animals (8/8) of the AVP/HS group survived, compared to 4/8 and 4/8 in the NA/HS and FR group, respectively (p=0.07). CONCLUSIONS: Following uncontrolled haemorrhagic shock in this animal model, combination of HS with arginine vasopressin increased CPP and cerebral oxygenation faster than aggressive fluid resuscitation, without re increasing ICP. PMID- 19217707 TI - Amniotic fluid insulin-like growth factor binding protein 3 concentration as early indicator of fetal growth restriction. AB - OBJECTIVE: Insulin-like growth factor-I (IGF-I) is an important regulator of fetal growth and its bioavailability depends on insulin-like growth factor binding proteins (IGFBPs). Genes coding for IGF-I and IGFBP3 are polymorphic. We hypothesized that either amniotic fluid protein concentration at the beginning of the second trimester or genotype of one of these two genes could be predictive of abnormal fetal growth. STUDY DESIGN: Amniotic fluid samples (14-18 weeks of pregnancy) from 123 patients with appropriate for gestational age (AGA) fetuses, 39 patients with small for gestational age (SGA) fetuses and 34 patients with large for gestational age (LGA) were analyzed. Protein concentrations were evaluated by ELISA and gene polymorphisms by PCR. RESULTS: Amniotic fluid IGFBP3 concentrations were significantly higher in SGA compared to AGA group (P=0.030), and this was even more significant when adjusted to gestational age at the time of amniocentesis and other covariates (ANCOVA analysis: P=0.009). Genotypic distribution of IGF-I variable number of tandem repeats (VNTR) polymorphism was significantly different in SGA compared to AGA group (P=0.029). 19CA/20CA genotype frequency was threefold decreased in SGA compared to AGA group and the risk of SGA occurrence of this genotype was decreased accordingly: OR=0.289, 95%CI=0.1-0.9, P=0.032. Genotype distribution of IGFBP3(A-202C) polymorphism was similar in all three groups. CONCLUSIONS: High IGFBP3 concentrations in amniotic fluid at the beginning of the second trimester are associated with increased risks of SGA while 19CA/20CA genotype at IGF-I VNTR polymorphism is associated with reduced risks of SGA. Neither IGFBP3 concentrations, nor IGF-I/IGFBP3 polymorphisms are associated with modified risks of LGA. PMID- 19217708 TI - Identification of a novel estrogen receptor beta1 binding partner, inhibitor of differentiation-1, and role of ERbeta1 in human breast cancer cells. AB - Estrogen plays an important role in the proliferation and progression of breast cancer. The estrogen signal is mediated by the estrogen receptors (ERalpha and ERbeta). ERalpha (estrogen receptor alpha) is an important promoter of growth in breast cancer; however, the role of ERbeta (estrogen receptor beta) in breast cancer is less clear. In this study, using a yeast two-hybrid screening technique, we identified a novel ERbeta1-interacting protein, inhibitor of differentiation-1 (Id1), which is a dominant negative regulator of bHLH transcription factors, and promotes cell proliferation in breast cancer cells. Using mammalian two-hybrid protein-protein interaction assays, we found that the helix-loop-helix domain of the Id1 protein was essential for the physical interaction between ERbeta1 and Id1. In addition, we found that 17-beta estradiol inhibits ERbeta1 binding with Id1. Furthermore, we observed that ERbeta1 inhibited cell growth of MDA-MB-231 cells and upregulated p21 expression and that ERbeta1 up-regulation of p21 is Id1 dependent. Taken together, our study demonstrates a novel ERbeta1 binding partner, Id1, and a mechanism by which ERbeta1 inhibits breast cancer cell growth through binding with Id1 and upregulating p21 gene expression. PMID- 19217709 TI - Role of p53 in the induction of cyclooxygenase-2 by cisplatin or paclitaxel in non-small cell lung cancer cell lines. AB - Non-small cell lung Cancer (NSCLC) is extremely resistant to chemotherapeutic agents, such as cisplatin. High expression of the inflammatory enzyme cyclooxygenase-2 (COX-2) has been shown to inhibit chemotherapy-induced apoptosis, but little is known about COX-2 regulation upon drug treatment. Recent data indicate the tumor suppressor protein p53 as an important regulator of COX 2. Therefore, TP53 status could change tumor sensitivity to chemotherapy through induction of the anti-apoptotic protein COX-2. The main objective of this work was to analyze the effect of chemotherapy on the expression of COX-2, according to TP53 status. We report herein that lung cancer cell lines expressing wild-type p53, when exposed to cisplatin treatment, induced COX-2 (mRNA and protein), with concurrent synthesis of prostaglandins (PGE(2)). In contrast, COX-2 expression was not changed after cisplatin treatment of cells containing an inactive form of p53. Further, after silencing of wild-type p53 expressed in A549 cells by RNA interference, cisplatin was no longer able to induce COX-2 expression. Therefore, we suggest that induction of COX-2 by cisplatin in NSCLC cell lines is dependent on p53. For paclitaxel treatment, an increase in COX-2 mRNA expression was observed in H460 and A549 (wild-type p53 cell lines). Moreover, paclitaxel treatment increased COX-2 expression in ACC-LC-319 cell lines (p53 null), showing a p53-independent effect. These data may have therapeutic implications in the selection of patients and strategy for future COX-2 inhibition trials. PMID- 19217710 TI - DNA damage response of A549 cells treated with particulate matter (PM10) of urban air pollutants. AB - We describe the events triggered by a sub-lethal concentration of airborne particulate matter (PM(10)) in A549 cells, which include the formation DNA double strand breaks, gammaH2A.X generation, and 53BP1 recruitment. To protect the genome, cells activated ATM/ATR/Chk1/Chk2/p53 pathway but, after 48 h, cells turned into a senescence-like state. Trolox, an antioxidant, was able to prevent most of the alterations observed after particulate matter exposure, demonstrating the important role of ROS as mediator of PM(10)-induced genotoxicity and suggesting that DNA damage could be the mechanisms by which particulate matter augment the risk of lung cancer. PMID- 19217711 TI - Molecular based equation of state for shocked liquid nitromethane. AB - An approach is proposed to obtain the equation of state of unreactive shocked liquid nitromethane. Unlike previous major works, this equation of state is not based on extended integration schemes [P.C. Lysne, D.R. Hardesty, Fundamental equation of state of liquid nitromethane to 100 kbar, J. Chem. Phys. 59 (1973) 6512]. It does not follow the way proposed by Winey et al. [J.M. Winey, G.E. Duvall, M.D. Knudson, Y.M. Gupta, Equation of state and temperature measurements for shocked nitromethane, J. Chem. Phys. 113 (2000) 7492] where the specific heat C(v), the isothermal bulk modulus B(T) and the coefficient of thermal pressure (deltaP/deltaT)(v) are modeled as functions of temperature and volume using experimental data. In this work, we compute the complete equation of state by microscopic calculations. Indeed, by means of Monte Carlo molecular simulations, we have proposed a new force field for nitromethane that lead to a good description of shock properties [N. Desbiens, E. Bourasseau, J.-B. Maillet, Potential optimization for the calculation of shocked liquid nitromethane properties, Mol. Sim. 33 (2007) 1061; A. Hervouet, N. Desbiens, E. Bourasseau, J. B. Maillet, Microscopic approaches to liquid nitromethane detonation properties, J. Phys. Chem. B 112 (2008) 5070]. Particularly, it has been shown that shock temperatures and second shock temperatures are accurately reproduced which is significative of the quality of the potential. Here, thermodynamic derivative properties are computed: specific heats, Gruneisen parameter, sound velocity among others, along the Hugoniot curve. This work constitutes to our knowledge the first determination of the equation of state of an unreactive shocked explosive by molecular simulations. PMID- 19217712 TI - Biosurfactant-enhanced removal of total petroleum hydrocarbons from contaminated soil. AB - A screening method was developed to evaluate the oil removal capability of biosurfactants for oil-contaminated soils collected from a heavy oil-polluted site. The ability of removing total petroleum hydrocarbon (TPH) from soil by two biosurfactants was identified and compared with that of synthetic surfactants. The results show that biosurfactants exhibited much higher TPH removal efficiency than the synthetic ones examined. By using 0.2 mass% of rhamnolipids, surfactin, Tween 80, and Triton X-100, the TPH removal for the soil contaminated with ca. 3,000 mg TPH/kg dry soil was 23%, 14%, 6%, and 4%, respectively, while removal efficiency increased to 63%, 62%, 40%, and 35%, respectively, for the soil contaminated with ca. 9000 mg TPH/kg dry soil. The TPH removal efficiency also increased with an increase in biosurfactant concentration (from 0 to 0.2 mass%) but it did not vary significantly for the contact time of 1 and 7 days. PMID- 19217714 TI - Applicability of the Sludge Biotic Index (SBI) for MBR activated sludge control. AB - In 1994, Madoni created the Sludge Biotic Index (SBI) based on the presence and abundance of certain key protozoan groups. The use of this index has become increasingly common in wastewater treatment plant control and its applicability has been demonstrated. In this study, we applied the SBI index to a pilot-scale membrane bioreactor equipped with polyvinylidenefluoride hollow fibre membranes (0.04 microm pore size). Two different sludge retention times (SRT)--25 and 35 days--were assayed, with a constant hydraulic retention time of 30h. Operational conditions for MBR cause the fragmentation of flocs which are broken down into small dissipated elements with abundant dispersed bacteria. This process determines protozoan composition and colonization of the activated sludge. Results of the experiment showed a constant predominance of small flagellates, carnivorous ciliated protozoa and rotifers, above all in assays with 35-day SRT, independently of effluent quality. However, continuous changes in microbiota were observed, with a clear tendency for the SBI to increase over time as the sludge became more stable. Therefore, although it was not possible to establish an association between SBI and effluent quality for MBR activated sludge, the stabilisation of the process may be related to SBI. PMID- 19217713 TI - Thermocatalytic destruction of gas-phase perchloroethylene using propane as a hydrogen source. AB - The use of propane in combination with oxygen to promote the destruction of perchloroethylene (PCE) over a platinum (Pt)/rhodium (Rh) catalyst on a cerium/zirconium oxide washcoat supported on an alumina monolith was explored. Conversions of PCE were measured in a continuous flow reactor with residence times less than 0.5s and temperatures ranging from 200 to 600 degrees C. The presence of propane was shown to increase significantly the conversion of PCE over oxygen-only conditions. Conversions close to 100% were observed at temperatures lower than 450 degrees C with 20% oxygen and 2% propane in the feed, which makes this process attractive from a practical standpoint. In the absence of oxygen, PCE conversion is even higher, but the catalyst suffers significant deactivation in less than an hour. Even though results show that oxygen competes with reactants for active sites on the catalyst, the long-term stability that oxygen confers to the catalyst makes the process an efficient alternative to PCE oxidation. A Langmuir-Hinshelwood competitive adsorption model is proposed to quantify PCE conversion. PMID- 19217715 TI - Copper fractionation and release from soils devoted to different crops. AB - We studied copper release as a function of pH in eight samples of acid soils under vine and maize crops containing variable concentrations of total copper (Cu(t)) from 55 to 112 mg kg(-1). Although both types of soil had similar Cu(t) values, the origin of the copper was essentially anthropogenic in the vineyard soils and natural (as result of weathering of parent material) in those under maize. The disparate origin of the metal resulted in marked differences in its fractions, particularly as regards organically bound copper, which was much more abundant in the vineyard soils. This had a strong effect on copper release as a function of pH, which was one order of magnitude greater in the vineyard soils than in those under maize. PMID- 19217716 TI - Lead in edible mushrooms: levels and bioaccumulation factors. AB - Lead content was determined in 238 samples of 28 species of edible mushrooms collected from different sites in the province of Lugo (NW Spain) during 2005 and 2006. The hymenophore (H) and the rest of the fruiting body (RFB) were analysed separately. The analyses were carried out by an anodic stripping voltammetric technique using drop mercury as the working electrode. The highest mean lead contents (mg/kg dry weight) of 3.6 and 4.1, 3.0 and 2.2, 2.5 and 2.3, 2.4 and 2.3 were determined in Coprinus comatus, Agaricus campestris, Lepista nuda and Calvatia utriformis in hymenophore and the rest of fruiting bodies, respectively, while the lowest in Agaricus bisporus (0.35 in H and 0.54 in RFB) and Fistulina hepatica (0.41 in H and 0.50 in RFB). All mushroom species were bioexclusors of lead (BCF<1) in relation to the underlying soils. There were not statistically significant differences between lead levels in hymenophore and in the rest of the fruiting bodies. The lead concentrations were compared to literature data and levels set by legislation. It can be concluded that the consumption of these mushrooms can not be considered as a toxicological risk from lead content point of view, and they provide a nutritional requirement to the diet. PMID- 19217717 TI - Cost effectiveness of two rehabilitation programmes for neck and back pain patients: A seven year follow-up. AB - The cost effectiveness of work-oriented rehabilitation for persons on long-term sick leave needs to be assessed. This prospective observational study presents a follow-up seven years after rehabilitation using two different evidence-based work-oriented regimens. Individuals on sick leave for neck and back pain were referred to two rehabilitation programmes in Sweden. The first programme was a relatively low-intensity programme based on orthopaedic manual therapy and exercise programme (OMTP). The second programme was a full-time multidisciplinary programme (MDP). The primary outcome was sickness absence seven years after intervention. Cost effectiveness was calculated on the basis of loss of production due to sickness absence. The results show that participants referred to MDP and with less than 60 sick days before rehabilitation have reduced sickness absence after intervention as compared to matched controls. This corresponds to a cost reduction of about 94,494 EUR per referred individual. Further, the results indicate that participants of the OMTP who have more than 60 sick days before rehabilitation have a statistically significant increased risk of disability pension. This means increased cost in terms of loss of production of 44,593 EUR per referred individual. The results of this study show that MPD but not OMTP achieves the goal of working life-oriented rehabilitation. A direct comparison between the rehabilitation programmes strengthened the assumption that long-term sickness absence prior to rehabilitation is associated with more days on sick leave after rehabilitation. This analysis also indicated the importance of participants' pain self-efficacy beliefs and recovery beliefs on rehabilitation outcome. PMID- 19217718 TI - Image simulation of high resolution energy filtered TEM images. AB - Inelastic image simulation software is presented, implementing the double channeling approximation which takes into account the combination of multiple elastic and single inelastic scattering in a crystal. The approach is described with a density matrix formalism. Two applications in high resolution energy filtered (EFTEM) transmission electron microscopy (TEM) images are presented: thickness-defocus maps for SrTiO(3) and exit plane intensities for an (LaAlO(3))(3)(SrTiO(3))(3) multilayer system. Both systems show a severe breakdown in direct interpretability which becomes worse for higher acceleration voltages, thicker samples and lower excitation edge energies. Since this effect already occurs in the exit plane intensity, it is a fundamental limit and image simulations in EFTEM are indispensable just as they are indispensable for elastic high resolution TEM images. PMID- 19217719 TI - Co-infection of chickens with Eimeria praecox and Eimeria maxima does not prevent development of immunity to Eimeria maxima. AB - Previous studies revealed an ameliorating effect of Eimeria praecox on concurrent E. maxima infection, such that weight gain, feed conversion ratio, and intestinal lesions were nearly identical to uninfected or E. praecox-infected controls. The purpose of the present study was to determine if protective immunity against E. maxima challenge infection developed in chickens infected with both E. praecox and E. maxima. Day-old chickens were infected with 10(3)E. praecox, 10(3)E. maxima, or a mixture of 10(3)E. praecox and 10(3)E. maxima oocysts. Chickens were then challenged at 4 weeks of age with 5x10(4)E. praecox or 5x10(3)E. maxima oocysts and clinical signs of coccidiosis were assessed 7 days post-challenge. Relative to non-challenged controls, naive chickens or chickens immunized with E. praecox displayed a 32-34% weight gain depression after challenge with 5x10(3)E. maxima oocysts. In contrast, chickens immunized with either E. maxima oocysts alone or a combination of E. praecox and E. maxima oocysts displayed complete protection against lower weight gain associated with E. maxima challenge. Also, protection against decreased feed conversion ratio and intestinal lesions was observed in single E. maxima- or dual E. maxima+E. praecox-immunized chickens. These findings indicate that co-infection of chickens with E. maxima and E. praecox does not prevent development of immunity against E. maxima or E. praecox challenge. PMID- 19217720 TI - Associations between smoking cessation and anxiety and depression among U.S. adults. AB - Many studies have shown a relationship between smoking and depression. However, few studies have examined the association between current depression and smoking and even fewer used large cross-sectional data to support these findings. Using the 2006 Behavioral Risk Factor Surveillance System data (n=248,800), we compared rates of lifetime depression, lifetime anxiety, current depression, and current depressive symptoms among smokers who unsuccessfully attempted to quit (unsuccessful quitters), former smokers (successful quitters), and smokers who made no attempts to quit (non-quitters). Unsuccessful quitters experienced more lifetime depression and anxiety than non-quitters (OR=1.2; 95% CI, 1.0-1.4), whereas successful quitters experienced less (OR=0.7, 95% CI, 0.6-0.8). Current depression prevalence was 14.3% among non-quitters, 18.8% among unsuccessful quitters, and 8.0% among successful quitters. On average, unsuccessful quitters also experienced more days of depressive symptoms during the previous month than either non-quitters or successful quitters. Our results suggest that smokers who attempt to quit unsuccessfully may experience lifetime depression as well as current depression at a higher rate than other smokers and former smokers. PMID- 19217721 TI - Biopsies of colorectal clinical polyps--emergence of diagnostic information on deeper levels. AB - Although the occasional appearance of a normal histology of biopsies from endoscopic colorectal (CR) polyps is generally held knowledge, its prevalence has rarely been focused on, and the yield of additional sections in such cases has been previously addressed in merely four communications. Hitherto, this issue has not been discussed in the context of the clinical setting. The prime aim of this study was to evaluate the yield of step sectioning CR biopsies, considered non diagnostic (non-diagnostic biopsies (NDB)) on routine sections. The results are correlated with the indications for endoscopy. Additionally, an appropriate, cost effective approach for handling NDB was sought. Biopsies from 480 clinical polyps were prospectively evaluated by one of three gastrointestinal pathologists and classified as (a) diagnostic biopsies (DB), comprising neoplastic polyps, hyperplastic polyps (HP), sessile-serrated polyp, other diverse causes of polyp formation and (b) NDB comprising normal histology (group 1), suspicious of either adenoma (group 2) or HP (group 3). Material grouped 1-3 was subsequently step sectioned (three sections prepared from each of nine additional levels). The biopsy specimens were obtained from 245 endoscopies and stratified in the following categories according to the clinical indications: relevant symptoms (symptomatic, n=127), previously documented sporadic large bowel neoplasia (follow-up, n=99), and documented or presumed hereditary condition that confer an increased risk of CRC (hereditary, n=19, including 15 hereditary non-polyposis colorectal cancer (HNPCC) cases). Sixty-five (13.5%) of the 480 samples were classified as NDB (normal morphology n=49, suspicious of adenoma n=5, suspicious of HP n=11), constituting roughly 10% of all biopsies from the symptomatic and the follow-up categories, 32.1% of samples from the hereditary cases, the difference between the hereditary and the non-hereditary cases being statistically significant (p<0.0001). Upon leveling the 65 NDB, a DB emerged in 24 (36.9%) cases, with no significant difference in the yield in relation to the delineated indication categories. Thereby, diagnostic information was obtained with three additional levels in 15 cases, the remaining 9 cases requiring additional sections, ranging from 4 to 8 levels. The present step sectioning approach implied an extra expense of about 112 US$ for each NDB converted to a DB. The higher prevalence of NDB in relation to genetic disorders probably reflects sampling of particularly diminutive lesions. Given the high yield of step sectioning NDB coupled with an acceptable price, our strategy delineated here is recommended in routine practice with the modification of an initial preparation of sections from merely three levels, and if still non-diagnostic, supplementation with additional five levels. PMID- 19217722 TI - Gender effect on association between DRD2 polymorphism and substance dependence in a Spanish sample. AB - Our aim was to examine a possible association between substance dependence and the TaqIA polymorphism of the D2 dopamine receptor (DRD2), a single nucleotide polymorphism (SNP) located at the 3' UTR region of the DRD2 gene. A case-control design stratified by gender was used to analyze the genotypes of this SNP in a sample of 125 substance-dependent patients according to DSM-IV and 203 blood donors recruited as controls in two general city hospitals in Madrid, Spain. Genomic DNA from peripheral blood samples was amplified through PCR to identify the variants of the SNP in the DRD2 gene. Analyses performed with Chi(2) tests revealed that the A1 allele (A1/A1 and A1/A2 genotypes) of the Taq 1A SNP of the DRD2 gene was significantly associated with substance dependence in males, but not in the whole sample. Male patients had significantly higher rates of the A1 containing genotypes than male controls. The finding of an association between substance dependence and the DRD2 gene TaqIA SNP only in males suggests the existence of gender-specific differences in the genetic underpinnings of substance dependence. PMID- 19217723 TI - Effects of cocaine esterase following its repeated administration with cocaine in mice. AB - BACKGROUND: A bacterial cocaine esterase (CocE) produces robust protection and reversal of cocaine toxicity. The aim of this study was to investigate how effectiveness of CocE was changed following its repeated administration together with cocaine. METHODS: Cocaine toxicity was quantified by measuring the occurrence of convulsions and lethality in mice. Immunologic responses of CocE were determined using ELISA. In the protection experiment, i.v. CocE 0.3mg was given 1min before a lethal dose of i.p. cocaine 180mg/kg. In the rescue experiment, i.v. CocE 0.3mg was given 1min after the occurrence of convulsions elicited by i.p. cocaine 100mg/kg. In both treatment paradigms, four trials were conducted in the same animals with a 2-week interval. RESULTS: CocE retained its effectiveness to protect or rescue mice during the first two trials and these mice did not show an immune response. In contrast, CocE's effectiveness was gradually reduced in the last two trials, accompanied by 10- and 100-fold increases in anti-CocE antibody titers. Nevertheless, effectiveness of CocE could be partially recovered by increasing the dose of CocE. In addition, escalating the dose of CocE from the minimum effective dose for repeated administration could also retain CocE's effectiveness longer and slow the production of anti CocE antibodies. CONCLUSIONS: These results indicate that CocE is a weak antigen and it can maintain its protective and rescuing ability initially against cocaine induced toxicity. Decreased effectiveness of CocE following repeated use can be partially improved by adjusting the dose and frequency of CocE treatment. PMID- 19217724 TI - Reliability and validity of a short form of the Marijuana Craving Questionnaire. AB - BACKGROUND: The Marijuana Craving Questionnaire (MCQ) is a valid and reliable, 47 item self-report instrument that assesses marijuana craving along four dimensions: compulsivity, emotionality, expectancy, and purposefulness. For use in research and clinical settings, we constructed a 12-item version of the MCQ by selecting three items from each of the four factors that exhibited the greatest within-factor internal consistency (Cronbach's alpha coefficient). METHODS: Adult marijuana users (n=490), who had made at least one serious attempt to quit marijuana use but were not seeking treatment, completed the MCQ-Short Form (MCQ SF) in a single session. RESULTS: Confirmatory factor analysis of the MCQ-SF indicated good fit with the 4-factor MCQ model, and the coefficient of congruence indicated moderate similarity in factor patterns and loadings between the MCQ and MCQ-SF. Homogeneity (unidimensionality and internal consistency) of MCQ-SF factors was also consistent with reliability values obtained in the initial validation of the MCQ. CONCLUSIONS: Findings of psychometric fidelity indicate that the MCQ-SF is a reliable and valid measure of the same multidimensional aspects of marijuana craving as the MCQ in marijuana users not seeking treatment. PMID- 19217725 TI - Distribution of superantigenic toxin genes in Staphylococcus aureus isolates from milk samples of bovine subclinical mastitis cases in two major diary production regions of China. AB - To evaluate the distribution of most known staphylococcal superantigen (SAg) genes in Staphylococcus aureus isolated from bovine mastitis cases, a genetic analysis of 15 SAg genes and genotypes was performed in a total of 283 S. aureus isolates collected from milk samples of cows with subclinical mastitis in two major diary production regions of China. Almost 65% of the isolates possessed at least one toxin gene. The most frequently found genes were sea (36.0%) followed by sei (31.8%), seg (31.4%) and selm (26.9%). The genes see, selk, or selo were not found in any of the isolates tested. Overall, 28 SAg genotypes were observed, among which the genotypes sea-seg-sei-selm, seg-sei-selm-seln, and sea-sed-selj predominated at the rate of 8.8%, 7.4%, and 6.7%, respectively. Marked geographical variations were noticed in the distribution of individual SAg genes and genotypes among S. aureus isolates from the two different regions. The relationship between toxin genotypes and toxin genes encoding profiles of mobile genetic elements (MGEs) was analyzed, revealing that majority of SAg genes were present in certain MGEs, which were in accordance with current knowledge about MGEs carrying those genes. However, some gene combinations suggest the possibility of the existence of variants or new types of MGEs. PMID- 19217726 TI - Therapeutic management of patent ductus arteriosus. AB - Management of patent ductus arteriosus (PDA) in premature neonates has always been a challenge to the neonatologist and continues to be a controversial topic. Indications for treatment are not clear. Approximately 40% of ductus arteriosus close spontaneously even in extremely-low-birth-weight neonates. Prophylactic or early pre-symptomatic treatment may unnecessarily expose these babies, in whom the ductus might close spontaneously, to pharmacologic agents and their adverse effects. On the other hand, with advancing postnatal age, delaying treatment could potentially decrease successful medical closure thereby increasing the rate of surgical ligation and the complications associated with surgery. We tried to develop some clinical guidelines for management of PDA in premature neonates based on evidence, or lack thereof, and experience. PMID- 19217727 TI - Immaturity, perinatal inflammation, and retinopathy of prematurity: a multi-hit hypothesis. AB - OBJECTIVE: To explore the relationship among markers of infection/inflammation in their association with retinopathy of prematurity (ROP). METHODS: We studied clinical characteristics and 4 single nucleotide polymorphisms in infection/inflammation-associated genes in a group of 73 children with a gestational age<32 weeks. Forty-four children (60%) had ROP, of whom 13 (30% of those with ROP) progressed to stage 3 ROP. No child had grade 4 or 5 ROP. We employed both descriptive and analytic statistical methods. RESULTS: Clinical variables of infection/inflammation were consistently associated with an increased risk of ROP. Among infants with ROP, they were also associated with progression to ROP grade 3. Genetic markers were not associated with ROP occurrence, but with progression to high grade disease. In tri-variable analyses exploring the effects of gestational age <29 weeks, clinical chorioamnionitis (CAM) and neonatal systemic inflammatory response syndrome (SIRS) on ROP occurrence, low gestational age was the most important antecedent, while additional individual or joint exposure to SIRS and CAM add appreciably to this risk of progression to high grade disease. CONCLUSION: Both antenatal and neonatal exposure to inflammation appear to contribute to the increased ROP risk in preterm infants. PMID- 19217728 TI - Is the Menopause Rating Scale accurate for diagnosing sexual dysfunction among climacteric women? AB - BACKGROUND: Although several tools have been designed to assess quality of life (QoL) among middle-aged women their capacity to specifically assess sexual dysfunction (SD) remains uncertain. Moreover, if SD impairs QoL within this population, then sexual assessment becomes a key issue. OBJECTIVES: To evaluate the accuracy of the Menopause Rating Scale (MRS) in diagnosing SD among climacteric women. METHODS: In this cross-sectional study 370 women aged 40-59 years filled out the MRS and the Female Sexual Functioning Index (FSFI) simultaneously. SD among surveyed women was defined as those obtaining a total FSFI score of or=1 in the MRS item 8 as a cut-off value for discriminating women with SD (78% sensitivity and 62% specificity with an area below the curve of 0.70 Swett). CONCLUSIONS: The MRS was moderately accurate for diagnosing SD among climacteric women. More research is warranted in this regard. PMID- 19217729 TI - A secret pathway of the sinus node artery. AB - Anatomic and postmortem angiographic findings of a previously unreported case of common origin of the left circumflex (LCX) and the sinus node (SN) arteries, from the left main (LM) coronary artery were demonstrated. Knowledge of this anatomical variation, although it does not give rise to symptoms, is essential for anatomist and mainly for the interventional cardiologists and cardiac surgeons for their procedures. PMID- 19217730 TI - Fatal cytisine intoxication and analysis of biological samples with LC-MS/MS. AB - We report about a fatal cytisine intoxication in a 20-year-old man who, according to his mother, had drunken tea prepared from plant material of Laburnum anagyroides with the toxic pyridine-like alkaloid as ingredient, which exhibits pharmacological effects similar to nicotine. Using a liquid chromatographic-mass spectrometric (LC-MS) procedure cytisine was quantified in post-mortem specimens. By exclusion of other causes of death an intoxication was determined as the cause of death with respiratory failure as the pathophysiological mechanism. PMID- 19217731 TI - Anaphylactic deaths in Maryland (United States) and Shanghai (China): a review of forensic autopsy cases from 2004 to 2006. AB - Anaphylaxis is a life threatening allergic reaction that is rapid in onset and may cause sudden death. A retrospective study was performed using the database from the Office of the Chief Medical Examiner (OCME) in the State of Maryland and the Department of Forensic Medicine in Shanghai Medical College (DFM-SMC) to examine the etiology and forensic investigation findings of anaphylactic deaths from 2004 to 2006. Details of the medical history, agent responsible for the allergic reaction, death scene investigation and postmortem examination findings were reviewed for all cases. A total of 28 cases of anaphylactic death were identified for the study period, 17 from Maryland and 11 from Shanghai. Of the 17 Maryland cases, 6 (35%) involved allergic reaction to food, 5 (25%) to drugs, 2 to bee stings, 1 to hair dye, and 3 to unknown allergens. Investigation revealed that 9 of 17 cases had a history of asthma and 8 had previous allergic reactions to certain foods and / or drugs. In Shanghai, all 11 deaths resulted from anaphylactic reaction to antibiotics, 10 of which occurred in clinics illegally operated by unlicensed physicians. The interval between the onset of symptoms and death ranged from less than 1 min to 3 days after initially contacting the allergen. In the majority (68%) of cases, death occurred within 8h after the onset of symptoms. Postmortem findings were relatively non-specific, and included pharyngeal/laryngeal edema 14/28 (50%), mucus plugging in the airways 11/28 (39%), and pulmonary congestion and edema 28/28 (100%). Serum tryptase concentrations were measured in 14 cases in Maryland and the concentration ranged from 3.3 ng/ml to 200 ng/ml. There were significant differences with regards to allergen type and the circumstances of death between these two regions. Inappropriate use of antibiotics and illegal medical practices were the main causes of identified anaphylactic death in Shanghai. In Maryland anaphylactic deaths were mainly caused by food reactions. This study indicates that the postmortem diagnosis of anaphylactic death is usually based on exclusion and circumstantial evidence. Knowledge of the patient's history and circumstances of death is of major importance for the forensic pathologist when investigating suspected anaphylactic death. PMID- 19217732 TI - Current trends in drug abuse associated fatalities - Jordan, 2000-2004. AB - This study is the first study that addresses drug abuse associated fatalities in Jordan. It is aimed to give a close picture to the demography, toxicological data, manner, cause of death and other associated findings in such cases. Postmortem forensic pathology reports for all autopsies examined in the National Institute of Forensic Medicine were reviewed over a 5-year period and drug abuse associated deaths were selected. The study revealed that 44 cases (0.76%) out of the 5789 total autopsies were attributed to drug abuse associated deaths. The age range was from 20 to 60 years (mean+/-S.D.=32.7+/-7.2). More than 80% of cases were Jordanian males. The reported abuse substances as single drug or in combination were alcohol in 56.8%, morphine 36.4%, heroin 15.9%, benzodiazepines in 11.4% and cocaine in one body packer case. Surprisingly, neither a case with amphetamine or amphetamine analogue, nor with marijuana or methadone was recorded. In 75% of cases the death was accidental and only one case was reported to be suicidal, while in 18.2% and 4.5% death was due to sudden death and road traffic accidents, respectively. Regarding the cause of death, it was related to drug overdose in 50% of cases and in 34.1%, 11.4% and 4.5% of cases it was attributed to drug related medical complications, non-drug related complications, and trauma, respectively. Alcohol was mainly associated with accidental death; morphine and heroin were associated with drug overdose and abused through intravenous route. Injection marks were reported in 56.8% of cases and in 52.3% death occurred at home. This study confirmed the variation in the incidence and type of abused substances in Jordan compared with different countries. PMID- 19217734 TI - Evaluation of non-isothermal methods in stability studies of human insulin pharmaceutical preparations. AB - The purpose of this research was to study the thermal stability of a human insulin pharmaceutical preparation using non-isothermal conditions and comparison with classical isothermal experiments. The isothermal studies were performed in the temperature range 20-60 degrees C, whereas non-isothermal stability studies were performed using a linear increasing temperature program, heating rate 0.25 degrees C per hour and temperature interval 30-70 degrees C. Under isothermal conditions, an apparent first-order degradation process was observed at all temperatures. The linear Arrhenius plot suggested that the insulin degradation mechanism was the same within the studied temperature range, with quite large uncertainties due to the small number of degrees of freedom based only on the scatter in the plot, giving an estimated shelf-life at 25 degrees C of 199.1 days. In non-isothermal conditions, the integral approach was used to estimate the activation parameters. It provides results in good agreement with those of the traditional method, but with the advantage that the uncertainty in the final result directly reflects the goodness of fit of the experimental data, since it takes into account the scatter in the original data. The estimated shelf-life in non-isothermal conditions was quite close to the value derived from isothermal data, 191.4 days, although the 95% confidence interval estimated were slightly higher. This is due to the differences in the estimation method and the nature of the experimental errors. The bootstrap technique is also applied to estimating confidence limits for the Arrhenius parameters and shelf-life. This method is very useful when the underlying distribution function of the parameters is unknown. The results obtained indicate that the Arrhenius parameters follow a normal distribution, whereas the shelf-life follows a log-normal distribution. In any case, the results obtained show that there is no difference between the asymptotic and bootstrap confidence intervals. PMID- 19217733 TI - The role of interleukin-1beta in febrile seizures. AB - Febrile seizures (FS) occur in children as a result of fever. Despite their prevalence, the pathophysiology of FS has remained unclear. Recent evidence from clinical and experimental studies has highlighted a potential role of immune generated products in the genesis of FS. Of particular interest are the pro inflammatory cytokine, interleukin-1beta (IL-1beta) and its naturally occurring antagonist, interleukin 1 receptor antagonist (IL-1ra). Using a novel animal model of FS, involving the generation of physiological fever, we investigated the role of the IL-1beta/IL-1ra system in the genesis of FS. We found that animals with FS had increased hippocampal and hypothalamic IL-1beta compared to equally treated animals without FS, which was first evident at onset of FS in the hippocampus. There were no differences in IL-1ra levels. ICV IL-1beta increased the number of animals with FS while IL-1ra had an opposite anti-convulsant effect. The data from these studies, in combination with recent results from other laboratories, have established a putative role for the IL-1beta/IL-1ra system in the genesis of FS. PMID- 19217735 TI - Development and characterization of a novel host cell DNA assay using ultra sensitive fluorescent nucleic acid stain "PicoGreen". AB - Development of a novel host cell DNA assay using PicoGreen is described, which is capable of detecting short double stranded DNAs (ds-DNAs) in cell culture supernatants and process intermediates. Examination of this PicoGreen DNA assay was carried out by determination of the DNA length detection limit, observation of short ds-DNAs in cell culture supernatants and process intermediates, evaluation of dose dependency and a supersensitizing protocol, and comparison of the novel assay with conventional assays for measuring host cell DNA concentration in real samples. The PicoGreen DNA assay was capable of detecting ds-DNAs as short as 20 bp, and the sensitivity of the PicoGreen DNA assay was comparable to that of the Threshold system with application of additional SDS/Proteinase K digesting and DNA concentrating steps. Also, the amount of DNA identified in both cell culture supernatant and process intermediates was clearly underestimated by the Threshold system results when compared with the PicoGreen DNA assay results. The PicoGreen DNA assay clearly provides better accuracy and is a simpler procedure for measuring host cell DNA levels in cell culture supernatant and process intermediates than the conventional method with the Threshold system. This newly developed DNA assay will be prominent among host cell DNA assays for measuring host cell DNA levels in bio-pharmaceuticals. PMID- 19217736 TI - Role of excipients in the quantification of water in lyophilised mixtures using NIR spectroscopy. AB - The ratio between mannitol and sucrose in a freeze-dried formulation has a major impact on the processing and the stability of a lyophilised product. The ratio of these common excipients influences a critical quality attribute of the system, namely the overall amount of water, due to the different nature of their water solid interactions. For this experiment samples containing various ratios of mannitol and sucrose and several other additives were freeze-dried, stored under different conditions and measured by NIR. Different spectral pre-processing methods and wavelength selections were tested. Multivariate analysis was applied to correlate the Karl Fischer titration to the NIR spectra. It was found that standard normal variate (SNV) transformation of the wavenumber range 4200-7400 cm(-1) yielded prediction errors close to the accepted measurement error of the Karl Fischer titration, when measuring samples of up to 5.5% (w/w) water. It was further found that there was a slight tendency for samples containing inorganic salts or histidine to be underestimated in the NIR measurements. However, no influence was found to be caused by the varying mannitol-sucrose ratio. By reducing the sample set to those samples containing up to around 2% of water, the error was found to be below the uncertainty originating from the reference method. Due to this it can no longer be determined whether the deviation originates from the NIR method or the reference method. It can therefore be concluded the NIR is a suitable tool for quantification of the water content in lyophilised samples with varying mannitol-sucrose ratios. PMID- 19217737 TI - Chromatographic separation and NMR characterization of the isomers of MMB-4, a bis-(pyridiniumaldoxime). AB - 1,1'-Methylenebis{4-[(hydroxyimino)methyl]pyridinium) dichloride (MMB-4), a promising antidote for organophosphate poisoning, has been shown by chromatography and NMR to be a mixture of geometric isomers, predominantly the E/E form. The chromatographically separated isomers have been isolated, directly characterized by NMR to be E/E and E/Z isomers of high purity, and shown by HPLC and NMR to re-equilibrate in solution to the isomeric mixture found in bulk MMB 4. These findings clearly show that a minor component in MMB-4 is not an impurity, but a geometric isomer of the principal component and demonstrate the need to understand equilibrium processes for drug characterizations and isomer distributions of chemicals proposed for animal and human clinical trials. Evidence for the presence of the Z/Z isomer could not be found. PMID- 19217738 TI - Simultaneous quantification of losartan and active metabolite in human plasma by liquid chromatography-tandem mass spectrometry using irbesartan as internal standard. AB - A simple and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method employing electronspray ionization was developed and validated for quantification of losartan and its carboxylic acid metabolite in human plasma using irbesartan as internal standard (IS). Following a simple pretreatment procedure, the analytes were separated using a gradient mobile phase on reverse phase C18 column. Selected reaction monitoring was specific for losartan, losartan acid and irbesartan. The method validation demonstrated the specificity, lower limit of quantification, accuracy and precision of measurements. The assay exhibited a linear dynamic range of 2.0-400 ng/mL for losartan and 1.85-370 ng/mL for losartan acid. A run time of 3.5 min for each sample made it possible to analyze more than 200 samples per day. The validated method has been successfully used to analyze human plasma samples for application in bioavailability/bioequivalence studies. PMID- 19217739 TI - Determination of alprostadil in rat plasma by ultra performance liquid chromatography-electrospray ionization-tandem mass spectrometry after intravenous administration. AB - A rapid, highly selective ultra performance liquid chromatography-electrospray ionisation-tandem mass spectrometry method (UPLC-ESI-MS/MS) was developed and validated for the determination and pharmacokinetic investigation of alprostadil in rat plasma. After a simple sample preparation procedure involving a one-step liquid-liquid extraction, alprostadil and the internal standard, diphenhydramine, were chromatographed on an ACQUITY UPLC BEH C(18) column with gradient elution using a mobile phase consisting of acetonitrile and water (containing 0.1% formic acid) at a flow rate of 0.25 mL min(-1). The detection was performed on a triple quadrupole tandem mass spectrometer in multiple reaction monitoring (MRM) mode via an electrospray ionization (ESI) source. The calibration curve was linear (r(2)=0.99) over the concentration range 0.4-250.0 ng mL(-1), with a lower limit of quantification of 0.4 ng mL(-1) for alprostadil. The inter- and intra-day precision (%R.S.D.) was less than 8.5% and 2.4%, respectively, and the accuracy (RE%) was between 9.3% and 1.0% (n=6). Alprostadil in rat plasma was stable when stored at room temperature for 0.5h and at -20 degrees C for two weeks. The method was very rapid, simple and reliable, and was employed for the first time for the pharmacokinetic studies of alprostadil in rats after a single intravenous administration of 50 microg kg(-1). PMID- 19217740 TI - Compatibility of atenolol with excipients: LC-MS/TOF characterization of degradation/interaction products, and mechanisms of their formation. AB - A study was carried out to investigate compatibility of atenolol, a beta(1) blocker, with a variety of pharmaceutical excipients. The binary mixtures (1:1) of atenolol with the excipients were stored for 1 month at 40 degrees C/75% RH. The samples were directly observed for the physical changes, and also analyzed by a validated HPLC method to determine the chemical changes. The study revealed that atenolol was incompatible with ascorbic acid, citric acid and butylated hydroxyanisole. The degradation/interaction products formed in these mixtures were characterized by high resolution mass spectrometric and fragmentation analyses, using a LC-MS/TOF system. The identity of characterized structures was justified through mechanistic explanations. PMID- 19217741 TI - Linking numeracy and asthma-related quality of life. AB - OBJECTIVE: To examine the correlation of numerical skills used in patients' self management of asthma with asthma-related quality of life (AQOL). METHODS: Adults with moderate-severe asthma completed the Asthma Numeracy Questionnaire (ANQ), assessments of reading comprehension and self-efficacy, and the mini-Asthma Quality of Life Questionnaire (miniAQLQ). The numeracy-AQOL relationship was evaluated in the context of potential confounders (demographic variables) and mediators (e.g. income and self-efficacy), using tests of correlation then multivariable models to assess for confounders and mediators. RESULTS: 80 adults with moderate or severe asthma were evaluated. Mean ANQ score was 2.3+/-1.2 (range 0-4). ANQ was correlated with miniAQLQ (rho=0.24, p=0.03). This association was sustained (rho=0.27, p=0.02) when controlling for potential confounders significantly associated with AQOL (age, Latino ethnicity). The ANQ miniAQLQ association was mediated by household income; the correlation was reduced by 81% when adjusting for income (rho=0.05, p=0.65). In contrast, self efficacy less strongly mediated this association; the correlation was reduced by 26% when controlled for self-efficacy (rho=0.20, p=0.08). CONCLUSION: Numerical skills needed for asthma self-management influence AQOL primarily through their impact on income and, to a lesser extent, on self-efficacy. PRACTICE IMPLICATIONS: Adults with asthma will benefit from self-management instructions employing the simplest mathematical constructs whose understanding is confirmed by clinicians. PMID- 19217742 TI - PowerON: the use of instant message counseling and the Internet to facilitate HIV/STD education and prevention. AB - OBJECTIVE: In recent years, Internet-based or online counseling has emerged as an effective way to assess psychological disorders and discuss destructive behaviors with individuals or groups of individuals. This study explores the application of online counseling to HIV/STD risk-taking behavior among men who have sex with men (MSM). METHODS: PowerON, an organization that provides sexual health information to MSM exclusively online, used instant message technology to counsel MSM in real time through computer-mediated means. A sample of 279 transcripts of instant message exchanges between PowerON counselors and Gay.com users were recorded and qualitatively analyzed. RESULTS: Approximately 43% of the instant message sessions discussed information about HIV/STD testing. Risk-taking behaviors were addressed in 39% of the sessions. Information about HIV/STDs and general counseling were given in 23% and 18% of the counseling sessions, respectively. CONCLUSION: The data showed these instant message sessions to be a potentially feasible forum for HIV/STD counseling. PRACTICE IMPLICATIONS: Information ordinarily disseminated at health clinics could be successfully distributed through the Internet to MSM. PMID- 19217743 TI - Vascular surgery training trends from 2001-2007: A substantial increase in total procedure volume is driven by escalating endovascular procedure volume and stable open procedure volume. AB - BACKGROUND: Endovascular procedure volume has increased rapidly, and endovascular procedures have become the initial treatment option for many vascular diseases. Consequently, training in endovascular procedures has become an essential component of vascular surgery training. We hypothesized that, due to this paradigm shift, open surgical case volume may have declined, thereby jeopardizing training and technical skill acquisition in open procedures. METHODS: Vascular surgery trainees are required to log both open and endovascular procedures with the Accreditation Council for Graduate Medical Education (ACGME). We analyzed the ACGME database (2001-2007), which records all cases (by Current Procedural Terminology [CPT] code) performed by graduating vascular trainees. Case volume was evaluated according to the mean number of cases performed per graduating trainee. RESULTS: The mean number of total major vascular procedures performed per trainee increased by 174% between 2001 and 2007 (from 298.3 to 519.2). Endovascular diagnostic and therapeutic procedures increased by 422% (from 63.7 to 269.1) and accounted for 93.0% of the increase in total procedures. The number of open aortic procedures (aneurysm, occlusive, mesenteric, renal) decreased by 17.1% (from 49.7 to 41.2), while the number of endovascular aortic aneurysm repair procedures increased by 298.8% (from 16.9 to 50.5). Specifically, open aortic aneurysm procedures decreased by 21.8%, aortobifemoral bypass increased by 3.2%, and open mesenteric or renal procedures decreased by 13%. Infrainguinal bypass procedures remained relatively constant (from 37.6 to 36.5, 2.9% decrease), and the number of carotid endarterectomy procedures performed did not change significantly (from 43.6 to 42.2, 3.2% decrease). CONCLUSION: Vascular surgery trainees are performing a vastly increased total number of procedures. This increase in total procedure volume is almost entirely attributable to the recent increase in endovascular procedures. Aside from a small decline in open aortic procedures, the volume of open surgical procedures has largely remained stable. It is essential that vascular surgery training programs continue to focus on both endovascular and open surgical skills in order for vascular surgeons to remain the premier specialists to care for patients with vascular disease. PMID- 19217744 TI - Endovascular revascularization of renal artery stenosis in the solitary functioning kidney. AB - BACKGROUND: Endovascular therapy for symptomatic atherosclerotic renal artery stenosis (ARAS) is considered effective. This study evaluates the factors that impact long term anatomic and functional outcomes of endovascular therapy of ARAS in patients with a solitary functioning kidney. METHODS: We performed a retrospective analysis of records from patients who underwent endovascular intervention for ARAS and identified patients with a solitary functioning kidney (absent or nonfunctioning contralateral kidney) and patients with contralateral normal kidney (for comparison) between January 1990 and January 2008. Indications for intervention in the solitary functioning kidney were poorly controlled hypertension (diastolic blood pressure [BP] >90 mm Hg on >3 antihypertensive medications) and/or elevated creatinine (Cr >/=1.5 mg/dL). Clinical benefit was defined as freedom from composite recurrent symptoms (recurrent hypertension or renal-related morbidity-increase in persistent creatinine >20% of baseline, progression to hemodialysis, and death from renal-related causes), anatomic patency and patient survival were measured. RESULTS: A total of 242 patients (56% male, average age 69 years, range, 45-90) underwent angioplasty (23%) or primary stenting (77%) of a single renal artery with a normal contralateral renal vessel and kidney and 73 patients (58% male, average age 70 years, range, 52-89) underwent angioplasty (37%) or primary stenting (63%) for a solitary functioning kidney. There were no significant differences in mortality or morbidity between the groups. There was a significant difference in the long-term survival with 55 +/- 8% patients with a normal contralateral kidney vs 27 +/- 7% patients with a solitary functioning kidney alive at 10 years. Clinical benefit was 67 +/- 6% and 67 +/- 4% at 5 years and 63 +/- 8% and 62 +/- 4% at 10 years for solitary functioning kidney and normal contralateral groups, respectively. Using proportional hazard analysis, the predictors of long-term clinical benefit were ipsilateral kidney size (>9 cm), no immediate deterioration in function, and an estimated Glomerular Filtration Rate (eGFR) >30 mL/min/1.73m(2). Neither control of diabetes nor the administration of statins was shown to influence outcomes in the solitary functioning kidney. CONCLUSION: Intervention in patients with a solitary functioning kidney is a safe procedure and improves or stabilizes renal function in 82% of patients. Clinical benefit is dictated by preoperative GFR, renal size, and the occurrence of acute functional injury after the procedure. PMID- 19217745 TI - The vascular and biochemical effects of cilostazol in patients with peripheral arterial disease. AB - OBJECTIVES: Cilostazol improves walking distance and quality of life in patients with peripheral arterial disease (PAD). This study assessed the vascular and biochemical effects of cilostazol therapy in PAD patients. METHODS: PAD patients were prospectively recruited to a randomized, double-blinded, placebo-controlled trial. Baseline clinical data were recorded. Clinical assessment included measurement of arterial compliance, transcutaneous oxygenation, ankle-brachial index (ABI), and treadmill walking distance. Blood analyses included a full blood panel, coagulation screen, urea and electrolytes, liver function tests, estimated glomerular filtration rate, and lipid profiles. Quality of life indices were recorded using validated generic and walking-specific questionnaires. All tests were performed at baseline, 6, and 24 weeks. RESULTS: Eighty patients (53 men) were recruited from December 2004 to January 2006. The cilostazol group had a significant reduction in the augmentation index compared with the placebo group at 6 weeks (19.7% vs 26.7%, P = .001) and at 24 weeks (19.7% vs 27.7%, P = .005). A paradoxic reduction in transcutaneous oxygenation levels was identified in the cilostazol group for the left foot at 6 weeks and for the right foot at both 6 and 24 weeks. The ABIs were not significantly different between treatment groups at baseline, 6 weeks, or 24 weeks for the left and right lower limbs. The mean percentage change in walking distance from baseline improved more markedly in the cilostazol compared with the placebo group for absolute claudication distance at 6 (78.6% vs 26.4%, P = .20) and 24 weeks (173.1% vs 92.1%, P = .27); however, these failed to reach significance. Significant improvements in lipid profiles were demonstrated with cilostazol therapy at 6 weeks (triglycerides, high-density lipoprotein [HDL]) and at 24 weeks (cholesterol, triglycerides, HDL, and low density lipoprotein). The cilostazol treatment group demonstrated significant improvements in the Short Form-36 (physical functioning, physical component score), Walking Impairment (distance and speed), and Vascular Quality of Life (pain) indices at 6 and 24 weeks. Although cilostazol was associated with side effects in approximately one-third of patients, most settled within 6 weeks, facilitating the continuation of therapy in >89%. CONCLUSION: Cilostazol is a well-tolerated, safe, and efficacious treatment for PAD patients. It not only improves patients' symptomatology and quality of life but also appears to have beneficial effects on arterial compliance, possibly through its lipid-lowering property. PMID- 19217746 TI - A new clinicopathological entity of IgG4-related inflammatory abdominal aortic aneurysm. AB - OBJECTIVE: Recently, the relationship between immunoglobulin (Ig)G4 and idiopathic sclerosing lesions has attracted much attention. IgG4-related disease was first described with regard to the pancreas (autoimmune pancreatitis), and has been expanded to various organ systems. We previously reported that inflammatory abdominal aortic aneurysm (IAAA) could be one of the manifestations of IgG4-related disease. In this study, we tried to elucidate the clinical characteristics of IgG4-related IAAA. METHODS: This study consisted of 23 cases of IAAA and 40 cases of atherosclerotic abdominal aortic aneurysm (AAA). Clinical presentation, laboratory findings, and pathological features were examined. Aneurysms of 13 cases histologically corresponded to IgG4-related IAAA. RESULTS: Those cases accounted for 5% of all surgical AAAs, and 57% of IAAAs. Compared to non-IgG4-related IAAA, IgG4-related cases were characterized by less frequent association with abdominal or back pain. Serum IgG4 concentrations were significantly elevated in IgG4-related cases. Interestingly, patients with IgG4 related IAAA frequently showed an allergic constitution, such as drug allergy, autoimmune diseases, high serum IgE concentrations, and a high titer of antinuclear antibody. Pathologically, IgG4-related cases were characterized by more significant thickening of the adventitia and more numerous IgG4-positive plasma cell infiltrations. Three non-IgG4-related cases showed aneurysmal rupture at the time of first presentation, whereas no IgG4-related cases showed rupture. CONCLUSION: Recognizing a new disease entity of IgG4-related IAAA seems important because this was clinically and pathologically different from conventional aAAA and non-IgG4-related IAAA. PMID- 19217747 TI - Regional variation in the utilization of carotid endarterectomy. AB - OBJECTIVE: In different regions of the United States, highly variable rates have been documented for a wide range of procedures, such as prostatectomy and caesarean section. It is generally held that this variation is due to inconsistent physician practice patterns or other nonmedical considerations. Only limited research has been conducted regarding vascular surgical operations. We examined national data on the utilization of carotid endarterectomy (CEA) to determine the extent and diversity of regional variations. METHODS: Medicare discharge data quantified the per capita rate of CEA in 50 states and the District of Columbia in 2003. Multiple metrics pertaining to risk factors, socioeconomic status, access to care, provider density, and local health care capacity were quantified. We performed bivariate analysis, Pearson (PC) or Spearman (SC) correlations, and multiple regression modeling. RESULTS: In 2003, 83,164 CEAs were performed on 28,767,985 enrollees. CEA rates were 28.9 +/- 7.8 per 10,000 (range, 5.6-44.7 per 10,000). The rate of CEA was highly correlated with the number of heart disease deaths (PC = 0.575, P < .0001), deaths by stroke (PC = 0.504, P = .0002), and percentage of adult smokers in a state (PC = 0.643, P < .0001). These three factors held the strongest association with variation in CEA rates. Statistically, they explained 51% of the variation in total number of CEAs (R(2) = 0.5074, P < .0001). Median annual income (PC = -0.608, P < .0001) and percentage of college degrees (PC = -0.606, P < .0001) displayed inverse relationships to CEA rates. Per capita hospital beds (SC = 0.540, P < .0001) and rural health care clinics (SC = 0.518, P < .0001) exhibited positive correlations. The number of physicians or vascular surgeons did not predict higher utilization of CEA. CONCLUSION: The strongest correlations for CEA were three markers associated with atherosclerotic disease: percentage of adult smokers and deaths from heart disease or stroke. Geographic variation in this vascular procedure is chiefly associated with variance in markers of disease prevalence, not physician preference or other nonmedical factors. The increased utilization of carotid stenting, accompanied by the participation of a much wider range of medical specialists, may affect this relationship in the future. PMID- 19217748 TI - Outcomes of endoluminal reintervention for restenosis after percutaneous renal angioplasty and stenting. AB - BACKGROUND: Endovascular therapy for symptomatic atherosclerotic renal artery stenosis (ARAS) is common but is associated with a significant restenosis rate. This study evaluates the outcomes of percutaneous endoluminal therapy for symptomatic restenosis (>50% on angiography) after renal intervention. METHODS: We performed a retrospective analysis of records from patients who underwent endovascular intervention for ARAS and were followed by duplex ultrasound scan between January 1990 and January 2008. Clinical benefit defined as freedom from recurrent hypertension or renal-related morbidity (increase in persistent creatinine >20% of baseline, progression to hemodialysis, death from renal related causes), anatomic patency, restenosis, and patient survival were measured. RESULTS: A total of 447 patients underwent 619 renal artery interventions. A total of 80 vessels restenosed with an actuarial restenosis rate of 19% at 5 years. Of these restenoses, 65 (81%) were associated with recurrent symptoms (recurrent hypertension 84%, or continuing deterioration in renal function 16%). Fifty-five (85%) underwent repeat angioplasty and 10 underwent bypass surgery. The remainder was observed. The 55 percutaneous interventions were performed in 51 patients (61% female, average age 62 years, range, 51-85). A total of 73% had metabolic syndrome, 58% had hyperlipidemia, and 51% were considered diabetic; all of them had primary stenting during their first procedure. There was a 4% technical failure rate in both groups. In the restenosis group, the presence of stent was associated with a 9% technical failure rate, while in the absence of a stent the technical failure rate was only 3% (P < .05). At 5 years, outcomes were equivalent between the primary and recurrent groups for survival (76 +/- 2% vs 75 +/- 8%, primary vs recurrent), cumulative patency (82 +/- 3% vs 70 +/- 10%), freedom from restenosis (81 +/- 3% vs 81 +/- 9%), and retained clinical benefit (44 +/- 4% vs 46 +/- 10%). By Cox proportional hazards and multivariate analysis, administration of statins were associated with freedom from restenosis in the recurrent lesions. Statins, contralateral kidney size (>9 cm) and a >/=20% improvement in baseline creatinine with 3 months were associated with freedom from recurrent symptoms. Restenosis after therapy in recurrent lesions was significantly correlated with recurrent symptoms (Spearman r = 0.4614, P < .0004). CONCLUSION: Percutaneous reintervention for renal artery restenosis is safe and effective with equivalent outcomes to primary intervention. The patients are more likely to present with recurrent hypertension and be younger and of female gender than patients presenting for primary intervention. Functional outcomes after reintervention are equivalent to primary intervention. PMID- 19217749 TI - Gait variability is altered in patients with peripheral arterial disease. AB - OBJECTIVE: Claudication is the most common presentation of peripheral arterial disease (PAD), producing significant ambulatory compromise. Claudicating patients, most of whom are elderly, have reduced mobility and poor health outcomes, including an increased risk of falls. The gait of elderly fallers is characterized by increased variability. Increase in the variability of the locomotor system makes the gait more noisy and unstable. The purpose of this study is to investigate gait variability in patients with PAD. METHODS: Nineteen symptomatic PAD patients (age, 63.6 +/- 9.8 years; body mass, 82.1 +/- 18.5 kg; height, 1.71 +/- 0.06 m) walked on a treadmill in the absence of pain or claudication symptoms while joint flexion and extension kinematics were captured. Results were compared with results obtained from 17 matched healthy controls (age, 65.2 +/- 12.5 years; body mass, 82.0 +/- 25.9.5 kg; height, 1.73 +/- 0.08 m). Relative joint angles were calculated for the ankle, knee, and hip flexion/extension, and the stride-to-stride variability of joint flexion and extension was calculated from at least 30 consecutive footfalls. Variability was expressed using the largest Lyapunov exponent, standard deviation, and coefficient of variation. Independent t tests were used to compare gait variability between groups. RESULTS: Symptomatic PAD patients had significantly higher largest Lyapunov exponent values and coefficient of variation values for all joints, and higher standard deviation values at the ankle and the hip (P < .05). CONCLUSION: Symptomatic PAD patients have increased gait variability at the ankle, knee, and hip joints at baseline ambulation in the absence of claudication pain. Our findings indicate significant baseline deterioration in the locomotor system of symptomatic PAD patients. This deterioration results in increased noise and instability of gait and is a potential contributing factor to the falls and mobility problems experienced by symptomatic PAD patients. PMID- 19217750 TI - Thrombin promotes arteriogenesis and hemodynamic recovery in a rabbit hindlimb ischemia model. AB - BACKGROUND: Compared with angiogenesis, arteriogenesis is a distinct process based on the remodeling and maturation of pre-existing arterioles into large conductance arteries. Therapeutic angiogenesis has been proposed as a potential treatment for ischemic atherosclerotic diseases. Since a variety of angiogenic factors have been tested with inconsistent so far clinical results, the challenge remains in identifying the factor(s) that will stimulate functional neovascularization. Thrombin has been reported to play a pivotal role in the initiation of angiogenesis by regulating and organizing a network of angiogenic mediators. Also, it was recently demonstrated that thrombin is a potent anti apoptotic factor for endothelial cells, providing evidence on a potential role of thrombin in vascular protection and maintenance of vessel integrity. Based on these observations, we hypothesized that thrombin may promote the development of mature functional blood vessels. METHODS: Seventy-four (n=74) rabbits underwent bilateral femoral artery surgical excision. On the 20th postsurgical day increasing doses of VEGF or bFGF or thrombin were injected in one ischemic limb per rabbit and an equal volume of normal saline to the contralateral control limbs. Quantification of newly developed collateral vessels (diameter >500 mum) was performed by transauricular intra-arterial subtraction angiography. Computerized quantitative analysis of collateral vessels in angiography images was based on the concept of multiscale structural tensor. Perfusion analysis of an in vivo dynamic computed tomography study was performed to investigate hemodynamic recovery of the distal ischemic limbs. Tissue perfusion analysis was performed with the semiquantitative slope methodology, which focuses on the first pass arterial phase. RESULTS: A single administration of thrombin exhibited a dose-dependent increase of arteriogenic outcome. Thrombin at 5000 IU induced a 30.2 +/- 7.4% (P < 0.05) increase of total collateral area and length. Both VEGF and bFGF were without any significant effect at the concentrations used. Functional estimation of limb perfusion showed a statistically significant increase of blood flow recovery only for thrombin. The semiquantitative slope method perfusion score differed significantly in the 5000 IU thrombin treated limbs (5.7 +/- 0.3 vs 5.0 +/- 0.3 in control ischemic limbs; P < .05), and was not significantly inferior from the score of normal nonoperated limbs (6.5 +/- 0.3) suggesting a trend towards hemodynamic recovery of distal limb perfusion. CONCLUSIONS: In a rabbit hindlimb ischemia model, thrombin promoted the formation of large collateral vessels and improved the perfusion of distal ischemic tissue. These results provide new insights in understanding the involvement of thrombin in vascular formation and point to a novel role of thrombin in arteriogenesis. PMID- 19217751 TI - [Renal artery stenosis : atheromatous disease and fibromuscular dysplasia]. AB - Renal artery stenosis may be due to atheromatous disease or renal fibromuscular dysplasia (FMD). Management of both diseases requires treatment of hypertension usually observed in such patients; however, clinical presentation, mechanism and treatment of these 2 diseases are usually different. Renal FMD is now considered as a systemic disease, the cause of which may be genetic (although the exact cause is still elusive). Renal arteries are the most frequent localizations of FMD, but extra renal arteries may also be involved (usually carotid arteries). Risk factors of hypertension-induced renal FMD include estrogen treatment and smoking. Renal FMD are mostly found in young women and in children who present with recent severe and/or refractory symptomatic hypertension. Diagnosis is usually easy (Doppler, CT-scan), and treatment of renal FMD is angioplasty in most cases. Atheromatous renal artery stenosis is usually found in patients with other atheromatous disease (peripheral artery disease, carotid, coronary artery disease...). Clinical presentation include severe or refractory hypertension, recurrent flash pulmonary edema in a patient with hypertension, progressive renal dysfunction spontaneously or after medical treatment with converting-enzyme inhibition or angiotensin II blockade, hypertension in a patient (usually smoker or ex-smoker) with diffuse atheromatous vascular disease. Management of atheromatous renal artery disease is medical treatment in all patients (aggressive treatment of cardiovascular risk factors, control of arterial pressure); revascularization is required in some patients only since it rarely cures hypertension: the goal of revascularization is mostly renal function protection, which may be observed in selected patients. Revascularization must be decided by physicians or teams involved in the care of such patients. Patients with atheromatous renal artery disease are at very high renal and cardiovascular risk : aggressive management of cardiovascular risk factors is crucial. PMID- 19217752 TI - Unguided sacroiliac injection: effect on refractory buttock pain in patients with spondyloarthropathies. AB - OBJECTIVE: This study was designed to evaluate the efficacy and safety of unguided sacroiliac injections for refractory sacroiliac pain due to spondyloarthropathies. METHOD: An open-label clinical trial lasted 20 weeks for each of 29 patients, with different subtypes of spondyloarthropathies. It was conducted from September 2004 through January 2007. In patients with refractory inflammatory buttock pain (the inclusion criterion), we performed unguided injections of triamcinolone acetonide 40 mg into each sacroiliac joint on an outpatient basis. Assessments by patients and clinicians were recorded at baseline and every 4 weeks until the end of the study. RESULTS: At week 4, significant decreases were seen in the patients' assessment of pain, morning stiffness, and sleep disturbance and in the clinicians' sacroiliac examination score. Pain reduction, as scored by patient and clinician, was maintained through week 20. No significant improvement was seen on the finger-to-floor or Schober tests. Magnetic resonance imaging (MRI) results showed a significant decrease in inflammatory scores after the sacroiliac injections. The patients' pain score and sleep disturbance were both clearly associated with the MRI inflammation index. The injection was well tolerated by all patients and no serious adverse event was seen. CONCLUSION: This technique for sacroiliac injection was effective, and the final results showed a significant improvement in pain, similar to that with imaging-guided interventions. Trained physicians can perform this procedure safely in outpatient settings. PMID- 19217753 TI - Substance abuse, coping strategies, adaptive skills and behavioral and emotional problems in clients with mild to borderline intellectual disability admitted to a treatment facility: a pilot study. AB - Many clients with mild to borderline intellectual disability (ID) who are admitted to a treatment facility show serious problems in alcohol and/or drugs use. In the present case file study, we explored differences in coping strategies, adaptive skills and emotional and behavioral problems between clients who showed substance abuse and clients who did not. There were no differences in adaptive skills between groups. However, compared to clients without substance abuse, those who abused substances showed a more palliative coping style, and had more severe emotional and behavior problems such as anxiety/depression and intrusive thoughts and aggressive and antisocial behaviors. Implications for treatment are discussed. PMID- 19217754 TI - Evaluation of uterine cervix segmentations using ground truth from multiple experts. AB - This work is focused on the generation and utilization of a reliable ground truth (GT) segmentation for a large medical repository of digital cervicographic images (cervigrams) collected by the National Cancer Institute (NCI). NCI invited twenty experts to manually segment a set of 939 cervigrams into regions of medical and anatomical interest. Based on this unique data, the objectives of the current work are to: (1) Automatically generate a multi-expert GT segmentation map; (2) Use the GT map to automatically assess the complexity of a given segmentation task; (3) Use the GT map to evaluate the performance of an automated segmentation algorithm. The multi-expert GT map is generated via the STAPLE (Simultaneous Truth and Performance Level Estimation) algorithm, which is a well-known method to generate a GT segmentation from multiple observations. A new measure of segmentation complexity, which relies on the inter-observer variability within the GT map, is defined. This measure is used to identify images that were found difficult to segment by the experts and to compare the complexity of different segmentation tasks. An accuracy measure, which evaluates the performance of automated segmentation algorithms is presented. Two algorithms for cervix boundary detection are compared using the proposed accuracy measure. The measure is shown to reflect the actual segmentation quality achieved by the algorithms. The methods and conclusions presented in this work are general and can be applied to different images and segmentation tasks. Here they are applied to the cervigram database including a thorough analysis of the available data. PMID- 19217755 TI - Heterotopic neuroglial tissue as a congenital laterocervical mass: a case report. AB - Heterotopic neuroglial tissue in the head and neck area is a rare clinical entity which can cause airway obstruction and feeding problems during the neonatal period. The case is presented of heterotopic neuroglial tissue as a congenital laterocervical and intraoral mass in a neonate. PMID- 19217756 TI - Genetic study of BDNF, DRD3, and their interaction in tardive dyskinesia. AB - Tardive dyskinesia (TD) is a neuroleptic-induced movement disorder. Its pathophysiology is unclear. The most consistent genetic findings have shown an association with the Ser9Gly polymorphism of the DRD3 gene. However, only few polymorphisms within DRD3 has been tested, and a comprehensive examination of DRD3 in TD is still lacking. Further, brain-derived neurotrophic factor (BDNF), a neuronal growth and survival peptide, regulates DRD3 expression and may be involved in the neuronal degeneration observed in TD. In the present study, we investigated 15 tag DRD3 polymorphisms and four tag BDNF polymorphisms for association with TD in our sample of Caucasian schizophrenia patients (N=171). While BDNF markers showed no association, a haplotype containing rs3732782, rs905568, and rs7620754 in the 5' region of DRD3 was associated with TD diagnosis (p[10,000 permutations]=0.007). We also found evidence of interaction between BDNF and DRD3 polymorphisms. Further studies are needed to confirm these findings. PMID- 19217757 TI - A voxel-based diffusion tensor imaging study of temporal white matter in patients with schizophrenia. AB - Diffusion tensor imaging (DTI) is a relatively new technique used to detect changes in the anisotropic diffusion of white matter. The study of the disruption of brain connectivity may increase our understanding of cognitive deficits associated with schizophrenia. Here we analysed DTI data in 25 patients with DSM IV schizophrenia and 24 healthy controls. Two complementary measures, fractional anisotropy (FA) and the apparent diffusion coefficient (ADC), were considered and analysed using voxel-based morphometry. Declarative memory functions were also investigated and their associations with DTI data were analysed. FA was significantly reduced, and the ADC increased in the left sub-gyral white matter of the temporal lobe, which involves the posterior part of the fornix. In the schizophrenic group, females had lower FA than males in the genu of the corpus callosum. Memory functions correlate with FA values. These data provide further evidence for the disruption of white matter connectivity in the left medial temporal lobe, and for its contribution to the declarative memory deficit in schizophrenia. PMID- 19217758 TI - Medial prefrontal default-mode hypoactivity affecting trait physical anhedonia in schizophrenia. AB - Anhedonia, as a deficit symptom, may be associated with default-mode hypofrontality in schizophrenia. To explore whether trait anhedonia in schizophrenia pertains to altered hypofrontal resting state brain function, resting state metabolic activities were compared and correlated with the Physical and Social Anhedonia Scale scores in 29 patients with schizophrenia and 21 healthy controls using (18)F-fluorodeoxyglucose positron emission tomography ((18)F-FDG-PET). In patients with schizophrenia, Physical Anhedonia Scale scores showed a trend-level negative correlation with the hypoactive dorsomedial prefrontal metabolism while Positive and Negative Symptom Scale (PANSS) negative subscale scores positively correlated with hyperactive cerebellar metabolism. Voxelwise correlation analysis showed physical anhedonia correlates in resting state activities of the supplementary motor area, ventromedial and dorsolateral prefrontal cortex, insular gyrus, and the precuneus in patients with schizophrenia while no frontal metabolic correlates were found in healthy controls. The hypoactive dorsomedial prefrontal metabolism correlated with physical anhedonia-correlated resting state regional activities. These findings provide further evidence for the relation of functional hypofrontality to the deficit syndrome and possible involvement of the functional imbalance in the cortico-cerebellar-thalamic-cortical circuit in the resting state brain function of schizophrenia. PMID- 19217760 TI - Kinetics of solid-state NMR cross-polarization from protons to carbon-13 in surgical sutures. AB - Commercial Dexon surgical sutures, made of polyglycolide (PGA), were examined using (13)C CP/MAS NMR. The study shows that detailed analysis of the cross polarization (CP) process is useful in the peak assignments and in the assessment of molecular mobility in the polymer domains. Crystallinity of PGA in the sutures was estimated at ca. 55%. PMID- 19217759 TI - Accelerated hippocampal atrophy rates in stable and progressive amnestic mild cognitive impairment. AB - Studies suggest that smaller hippocampal volume predicts Alzheimer's disease (AD) in mild cognitive impairment (MCI). However, few studies have demonstrated decline rates in cognition and hippocampal volume in MCI subjects with stable clinical presentation. Furthermore, the effects of apolipoprotein E (ApoE) on the change rates of medial temporal structures and cognition in MCI are rarely investigated. Fifty-eight subjects with amnestic MCI and 20 normal aging elderly controls received annual neuropsychological and magnetic resonance imaging (MRI) assessments. Annual decline rates in neuropsychological test scores, hippocampal and amygdalar volumes were calculated. ApoE genotypes were examined. Nineteen (32.7%) MCI subjects converted to AD during an average 22.5-month follow-up period. The annual hippocampal atrophy rate was correlated with a decline in memory test scores. The presence of the ApoE varepsilon4 allele did not affect the change rates in neuropsychological test scores and medial temporal structures volume. Compared to subjects with stable MCI (MCI-S) and normal aging, progressive MCI (MCI-P) had the highest annual decline rates in cognition and hippocampal volume. Logistic regression analysis showed that higher annual decline rates in hippocampal volume and global cognitive test scores were associated with conversion to AD. Furthermore, although MCI-S subjects had little cognitive decline, their hippocampal atrophy rates were higher than those of normal aging controls. Therefore, accelerated hippocampal atrophy rates may be an early and important presentation in MCI subjects. PMID- 19217761 TI - [Long QT syndrome: an underestimated cause of sudden infant death]. AB - Congenital long QT syndrome (LQTS) is an inherited arrhythmia that can be sporadic or familial. It predisposes to sudden cardiac death by ventricular fibrillation, which can occur at any age, particularly in neonates. Recent postmortem molecular screening surveys have shown that 10 to 12% of sudden infant death syndrome (SIDS) cases were potentially related to congenital long QT syndrome. Current SIDS etiological surveys fail to diagnose LQTS. Specific questioning and electrocardiographic screening of first-degree relatives could greatly facilitate LQTS diagnosis. We propose adding these to screening modalities after a SIDS incident. Neonatal electrocardiographic screening could allow early identification of LQTS and adapted treatment and follow-up. PMID- 19217762 TI - Fatty acid uptake by breast cancer cells (MDA-MB-231): effects of insulin, leptin, adiponectin, and TNFalpha. AB - In order to exert metabolic effects, fatty acids must be taken up by cells and metabolize effectively to different classes of cellular lipids (triacylglycerols, phospholipids, etc.) for incorporation into different cellular and intracellular compartments. Therefore, the main aim of the present study is to investigate the uptake and metabolism of fatty acids representing three different series of fatty acids such as oleic acid, 18:1n-9 (OA), arachidonic acid, 20:4n-6 (AA), and eicosapentaneoic acid, 20:5n-3 (EPA) by breast cancer cells, MDA-MB-231. Moreover, we investigated the effects of insulin and several adipokines on the fatty acid uptake by these cells as obesity and insulin resistance syndrome have been suggested to affect breast cancer risk. We report for the first time that AA was predominantly taken up by these cells compared with EPA and OA. Pre incubation of these cells with TNFalpha stimulated most of the uptake of EPA (30%), whereas uptake of OA and AA was stimulated only 10-15% compared with the controls. Insulin, leptin, and adiponectin had no effect on fatty acid uptake by these cells. Together these results demonstrate that preferential uptake of AA in MDA-MB-231 cells, and the fatty acid uptake activity of these cells is influenced by TNFalpha. PMID- 19217763 TI - Docosahexaenoic acid is a substrate for ACAT1 and inhibits cholesteryl ester formation from oleic acid in MCF-10A cells. AB - MCF-10A breast epithelial cells treated with docosahexaenoic acid (DHA) or oleic acid (OA) accumulated cytoplasmic lipid droplets containing both triacylglycerol and cholesteryl esters (CE). Interestingly, total CE mass was reduced in cells treated with DHA compared to cells treated with OA, and the CEs were rich in n-3 fatty acids. Thus, we hypothesized that DHA may be, in addition to a substrate, an inhibitor of cholesterol esterification in MCF-10A cells. We determined that the primary isoform of acyl-CoA: cholesterol acyltransferase expressed in MCF-10A cells is ACAT1. We investigated CE formation with DHA, OA, and the combination in intact cells and isolated microsomes. In both cells and microsomes, the rate of CE formation was faster and more CE was formed with OA compared to DHA. DHA substantially reduced CE formation when given in combination with OA. These data suggest for the first time that DHA can act as a substrate for ACAT1. In the manner of a poor substrate, DHA also inhibited the activity of ACAT1, a universally expressed enzyme involved in intracellular cholesterol homeostasis, in a cell type that does not secrete lipids or express ACAT2. PMID- 19217764 TI - Personality, adherence, asthma control and health-related quality of life in young adult asthmatics. AB - BACKGROUND: Striving for improved adherence and asthma control is of vital concern in today's asthma management. Several influential factors have been identified, but the importance of personality traits has been insufficiently explored. The aim was first to determine whether personality traits in young adult asthmatics are related to asthma control and health-related quality of life (HRQL), and second to examine the influences of personality traits on adherence to regular asthma medication treatment. METHODS: Young adult asthmatics, 22 years of age (n=268) completed questionnaires. Statistical analyses were performed. RESULTS: The personality traits Negative Affectivity and Impulsivity correlated negatively with asthma control, whereas in women Hedonic Capacity correlated positively with asthma control. Negative Affectivity, Impulsivity, Hedonic Capacity, Alexithymia and asthma control predicted the mental dimension of HRQL. Asthma control and physical activity predicted the physical dimension of HRQL. Among respondents with regular asthma medication (n=109), Impulsivity correlated negatively with adherence. In men, Antagonism and Alexithymia were associated with low adherence. Additionally, Alexithymia, Hedonic Capacity and Negative Affectivity showed non-linear relationships with adherence, meaning that initially increased scores on these personality traits scales were associated with increased adherence but higher scores did not increase adherence. Respondents who were prescribed a single inhaler combining ICS and LABA reported higher adherence than those with monotherapies. CONCLUSION: These data suggest that personality can influence how asthma patients adhere to asthma medication treatment, and report their control and HRQL. Tools determining personality traits may be useful in the future in individualizing management of asthma patients. PMID- 19217765 TI - Pulmonary rehabilitation is successful for COPD irrespective of MRC dyspnoea grade. AB - BACKGROUND: It is not clear whether the benefits of pulmonary rehabilitation (PR) apply equally to patients with Chronic Obstructive Pulmonary Disease (COPD) with different levels of starting disability. We have therefore investigated the effect of pulmonary rehabilitation stratified by the MRC dyspnoea scale in patients with COPD. METHODS: This is a retrospective, observational study of data collected from 450 consecutive patients with COPD attending outpatient PR: 247 male, mean (SD) age 69.5 (8.9) yrs and FEV(1) 44.6 (19.7)% predicted. The Incremental Shuttle Walk Test (ISWT) was performed before and after the seven week course RESULTS: 395 patients (88%) completed the programme. The mean (SD) baseline ISWT performance was 167 (113)m. The distribution of baseline MRC grades was 2 - 15.4%, 3 - 24.9%, 4 - 27.3% and 5 - 32.4%. The mean (95% CI) improvement in ISWT after PR for each MRC scale grade was highly significant (p<0.0005); 2 - 66 (50-83)m, 3 - 63 (50-75)m, 4 - 59 (49-70)m, and 5 - 54 (43-64)m. CONCLUSIONS: Patients with COPD, of all MRC dyspnoea grades, benefit comparably from pulmonary rehabilitation achieving both statistically and clinically meaningful improvements in exercise performance. MRC grade should therefore not be used to exclude patients from pulmonary rehabilitation. PMID- 19217766 TI - Translating biology into clinic: the case of glioblastoma. AB - GBM, the most common and malignant primary tumor of the CNS, is characterized by exponential growth and diffuse invasiveness. Although the diverse causative genotypes that give rise to a inhomogeneous histological phenotype are well defined, effective therapy inducing tumor cell apoptosis has not been established so far. Following surgery, billions of invasive tumor cells remain to be targeted by systemic and local therapies. Targeting non-overlapping pathways, rather than a single agent approach, is more likely to be effective. The potential of local drug application has not been exploited yet. Systemically, novel drug combinations have to be developed that not only target key molecules at the signaling crossroads but also exploit energy demand and the epigenetic cancer program of GBM. PMID- 19217767 TI - Targeting protein kinases for the development of anti-inflammatory drugs. AB - In recent years, protein kinases have become the pharmaceutical industry's most studied class of drug target, and some 10 protein kinase inhibitors have so far been approved for the treatment of cancer. However, whether safe drugs that modulate protein kinase activities can also be developed for the treatment of chronic diseases, where they may need to be taken for decades, is an issue that is still unresolved. A number of compounds that inhibit the p38alpha MAPK have entered clinical trials for the treatment of rheumatoid arthritis and psoriasis, but side effects have prevented their progression to Phase III clinical trials. Here I briefly review the potential problems in targeting p38 MAPK and discuss other protein kinases that regulate the innate immune system, such as Tpl2, MAPKAP-K2/3, MSK1/2 and IRAK4, which may be better targets for the treatment of chronic inflammatory diseases, and NIK, which is an attractive target for the treatment of multiple myeloma, a late stage B-cell malignancy. PMID- 19217768 TI - Nitrate and phosphate leaching in a Phaeozem soil treated with biosolids, composted biosolids and inorganic fertilizers. AB - The use of organic wastes in agriculture may increase the production of crops by incorporating organic matter and nutrients into the soil, and by improving its physical characteristics; however, this use may cause environmental problems such as the leaching of certain ions. The objective of this study was to establish possible nitrogen and phosphorus leaching under real field conditions in Phaeozem soils. The experimental work was performed in a corn (Zea mays L.) field where three plots were conditioned with inorganic fertilizer, three plots with 4.5 Mgha(-1) of biosolids on dry basis, and three plots with the same amount of composted biosolids. The quality of biosolids and composted biosolids complied with the Mexican Official Standards. Soil water samples were collected with suction cups during two agricultural cycles and were analysed. Soil samples were also taken and analysed. The N-NO(3) concentrations in soil water fluctuated between 0.9 and 98mgL(-1) in the composted biosolid treatment, between 0.7 and 64 mgL(-1) in the biosolid treatment, and between 1 and 61 mgL(-1) in the inorganic fertilizer treatment. The maximum concentration of N-NO(2) and N-NH(3) in soil water was 1.02 and 2.65 mgL(-1), respectively. The greatest percentage of nitrogen leached is produced when inorganic fertilizer is used (37.4% and 24.0% N leached in the first and second years, respectively), followed by composted biosolids (17.1% and 13.5% N leached in the first and second years, respectively) and last by biosolids (11% for both years). This difference could be related to the form in which nitrogen is present in the fertilizers, while commercial fertilizer is as inorganic nitrogen, organic wastes are basically presented as organic nitrogen. The maximum PO(4)(3-) concentration in soil water was 1.9 mgL( 1) in the composted biosolid treatment, 1.7mgL(-1) in the biosolid treatment and 0.9 mgL(-1) in the inorganic fertilizer treatment. The estimated percentage of leached phosphorus was less than 1% for all treatments. The minimum leaching that occurred seemed to be due to a sorption-precipitation process. PMID- 19217770 TI - Protein folding and binding: moving into unchartered territory. PMID- 19217769 TI - Selection of imprinted nanoparticles by affinity chromatography. AB - Soluble molecularly imprinted nanoparticles were synthesised via iniferter initiated polymerisation and separated by size via gel permeation chromatography. Subsequent fractionation of these particles by affinity chromatography allowed the separation of high affinity fractions from the mixture of nanoparticles. Fractions selected this way possess affinity similar to that of natural antibodies (K(d) 6.6x10(-8)) M and were also able to discriminate between related functional analogues of the template. PMID- 19217772 TI - The future of anaerobic digestion and biogas utilization. AB - One of the common tendencies of animal production activities in Europe and in developed countries in general is to intensify the animal production and to increase the size of the animal production units. High livestock density is always accompanied by production of a surplus of animal manure, representing a considerable pollution threat for the environment in these areas. Avoiding over fertilization is not only important for environmental protection reasons but also for economical reasons. Intensive animal production areas need therefore suitable manure management, aiming to export and to redistribute the excess of nutrients from manure and to optimize their recycling. Anaerobic digestion of animal manure and slurries offers several benefits by improving their fertilizer qualities, reducing odors and pathogens and producing a renewable fuel - the biogas. The EU policies concerning renewable energy systems (RES) have set forward a fixed goal of supplying 20% of the European energy demands from RES by year 2020. A major part of the renewable energy will originate from European farming and forestry. At least 25% of all bioenergy in the future can originate from biogas, produced from wet organic materials such as: animal manure, whole crop silages, wet food and feed wastes, etc. PMID- 19217773 TI - Remediation of chromite ore processing residue by pyrolysis process with sewage sludge. AB - The present work developed a novel technique to treat chromite ore processing residue (COPR). The process involved mixing the COPR with sewage sludge followed by pyrolysis. The gaseous organic fraction generated during pyrolysis of sludge was beneficial to Cr(VI) reduction. Process variables, such as the amount of sludge added to COPR (sludge-to-COPR (S/C) ratio), heating temperature, reaction time and particle size, were systematically varied, and their influences on the Cr(VI) reduction in COPR were investigated. Cr(VI) content had decreased greatly, from 3384 mg kg(-1) for untreated COPR to less than 30 mg kg(-1) for COPR treated at 600 degrees C. PMID- 19217774 TI - Variation of ADM1 by using temperature-phased anaerobic digestion (TPAD) operation. AB - The objective of the study was to examine the application of the Anaerobic Digestion Model No. 1 (ADM1) developed by the IWA task group for mathematical modelling of anaerobic process. Lab-scale temperature-phased anaerobic digestion (TPAD) process were operated continuously, and were fed with co-substrate composed of dog food and flour. The model platform implemented in the simulation was a derivative of the ADM1. Sensitivity analysis showed that k(m.process) (maximum specific uptake rate) and K(S.process) (half saturation value) had high sensitivities to model components. Important parameters including maximum uptake rate for propionate utilisers (k(m.pro)) and half saturation constant for acetate utilisers (K(S.ac)) in the thermophilic digester and maximum uptake rate for acetate utilisers (k(m.ac)) in the mesophilic digester were estimated using iterative methods, which optimized the parameters with experimental results. Simulation with estimated parameters showed good agreement with experimental results in the case of methane production, uptake of acetate, soluble chemical oxygen demand (SCOD) and total chemical oxygen demand (TCOD). Under these conditions, the model predicted reasonably well the dynamic behavior of the TPAD process for verifying the model. PMID- 19217775 TI - Preparation and characterization of glycerol plasticized-pea starch/ZnO carboxymethylcellulose sodium nanocomposites. AB - Among natural polymers, starch is one of the most promising biodegradable materials because it is a renewable bioresource that is universally available and of low cost. However, the properties of starch-based materials are not satisfactory. One approach is the use of nano-filler as reinforcement for starch based materials. In this paper, a nanocomposite is prepared using ZnO nanoparticles stabilized by carboxymethylcellulose sodium (CMC) as the filler in glycerol plasticized-pea starch (GPS) matrix by the casting process. According to the characterization of ZnO-CMC particles with Fourier transform infrared (FTIR), Ultraviolet-visible (UV-vis), X-ray diffraction (XRD), transmission electron microscope (TEM) and thermogravimetric analysis (TG), ZnO (about 60 wt%) is encapsulated with CMC (about 40 wt%) in ZnO-CMC particles with the size of about 30-40 nm. A low loading of ZnO-CMC particles can obviously improve the pasting viscosity, storage modulus, the glass transition temperature and UV absorbance of GPS/ZnO-CMC nanocomposites. When the ZnO-CMC contents vary from 0 to 5 wt%, the tensile yield strength increase from 3.94 MPa to 9.81 MPa, while the elongation at break reduce from 42.2% to 25.8%. The water vapor permeability decrease from 4.76 x 10(-10) to 1.65 x 10(-10) g m(-1) s(-1) Pa(-1). PMID- 19217776 TI - Oxygen transfer and energy dissipation rate in surface aerator. AB - The dissipation rate of turbulent kinetic energy (epsilon) is a key parameter for mixing in surface aerators. In particular, determination epsilon across the impeller stream, where the most intensive mixing takes place, is essential to ascertain that an appropriate degree of mixing is achieved. Present work by using commercial software VisiMix calculates the energy dissipation rate in geometrically similar unbaffled surface aeration systems in order to scale-up the oxygen transfer process. It is found that in geometrically similar system, oxygen transfer rate is uniquely correlated with dissipation rate of energy. Simulation or scale-up equation governing oxygen transfer rate and dissipation rate of energy has been developed in the present work. PMID- 19217771 TI - Comparative cancer survival information in Europe. PMID- 19217777 TI - Synthesis, binding affinity and SAR of new benzolactam derivatives as dopamine D3 receptor ligands. AB - A series of new benzolactam derivatives was synthesized and the derivatives were evaluated for their affinities at the dopamine D(1), D(2), and D(3) receptors. Some of these compounds showed high D(2) and/or D(3) affinity and selectivity over the D(1) receptor. The SAR study of these compounds revealed structural characteristics that decisively influenced their D(2) and D(3) affinities. Structural models of the complexes between some of the most representative compounds of this series and the D(2) and D(3) receptors were obtained with the aim of rationalizing the observed experimental results. Moreover, selected compounds showed moderate binding affinity on 5-HT(2A) which could contribute to reducing the occurrence of extrapyramidal side effects as potential antipsychotics. PMID- 19217778 TI - [18F]FEAC and [18F]FEDAC: Two novel positron emission tomography ligands for peripheral-type benzodiazepine receptor in the brain. AB - [(18)F]FEAC ([(18)F]4a) and [(18)F]FEDAC ([(18)F]4b) were developed as two novel positron emission tomography (PET) ligands for peripheral-type benzodiazepine receptor (PBR). [(18)F]4a and [(18)F]4b were synthesized by fluoroethylation of precursors 8a and 8b with [(18)F]FCH(2)CH(2)Br ([(18)F]9), respectively. Small animal PET scan for a neuroinflammatory rat model showed that the two radioligands had high uptakes of radioactivity in the kainic acid-infused striatum, a brain region where PBR density was increased. PMID- 19217779 TI - Discovery and optimization of piperidyl benzamide derivatives as a novel class of 11beta-HSD1 inhibitors. AB - Discovery and optimization of a piperidyl benzamide series of 11beta-HSD1 inhibitors is described. This series was derived from a cyclohexyl benzamide lead structures to address PXR selectivity, high non-specific protein binding, poor solubility, limited in vivo exposure, and in vitro cytotoxicity issues observed with the cyclohexyl benzamide structures. These efforts led to the discovery of piperidyl benzamide 15 which features improved properties over the cyclohexyl benzamide derivatives. PMID- 19217780 TI - Lithospermic acid derivatives from Lithospermum erythrorhizon increased expression of serine palmitoyltransferase in human HaCaT cells. AB - A MeOH extract of the dry root of Lithospermum erythrorhizon showed strong increasing effect on serine palmitoyltransferase (SPT) in normal human keratinocyte cells (HaCaT cells). Bioassay-guided separation on this extract using repeated chromatography resulted in the isolation of lithospermic acid (1) and two derivative esters, 9''-methyl lithospermate (2) and 9'-methyl lithospermate (3). Compounds 1-3 significantly increased SPT expressions in the relative quantity (%) of SPT1 mRNA as well as SPT2 mRNA. These constituents also raised the level of SPT protein in HaCaT cells in a dose-dependent manner, with the increased level of SPT protein in HaCaT cells of 55%, 23%, and 81% at the concentration of 100 microg/ml, respectively. This finding suggests that lithospermic acid and its derivatives from L. erythrorhizon might improve the permeability barrier by stimulating the protein level of SPT. PMID- 19217781 TI - The use of oxadiazole and triazole substituted naphthyridines as HIV-1 integrase inhibitors. Part 1: Establishing the pharmacophore. AB - A series of HIV-1 integrase inhibitors containing a novel metal binding motif consisting of the 8-hydroxy-1,6-naphthyridine core and either an oxadiazole or triazole has been identified. The design of the key structural components was based on a two-metal coordination pharmacophore. This report presents initial structure-activity data that shows the new chelation architecture delivers potent inhibition in both enzymatic and antiviral assays. PMID- 19217783 TI - Ubiquitin-mediated regulation of apoptosis. AB - Ubiquitin is a protein modifier that is conjugated to target proteins either as a single moiety or as polyubiquitin chains. Over the past several years, an increasing number of ubiquitin ligases and ubiquitin-deconjugating enzymes have been identified; these modulate cell survival by degradative and non-degradative means. Mutations that affect ubiquitin-mediated signalling are tightly linked to various human pathologies including tumorigenesis. Unravelling how the ubiquitin signal is conjugated, edited and 'read' is crucial to understanding cellular processes such as endocytic trafficking, NF-kappaB signalling, gene expression, DNA repair and apoptosis. In this review, we summarize recent advances that start to elucidate how the ubiquitin message is used as a versatile tool to regulate apoptosis, for example in the conjugation of ubiquitin to caspases. This results in steric interference with substrate entry and allosteric conformational impairment of the catalytic pocket of the caspase. PMID- 19217782 TI - Identification of a selective thieno[2,3-c]pyridine inhibitor of COT kinase and TNF-alpha production. AB - COT (Tpl2 in mice) is a serine/threonine MAP3 kinase that regulates production of TNF-alpha and other pro-inflammatory cytokines such as IL-1beta via the ERK/MAP kinase pathway. As TNF-alpha and IL-1beta are clinically validated targets for therapeutic intervention in rheumatoid arthritis (RA), blocking COT provides a potential avenue for amelioration of disease. Herein we describe identification of a cellular active selective small molecule inhibitor of COT kinase. PMID- 19217784 TI - Weight distribution influences the time required to lift the leg even under normal standing condition. AB - The purpose of the present study is to demonstrate whether the time taken to initiate a lift-one-leg task from a quiet standing position is determined by the initial weight distribution (WD), irrespective of whether the initial posture is controlled spontaneously or by using visual feedback. Seven healthy young subjects performed a lift-one-leg task in response to visual stimulation provided by an LED. The experiment included the normal (N) condition in which the participants stand on both legs symmetrically, as subjectively estimated, and the three feedback (FL, FC and FR) conditions in which the participants regulate the WD at -20% (left-weighted by 6:4), 0% (center-weighted by 5:5) and 20% (right weighted by 4:6), respectively, through the use of visual feedback. In the N condition, the inter-trial variation of WD was larger than that of the FC condition, which was enough to significantly correlate with the time required to lift the leg. The slope, intercept, and standard error of estimation of the regression equation between the WD and the duration of anticipatory postural adjustment were consistent between conditions. The quantification of the effect of WD on anticipatory postural adjustment indicated that the time taken to lift the leg was delayed by 2.81ms as the center of mass shifted toward the lifting leg by 1mm. Our results demonstrate that asymmetric loading is a simple biomechanical factor that determines the time needed to initiate a subsequent lift-one-leg task. PMID- 19217785 TI - Plantar pressure relief in the diabetic foot using forefoot offloading shoes. AB - PURPOSE: Forefoot offloading shoes (FOS) are commonly used in clinical practice for treatment of plantar forefoot ulcers in the diabetic foot. The aim of this study was to assess the offloading efficacy of four different FOS models in comparison with a cast shoe and control shoe. METHODS: In-shoe plantar pressures were measured during walking in each of the six footwear conditions in 24 neuropathic diabetic patients at high risk for plantar foot ulceration. For each of six foot regions, peak pressure, pressure-time integral, and force-time integral were calculated. Load transfer diagrams were developed to assess the footwear mechanisms of action. Perceived walking comfort was measured using a visual analogue scale (VAS). All comparisons between conditions were tested at P<0.05. RESULTS: Peak pressures and pressure-time integrals at the metatarsal heads and hallux regions were significantly reduced (by 38-58%) in all FOS models when compared with the control shoe. The FOS also relieved metatarsal head peak pressure to a significantly larger extent than the cast shoe (approximately 20%). The load transfer diagrams showed a major transfer of approximately 40% of forefoot load to the midfoot explaining the offloading efficacy of the FOS. Perceived walking comfort was significantly lower in the FOS (VAS score 2.7-5.9) when compared with the control shoe (VAS 8.2) and cast shoe (VAS 6.8). CONCLUSIONS: The data showed that all FOS models were effective in their primary goal, relieving forefoot pressure in at-risk neuropathic diabetic patients. Therefore, these shoes may be effective in offloading and healing plantar forefoot ulcers, although the low comfort scores should be considered as this may potentially affect adherence to treatment. PMID- 19217786 TI - Transient upbeat nystagmus due to unilateral focal pontine infarction. AB - Upbeat nystagmus is known to be caused by an imbalance of the vertical vestibulo ocular reflex (VOR), favoring downward VOR activity, due to bilateral lesions of the medulla, ventral tegmentum, anterior cerebellar vermis, adjacent brachium conjunctivum and the midbrain. We report on two patients who had transient upbeat nystagmus due to unilateral pontine infarction that may have disrupted bilateral upward VOR pathways running in the ventral tegmental tracts. PMID- 19217787 TI - Design, synthesis, and biological evaluation of novel quinoline derivatives as HIV-1 Tat-TAR interaction inhibitors. AB - Thirty-two quinoline derivatives were designed and synthesized as HIV-1 Tat-TAR interaction inhibitors. All the compounds showed high antiviral activities in inhibiting the formation of SIV-induced syncytium in CEM174 cells. Nine of them with low cytotoxicities were evaluated by Tat dependent HIV-1 LTR-driven CAT gene expression colorimetric enzyme assay in human 293T cells, indicating effective inhibitory activities of blocking the Tat-TAR interaction. Molecular modeling experiments indicated that these compounds may inhibit Tat-TAR interaction by binding to Tat protein instead of TAR RNA. PMID- 19217789 TI - Synthesis of novel substituted pyrazole-5-carbohydrazide hydrazone derivatives and discovery of a potent apoptosis inducer in A549 lung cancer cells. AB - A series of novel 3-aryl-1-(4-tert-butylbenzyl)-1H-pyrazole-5-carbohydrazide hydrazone derivatives were synthesized and the effects of all the compounds on A549 cell growth were investigated. The results showed that all compounds had inhibitory effects on the growth of A549 lung cancer cells and compound (E)-1-(4 tert-butylbenzyl)-N'-(1-(5-chloro-2-hydroxyphenyl) ethylidene)-3-(4-chlorophenyl) 1H-pyrazole-5-carbohydrazide (3e) possessed the highest growth inhibitory effect and induced apoptosis of A549 lung cancer cells. PMID- 19217788 TI - Synthesis and bioevaluation of omega-N-amino analogs of B13. AB - Novel omega-N-amino analogs of B13 (Class E) were designed, synthesized and tested as inhibitors of acid ceramidase (ACDase) and potential anticancer agents deprived of unwanted lysosomal destabilization and ACDase proteolytic degradation properties of LCL204 [Szulc, Z. M.; Mayroo, N.; Bai, A.; Bielawski, J.; Liu, X.; Norris, J. S.; Hannun, Y. A.; Bielawska, A. Bioorg. Med. Chem. 2008, 16, 1015]. Representative analog LCL464, (1R,2R)-2-N-(12'-N,N-dimethylaminododecanoyl amino) 1-(4''-nitrophenyl)-1,3-propandiol, inhibited ACDase activity in vitro, with a similar potency as B13 but higher than LCL204. LCL464 caused an early inhibition of this enzyme at a cellular level corresponding to decrease of sphingosine and specific increase of C(14)- and C(16)-ceramide. LCL464 did not induce lysosomal destabilization nor degradation of ACDase, showed increased cell death demonstrating inherent anticancer activity in a wide range of different cancer cell lines, and induction of apoptosis via executioner caspases activation. LCL464 represents a novel structural lead as chemotherapeutic agent acting via the inhibition of ACDase. PMID- 19217790 TI - Medicinal chemistry approaches to the inhibition of dipeptidyl peptidase-4 for the treatment of type 2 diabetes. AB - Emerging as an epidemic of the 21st century type 2 diabetes has become a major health problem throughout the globe. The number of deaths attributable to diabetes reflects the insufficient glycemic control achieved with the treatments used in recent past. DPP-4 inhibitors have been investigated as a new therapy with novel mechanisms of action and improved tolerability. DPP-4, a protease that specifically cleaves dipeptides from proteins and oligopeptides after a penultimate N-terminal proline or alanine, is involved in the degradation of a number of neuropeptides, peptide hormones and cytokines, including the incretins GLP-1 and GIP. As soon as released from the gut in response to food intake, GLP-1 and GIP exert a potent glucose-dependent insulinotropic action, thereby playing a key role in the maintenance of post-meal glycemic control. Consequently, inhibiting DPP-4 prolongs the action of GLP-1 and GIP, which in turn improves glucose homeostasis with a low risk of hypoglycemia and potential for disease modification. Indeed, clinical trials involving diabetic patients have shown improved glucose control by administering DPP-4 inhibitors, thus demonstrating the benefit of this promising new class of antidiabetics. Intense research activities in this area have resulted in the launch of sitagliptin and vildagliptin (in Europe only) and the advancement of a few others into preregistration/phase 3, for example, saxagliptin, alogliptin and ABT-279. Achieving desired selectivity for DPP-4 over other related peptidases such as DPP 8 and DPP-9 (inhibition of which was linked to toxicity in animal studies) and long-acting potential for maximal efficacy (particularly in more severe diabetic patients) were the major challenges. Whether these goals are achieved with the present series of inhibitors in the advanced stages of clinical development is yet to be confirmed. Nevertheless, treatment of this metabolic disorder especially in the early stages of the disease via DPP-4 inhibition has been recognized as a validated principle and a large number of inhibitors are presently in various stage of pre-clinical/clinical development. Sitagliptin is a new weapon in the arsenal of oral antihyperglycemic agents. This review will focus on the journey of drug discovery of DPP-4 inhibitors for oral delivery covering a brief scientific background and medicinal chemistry approaches along with the status of advanced clinical candidates. PMID- 19217791 TI - Synthesis, cytotoxic activity, and DNA binding properties of antitumor cis-1,2 dihydroxy-1,2-dihydrobenzo[b]acronycine cinnamoyl esters. AB - Monocinnamoyl esters at position 2 of (+/-)-cis-1,2-dihydroxy-6-methoxy-3,3,14 trimethyl-1,2,3,14-tetrahydro-7H-benzo[b]pyrano[3,2-h]acridin-7-one and their acetyl derivatives at position 1 were prepared as stabilized analogues of the anticancer alkylating agent S23906-1. Monocinnamoyl esters at position 2 were slower DNA alkylators than the reference 2-monoacetate. Mixed esters bearing an acetyl ester group at position 1 and a cinnamoyl ester group at position 2 alkylated DNA slower than S23906-1. A strong correlation was observed between cytotoxicity and DNA alkylation kinetics, with slower alkylators displaying more potent antiproliferative activities. The most cytotoxic compounds proved to be significantly active in vivo against murine C-38 adenocarcinoma implanted in mice, but less potent than S23906-1. PMID- 19217792 TI - Dose profile modeling of Idaho National Laboratory's active neutron interrogation laboratory. AB - A new laboratory has been commissioned at Idaho National Laboratory for performing active neutron interrogation research and development. The facility is designed to provide radiation shielding for deuterium-tritium (DT) fusion (14.1 MeV) neutron generators (2 x 10(8) n/s), deuterium-deuterium (DD) fusion (2.5 MeV) neutron generators (1 x 10(7) n/s), and (252)Cf spontaneous fission neutron sources (6.96 x 10(7) n/s, 30 microg). Shielding at the laboratory is comprised of modular concrete shield blocks 0.76 m thick with tongue-in-groove features to prevent radiation streaming, arranged into one small and one large test vault. The larger vault is designed to allow operation of the DT generator and has walls 3.8m tall, an entrance maze, and a fully integrated electrical interlock system; the smaller test vault is designed for (252)Cf and DD neutron sources and has walls 1.9 m tall and a simple entrance maze. Both analytical calculations and numerical simulations were used in the design process for the building to assess the performance of the shielding walls and to ensure external dose rates are within required facility limits. Dose rate contour plots have been generated for the facility to visualize the effectiveness of the shield walls and entrance mazes and to illustrate the spatial profile of the radiation dose field above the facility and the effects of skyshine around the vaults. PMID- 19217793 TI - Identification and expression of a new delta-12 fatty acid desaturase (FAD2-4) gene in upland cotton and its functional expression in yeast and Arabidopsis thaliana plants. AB - A cotton (Gossypium hirsutum L.) genomic clone encompassing a 17.9-kb DNA fragment was found to contain a delta-12 fatty acid desaturase gene (designated FAD2-4). The FAD2-4 open reading frame has 1,155bp and is uninterrupted, encoding a conceptual FAD2-4 polypeptide of 384 amino acids that has 98% identity with the cotton FAD2-3 polypeptide. The FAD2-4 gene has a single intron of 2,780 bp in its 5'-untranslated region (5'-UTR). The 3'-flanking regions and 5'-UTR introns in the FAD2-4 and FAD2-3 genes are quite different, indicating that the genes might be paralogs in the cotton genome. Reverse transcriptase (RT)-PCR analysis indicated that the FAD2-4 and FAD2-3 genes were expressed in all tissues examined, including seeds, seedling tissues, young and mature leaves, roots, stems, developing flower buds, and ovule fibers. These constitutive patterns of expression were notably different from that of the FAD2-1 gene, which was restricted to seeds and developing flower buds, or to the expression of a newly identified FAD2-2 gene isoform, which was barely detectable in roots, hypocotyls, stems, and fibers, but was expressed in all other tissues. The FAD2-4 coding region was expressed in yeast and shown to encode a functional delta-12 desaturase, converting oleic acid (C18:1) to linoleic acid (C18:2) in recombinant yeast cells. In addition, both the FAD2-4 and the FAD2-3 genes were transferred into the Arabidopsis thaliana fad2-1 mutant background where they effectively restored wild type fatty acid composition and growth characteristics. Finally, the cotton FAD2-4 green fluorescent protein (GFP) fusion polypeptide appeared to be localized in the endomembrane system of transgenic Arabidopsis plants in the complemented fad2-1 mutant background, supporting a functional ER location for the cotton FAD2-4 polypeptide in this heterologous plant system. Thus, a new functional member of the FAD2 gene family in cotton has been characterized, indicating a complex regulation of membrane lipid desaturation in this important fiber/oilseed crop. PMID- 19217794 TI - Unilateral double epiphrenic diverticula of the esophagus. PMID- 19217796 TI - Splenic rupture as a presenting feature of endocarditis. AB - We describe the first case of infective endocarditis presenting with spontaneous splenic rupture. Our patient, a known intravenous drug user presented with hypovolaemic shock secondary to splenic rupture. The patient was resuscitated and underwent an emergency splenectomy. Subsequent clinical examination revealed a systolic murmur and a diagnosis of mitral valve infective endocarditis was made after echocardiography. Splenic tissue, blood cultures and mitral valve tissue all cultured Enterococcus faecalis. The patient had a successful mitral valve replacement and was discharged home after 44 days. To our knowledge this is the first reported case of enterococcal endocarditis presenting with splenic rupture. This case highlights the need to consider endocarditis in spontaneous splenic rupture particularly in those patients in a high risk group, such as IV drug users, especially if they lack a clear history of trauma. PMID- 19217795 TI - The effect of hypothermia on neuronal viability following cardiopulmonary bypass and circulatory arrest in newborn piglets. AB - OBJECTIVE: To determine the effect of recovery with mild hypothermia after cardiopulmonary bypass (CPB) and deep hypothermic circulatory arrest (DHCA) on the activity of selected key proteins involved in initiation (Bax, Caspase-3) or inhibition of apoptotic injury (Bcl-2, increased ratio Bcl-2/Bax) in the brain of newborn piglets. METHODS: The piglets were placed on CPB, cooled with pH-stat management to 18 degrees C, subjected to 30 min of DHCA followed by 1h of low flow at 20 ml/kg/min, rewarmed to 37 degrees C (normothermia) or to 33 degrees C (hypothermia), separated from CPB, and monitored for 6h. Expression of above proteins was measured in striatum, hippocampus and frontal cortex by Western blots. The results are mean for six experiments+/-SEM. RESULTS: There were no significant differences in Bcl-2 level between normothermic and hypothermic groups. The Bax levels in normothermic group in cortex, hippocampus and striatum were 94+/-9, 136+/-22 and 125+/-34 and decreased in the hypothermic group to 59+/ 17 (p=0.028), 70+/-6 (p=0.002) and 48+/-8 (p=0.01). In cortex, hippocampus and striatum Bcl-2/Bax ratio increased from 1.23, 0.79 and 0.88 in normothermia to 1.96, 1.28 and 2.92 in hypothermia. Expression of Caspase-3 was 245+/-39, 202+/ 74 and 244+/-31 in cortex, hippocampus and striatum in the normothermic group and this decreased to 146+/-24 (p=0.018), 44+/-16 (p=7 x 10(-7)) and 81+/-16 (p=0.01) in the hypothermic group. CONCLUSION: In neonatal piglet model of cardiopulmonary bypass with circulatory arrest, mild hypothermia during post bypass recovery provides significant protection from cellular apoptosis, as indicated by lower expression of Bax and Caspase-3 and an increased Bcl-2/Bax ratio. The biggest protection was observed in striatum probably by decreasing of neurotoxicity of striatal dopamine. PMID- 19217797 TI - The use of methylene blue for vasodilatory shock in a pediatric lung transplant patient. PMID- 19217798 TI - The feasibility of speckle tracking for intraoperative assessment of regional myocardial function by transesophageal echocardiography. AB - OBJECTIVES: The authors aimed to examine the feasibility of intraoperative transesophageal echocardiography (TEE) acquisition of a non-Doppler-based, speckle tracking-derived myocardial deformation parameter (strain) immediately before and after coronary artery bypass graft (CABG) surgery in patients with reduced left ventricular (LV) function. DESIGN: A clinical study. SETTING: The cardiac surgery operating room of a tertiary referral institution. PATIENTS: Ten patients with reduced LV function (ejection fraction lower than 35%) undergoing coronary revascularization were studied before and immediately after the procedure. INTERVENTIONS: Perioperative TEE. MEASUREMENTS AND RESULTS: A total of 120 myocardial segments were analyzed before and after CABG surgery. In visually obtained wall motion scoring (WMS), there were 29 normokinetic (N), 69 hypokinetic (H), 19 akinetic (A), and 3 dyskinetic (D) segments preoperatively and 26 N, 65 H, 21 A, and 8 D segments after CABG surgery. Preoperative radial strain correlated well with WMS (R = 0.82, p < 0.0001), whereas longitudinal strain showed only a weak correlation (R = 0.36, p < 0.0001). Postoperatively, correlations were similar. Interobserver variability as analyzed by kappa statistics showed better agreement for radial (kappa = 0.82 +/- 0.05, p = 0.001) and longitudinal strain (kappa = 0.73 +/- 0.06, p = 0.004) than for WMS (kappa = 0.65 +/- 0.06). Preoperatively, strain was markedly greater in normally perfused segments than in ischemic segments, whereas the mean WMS revealed only minor differences. CONCLUSIONS: Strain calculation from TEE images is feasible during cardiac surgery and correlates well with WMS but has better interobserver agreement. Strain analysis, but not WMS, detected wall motion differences between normally perfused and ischemic segments. This simple method allows objective intraoperative quantification of myocardial segment function and may become an important monitoring tool in the future. PMID- 19217799 TI - Left ventricular outflow tract to right atrial fistula after aortic valve replacement. PMID- 19217800 TI - A left ventricular-to-right atrial shunt in a patient with a perimembranous ventricular septal defect: role of intraoperative transesophageal echocardiography. PMID- 19217801 TI - Transesophageal echocardiography for verification of the position of the electrocardiographically-placed central venous catheter. AB - OBJECTIVE: Compare changes in P-wave amplitude of the intra-atrial electrocardiogram (ECG) and its corresponding transesophageal echocardiography (TEE)-controlled position to verify the exact localization of a central venous catheter (CVC) tip. DESIGN: A prospective study. SETTING: University, single institutional setting. PARTICIPANTS: Two hundred patients undergoing elective cardiac surgery. INTERVENTIONS: CVC placement via the right internal jugular vein with ECG control using the guidewire technique and TEE control in 4 different phases: phase 1: CVC placement with normalized P wave and measurement of distance from the crista terminalis to the CVC tip; phase 2: TEE-controlled placement of the CVC tip; parallel to the superior vena cava (SVC) and measurements of P-wave amplitude; phase 3: influence of head positioning on CVC migration; and phase 4: evaluation of positioning of the CVC postoperatively using a chest x-ray. MEASUREMENTS AND MAIN RESULTS: The CVC tip could only be visualized in 67 patients on TEE with a normalized P wave. In 198 patients with the CVC parallel to the SVC wall controlled by TEE (phase 2), an elevated P wave was observed. Different head movements led to no significant migration of the CVC (phase 3). On a postoperative chest-x-ray, the CVC position was correct in 87.6% (phase 4). CONCLUSION: The study suggests that the position of the CVC tip is located parallel to the SVC and 1.5 cm above the crista terminalis if the P wave starts to decrease during withdrawal of the catheter. The authors recommend that ECG control as per their study should be routinely used for placement of central venous catheters via the right internal jugular vein. PMID- 19217802 TI - Back pain and cardiogenic shock: an atypical diagnosis for typical symptoms. PMID- 19217803 TI - The usefulness of near-infrared spectroscopy for detecting and monitoring status epilepticus after pediatric cardiac surgery. PMID- 19217804 TI - The accuracy of the Vigileo/FloTrac system has been improved--follow-up after a software update: a blinded comparative study of 30 cardiosurgical patients. PMID- 19217805 TI - Global analysis of nuclear receptor expression and dysregulation in human osteoarthritic articular cartilage: reduced LXR signaling contributes to catabolic metabolism typical of osteoarthritis. AB - OBJECTIVE: Compare the expression and regulation of nuclear receptors (NRs) in osteoarthritic and normal human articular cartilage. METHOD: The transcriptional levels of 48 NRs and additional related proteins were measured in mRNA from human articular cartilage from subjects with osteoarthritis (OA) and compared to samples from subjects without OA, using microarrays, individual quantitative reverse transcriptase polymerase chain reaction assays, and a custom human NR TaqMan Low Density Array (TLDA). The functional effect of liver X receptor (LXR) activity in cartilage was studied by measuring proteoglycan (PG) synthesis and degradation in articular cartilage explant cultures following treatment with the synthetic LXR agonist T0901317. RESULTS: Thirty-one of 48 NRs analyzed by TLDA were found to be measurably expressed in human articular cartilage; 23 of these 31 NRs showed significantly altered expression in OA vs unaffected cartilage. Among these, LXRalpha and LXRbeta, and their heterodimeric partners retinoid X receptor (RXR)alpha and RXRbeta were all expressed at significantly lower levels in OA cartilage, as were LXR target genes ABCG1 and apolipoproteins D and E. Addition of LXR agonist to human OA articular chondrocytes and to cartilage explant cultures resulted in activation of LXR-mediated transcription and significant reduction of both basal and interleukin (IL)-1-mediated PG degradation. CONCLUSIONS: Articular cartilage expresses a substantial number of NRs, and a large proportion of the expressed NRs are dysregulated in OA. In particular, LXR signaling in OA articular cartilage is impaired, and stimulation of LXR transcriptional activity can counteract the catabolic effects of IL-1. We conclude that LXR agonism may be a possible therapeutic option for OA. PMID- 19217806 TI - Aquaretic inhibits renal cancer proliferation: Role of vasopressin receptor-2 (V2 R). AB - Vasopressin (AVP) is a hormone with antidiuretic properties that is also involved in cellular proliferation of breast, pulmonary, and pancreatic neoplasias, attributable to the interaction with specific receptors, among which is the V2-R. Using a culture model of CAKI-2 and A498 cancer cells, our study aimed to verify if renal carcinoma cells also express V2-R and whether receptor activation modulates their proliferation. Immunofluorescence and RT-PCR showed that both CAKI-2 and A498 cells effectively synthesize and express the V2-R. Administration of the vasopressin analogue DDAVP induced an evident growth in both CAKI-2 and A498 cell lines. However, this proliferative effect was completely avoided by the preventive addition of the V2-R antagonist SR121463B (satavaptan). Our study shows for the first time that renal cancer may effectively synthesize and express the V2-R. Furthermore, AVP exerts in vitro a proliferative effect by acting on this receptor, as the selective V2-R blockage is able to completely prevent the cellular growth. A validation of these findings with in vivo models is required to ascertain if the eventual presence of V2-R could influence the aggressiveness of human renal neoplasias. From this point of view, a new, interesting therapeutical application of V2-R antagonists in the treatment of renal cancer could also be proposed, similar to that successfully described in the treatment of autosomal polycystic kidney disease (ADPKD). PMID- 19217807 TI - Postconditioning: current controversies and clinical implications. AB - OBJECTIVES: Postconditioning of ischaemic tissue, via mechanical or pharmacological manipulation, offers an exciting avenue towards amelioration of ischaemia-reperfusion injury. Born from the concept of ischaemic preconditioning, postconditioning is advantageous in that prior knowledge of the ischaemic insult is not required, and thus clinical translation may be further reaching. This review explores the current evidence and controversies in both animal and human studies and multiple organ systems. METHODS: A Medline search was conducted to identify English-language articles with 'postconditioning' as a keyword. Two independent researchers scrutinised the literature search for potentially relevant articles. Reference lists from selected articles were manually searched for further relevant articles. RESULTS AND CONCLUSIONS: Postconditioning has been shown to be successful in reducing ischaemia-reperfusion injury in both animal models and clinical trials. Human studies are presently limited to cardiac studies, but there is scope for research into other organ systems with potential beneficial effects, particularly within the field of vascular surgery where ischaemia-reperfusion occurs by nature of both - the disease and the intervention. PMID- 19217808 TI - Letter to the Editor re: indication for intervention in chronic type B aortic dissection. PMID- 19217809 TI - Identification of a novel deletion in UDP-glucuronosyltransferase gene in a patient with Crigler-Najjar syndrome type I. PMID- 19217810 TI - Use of a stand-alone pressure plate for the objective evaluation of forelimb symmetry in sound ponies at walk and trot. AB - Subtle lameness in horses may be difficult to diagnose and methods to evaluate lameness objectively are useful when equine clinicians fail to reach a consensus. The aim of this study was to determine whether equine pressure plate measurements are repeatable when used to calculate forelimb loading (peak vertical pressure [PVP], peak vertical force [PVF], vertical impulse [VI]) and symmetry ratios, and to establish if these data are similar to the 'gold standard' force plate values. Since plate dimensions are relatively small, ponies were used to enable recordings to be taken from both forelimbs in one trial. Five sound ponies were walked and trotted over a pressure plate which was embedded in a custom-made runway. For each pony, five valid trials were recorded during two different days. A trial was considered valid if complete prints of both fore hooves were recorded consecutively and if velocity was within a preset range. The PVP, PVF and VI values showed an acceptable variability (CV < or = 16%), with PVF (130% of bodyweight [BW], n=5) similar to previously reported force plate data (128% BW, n=48). Mean symmetry ratios appeared to be high (> 95%) and showed a low variability (CV < 5%). The stand-alone pressure plate permitted adequate registration of forelimb PVF, VI and limb symmetry with a high level of precision in sound ponies. PMID- 19217811 TI - Effects of L-spin longitudinal quadrupolar relaxation in S[L] heteronuclear recoupling and S-spin magic-angle spinning NMR. AB - In experiments on S-L heteronuclear spin systems with evolution of the S-spin magnetization under the influence of a quadrupolar nucleus (L-spin), effects of longitudinal quadrupolar (T(1Q)) relaxation of the L-spin coherence on the sub millisecond time scale have been documented and explored, and methods for minimizing their effect have been demonstrated. The longitudinal relaxation results in heteronuclear dephasing even in the reference signal S(0) of S[L] REDOR, REAPDOR, RIDER, or SPIDER experiments, due to T(1Q)-relaxation of the transiently generated S(y)L(z) coherence, reducing or even eliminating the observable dephasing DeltaS. Pulse sequences for measuring an improved reference signal S(00) with minimal heteronuclear recoupling but the same number of pulses as for S(0) and S have been demonstrated. From the observed intensity DeltaS(0)=S(00)-S(0) and the SPIDER signal DeltaS/S(0), T(1Q) can be estimated. Accelerated decays analogous to the dipolar S(0) curves will occur in T(2) measurements for J-coupled S-L spin pairs. Even in the absence of recoupling pulses, fast T(1Q) relaxation of the unobserved nucleus shortens the transverse relaxation time T(2S,MAS) of the observed nucleus, in particular at low spinning frequencies, due to unavoidable heteronuclear dipolar evolution during a rotation period. The observed spinning-frequency dependence of T(2S,MAS) matches the theoretical prediction and may be used to estimate T(1Q). The effects are demonstrated on several (13)C[(14)N] spin systems, including an arginine derivative, the natural N-acetylated polysaccharide chitin, and a model peptide, (POG)(10). PMID- 19217812 TI - IGF-IR/ERK content and response to IGF-I and insulin in adipocytes from small for gestational age children. AB - BACKGROUND/AIMS: The GH-IGF-I axis plays an important role on pre/postnatal growth in humans, and IGF-I regulates cell proliferation, differentiation, and metabolic homeostasis. The adipose tissue has an important function in energy storage, and plays an endocrine role through the production of several hormones and growth factors. There are few studies in humans regarding IGF-I and Insulin receptor signaling in adipocytes, particularly from AGA and SGA children. METHODS: We studied 22 healthy prepubertal children (6.1+/-0.4 years), born at term (11 SGA and 11 AGA), and normal BMI at the time of the study. Primary cell cultures were established from subcutaneous adipose tissue biopsies. Preadipocytes were differentiated and stimulated with IGF-I or insulin and we studied IGF-IR, IR, AKT, and ERK content and phosphorylation. RESULTS: The SGA children were shorter than the AGA children (height SDS -2.14+/-0.11 vs. 0.02+/ 0.19, p<0.05). A lower content of IGF-IR, IR, AKT, and ERK was observed in adipocytes from SGA compared with AGA children. IGF-I stimulation increased IGF IR and ERK phosphorylation in adipocytes from AGA, but not from SGA children. CONCLUSION: The lower content and reduced phosphorylation of IGF-I signaling observed in adipocytes from SGA children may be related to the metabolic abnormalities described in these children. PMID- 19217813 TI - A point mutation in the neu1 promoter recruits an ectopic repressor, Nkx3.2 and results in a mouse model of sialidase deficiency. AB - SM/J is an inbred mouse strain with a complex phenotype including small body size, impaired immune response and a tissue-specific sialidase deficiency. We identified a regulatory mutation, (-519G-->A) within the neu1 promoter which in reporter assays resulted in significantly reduced transcription. This mutation generates a consensus binding site for Nkx3 family transcription repressors. Recombinant Nkx3.2 bound strongly to and preferentially repressed transcription of the mutant promoter. This tissue-specific deficiency results in a retarded immune response and modulates leukocyte recruitment. Examination of the hepatic microcirculation in mutant mice revealed increased rolling and decreased adhesion of leukocytes. Our findings support a significant role for lysosomal sialidase in inflammation and highlight the significance of repressor-recruitment in genetic disease. PMID- 19217814 TI - The paradox of the carnitine palmitoyltransferase type Ia P479L variant in Canadian Aboriginal populations. AB - Investigation of seven patients from three families suspected of a fatty acid oxidation defect showed mean CPT-I enzyme activity of 5.9+/-4.9 percent of normal controls. The families, two Inuit, one First Nation, live in areas of Canada geographically very distant from each other. The CPT1 and CPT2 genes were fully sequenced in 5 of the patients. All were homozygous for the same P479L mutation in a highly conserved region of the CPT1 gene. Two patients from the first family were also homozygous for the CPT2 F352C polymorphism in the CPT2 gene. Genotyping the patients and their family members confirmed that all seven patients were homozygous for the P479L variant allele in the CPT1 gene, as were 27 of 32 family members. Three of the seven patients and two cousins had hypoketotic hypoglycemia attributable to CPT-Ia deficiency, but adults homozygous for the variant denied hypoglycemia. We screened 422 consecutive newborns from the region of one of the Inuit families for this variant; 294 were homozygous, 103 heterozygous, and only 25 homozygous normal; thus the frequency of this variant allele is 0.81. There was an infant death in one family and at least 10 more deaths in those infants (7 homozygous, 3 heterozygous) consecutively tested for the mutation at birth. Thus there is an astonishingly high frequency of CPT1 P479L variant and, judging from the enzyme analysis in the seven patients, also CPT-I deficiency in the areas of Canada inhabited by these families. Despite the deficiency of CPT-Ia which is the major rate-limiting enzyme for long chain fatty acid oxidation, clinical effects, with few exceptions, were slight or absent. One clue to explaining this paradox is that, judging from the fatty acid oxidation studies in whole blood and fibroblasts, the low residual activity of CPT-Ia is sufficient to allow a reasonable flux through the mitochondrial oxidation system. It is likely that the P479L variant is of ancient origin and presumably its preservation must have conveyed some advantage. PMID- 19217815 TI - Participation of asparagine 370 and glutamine 235 in the catalysis by acid beta glucosidase: the enzyme deficient in Gaucher disease. AB - The hydrolysis of glucosylceramide by acid beta-glucosidase proceeds via a two step, double displacement mechanism that includes cleavage of the O-beta glucosidic bond, enzyme-glucosylation and, then, enzyme-deglucosylation. Two residues that may impact this cycle are N370 and E235. The N370S mutant enzyme is very common in Gaucher disease type 1 patients. Homology and crystal data predictions suggested that E235 is the acid/base catalyst in the hydrolytic reaction. Here, the roles of N370 and E235 in hydrolysis were explored using mutant proteins with selected amino acid substitutions. Heterologously expressed enzymes were characterized using inhibitors, activators, and alternative substrates to gain insight into the effects on the glucosylation (single turnover) and deglucosylation (transglucosylation) steps in catalysis. Specific substitutions at N370 selectively altered only the glucosylation step whereas N370S altered this and the deglucosylation steps. To provide functional data to support E235 as the acid/base catalyst, progress curves with poor substrates with more acidic leaving groups were used in the presence and absence of azide as an exogenous nucleophile. The restoration of E235G activity to nearly wild-type levels was achieved using azide with 2,4-dinitrophenyl-beta-glucoside as substrate. The loss of the acidic arm of the pH optimum activity curve of E235G provided additional functional support for E235 as the acid/base in catalysis. This study provides insight into the function of these residues in acid beta glucosidase active site function. PMID- 19217816 TI - Bone tissue and hyperhomocysteinemia. AB - Bone tissue quality is determined not only by multiple architectural variables, but also by the mechanical properties of collagen type 1. Homocysteinuria is a genetic disease whose manifestations include severe hyperhomocysteinemia and decreased bone strength. The effects of smaller homocysteine elevations on bone tissue are difficult to demonstrate in clinical studies. Studies in animals and in humans suggest that homocysteine may weaken collagen crosslinks and, if present in large amounts, interfere with bone remodeling. Whether routine homocysteine assays should be performed to detect bone frailty remains unclear. In clinical practice, the focus should be on identifying patients with potential causes of homocysteine elevation (e.g., medications), who should then be given vitamin D and folic acid supplementation if needed. This approach may improve not only bone health, but also vascular and general health. PMID- 19217817 TI - Effect of resistance exercise on bone mineral density in premenopausal women. AB - OBJECTIVE: To assess the effect of 9 months of strength training on total body and regional bone mineral density (BMD, g/cm(2)) in 58 premenopausal women aged 30-50 years. METHODS: Participants were randomized to either twice weekly supervised strength training for 15 weeks followed by 24 weeks of unsupervised training (treatment group) or control group. Height, weight, maximal muscular strength, nutrient intake and physical activity were assessed. Total body dual energy X-ray absorptiometry (DXA, Lunar Prodigy) scans were taken and analyzed for body composition (lean and fat mass), and BMD for total body and its sub regions (spine, hip, arms and legs). All measurements were performed at baseline, 15 and 39 weeks. Analysis of covariance was used to assess group differences in BMD change adjusted for baseline BMD, weight, energy and calcium intake. RESULTS: At baseline, the two groups had similar BMD and body size characteristics ( P<0.05 for all), except that the treatment group had lower body weight (-7.1 kg), and higher energy (+259 kJ/d) and calcium (+232 mg/d) intake at baseline. Adjusted % change in BMD over 15 weeks (0.5% vs. 0.4%) or 39 weeks (0.9% vs. 1.2%) did not differ significantly between the exercise and control groups, respectively. The exercise group increased BMD at the spine and legs (1-2.2%), while there was no change in the controls, but differences between groups were not significant. CONCLUSION: Strength training over 9 months did not lead to significantly greater change in total body or regional BMD in premenopausal women. PMID- 19217818 TI - Regional variation in orthopedic surgery in Switzerland. AB - The study systematically describes the frequency and geographic variability of major surgical interventions for musculoskeletal disorders in Switzerland. Age- and sex-standardized rates for joint replacements, arthroscopies, spine surgery and hip fracture repair were calculated for hospital service regions. Various statistical analyses were used to measure the extent of variation. The authors argue that the surgery of hip fractures can be used as index surgery in the context of analyzing variations in orthopedic surgery. Temporal trends imply that patient demand and supply factors related to clinical ambiguity and non-medical incentives of providers are far more important components leading to increased use than the sole effect of an aging population. PMID- 19217819 TI - Place, positionality, and priorities: experts' views on women's health at the Mexico-US border. AB - Improving health care in the Mexican-US transborder region presents challenges not only of harmonization of systems but differences in values and expectations for policies. We explore the contrasting and overlapping views of health experts in both countries regarding the ways in which geographic location, scale, and professional roles shape their notions of the region, priorities for women's health, and interpretations of the socio-cultural concept of gender in relation to health. The study is based on interviews with legislators, health administrators, health advocates, and researchers on each side of the border. We suggest ways in which more nuanced perspectives might be brought to bear in the policy discourses on the region. PMID- 19217820 TI - From the family universe to the outside world: family relations, school attitude, and perception of racism in Caribbean and Filipino adolescents. AB - Caribbean and Filipino immigrant families in Canada have much in common: the women have often immigrated as domestic workers, first-generation children may be separated from their parents for long periods, and they must deal with negative stereotypes of their ethnic group. This transcultural study looks at the associations between family relations and adolescents' perceptions of both their own group and the host society, and analyzes how these affect their mental health. The results suggest that family cohesion plays a key role in shaping adolescents' perceptions of racism in the host country and in promoting a positive appraisal of their own community, thus highlighting the need for a systemic understanding of family and intergroup relations. PMID- 19217821 TI - Spatio-temporal clustering of cholera: the impact of flood control in Matlab, Bangladesh, 1983-2003. AB - Introducing flood control to an area of endemic waterborne diseases could have significant impacts on spatio-temporal occurrence of cholera. Using 21-year data from Bangladesh, we conducted cluster analysis to explore changes in spatial and temporal distribution of cholera incidence since the construction of flood control structures. Striking changes in temporal cluster patterns emerged, including a shift from dry-season to rainy-season clusters following flood protection and delayed clustering inside the protected areas. Spatial differences in pre-flood protection and post-protection cholera clusters are weaker. Changes in spatio-temporal cholera clustering, associated with implementation of flood protection strategies, could affect local cholera prevention efforts. PMID- 19217822 TI - The eye contact effect: mechanisms and development. AB - The 'eye contact effect' is the phenomenon that perceived eye contact with another human face modulates certain aspects of the concurrent and/or immediately following cognitive processing. In addition, functional imaging studies in adults have revealed that eye contact can modulate activity in structures in the social brain network, and developmental studies show evidence for preferential orienting towards, and processing of, faces with direct gaze from early in life. We review different theories of the eye contact effect and advance a 'fast-track modulator' model. Specifically, we hypothesize that perceived eye contact is initially detected by a subcortical route, which then modulates the activation of the social brain as it processes the accompanying detailed sensory information. PMID- 19217823 TI - Association between DNA double strand break gene Ku80 polymorphisms and oral cancer susceptibility. AB - The DNA double strand break repair gene Ku80 is thought to play a major role in the caretaking of the overall genome stability. It is very possible that defective in double strand break repair capacity can lead to human carcinogenesis. Thus, the polymorphic variants of Ku80 were firstly investigated regarding their association with oral cancer susceptibility. In this hospital based case-control study, the association of Ku80 promoter G-1401T (rs828907), promoter C-319T (rs11685387), and intron19 (rs9288518) polymorphisms with oral cancer risk in a Taiwanese population was investigated. 600 patients with oral cancer and 600 age- and gender-matched healthy controls recruited were genotyped and analyzed by PCR-RFLP method. There were significant differences between oral cancer and control groups in the distributions of their genotypes (P=0.0038) and allelic frequencies (P=0.0044) in the Ku80 promoter G-1401T polymorphism. In the other two polymorphisms, there was no difference between both groups in the distribution of either genotype or allelic frequency. There is a synergistic gene environmental interaction between Ku80 and areca chewing. Compared with G/G genotype in Ku80 promoter G-1401T, the G/T plus T/T significantly enhanced the risk only in the areca chewers (odds ratio=1.603; 95% confidence interval=1.053 2.011), not in the non-areca chewers. In conclusion, the Ku80 promoter G-1401T is correlated with oral cancer susceptibility and this polymorphism may be a useful marker for oral cancer prevention and early detection. PMID- 19217825 TI - Granule-associated serine proteases: granzymes might not just be killer proteases. AB - The cytotoxic cell granule secretory pathway is viewed as indispensable for eliminating tumor and virally infected cells through a process in which the pore forming protein, perforin, delivers the serine protease granzymes into cells targeted for destruction. Residing in cytotoxic cells, granzymes were originally anticipated to act both extracellularly and intracellularly. With the discovery that isolated granzymes induce apoptosis when combined with perforin, the broader functionality of the granzymes became unattractive. The purpose of this article is to describe observations indicating that granzymes possess non-cytotoxic activities that might include such diverse biologic effects as stimulation of pro inflammatory cytokines, remodeling of extracellular matrices and inactivation of intracellular pathogens. PMID- 19217824 TI - AMPed up immunity: how antimicrobial peptides have multiple roles in immune defense. AB - Antimicrobial peptides (AMPs) are widely expressed and rapidly induced at epithelial surfaces to repel assault from diverse infectious agents including bacteria, viruses, fungi and parasites. Much information suggests that AMPs act by mechanisms that extend beyond their capacity to serve as gene-encoded antibiotics. For example, some AMPs alter the properties of the mammalian membrane or interact with its receptors to influence diverse cellular processes including cytokine release, chemotaxis, antigen presentation, angiogenesis and wound healing. These functions complement their antimicrobial action and favor resolution of infection and repair of damaged epithelia. Opposing this, some microbes have evolved mechanisms to inactivate or avoid AMPs and subsequently become pathogens. Thus, AMPs are multifunctional molecules that have a central role in infection and inflammation. PMID- 19217826 TI - Using differential gene expression to study Entamoeba histolytica pathogenesis. AB - The release of the Entamoeba histolytica genome has facilitated the development of techniques to survey rapidly and to relate gene expression with biology. The association and potential contribution of differential gene expression to the life cycle and the virulence of this protozoan parasite of humans are reviewed here. PMID- 19217827 TI - DevR-mediated adaptive response in Mycobacterium tuberculosis H37Ra: links to asparagine metabolism. AB - The DevR transcriptional switch that defines the response of Mycobacterium tuberculosis to the lack of oxygen is now well established and likely helps the bacteria shift to a state of persistence. The M. tuberculosis two component signal transduction system (TCS), DevR-DevS, implicated in this transition to latency, is differentially expressed in H37Ra and H37Rv strains. Despite originating from the H37 ancestral strain, H37Ra and H37Rv have significant differences in their growth, physiology, and virulence. To further dissect the role of DevR in growth adaptive processes of M. tuberculosis, we investigated the hypoxic response of the avirulent H37Ra strain. Our results show that the DevR DevS TCS in H37Ra is responsive to hypoxia and capable of target gene regulation, indicating similar DevR-DevS signaling pathways in H37Ra and H37Rv. A key finding of this study was the constitutive expression of the Rv3134c-devR-devS operon and a subset of sentinel DevR-regulated genes in aerobic cultures of H37Ra but not H37Rv grown in Dubos-Tween-albumin medium. Asparagine and/or catabolites of asparagine metabolism were implicated in aerobic induction of the DevR-DevS TCS in H37Ra. This is the first report of medium-specific constitutive expression of the DevR regulon in an avirulent strain and suggests a potential role for metabolite(s) in the activation of the DevR-DevS TCS. PMID- 19217830 TI - Special section on autoimmune autonomic ganglionopathy. Editorial. PMID- 19217831 TI - Relationship between sympathetic nervous activity and inflammatory response after subarachnoid hemorrhage in a perforating canine model. AB - The objective of the present study was to evaluate the correlation between sympathetic nerve activation and inflammatory response in the acute stage of subarachnoid hemorrhage (SAH) in a canine perforating model. SAH was induced by perforation of the basilar artery with the use of a microcatheter via the femoral artery in 20 mongrel dogs. Hemodynamic parameters and intracranial pressure were recorded, and blood sample for C3a, C5b-9, IL-6, IL-8 and noradrenaline kinetic determination were measured at 0, 5, 15, 30, 60, 120, and 180 min after SAH. Noradrenaline (pg/mL) increased abruptly from 104+/-59 to 2010+/-918 at 5 min after SAH. C3a and C5b-9 reached peak values at 15 min and IL-6 and IL-8 reached peak values at 30 min after SAH, respectively. The peak values of C3a and C5b-9 correlated positively with the peak value of noradrenaline (r=0.743 and r=0.753, respectively). The peak values of IL-6 and IL-8 also correlated positively with the peak values of noradrenaline (r=0.603 and r=0.681, respectively).These results suggest that a pronounced activation of the sympathetic nervous system and the inflammatory response occurs in acute stage of SAH. Significant association between the rate of spillover of norepinephrine to plasma and the plasma levels of inflammatory markers indicates that the two processes, sympathetic activation and immune response are quantitatively linked in early stage after SAH. The exact mechanisms underlying this phenomenon deserved further investigations. PMID- 19217832 TI - The Yin and Yang of the MMS21-SMC5/6 SUMO ligase complex in homologous recombination. AB - Maintaining genomic stability is critical for the prevention of disease. Numerous DNA repair pathways help to maintain genomic stability by correcting potentially lethal or disease-causing lesions to our genomes. Mounting evidence suggests that the post-translational modification sumoylation plays an important regulatory role in several aspects of DNA repair. The E3 SUMO ligase MMS21/NSE2 has gained increasing attention for its function in homologous recombination (HR), an error free DNA repair pathway that mediates repair of double-strand breaks (DSBs) using the sister chromatid as a repair template. MMS21/NSE2 is part of the SMC5/6 complex, which has been shown to facilitate DSB repair, collapsed replication fork restart, and telomere elongation by HR. Here, I review the function of the SMC5/6 complex and its associated MMS21/NSE2 SUMO ligase activity in homologous recombination. PMID- 19217833 TI - Regulating post-translational modifications of the eukaryotic replication clamp PCNA. AB - Modifications of the eukaryotic sliding clamp, proliferating cell nuclear antigen (PCNA), by ubiquitin and the ubiquitin-related protein SUMO, are well known to influence the choice of pathways for the processing of DNA lesions during replication. Over the past few years, significant progress has been made not only with respect to the molecular consequences that each of the modifications has for the properties of PCNA, but also in terms of the cellular signals that elicit the ubiquitylation or sumoylation of PCNA in the appropriate situations. This review will discuss the regulatory mechanisms that control PCNA modifications, emphasizing the important role of the DNA template on which PCNA acts in activating the relevant ubiquitin and SUMO conjugation factors, and pointing out similarities as well as some interesting variations among different organisms in the regulation of PCNA modifications. PMID- 19217834 TI - A shortcut from plasma to chromatographic analysis: straightforward and fast sample preparation for analysis of green tea catechins in human plasma. AB - This paper describes a new and straightforward method for determination of the green tea catechins epicatechin, epicatechin gallate, epigallocatechin, and epigallocatechin gallate in human plasma. Sample preparation includes addition only of dimethylformamide and trichloroacetic acid. After centrifugation, the supernatant can be injected into the HPLC. If required, the glucuronides and sulphates of the catechins can be enzymatically hydrolysed before extraction. Recovery ranges from 92.9 to 98.2%; limits of detection, from 2.4 to 5.0 ng/mL; and relative standard deviations, from 3.1 to 8.6%. Twelve samples can be processed within 45 min, and are then ready to be injected into the HPLC. The method was successfully applied to human plasma. This method is suitable for studies on absorption, bioavailability, and kinetics of green tea catechins in plasma. Since manual work and time consumption are minimal, the procedure is especially useful for large numbers of samples. PMID- 19217835 TI - Automated metal-free multiple-column nanoLC for improved phosphopeptide analysis sensitivity and throughput. AB - We report on the development and characterization of automated metal-free multiple-column nanoLC instrumentation for sensitive and high-throughput analysis of phosphopeptides with mass spectrometry. The system employs a multiple-column capillary LC fluidic design developed for high-throughput analysis of peptides (Anal. Chem. 2001, 73, 3011-3021), incorporating modifications to achieve broad and sensitive analysis of phosphopeptides. The integrated nanoLC columns (50 microm i.d. x 30 cm containing 5 microm C18 particles) and the on-line solid phase extraction columns (150 microm i.d. x 4 cm containing 5 microm C18 particles) were connected to automatic switching valves with non-metal chromatographic accessories, and other modifications to avoid the exposure of the analyte to any metal surfaces during handling, separation, and electrospray ionization. The nanoLC developed provided a separation peak capacity of approximately 250 for phosphopeptides (and approximately 400 for normal peptides). A detection limit of 0.4 fmol was obtained when a linear ion trap tandem mass spectrometer (Finnegan LTQ) was coupled to a 50-microm i.d. column of the nanoLC. The separation power and sensitivity provided by the nanoLC-LTQ enabled identification of approximately 4600 phosphopeptide candidates from approximately 60 microg COS-7 cell tryptic digest followed by IMAC enrichment and approximately 520 tyrosine phosphopeptides from approximately 2mg of human T cells digests followed by phosphotyrosine peptide immunoprecipitation. PMID- 19217836 TI - The evolution of oesophageal function testing and its clinical applications in the management of patients with oesophageal disorders. AB - The last decade has brought significant technical advances in laparoscopic surgery. In this constantly evolving technological climate, less told is the story of the evolution of diagnostic technology that improved the clinical management of patients with oesophageal disorders. The successful outcome of the laparoscopic treatment of oesophageal disorders is due to a combination of three different factors: the skills and the ability of the foregut surgeon, the high volume of referral, the expertise in the critical evaluation of the oesophageal function tests. This is an art per se, and it is rarely acknowledged in the achievement of the excellent results of surgery. Oesophageal function testing provides the clinician with information that cannot be obtained by the clinical, endoscopic, and radiological evaluation of patients. This expertise, intimately coupled with the other factors, allows the surgeon to better understand the pathophysiology of these diseases and to provide the optimal management. Therefore, it is essential to understand the evolution that this technology is currently undergoing, and how these changes are expanding the current indications for antireflux surgery by identifying additional predictors of successful outcome. PMID- 19217837 TI - Endoclipping in spurting diverticular haemorrhage. PMID- 19217838 TI - Cellular response of preosteoblasts to nanograined/ultrafine-grained structures. AB - Metallic materials with submicron- to nanometer-sized grains provide surfaces that are different from conventional polycrystalline materials because of the large proportion of grain boundaries with high free energy. In the study described here, the combination of cellular and molecular biology, materials science and engineering advances our understanding of cell-substrate interactions, especially the cellular activity between preosteoblasts and nanostructured metallic surfaces. Experiments on the effect of nano-/ultrafine grains have shown that cell attachment, proliferation, viability, morphology and spread are favorably modulated and significantly different from conventional coarse-grained structures. Additionally, immunofluorescence studies demonstrated stronger vinculin signals associated with actin stress fibers in the outer regions of the cells and cellular extensions on nanograined/ultrafine-grained substrate. These observations suggest enhanced cell-substrate interaction and activity. The differences in the cellular response on nanograined/ultrafine grained and coarse-grained substrates are attributed to grain size and degree of hydrophilicity. The outcomes of the study are expected to reduce challenges to engineer bulk nanostructured materials with specific physical and surface properties for medical devices with improved cellular attachment and response. The data lay the foundation for a new branch of nanostructured materials for biomedical applications. PMID- 19217839 TI - Epigenetic effects of glucocorticoids. AB - The early nurturing environment has persistent influences on developmental programming of inter-individual differences in metabolic and endocrine function that contribute to emotional and cognitive performance through life. These effects are mediated, in part, through neonatal programming of hypothalamic pituitary-adrenal (HPA) axis function. Animal models support this hypothesis. For example, in the rat natural variations in maternal care influence HPA axis stress reactivity in the offspring via long-term changes in tissue-specific gene expression. Studies in vivo and in vitro show that maternal licking and grooming increases glucocorticoid receptor expression in the offspring via increased hippocampal serotonergic tone accompanied by increased histone acetylase transferase activity, histone acetylation and DNA demethylation mediated by the transcription factor nerve growth factor-inducible protein-A. These effects are reversed by early postnatal cross-fostering and by pharmacological manipulations, including trichostatin A (TSA) and l-methionine administration in adulthood. These studies demonstrate that an epigenetic state of a gene can be established through early in life experience, and is potentially reversible in adult life. Accordingly, epigenetic modifications in target gene promoters in response to environmental demand may ensure stable yet dynamic regulation that mediates persistent changes in biological and behavioral phenotype over the lifespan. PMID- 19217840 TI - Peripheral arterial- bypass grafts in the hand or foot in systemic sclerosis. AB - Although the literature suggests that sympathectomy alone may improve ischaemic digital pain, fingertip ulceration and cold intolerance in vaso-occlusive patients, questions regarding long-term efficacy remain. Two adult patients, aged 42 and 49 years, had severe digital ischaemia and ulceration in a finger and a toe, respectively, and arteriograms confirmed the occlusions of the superficial palmar arch and tarsal arch, respectively. Microsurgical re-vascularisation of the hand and foot and arch reconstruction were performed by using the greater saphenous vein for reconstruction. Following successful vascular reconstruction, digital temperatures, pain and cold sensitivity and health-related quality of life improved significantly. No recurrence has been encountered after X and Y months of follow-up. PMID- 19217841 TI - [Adult Schonlein-Henoch purpura associated with epidermoid carcinoma of the lung]. AB - INTRODUCTION: Association between Schonlein-Henoch purpura and neoplasm can suggest the responsibility of tumour antigens in the genesis of the vasculitis. We report a new case of squamous cell carcinoma associated with Schonlein-Henoch purpura and we discuss the reality of this association. CASE REPORT: We report the case of a 50-year-old man who presents Schonlein-Henoch purpura with a purpura of lower limbs, joint involvement, gastrointestinal lesions and IgA renal mesangial deposits. The patient received three intravenous methylprednisolone pulses followed with oral corticosteroids. Six months later, while the vasculitis was in remission, the patient presented a squamous cell lung carcinoma. He was treated by chemotherapy and local radiotherapy. At the late follow-up, the neoplasm was incompletely resolved. CONCLUSION: The neoplasm could be responsible of the development of the Schonlein-Henoch purpura. The discovery of this systemic vasculitis in an elderly patient should warrant a deep screening for an occult neoplasm. PMID- 19217842 TI - Energy landscape for DNA rotation and sliding through a phage portal. AB - Molecular motors involved in the packaging of DNA in tailed viruses are among the strongest known. The mechanism by which the motors operate has long been speculated to involve a coupling between rotation of the portal pore (the gate through which DNA passes upon its packaging or ejection), and translation of DNA. Recent experimental evidence rules out portal rotation with a substantial degree of certainty. We have created an atomistic model for the interaction between DNA and the portal of the bacteriophage SPP1, on the basis of cryo-electron microscopy images and of a recently solved crystal structure. A free energy surface describing the interaction is calculated using molecular dynamics simulations, and found to be inconsistent with a mechanism in which portal rotation drives DNA import. The low-energy pathways on the surface are used to advance a hypothesis on DNA import compatible with all available experiments. Additionally, temperature-dependent kinetic data are used to validate computed barriers to DNA ejection. PMID- 19217843 TI - Nanoelectromechanics of methylated DNA in a synthetic nanopore. AB - Methylation of cytosine is a covalent modification of DNA that can be used to silence genes, orchestrating a myriad of biological processes including cancer. We have discovered that a synthetic nanopore in a membrane comparable in thickness to a protein binding site can be used to detect methylation. We observe a voltage threshold for permeation of methylated DNA through a <2 nm diameter pore, which we attribute to the stretching transition; this can differ by >1 V/20 nm depending on the methylation level, but not the DNA sequence. PMID- 19217844 TI - Effects of multiple occupancy and interparticle interactions on selective transport through narrow channels: theory versus experiment. AB - Many biological and artificial transport channels function without direct input of metabolic energy during a transport event and without structural rearrangements involving transitions from a closed to an open state. Nevertheless, such channels are able to maintain efficient and selective transport. It has been proposed that attractive interactions between the transported molecules and the channel can increase the transport efficiency and that the selectivity of such channels can be based on the strength of the interaction of the specifically transported molecules with the channel. Herein, we study the transport through narrow channels in a framework of a general kinetic theory, which naturally incorporates multiparticle occupancy of the channel and non-single-file transport. We study how the transport efficiency and the probability of translocation through the channel are affected by interparticle interactions in the confined space inside the channel, and establish conditions for selective transport. We compare the predictions of the model with the available experimental data and find good semiquantitative agreement. Finally, we discuss applications of the theory to the design of artificial nanomolecular sieves. PMID- 19217845 TI - Dynamics and energetics: a consensus analysis of the impact of calcium on EF-CaM protein complex. AB - We have studied the relationship between dynamical correlations and energetic contributions in an attempt to model the transmission of information inside protein-protein complexes. The complex formed between the edema factor (EF) of Bacillus anthracis and calmodulin (CaM) was taken as an example, as the formation and stability of the complex depend on the calcium complexation level. The effect of calcium through EF-CaM residue network has been investigated with various approaches: 1), the elastic network model; 2), the local feature analysis; 3), the generalized correlations; and 4), the energetic dependency maps (EDMs), on 15 ns molecular dynamics simulations of the complex loaded with 0, 2, or 4 Ca2+ ions. The elastic network model correctly describes the basic architecture of the complex but is poorly sensitive to the level of calcium compared to the other methods. The local feature analysis allows us to characterize the local dynamics of the complex and the propagation of the calcium signal through CaM. The analyses of global dynamics and energetics--through generalized correlations and EDMs--provide a comprehensive picture of EF-CaM architecture and can be unified by using the concept of residue network connectedness. A medium connectedness, defined as the ability of each residue to communicate with all remaining parts of the complex, is observed for the 2Ca2+ level, which was experimentally identified as the most stable form of EF-CaM. The hierarchy of relative stabilities given by the EDMs sheds a new light on the EF-CaM interaction mechanism described experimentally and supports an organization of the complex architecture centered around nucleation points. PMID- 19217846 TI - Parameter sensitivity analysis in electrophysiological models using multivariable regression. AB - Computational models of electrical activity and calcium signaling in cardiac myocytes are important tools for understanding physiology. The sensitivity of these models to changes in parameters is often not well-understood, however, because parameter evaluation can be a time-consuming, tedious process. I demonstrate here what I believe is a novel method for rapidly determining how changes in parameters affect outputs. In three models of the ventricular action potential, parameters were randomized, repeated simulations were run, important outputs were calculated, and multivariable regression was performed on the collected results. Random parameters included both maximal rates of ion transport and gating variable characteristics. The procedure generated simplified, empirical models that predicted outputs resulting from new sets of input parameters. The linear regression models were quite accurate, despite nonlinearities in the mechanistic models. Moreover, the regression coefficients, which represent parameter sensitivities, were robust, even when parameters were varied over a wide range. Most importantly, a side-by-side comparison of two similar models identified fundamental differences in model behavior, and revealed model predictions that were both consistent with, and inconsistent with, experimental data. This new method therefore shows promise as a tool for the characterization and assessment of computational models. The general strategy may also suggest methods for integrating traditional quantitative models with large scale data sets obtained using high-throughput technologies. PMID- 19217847 TI - Biochemistry on a leash: the roles of tether length and geometry in signal integration proteins. AB - We use statistical mechanics and simple ideas from polymer physics to develop a quantitative model of proteins whose activity is controlled by flexibly tethered ligands and receptors. We predict how the properties of tethers influence the function of these proteins and demonstrate how their tether length dependence can be exploited to construct proteins whose integration of multiple signals can be tuned. One case study to which we apply these ideas is that of the Wiskott Aldrich Syndrome Proteins as activators of actin polymerization. More generally, tethered ligands competing with those free in solution are common phenomena in biology, making this an important specific example of a widespread biological idea. PMID- 19217848 TI - Ions and inhibitors in the binding site of HIV protease: comparison of Monte Carlo simulations and the linearized Poisson-Boltzmann theory. AB - Proteins can be influenced strongly by the electrolyte in which they are dissolved, and we wish to model, understand, and ultimately control such ionic effects. Relatively detailed Monte Carlo (MC) ion simulations are needed to capture biologically important properties of ion channels, but a simpler treatment of ions, the linearized Poisson-Boltzmann (LPB) theory, is often used to model processes such as binding and folding, even in settings where the LPB theory is expected to be inaccurate. This study uses MC simulations to assess the reliability of the LPB theory for such a system, the constrained, anionic active site of HIV protease. We study the distributions of ions in and around the active site, as well as the energetics of displacing ions when a protease inhibitor is inserted into the active site. The LPB theory substantially underestimates the density of counterions in the active site when divalent cations are present. It also underestimates the energy cost of displacing these counterions, but the error is not consequential because the energy cost is less than kBT, according to the MC calculations. Thus, the LPB approach will often be suitable for studying energetics, but the more detailed MC approach is critical when ionic distributions and fluxes are at issue. PMID- 19217850 TI - A biophysically based mathematical model for the kinetics of mitochondrial calcium uniporter. AB - Ca2+ transport through mitochondrial Ca2+ uniporter is the primary Ca2+ uptake mechanism in respiring mitochondria. Thus, the uniporter plays a key role in regulating mitochondrial Ca2+. Despite the importance of mitochondrial Ca2+ to metabolic regulation and mitochondrial function, and to cell physiology and pathophysiology, the structure and composition of the uniporter functional unit and kinetic mechanisms associated with Ca2+ transport into mitochondria are still not well understood. In this study, based on available experimental data on the kinetics of Ca2+ transport via the uniporter, a mechanistic kinetic model of the uniporter is introduced. The model is thermodynamically balanced and satisfactorily describes a large number of independent data sets in the literature on initial or pseudo-steady-state influx rates of Ca2+ via the uniporter measured under a wide range of experimental conditions. The model is derived assuming a multi-state catalytic binding and Eyring's free-energy barrier theory-based transformation mechanisms associated with the carrier-mediated facilitated transport and electrodiffusion. The model is a great improvement over the previous theoretical models of mitochondrial Ca2+ uniporter in the literature in that it is thermodynamically balanced and matches a large number of independently published data sets on mitochondrial Ca2+ uptake. This theoretical model will be critical in developing mechanistic, integrated models of mitochondrial Ca2+ handling and bioenergetics which can be helpful in understanding the mechanisms by which Ca2+ plays a role in mediating signaling pathways and modulating mitochondrial energy metabolism. PMID- 19217849 TI - Correlated motions and interactions at the onset of the DNA-induced partial unfolding of Ets-1. AB - The binding of the Ets-1 transcription factor to its target DNA sequence is characterized by a highly unusual conformational change consisting of the unfolding of inhibitory helix 1 (HI-1). To probe the interactions that lead to this unfolding, we performed molecular dynamics simulations of the folded states of apo-Ets-1 and the Ets-1-DNA complex. The simulations showed large differences in correlated motions between helix 4 (H4) and HI-1. In apo-Ets-1, H4 and HI-1 moved in-phase and stabilized each other by hydrogen bonding and macrodipolar interactions, whereas in the DNA-bound state, the motion was out-of-phase, with a disruption of the stabilizing interactions. This change in motion was due to hydrogen-bonding interactions between helix 1 (H1) and the DNA. The dipolar energy between H1 and H4 was modulated by hydrogen bonds between H1 and DNA, and, in accordance with experiments, elimination of the hydrogen bonds increased the stability of HI-1. The simulations confirm that the hydrogen bonds between H1 and DNA act as a conformational switch and show that the presence of DNA is communicated from H1 to H4, destabilizing HI-1. The calculations reveal a critical role for correlated motions at the onset of the DNA-induced unfolding. PMID- 19217851 TI - Effect of macromolecular crowding on reaction rates: a computational and theoretical study. AB - The effect of macromolecular crowding on the rates of association reactions are investigated using theory and computer simulations. Reactants and crowding agents are both hard spheres, and when two reactants collide they form product with a reaction probability, p(rxn). A value of p(rxn) < 1 crudely mimics the fact that proteins must be oriented properly for an association reaction to occur. The simulations show that the dependence of the reaction rate on the volume fraction of crowding agents varies with the reaction probability. For reaction probabilities close to unity where most of encounters between reactants lead to a reaction, the reaction rate always decreases as the volume fraction of crowding agents is increased due to the reduced diffusion coefficient of reactants. On the other hand, for very small reaction probabilities where, in most of encounters, the reaction does not occur, the reaction rate increases with the volume fraction of crowding agents--in this case, due to the increase probability of a recollision. The Smoluchowski theory refined with the radiation boundary condition and the radial distribution function at contact is in quantitative agreement with simulations for the reaction rate constant and allows the quantitative analysis of both effects separately. PMID- 19217852 TI - Theoretical analysis of disruptions in DNA minicircles. AB - Under sufficient bending stress, which appears in DNA minicircles and small DNA loops, the double helix experiences local disruptions of its regular structure. We developed a statistical-mechanical treatment of the disruptions in DNA minicircles, studied experimentally by Du et al. The model of disruptions used in our Monte Carlo simulation of minicircle conformations specifies these conformations by three parameters: DNA bend angle at the disruption, theta(d); local DNA unwinding caused by the disruption; and the free energy associated with the disruption in the unstressed double helix, G(d). The model is applicable to any structural type of disruption, kinks or opening of single basepairs. The simulation shows that accounting for both torsional and bending deformation associated with the disruptions is very important for proper analysis. We obtained a relationship between values of G(d) and theta(d) under which the simulation results are compatible with the experimental data. The relationship suggests that the free energy of basepair opening, which includes flipping out both bases, is significantly higher than the generally accepted value. The model is also applied to the analysis of j-factors of very short DNA fragments. PMID- 19217853 TI - Mechanical properties of the icosahedral shell of southern bean mosaic virus: a molecular dynamics study. AB - The mechanical properties of viral shells are crucial for viral assembly and infection. To study their distribution and heterogeneity on the viral surface, we performed atomistic force-probe molecular dynamics simulations of the complete shell of southern bean mosaic virus, a prototypical T = 3 virus, in explicit solvent. The simulation system comprised more than 4,500,000 atoms. To facilitate direct comparison with atomic-force microscopy (AFM) measurements, a Lennard Jones sphere was used as a model of the AFM tip, and was pushed with different velocities toward the capsid protein at 19 different positions on the viral surface. A detailed picture of the spatial distribution of elastic constants and yielding forces was obtained that can explain corresponding heterogeneities observed in previous AFM experiments. Our simulations reveal three different deformation regimes: a prelinear regime of outer surface atom rearrangements, a linear regime of elastic capsid deformation, and a rearrangement regime that describes irreversible structural changes and the transition from elastic to plastic deformation. For both yielding forces and elastic constants, a logarithmic velocity dependency is evident over nearly two decades, the explanation for which requires including nonequilibrium effects within the established theory of enforced barrier crossing. PMID- 19217854 TI - Low energy defibrillation in human cardiac tissue: a simulation study. AB - We aim to assess the effectiveness of feedback-controlled resonant drift pacing as a method for low energy defibrillation. Antitachycardia pacing is the only low energy defibrillation approach to have gained clinical significance, but it is still suboptimal. Low energy defibrillation would avoid adverse side effects associated with high voltage shocks and allow the application of implantable cardioverter defibrillator (ICD) therapy, in cases where such therapy is not tolerated today. We present results of computer simulations of a bidomain model of cardiac tissue with human atrial ionic kinetics. Reentry was initiated and low energy shocks were applied with the same period as the reentry, using feedback to maintain resonance. We demonstrate that such stimulation can move the core of reentrant patterns, in the direction that depends on the location of the electrodes and the time delay in the feedback. Termination of reentry is achieved with shock strength one-order-of-magnitude weaker than in conventional single shock defibrillation. We conclude that resonant drift pacing can terminate reentry at a fraction of the shock strength currently used for defibrillation and can potentially work where antitachycardia pacing fails, due to the feedback mechanisms. Success depends on a number of details that these numerical simulations have uncovered. PMID- 19217855 TI - Changes in negative charge at the luminal mouth of the pore alter ion handling and gating in the cardiac ryanodine-receptor. AB - We have tested the hypothesis that a high density of negative charge at the luminal mouth of the RyR2 pore plays a pivotal role in the high cation conductance and limited selectivity observed in this channel by introducing into each monomer a double point mutation to neutralize acidic residues in this region of the mouse RyR2 channel. The resultant channel, ED4832AA, is capable of functioning as a calcium-release channel in situ. Consistent with our hypothesis, the ED4832AA mutation altered the ion handling characteristics of single RyR2 channels. The mutant channel retains the ability to discriminate between cations and anions but cation conductance is altered significantly. Unitary K+ conductance is reduced at low levels of activity but increases dramatically as activity is raised and shows little sign of saturation. ED4832AA no longer discriminates between divalent and monovalent cations. In addition, the gating characteristics of single RyR2 channels are altered markedly by residue neutralization. Open probability in the ED4832AA channel is substantially higher than that of the wild-type channel. Moreover, at holding potentials in excess of +/-50 mV several subconductance states become apparent in ED4832AA and are more prevalent at very high holding potentials. These observations are discussed within the structural framework provided by a previously developed model of the RyR2 pore. Our data indicates that neutralization of acidic residues in the luminal mouth of the pore produces wide-ranging changes in the electric field in the pore, the interaction energies of permeant ions in the pore and the stability of the selectivity filter region of the pore, which together contribute to the observed changes ion handling and gating. PMID- 19217856 TI - Doxorubicin inactivates myocardial cytochrome c oxidase in rats: cardioprotection by Mito-Q. AB - Doxorubicin (DOX) is used for treating various cancers. Its clinical use is, however, limited by its dose-limiting cardiomyopathy. The exact mechanism of DOX induced cardiomyopathy still remains unknown. The goals were to investigate the molecular mechanism of DOX-induced cardiomyopathy and cardioprotection by mitoquinone (Mito-Q), a triphenylphosphonium-conjugated analog of coenzyme Q, using a rat model. Rats were treated with DOX, Mito-Q, and DOX plus Mito-Q for 12 weeks. The left ventricular function as measured by two-dimensional echocardiography decreased in DOX-treated rats but was preserved during Mito-Q plus DOX treatment. Using low-temperature ex vivo electron paramagnetic resonance (EPR), a time-dependent decrease in heme signal was detected in heart tissues isolated from rats administered with a cumulative dose of DOX. DOX attenuated the EPR signals characteristic of the exchange interaction between cytochrome c oxidase (CcO)-Fe(III) heme a3 and CuB. DOX and Mito-Q together restored these EPR signals and the CcO activity in heart tissues. DOX strongly downregulated the stable expression of the CcO subunits II and Va and had a slight inhibitory effect on CcO subunit I gene expression. Mito-Q restored CcO subunit II and Va expressions in DOX-treated rats. These results suggest a novel cardioprotection mechanism by Mito-Q during DOX-induced cardiomyopathy involving CcO. PMID- 19217857 TI - Peptide adsorption to lipid bilayers: slow processes revealed by linear dichroism spectroscopy. AB - The adsorption and insertion kinetics for the association of two 34-residue cyclic peptides with phosphocholine membranes have been studied using circular and linear dichroism approaches. The two peptides studied are identical with the exception of two residues, which are both tyrosine in one of the peptides and tryptophan in the other. Both peptides adopt random coil conformations in solution in the absence of membranes and do not aggregate at concentrations below 20 microM. After addition to liposome dispersions, circular dichroism spectroscopy indicated that both peptides undergo an extremely rapid transformation to a beta-conformation that remains unchanged throughout the remainder of the experiment. Linear dichroism (LD) spectroscopy was used to study the kinetics of membrane adsorption and insertion. The data were analyzed by nonlinear least squares approaches, leading to identification of a number of bound states and their corresponding LD spectra. Two pseudo-first order processes could be identified that were common to both peptides. The first occurred with a time constant of the order of 1 min and led to a bound state characterized by weak LD signals, with significant bands corresponding to the transitions of aromatic side chains. The second process occurred with an unusually long time constant of between 75 and 100 min, forming a state with considerably stronger positive LD absorbance in the far-ultraviolet region of the spectrum. For the tyrosine-substituted peptide, a third slow process with a long time constant (76 min) could also be delineated and was attributed to rearrangements of the peptide within the membrane. PMID- 19217858 TI - Membrane protein frustration: protein incorporation into hydrophobic mismatched binary lipid mixtures. AB - Bacteriophage M13 major coat protein was reconstituted in different nonmatching binary lipid mixtures composed of 14:1PC and 22:1PC lipid bilayers. Challenged by this lose-lose situation of hydrophobic mismatch, the protein-lipid interactions are monitored by CD and site-directed spin-label electron spin resonance spectroscopy of spin-labeled site-specific single cysteine mutants located in the C-terminal protein domain embedded in the hydrophobic core of the membrane (I39C) and at the lipid-water interface (T46C). The CD spectra indicate an overall alpha helical conformation irrespective of the composition of the binary lipid mixture. Spin-labeled protein mutant I39C senses the phase transition in 22:1PC, in contrast to spin-labeled protein mutant T46C, which is not affected by the transition. The results of both CD and electron spin resonance spectroscopy clearly indicate that the protein preferentially partitions into the shorter 14:1PC both above and below the gel-to-liquid crystalline phase transition temperature of 22:1PC. This preference is related to the protein tilt angle and energy penalty the protein has to pay in the thicker 22:1PC. Given the fact that in Escherichia coli, which is the host for M13 bacteriophage, it is easier to find shorter 14 carbon acyl chains than longer 22 carbon acyl chains, the choice the M13 coat protein makes seems to be evolutionary justified. PMID- 19217859 TI - Pulmonary surfactant model systems catch the specific interaction of an amphiphilic peptide with anionic phospholipid. AB - Interfacial behavior was studied in pulmonary surfactant model systems containing an amphiphilic alpha-helical peptide (Hel 13-5), which consists of 13 hydrophobic and five hydrophilic amino acid residues. Fully saturated phospholipids of dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylglycerol (DPPG) were utilized to understand specific interactions between anionic DPPG and cationic Hel 13-5 for pulmonary functions. Surface pressure (pi)-molecular area (A) and surface potential (DeltaV)-A isotherms of DPPG/Hel 13-5 and DPPC/DPPG (4:1, mol/mol)/Hel 13-5 preparations were measured to obtain basic information on the phase behavior under compression and expansion processes. The interaction leads to a variation in squeeze-out surface pressures against a mole fraction of Hel 13-5, where Hel 13-5 is eliminated from the surface on compression. The phase behavior was visualized by means of Brewster angle microscopy, fluorescence microscopy, and atomic force microscopy. At low surface pressures, the formation of differently ordered domains in size and shape is induced by electrostatic interactions. The domains independently grow upon compression to high surface pressures, especially in the DPPG/Hel 13-5 system. Under the further compression process, protrusion masses are formed in AFM images in the vicinity of squeeze out pressures. The protrusion masses, which are attributed to the squeezed-out Hel 13-5, grow larger in lateral size with increasing DPPG content in phospholipid compositions. During subsequent expansion up to 35 mN m(-1), the protrusions retain their height and lateral diameter for the DPPG/Hel 13-5 system, whereas the protrusions become smaller for the DPPC/Hel 13-5 and DPPC/DPPG/Hel 13-5 systems due to a reentrance of the ejected Hel 13-5 into the surface. In this work we detected for the first time, to our knowledge, a remarkably large hysteresis loop for cyclic DeltaV-A isotherms of the binary DPPG/Hel 13-5 preparation. This exciting phenomenon suggests that the specific interaction triggers two completely independent processes for Hel 13-5 during repeated compression and expansion: 1), squeezing-out into the subsolution; and 2), and close packing as a monolayer with DPPG at the interface. These characteristic processes are also strongly supported by atomic force microscopy observations. The data presented here provide complementary information on the mechanism and importance of the specific interaction between the phosphatidylglycerol headgroup and the polarized moiety of native surfactant protein B for biophysical functions of pulmonary surfactants. PMID- 19217861 TI - Primary conformation change in bacteriorhodopsin on photoexcitation. AB - Ultrafast dynamics of bacteriorhodopsin (bR) has been extensively studied experimentally and theoretically. However, there are several contradictory results reported, indicating that its detailed dynamics and initial process have not yet been fully clarified. In this work, changes in the amplitude and phase of molecular vibration in the isomerization process of bR were real-time probed simultaneously at 128 different wavelengths through intensity modulation of the electronic transition. Systematic information on the transient change in continuous spectrum extending from 505 nm (2.45 eV) to 675 nm (1.84 eV) showed different dynamics in each spectral region reflecting the difference in the excited states and intermediates dominating the dynamics during the photoisomerization. Careful analysis of the transient spectral changes and spectrograms calculated from the vibrational real-time traces elucidated that the primary event just after photoexcitation is the deformation of the retinal configuration, which decays within 30 fs near the C=N bond in the protonated Schiff base. The intensity of C=N stretching mode starts to decrease before the initiation of the frequency modulation of the C=C stretching mode. The C=C stretching mode frequency was modulated by a coupled torsion around the C13=C14 bond, leading to the photoisomerization around the bond. This study clarified the dynamics of the C=N and C=C stretching modes working as key vibration modes in the photoisomerization of bR. Furthermore, we have elucidated the modulation and decay dynamics of the C=C stretching mode in the photoreaction starting from H (Franck-Condon excited state) followed by I (twisted excited), and J (first intermediate) states. PMID- 19217860 TI - Force-generating cross-bridges during ramp-shaped releases: evidence for a new structural state. AB - Mechanical and two-dimensional (2D) x-ray diffraction studies suggest that during isometric steady-state contraction, strongly bound cross-bridges mostly occupy early states in the power stroke, whereas rigor or rigor-like cross-bridges could not be detected. However, it remained unclear whether cross-bridges accumulate, at least transiently, in rigor or rigor-like states in response to rapid-length releases. We addressed this question using time-resolved recording of 2D x-ray diffraction patterns of permeabilized fibers from rabbit psoas muscles during isometric contraction and when small, ramp-shaped length-releases were applied to these fibers. This maneuver allows a transient accumulation of cross-bridges in states near the end of their power stroke. By lowering the temperature to 5 degrees C, force transients were slowed sufficiently to record diffraction patterns in several 2-4-ms time frames before and during such releases, using the RAPID detector (Refined ADC Per Input Detector) at beam line ID02 of the European Synchrotron Radiation Facility (Grenoble, France). The same sequence of frames was recorded in relaxation and rigor. Comparisons of 2D patterns recorded during isometric contraction, with patterns recorded at different [MgATPgammaS] and at 1 degrees C, showed that changes in intensity profiles along the first and sixth actin layer lines (ALL1 and ALL6, respectively) allowed for discernment of the formation of rigor or rigor-like cross-bridges. During ramp-shaped releases of activated fibers, intensity profiles along ALL1 and ALL6 did not reveal evidence for the accumulation of rigor-like cross-bridges. Instead, changes in the ALL6 profile suggest that during ramp-shaped releases, cross-bridges transiently accumulate in a structural state that, to our knowledge, was not previously seen, but that could well be a strongly bound state with the light-chain binding domain in a conformation between a near prepower-stroke (isometric) orientation and the orientation in rigor. PMID- 19217862 TI - Time-resolved Fourier transform infrared study on photoadduct formation and secondary structural changes within the phototropin LOV domain. AB - Phototropins are plant blue-light photoreceptors containing two light-, oxygen-, or voltage-sensitive (LOV) domains and a C-terminal kinase domain. The two LOV domains bind noncovalently flavin mononucleotide as a chromophore. We investigated the photocycle of fast-recovery mutant LOV2-I403V from Arabidopsis phototropin 2 by step-scan Fourier transform infrared spectroscopy. The reaction of the triplet excited state of flavin with cysteine takes place with a time constant of 3 micros to yield the covalent adduct. Our data provide evidence that the flavin is unprotonated in the productive triplet state, disfavoring an ionic mechanism of bond formation. An intermediate adduct species was evident that displayed changes in secondary structure in the helix or loop region, and relaxed with a time constant of 120 micros. In milliseconds, the final adduct state is formed by further alterations of secondary structure, including beta-sheets. A comparison with wild-type adduct spectra shows that the mutation does not interfere with the functionality of the domain. All signals originate from within the LOV domain, because the construct does not comprise the adjacent Jalpha helix required for signal transduction. The contribution of early and late adduct intermediates to signal transfer to the Jalpha helix outside of the domain is discussed. PMID- 19217863 TI - The photocycle and proton translocation pathway in a cyanobacterial ion-pumping rhodopsin. AB - The genome of thylakoidless cyanobacterium Gloeobacter violaceus encodes a fast cycling rhodopsin capable of light-driven proton transport. We characterize the dark state, the photocycle, and the proton translocation pathway of GR spectroscopically. The dark state of GR contains predominantly all-trans-retinal and, similar to proteorhodopsin, does not show the light/dark adaptation. We found an unusually strong coupling between the conformation of the retinal and the site of Glu132, the homolog of Asp96 of BR. Although the photocycle of GR is similar to that of proteorhodopsin in general, it differs in accumulating two intermediates typical for BR, the L-like and the N-like states. The latter state has a deprotonated cytoplasmic proton donor and is spectrally distinct from the strongly red-shifted N intermediate known for proteorhodopsin. The proton uptake precedes the release and occurs during the transition to the O intermediate. The proton translocation pathway of GR is similar to those of other proton-pumping rhodopsins, involving homologs of BR Schiff base proton acceptor and donor Asp85 and Asp96 (Asp121 and Glu132). We assigned a pair of FTIR bands (positive at 1749 cm(-1) and negative at 1734 cm(-1)) to the protonation and deprotonation, respectively, of these carboxylic acids. PMID- 19217864 TI - Buffer interference with protein dynamics: a case study on human liver fatty acid binding protein. AB - Selection of suitable buffer types is often a crucial step for generating appropriate protein samples for NMR and x-ray crystallographic studies. Although the possible interaction between MES buffer (2-(N-morpholino)ethanesulfonic acid) and proteins has been discussed previously, the interaction is usually thought to have no significant effects on the structures of proteins. In this study, we demonstrate the direct, albeit weak, interaction between MES and human liver fatty acid binding protein (hLFABP). Rather than affecting the structure of hLFABP, we found that the dynamics of hLFABP, which were previously proposed to be relevant to its functions, were significantly affected by the binding of hLFABP with MES. Buffer interference with protein dynamics was also demonstrated with Bis-Tris buffer, which is quite different from MES and fatty acids in terms of their molecular structures and properties. This result, to our knowledge, is the first published report on buffer interference with protein dynamics on a microsecond to millisecond timescale and could represent a generic problem in the studies of functionally relevant protein dynamics. Although being a fortuity, our finding of buffer-induced changes in protein dynamics offers a clue to how hLFABP accommodates its ligands. PMID- 19217865 TI - Direct correlation between molecular dynamics and enzymatic stability: a comparative neutron scattering study of native human butyrylcholinesterase and its "aged" soman conjugate. AB - An incoherent elastic neutron scattering study of the molecular dynamics of native human butyrylcholinesterase and its "aged" soman-inhibited conjugate revealed a significant change in molecular flexibility on an angstrom-nanosecond scale as a function of temperature. The results were related to the stability of each state as established previously by differential scanning calorimetry. A striking relationship was found between the denaturation behavior and the molecular flexibility of the native and inhibited enzymes as a function of temperature. This was reflected in a quantitative correlation between the atomic mean-square displacements on an angstrom-nanosecond scale determined by neutron spectroscopy and the calorimetric specific heat. By the application of a simple two-state model that describes the transition from a folded to a denatured state, the denaturation temperatures of the native and the inhibited enzyme were correctly extracted from the atomic mean-square displacements. Furthermore, the transition entropy and enthalpy extracted from the model fit of the neutron data were, within the experimental accuracy, compatible with the values determined by differential scanning calorimetry. PMID- 19217866 TI - Thermodynamic effects of noncoded and coded methionine substitutions in calmodulin. AB - The methionine residues in the calcium (Ca2+) regulatory protein calmodulin (CaM) are structurally and functionally important. They are buried within the N- and C domains of apo-CaM but become solvent-exposed in Ca2+-CaM, where they interact with numerous target proteins. Previous structural studies have shown that methionine substitutions to the noncoded amino acids selenomethionine, ethionine, or norleucine, or mutation to leucine do not impact the main chain structure of CaM. Here we used differential scanning calorimetry to show that these substitutions enhance the stability of both domains, with the largest increase in melting temperature (19-26 degrees C) achieved with leucine or norleucine in the apo-C-domain. Nuclear magnetic resonance spectroscopy experiments also revealed the loss of a slow conformational exchange process in the Leu-substituted apo-C domain. In addition, isothermal titration calorimetry experiments revealed considerable changes in the enthalpy and entropy of target binding to apo-CaM and Ca2+-CaM, but the free energy of binding was largely unaffected due to enthalpy entropy compensation. Collectively, these results demonstrate that noncoded and coded methionine substitutions can be accommodated in CaM because of the structural plasticity of the protein. However, adjustments in side-chain packing and dynamics lead to significant differences in protein stability and the thermodynamics of target binding. PMID- 19217867 TI - Tightening the knot in phytochrome by single-molecule atomic force microscopy. AB - A growing number of proteins have been shown to adopt knotted folds. Yet the biological roles and biophysical properties of these knots remain poorly understood. We used protein engineering and atomic force microscopy to explore the single-molecule mechanics of the figure-eight knot in the chromophore-binding domain of the red/far-red photoreceptor, phytochrome. Under load, apo phytochrome unfolds at forces of approximately 47 pN, whereas phytochrome carrying its covalently bound tetrapyrrole chromophore unfolds at approximately 73 pN. These forces are not unusual in mechanical protein unfolding, and thus the presence of the knot does not automatically indicate a superstable protein. Our experiments reveal a stable intermediate along the mechanical unfolding pathway, reflecting the sequential unfolding of two distinct subdomains in phytochrome, potentially the GAF and PAS domains. For the first time (to the best of our knowledge), our experiments allow a direct determination of knot size under load. In the unfolded chain, the tightened knot is reduced to 17 amino acids, resulting in apparent shortening of the polypeptide chain by 6.2 nm. Steered molecular-dynamics simulations corroborate this number. Finally, we find that covalent phytochrome dimers created for these experiments retain characteristic photoreversibility, unexpectedly arguing against a dramatic rearrangement of the native GAF dimer interface upon photoconversion. PMID- 19217868 TI - Kinetics of contact formation and end-to-end distance distributions of swollen disordered peptides. AB - Unstructured polypeptide chains are subject to various degrees of swelling or compaction depending on the combination of solvent condition and amino acid sequence. Highly denatured proteins generally behave like random-coils with excluded volume repulsion, whereas in aqueous buffer more compact conformations have been observed for the low-populated unfolded state of globular proteins as well as for naturally disordered sequences. To quantitatively account for the different mechanisms inducing the swelling of polypeptides, we have examined three 14-residues peptides in aqueous buffer and in denaturant solutions, including the well characterized AGQ repeat as a reference and two variants, in which we have successively introduced charged side chains and removed the glycines. Quenching of the triplet state of tryptophan by close contact with cysteine has been used in conjunction with Forster resonance energy transfer to study the equilibrium and kinetic properties of the peptide chains. The experiments enable accessing end-to-end root mean-square distance, probability of end-to-end contact formation and intrachain diffusion coefficient. The data can be coherently interpreted on the basis of a simple chain model with backbone angles obtained from a library of coil segments of proteins and hard sphere repulsion at each Calpha position. In buffered water, we find that introducing charges in a glycine-rich sequence induces a mild chain swelling and a significant speed-up of the intrachain dynamics, whereas the removal of the glycines results in almost a two-fold increase of the chain volume and a drastic slowing down. In denaturants we observe a pronounced swelling of all the chains, with significant differences between the effect of urea and guanidinium chloride. PMID- 19217869 TI - Branching in amyloid fibril growth. AB - Using the peptide hormone glucagon and Abeta(1-40) as model systems, we have sought to elucidate the mechanisms by which fibrils grow and multiply. We here present real-time observations of growing fibrils at a single-fibril level. Growing from preformed seeds, glucagon fibrils were able to generate new fibril ends by continuously branching into new fibrils. To our knowledge, this is the first time amyloid fibril branching has been observed in real-time. Glucagon fibrils formed by branching always grew in the forward direction of the parent fibril with a preferred angle of 35-40 degrees . Furthermore, branching never occurred at the tip of the parent fibril. In contrast, in a previous study by some of us, Abeta(1-40) fibrils grew exclusively by elongation of preformed seeds. Fibrillation kinetics in bulk solution were characterized by light scattering. A growth process with branching, or other processes that generate new ends from existing fibrils, should theoretically give rise to different fibrillation kinetics than growth without such a process. We show that the effect of adding seeds should be particularly different in the two cases. Our light scattering data on glucagon and Abeta(1-40) confirm this theoretical prediction, demonstrating the central role of fibril-dependent nucleation in amyloid fibril growth. PMID- 19217870 TI - Oleoylethanolamide-based lyotropic liquid crystals as vehicles for delivery of amino acids in aqueous environment. AB - We have investigated the phase behavior of self-assembled lyotropic liquid crystals (LC) formed by ternary mixtures of oleoylethanolamide (OEA), water and arginine. OEA, a natural analog of the endogenous cannabinoid anandamide involved in the peripheral regulation of feeding, was selected as a main component due to its capacity to induce efficient decreases in food intake and gains in body mass. Arginine was selected as representative hydrophilic amino acid and added to the OEA-water mixture at different concentrations. The phase diagrams were determined by combining cross-polarized optical microscopy and small angle x-ray scattering. First, the phase diagram for the OEA-water system was determined. It was shown that these two compounds give rise to reverse Ia3d double gyroid and reverse Pn3m double diamond cubic phases existing in bulk over a large window of temperature and composition, and that for water content beyond 25% Pn3m coexisted with excess water. Successively, the influence of arginine as guest molecule in the water channels of the reverse LC was investigated. For the sake of comparison, results for the OEA-water-arginine system were compared with analog series of OEA-water glucose. The results showed that, at a fixed water content and temperature, the phase behavior of the liquid crystalline phases is strongly dependent on arginine concentration. In more detail, arginine could be encapsulated in the bulk OEA water LC up to 2.0% wt, whereas transitions from Ia3d to Pn3m cubic phase were observed with increasing arginine concentration. Interestingly, upon an increase of water concentration beyond 20-25%, Pn3m phase started to coexist with excess water releasing the arginine in external water solution. Quantitative measurements of arginine content inside the LC water channels and in the excess external water solution revealed a complete release of the amino acid, demonstrating that the investigated lyotropic liquid crystalline systems can be used as ideal vehicles for the delivery of functional hydrophilic active molecules in aqueous environment. PMID- 19217871 TI - Structure of functional Staphylococcus aureus alpha-hemolysin channels in tethered bilayer lipid membranes. AB - We demonstrate a method for simultaneous structure and function determination of integral membrane proteins. Electrical impedance spectroscopy shows that Staphylococcus aureus alpha-hemolysin channels in membranes tethered to gold have the same properties as those formed in free-standing bilayer lipid membranes. Neutron reflectometry provides high-resolution structural information on the interaction between the channel and the disordered membrane, validating predictions based on the channel's x-ray crystal structure. The robust nature of the membrane enabled the precise localization of the protein within 1.1 A. The channel's extramembranous cap domain affects the lipid headgroup region and the alkyl chains in the outer membrane leaflet and significantly dehydrates the headgroups. The results suggest that this technique could be used to elucidate molecular details of the association of other proteins with membranes and may provide structural information on domain organization and stimuli-responsive reorganization for transmembrane proteins in membrane mimics. PMID- 19217872 TI - DNA as membrane-bound ligand-receptor pairs: duplex stability is tuned by intermembrane forces. AB - We use membrane-anchored DNA as model adhesion receptors between lipid vesicles. By studying the thermal stability of DNA duplex formation, which tethers the vesicles into superstructures, we show that the melting temperature of a 10-base DNA sequence is dependent on the lipid composition of the tethered vesicles. We propose a simple model that describes how the intermembrane interactions tilt the free energy landscape for DNA binding. From our model, we estimate the area per DNA in the binding sites between vesicles and also the total area of the adhesion plaques. We find that vesicles containing a small proportion of cationic lipid that are modified with membrane-anchored DNA can be reversibly tethered by specific DNA interactions and that the DNA also induces a small attraction between these membranes, which stabilizes the DNA duplex. By increasing the equilibrium intermembrane distance on binding, we show that intermembrane interactions become negligible for the binding thermodynamics of the DNA and hence the thermal stability of vesicle aggregates becomes independent of lipid composition at large enough intervesicle separations. We discuss the implications of our findings with regards to cell adhesion and fusion receptors, and the programmable self-assembly of nano-structured materials by DNA hybridization. PMID- 19217873 TI - Rheology and confocal reflectance microscopy as probes of mechanical properties and structure during collagen and collagen/hyaluronan self-assembly. AB - In this work, the gelation of three-dimensional collagen and collagen/hyaluronan (HA) composites is studied by time sweep rheology and time lapse confocal reflectance microscopy (CRM). To investigate the complementary nature of these techniques, first collagen gel formation is investigated at concentrations of 0.5, 1.0, and 1.5 mg/mL at 37 degrees C and 32 degrees C. The following parameters are used to describe the self-assembly process in all gels: the crossover time (t(c)), the slope of the growth phase (k(g)), and the arrest time (t(a)). The first two measures are determined by rheology, and the third by CRM. A frequency-independent rheological measure of gelation, t(g), is also measured at 37 degrees C. However, this quantity cannot be straightforwardly determined for gels formed at 32 degrees C, indicating that percolation theory does not fully capture the dynamics of collagen network formation. The effects of collagen concentration and gelation temperature on k(g), t(c), and t(a) as well as on the mechanical properties and structure of these gels both during gelation and at equilibrium are elucidated. Composite collagen/HA gels are also prepared, and their properties are monitored at equilibrium and during gelation at 37 degrees C and 32 degrees C. We show that addition of HA subtly alters mechanical properties and structure of these systems both during the gelation process and at equilibrium. This occurs in a temperature-dependent manner, with the ratio of HA deposited on collagen fibers versus that distributed homogeneously between fibers increasing with decreasing gelation temperature. In addition to providing information on collagen and collagen/HA structure and mechanical properties during gelation, this work shows new ways in which rheology and microscopy can be used complementarily to reveal details of gelation processes. PMID- 19217875 TI - Ion trapping with fast-response ion-selective microelectrodes enhances detection of extracellular ion channel gradients. AB - Previously, functional mapping of channels has been achieved by measuring the passage of net charge and of specific ions with electrophysiological and intracellular fluorescence imaging techniques. However, functional mapping of ion channels using extracellular ion-selective microelectrodes has distinct advantages over the former methods. We have developed this method through measurement of extracellular K+ gradients caused by efflux through Ca2+-activated K+ channels expressed in Chinese hamster ovary cells. We report that electrodes constructed with short columns of a mechanically stable K+-selective liquid membrane respond quickly and measure changes in local [K+] consistent with a diffusion model. When used in close proximity to the plasma membrane (<4 microm), the ISMs pose a barrier to simple diffusion, creating an ion trap. The ion trap amplifies the local change in [K+] without dramatically changing the rise or fall time of the [K+] profile. Measurement of extracellular K+ gradients from activated rSlo channels shows that rapid events, 10-55 ms, can be characterized. This method provides a noninvasive means for functional mapping of channel location and density as well as for characterizing the properties of ion channels in the plasma membrane. PMID- 19217874 TI - Exploring local flexibility/rigidity in psychrophilic and mesophilic carbonic anhydrases. AB - Molecular flexibility and rigidity are required to determine the function and specificity of protein molecules. Some psychrophilic enzymes demonstrate a higher catalytic efficiency at low temperatures, compared to the efficiency demonstrated by their meso/thermophilic homologous. The emerging picture suggests that such enzymes have an improved flexibility of the structural catalytic components, whereas other protein regions far from functional sites may be even more rigid than those of their mesophilic counterparts. To gain a deeper insight in the analysis of the activity-flexibility/rigidity relationship in protein structure, psychrophilic carbonic anhydrase of the Antarctic teleost Chionodraco hamatus has been compared with carbonic anhydrase II of Bos taurus through fluorescence studies, three-dimensional modeling, and activity analyses. Data demonstrated that the cold-adapted enzyme exhibits an increased catalytic efficiency at low and moderate temperatures and, more interestingly, a local flexibility in the region that controls the correct folding of the catalytic architecture, as well as a rigidity in the hydrophobic core. The opposite result was observed in the mesophilic counterpart. These results suggest a clear relationship between the activity and the presence of flexible and rigid protein substructures that may be useful in rational molecular and drug design of a class of enzymes playing a key role in pathologic processes. PMID- 19217876 TI - Axisymmetric drop shape analysis for estimating the surface tension of cell aggregates by centrifugation. AB - Biological tissues behave in certain respects like liquids. Consequently, the surface tension concept can be used to explain aspects of the in vitro and in vivo behavior of multicellular aggregates. Unfortunately, conventional methods of surface tension measurement cannot be readily applied to small cell aggregates. This difficulty can be overcome by an experimentally straightforward method consisting of centrifugation followed by axisymmetric drop shape analysis (ADSA). Since the aggregates typically show roughness, standard ADSA cannot be applied and we introduce a novel numerical method called ADSA-IP (ADSA for imperfect profile) for this purpose. To examine the new methodology, embryonic tissues from the gastrula of the frog, Xenopus laevis, deformed in the centrifuge are used. It is confirmed that surface tension measurements are independent of centrifugal force and aggregate size. Surface tension is measured for ectodermal cells in four sample batches, and varies between 1.1 and 7.7 mJ/m2. Surface tension is also measured for aggregates of cells expressing cytoplasmically truncated EP/C cadherin, and is approximately half as large. In parallel, such aggregates show a reduction in convergent extension-driven elongation after activin treatment, reflecting diminished intercellular cohesion. PMID- 19217877 TI - Mitotic chromosome structure: reproducibility of folding and symmetry between sister chromatids. AB - Mitotic chromosome structure and pathways of mitotic condensation remain unknown. The limited amount of structural data on mitotic chromosome structure makes it impossible to distinguish between several mutually conflicting models. Here we used a Chinese hamster ovary cell line with three different lac operator-tagged vector insertions distributed over an approximately 1 microm chromosome arm region to determine positioning reproducibility, long-range correlation in large scale chromatin folding, and sister chromatid symmetry in minimally perturbed, metaphase chromosomes. The three-dimensional positions of these lac operator tagged spots, stained with lac repressor, were measured in isolated metaphase chromosomes relative to the central chromatid axes labeled with antibodies to topoisomerase II. Longitudinal, but not axial, positioning of spots was reproducible but showed intrinsic variability, up to approximately 300 nm, between sister chromatids. Spot positions on the same chromatid were uncorrelated, and no correlation or symmetry between the positions of corresponding spots on sister chromatids was detectable, showing the absence of highly ordered, long-range chromatin folding over tens of mega-basepairs. Our observations are in agreement with the absence of any regular, reproducible helical, last level of chromosome folding, but remain consistent with any hierarchical folding model in which irregularity in folding exists at one or multiple levels. PMID- 19217878 TI - Impact of actin rearrangement and degranulation on the membrane structure of primary mast cells: a combined atomic force and laser scanning confocal microscopy investigation. AB - Degranulation of bone marrow-derived mast cells (BMMCs) triggered by antigens (e.g., 2,4-dinitrophenylated bovine serum albumin (DNP-BSA) and secretagogues (e.g., poly-L-lysine) was investigated by combined atomic force microscopy (AFM) and laser scanning confocal microscopy (LSCM). This combination enables the simultaneous visualization and correlation of membrane morphology with cytoskeletal actin arrangement and intracellular granules. Two degranulation mechanisms and detailed membrane structures that directly corresponded to the two stimuli were revealed. In DNP-BSA triggered activation, characteristic membrane ridges formed in accordance with the rearrangement of underlying F-actin networks. Individual granules were visualized after they released their contents, indicating a "kiss-and-run" pathway. In BMMCs stimulated by poly-L-lysine, lamellopodia and filopodia were observed in association with the F-actin assemblies at and near the cell periphery, whereas craters were observed on the central membrane lacking F-actin. These craters represent a new membrane feature resulting from the "kiss-and-merge" granule fusion. This work provides what we believe is important new insight into the local membrane structures in correlation with the cytoskeleton arrangement and detailed degranulation processes. PMID- 19217880 TI - Mechanogenetic coupling of Hydra symmetry breaking and driven Turing instability model. AB - The freshwater polyp Hydra can regenerate from tissue fragments or random cell aggregates. We show that the axis-defining step ("symmetry breaking") of regeneration requires mechanical inflation-collapse oscillations of the initial cell ball. We present experimental evidence that axis definition is retarded if these oscillations are slowed down mechanically. When biochemical signaling related to axis formation is perturbed, the oscillation phase is extended and axis formation is retarded as well. We suggest that mechanical oscillations play a triggering role in axis definition. We extend earlier reaction-diffusion models for Hydra regrowth by coupling morphogen transport to mechanical stress caused by the oscillations. The modified reaction-diffusion model reproduces well two important experimental observations: 1), the existence of an optimum size for regeneration, and 2), the dependence of the symmetry breaking time on the properties of the mechanical oscillations. PMID- 19217879 TI - Cardiac myocyte excitation by ultrashort high-field pulses. AB - In unexcitable, noncardiac cells, ultrashort (nanosecond) high-voltage (megavolt per-meter) pulsed electrical fields (nsPEF) can mobilize intracellular Ca2+ and create transient nanopores in the plasmalemma. We studied Ca2+ responses to nsPEF in cardiac cells. Fluorescent Ca2+ or voltage signals were recorded from isolated adult rat ventricular myocytes deposited in an electrode microchamber and stimulated with conventional pulses (CPs; 0.5-2.4 kV/cm, 1 ms) or nsPEF (10-80 kV/cm, 4 ns). nsPEF induced Ca2+ transients in 68/104 cells. Repeating nsPEF increased the likelihood of Ca2+ transient induction (61.8% for <10 nsPEF vs. 80.6% for > or =10 nsPEF). Repetitive Ca2+ waves arising at the anodal side and Ca2+ destabilization occurred after repeated nsPEF (12/29) or during steady-state single nsPEF delivery at 2 Hz. Removing extracellular Ca2+ abolished responses to nsPEF. Verapamil did not affect nsPEF-induced Ca2+ transients, but decreased responses to CP. Tetrodotoxin and KB-R7943 increased the repetition threshold in response to nsPEF: 1-20 nsPEF caused local anodal Ca2+ waves without Ca2+ transients, and > or =20 nsPEF caused normal transients. Ryanodine-thapsigargin and caffeine protected against nsPEF-induced Ca2+ waves and showed less recovery of diastolic Ca2+ levels than CP. Voltage recordings demonstrated action potentials triggered by nsPEF, even in the presence of tetrodotoxin. nsPEF can mobilize intracellular Ca2+ in cardiac myocytes by inducing action potentials. Anodal Ca2+ waves and resistance to Na+ and Ca2+ channel blockade suggest nonselective ion channel transport via sarcolemmal nanopores as a triggering mechanism. PMID- 19217881 TI - The regulative role of neurite mechanical tension in network development. AB - A bewildering series of dynamical processes take part in the development of the nervous system. Neuron branching dynamics, the continuous formation and elimination of neural interconnections, are instrumental in constructing distinct neuronal networks, which are the functional building blocks of the nervous system. In this study, we investigate and validate the important regulative role of mechanical tension in determining the final morphology of neuronal networks. To single out the mechanical effect, we cultured relatively large invertebrate neurons on clean quartz surfaces. Applied to these surfaces were isolated anchoring sites consisting of carbon nanotube islands to which the cells and the neurites could mechanically attach. Inspection of branching dynamics and network wiring upon development revealed an innate selection mechanism in which one axon branch wins over another. The apparent mechanism entails the build-up of mechanical tension in developing axons. The tension is maintained by the attachment of the growth cone to the substrate or, alternatively, to the neurites of a target neuron. The induced tension promotes the stabilization of one set of axon branches while causing retraction or elimination of axon collaterals. We suggest that these findings represent a crucial, early step that precedes the formation of synapses and regulates neuronal interconnections. Mechanical tension serves as a signal for survival of the axonal branch and perhaps for the subsequent formation of synapses. PMID- 19217882 TI - Signal transduction pathways that regulate sperm capacitation and the acrosome reaction. AB - Ejaculated spermatozoa must undergo physiological priming as they traverse the female reproductive tract before they can bind to the egg's extracellular coat, the zona pellucida (ZP), undergo the acrosome reaction, and fertilize the egg. The preparatory changes are the net result of a series of biochemical and functional modifications collectively referred to as capacitation. Accumulated evidence suggests that the event that initiates capacitation is the efflux of cholesterol from the sperm plasma membrane (PM). The efflux increases permeability and fluidity of the sperm PM and causes influx of Ca(2+) ions that starts a signaling cascade and result in sperm capacitation. The binding of capacitated spermatozoa to ZP further elevates intrasperm Ca(2+) and starts a new signaling cascade which open up Ca(2+) channels in the sperm PM and outer acrosomal membrane (OAM) and cause the sperm to undergo acrosomal exocytosis. The hydrolytic action of the acrosomal enzymes released at the site of sperm-egg (zona) binding, along with the hyperactivated beat pattern of the bound spermatozoon, are important factors in directing the sperm to penetrate the ZP and fertilize the egg. The role of Ca(2+)-signaling in sperm capacitation and induction of the acrosome reaction (acrosomal exocytosis) has been of wide interest. However, the precise mechanism(s) of its action remains elusive. In this article, we intend to highlight data from this and other laboratories on Ca(2+) signaling cascades that regulate sperm functions. PMID- 19217883 TI - Knockdown of alphaII spectrin in normal human cells by siRNA leads to chromosomal instability and decreased DNA interstrand cross-link repair. AB - Nonerythroid alpha-spectrin (alphaIISp) is a structural protein involved in repair of DNA interstrand cross-links and is deficient in cells from patients with Fanconi anemia (FA), which are defective in ability to repair cross-links. In order to further demonstrate the importance of the role that alphaIISp plays in normal human cells and in the repair defect in FA, alphaIISp was knocked down in normal cells using siRNA. Depletion of alphaIISp in normal cells by siRNA resulted in chromosomal instability and cellular hypersensitivity to DNA interstrand cross-linking agents. An increased number of chromosomal aberrations were observed and, following treatment with a DNA interstrand cross-linking agent, mitomycin C, cells showed decreased cell growth and survival and decreased formation of damage-induced alphaIISp and XPF nuclear foci. Thus depletion of alphaIISp in normal cells leads to a number of defects observed in FA cells, such as chromosome instability and a deficiency in cross-link repair. PMID- 19217884 TI - Cloning and characterization of the 5'-flanking region of the pig adiponectin gene. AB - Adiponectin, a cytokine hormone secreted exclusively by adipose tissue, has key roles in energy homeostasis, and in glucose and lipid metabolism. To understand the regulatory expression of pig adiponectin, the 5'-flanking region of the adiponectin gene was isolated from a pig BAC library. 5'-RACE analysis revealed that there were three transcriptional start sites, including one that is novel. The luciferase reporter assay detected a positive cis-acting element for efficient expression of the adiponectin gene at the region spanned by nucleotides -1150 to -1130 with serially deleted 5'-flanking sequences as its promoters. Analysis of oligonucleotide competition by the electrophoretic mobility-shift assay revealed the presence of a cAMP-responsive element (CRE) (nucleotides -1150 to -1130) for the cAMP-responsive element binding protein (CREB), which has not been reported in human or mouse adiponectin genes. These results indicated that CREB is an essential regulatory factor for the transcriptional activity of pig adiponectin. PMID- 19217885 TI - A novel method of mouse ex utero transplantation of hepatic progenitor cells into the fetal liver. AB - Avoiding the limitations of the adult liver niche, transplantation of hepatic stem/progenitor cells into fetal liver is desirable to analyze immature cells in a hepatic developmental environment. Here, we established a new monitor tool for cell fate of hepatic progenitor cells transplanted into the mouse fetal liver by using ex utero surgery. When embryonic day (ED) 14.5 hepatoblasts were injected into the ED14.5 fetal liver, the transplanted cells expressed albumin abundantly or alpha-fetoprotein weakly, and contained glycogen in the neonatal liver, indicating that transplanted hepatoblasts can proliferate and differentiate in concord with surrounding recipient parenchymal cells. The transplanted cells became mature in the liver of 6-week-old mice. Furthermore, this method was applicable to transplantation of hepatoblast-like cells derived from mouse embryonic stem cells. These data indicate that this unique technique will provide a new in vivo experimental system for studying cell fate of hepatic stem/progenitor cells and liver organogenesis. PMID- 19217886 TI - cAMP enhances BMP2-signaling through PKA and MKP1-dependent mechanisms. AB - Recent studies suggest that the elevation of intracellular cyclic adenosine monophosphate (cAMP) and the activation of the protein kinase A regulate BMP induced osteogenesis. However, the precise mechanisms underlying the enhancing effect of cAMP on BMP2 signaling were not completely revealed. In this study we investigated the effect of elevated cAMP level and PKA activation on the BMP2 induced osteoblastic differentiation in pluripotent C2C12 cells. Alkaline phosphatase activity and its mRNA were consistently induced by BMP2 treatment. The pretreatment of C2C12 cells with Forskolin, a cAMP generating agent, dbcAMP, an analogue of cAMP, or IBMX (3-isobutyl 1-methyl xanthine), and a nonspecific inhibitor of phosphodiesterases elicited further activation of alkaline phosphatase. Furthermore, elevated intracellular cAMP level increased BMP2 induced MKP1. On the other hand, BMP2-induced Erk phosphorylation (p44/p42) and cell proliferation were suppressed in the presence of cAMP. Thus, cAMP might enhance BMP2-induced osteoblastic differentiation by a MKP1-Erk-dependent mechanism. PMID- 19217887 TI - Angiopoietins contribute to lung development by regulating pulmonary vascular network formation. AB - Angiopoietin (Ang) signaling through the Tie2 receptor regulates vasculature. The role of Ang signaling in pulmonary hypertension is well investigated, but its role in lung development is not elucidated. Here, we show that the Tie2 agonist ligand, Ang1, was detected in lung tissue at birth and its expression gradually increased in mice, whereas its antagonist Ang2 was abundant at birth and decreased inversely with Ang1. Mice expressing the potent chimeric Ang1 protein COMP-Ang1 in surfactant protein C (SPC)-positive lung epithelial cells, showed 50% lethality at birth due to respiratory failure. Surviving mice displayed impaired adaptive responsive respiratory function. Histological analysis revealed that pulmonary artery and alveolar structure were significantly dilated, and alveolar density was decreased to approximately a third of controls. Thus, the precise regulation of Tie2 signaling through an Ang1/Ang2 expression switch is important to construct a mature lung vascular network system required for normal lung development. PMID- 19217888 TI - Association of shorter mean telomere length with risk of incident myocardial infarction: a prospective, nested case-control approach. AB - BACKGROUND: Recent data have implicated telomere length shortening as a potential risk predictor for cardiovascular disease. However, to date, prospective epidemiological data are scarce. METHODS: Using leukocyte DNA samples collected at baseline in a prospective cohort of 14,916 initially healthy American men, we examined the relationship of mean telomere repeat copy number to single gene copy number (T/S ratio), using a re-modified quantitative polymerase chain reaction protocol, among 337 white males who subsequently developed an incident myocardial infarction (MI), and among an equal number of age- and smoking-matched white males who remained free of reported vascular disease during follow-up (controls). RESULTS: The mean follow-up time since randomization was 3.85 y. The T/S ratio was inversely correlated with age in the controls (R=-0.114; p=0.036). The log(e) transformed T/S ratios were significantly smaller in the MI cases (3.41+/-0.63) than the MI controls (3.52+/-0.78) (p=0.01). In a multi-variable adjusted analysis, decreased T/S ratio was significantly associated with risk of MI (odds ratio=1.621; 95%CI=1.140-2.304; p=0.007). CONCLUSIONS: The present investigation has shown an association of telomere length shortening with increased risk of incident myocardial infarction, further suggesting the importance of telomere biology in atherogenesis. PMID- 19217889 TI - Correlation of plasma and synovial fluid osteopontin with disease severity in knee osteoarthritis. AB - OBJECTIVES: The purposes of this study were to examine osteopontin levels in both plasma and synovial fluid of patients with primary knee osteoarthritis (OA) and to investigate their relationship with severity of the disease. DESIGN AND METHODS: Thirty-two patients aged 53-83 years with knee OA and 15 healthy controls were enrolled in this study. Anteroposterior knee radiographs were taken to determine the disease severity of the affected knee. The radiographic grading of OA in the knee was performed by using the Kellgren-Lawrence criteria. Osteopontin levels in the plasma and synovial fluid were measured using enzyme linked immunosorbent assay. RESULTS: The mean plasma osteopontin concentration of the knee OA patients was significantly higher compared with that of healthy controls (168.8+/-15.6 vs 67.2+/-7.7 ng/mL, P<0.0001). Osteopontin levels in synovial fluid were significantly higher with respect to paired plasma samples (272.1+/-15.0 vs 168.8+/-15.6 ng/mL, P<0.001). In addition, plasma osteopontin levels showed a positive correlation with synovial fluid osteopontin levels (r=0.373, P=0.035). Subsequent analysis showed that plasma osteopontin levels significantly correlated with severity of disease (r=0.592, P<0.001). Furthermore, the synovial fluid levels of osteopontin also correlated with disease severity (r=0.451, P=0.01). CONCLUSION: The data suggest that osteopontin in plasma and synovial fluid is related to progressive joint damage in knee OA. Osteopontin may serve as a biochemical marker for determining disease severity and could be predictive of prognosis with respect to the progression of knee OA. PMID- 19217890 TI - Plasma and urine betaine and dimethylglycine variation in healthy young male subjects. AB - OBJECTIVES: We aimed to compare the individuality (within subject consistency) of plasma and urine betaine and N,N-dimethylglycine. DESIGN AND METHODS: In two separate groups of 8 males (ages 19 to 40), plasma (10) and urine (6) samples were collected either over a single day or over an 8 week period. The individuality of the betaine and N,N-dimethylglycine plasma concentrations and excretions were estimated by one-way repeated measures analysis of variance. The reliability coefficients and indices of individuality were calculated. The between-subject variation in the study population was compared with that in a normal population (n=192 for plasma, 205 for urine). RESULTS: Plasma betaine concentrations were significantly different between subjects over 24 h and 8 weeks (p<0.00001). Plasma dimethylglycine concentrations were different over 24 h. Urine betaine and dimethylglycine excretions were different in both (p<0.0001). Betaine was more individual than dimethylglycine in both plasma and urine. Compared with a normal healthy population, the between-subject variation in plasma betaine was less (p<0.001) in the study group, but similar for dimethylglycine and for urine betaine. CONCLUSIONS: Plasma betaine and urinary betaine excretions are more individual than dimethylglycine. Plasma and urine betaine are highly individual in the general population. PMID- 19217891 TI - Decreased plasma levels of soluble receptor for advanced glycation endproducts (sRAGE) in patients with nonalcoholic fatty liver disease. AB - OBJECTIVES: Levels of soluble receptor for advanced glycation endproducts (sRAGE) have been linked to several components of the metabolic syndrome. We tested the hypothesis that plasma levels of sRAGE may be associated with non-alcoholic fatty liver disease. DESIGN AND METHODS: We enrolled subjects with definite nonalcoholic steatohepatitis (NASH, n=40), borderline NASH (n=8), simple fatty liver (n=9) and healthy controls (n=14). Plasma levels of sRAGE were measured by ELISA. RESULTS: Concentrations of sRAGE were significantly lower in patients with definite NASH (1080+/-392 pg/mL, P<0.01) and borderline NASH (1050+/-278 pg/mL, P<0.05) compared to controls (1480+/-387 pg/mL). Levels of sRAGE were significantly and inversely correlated with ALT (r=-0.30, P<0.05) and AST (r= 0.23, P<0.05). CONCLUSION: Plasma levels of sRAGE are significantly reduced in definite and borderline NASH. PMID- 19217892 TI - The deubiquitinase emperor's thumb is a regulator of apoptosis in Drosophila. AB - We have characterized the gene emperor's thumb (et) and showed that it is required for the regulation of apoptosis in Drosophila. Loss-of-function mutations in et result in apoptosis associated with a decrease in the concentration of DIAP1. Overexpression of one form of et inhibits apoptosis, consistent with et having an anti-apoptotic function; however, overexpression of a second form of et induces apoptosis, indicating that the two forms of et may have competing functions. et encodes a protein deubiquitinase, suggesting it regulates apoptosis by controlling the stability of apoptotic regulatory proteins. PMID- 19217893 TI - Temporal and spatial windows delimit activation of the outer ring of wingless in the Drosophila wing. AB - Extracellular signalling molecules play many roles in the development of higher organisms. They are used reiteratively in different tissues and stages, but the response of the receiving cells is controlled in a context dependent manner. The pattern of expression of the signalling molecule Wingless/WNT in Drosophila is extraordinarily complex. We have studied the mechanism that controls its expression and function in the outer ring of the Drosophila wing hinge. Our findings indicate that wingless expression is controlled by a dual mechanism: its initial activation requires the product of zinc finger homeodomain 2 and is subsequently repressed by the product of the gene complex elbow/no ocelli. This tight regulation restricts the activation of wingless temporally and spatially. Later in development, wingless expression is maintained by an autoregulatory loop that involves the product of homothorax. We have analyzed the phenotype of a wingless allelic combination that specifically removes the outer ring, and our results show that Wingless is required to promote local proliferation of the wing base cells. Thus, cell proliferation in the proximal-distal axis is controlled by the sequential activation of wingless in the inner ring and the outer ring at different stages of development. PMID- 19217894 TI - Bicaudal C and trailer hitch have similar roles in gurken mRNA localization and cytoskeletal organization. AB - Bicaudal C and trailer hitch are both required for dorsoventral patterning of the Drosophila oocyte. Each mutant produces ventralized eggs, a phenotype typically associated with failure of the oocyte to provide a dorsalization signal--the Gurken protein--to the follicle cells. Bicaudal C and trailer hitch are both implicated in post-transcriptional gene regulation. Bicaudal C acts in recruiting a deadenylase to specific mRNAs, leading to translational repression. The role of trailer hitch is less well defined, but mutants have defects in protein secretion, and show aberrant distribution of an endoplasmic reticulum exit site marker whose mRNA is associated with Trailer hitch protein. We show that Bicaudal C and trailer hitch interact genetically. Mutants of these two genes have shared defects in localization of gurken and other anteriorly-localized mRNAs, as well as altered microtubule organization which may underlie the mRNA localization defects. Bicaudal C and trailer hitch mutants also share a syndrome of actin related abnormalities, including the formation of ectopic actin cages near the anterior of the oocyte. The cages sequester Gurken protein, blocking its secretion and thus interfering with signaling of the follicle cells to specify dorsal fate. PMID- 19217896 TI - Origins and control of the differentiation of inhibitory interneurons and glia in the cerebellum. AB - Cerebellar GABAergic interneurons and glia originate from progenitors that delaminate from the ventricular neuroepithelium and proliferate in the prospective white matter. Even though this population of progenitor cells is multipotent as a whole, clonal analysis indicates that different lineages are already separated during postnatal development and little is known about the mechanisms that regulate the specification and differentiation of these cerebellar types at earlier stages. Here, we investigate the role of Ascl1 in the development of inhibitory interneurons and glial cells in the cerebellum. This gene is expressed by maturing oligodendrocytes and GABAergic interneurons and is required for the production of appropriate quantities of these cells, which are severely reduced in Ascl1(-/-) mouse cerebella. Nevertheless, the two lineages are not related and the majority of oligodendrocytes populating the developing cerebellum actually derive from extracerebellar sources. Targeted electroporation of Ascl1-expression vectors to ventricular neuroepithelium progenitors enhances the production of interneurons and completely suppresses astrocytic differentiation, whereas loss of Ascl1 function has opposite effects on both cell types. Our results indicate that Ascl1 directs ventricular neuroepithelium progenitors towards inhibitory interneuron fate and restricts their ability to differentiate along the astroglial lineage. PMID- 19217895 TI - The evolutionarily conserved leprecan gene: its regulation by Brachyury and its role in the developing Ciona notochord. AB - In Ciona intestinalis, leprecan was identified as a target of the notochord specific transcription factor Ciona Brachyury (Ci-Bra) (Takahashi, H., Hotta, K., Erives, A., Di Gregorio, A., Zeller, R.W., Levine, M., Satoh, N., 1999. Brachyury downstream notochord differentiation in the ascidian embryo. Genes Dev. 13, 1519 1523). By screening approximately 14 kb of the Ci-leprecan locus for cis regulatory activity, we have identified a 581-bp minimal notochord-specific cis regulatory module (CRM) whose activity depends upon T-box binding sites located at the 3'-end of its sequence. These sites are specifically bound in vitro by a GST-Ci-Bra fusion protein, and mutations that abolish binding in vitro result in loss or decrease of regulatory activity in vivo. Serial deletions of the 581-bp notochord CRM revealed that this sequence is also able to direct expression in muscle cells through the same T-box sites that are utilized by Ci-Bra in the notochord, which are also bound in vitro by the muscle-specific T-box activators Ci-Tbx6b and Ci-Tbx6c. Additionally, we created plasmids aimed to interfere with the function of Ci-leprecan and categorized the resulting phenotypes, which consist of variable dislocations of notochord cells along the anterior-posterior axis. Together, these observations provide mechanistic insights generally applicable to T-box transcription factors and their target sequences, as well as a first set of clues on the function of Leprecan in early chordate development. PMID- 19217897 TI - Perichondrial expression of Wdr5 regulates chondrocyte proliferation and differentiation. AB - Wdr5 is developmentally expressed in osteoblasts and is required for osteoblast differentiation. Mice overexpressing Wdr5 under the control of the mouse alpha(1)I collagen promoter (Col I-Wdr5) display accelerated osteoblast differentiation as well as accelerated chondrocyte differentiation, suggesting that overexpression of Wdr5 in osteoblasts affects chondrocyte differentiation. To elucidate the molecular mechanism by which overexpression of Wdr5 in the perichondrium regulates chondrocyte differentiation, studies were undertaken using skeletal elements and cultured metatarsals isolated from wild-type and Col I-Wdr5 embryos. FGF18 mRNA levels were decreased in Col I-Wdr5 humeri. Furthermore, local delivery of FGF18 to the bone collar of ex vivo cultures of metatarsals attenuated the chondrocyte phenotype of the Col I-Wdr5 metatarsals. Impairing local FGF action in wild-type metatarsals resulted in a chondrocyte phenotype analogous to that of Col I-Wdr5 metatarsals implicating impaired FGF action as the cause of the phenotype observed. The expression of Twist-1, which regulates chondrocyte differentiation, was increased in Col I-Wdr5 humeri. Chromatin immunoprecipitation analyses demonstrated that Wdr5 is recruited to the Twist-1 promoter. These findings support a model in which overexpression of Wdr5 in the perichondrium promotes chondrocyte differentiation by modulating the expression of Twist-1 and FGF18. PMID- 19217898 TI - Multiple Wnts are involved in Hydra organizer formation and regeneration. AB - Wnt genes and beta-catenin signaling are involved in axial patterning processes in vertebrate embryogenesis in setting up the Spemann-Mangold organizer in amphibian embryos. An organizer with a similar function is present in the hypostome of an adult Hydra polyp. Previously, a Hydra ortholog of Wnt3 (HyWnt3), which is expressed in the hypostome, has been described. Here, ten additional Hydra Wnt genes have been identified. Of these, six (HyWnt1, -7, -9/10a, -9/10c, 11, and -16) are expressed in the adult hypostome. And, as is HyWnt3, these six Wnt genes are also expressed when a new head organizer is formed during head regeneration and bud formation. The kinetics of Wnt gene expressions during head regeneration suggests that a cascade of consecutive Wnt activation accompanies regeneration, and HyWnt3 begins this cascade. Recombinant HyWnt3 protein induced body column tissue to undergo head formation. It also increased the head formation capacity in the head regeneration-deficient mutant strain reg-16 to that of wild-type strains. In addition our data reveal striking similarities in the molecular basis of the organizer in Hydra and axis polarization in chordates (e.g. Spemann's organizer) as well as it's role in regeneration suggesting a conserved function of Wnt signaling in setting up this ancient metazoan signaling center. PMID- 19217900 TI - Receptor reserve-dependent properties of antipsychotics at human dopamine D2 receptors. AB - Aripiprazole is the first dopamine D(2)/D(3) receptor partial agonist approved for use in the treatment of psychiatric disorders including schizophrenia, bipolar disorder, and unipolar depression in the US. Aripiprazole has demonstrated a relatively favorable side effect profile compared to other commonly prescribed antipsychotics, including a low propensity for treatment limiting extrapyramidal symptoms, hyperprolactinemia, and body weight gain. In an effort to elucidate aripiprazole's pharmacological activity in relation to clinically relevant fluctuation of dopamine D(2) receptor reserves, we compared the properties of aripiprazole to other antipsychotics, quetiapine, clozapine, olanzapine, ziprasidone, risperidone and haloperidol, a dopamine D(2) receptor partial agonist, bifeprunox, dopamine D(3) receptor modulators, BP897 (N-[4-[4-(2 Methoxyphenyl)-1-piperazinyl]butyl]naphthalene-2-carboxamide) and GR103691 (4' Acetyl-N-[4-[4-(2-methoxyphenyl)piperazin-1-yl]butyl]biphenyl-4-carboxamide), and a 5-HT(1A) partial agonist, buspirone using forskolin-stimulated cAMP accumulation in clonal Chinese hamster ovary cell lines expressing low and high densities of human dopamine D(2S) receptors (hD(2S)-Low and hD(2S)-High, respectively). In hD(2S)-Low cells lacking receptor reserves for dopamine, all drugs antagonized dopamine responses, and their potencies correlated well with respective affinities. In hD(2S)-High cells possessing receptor reserves, all antipsychotics except aripiprazole antagonized dopamine responses, and their antagonist potencies were less than those in hD(2S)-Low cells treated with the equal dopamine concentration. In contrast, aripiprazole and bifeprunox acted as full agonists. BP897, GR103691 and buspirone acted as partial agonists. These data suggest that the level of receptor reserves influences antagonist potencies and side effects associated with antipsychotics. Aripiprazole's unique receptor reserve dependent properties may account for its favorable tolerability in the clinical setting. PMID- 19217901 TI - Gastroprotective effect of simvastatin against indomethacin-induced gastric ulcer in rats: role of nitric oxide and prostaglandins. AB - This study investigated the possible mechanisms underlying the gastroprotective effect of simvastatin against indomethacin-induced gastric ulcer in rats. Rats were randomly assigned to vehicle-, simvastatin-, simvastatin+L-arginine- and simvastatin+N(G)-nitro-L-arginine methyl ester (L-NAME)-pretreated groups for two weeks. Pyloric ligation was performed for the collection of gastric juice, and gastric ulceration was induced by a single intraperitoneal injection of indomethacin (30 mg/kg). Gastric juice parameters (total acid output, pepsin activity and mucin concentration) were determined. The stomachs tissues were used for determination of gastric mucosal lipid peroxides, superoxide dismutase, catalase, total nitrites and prostaglandin E(2) levels. Pretreatment with simvastatin (10 mg/kg, orally, for 2 weeks) caused significant reduction in gastric mucosal lesions and lipid peroxides associated with a significant increase in gastric juice mucin concentration. Simvastatin significantly increased the gastric mucosal total nitrite and prostaglandin E(2) levels. Additionally, simvastatin attenuated the elevations in gastric mucosal superoxide dismutase observed with indomethacin. The gastroprotective effect afforded by simvastatin was significantly augmented by coadministration with L-arginine (a nitric oxide precursor) and inhibited by coadministration with L-NAME (a nitric oxide synthase inhibitor). Results confirm a gastroprotective effect for simvastatin, and indicate that the anti-ulcer effect of simvastatin is mediated by scavenging free radicals, increasing nitric oxide and prostaglandin E(2) levels, and increasing gastric juice mucin production. We conclude that simvastatin represents a more suitable antihyperlipidemic therapy for patients who are at risk of gastric ulcers that were induced by the use of nonsteroidal anti-inflammatory drugs (NSAIDs). PMID- 19217902 TI - Biomechanics of the optic nerve head. AB - Biomechanical factors acting at the level of the lamina cribrosa (LC) are postulated to play a role in retinal ganglion cell dysfunction and loss in glaucoma. In support of this postulate, we now know that a number of cell types (including lamina cribrosa cells) are mechanosensitive. Here we briefly review data indicating cellular stretching, rate of stretching and substrate stiffness may be important mechanosensitivity factors in glaucoma. We then describe how experiments and finite element modeling can be used to quantify the biomechanical environment within the LC, and how this environment depends on IOP. Generic and individual-specific models both suggest that peripapillary scleral properties have a strong influence on LC biomechanics, which can be explained by the observation that scleral deformation drives much of the IOP-dependent straining of the LC. Elegant reconstructions of the LC in monkey eyes have shown that local strains experienced by LC cells depend strongly on laminar beam microarchitecture, which can lead to large local strain elevations. Further modeling, suitably informed by experiments, is needed to better understand lamina cribrosa biomechanics and how they may be involved in glaucomatous optic neuropathy. PMID- 19217899 TI - Regulation of mouse embryonic stem cell neural differentiation by retinoic acid. AB - Pluripotent mouse embryonic stem cells (ESCs) derived from the early blastocyst can differentiate in vitro into a variety of somatic cell types including lineages from all three embryonic germ layers. Protocols for ES cell neural differentiation typically involve induction by retinoic acid (RA), or by exposure to growth factors or medium conditioned by other cell types. A serum-free differentiation (SFD) medium completely lacking exogenous retinoids was devised that allows for efficient conversion of aggregated mouse ESCs into neural precursors and immature neurons. Neural cells produced in this medium express neuronal ion channels, establish polarity, and form functional excitatory and inhibitory synapses. Brief exposure to RA during the period of cell aggregation speeds neuronal maturation and suppresses cell proliferation. Differentiation without RA yields neurons and neural progenitors with apparent telencephalic identity, whereas cells differentiated with exposure to RA express markers of hindbrain and spinal cord. Transcriptional profiling indicates a substantial representation of transit amplifying neuroblasts in SFD cultures not exposed to RA. PMID- 19217903 TI - G1961E mutant allele in the Stargardt disease gene ABCA4 causes bull's eye maculopathy. AB - The aim of this study was to characterize the pathological and functional consequences of the G1961E mutant allele in the Stargardt disease gene ABCA4. Data from 15 patients were retrospectively reviewed and all the patients had at least one G1961E mutation. Comprehensive ophthalmic examination, full-field and pattern electroretinograms, and fundus autofluorescence (FAF) imaging were performed on all patients. Microperimetry, spectral-domain optical coherence tomography (OCT), and fluorescein angiography were performed in selected cases. Genetic screening was performed using the ABCR400 micro-array that currently detects 496 distinct ABCA4 variants. All patients had normal full-field scotopic and photopic electroretinograms (ERGs) and abnormal pattern electroretinograms (PERGs) performed on both eyes, and all the fundi had bull's eye maculopathy without retinal flecks on FAF. On OCT, 1 patient had disorganization of photoreceptor outer segment, 2 had outer nuclear layer (ONL) thinning likely due to photoreceptor atrophy proximal to the foveal center, and 3 had additional retinal pigment epithelium (RPE) atrophy. On microperimetry, 6 patients had eccentric superior fixation and amongst this group, 5 had an absolute scotoma in the foveal area. DNA analysis revealed that 3 patients were homozygous G1961E/G1961E and the rest were compound heterozygotes for G1961E and other ABCA4 mutations. The G1961E allele in either homozygosity or heterozygosity is associated with anatomical and functional pathologies limited to the parafoveal region and a trend to delayed onset of symptoms, relative to other manifestations of ABCA4 mutations. Our observations support the hypothesis that the G1961E allele contributes to localized macular changes rather than generalized retinal dysfunction, and is a cause of bull's eye maculopathy in either the homozygosity or heterozygosity state. In addition, genetic testing provides precise diagnosis of the underlying maculopathy, and current non-invasive imaging techniques could be used to detect photoreceptor damage at the earliest clinical onset of the disease. PMID- 19217905 TI - The developing mammalian retina is partially protected from gentamicin toxicity. AB - Gentamicin retinal toxicity was tested in numerous studies on adult animal models, but not in the developing retina. Since differentiating cells and developing tissues may exhibit different degrees of sensitivity to toxic drugs, we aimed here to test the susceptibility of the developing mammalian retina to the toxic action of gentamicin. Gentamicin was injected in the right eye of newborn rabbits, aged 11-38 days. The left eye of each rabbit was injected with saline. Eight weeks after injection, electroretinographic (ERG) responses were recorded in the dark- and light-adapted states. The rabbits were then sacrificed, and the retinas were prepared for morphological examination and immunostaining for Glial Fibrillary Acidic Protein (GFAP) and Protein Kinase C (PKC). The ERG responses demonstrated practically no functional damage to the retina of rabbits injected at postnatal age of 11 and 13 days, and a gradually increasing severity of functional damage with increasing postnatal age of intravitreal gentamicin injection. The histopathological studies yielded similar results. GFAP immunoreactivity showed staining in Muller cells only in retinas that exhibited functional and structural damage. PKC immunoreactivity indicated lesser damage to the rod-bipolar cells compared to the photoreceptors. The ERG data and morphological observations suggest that processes involved in development of the rabbit retina, such as outer segment elongation may provide partial or even complete protection to the retina from toxic insults such as a single dose of gentamicin. PMID- 19217904 TI - Neurotrophin roles in retinal ganglion cell survival: lessons from rat glaucoma models. AB - The neurotrophin (NT) hypothesis proposes that the obstruction of retrograde transport at the optic nerve head results in the deprivation of neurotrophic support to retinal ganglion cells (RGC) leading to apoptotic cell death in glaucoma. An important corollary to this concept is the implication that appropriate enhancement of neurotrophic support will prolong the survival of injured RGC indefinitely. This hypothesis is, perhaps, the most widely recognized theory to explain RGC loss resulting from exposure of the eye to elevated intraocular pressure (IOP). Recent studies of NT signaling using rat glaucoma models, have examined the endogenous responses of the retina to pressure exposure as well as studies designed to augment NT signaling in order to rescue RGC from apoptosis following pressure-induced injury. The examination of these studies in this review reveals a number of consistent observations and provides direction for further investigations of this hypothesis. PMID- 19217906 TI - Trypanosoma cruzi: isolation and characterization of aspartyl proteases. AB - Two aspartyl proteases activities were identified and isolated from Trypanosoma cruzi epimastigotes: cruzipsin-I (CZP-I) and cruzipsin-II (CZP-II). One was isolated from a soluble fraction (CZP-II) and the other was solubilized with 3 [(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CZP-I). The molecular mass of both proteases was estimated to be 120 kDa by HPLC gel filtration and the activity of the enzymes was detected in a doublet of bands (56 and 48 kDa) by substrate-sodium dodecyl sulphate-polyacrylamide-gelatin gel electrophoresis. Substrate specificity studies indicated that the enzymes consistently hydrolyze the cathepsin D substrate Phe-Ala-Ala-Phe (4-NO2)-Phe-Val-Leu-O4MP but failed to hydrolyze serine and other protease substrates. Both proteases activities were strongly inhibited by the classic inhibitor pepstatin-A (> or =68%) and the aspartic active site labeling agent, 1,2-epoxy-3-(phenyl-nitrophenoxy) propane (> or =80%). These findings show that both proteases are novel T. cruzi acidic proteases. The physiological function of these enzymes in T. cruzi has under investigation. PMID- 19217907 TI - Toxoplasma gondii: congenital transmission in a hamster model. AB - The objective of the research was to test the hamster for a model of transmission of congenital toxoplasmosis. A non-invasive method for the diagnosis of pregnancy in hamsters was designed, with a specificity and a sensitivity of 70.2 and 94.7%, respectively (n=168). Of 32 females with a chronic toxoplasma infection, 3 transmitted Toxoplasma congenitally during their first pregnancy, but not during the subsequent pregnancy. Congenital transmission rates of infections initiated during pregnancy with 2 stages of 2 strains of Toxoplasma were in the range of 33 to 100% of the 76 females inoculated. Only 1 of 17 females transmitted the parasite exclusively via milk. It was concluded that the hamster is a promising species for a model of transmission of congenital toxoplasmosis. PMID- 19217908 TI - Toxoplasmosis: A history of clinical observations. AB - It has been 100 years since Toxoplasma gondii was initially described in Tunis by Nicolle and Manceaux (1908) in the tissues of the gundi (Ctenodoactylus gundi) and in Brazil by Splendore (1908) in the tissues of a rabbit. Toxoplasma gondii is a ubiquitous, Apicomplexan parasite of warm-blooded animals that can cause several clinical syndromes including encephalitis, chorioretinitis, congenital infection and neonatal mortality. Fifteen years after the description of T. gondii by Nicolle and Manceaux a fatal case of toxoplasmosis in a child was reported by Janku. In 1939 Wolf, Cowen and Paige were the first to conclusively identify T. gondii as a cause of human disease. This review examines the clinical manifestations of infection with T. gondii and the history of the discovery of these manifestations. PMID- 19217910 TI - Type 2 diabetes, mitochondrial biology and the heart. AB - Diabetes is recognized as an independent risk factor for cardiovascular morbidity and mortality. This is due, in large part, to premature atherosclerosis, enhanced thrombogenicity and activation of systemic inflammatory programs with resultant vascular dysfunction. More enigmatic mechanisms underpinning diabetes-associated cardiac pathophysiology include the direct metabolic consequences of this disease on the myocardium. Nevertheless, a role for diabetes-associated disruption in cardiac contractile mechanics and in increasing cardiomyocyte susceptibility to ischemic-stress has been implicated independent of vascular pathology. This review will focus broadly on the direct effects of diabetes on the cardiac myocardium with more specific reference to the role of the modulation of cardiomyocyte mitochondrial function in these disease processes. This focus in part, stems from the growing recognition that in some instances mitochondrial dysfunction is central to the development of insulin resistance and diabetes, and in others, diabetes associated disruption in mitochondrial function exacerbates and accentuates the pathophysiology of diabetes. PMID- 19217911 TI - Downhill versus barrier-limited folding of BBL 3. Heterogeneity of the native state of the BBL peripheral subunit binding domain and its implications for folding mechanisms. AB - Protein folding studies are generally predicated on Anfinsen's dogma that there is a unique native state of a protein. However, this is not always the case. NMR measurements of BBL, for example, find a decrease in helicity of helix 2 surrounding His166 on its protonation, which, with other experimental data, suggests that the native state can occupy two or more conformations. Here, we analysed the native structure of BBL as a function of pH, temperature and ionic strength, along with a truncated BBL construct, by extensive all-atom molecular dynamics simulations in explicit solvent, corresponding to at least 400 ns of trajectories collected for each set of conditions. The native state was heterogeneous under a variety of conditions, consisting of two predominant conformations. This equilibrium changed with conditions: protonation of His166 at low pH shifted the equilibrium in favour of a less ordered conformer, while high ionic strength at neutral pH shifted the equilibrium to a more ordered conformer. Furthermore, high temperature and truncation of the sequence also shifted the equilibrium toward the less ordered conformer. Importantly, conformational heterogeneity in a native structure that changes with conditions will lead to deviations from the classic two-state behaviour during the barrier-limited unfolding of a protein. In particular, some regions of the protein will appear to unfold asynchronously and some residues will have anomalous thermal titration curves and unusual baseline behaviour monitored microscopically by NMR spectroscopy and macroscopically by calorimetry and other techniques. Such data could otherwise be interpreted as evidence for barrier-free downhill folding. Any biological significance of downhill folding of BBL appears to be ruled out by recent crystallographic studies on the reaction cycle of the BBL-equivalent domain in a pyruvate dehydrogenase multienzyme complex in which the domain remains of constant structure. PMID- 19217909 TI - Sexual recombination punctuated by outbreaks and clonal expansions predicts Toxoplasma gondii population genetics. AB - The cosmopolitan parasitic pathogen Toxoplasma gondii is capable of infecting essentially any warm-blooded vertebrate worldwide, including most birds and mammals, and establishes chronic infections in one-third of the globe's human population. The success of this highly prevalent zoonosis is largely the result of its ability to propagate both sexually and clonally. Frequent genetic exchanges via sexual recombination among extant parasite lineages that mix in the definitive felid host produces new lines that emerge to expand the parasite's host range and cause outbreaks. Highly successful lines spread clonally via carnivorism and in some cases sweep to pandemic levels. The extent to which sexual reproduction versus clonal expansion shapes Toxoplasma's current, global population genetic structure is the central question this review will attempt to answer. PMID- 19217913 TI - Comparative genomic analysis by microbial COGs self-attraction rate. AB - Whole genome analysis provides new perspectives to determine phylogenetic relationships among microorganisms. The availability of whole nucleotide sequences allows different levels of comparison among genomes by several approaches. In this work, self-attraction rates were considered for each cluster of orthologous groups of proteins (COGs) class in order to analyse gene aggregation levels in physical maps. Phylogenetic relationships among microorganisms were obtained by comparing self-attraction coefficients. Eighteen dimensional vectors were computed for a set of 168 completely sequenced microbial genomes (19 archea, 149 bacteria). The components of the vector represent the aggregation rate of the genes belonging to each of 18 COGs classes. Genes involved in nonessential functions or related to environmental conditions showed the highest aggregation rates. On the contrary genes involved in basic cellular tasks showed a more uniform distribution along the genome, except for translation genes. Self-attraction clustering approach allowed classification of Proteobacteria, Bacilli and other species belonging to Firmicutes. Rearrangement and Lateral Gene Transfer events may influence divergences from classical taxonomy. Each set of COG classes' aggregation values represents an intrinsic property of the microbial genome. This novel approach provides a new point of view for whole genome analysis and bacterial characterization. PMID- 19217914 TI - Tissue-dependent preventive effect of metallothionein against DNA damage in dyslipidemic mice under repeated stresses of fasting or restraint. AB - AIMS: To investigate the effect of repeated stress on DNA damage in seven organs of dyslipidemic mice, and the preventive role of metallothionein (MT). MAIN METHODS: Female adult 129/Sv wild-type and MT-null mice fed high-fat diet (HFD) were repeatedly subjected to mild stress of fasting or restraint in weeks 2 to 4 of 4-week study period. Serum cholesterol level, DNA damage in the liver, pancreas, spleen, bone marrow, kidney, lung and gastric mucosa, and other parameters were determined. KEY FINDINGS: Body weights were increased in both types of mice fed HFD compared to those fed standard diet (STD), and further increased by 12 h-fasting, while they were markedly decreased by 1-3 h-restraint. Fasting accelerated accumulation of fat in the liver, and increase in serum cholesterol of both types of mice fed HFD. Feeding of HFD increased DNA damage in the pancreas, spleen and bone marrow of both types of mice, compared with those fed STD. In the wild-type mice fed HFD, 24 h-fasting increased DNA damage in the liver and spleen, while restraint increased the damage in the liver, pancreas, spleen and bone marrow. DNA damage in the cells of organs was markedly increased in the MT-null mice. Specifically, damage in the liver, pancreas, spleen and bone marrow was greatly increased with the intensity of stress increased, and the damage was much greater in the restraint mice than in the fasting mice. SIGNIFICANCE: MT plays a tissue-dependent preventive role against DNA damage in various murine organs induced by repeated stress. PMID- 19217912 TI - Exploring the mechanism of beta-amyloid toxicity attenuation by multivalent sialic acid polymers through the use of mathematical models. AB - beta-Amyloid peptide (A beta), the primary protein component in senile plaques associated with Alzheimer's disease (AD), has been implicated in neurotoxicity associated with AD. Previous studies have shown that the A beta-neuronal membrane interaction plays a role in the mechanism of A beta toxicity. More specifically, it is thought that A beta interacts with ganglioside rich and sialic acid rich regions of cell surfaces. In light of such evidence, we have used a number of different sialic acid compounds of different valency or number of sialic acid moieties per molecule to attenuate A beta toxicity in a cell culture model. In this work, we proposed various mathematical models of A beta interaction with both the cell membrane and with the multivalent sialic acid compounds, designed to act as membrane mimics. These models allow us to explore the mechanism of action of this class of sialic acid membrane mimics in attenuating the toxicity of A beta. The mathematical models, when compared with experimental data, facilitate the discrimination between different modes of action of these materials. Understanding the mechanism of action of A beta toxicity inhibitors should provide insight into the design of the next generation of molecules that could be used to prevent A beta toxicity associated with AD. PMID- 19217915 TI - Caffeine alters proliferation of neuronal precursors in the adult hippocampus. AB - Neurogenesis continues through adulthood in the hippocampus and olfactory bulb of mammals. Adult neurogenesis has been implicated in learning and memory, and linked with depression. Hippocampal neurogenesis is increased in response to a number of stimuli, including exposure to an enriched environment, increased locomotor activity, and administration of antidepressants. Adult neurogenesis is depressed in response to aging, stress and sleep deprivation. Intriguingly, caffeine modulates a number of these same stimuli in a dose dependent manner. We examined the dose and duration dependent effects of caffeine on the proliferation, differentiation, and survival of newly generated hippocampal neurons in adult mice. Extended, 7 day caffeine administration, alters the proliferation of adult hippocampal precursors in the mouse in a dose dependent manner; moderate to high doses (20-30 mg/kg per day) of caffeine depress proliferation while supraphysiological doses (60 mg/kg per day) increase proliferation of neuronal precursors. Acute, 1 day administration had no affect on proliferation. Caffeine administration does not affect the expression of early or late markers of neuronal differentiation, or rates of long-term survival. However, neurons induced in response to supraphysiological levels of caffeine have a lower survival rate than control cells; increased proliferation does not yield an increase in long-term neurogenesis. These results demonstrate that physiologically relevant doses of caffeine can significantly depress adult hippocampal neurogenesis. PMID- 19217916 TI - Discussion of unique properties of botulinum toxins. PMID- 19217920 TI - Evaluating the mucoadhesive properties of drug delivery systems based on hydrated thiolated alginate. AB - Mucoadhesive polymers have been proposed as drug delivery carriers due to their ability to adhere to the mucus layer. A relatively new class of mucoadhesive polymers, termed thiomers, was suggested as an improved carrier capable of creating disulfide covalent bond with the mucus. Since the wet physiological environment is likely to cause any delivery system to adsorb water and arrive hydrated to its target, studying the performance of mucoadhesive systems in their hydrated form is of major importance. Model thiomer, alginate-thiol, were synthesized and characterized the product using Nuclear Magnetic Resonance (NMR), Fourier Transform Infra Red spectroscopy (FTIR). The swelling behavior was determined gravimetrically and found to be affected from the thiolation. Interactions between the alginate-thiol and mucin glycoproteins, which are believed to be an outcome of disulfide bonds, were verified using rheology experiments. Adhesion of hydrated tablets with different cross linking densities to porcine's fresh small intestine tissue were characterized using a Lloyd Tensile Machine. It was shown that the thiolation did not improve the adhesion properties of hydrated tablets. It appears that the benefit achieved by adding thiol group to the polymer in dry tablet form was flawed in hydrated form due to formation of inter-molecular disulfide junctions. PMID- 19217918 TI - Baclofen, raclopride, and naltrexone differentially affect intake of fat/sucrose mixtures under limited access conditions. AB - This study assessed the effects of the opioid antagonist naltrexone, the dopamine 2-like (D2) antagonist raclopride, and the GABA(B) agonist baclofen on consumption of fat/sucrose mixtures (FSM) using a limited access protocol. Sixty male Sprague-Dawley rats were grouped according to two schedules of access (Daily [D] or Intermittent [I]) to an optional FSM. Each FSM was created by whipping 3.2% (L), 10% (M), or 32% (H) powdered sugar into 100% vegetable shortening in a w/w manner (n=10 per group). One-hour intakes of the IL and IM groups were significantly greater than intakes of the respective DL and DM groups, thus fulfilling our operational definition of binge-type eating in these groups. Baclofen reduced intakes of the L and M mixtures regardless of access schedule, but failed to reduce intake of the H mixture. Naltrexone reduced intake in all groups, but potency was greater in IL rats than in DL rats. Furthermore, potency was attenuated in Intermittent rats, but enhanced in Daily rats, at higher sucrose concentrations. Raclopride reduced intake in the DL and stimulated intake in the IL groups, reduced intake in both M groups, and was without effect in both H groups. These results indicate that fat/sucrose mixtures containing relatively low concentrations of sucrose allow distinctions to be made between: 1) intakes stimulated by different access schedules and 2) opioid and dopaminergic modulation of those intakes. These results also suggest that brief bouts of food consumption involving fatty, sugar-rich foods may prove to be particularly resistant to pharmacological intervention. PMID- 19217919 TI - Intracellular signaling pathways involved in inhibition of PAI-1 expression by CNP in endothelial cells. AB - PAI-1 is a multifunctional protein stimulated by infectious agents and its activation is mediated by inflammatory cytokines such as TNFalpha. Recent studies demonstrate that natriuretic peptides, particularly C-type (CNP), can affect PAI 1 expression in bovine aortic smooth muscle cells and rat aortic endothelial cells. We have previously shown that CNP inhibits both basal and TNFalpha induced expression of PAI-1 in human endothelial cells. Herein, we describe mechanism by which CNP modulates signaling engaged in controlling PAI-1 expression in human endothelial cells. To examine which pathway initiated by TNFalpha is influenced, we tested kinase activity of MAP, PI3K/AKT and involvement of cGMP in endothelial cells exposed to CNP. CNP significantly increased cGMP level in endothelial cells. Its analogue, 8-Br-cGMP alone had no effect but significantly inhibited TNFalpha induced expression of PAI-1. Similarly, CNP and the inhibitors of ERK1/2 (PD098059) and PI3K (LY294002) attenuated PAI-1 expression induced by TNFalpha. CNP almost abolished TNFalpha induced phosphorylation of ERK1/2 but did not affect JNK phosphorylation, indicating that its effect on ERK1/2 was specific. These data suggest that CNP might function as the natural defense of vascular wall against cytokine induced PAI-1 release through its ability to inactivate PI3K/AKT and MEK/ERK pathways. PMID- 19217921 TI - Efficient polyethylenimine-mediated gene delivery proceeds via a caveolar pathway in HeLa cells. AB - Most in vivo gene therapies will require cell-specific targeting. Although vector targeting through ligand attachment has met with success in generating gene delivery particles that are capable of specific cellular interactions, little attention has been given to the possible effects of such ligands on subsequent intracellular processing. In this study, we examine the impact of targeting two distinct endocytic routes-the caveolar and clathrin pathways-on polyethylenimine mediated gene delivery in HeLa cells. Targeting complexes to the caveolar pathway with folic acid and the clathrin pathway with transferrin yields enhanced gene delivery relative to unmodified polyethylenimine. Colocalization studies with caveolin-1 and clathrin heavy chain indicate that the ligands successfully deliver their cargo to the intended pathways. However, inhibition of only the caveolar pathway-whether through the use of small molecule drugs or RNA interference-reduces gene delivery efficiency, suggesting that successful polyethylenimine-mediated gene delivery proceeds via a caveolar pathway in HeLa cells. Transfections in the presence of chloroquine and pH tracking studies reveal that a contributing factor to the success of the caveolar pathway is avoidance of lysosomes. Collectively, these data demonstrate that uptake mechanism and subsequent endocytic processing are important design parameters for gene delivery materials. PMID- 19217922 TI - Myeloid dendritic cell: From sentinel of immunity to key player of peripheral tolerance? AB - Myeloid dendritic cells (DC) are "sentinels" of immunity, ideally positioned throughout the body gateways and equipped with unique properties to transport antigens from the periphery to lymphoid tissues. They are professional antigen presenting cells transmitting incoming infectious signals to T cells, the key players of adaptive immunity. For induction of effective antigen-specific T-cell immunity, crosstalk of DC and naive T cells is mandatory. However, besides this essential immunostimulatory function of DC, consolidated findings from the DC research field in the last 10 years have shown that DC have an additional important function. They act as pivotal players in the peripheral tolerance network by active induction of T cells with immunosuppressive functions and regulation of T effector cell activity. This review summarizes the current knowledge about this immunoregulatory role of myeloid DC as guardians of T-cell immunity and T-cell tolerance and discusses the approaches of tolerogenic DC for therapeutic intervention strategies to diminish overwhelming immune responses in clinical application. PMID- 19217923 TI - A comparison of effects on reproduction and neonatal development in cynomolgus monkeys given human soluble IL-4R and mice given murine soluble IL-4R. AB - The effects of treatment with a soluble IL-4 receptor (sIL-4R) on reproduction and neonatal development were assessed in pregnant cynomolgus monkeys and mice. When pregnant cynomolgus monkeys were administered a human sIL-4R intravenously twice a week during organogenesis (GD 20-51) at 0, 0.2 or 2.0mg/kg, there was an increase in abortion/embryo-fetal death in the 0.2 (42.9%) and 2.0 (26.3%) mg/kg groups compared to controls (17.6%). All fetuses removed at cesarean sectioning on GD 100-102 were alive and no abnormalities were noted. There were three stillborn neonates (2.0mg/kg group), which were determined to have died before birth. No neonates died after birth and no abnormalities were noted. Due to the unanticipated results in the monkey study, a mouse developmental study with a murine surrogate molecule was conducted. When pregnant Crl:CD-1((R))(ICR)BR mice were administered murine sIL-4R intravenously once daily during the organogenesis period (GD 6-15) at 0, 25, 75, 250, or 625microg/mouse ( approximately 20mg/kg), there were no test-article-related abnormalities in any parameters. Antibody development to the drug did not influence toxicity in the monkey or mouse. In conclusion, evaluation of reproductive effects in mice administered murine soluble IL-4R was not predictive of reproductive effects noted in cynomolgus monkeys administered human soluble IL-4R. PMID- 19217924 TI - Dopaminergic hypofunctions and prepulse inhibition deficits in mice lacking midkine. AB - Midkine is a 13-kDa retinoic acid-induced heparin-binding growth factor involved in various biological phenomena such as cell migration, neurogenesis, and tissue repair. We previously demonstrated that midkine-deficient (Mdk(-/-)) mice exhibited a delayed hippocampal development with impaired working memory and increased anxiety only at the age of 4 weeks. To assess whether midkine gene could play important roles in development and maintenance of central nervous system, we investigated biochemical and behavioral parameters in dopamine and glutamate neurotransmission of Mdk(-/-) mice. The Mdk(-/-) mice exhibited a hypodopaminergic state (i.e., decreased levels of dopamine and its receptors in the striatum) with no alterations of glutamatergic system (i.e., normal level of glutamate, glutamine, glycine, d-serine, l-serine, and NMDA receptors in the frontal cortex and hippocampus). We also found prepulse inhibition deficits reversed by clozapine and haloperidol in the Mdk(-/-) mice. Our results suggested that midkine deficiency may be related to neurochemical and behavioral dysfunctions in dopaminergic system. PMID- 19217925 TI - A 28-day repeat dose toxicity study of steroidal glycoalkaloids, alpha-solanine and alpha-chaconine in the Syrian Golden hamster. AB - Glycoalkaloids alpha-solanine and alpha-chaconine are naturally present toxicants in the potato plant (Solanumtuberosum). Human intake of high doses of glycoalkaloids has led to acute intoxication, in severe cases coma and death. Previous studies have indicated that the ratio of alpha-solanine to alpha chaconine may determine the degree and nature of the glycoalkaloid toxicity in potatoes, as the toxicity of the two alkaloids act synergistically. The aim of the present study was to investigate whether an altered ratio of alpha-solanine and alpha-chaconine would reduce the toxicity of the glycoalkaloids. The Syrian Golden hamster was given daily doses of alpha-solanine and alpha-chaconine by gavage for 28 days. Doses of up to 33.3 mg total glycoalkaloids/kg body weight were applied in ratios of 1:3.7 and 1:70 (alpha-solanine:alpha-chaconine). Administration of the highest doses of both ratios resulted in distended and fluid filled small intestines and stomach. Animals receiving the ratio with the reduced content of alpha-solanine were less affected compared to those receiving the other ratio. Gene expression profiling experiments were conducted using RNA from epithelial scrapings from the small intestines of the hamsters administered the highest doses of the glycoalkaloid treatments. In general, more differential gene expression was observed in the epithelial scrapings of the hamsters fed the ratio of 1:3.7. Mostly, pathways involved in lipid and energy metabolism were affected by the ratio of 1:3.7. PMID- 19217926 TI - Aspects of the neuroendocrine cerebellum: expression of secretogranin II, chromogranin A and chromogranin B in mouse cerebellar unipolar brush cells. AB - Morphologically distinct neuron classes can be subdivided in sublineages by differential chemical phenotypes that correlate with functional diversity. Here we show by immunocytochemistry that chromogranin A (CgA) chromogranin B (CgB) and secretogranin II (SgII), the principal granins situated in neuronal secretory granules and large dense-core vesicles, are widely but differentially expressed in cells of the mouse cerebellum and terminals of cerebellar afferents. While CgA and CgB were nearly panneuronal, SgII was more restricted in distribution. The cells most intensely immunoreactive for SgII were a class of small, excitatory interneurons enriched in the granular layer of the vestibulocerebellum, the unipolar brush cells (UBCs), although larger neurons likely to be a subset of the Golgi-Lugaro-globular cell population were also distinctly immunopositive; by contrast, Purkinje cells and granule cells were, at best, faintly stained and, stellate, basket cells were unstained. SgII was also present in subsets of mossy fibers, climbing fibers and varicose fibers. Neurons in the cerebellar nuclei and inferior olive were distinctly positive for the three granins. Double-labeling with subset-specific cell class markers indicated that, while both CgA and CgB were present in most UBCs, SgII immunoreactivity was present in the calretinin (CR)-expressing subset, but lacked in metabotropic glutamate receptor 1alpha (mGluR1alpha)-expressing UBCs. Thus, we have identified an additional cell class marker, SgII, which serves to study subtype properties in the UBC population. The abundance of SgII in only one of the two known subsets of UBCs is remarkable, as its expression in other neurons of the cortex was moderate or altogether lacking. The data suggest that the CR-positive UBCs represent a unique neuroendocrine component of the mammalian cerebellar cortex, presumably endowed with transynaptically regulated autocrine or paracrine action/s. Because of the well known organization of the cerebellar system, several of its neuron classes may represent valuable cellular models to analyze granin functions in situ, in acute slices and in dissociated cell and organotypic slice cultures. PMID- 19217927 TI - Generation, long-term persistence, and neuronal differentiation of cells with nuclear aberrations in the adult zebrafish brain. AB - Zebrafish, like other teleosts, continuously produce new cells in numerous regions of the adult brain. Immunolabeling employing antisera against phosphorylated histone-H3 and 5-bromo-2'-deoxyuridine revealed that approximately 6%-7% of such cells exhibited nuclear aberrations. These aberrations, presumably the result of mitotic segregation defects, included single and multiple laggards (both during metaphase and anaphase) and anaphase bridges. Cells with such aberrations persisted long-term and comprised, when examined 7.5 months after their generation, approximately 2.5% of the total population of adult-born cells. The drop in relative frequency of aberrations in the course of further development appears to be caused by elimination of cells with nuclear aberrations, presumably by apoptotic cell death. The cells with nuclear aberrations that persisted long-term were capable of neuronal differentiation, as demonstrated by combining anti-5-bromo-2'-deoxyuridine immunohistochemistry with immunostaining against the neuronal marker protein Hu or the enzyme tyrosine hydroxylase, a marker of catecholaminergic neurons. We hypothesize that the alterations in chromosome number and/or chromosome structure caused by nuclear aberrations do not necessarily result in loss of vital functions or in tumorigenesis. Instead, cells with such aberrations are able to undergo what appears to be normal development. PMID- 19217928 TI - Three sequential brain activations encode mental transformations of upright and inverted human bodies: a high resolution evoked potential study. AB - Human bodies provide a particularly rich source of visual information. Whereas most previous studies have focused on the neural mechanisms during the perception and recognition of human bodies, the aim of the present study was to investigate the time course and location of brain activation during mental imagery of human bodies. When participants were asked to imagine themselves in the position of a visually presented human body as seen from many different angles and at two orientations (upright or inverted), their reaction times were faster for upright as compared to inverted bodies and correlated differently with the tested angles. These behavioral effects were also reflected in brain activation patterns, but only during the time period from 220 to 490 ms after stimulus onset. Evoked potential mapping and electrical neuroimaging revealed three distinct and sequential steps of processing related to mental body transformation: (1) an early activation in temporo-occipital and temporo-parietal cortex (220-360 ms) that does not distinguish between upright and inverted bodies, but closely reflects the effort of mental transformation, followed (2) by an activation in temporo-occipital and medial parieto-occipital cortex (350-460 ms) that encodes mental transformation for upright bodies, and (3) a later activation in temporo occipital and prefrontal cortex (390-490 ms) that encodes mental transformation for inverted bodies. These data suggest that the mental transformation of human bodies is not a single process but a sequence of temporally distinct processing steps, where each step reflects a distinct aspect of the transformation process that consists of activations in a network of posterior brain areas including extrastriate cortex, temporo-parietal cortex, and medial parieto-occipital cortex, as well as an anterior brain region in prefrontal cortex. PMID- 19217929 TI - Brief constant light accelerates serotonergic re-entrainment to large shifts of the daily light/dark cycle. AB - Brief ( approximately 2 day) constant light exposure (LL(b)) in hamsters dramatically enhances circadian phase-resetting induced by the 5-HT receptor agonist, (+/-)-2-dipropyl-amino-8-hydroxyl-1,2,3,4-tetrahydronapthalene (8-OH DPAT) and other nonphotic stimuli. The present study was undertaken to determine if LL(b) can also amplify phase-resetting responses to endogenous 5-HT and accelerate re-entrainment to large-magnitude advance and delay shifts of the light/dark (LD) cycle. First, central serotonergic activity was increased by i.p. injection of L-tryptophan+/-the 5-HT reuptake inhibitor fluoxetine. Hamsters under LD or exposed to LL(b) received vehicle or drugs during the early morning, and phase-shifts of the locomotor activity rhythm were measured after release to constant darkness. Neither drug phase-shifted animals not exposed to LL(b) (P>0.5 vs. vehicle); however in animals receiving LL(b,)L-tryptophan with and without fluoxetine produced large phase-advance shifts (means=2.5+/-0.4 h and 2.6+/-0.2 h, respectively; both P<0.035 vs. vehicle). Next, the effects of LL(b) combined with 8-OH-DPAT or L-tryptophan+fluoxetine on serotonergic re-entrainment to 10 h phase-advance and phase-delay shifts of the LD cycle were assessed. In groups not exposed to LL(b), vehicle controls re-entrained slowly to the advance and delay shifts (means=16+/-1 and 24+/-4 days, respectively), but those treated with 8-OH DPAT re-entrained faster (means=11+/-2 and 9+/-2 days, respectively; both P<0.05 vs. vehicle). In groups exposed to LL(b), vehicle controls re-entrained slowly to the advance and delay shifts (means=15+/-2 and 25+/-3 days, respectively); however those receiving 8-OH-DPAT rapidly re-entrained to the delay and advance shifts, with the majority (75%) requiring only 1-2 days (means=2+/-1 and 4+/-2 days, respectively; both P<0.05 vs. vehicle). Animals exposed to LL(b) and treated with L-tryptophan+fluoxetine also exhibited accelerated re-entrainment to a 10 h advance shift (mean=5+/-2 days; P<0.05 vs. vehicle). Thus through enhancing serotonergic phase-resetting, LL(b) facilitates rapid re-entrainment to large shifts of the LD cycle which offers a potential approach for treating circadian-related desynchronies. PMID- 19217930 TI - GABAA receptors in the mediodorsal thalamus play a crucial role in rat shell specific acetylcholine-mediated, but not dopamine-mediated, turning behaviour. AB - The role of GABA(A) receptors in the mediodorsal thalamus (mdT) in turning behaviour of rats was studied. Neither the GABA(A) receptor agonist muscimol (50 ng) nor the antagonist bicuculline (200 ng) unilaterally injected into the mdT elicited any behavioural change. Unilateral injection of the acetylcholine receptor agonist (carbachol, 5 microg) into the nucleus accumbens shell has been found to elicit contraversive circling while unilateral injection of a mixture of dopamine D(1) ((+/-)-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine-7,8-diol [SKF 38393], 5 microg) and D(2) (quinpirole, 10 microg) receptor agonists into the same site is known to elicit contraversive pivoting. The contraversive circling induced by unilateral injection of carbachol (5 microg) into the nucleus accumbens shell was dose-dependently inhibited by muscimol (25 and 50 ng) injected into the mdT. This inhibitory effect of muscimol (50 ng) was antagonised by co-administration of bicuculline (200 ng), which alone did not modify the contraversive circling induced by carbachol (5 microg). The contraversive pivoting induced by unilateral injection of a mixture of SKF 38393 (5 microg) and quinpirole (10 microg) into the nucleus accumbens shell was inhibited by muscimol (25 and 50 ng) injected into the mdT, whereas bicuculline (200 ng) injected into the mdT did not significantly modify the pivoting. The inhibitory effect of muscimol (50 ng) on the pivoting induced by a mixture of SKF 38393 (5 microg) and quinpirole (10 microg) was not dose-dependent and not antagonised by bicuculline (200 ng). The present study suggests that GABA(A) receptors in the mdT play a limited role in spontaneously occurring locomotor activity. Secondly, this study demonstrates that GABA(A) receptors in the mdT transmit accumbens-dependent cholinergic circling, but not accumbens-dependent dopaminergic pivoting, to other brain structures. Finally, the present study shows that muscimol-sensitive, non GABA(A) receptors in the mdT influence the accumbens-dependent dopaminergic pivoting. To what extent GABA(B) receptors in the mdT mediate the muscimol induced effects upon the dopaminergic pivoting behaviour requires additional research. PMID- 19217933 TI - Novel, continuous visual motion induces c-fos expression in the avian optokinetic nuclei and optic tectum. AB - We studied the stimulus characteristics necessary for the expression of c-fos protein in optokinetic system neurons using immunocytochemistry. Using whole field visual motion as a stimulus, we found substantial c-fos expression in the optic tectum (TeO), the nucleus of the basal optic root (nBOR) and the pretectal nucleus lentiformis mesencephali (LM); in all cases immunostaining was seen only on the side contralateral to the eye viewing whole-field unidirectional motion; the side of the brain contralateral to the eye wearing a diffuser showed no staining. In the nBOR and the LM, different regions showed a remarkable specificity of c-fos expression depending on the direction of visual motion stimulation. Neurons were stained primarily in regions known from previous electrophysiological recordings to be maximally responsive to that direction of motion; little staining was seen after motion orthogonal to the preferred motion direction. Novel, continuous visual motion stimuli, lasting more than 30 min, was required for maximal c-fos expression, suggesting that brief periods of unidirectional optic flow, as would be experienced during normal life, do not stimulate the expression of c-fos. The largest number of neurons was labeled when birds raised from hatching with one eye covered by a diffuser were exposed to full-field visual motion immediately after the diffuser was switched from one eye to the other, so that only the previously naive eye was visually stimulated. We conclude that the expression of c-fos in the optokinetic nuclei is linked to near peak firing rates on the one hand, and the novelty and duration of the visual signals, on the other, supporting the assumption that this expression is mainly related to stimulus contexts leading to neuronal plastic changes. PMID- 19217932 TI - Inositol 1,4,5-triphosphate receptors and NAD(P)H mediate Ca2+ signaling required for hypoxic preconditioning of hippocampal neurons. AB - Exposure of neurons to a non-lethal hypoxic stress greatly reduces cell death during subsequent severe ischemia (hypoxic preconditioning, HPC). In organotypic cultures of rat hippocampus, we demonstrate that HPC requires inositol triphosphate (IP3) receptor-dependent Ca2+ release from the endoplasmic reticulum (ER) triggered by increased cytosolic NAD(P)H. Ca2+ chelation with intracellular BAPTA, ER Ca2+ store depletion with thapsigargin, IP3 receptor block with xestospongin, and RNA interference against subtype 1 of the IP3 receptor all blunted the moderate increases in [Ca2+](i) (50-100 nM) required for tolerance induction. Increases in [Ca2+](i) during HPC and neuroprotection following HPC were not prevented with NMDA receptor block or by removing Ca2+ from the bathing medium. Increased NAD(P)H fluorescence in CA1 neurons during hypoxia and demonstration that NADH manipulation increases [Ca2+](i) in an IP3R-dependent manner revealed a primary role of cellular redox state in liberation of Ca2+ from the ER. Blockade of IP3Rs and intracellular Ca2+ chelation prevented phosphorylation of known HPC signaling targets, including MAPK p42/44 (ERK), protein kinase B (Akt) and CREB. We conclude that the endoplasmic reticulum, acting via redox/NADH-dependent intracellular Ca2+ store release, is an important mediator of the neuroprotective response to hypoxic stress. PMID- 19217931 TI - Age-related changes in glycine receptor subunit composition and binding in dorsal cochlear nucleus. AB - Age-related hearing loss, presbycusis, can be thought of, in part, as a slow progressive peripheral deafferentation. Previous studies suggest that certain deficits seen in presbycusis may partially result from functional loss of the inhibitory neurotransmitter glycine in dorsal cochlear nucleus (DCN). The present study assessed age-related behavioral gap detection changes and neurochemical changes of postsynaptic glycine receptor (GlyRs) subunits and their anchoring protein gephyrin in fusiform cells of young (7-11 months) and aged (28-33 months) Fischer brown Norway (FBN) rats. Aged rats showed significantly (20-30 dB) elevated auditory brainstem-evoked response thresholds across all tested frequencies and worse gap detection ability compared to young FBN rats. In situ hybridization and quantitative immunocytochemistry were used to measure GlyR subunit message and protein levels. There were significant age-related increases in the alpha(1) subunit message with significant age-related decreases in alpha(1) subunit protein. Gephyrin message and protein showed significant increases in aged DCN fusiform cells. The pharmacologic consequences of these age related subunit changes were assessed using [3H] strychnine binding. In support of the age-related decrease of alpha(1) subunit protein levels in DCN, there was a significant age-related decrease in the total number of GlyR binding sites with no significant change in affinity. These age-related changes may reflect an effort to reestablish a homeostatic balance between excitation and inhibition impacting on DCN fusiform cells by downregulation of inhibitory function in the face of an age-related loss of peripheral input. Age-related decrease in presynaptic glycine release results in altered subunit composition and this may correlate with loss of temporal coding of the aged fusiform cell in DCN. The previously reported role for gephyrin in retrograde intracellular receptor subunit trafficking could contribute to the alpha(1) decrease in the face of increased message. PMID- 19217934 TI - Energy-restricted pair-feeding normalizes low levels of brain-derived neurotrophic factor/tyrosine kinase B mRNA expression in the hippocampus, but not ventromedial hypothalamic nucleus, in diet-induced obese mice. AB - Brain-derived neurotrophic factor (BDNF) and its receptor tyrosine kinase B (TrkB) are closely associated with the regulation of energy homeostasis, but their roles in diet-induced obesity have not been explored. Using dietary interventions, this study examined regional changes of BDNF and TrkB mRNA expression in different brain regions of diet-induced obese (DIO) and resistant (DR) mice in response to high-fat (HF), energy-restricted pair-feeding and low fat (LF) diets. Using in situ hybridization, DIO mice had significantly decreased levels of BDNF mRNA expression (-32% to -37%) and TrkB (-21% to -23%) in the hippocampus compared to DR mice on an HF diet, but not on energy-restricted pair feeding and LF diets. In the ventromedial hypothalamic nucleus (VMH), BDNF expression was decreased in DIO mice on HF (-23%) and energy-restricted pair feeding (-21%) diets. Furthermore, the VMH BDNF expression was negatively correlated with blood glucose but positively correlated with plasma adiponectin. These findings suggest that decreased hippocampal BDNF and TrkB expression plays an important role in high-fat diet induced obesity. A lower baseline BDNF mRNA expression in the VMH of DIO mice after normalization of body weight may indicate their intrinsic nature or an elevated body weight set point to drive body weight gain. PMID- 19217935 TI - Displacements of the organ of Corti by gel injections into the cochlear apex. AB - In order to transduce sounds efficiently, the stereocilia of hair cells in the organ of Corti must be positioned optimally. Mechanical displacements, such as pressure differentials across the organ caused by endolymphatic hydrops, may impair sensitivity. Studying this phenomenon has been limited by the technical difficulty of inducing sustained displacements of stereocilia in vivo. We have found that small injections (0.5-2 microL) of Healon gel into the cochlear apex of guinea pigs produced sustained changes of endocochlear potential (EP), summating potential (SP) and transducer operating point (OP) in a manner consistent with a mechanically-induced position change of the organ of Corti in the basal turn. Induced changes immediately recovered when injection ceased. In addition, effects of low-frequency bias tones on EP, SP and OP were enhanced during the injection of gel and remained hypersensitive after injection ceased. This is thought to result from the viscous gel mechanically limiting pressure shunting through the helicotrema. Cochlear microphonics measured as frequency was varied showed enhancement below 100 Hz but most notably in the sub-auditory range. Sensitivity to low-frequency biasing was also enhanced in animals with surgically-induced endolymphatic hydrops, suggesting that obstruction of the perilymphatic space by hydrops could contribute to the pathophysiology of this condition. PMID- 19217936 TI - Circulating levels of soluble intercellular adhesion molecule-1 (sICAM-1) independently predict depressive symptom severity after 12 months in heart failure patients. AB - OBJECTIVE: To determine whether inflammatory markers prospectively predict depressive symptom severity 12 months later in heart failure (HF) patients. METHODS: In 30 HF patients we assessed depressive symptom severity by the Beck depression inventory (BDI) at baseline as well as 12 months later. We measured circulating levels of the soluble intercellular adhesion molecule (sICAM)-1, the cytokine interleukin (IL)-6 and the acute phase protein C-reactive protein (CRP) at baseline assessment. RESULTS: sICAM-1 (r=.38, p=.045) but not CRP or IL-6 correlated with BDI scores 12 months later. Hierarchical linear regression analysis revealed that independent of baseline BDI assessment, cardiovascular risk factors, indicators of HF disease severity, and medication intake, sICAM-1 significantly predicted BDI scores 12 months later. sICAM-1 independently explained between 7% (beta=.26, p=.040) and 10% (beta=.35, p=.045) of the total variance in BDI scores 12 months later. CONCLUSION: The findings from this exploratory analysis suggest that the adhesion molecule sICAM-1 is an independent predictor of depressive symptoms 12 months later in HF patients. Our prospective findings support the suggested role for inflammation in increasing future depressive symptom severity and extend this linkage for the first time to HF. PMID- 19217937 TI - Fetal origins of mental health: evidence and mechanisms. AB - The concept of fetal programming states that changes in the fetal environment during sensitive periods of organ development may cause long-lasting changes in the structure and functioning of these organs later in life and influence the risk for chronic diseases such as coronary heart disease and type 2 diabetes. Fetal growth is a summary marker of the fetal environment and is reflected by relatively easy-to-obtain measures of size at birth such as birth weight. In the last two decades, a body of evidence emerged linking fetal growth with behavioural and mental health outcomes later in life. Cognitive functioning and behavioural problems in childhood, in particular inattention/hyperactivity, have been shown to be inversely related to fetal growth. Although results are mixed, risk for personality disorders and schizophrenia seems to be linked with fetal growth and adversity, while the evidence for mood disorders is weak. Vulnerability for psychopathology may also be influenced by prenatal adversity. There is evidence for associations of fetal growth with temperament in childhood as well as stress reactivity and distress. The associations of fetal growth with mental health later in life are potentially caused by specific prenatal factors such as maternal smoking, alcohol, toxins/drugs, nutrition, psychosocial stress and infection during pregnancy. The mechanisms likely involve changes in neurodevelopment and in the set point of neuroendocrine systems, and there is evidence that prenatal adversity interacts with genetic and postnatal environmental factors. Future studies should examine the effects of specific prenatal factors and attempt to disentangle genetic and prenatal environmental effects. PMID- 19217938 TI - Psychological stress suppresses innate IFN-gamma production via glucocorticoid receptor activation: reversal by the anxiolytic chlordiazepoxide. AB - Studies in humans and in animals indicate that psychological stress can modulate immune responses. Here we demonstrate that exposure to psychological stress (restraint stress) suppresses innate interferon (IFN)-gamma production in mice following an in vivo lipopolysaccharide (LPS) challenge. IFN-gamma signaling was also impaired by stress, as indicated by reduced STAT1 phosphorylation and reduced expression of the IFN-gamma-inducible genes, inducible nitric oxide synthase (iNOS) and IFN-gamma-inducible protein 10 (IP-10/CXCL10). Furthermore, restraint stress suppressed production of the IFN-gamma inducing cytokine interleukin (IL)-12 and increased production of the anti-inflammatory cytokine IL 10, which can inhibit both IL-12 and IFN-gamma production. However, using IL-10 knockout mice, we demonstrate that IL-10 does not mediate the suppressive effect of restraint stress on innate IFN-gamma production. Restraint stress increased corticosterone concentrations in serum and spleen, and consistent with a role for glucocorticoids in the immunosuppressive actions of stress, pre-treatment with the glucocorticoid receptor antagonist mifepristone completely blocked the stress related suppression of innate IFN-gamma production. Addition of exogenous IL-12 to LPS-stimulated spleen cells reversed the suppressive effect of both restraint stress and corticosterone on IFN-gamma production. These data suggest that reduced IL-12 production is a key event in stress-induced suppression of innate IFN-gamma production. Finally, we demonstrate that pre-treatment with the anxiolytic drug chlordiazepoxide prevents the suppressive effect of stress on innate IFN-gamma production, and also attenuates the stress-induced increase in circulating corticosterone concentrations. PMID- 19217939 TI - Psychologically adverse work conditions are associated with CD8+ T cell differentiation indicative of immunesenescence. AB - Numerous studies have demonstrated associations between psychosocial stress and indices of poor health, and much research is now dedicated to identifying the responsible biological mechanisms. The current study examined the hypothesis that stress may impact health by promoting immunesenescence. Participants were 537 factory workers (89% male; mean age 44; range 18-65years). Blood was analyzed for two components of the aging 'immune risk phenotype': the number and proportion of late-differentiated (CD27-CD28-) CD8 T cells (CTLs) and CD4:CD8 ratio. Psychological assessment focussed on work-related stressors which have previously been found to predict morbidity and mortality. This assessment included measures of work load, effort-reward imbalance, and social support at work. High levels of job stress (low reward, high effort-reward imbalance) and low social support at work were associated with a significantly lower CD4:CD8 ratio. Also, the number of CD27-CD28- CTLs was 30% to 50% higher in employees classified in the highest tertile of each stress parameter as compared to employees in the corresponding lowest tertile (p<.01). These associations withstood adjustment for a wide range of demographic, life style, medical, and socio-economic indicators. The associations between CTL phenotype and low social support became stronger with increasing age. These results suggest that psychosocial stress may contribute to immunological aging. Prospective studies should address the long-term consequences of these associations for healthy aging. PMID- 19217940 TI - A mechanism-based complementary screening approach for the amelioration and reversal of neurobehavioral teratogenicity. AB - The identification of mechanisms and outcomes for neurobehavioral teratogenesis is critical to our ability to develop therapies to ameliorate or reverse the deleterious effects of exposure to developmental neurotoxicants. We established mechanistically-based complementary models for the study of cholinergic systems in the mouse and the chick, using both environmental neurotoxicants (chlorpyrifos, perfluoroalkyls) and drugs of abuse (heroin, nicotine, PCP). Behavioral evaluations were made using the Morris maze in the mouse, evaluating visuospatial memory related to hippocampal cholinergic systems, and imprinting in the chick, examining behavior dependent on cholinergic innervation of the IMHV. In both models we demonstrated the dependence of neurobehavioral deficits on impairment of cholinergic receptor-induced expression, and translocation of specific PKC isoforms. Understanding this mechanism, we were able to reverse both the synaptic and behavioral deficits with administration of neural progenitors. We discuss the prospects for clinical application of neural progenitor therapy, emphasizing protocols for reducing or eliminating immunologic rejection, as well as minimizing invasiveness of procedures through development of intravenous administration protocols. PMID- 19217941 TI - Functional expression and sub-cellular localization of the early aflatoxin pathway enzyme Nor-1 in Aspergillus parasiticus. AB - Aflatoxin biosynthesis in Aspergillus parasiticus requires at least 17 enzyme activities (from acetate). Although the activities of most aflatoxin biosynthetic enzymes have been established, the mechanisms that govern transport and sub cellular localization of these enzymes are not clear. We developed plasmid constructs that express Nor-1 fused to a green fluorescent protein reporter (EGFP) to monitor transport and localization of this early pathway enzyme in real time in Aspergillus parasiticus. Plasmids expressing EGFP fused to Nor-1 were introduced into A. parasiticus B62 (carries non-functional Nor-1). Transformants were screened for increased aflatoxin accumulation (restored Nor-1 activity) on coconut agar medium and for EGFP expression using fluorescence microscopy. Increased aflatoxin accumulation was confirmed by TLC and ELISA. Nor-1 fused to EGFP at either the N- or C- terminus functionally complemented non-functional Nor 1 in B62 and increased aflatoxin synthesis to wild-type (N-terminus) or lower levels (C-terminus). We detected full-length Nor-1 fusion proteins in transformants with increased aflatoxin accumulation (Western blot) and determined that the expression plasmid integrated at the nor-1 locus in these cells (Southern blot). Confocal laser scanning microscopy (CLSM) demonstrated that Nor 1 fusion proteins localized in the cytoplasm and vacuoles of fungal hyphae grown on aflatoxin-inducing solid media for 48h; control EGFP (no Nor-1) did not localize to vacuoles until 72h. The highest rate of aflatoxin synthesis coincided with the highest rate of transport of Nor-1 fusion proteins to the vacuole strongly suggesting that Nor-1 is synthesized in the cytoplasm and transported to the vacuole to carry out an early step in aflatoxin synthesis. PMID- 19217942 TI - Soluble expression and one-step purification of a neurotoxin Huwentoxin-I in Escherichia coli. AB - Huwentoxin-I (HWTX-I) is a small 33-amino acid neurotoxin from the venom of the Chinese bird spider Ornithoctonus huwena. HWTX-I selectively blocks N-type voltage-sensitive calcium channels (N-VSCCs) and has great potential for clinical application as a novel analgesic without inducing drug tolerance. However, there are still many unsolved issues for this peptide, such as its clinical efficacy in analgesia, anesthesia, and even its potential role in drug rehabilitation. Therefore, large amounts of active recombinant HWTX-I are urgently needed. In this report, we describe a novel and efficient way to produce large amounts of the valuable form in Escherichia coli. HWTX-I was expressed in soluble form as an N-terminal intein fusion product. After affinity purification, a pH shift-induced self-cleavage of the intein released HWTX-I, resulting in a single-column purification of the target protein. The whole-cell patch clamp assay showed that purified HWTX-I has activity similar to another commercialized N-VSCC blocker omega-conotoxin MVIIA. Production of HWTX-I by this method has the major advantages of high efficiency and low cost. PMID- 19217943 TI - Transient amorphous calcium phosphate in forming enamel. AB - Enamel, the hardest tissue in the body, begins as a three-dimensional network of nanometer size mineral particles, suspended in a protein gel. This mineral network serves as a template for mature enamel formation. To further understand the mechanisms of enamel formation we characterized the forming enamel mineral at an early secretory stage using X-ray absorption near-edge structure (XANES) spectromicroscopy, transmission electron microscopy (TEM), FTIR microspectroscopy and polarized light microscopy. We show that the newly formed enamel mineral is amorphous calcium phosphate (ACP), which eventually transforms into apatitic crystals. Interestingly, the size, shape and spatial organization of these amorphous mineral particles and older crystals are essentially the same, indicating that the mineral morphology and organization in enamel is determined prior to its crystallization. Mineralization via transient amorphous phases has been previously reported in chiton teeth, mollusk shells, echinoderm spicules and spines, and recent reports strongly suggest the presence of transient amorphous mineral in forming vertebrate bones. The present finding of transient ACP in murine tooth enamel suggests that this strategy might be universal. PMID- 19217944 TI - Candida albicans triggers proliferation and differentiation of hematopoietic stem and progenitor cells by a MyD88-dependent signaling. AB - As TLRs are expressed by hematopoietic stem and progenitor cells, these receptors may play a role in hematopoiesis in response to pathogens during infection. We showed here that inactivated yeasts and hyphae of Candida albicans induce in vitro the proliferation of purified murine hematopoietic stem and progenitor cells (Lin(-)c-Kit(+) Sca-1(+)) as well as their differentiation to lineage positive cells, through a MyD88-dependent pathway. These results indicate that TLR-mediated recognition of C. albicans by hematopoietic stem and progenitor cells may augment the host capability for rapidly replenishing the innate immune system during candidiasis. PMID- 19217945 TI - Physiological effects of tricyclazole on zebrafish (Danio rerio) and post exposure recovery. AB - Short-term effects of tricyclazole on male zebrafish (Danio rerio) physiology were examined joint to the degree of recovery after exposure. Fish were exposed to 142 microg/L (1/100 LC(50)-96 h) of tricyclazole for 7 (Exp.1) and 14 days (Exp.2) and then allowed to recover for 7 or 14 more days, respectively. Whole body triglycerides, cholesterol, glucose, lactate and total proteins were measured as well as the aspartate aminotransferase (AAT), alanine aminotransferase (AlAT), alkaline phosphatase (AP) and lactate dehydrogenase (LDH) activities as biomarkers of intermediary metabolism; gamma-glutamyl transpeptidase (gammaGT) as biomarker of oxidative detoxification processes and vitellogenin (Vtg) concentration as endpoint for endocrine disruptor effect were also determined. Corpulence factor (k) was calculated. Fungicide exposure in zebrafish resulted in an increased of triglycerides, cholesterol, glucose and lactate levels, however the total protein content did not change. LDH, AlAT and AAT enhanced while AP activity decreased. Corpulence factor (k) also decreased. At the end of the recovery periods cholesterol and glucose levels recovered whereas triglycerides and lactate continued to elevate. Induced disorders on the selected enzymes remained and did not recover at the end of experiments. Fish exhibited significant increases of Vtg during the overall experimental times as a consequence of the fungicide exposure. These findings are of importance in the assessment of the potential risk of new fungicides as tricyclazole on aquatic ecosystems. PMID- 19217947 TI - Specificity of islet cell autoantibodies and coexistence with other organ specific autoantibodies in type 1 diabetes mellitus. AB - Type 1 diabetes mellitus (T1DM) has been shown to be a disease characterized by immune-mediated destruction of the insulin-producing islet beta-cells (beta cells) in the pancreas. Intensive studies, in both patients and animal models are trying to elucidate the specific antigenic targets that are responsible for islet cell autoimmunity. So far, the most important molecules that have been recognized are the native insulin, the 65-kDa form of glutamic acid decarboxylase (GAD(65)) and the insulinoma-antigen 2 (IA-2). Identification of those specific autoantibodies that are involved in the primary immunological events of the autoimmune disease process will allow the development of novel diagnostic procedures for early detection and initiation of potential therapy prior to irreversible loss of beta-cells. Within the framework of polyglandular disorders, T1DM may coexist with other organ specific autoimmune diseases such as autoimmune thyroid disease (ATD), autoimmune gastritis (AG), celiac disease (CD) and Addison's disease (AD), which are associated with the production of organ specific autoantibodies. So, as a subset of patients with those autoantibodies will develop clinical disease, screening T1DM patients could prognosticate morbidity relative to unrecognised clinical entities. The close follow-up of patients with organ-specific autoantibodies could lead to seasonable identification of those requiring therapy. PMID- 19217948 TI - Proteomic approaches to study structure, functions and toxicity of legume seeds lectins. Perspectives for the assessment of food quality and safety. AB - Lectins are a structurally diverse class of (glyco)proteins which bind mono- and oligosaccharides with high specificity and in a reversible way. For many years, the unique sugar binding properties of plant lectins have been exploited for the development of biochemical tools for glycoprotein isolation and characterisation, and the use of lectins as a glycoprofiling tool has became much more sophisticated with the advent of lectin microarrays, in which a panel of lectins are immobilized on a single chip for glycomic analysis. Among the numerous lectins studied so far, those from legumes represent the largest family. They can be present at relatively high amounts depending on genetic as well as environmental factors, and are accumulated especially in the seeds. For this reason, some lectins as the phytohemagglutinin from the common bean Phaseolus vulgaris constitute a possible risk, since consumption of raw or incorrectly processed beans has been shown to cause outbreaks of gastroenteritis, nausea and diarrhoea. On the other hand, for these anti-nutritional properties, bean extracts enriched in lectins or in lectin-related amylase inhibitors are also finding a growing use as active ingredients of "weight-blockers" in dietetic preparations for obesity treatment. Current methods to determine the lectin levels in foods are based on immunoenzymatic or toxicity tests, which are largely aspecific. Very recently, the availability of proteomic methodologies has allowed to start development and validation of sensitive and specific assays for detecting trace amounts of harmful lectins in either raw or processed foods. In this review, the main aspects of current and perspective applications of mass spectrometry and proteomic technologies to the structural characterisation of legumes are presented, with focus on issues related to detection, identification, and quantification of phytohemagglutinins relevant for their biochemical, immunological and toxicological aspects. PMID- 19217946 TI - Angiogenesis and its targeting in rheumatoid arthritis. AB - Angiogenesis, the development of new capillaries, is involved in leukocyte ingress into the synovium during the development and progression of rheumatoid arthritis. Several soluble and cell surface-bound mediators including growth factors, cytokines, chemokines, proteolytic matrix-degrading enzymes, cell adhesion molecules and others may promote synovial neovascularization. On the other hand, endogenous angiostatic factors, such as angiostatin, endostatin, interleukin-4 (IL-4), IL-13, interferons and some angiostatic chemokines are also produced within the rheumatoid synovium, however, their effects are insufficient to control synovial angiogenesis and inflammation. Several specific and non specific strategies have been developed to block the action of angiogenic mediators. The first line of angiostatic agents include vascular endothelial growth factor (VEGF), angiopoietin, alpha(V)beta(3) integrin antagonist, as well as non-specific angiogenesis inhibitors including traditional disease-modifying agents (DMARDs), anti-tumor necrosis factor biologics, angiostatin, endostatin, fumagillin analogues or thalidomide. Potentially any angiostatic compound could be introduced to studies using animal models of arthritis or even to human rheumatoid arthritis trials. PMID- 19217950 TI - Sucking pump activity in feeding behaviour regulation in carpenter ants. AB - Modulation of liquid feeding-rate would allow insects to ingest more food in the same time when this was required. Ants can vary nectar intake rate by increasing sucking pump frequency according to colony requirements. We analysed electrical signals generated by sucking pump activity of ants during drinking solutions of different sucrose concentrations and under different carbohydrate-deprivation levels. Our aim was to define parameters that characterize the recordings and analyse their relationship with feeding behaviour. Signals showed that the initial and final frequencies of sucking pump activity, as well as the difference between them were higher in sugar-deprived ants. However, these parameters were not influenced by sucrose solution concentration, which affected the number of pump contractions and the volume per contraction. Unexpectedly, we found two different responses in feeding behaviour of starved and non-starved ants depending on concentration. Starved ants drank dilute solutions for the same length of time as non-starved ants but ingested higher volumes. While drinking the concentrated solutions, starved ants drank the same volume, but did so in a shorter time than the non-starved ones. Despite these differences, for each analysed concentration the total number of pump contractions remained constant independently of sugar-deprivation level. These results are discussed in the frame of feeding regulation and decision making in ant foraging behaviour. PMID- 19217949 TI - The homeodomain of PAX6 is essential for PAX6-dependent activation of the rat glucagon gene promoter: evidence for a PH0-like binding that induces an active conformation. AB - The transcription factor PAX6 plays an important role in transcriptional regulation of the peptide hormone glucagon from pancreatic alpha-cells. PAX6 contains two DNA binding domains, the paired domain (PD) and the homeodomain (HD). While the interaction of the PD with the PAX6 responsive elements G1 and G3 in the rat glucagon gene promoter is well understood, the role of the PAX6 HD for PAX6 binding and function on G1 and G3 remains unclear. In EMSA studies the PAX6 HD was found to be mandatory for PAX6 binding to G1 but not to G3. Transient transfections with luciferase reporter gene constructs revealed the HD to be critical for proper function of PAX6 on both, G1 and G3. Transfection data with variant promoter constructs and limited proteolysis assays demonstrated that the DNA sequence located 5' to the PD binding site plays an important role for PAX6 function and its conformation on the elements G1 and G3. Taken together, our data indicate a PH0-like binding of PAX6 to the glucagon promoter elements G1 and G3 where the HD binding site is abutted directly to the PD binding motif. The data suggest that the PH0-like binding induces a transcriptionally active conformation of PAX6. PMID- 19217951 TI - The logic behind target intraocular pressure. PMID- 19217952 TI - Macula hole surgery: simple or complex? PMID- 19217953 TI - Controversy over "contradiction": Should randomized trials always trump observational studies? PMID- 19217954 TI - The role of statistics in ophthalmology. PMID- 19217955 TI - Descriptive statistics in ophthalmic research. PMID- 19217956 TI - Complex palpebral odontogenic choristoma: A reappraisal of the origin of teeth bearing periocular lesions. AB - PURPOSE: To describe the findings observed in a congenital ectopic tooth of the lower inner eyelid and explain its origin. DESIGN: Retrospective and interventional clinicopathologic study. METHODS: Histopathologic evaluation of sequentially excised specimens over 4 years and radiographic study of any associated dental abnormalities, compared with critical reevaluation of similar reported cases in the literature. RESULTS: The ectopic eyelid tooth clinically created a dome-shaped mass and microscopically displayed advanced cytodifferentiation but incomplete morphogenesis with clearly identifiable ameloblasts, enamel, odontoblasts, and dentin, surrounded by an intact dental sac. Swollen oral mucosal epithelium with associated sebaceous Fordyce nodules, residual ciliated odontogenic epithelium, embryonic epithelial rests, and accessory mucinous and sero-mucinous accessory salivary glands were discovered in place of normal conjunctiva. The tarsus with meibomian glands was absent but islands of cartilage were present. CONCLUSIONS: Because of the variety of the ectopic tissues that were exhibited, a causative displacement of oral embryonic epithelium with extensive differentiating potentialities is posited. Two previously reported palpebral dental choristomas also had identical locations in the inferomedial eyelid. This is the site where the phakomatous choristoma may be encountered, but this unencapsulated mass creates an eyelid thickening without a globoid configuration and infiltrates the inferior orbit. After a careful re appraisal of published reports of orbital cysts bearing a tooth that were formerly diagnosed as "teratomas," we have concluded that they developed from similar embryonic oral (buccal) epithelial displacements rather than from aberrant germ cells, based on the cysts' linings and the associated accessory salivary glands in their walls. This view is further supported by the absence of heterologous teratomatous elements of endodermal or neuroectodermal derivation. A clinical and surgical approach to management and a new diagnostic schema for odontogenic choristomas are proposed. PMID- 19217957 TI - The need for standardization of antiretinal antibody detection and measurement. PMID- 19217959 TI - Not for your eyes: information concealed through publication bias. PMID- 19217961 TI - Optical coherence tomography findings in shaken baby syndrome: Forbes editorial. PMID- 19217964 TI - Regarding: ocular coherence tomography in shaken baby syndrome. PMID- 19217965 TI - Debunking myths in physician-industry conflicts of interest. PMID- 19217967 TI - The Neuroscience Peer Review Consortium. PMID- 19217968 TI - Resources pressures and small island states. PMID- 19217969 TI - A new agenda for children affected by HIV/AIDS. PMID- 19217970 TI - Data protection in a digital age. PMID- 19217971 TI - The long and short of dementia care in England. PMID- 19217972 TI - Shaping future health: Berlin Evolution of Medicine Summit. PMID- 19217973 TI - Prophylaxis of catheter-related thrombosis in cancer patients. PMID- 19217974 TI - Immobilisation for acute severe ankle sprain. PMID- 19217975 TI - Italian G8 Summit: a critical juncture for global health. PMID- 19217976 TI - 10 years of success in addressing lymphatic filariasis. PMID- 19217977 TI - Ticking the right boxes. PMID- 19217978 TI - Sidney Wolfe. PMID- 19217979 TI - Gefitinib plus docetaxel in non-small-cell lung cancer. PMID- 19217980 TI - Gefitinib plus docetaxel in non-small-cell lung cancer. PMID- 19217981 TI - Gefitinib plus docetaxel in non-small-cell lung cancer. PMID- 19217983 TI - Did he really bring the war home with him? PMID- 19217985 TI - Women and the global AIDS epidemic. PMID- 19217986 TI - Response to "Rosiglitazone no longer recommended". PMID- 19217987 TI - Combination HIV prevention must be based on evidence. PMID- 19217988 TI - Availability of drugs for formidable communicable diseases. PMID- 19217989 TI - Prevention of baby-walker-related injury. PMID- 19217990 TI - A medical maxim reinstated. PMID- 19217991 TI - Warfarin thromboprophylaxis in cancer patients with central venous catheters (WARP): an open-label randomised trial. AB - BACKGROUND: The role and dose of anticoagulants in thromboprophylaxis for patients with cancer receiving chemotherapy through central venous catheters (CVCs) is controversial. We therefore assessed whether warfarin reduces catheter related thrombosis compared with no warfarin and whether the dose of warfarin determines the thromboprophylactic effect. METHODS: In 68 clinical centres in the UK, we randomly assigned 1590 patients aged at least 16 years with cancer who were receiving chemotherapy through CVCs to no warfarin, fixed-dose warfarin 1 mg per day, or dose-adjusted warfarin per day to maintain an international normalised ratio between 1.5 and 2.0. Clinicians who were certain of the benefit of warfarin randomly assigned patients to fixed-dose or dose-adjusted warfarin groups. The primary outcome was the rate of radiologically proven, symptomatic catheter-related thrombosis. Analysis was by intention to treat. This trial is registered as an International Standard Randomised Controlled Trial, number ISRCTN 50312145. FINDINGS: Compared with no warfarin (n=404), warfarin (n=408; 324 [79%] on fixed-dose and 84 [21%] on dose-adjusted) did not reduce the rate of catheter-related thromboses (24 [6%] vs 24 [6%]; relative risk 0.99, 95% CI 0.57 1.72, p=0.98). However, compared with fixed-dose warfarin (n=471), dose-adjusted warfarin (n=473) was superior in the prevention of catheter-related thromboses (13 [3%] vs 34 [7%]; 0.38, 0.20-0.71, p=0.002). Major bleeding events were rare; an excess was noted with warfarin compared with no warfarin (7 vs 1, p=0.07) and with dose-adjusted warfarin compared with fixed-dose warfarin (16 vs 7, p=0.09). A combined endpoint of thromboses and major bleeding showed no difference between comparisons. We did not note a survival benefit in either comparison. INTERPRETATION: The findings show that prophylactic warfarin compared with no warfarin is not associated with a reduction in symptomatic catheter-related or other thromboses in patients with cancer and therefore we should consider newer treatments. FUNDING: Medical Research Council and Cancer Research UK. PMID- 19217993 TI - Hepatitis B virus infection. AB - Since the introduction of the hepatitis B vaccine and other preventive measures, the worldwide prevalence of hepatitis B infection has fallen. However, chronic infection remains a challenging global health problem, with more than 350 million people chronically infected and at risk of hepatic decompensation, cirrhosis, and hepatocellular carcinoma. An improved understanding of hepatitis B virology, immunology, and the natural course of chronic infection, has identified hepatitis B virus replication as the key driver of immune-mediated liver injury and disease progression. The approval of potent oral antiviral agents has revolutionised hepatitis B treatment since 1998. Conventional and pegylated interferon alfa and nucleoside and nucleotide analogues are widely authorised treatments, and monotherapy with these drugs greatly suppresses virus replication, reduces hepatitis activity, and halts disease progression. However, hepatitis B virus is rarely eliminated, and drug resistance is a major drawback during long term therapy. The development of new drugs and strategies is needed to improve treatment outcomes. PMID- 19217992 TI - Mechanical supports for acute, severe ankle sprain: a pragmatic, multicentre, randomised controlled trial. AB - BACKGROUND: Severe ankle sprains are a common presentation in emergency departments in the UK. We aimed to assess the effectiveness of three different mechanical supports (Aircast brace, Bledsoe boot, or 10-day below-knee cast) compared with that of a double-layer tubular compression bandage in promoting recovery after severe ankle sprains. METHODS: We did a pragmatic, multicentre randomised trial with blinded assessment of outcome. 584 participants with severe ankle sprain were recruited between April, 2003, and July, 2005, from eight emergency departments across the UK. Participants were provided with a mechanical support within the first 3 days of attendance by a trained health-care professional, and given advice on reducing swelling and pain. Functional outcomes were measured over 9 months. The primary outcome was quality of ankle function at 3 months, measured using the Foot and Ankle Score; analysis was by intention to treat. This study is registered as an International Standard Randomised Controlled Trial, number ISRCTN37807450. RESULTS: Patients who received the below knee cast had a more rapid recovery than those given the tubular compression bandage. We noted clinically important benefits at 3 months in quality of ankle function with the cast compared with tubular compression bandage (mean difference 9%; 95% CI 2.4-15.0), as well as in pain, symptoms, and activity. The mean difference in quality of ankle function between Aircast brace and tubular compression bandage was 8%; 95% CI 1.8-14.2, but there were little differences for pain, symptoms, and activity. Bledsoe boots offered no benefit over tubular compression bandage, which was the least effective treatment throughout the recovery period. There were no significant differences between tubular compression bandage and the other treatments at 9 months. Side-effects were rare with no discernible differences between treatments. Reported events (all treatments combined) were cellulitis (two cases), pulmonary embolus (two cases), and deep-vein thrombosis (three cases). INTERPRETATION: A short period of immobilisation in a below-knee cast or Aircast results in faster recovery than if the patient is only given tubular compression bandage. We recommend below-knee casts because they show the widest range of benefit. FUNDING: National Co ordinating Centre for Health Technology Assessment. PMID- 19217994 TI - Collapse while jogging--and a mimic of milk. PMID- 19217995 TI - [Pneumococcal disease and emergence of serotypes in the vaccine era. Transient trends and serotype replacement?]. PMID- 19217996 TI - [Corynebacterium urealyticum: from the bedside to complete genome sequence]. PMID- 19217997 TI - [Assessment of factors associated with patients' comprehension of treatment at the start of antiretroviral therapy]. AB - OBJECTIVE: The aim of this study was to evaluate factors associated with patients' comprehension of antiretroviral therapy (ART). METHOD: Cross-sectional analysis in which patients at 2 HIV/AIDS public referral centers (Belo Horizonte, Brazil) were interviewed after initiating ART. Information was recorded on variables related to the patient's characteristics, the treatment prescribed, and the healthcare professional involved. A score indicating the patients' level of comprehension regarding the medications prescribed was obtained using a latent trait model estimated by the item response theory. RESULTS: A total of 406 patients were interviewed. Mean (SD) age was 35 (10) years, 227 were men (56%), 302 of Afro-American ethnicity (77%), and 213 had <8 years of education (53%). The regression model determined that 52.25% of the variability of comprehension was explained by the individual's characteristics. Variables associated (P<0.05) with poorest understanding about ART were lower education (<8 years), lack of knowledge about treatment duration and clinical severity, inadequate information provided by physicians, inability to understand pharmaceutical information, daily number of tablets, and the ART regimen prescribed. CONCLUSION: Comprehension of information about the ART regimen prescribed varies considerably between individuals. Nonetheless, several factors were found to be associated with the level of understanding: characteristics of the patient (education, clinical severity), characteristics of treatment (daily number of tablets, ART regimen prescribed), and contribution of healthcare professionals (information from physicians and pharmacists). Strategies to reinforce information about ART should be a priority for patients with a low level of understanding. PMID- 19217999 TI - [Infectious complications in 159 consecutive kidney transplant recipients]. AB - INTRODUCTION: The incidence of infections has decreased in kidney transplant (KT) recipients owing to advances in the surgical techniques and clinical management of this population. Nevertheless, these complications continue to occur and the causes seem to be changing, in part because of the prophylactic strategies used. METHOD: Prospective, observational study investigating infections occurring during the first 2 years post-transplantation in KT recipients who underwent surgery between July 2003 and December 2005 at Hospital Universitario Virgen del Rocio. Univariate and multivariate regression analysis was performed to determine risk factors associated with the development of infection. RESULTS: The incidence of infection was 1.11 episodes per patient over 510+/-234 days. The most common infections were urinary tract infection (UTI) (46.6%), cytomegalovirus (CMV) infection (22.7%), and surgical site infection (8%). The causes were bacterial (50.4%), viral (45.9%), and fungal (3.6%) agents. The most frequent pathogens were CMV (36%), Escherichia coli (28%), extended-spectrum beta-lactamase (ESBL) producers (26%), and coagulase-negative staphylococci (6.3%). Seventy-nine percent of infection episodes occurred in the 4 months following KT. One recipient died 30 days after the infection episode. In the infection group, patient and graft survival at the end of follow-up was 98% and 89%, respectively. CONCLUSIONS: The most frequent syndromes were UTI, CMV infection and surgical site infection. The infections were mainly produced by bacteria, in particular gram-negative rods, and there was a high rate of ESBL E. coli. PMID- 19217998 TI - [Epidemiological features of invasive pneumococcal disease before and after the introduction of pneumococcal conjugate vaccine in Gran Canaria (Canary Islands)]. AB - INTRODUCTION: The introduction of pneumococcal conjugate vaccine (PCV-7) has modified the epidemiology of invasive pneumococcal disease (IPD). Our aim was to investigate the epidemiological features of IPD before and after implementing the use of PCV-7. METHODS: All invasive Streptococcus pneumoniae strains isolated in our hospital from 2000 to 2006 were included. Serotypes were identified and antibiograms were performed in all cases. Data obtained before (2000-2001) and after (2004-2006) authorization of PCV-7 use in Spain were compared. RESULTS: There were 241 cases of IPD. None of the patients with IPD aged 2 years or younger had received PCV-7. PCV-7 coverage in children aged 5 or younger was about 48%. There was a non-significant increase in the incidence of IPD in children (from 53.8 to 57.8 cases/100 000 population), with no change in adults. When IPD incidence was adjusted by the number of blood samples collected, there was a non-significant decrease in both children and adults. Since PCV-7 came on the market, there has been a decrease in vaccine serotypes and an increase in non vaccine serotypes. The emergent serotypes since that time include 3, 6A, 15, and 19A. Penicillin resistance decreased significantly (p<0.001) from the pre-vaccine period (87.3%) to 2003 (13.8%), and later rose from 2003 through 2006 (41.7%). Erythromycin resistance showed no changes during the study. CONCLUSIONS: The incidence of IPD in children aged 2 years and younger in Gran Canaria has not decreased despite the introduction of PCV-7. However, there has been a reduction in the number of cases related to vaccine serotypes and a significant decrease in penicillin resistance. In contrast, non-vaccine serotype IPD cases have increased. Universal vaccination and the use of new polyvalent vaccines may enhance these effects. PMID- 19218001 TI - Community-acquired Capnocytophaga canimorsus meningitis in adults: report of one case with a subacute course and deafness, and literature review. AB - INTRODUCTION: An adult with community-acquired Capnocytophaga canimorsus meningitis presented with a subacute course, deafness, and a predominantly lymphocytic CSF cell count. METHODS: Eighteen reported cases of C. canimorsus meningitis in adults were analyzed. RESULTS: In about half the cases, CSF contained less than 1,000 leukocytes/microL or lymphocyte percentages 30%. These figures differ from those usually seen in meningitis caused by the classic meningeal pathogens. CONCLUSIONS: C. canimorsus should be included among the causes of sporadic lymphocytic meningitis in adults, in particular if the presentation involves deafness. PMID- 19218002 TI - [Evolutionary engineering in Salmonella: emergence of hybrid virulence-resistance plasmids in non-typhoid serotypes]. AB - An example of evolutive engineering in bacterial pathogens is the emergence of hybrid virulence-resistance (VR) plasmids in Salmonella enterica, resulting from an association between antimicrobial resistance determinants and specific virulence plasmids of the S. typhimurium and S. choleraesuis serotypes. VR plasmids all possess the spv (Salmonella plasmid virulence) operon, which is involved in systemic infection; however, they differ in the presence of other virulence determinants and in the resistance gene profile. VR plasmids of S. typhimurium have been found in Europe, and show resistance regions with different levels of complexity that can include class 1 integrons and various transposons. VR plasmids of S. choleraesuis, detected in strains isolated in Taiwan, only confer resistance to ampicillin and sulfonamides. Both serotypes are zoonotic and the presence of hybrid VR plasmids may confer an adaptive advantage under certain conditions, resulting in bacterial strains that are more difficult to treat and have a higher epidemic potential. PMID- 19218003 TI - [Antimicrobial mechanisms of action]. AB - A large number of families and groups of antimicrobial agents are of clinical interest. The mechanisms by which compounds with antibacterial activity inhibit growth or cause bacterial death are varied and depend on the affected targets. The bacterial cell wall-a unique structure in most bacteria that is absent in eukaryotic cells-can be affected in several ways: at different stages of synthesis (fosfomycin, cycloserine) or transport (bacitracin, mureidomycins) of its metabolic precursors, or by a direct action on its structural organization (beta-lactams, glycopeptides). The main drugs affecting the cytoplasmic membrane are polymyxins and daptomycin. Protein synthesis can be blocked by a large variety of compounds that affect any of the phases of this process, including activation (mupirocin), initiation (oxazolidinones, aminoglycosides), binding of the tRNA amino acid complex to ribosomes (tetracyclines, glycylcyclines) and elongation (amphenicols, lincosamides, macrolides, ketolides, streptogramins, fusidic acid). The metabolism of nucleic acids can be altered at the DNA dependent RNA polymerase or in the process of DNA coiling (quinolones); some compounds affect DNA directly (nitroimidazoles, nitrofurans). Trimethoprim and sulfamides (often used in combination) are examples of antimicrobial agents that block bacterial metabolic pathways. Some compounds are unable to inhibit or kill bacteria in themselves, but can block bacterial mechanisms of resistance, enhancing the activity of other antimicrobials administered in combination. Among this group of agents, only certain beta-lactamase inhibitors are currently in clinical use. PMID- 19218005 TI - [Shepherd with fever of 2 weeks of evolution]. PMID- 19218004 TI - [Long evolution of diarrhea with a debatable infectious etiology in a homosexual patient infected with HIV]. PMID- 19218000 TI - Fosamprenavir (GW433908)/ritonavir in HIV-infected patients: efficacy and safety results from the Spanish Expanded Access Program. AB - INTRODUCTION: The use of protease inhibitors (PI) has led to a decrease in HIV-1 related mortality and morbidity. The objective of this study was to collect safety data on treatment with fosamprenavir/ritonavir (FPV/r) 700/100mg BID in HIV-infected patients through an expanded access program. PATIENTS AND METHODS: Prospective, multicenter, noncomparative study in HIV-1 infected adults, for whom a regimen containing FPV/r 700/100mg BID was appropriate. RESULTS: A total of 678 patients were included in the intention-to-treat (ITT) and safety population. The on-treatment (OT) population contained 587 patients: 76% male, 98% Caucasian, and median age 41 years. Median CD4 cell count was 351 cells/microL, HIV-RNA was 3 log copies/mL, and 49% of patients were in CDC class C. After 24 weeks of treatment, serum viral load decreased a median of 1.3 log copies/mL and 73% of patients had <400 copies/mL (P<.0001 vs. baseline); 48-week results were similar. CD4 cell count increased a median of 49 and 62 cells/microL at 24 and 48 weeks, respectively. Adverse events (AEs) associated with the study medication occurred in 21% of patients. CONCLUSIONS: Ritonavir-boosted fosamprenavir as part of antiretroviral therapy is a potent, safe treatment in real-life clinical circumstances. PMID- 19218006 TI - [Mycobacterium neoaurum bacteremia in an immunodepressed patient]. PMID- 19218007 TI - [Activity of mupirocin against methicillin-resistant Staphylococcus aureus isolated in Pontevedra province]. PMID- 19218008 TI - [Chorea during treatment with pegylated interferon and ribavirin in HIV-HCV coinfected patient]. PMID- 19218009 TI - [Serotypes of Streptococcus pneumoniae isolated from children with acute otitis media]. PMID- 19218010 TI - [Pulmonary malacoplakia associated with Rhodococcus equi infection in a patient with AIDS]. PMID- 19218012 TI - Increase of hematopoietic progenitor and suppression of endothelial gene expression by Runx1 expression during in vitro ES differentiation. AB - OBJECTIVE: Runx1 is essential for both the establishment of hematopoiesis during development and maintenance of adult hematopoiesis. To reveal the roles of Runx1, we examined how and when Runx1 functions during development of hematopoiesis, and revealed the genes controlled by Runx1. MATERIALS AND METHODS: A combined in vitro approach involving in vitro hematopoietic differentiation of embryonic stem cells and conditional gene expression of Runx1 was utilized for this study. Then we analyzed the effects of Runx1 on the differentiation and proliferation of hematopoietic cells and carried out DNA microarray analysis. RESULTS: Pulse expression of Runx1 prior to the emergence of hematopoietic cells caused immature hematopoietic cell increase but did not have any effects on the induction of hemogenic cells. During this process, the mRNA level of several endothelial cell specific genes was downregulated. CONCLUSION: Runx1 expression play important roles on the proliferation of emerging immature hematopoietic progenitors or the transition process from endothelial to hematopoietic cells presumably by suppressing the genes related to endothelial phenotype. PMID- 19218011 TI - Nonfucosylated rituximab potentiates human neutrophil phagocytosis through its high binding for FcgammaRIIIb and MHC class II expression on the phagocytotic neutrophils. AB - OBJECTIVE: Antibody-dependent cellular cytotoxicity mediated by natural killer cells via leukocyte receptor IIIa (FcgammaRIIIa) is greatly enhanced by the absence of the core fucose of Fc oligosaccharides, and is closely related to the clinical efficacy of anticancer processes in humans in vivo. Here, we focused on the physiological functions of nonfucosylated anti-CD20 IgG1 rituximab, in particular those functions mediated by human neutrophils, which highly express FcgammaRIIIb, a highly homologous FcgammaR to FcgammaRIIIa. MATERIALS AND METHODS: After treatment with anti-CD20, the response of neutrophils to fluorescently labeled CD20(+) B-cell lymphoma in human whole blood was quantitatively analyzed by measuring their activities of antibody-dependent phagocytosis and major histocompatibility complex (MHC) class II expression on the phagocytotic neutrophils using flow cytometry. RESULTS: In human whole blood, most of the added CD20(+) B-cell lymphoma died shortly, within 4 hours, irrespective of the presence or absence of anti-CD20. Neutrophils were not directly concerned in the death because depletion of neutrophils from human whole blood did not affect the phenomenon. However, neutrophils aggressively phagocytosed newly dead lymphoma cells, and the nonfucosylated anti-CD20 effectively enhanced neutrophil phagocytosis solely by enhancing binding for the phagocytosis coreceptor FcgammaRIIIb. Noteworthy, more increased expression of MHC class II was also observed on the phagocytotic neutrophils than those observed on spontaneous and fucosylated anti-CD20 stimulated phagocytotic neutrophils. CONCLUSIONS: Our data showed that antibody therapy composed of nonfucosylated rituximab can activate human neutrophil functions involving phagocytosis and MHC class II expression, which may favorably potentiate the adaptive immune response in cancer patients. PMID- 19218013 TI - The brain has long been considered the "black box" of the human body. Foreword. PMID- 19218014 TI - Neurological emergencies are a common presentation. Preface. PMID- 19218015 TI - Rapid focused neurological assessment in the emergency department and ICU. AB - In the emergency and critical care setting, a comprehensive and thorough neurologic examination can be impractical. The clinical context should therefore focus the examination on those features relevant to acute diagnosis and management. This article discusses how to direct the history and examination in patients who have focal complaints, possible strokes affecting the anterior or posterior circulations, neck or back pain, neuromuscular complaints, global symptoms, or nonanatomic complaints. PMID- 19218016 TI - Critical care transport of patients who have acute neurological emergencies. AB - This article reviews the special questions and issues in critical care transport related specifically to the care of patients who have neurologic emergencies. It first considers potential indications for transport and reviews attempts to create a hierarchical stroke center system akin to that developed for trauma care. It then discusses therapeutic concerns relating to the transport environment and the use of specific interventions, including the effects of end tidal CO(2) monitoring on intracranial pressure, patient outcomes after traumatic brain injury, and opportunities to initiate therapeutic hypothermia in comatose survivors of cardiac arrest during transport. Finally, the cost of critical care transport of patients who have neurologic emergencies is considered. PMID- 19218017 TI - Clinical nihilism in neuroemergencies. AB - Mortality and morbidity remain high from neurologic emergencies, such as acute stroke, traumatic brain injury, and hypoxic-ischemic encephalopathy after cardiac arrest. Decisions regarding initial aggressiveness of care must be made at the time of presentation, and perceived prognosis is often used as part of this decision-making process. These decisions are predicated on the accuracy of early outcome prediction, however. Decisions to limit treatment early after neuroemergencies must be balanced with avoidance of self-fulfilling prophecies of poor outcome attributable to clinical nihilism. This article examines the role of prognostication early after neuroemergencies, the potential impact of early treatment limitations, and how these may relate to communication with patients and surrogate decision makers in the context of these acute neurologic events. PMID- 19218018 TI - Vertigo and dizziness in the emergency department. AB - Understanding three peripheral vestibular disorders--vestibular neuritis, benign paroxysmal positional vertigo, and Meniere's disease--is the key to the evaluation and management of vertigo and dizziness presentations in the emergency department. Each of these benign disorders is a common cause of a broad category of dizziness presentation. In addition, each of these disorders has characteristic features that allow for a bedside diagnosis. An effective strategy for "ruling-out" a serious disorder, such as stroke, is "ruling-in" a peripheral vestibular disorder. In this article a focus is on the key features of these disorders. PMID- 19218019 TI - Management of transient ischemia attacks in the twenty-first century. AB - Patients who have a transient ischemic attack (TIA) represent a group that may appear well but are at high risk for stroke within 90 days. Management of patients who have a TIA requires an understanding of the short-term risk for stroke to guide acute management and the long-term benefits to medical and surgical therapies. The initial emergency department (ED) evaluation may be supplemented with simple TIA stroke risk scores to estimate short-term stroke risk. The addition of MRI provides yet more information regarding stroke risks, while identifying some specific causes of TIA. Extended testing may not be feasible in the ED, and to address this limitation, new outpatient strategies for patients who have a TIA have been developed, such as the use of an ED observation unit or an outpatient TIA clinic. Although controversy remains in some areas of acute TIA management because of the lack of evidence from controlled trials, evidence from large randomized trials have led to a better understanding of effective measures for the long-term prevention of stroke. PMID- 19218021 TI - Central nervous system infections. AB - Central nervous system infections have long been recognized as among the most devastating of diseases. This article describes the changing pattern and epidemiology of a variety of common central nervous system infections, including meningitis, encephalitis, and brain abscesses, and reviews pathophysiology and the most current approach to clinical diagnosis, treatment, and disposition from the emergency physician perspective. PMID- 19218020 TI - Diagnosis and management of the primary headache disorders in the emergency department setting. AB - Headache continues to be a frequent cause of emergency department (ED) use, accounting for 2% of all visits. Most of these headaches prove to be benign but painful exacerbations of chronic headache disorders, such as migraine, tension type, and cluster. The goal of ED management is to provide rapid and quick relief of benign headache, without causing undue side effects, and to recognize headaches with malignant course. Although these headaches have distinct epidemiologies and clinical phenotypes, there is overlapping response to therapy; nonsteroidals, triptans, dihydroergotamine, and the antiemetic dopamine antagonists may play a therapeutic role for each of these acute headaches. This article reviews the diagnostic criteria and management strategies for the primary headache disorders. PMID- 19218022 TI - Emergency treatment of status epilepticus: current thinking. AB - Current thinking about the acute treatment of status epilepticus (SE) emphasizes a more aggressive clinical approach to this common life-threatening neurologic emergency. In this review, the authors consider four concepts that can accelerate effective treatment of SE. These include (1) updating the definition of SE to make it more clinically relevant, (2) consideration of faster ways to initiate first-line benzodiazepine therapy in the prehospital environment, (3) moving to second-line agents more quickly in refractory status in the emergency department, and (4) increasing detection and treatment of unrecognized nonconvulsive SE in comatose neurologic emergency patients. PMID- 19218024 TI - Therapeutic hypothermia for neuroprotection. AB - This review briefly discusses induced therapeutic hypothermia (TH), which represents the intentional induction of a lowered core body temperature of 35 degrees C or less. The focus is on resuscitative or postarrest hypothermia, the data that support it, and the practical issues pertaining to TH implementation. PMID- 19218025 TI - Glycemic control and the injured brain. AB - The interaction between glycemic control and critical neurologic illness and injury is complex. Hyperglycemia can be either the cause or the result of severe brain injury. Hyperglycemia in acute neurologic injury is associated with worse neurologic outcomes. Demographic patterns, including an aging population and shifts in racial and ethnic representation, contribute to the increasing prevalence of hyperglycemia and diabetes among victims of the most common neurologic emergencies. This article reviews the epidemiology of the problem, relevant pathophysiology, the use of tight glycemic control therapy in other populations, and the potential for tight glycemic control as a way to improve outcomes after acute neurologic illness and injury. PMID- 19218026 TI - Controversies and advances in neonatal neurology: overview. Introduction. PMID- 19218023 TI - Enhancing community delivery of tissue plasminogen activator in stroke through community-academic collaborative clinical knowledge translation. AB - Improving the clinical outcomes of stroke patients depends on the adoption of proven new therapies throughout the broader medical community. Approximately 1% of stroke patients in community settings are receiving tissue plasminogen activator (tPA) therapy 12 years after US Food and Drug Administration approval. Knowledge translation, the process by which the results of clinical investigations are adopted by clinicians and incorporated into routine practice, is important but often overlooked. This article reviews the history of tPA use in stroke as a case study of a breakdown of knowledge translation in emergency medicine. It reviews knowledge translation concepts and theory and explores practical community-academic collaborative methods based on these tenets to enhance acute stroke care delivery in the community setting. PMID- 19218027 TI - Ethics in neonatal neurology: when is enough, enough? AB - Decision-making for the severely neurologically compromised newborn is fraught with considerable medical and ethical difficulties. Multiple intrinsic and extrinsic factors challenge our prognostic certainty, which then challenges our ability both to communicate and to make decisions that are rooted in basic principles of fairness and moral integrity. Clinical vignettes illustrate the difficulties and highlight the present status regarding prognostication and consideration of general ethical principles that would permit the consensual withdrawal of care (i.e., end-of-life decisions). Futility in the neonatal setting is examined, as well the mechanisms of the decision-making process and existing relevant professional guidelines. PMID- 19218028 TI - Animal models of perinatal hypoxic-ischemic brain damage. AB - Animal models are often presumably the first step in determining mechanisms underlying disease, and the approach and effectiveness of therapeutic interventions. Perinatal brain damage, however, evolves over months of gestation, during the rapid maturation of the fetal and newborn brain. Despite marked advances in our understanding of these processes and technologic advances providing an improved window on the timing and duration of injury, neonatal brain injury remains a "moving target" regarding our ability to "mimic" its processes in an animal model. Moreover, interfering with normal processes of development as part of a therapeutic intervention may do "more harm than good." Hence, controversy continues over which animal model can reflect human disease states. Numerous models have provided information regarding the pathophysiology of brain damage in term and preterm infants. Our challenges consist of identifying infants at greatest risk for permanent injury, identifying the timing of injury, and adapting therapies that provide more benefit than harm. A combination of appropriately suitable animal models to conduct these studies will bring us closer to understanding human perinatal damage and the means to treat it. PMID- 19218029 TI - Role of perinatal inflammation in cerebral palsy. AB - Inflammatory molecules are promptly upregulated in the fetal environment and postnatally in brain-damaged subjects. Intrauterine infections and inflammation are often associated with asphyxia. This double-hit effect by combined infection or inflammation and hypoxia is therefore a frequent concomitant in neonatal brain damage. Animal models combining hypoxia and infection were recently designed to explore the mechanisms underlying brain damage in such circumstances and to look for possible neuroprotective strategies. Proinflammatory cytokines are thought to be major mediators in brain injury in neonates with perinatal asphyxia, bacterial infection, or both. Cytokines, however, could also have neuroprotective properties. The critical point in the balance between neurodamaging and neuroprotective effects of cytokines has yet to be unraveled. This understanding might help to develop new therapeutic approaches to counteract the inflammatory disequilibrium observed in the pathophysiologic mechanisms associated with brain injury. PMID- 19218030 TI - Neonatal seizures: do they damage the brain? AB - Seizures are an early sign of brain injury in newborns. These seizures are in most cases repetitive or associated with asymptomatic electrographic seizures. Despite the relative resistance of the immature brain to seizure-induced brain damage, there is more and more evidence that neonatal seizures impair normal brain development. This review addresses the changes associated with neonatal seizures and discusses current and future potential neuroprotective strategies. PMID- 19218031 TI - Advanced neuroimaging techniques for the term newborn with encephalopathy. AB - Neonatal encephalopathy is associated with a high risk of morbidity and mortality in the neonatal period and of long-term neurodevelopmental disability in survivors. Advanced magnetic resonance techniques now play a major role in the clinical care of newborns with encephalopathy and in research addressing this important condition. From conventional magnetic resonance imaging, typical patterns of injury have been defined in neonatal encephalopathy. When applied in contemporary cohorts of newborns with encephalopathy, the patterns of brain injury on magnetic resonance imaging distinguish risk factors, clinical presentation, and risk of abnormal outcome. Advanced magnetic resonance techniques such as magnetic resonance spectroscopy, diffusion-weighted imaging, and diffusion tensor imaging provide novel perspectives on neonatal brain metabolism, microstructure, and connectivity. With the application of these imaging tools, it is increasingly apparent that brain injury commonly occurs at or near the time of birth and evolves over the first weeks of life. These observations have complemented findings from trials of emerging strategies of brain protection, such as hypothermia. Application of these advanced magnetic resonance techniques may enable the earliest possible identification of newborns at risk of neurodevelopmental impairment, thereby ensuring appropriate follow-up with rehabilitation and psychoeducational resources. PMID- 19218032 TI - Outcomes for the extremely premature infant: what is new? And where are we going? AB - Two approaches were taken to determine what is new and where we are going in terms of outcomes for the extremely premature infant: publications from 2004 to 2007 were reviewed, and the 30-year outcome at the authors' institutions was assessed. Recent literature documents improving early childhood outcomes in the face of improved survival. Childhood cerebral palsy prevalence rates have been reported to be as low as 19 per 1000 live births for infants born at 20-27 weeks gestation. Vision and hearing loss have been reported in fewer than 1% of survivors. The rate of overall intellectual impairment has not improved, although impairment was reduced in a recent trial of caffeine therapy for apnea of prematurity, and this remains an important area for study. In sum, recent findings herald a more positive perspective on the outcome for extremely premature survivors. It can thus be expected that new intensive-care trials will attempt to reduce the proportion of survivors with adverse outcomes. Childhood assessments will have a greater focus on function and participation. Information on improved outcomes for preterm infants will inform guidelines of decision making used to help parents to determine what is best for their child. The audit component of follow-up studies will expand and more cohort and trial studies will become multicenter, national, and international. PMID- 19218033 TI - A new look at outcomes of infants with congenital heart disease. AB - This article provides an overview of a longitudinal study on a cohort of 131 newborns and young infants with congenital heart defects who required open heart surgery. The rationale for the study design is provided as well as a summary of the procedures used to evaluate these children prior to surgery, at discharge after surgery, 12-18 months later, and at 5 years of age. Results demonstrate that a substantial proportion of these infants had neurologic and developmental abnormalities prior to surgical repair. Developmental delays were common in children with both cyanotic and acyanotic heart defects, and these deficits persisted to school entry. A number of medical, surgical, demographic and environmental factors were significantly associated with developmental outcomes suggesting a multifactorial etiology to brain injury. Limited educational and rehabilitation resources for this cohort at early school age suggests that service needs may need to be more carefully planned for this high-risk population. Lessons learnt from this prospective study are highlighted as well as future directions for research and clinical practice. PMID- 19218034 TI - Advances in perinatal ischemic stroke. AB - Increasingly distinct patterns of focal ischemic injury in the fetal and perinatal brain are recognized. Improved classification has afforded advances in risk factor identification, pathophysiology hypotheses, outcome prediction, and potential avenues for intervention. Cerebrovascular occlusion leading to perinatal stroke may be arterial or venous, symptomatic or subclinical, and it can occur across multiple time frames. Distinguishing causative factors from mere associations represents a major challenge with important implications for studies of pathogenesis. The adverse outcomes suffered by most children highlight the need for further research. Reviewed here are the current understandings, recent advancements, and future directions for research in perinatal ischemic stroke. PMID- 19218035 TI - Systematic review of biomarkers of brain injury in term neonatal encephalopathy. AB - Although neonatal hypoxic-ischemic encephalopathy is a common cause of childhood developmental disability, its timing, duration, and outcomes are poorly defined. Biomarkers serve as surrogates for disease injury, evolution, and outcome, but no tissue biomarker in routine clinical use can help predict outcomes in term newborn encephalopathy. We reviewed biomarkers in human term neonatal encephalopathy, to determine if current biomarkers are strong enough for clinical use as predictors of outcomes. A comprehensive search of databases identified 110 publications that met our inclusion criteria, i.e., (1) newborns at >36 weeks; (2) neonatal encephalopathy as defined by the American College of Obstetrics and Gynecology; (3) the use of a serum, urine, or cerebrospinal fluid biomarker; and (4) reported outcomes beyond age 12 months. Of those 110 publications, 22 reported outcomes beyond age 12 months. In single reports, urine lactate (P < 0.001), first urine S100 (P < 0.0001), cord-blood interleukin-6 (P = 0.02), serum nonprotein-bound iron (P < 0.001), serum CD14 cell NFkappaB activation (P = 0.014), serum interleukin-8 (P = 0.03), and serum ionized calcium (P = 0.001) were potential predictors of death or abnormal outcomes. A meta-analysis identified serum interleukin-1b (P = 0.04, n = 3), serum interleukin-6 (P = 0.04, n = 2), cerebrospinal fluid neuron-specific enolase (P = 0.03, n = 3), and cerebrospinal fluid interleukin-1b (P = 0.003, n = 2) as putative predictors of abnormal outcomes in survivors, when measured before age 96 hours. Several serum, urine, and cerebrospinal fluid biomarkers of term neonatal encephalopathy may provide important information regarding long-term outcomes. None, however, were studied extensively enough to warrant routine clinical use. Validation of these markers, either alone or in combination, is required in the development of viable therapeutic interventions. PMID- 19218036 TI - Neuroimaging as a basis for rational stem cell therapy. AB - Neonatal global or focal hypoxic-ischemic brain injury remains a frequent and devastating condition, with serious long-term sequelae. An important issue in any neonatal clinical trial of neuroprotective agents relates to developing accurate measures of injury severity and also suitable measures of the response to treatment. Advanced magnetic resonance imaging techniques can acquire serial and noninvasive data about brain structure, metabolic activity, and the response to injury or treatment. These imaging methods need validation in appropriate animal models for translational research studies in human newborns. This review describes several approaches that use imaging as well as proton magnetic resonance spectroscopy to assess the severity of ischemic injury (e.g., for possible candidate selection) and for monitoring the progression and evolution of injury over time and as an indicator of recovery or response to treatment. Preliminary data are presented on how imaging can be used after neural stem cell implantation to characterize the migration rate, the magnitude of stem cell proliferation, and their final location. Imaging has the potential to allow monitoring of many dimensions of neuroprotective treatments and can be expected to contribute to efficacy and safety when clinical trials using neural stem cells or other neuroprotective agents become available. PMID- 19218037 TI - Treatment of the term newborn with brain injury: simplicity as the mother of invention. AB - Neonatal brain injury remains a common cause of developmental disability, despite tremendously enhanced obstetrical and neonatal care. The timing of brain injury occurs throughout gestation, labor, and delivery, providing an evolving form of brain injury and a moving target for therapeutic intervention. Nonetheless, markedly improved methods are available to identify those infants injured at birth, via clinical presentation with neonatal encephalopathy and neuroimaging techniques. Postischemic hypothermia has been shown to be of tremendous clinical promise in several completed and ongoing trials. As part of this approach to the treatment of the newborn, other parameters of physiologic homeostasis can and should be attended to, with strong animal and clinical evidence that their correction will have dramatic influence on the outcome of the newborn infant. This review addresses aspects of newborn care to which we can direct our attention currently, and which should result in a safe and efficacious improvement in the prognosis of the newborn with neonatal encephalopathy. PMID- 19218038 TI - The Neuroscience Peer Review Consortium. PMID- 19218039 TI - Association between serum albumin and mortality in dialysis patients is partly explained by inflammation, and not by malnutrition. AB - OBJECTIVE: We investigated the effects of inflammatory and nutritional status on the association between serum albumin and mortality in hemodialysis (HD) and peritoneal dialysis (PD) patients. DESIGN AND PATIENTS: This was a prospective cohort study of incident dialysis patients starting HD or PD. Inflammation (C reactive protein >or=5 or >or=10 mg/L), malnutrition (1 to 5 on the 7-point subjective global assessment [SGA]), and low protein intake (normalized protein equivalent of nitrogen appearance [nPNA] <0.99 g/kg/day) were measured at 3 months after the start of dialysis. SETTING: The study involved 38 dialysis centers in The Netherlands. MAIN OUTCOME MEASURE: We ascertained all-cause mortality during the first 2 years after the start of dialysis. RESULTS: In total, 700 patients were included (mean SD age, 59 [+/-15] years; serum albumin, 3.3 (0.7) g/dL; 60% men; 454 starting HD, and 246 starting PD). The 2-year mortality was 21%. In HD patients, the mortality (hazard ratio [HR], with 95% confidence interval [95% CI]) per unit decrease in serum albumin (g/dL) was 1.47 (95% CI, 1.07 to 2.00). Adjustment for SGA did not decrease this risk, whereas adjustment for nPNA decreased the HR to 1.45 (95% CI, 1.06 to 1.97). The mortality risk decreased to 1.30 (95% CI, 0.95 to 1.78) after adjustment for inflammation, and did not further decrease after additional adjustment for SGA and nPNA. Additional adjustments for age, sex, and comorbidity decreased the HR to 1.09 (95% CI, 0.79 to 1.51). In PD patients, the effects of adjustments on the mortality risk of serum albumin (1.38; 95% CI, 0.87 to 2.20) were similar. CONCLUSION: In dialysis patients, a 1-g/dL decrease in serum albumin was associated with an increased mortality risk of 47% in HD patients and 38% in PD patients. These mortality risks were in part explained by the inflammatory pathway. The mortality risks associated with serum albumin were not a consequence of malnutrition, as measured with SGA and nPNA. These findings imply that nutritional status cannot be assessed with precision by the measurement of serum albumin in dialysis patients. PMID- 19218040 TI - Implementation of standardized nutrition guidelines by renal dietitians is associated with improved nutrition status. AB - OBJECTIVE: Standardized nutrition guidelines that focus on a nutrition care process have been used by dietitians treating renal patients in Australia for over 3 years. We show the impact of this implementation on the nutritional status of a cohort of hemodialysis patients. DESIGN: We conducted a retrospective observational study, investigating a cohort of maintenance hemodialysis patients after the implementation of a systematic approach to the patient's nutritional care. SETTING: This study took place in public and private in-center hemodialysis units. PATIENTS: Patients included a cohort of 65 maintenance hemodialysis patients (mean age +/- SD, 64 +/- 15 years; 58% male; dialysis vintage median [interquartile range], 22 [10 to 46] months). INTERVENTIONS: All participants were provided with a dietary interview at least every 6 months, with intensive follow-up where required, and were monitored monthly regarding weight and biochemistry. Outcomes were assessed annually between May 2004 and December 2006, after the implementation of this model of care. MAIN OUTCOME MEASURE: Energy and protein intake according to dietary interview, nutritional status according to subjective global assessment, and data regarding dry weight and biochemistry (including albumin, potassium, and phosphate) were collected by the dietitian at each facility. Change in each outcome measure over time was assessed using repeated-measures analysis. RESULTS: The proportion of patients with malnutrition (subjective global assessment B or C) decreased from 14% at baseline to 3% after 2 years. Serum albumin, potassium, and dry weight remained stable throughout the study period, and there was a significant decrease in serum phosphate over time (mean +/- SD,1.8 +/- 0.5 to 1.5 +/- 0.5 mmol/L, P = .004). Dietary energy and protein intake changed significantly over the study period (P = .001 and P = .022, respectively), with the highest mean intake recorded during the final follow-up assessment. CONCLUSIONS: The implementation of a systematic approach to patient care, in line with nutrition management guideline recommendations, was associated with an improvement in nutritional status and dietary intake in this cohort of maintenance hemodialysis patients, without the need for increased resources or dietitian time. PMID- 19218041 TI - Effects of peridialytic oral supplements on nutritional status and quality of life in chronic hemodialysis patients. AB - OBJECTIVE: Our objective was to determine the effects of peridialytic oral supplements on nutritional markers and quality of life (QOL) in patients receiving maintenance hemodialysis. DESIGN: This trial was open, prospective, nonrandomized, and comparative. SETTING: This study was performed at an outpatient hemodialysis unit in a teaching hospital. PATIENTS: This study included 88 adults with chronic kidney disease at stage 5. INTERVENTION: This study involved directly observed nutrition therapy with >or=1 can of enteral nutrition (Nepro) with each hemodialysis session thrice weekly for 3 months, or standard care. MAIN OUTCOME MEASURE: Changes in biochemical markers of nutritional status and QOL, as measured by the Kidney Disease Quality of Life Short Form, were determined. RESULTS: Peridialytic oral nutrition resulted in a significant difference between the nutrition and comparison groups in serum albumin change over time (P = .03; repeated-measures analysis of variance with covariates). Mean (+/-SD) serum albumin concentration did not differ between baseline and month 3 in the nutrition group (3.68 +/- 0.33 g/dL vs. 3.75 +/- 0.40 g/dL; P = .12), but in the comparison group, serum albumin levels declined significantly (3.93 +/- 0.34 g/dL at baseline versus 3.81 +/- 0.37 g/dL at month 3; P = .04). The "role-physical" domain score of the Kidney Disease Quality of Life-Short Form significantly changed over time in the nutrition group versus the comparison group (P = .02; repeated-measures analysis of variance with covariates). Nepro was well-tolerated, and greater than 80% of the prescribed therapy was consumed. CONCLUSION: Oral nutrition, as part of structured, directly observed peridialytic therapy in chronic hemodialysis patients, was well accepted, and resulted in the maintenance of serum albumin levels and QOL with respect to impact of physical health on daily activities. These findings need to be confirmed in a randomized, controlled trial. PMID- 19218042 TI - Behavioral modeling to guide adherence to fluid control in hemodialysis patients. AB - OBJECTIVE: This study used the health belief model (HBM) and the transtheoretical model (TTM) to assess hemodialysis (HD) patients' readiness to comply with recommended fluid intake as it relates to their perceptions of therapy and disease. DESIGN: This was a multicenter, cross-sectional study. SETTING: The setting involved 3 hospital-based HD centers in Lebanon. PATIENTS: Patients included 172 consenting HD patients who had been on HD for at least 3 months and were mentally stable. INTERVENTION: Placement of patients into TTM stages was based on their readiness to comply with recommended fluid intake, as confirmed by 4-week interdialytic weight-gain measures, and taking into account their perceptions toward therapy (using HBM constructs). MAIN OUTCOME MEASURES: Mean scores of HBM constructs for each patient were placed in one of the TTM stages of readiness to change. RESULTS: This study included 54.1% males, the average age was 57.8 +/- 14.0 (SD) years, and average number of years on dialysis was 4.6 +/- 4.9. Regarding stages, 18.5% of participants were placed in precontemplation, 40.0% in contemplation, 38.1% in preparation, and 3.4% were in the action/maintenance stages. The HBM constructs were significantly associated with TTM (P < .01). Perceptions of benefits (P = .04) and self-efficacy (P < .01) were significantly associated with TTM. CONCLUSION: Compliance is a multifaceted topic, essential for the survival and quality of life of HD patients. A better understanding of patients' beliefs and level of self-efficacy is a fundamental key to enhancing patients' compliance to therapy and achieving the required behavioral modifications. PMID- 19218043 TI - The protein equivalent of nitrogen appearance in critically ill acute renal failure patients undergoing continuous renal replacement therapy. AB - OBJECTIVE: To assess the nutritional status of critically ill patients with acute renal failure on continuous renal replacement therapy (CRRT) and their protein needs by estimating the protein equivalent of nitrogen appearance (PNA). DESIGN: Prospective, observational study. SETTING: A 74-bed intensive care unit in a single tertiary care hospital. PATIENTS: Twenty-five consecutive critically ill patients with acute renal failure on CRRT. METHODS: The patients were studied over a period of 24 hours, at initiation on CRRT. The nutritional status was assessed by anthropometry and bioimpedance analysis. The PNA was estimated using the Bergstrom equation and PNA was normalized to body weight. RESULTS: The mean age was 58.2 +/- 17 years and 20 (80%) were male. The mean weight was 67 +/- 12 kg, body mass index was 25 +/- 3.5 kg/m(2), and triceps and subscapular skin fold thickness were 13 +/- 4.6 mm and 15 +/- 2.5 mm, respectively. Bioimpedance studies showed that the total body water was increased at 61.7 +/- 5.5% and body fat was 31.8 +/- 5.4%. The PNA was 103 +/- 35 g/day and normalized PNA was 1.57 +/- 0.4 g/kg/day. The mean protein intake was 0.56 +/- 0.38 g/kg/day, resulting in mean net negative protein balance of 1.0 +/- 0.6 g/kg/day. CONCLUSIONS: Malnutrition was uncommon in patients with acute renal failure at the time of initiation on CRRT, but their total body water was increased. They exhibited hypercatabolism and the mean normalized PNA was 1.57 g/kg/day. A large negative nitrogen balance was observed in them, since their protein intake was suboptimal. PMID- 19218044 TI - Effect of megestrol acetate on serum albumin level in malnourished dialysis patients. AB - OBJECTIVE: Malnutrition is a common problem in dialysis patients, and may affect up to one third of patients. It is associated with high mortality and morbidity. Although a number of studies were performed to determine effective treatment, there is no proven medication for this condition. This study aimed to evaluate the effect of megestrol acetate (MA) on serum albumin levels in malnourished dialysis patients. DESIGN: This was a randomized, controlled clinical trial. SETTING: The setting, a dialysis center at Razi Hospital, Rasht, Iran, provides services to dialysis patients. PATIENTS: Twenty-two malnourished dialysis patients with persistent hypoalbuminemia (albumin, <3.5 g/dL for 2 months) participated, and were randomly assigned to either an experimental group or a control group. INTERVENTION: The experimental group was treated with MA, 40 mg twice daily, over 2 months, and was compared with the control group. The collected data were analyzed using SPSS (version 10; Chicago, IL). RESULTS: After 2 months, the mean (+/-SD) serum albumin level in the experimental group rose from 3.31 +/- 0.31 g/dL to 4.41 +/- 0.31 g/dL, but in the control group, it declined, from 3.35 +/- 0.21 to 3.02 +/- 0.48 g/dL. The difference between the two groups was significant (P = .002). CONCLUSION: At a dose of 40 mg twice a day, MA safely increased serum albumin in malnourished dialysis patients. PMID- 19218045 TI - Correction of metabolic acidosis on serum albumin and protein catabolism in hemodialysis patients. AB - BACKGROUND: The effect of the correction of metabolic acidosis (MA) on serum albumin concentrations (sAlbs) in hemodialysis (HD) patients is controversial. This study evaluated the role of the correction of MA on sAlb concentrations, normalized protein catabolic rate (nPCR), and the effect of the concomitant inflammatory status, in a group of acidotic HD patients. METHODS: The correction of MA by oral supplementation with sodium bicarbonate, and the evaluation of its effect on sAlb, nPCR, and high-sensitivity C-reactive protein (hsCRP), were performed in 29 patients on bicarbonate dialysis for a median of 30 months. Other variables included pre-HD arterial pH, serum bicarbonate, serum creatinine, serum Na, body weight, interdialytic weight gain, pre-HD systolic and diastolic blood pressure, and Kt/V. RESULTS: Serum bicarbonate and pH increased significantly (P < .0001), from 19.1 +/- 0.7 mmol/L to 24.6 +/- 1.1 mmol/L and from 7.33 +/- 0.03 to 7.39 +/- 0.02, respectively (all values with +/- are SD). The nPCR decreased from 1.13 +/- 0.14 g/kg/day to 1.05 +/- 0.14 g/kg/day (P < .0001). The other variables did not change significantly. In 17 patients with high-sensitivity C reactive protein <10 mg/L, sAlb increased from 3.7 +/- 0.3 g/dL to 4.0 +/- 0.3 g/dL (P < .01), whereas in 12 with high-sensitivity C-reactive protein >or=10 mg/L, sAlb did not change (3.5 +/- 0.17 g/dL vs. 3.4 +/- 0.13 g/dL; P = NS). CONCLUSIONS: Oral sodium bicarbonate supplementation is effective in correcting MA in HD patients and does not affect interdialytic weight gain, plasma Na, and blood pressure. The correction of MA is effective in reducing protein catabolism (nPCR) in both inflamed and less inflamed HD patients, but increases sAlb only in patients without inflammation. In inflamed patients, the correction of MA is not sufficient per se to improve sAlb concentrations. PMID- 19218047 TI - Do hemodialysis patients prefer renal-specific or standard oral nutritional supplements? AB - OBJECTIVE: We investigated whether hemodialysis (HD) patients prefer standard or renal-specific oral nutritional supplements (ONS). DESIGN: Standard ONS Fortisip (Nutricia Clinical Care, Wiltshire, Trowbridge, UK) and renal ONS Renilon (Nutricia Clinical Care) and Nepro (Abbott Laboratories, Ltd., Maidenhead, Berkshire, UK) were compared using single-blind taste tests and face-to-face, interviewer-administered questionnaires. SETTING: This study took place in our HD unit in September 2007. PATIENTS: There were 40 patients, including 24 males, 14 smokers, and 26 Caucasians, aged <30 years (n = 6), 31 to 50 years (n = 13), 51 to 70 years (n = 12), and >70 years (n = 9). INTERVENTION: Patients ranked ONS taste on a Likert scale (1 to 5), and compared flavor options, phosphate-binder requirements, and fluid contribution. MAIN OUTCOME MEASURE: Which factors influenced a patient's choice of ONS? RESULTS: Gender, smoking status, ethnicity, and age influenced patients' choices. The taste of Fortisip and Nepro was liked by 58% (n = 23), versus 28% liking Renilon (n = 11). Renilon was disliked by 35% (n = 14), Nepro was disliked by 30% (n = 12), and Fortisip was disliked by 25% (n = 10). The favorite taste was Fortisip, in 52% (n = 21). However, 21% (n = 4) who preferred the taste of renal ONS would not choose them long-term because of their limited flavor ranges. The lack of phosphate binders with Renilon was a deciding factor in 27% (n = 19/33). The low fluid contribution of renal ONS influenced the choice of 43% (n = 12/28). All factors considered, standard ONS remained most popular for patients aged >70 years. However, in all other subgroups, and particularly males and non-Caucasians, renal ONS became more popular. Many patients (23%; n = 9) would sacrifice taste for the benefits of renal ONS. CONCLUSIONS: Renal ONS are more popular in HD patients because of their low fluid contribution and phosphate-binder requirements, which can influence preference over taste. Patients need information to make informed choices. PMID- 19218046 TI - Influence of renal function and diet on acid-base status in chronic kidney disease patients. AB - OBJECTIVE: We investigated the influence of potential renal acid load (PRAL) and renal function on the degree of metabolic acidosis in patients with chronic kidney disease (CKD). DESIGN: This was a cross-sectional study. SETTING: This study was conducted at the Nephrology Outpatient Division of the Hospital Universitario Clementino Fraga Filho (Rio de Janeiro, Brazil). PATIENTS: Thirty CKD patients undergoing conservative treatment were divided according to plasma HCO(3)(-) values into acidotic (HCO(3)(-) 22 mM, n = 15). MAIN OUTCOME MEASURE: Biochemical, nutritional, and anthropometric parameters and PRAL were measured. RESULTS: The mean of plasma HCO(3)(-) values was 17.7 +/- 2.8 mM in the acidotic group, and 25.1 +/- 2.2 mM in the nonacidotic group. There was no significant difference in mean PRAL values between the acidotic (9.8 +/- 6.4 mEq/day) and nonacidotic (12.7 +/- 10.0 mEq/day) groups, but there was a significant correlation between plasma HCO(3)(-) and creatinine clearance (r = 0.78, P < .0001). Based on the receiver operating characteristic curve, the level of creatinine clearance to begin detection of acidosis was 31.8 mL/min, with a sensitivity and specificity of 86.7%. CONCLUSION: The acid-base status of this group of CKD patients undergoing conservative treatment was mainly determined by degree of renal insufficiency rather than diet. PMID- 19218051 TI - Complex varus elbow instability: a terrible triad model. AB - HYPOTHESIS: The terrible triad is a debilitating injury that involves elbow dislocation with injury to the lateral collateral ligament (LCL) complex, the radial head, and the coronoid process. This study investigated the role of these components in varus stability of the elbow. MATERIALS AND METHODS: We investigated the role of these components in varus stability of the elbow using 10 cadaveric fresh frozen upper extremities. The testing order allowed each of four states to be tested (intact, LCL complex deficient or repaired, radial head resected or replaced) at two flexion angles and multiple coronoid resection levels. Values for restraining load (newtons [N]) at 1.5 cm of varus displacement were obtained on a materials testing machine. RESULTS: Beyond a 50% loss of the coronoid process, neither repair of the LCL nor replacing the radial head alone resulted in a statistically significant increase in varus stability. For a loss of the coronoid process between 67-75%, repair of the LCL and replacement of the radial head showed improved stability over repair alone, or radial head replacement alone. For loss of the coronoid beyond 75%, even repair of the LCL and replacement of the radial head did not improve varus stability of the elbow. CONCLUSION: These findings provide a biomechanical basis for aggressive treatment of coronoid fractures as a component of the terrible triad injury. LEVELS OF EVIDENCE: Basic science study. PMID- 19218052 TI - Arthroscopic treatment of posterior impingement of the elbow in athletes: a medium-term follow-up in sixteen cases. AB - HYPOTHESIS: Evaluate the effectiveness of arthroscopic treatment of posterior impingement in the athletes elbow. MATERIALS AND METHODS: 16 elbows were included. An arthroscopic debridement of the posterior fossa of the elbow was performed. All Patients were evaluated preoperatively and after an average of 38 (30-53) months using range of motion, the Modified Andrews Elbow Scoring System (MAESS), VAS in rest and after provocation. RESULTS: The average flexion increased from 138 degrees to 140 degrees . The extension deficit of 8 degrees preoperatively increased to a deficit of average 2 degrees (P < 0.05). The MAESS increased from average preoperative to excellent postoperative (P < 0.05). The average VAS in rest decreased from 3 to 0 and during sporting activities the VAS decreased from 7 to 2 (P < 0.05). DISCUSSION AND CONCLUSION: In this series, arthroscopic debridement of the posterior fossa in athletes with posterior impingement is a procedure that showed excellent mid term results and can therefore be recommended. LEVEL OF EVIDENCE: Level 4; Retrospective case series, no control group. PMID- 19218053 TI - Alterations in function after rotator cuff tears in an animal model. AB - HYPOTHESIS: This study examined the effect of multiple rotator cuff tendon tears on shoulder function in an animal model. MATERIALS AND METHODS: Forty-eight Sprague-Dawley rats were divided into uninjured control, supraspinatus tendon detachment, supraspinatus+infraspinatus tendon detachment, or supraspinatus+subscapularis tendon detachment groups. Functional assessment was determined through ambulatory parameters (paw and stride measures) and range of motion prior to tendon detachment and at various time points after tendon detachment. RESULTS: Ambulatory parameters and total range of motion, representing measures of shoulder function, were significantly altered with rotator cuff tears. The addition of a second torn rotator cuff tendon (infraspinatus or subscapularis)had further detrimental effects on animal shoulder function compared to uninjured control. DISCUSSION: This study demonstrated functional changes in a rat rotator cuff model. Many of the permanent functional changes were likely present because the required motion used for those actions can no longer be performed. For parameters that were transient, compensation with another limb or subsidence of pain may have occurred. CONCLUSION: The findings in this study are consistent with the alterations in shoulder function observed with rotator cuff and other shoulder injuries in the human. Future studies using this model can begin to examine the root of the functional differences, whether it is pain, mechanical deficiency, or a combination of both, which cannot be fully studied clinically [corrected]. PMID- 19218056 TI - Foot & Ankle Surgery enters the world stage of education and research in disorders of the foot and ankle. PMID- 19218054 TI - Treatment of osseous defects associated with anterior shoulder instability. AB - Bone loss of the glenoid and/or humerus is a common consequence of traumatic anterior shoulder instability and can be a cause of recurrent instability after a Bankart repair. Accurate characterization of the size and location of osseous defects associated with traumatic instability is important when planning treatment. Open or arthroscopic soft tissue repairs are usually sufficient when less than 25% of the width of the glenoid bone has been lost. Bone replacement techniques may be necessary when glenoid bone loss is greater than 25% of the glenoid width. Glenoid bone restoration techniques include the use of a tricortical iliac crest graft or the transfer of the coracoid process to the area of glenoid deficiency. Bone grafting becomes a strong consideration when soft tissue repairs have failed to restore stability. Treatment of these severe defects may be followed by osteoarthritis. The destabilizing effects of anterior glenoid bone defects are compounded by concurrent defects of the posterior lateral humeral head, commonly known as Hill-Sachs lesions, which can engage the glenoid defect. Large humeral head defects can be treated by transhumeral bone grafting techniques or osteoarticular allograft reconstruction. Prosthetic replacement of the proximal humerus is considered for humeral head defects involving more than 40% of the articular surface. Understanding the importance of humeral and glenoid bone deficiencies may help guide the treatment of recurrent anterior glenohumeral instability. PMID- 19218057 TI - Functional outcome of salvage of residual and recurrent deformities of clubfoot with Ilizarov technique. AB - Relapse may occur in severe clubfeet deformities whether treated surgically or nonsurgically. In this study, we evaluate the results of correction of residual and recurrent congenital clubfoot using soft tissue distraction or osteotomy using the Ilizarov external fixation system. This study included 35 feet in 28 patients who were treated between 1999 and 2007. Of these 16 feet in 13 patients with an average age of 13.7 years (range from 11 to 29 years) were treated by percutaneous calcaneal V-osteotomy and gradual correction by the Ilizarov method. Nineteen feet in 15 patients with an average age 10.5 years (range from 4 to 22 years) were treated by soft tissue distraction by the Ilizarov technique. The mean average follow up period was 5.6 years (range from 1 to 8 years). At the time of fixator removal, a plantigrade foot was achieved in 30 feet. Mild residual varus and equinus deformities were present in five feet. At the final follow foot pressure measurement showed recurrent or residual deformity in 10 feet (7 treated by osteotomy and 3 treated by soft tissue distraction). Recurrence may occur with both techniques, depending on many factors such as bone morphology, the number of the pervious operations and the degree of stiffness of the foot prior to the operation. PMID- 19218058 TI - The transverse forefoot arch demonstrated by a novel X-ray projection. AB - OBJECTIVE: Metatarsalgia is often treated by metatarsal osteotomy. Exact knowledge of the normal anatomy of the forefoot is essential for pre-operative planning. The objective of this study was to investigate the forefoot arch during maximal loading in a randomly selected population sample. METHODS: Two hundred subjects randomly selected from a municipality representative of Denmark were invited to interview and forefoot X-ray examination, including a novel horizontal X-ray projection by which the height of each metatarsal from the floor can be measured under maximal loading. RESULTS: One hundred and thirty-four subjects (79%) presented themselves for interview and X-ray examination. The study group was representative of the randomly selected population sample in terms of age, sex and incidence of metatarsalgia. The study verified that the interrelated geometry of the metatarsal heads in the AP plane corresponds to a parabola as suggested previously (Le Lievre's parabola). Also in the horizontal plane, the metatarsal heads generally form an arch, the transverse forefoot arch (TFA). Mean height was 3.91mm (S.E.=0.10). The individual height of the TFA varied from -1 to 10mm and was dependent on the width of the forefoot. The relative height of the arch (arch height divided by forefoot width) was independent of age and sex. A non-significant tendency towards a lower arch among subjects with metatarsalgia was observed. CONCLUSION: This population study demonstrated that the metatarsal heads constitute arches in both planes (Le Lievre's parabola in the AP plane and the transverse forefoot arch in the horizontal plane). This knowledge is essential for pre-operative planning in metatarsal osteotomy for metatarsalgia. Formulae for calculating the individual location of each metatarsal head were obtained. PMID- 19218059 TI - Chronic isolated distal tibiofibular syndesmotic disruption: diagnosis and management. AB - BACKGROUND: The current study outlines the pathology and treatment of persistent problems following isolated distal tibiofibular syndesmotic injuries. METHODS: A retrospective study was conducted to review patients with isolated chronic syndesmotic disruption who were managed in the authors' institute during 4 years, from January 2001 to January 2005. Patients with concomitant bony injuries or lateral ankle instability were excluded. RESULTS: The study included eleven patients with isolated syndesmotic disruption. All were males, with average duration of symptoms 4.7 years. Reconstruction of syndesmosis was achieved by semitendinosus tendon in all patients. The average hospital stay was 3.6 days, and the average follow up period was 3.1 years. According to West Point Ankle Score system, the average score after treatment was 95.4. CONCLUSION: Chronic isolated tibiofibular syndesmotic disruption appears amenable to accurate diagnosis and delayed stabilization. Arthroscopic management of the associated intraarticular pathology followed by reconstruction of torn syndesmosis can offer an excellent outcome. PMID- 19218060 TI - Some words about lower limb torsion and rotation problems. AB - The notions of torsion and rotation of the lower limb, loading and the progression of segmentary torsions are defined. It is shown that, while proximal dysmorphisms (anteversion of the hip) may lead to distal dysmorphisms, the latter, such as cavovalgus foot, may be implicated in the progression of dysmorphisms of both the knee (femoropatellar impingement syndromes) and the hip (residual anteversion). PMID- 19218061 TI - Effects of human amniotic fluid and membrane in the treatment of Achilles tendon ruptures in locally corticosteroid-induced Achilles tendinosis: an experimental study on rats. AB - BACKGROUND: To determine the effects of human amniotic fluid and membrane in the treatment of Achilles tendon ruptures, 72 tendons of 36 Wistar rats were injected with betamethasone sodium phosphate. METHODS: By the end of fourth week, both tendons were tenotomized and repaired, then the samples were divided into three groups. The first group was left untreated after suturing. Human amniotic fluid was injected to the second and amniotic fluid and membrane were both administered to the third group. Twenty-four tendons were scored at the end of the first week, and 24 at the end of the second week histopathologically, and 24 biomechanically at the end of the third week. RESULTS: There was a significant statistical difference only between the histopathological results of Groups 2 and 3 at the first week. CONCLUSIONS: Human amniotic membrane and fluid do not add anything to the healing process of Achilles tendon ruptures in the early phase. PMID- 19218063 TI - Tibial chondroblastic osteosarcoma--case report. AB - Osteosarcoma is a bone tumour that can occur in any bone. It most commonly occurs in the long bones of the extremities near the metaphyseal growth plates. The most common sites are femur (42%), tibia (19%), and humerus (10%). Other locations include the skull or jaw (8%) and pelvis (8%). A number of variants of osteosarcoma include conventional types (i.e., osteoblastic, chondroblastic, fibroblastic types) and telangiectatic, multifocal, parosteal, and periosteal types. We present a 10-year-old boy with Tibial Chondroblastic Osteosarcoma. The clinical features, diagnosis and management are discussed. PMID- 19218062 TI - Distal tibial giant cell tumour treated with curettage and stabilisation with an Ilizarov frame. AB - A 36-year-old man presented with a long history of pain in his left ankle. Imaging and biopsy confirmed this to be a giant cell tumour of bone. This was treated with curettage of the lesion and stabilisation with an Ilizarov frame. After a prolonged course of rehabilitation he has excellent function and has returned to hill walking and there is no evidence of recurrence of the giant cell tumour. PMID- 19218064 TI - Post-traumatic talectomy: a 60-year follow-up. AB - The long-term outcome of primary talectomy for trauma without tibiocalcaneal fusion has not been reported in the literature. We report a case of a primary talectomy for trauma in a 10-year-old patient with 60-year follow-up. The patient had minimal symptoms during this period which suggests that this procedure can have the desired effect in providing good long-term quality of life. PMID- 19218065 TI - A wear debris cyst following S.T.A.R. Total Ankle Replacement--surgical management. AB - We present a case of a 65-year-old gentleman who underwent a Total Ankle Replacement in January 2002. Four years later he required a revision procedure after developing edge loading and a polymer wear cyst within the fibula, which communicated directly with the ankle joint. PMID- 19218066 TI - Traumatic rupture of the tibialis anterior tendon associated with chronic tibialis posterior dysfunction. AB - Isolated rupture of the tibialis anterior and tibialis posterior tendon is uncommon with approximately 25 cases reported in the English literature. Rupture of the tibialis anterior in the presence of chronic tibialis posterior dysfunction has not been reported to date. We present a patient with a closed traumatic rupture of the tibialis anterior tendon which occurred on a background of a pre-existing tibialis posterior dysfunction which was being treated non operatively and discuss the successful operative management that was performed to reconstruct both tendons. PMID- 19218068 TI - Case report 'traumatic rupture of the tibialis anterior tendon associated with chronic tibialis posterior dysfunction' by George et al. PMID- 19218069 TI - A new technique for hammertoe arthrodesis. PMID- 19218070 TI - Bring back the biopsychosocial model for neck pain disorders. PMID- 19218071 TI - Medline success complements research dissemination and reach. PMID- 19218073 TI - Strategies used by physical therapists in the U.S. for treatment and prevention of the female athlete triad. AB - OBJECTIVE: To describe current methods of treatment and prevention used by physical therapists in the United States (US) for the female athlete triad. DESIGN: Descriptive analysis. SETTING: Physical therapists currently practicing in the U.S. PARTICIPANTS: 500 American Physical Therapy Association members. MAIN OUTCOME MEASURES: A survey was used to assess methods of practice for detecting, treating, and preventing the female athlete triad. Descriptive statistics summarized demographics about the survey participants. Likert scales and narrative descriptors determined the likelihood and frequency of employing certain treatment and prevention methods. RESULTS: Participants included 205 physical therapists for a 41% (205/500) response rate. Twenty-six percent (54/205) of the respondents used specific treatment methods, including education, for the female athlete triad; 48% of these respondents (26/54) incorporated preventative strategies other than screening; 13/54 (24%) assisted in athletic screening for the triad disorders. Physical therapists are more likely to talk to the athlete, the athlete's parents, and physicians when triad symptoms are suspected. The frequencies in which specific treatment and prevention strategies are utilized have been described. CONCLUSIONS: Physical therapists must be responsible for recognizing, treating and preventing the female athlete triad. This study emphasizes that there is a greater need for knowledge regarding the triad to be incorporated into physical therapy curriculums, continuing education programs and professional practice. PMID- 19218074 TI - Does the use of orthoses improve self-reported pain and function measures in patients with plantar fasciitis? A meta-analysis. AB - OBJECTIVES: To perform a meta-analysis examining the effects of foot orthoses on self-reported pain and function in patients with plantar fasciitis. DATA SOURCES: MEDLINE, SPORTDiscus, and CINAHL were searched from their inception until December 2007 using the terms "foot", "plantar fascia", "arch", "orthotic", "orthoses" and "plantar fasciitis". STUDY SELECTION: Original research studies which met these criteria were included: (1) randomised controlled trials or prospective cohort designs, (2) the patients had to be suffering from plantar fasciitis at the time of recruitment, (3) evaluated the efficacy of foot orthoses with self-reported pain and/or function, (4) means, standard deviations, and sample size of each group had to be reported. RESULTS: We utilised the Roos, Engstrom, and Soderberg (Roos, E., Engstrom, M., & Soderberg, B. (2006). Foot orthoses for the treatment of plantar fasciitis. Foot and Ankle International, 8, 606-611) night splint condition to compare our pooled orthoses results. The meta analysis results showed significant reductions in pain after orthotic intervention. The Roos et al.' (Roos, E., Engstrom, M., & Soderberg, B. (2006). Foot orthoses for the treatment of plantar fasciitis. Foot and Ankle International, 8, 606-611) study also showed significant reduction in pain after night splint treatment. The meta-analysis results also showed significant increases in function after orthotic use. In contrast, the Roos et al.' (Roos, E., Engstrom, M., & Soderberg, B. (2006). Foot orthoses for the treatment of plantar fasciitis. Foot and Ankle International, 8, 606-611) study did not show a significant increase in function after night splinting for 12 weeks. CONCLUSION: The use of foot orthoses in patients with plantar fasciitis appears to be associated with reduced pain and increased function. PMID- 19218075 TI - The effect of soft tissue release on delayed onset muscle soreness: a pilot study. AB - OBJECTIVES: To examine soft tissue release (STR) as an intervention for delayed onset muscle soreness (DOMS). DESIGN: A mixed-subjects experimental design was used. Participants performed 4 x 20 eccentric elbow extensions at 80% of 1RM. Participants received either STR (50%) or no treatment (50%). DOMS measurements were taken before the elbow extensions and at 0, 24, and 48 h afterwards. SETTING: The study was conducted at the University of Essex exercise physiology laboratory. PARTICIPANTS: Twenty male participants, unaccustomed to strength conditioning, completed the study. MAIN OUTCOME MEASURES: DOMS was evaluated using relaxed joint angle (RJA), active range of motion (AROM), passive range of motion (PROM), and arm girth measurements. Soreness ratings were measured using a 100 mm visual analogue scale (VAS). RESULTS: In both conditions there were post DOMS task increases in VAS ratings (p < 0.0001) and arm girths (p < 0.0001), and decreases in RJA (p < 0.0001), AROM (p < 0.0001), and PROM (p < 0.0001). STR group VAS scores were higher immediately (p < 0.01) and 48 h after treatment (p < 0.005). There were no other between-group differences and none of the measurements returned to baseline levels by 48 h. CONCLUSIONS: STR exacerbates the DOMS sensation yet does not seem to improve the rate of recovery during the first 48 h. PMID- 19218076 TI - Comparison of hip joint range of motion in professional youth and senior team footballers with age-matched controls: an indication of early degenerative change? AB - OBJECTIVES: To determine if there is evidence of abnormal hip joint range of motion (ROM) in youth and senior team professional footballers compared with matched controls. DESIGN: A case control study design was used. PARTICIPANTS: 40 professional footballers (20 youth and 20 senior team) and 40 matched control subjects. MAIN OUTCOME MEASURES: Bilateral measurements of passive hip internal rotation (IR), external rotation (ER), flexion, abduction and extension were made together with Faber's test and the hip quadrant. RESULTS: Youth and senior footballers had significantly less IR and Faber's range and significantly higher abduction than their respective controls (p < 0.001). Senior footballers also had significantly reduced IR (p < 0.05) and Faber's (p < 0.001) than the youth team. A higher proportion of senior footballers had positive hip quadrants (45% of all hips) compared to all other groups. No significant difference in hip ROM was found between dominant and non-dominant legs. CONCLUSIONS: A specific pattern of hip ROM was found in professional footballers which appeared to be different from controls. These changes may demonstrate the early stages of hip degeneration to which it has been shown ex-professional players are prone to. Hip joint ROM exercises may be necessary in these players to restore normal movement and prevent the onset of hip osteoarthritis (OA). PMID- 19218077 TI - Reliable lower limb musculoskeletal profiling using easily operated, portable equipment. AB - OBJECTIVES: Jumping performance and movement-based screening tests are commonly performed on athletes pre-season. Little is known regarding the test-retest reliability of performance tests and associated movement patterns using easily operated, portable and cost effective equipment. The purpose of this study was to establish the test-retest reliability of performance and video-derived movement characteristics of six screening tests measured simultaneously using a contact mat and two-dimensional video analysis. METHODS: Twenty-four subjects (17 male/7 female) were tested on three occasions 5-12 days apart to establish test-retest reliability of the measurement tools used. The tests used were anterior step-up, double and single-leg countermovement jumps, double and single-leg reactive (drop) jumps, and single-leg side springs. RESULTS: Measures of performance demonstrated less variability (mean-typical error expressed as a coefficient of variation, CV 2.8-4.3%) and higher reliability (intraclass correlation coefficients, ICC 0.98-0.99) for floor-based tests (preceding values) than drop tests (CV 10.3-13.2%, ICC 0.83-0.85). All video-derived movement characteristics demonstrated low variability (CV 3.2-4.8%), but lower rank order repeatability (ICC 0.43-0.79). CONCLUSION: The findings suggest that screening tests performed from the floor and measured simultaneously using a contact mat and two dimensional movement analysis are reliable, and trialling these tests for the pre season screening of athletes is supported. PMID- 19218078 TI - Minimal detectable change. PMID- 19218080 TI - Shigella targets the mitochondrial checkpoint of programmed necrosis. AB - In nonmyeloid cells, Shigella hijacks the mitochondrial checkpoint of cell death, thereby inducing a regulated form of necrosis depending on Bnip3 and cyclophilin D. Carneiro et al. (2009) describe the interplay between this program and a prosurvival response transmitted via the Nod1/Rip2/NF-kappaB/Bcl-2 axis, which determines the fate of infected cells. PMID- 19218081 TI - Mammalian PGRPs in the spotlight. AB - Peptidoglycan-recognition proteins (PGRPs) play a central role in the insect innate immune response to bacteria. In this issue of Cell Host & Microbe, Saha et al. (2009) report that the mammalian PGRP, PGLYRP-2, functions as a cytokine-like molecule in a PG-induced arthritis model. PMID- 19218082 TI - A mark of silence in malaria parasites. AB - Gene families in malaria parasites that encode hypervariable surface antigens display mutually exclusive expression that is controlled epigenetically. In this issue, Lopez-Rubio et al. (2009) show that the histone mark H3K9me3 is specifically associated with regulation of expression of these gene families within repressive centers located at the nuclear periphery. PMID- 19218083 TI - Aligning antimicrobial drug discovery with complex and redundant host-pathogen interactions. AB - Drug-resistant microorganisms pose an enormous threat to public health. Here we provide examples of experimental approaches that offer novel ways to think about drug development considering the complexity inherent to host-pathogen interactions. Emergent themes include (1) targeting pathogenicity rather than microbial growth, (2) targeting the host or host-pathogen interface rather than the pathogen, (3) facilitating pathogen-specific immune responses, and (4) utilizing systems-based approaches to identify new drug targets and validate drug efficacy. We posit that together these approaches may allow identification of drugs that disrupt pathogenesis and allow the immune system time to protect, but do not easily engender resistance. PMID- 19218084 TI - Shigella induces mitochondrial dysfunction and cell death in nonmyleoid cells. AB - Shigella rapidly kills myeloid cells via a caspase-1 inflammasome-dependent cell death mechanism. However, despite a critical role for nonmyeloid cells in the physiopathology of Shigella infection, the mechanism by which Shigella kills nonmyeloid cells remains uncharacterized. Here we demonstrate that, in nonmyeloid cells, Shigella infection induces loss of mitochondrial inner membrane potential, mitochondrial damage, and necrotic cell death through a pathway dependent on Bnip3 and cyclophilin D, two molecules implicated in the host oxidative stress responses. This mitochondrial cell death mechanism was potently counterbalanced by a Nod1-dependent Rip2/IKKbeta/NF-kappaB signaling pathway activated by the pathogen in the first hours of infection. Our results suggest that in nonmyeloid cells, oxidative stress pathways and signaling triggered by an intracellular bacterial pathogen are tightly linked and demonstrate the existence of specific Shigella-induced prodeath and prosurvival pathways converging at the mitochondria to control a necrotic cell death program. PMID- 19218085 TI - PGLYRP-2 and Nod2 are both required for peptidoglycan-induced arthritis and local inflammation. AB - Peptidoglycan recognition proteins (PGRPs) are structurally conserved from insects to mammals. Insect PGRPs have diverse host-defense functions. Mammalian PGRPs PGLYRP-1, PGLYRP-3, and PGLYRP-4 have bactericidal activity, while PGLYRP-2 has amidase activity. To extend the known functions of mammalian PGRPs, we examined whether they have immunomodulating activities in peptidoglycan-induced arthritis in mice. We demonstrate that PGLYRP-2 and Nod2 are both required for arthritis in this model. The sequence of events in peptidoglycan-induced arthritis is activation of Nod2, local expression of PGLYRP-2, chemokine production, and recruitment of neutrophils into the limbs, which induces acute arthritis. Only PGLYRP-2 among the four mammalian PGRPs displays this proinflammatory function, and PGLYRP-1 is anti-inflammatory. Toll-like receptor 4 (TLR4) and MyD88 are required for maturation of neutrophils before peptidoglycan challenge. Our results demonstrate that PGRPs, Nod2, and TLR4, representing three different types of pattern-recognition molecules, play interdependent in vivo roles in local inflammation. PMID- 19218086 TI - An intact cuticle in distal tissues is essential for the induction of systemic acquired resistance in plants. AB - Systemic acquired resistance (SAR), initiated by a plant upon recognition of microbial effectors, involves generation of a mobile signal at the primary infection site, which translocates to and activates defense responses in distal tissues via unknown mechanism(s). We find that an acyl carrier protein, ACP4, is required to perceive the mobile SAR signal in distal tissues of Arabidopsis. Although acp4 plants generated the mobile signal, they failed to induce the systemic immunity response. Defective SAR in acp4 plants was not due to impairment in salicylic acid (SA)-, methyl SA-, or jasmonic acid-mediated plant hormone signaling pathways but was associated with the impaired cuticle of acp4 leaves. Other cuticle-impairing genetic mutations or physical removal of the cuticle also compromised SAR. This cuticular requirement was relevant only during mobile signal generation and its translocation to distal tissues. Collectively, these data suggest an active role for the plant cuticle in SAR-related molecular signaling. PMID- 19218087 TI - Conserved herpesviral kinase promotes viral persistence by inhibiting the IRF-3 mediated type I interferon response. AB - A conserved herpesviral kinase, designated ORF36 in murine gamma-herpesvirus 68 (MHV-68), plays multiple vital roles in the viral life cycle. Here, we show by screening mutant viruses that ORF36 counteracts the antiviral type I interferon (IFN) response. ORF36 specifically binds to the activated form of interferon regulatory factor 3 (IRF-3) in the nucleus, inhibiting IRF-3 interaction with the cotranscriptional activator CBP and thereby suppressing the recruitment of RNA polymerase II to the interferon beta promoter. The anti-IFN function of ORF36 is conserved among herpesvirus subfamilies, although the conserved kinase activity is not absolutely required for this function. MHV-68 lacking ORF36 induces a greater interferon response and is attenuated in vitro and in vivo, where acute viral infection in the lung and latency in the spleen are compromised. Our data suggest that herpesviruses have evolved within their conserved kinase an anti-IFN activity critical for evasion of host immunity and for persistence. PMID- 19218088 TI - Genome-wide analysis of heterochromatin associates clonally variant gene regulation with perinuclear repressive centers in malaria parasites. AB - Clonally variant gene families underlie phenotypic plasticity in Plasmodium falciparum, a process indispensable for survival of the pathogen in its human host. Differential transcription of one of these gene families in clonal parasite lineages has been associated with chromatin modifications. Here, we determine the genome-wide distribution in P. falciparum of a histone mark of heterochromatin, trimethylation of histone H3 lysine 9 (H3K9me3), using high-resolution ChIP-chip analysis. We show that H3K9me3 is specifically associated with clonally variant gene families, which are clustered on subtelomeric and some chromosome internal regions. High levels of H3K9me3 correlate with genes localized to the nuclear periphery, implying chromosome loop formation. Disruption of the histone deacetylase PfSir2 causes changes in H3K9me3 that are discontinuous along chromosomes and associated with disrupted monoallelic transcription. Our data point to the existence of perinuclear repressive centers associated with control of expression of malaria parasite genes involved in phenotypic variation and pathogenesis. PMID- 19218089 TI - Host-parasite interactions revealed by Plasmodium falciparum metabolomics. AB - Intracellular pathogens have devised mechanisms to exploit their host cells to ensure their survival and replication. The malaria parasite Plasmodium falciparum relies on an exchange of metabolites with the host for proliferation. Here we describe a mass spectrometry-based metabolomic analysis of the parasite throughout its 48 hr intraerythrocytic developmental cycle. Our results reveal a general modulation of metabolite levels by the parasite, with numerous metabolites varying in phase with the developmental cycle. Others differed from uninfected cells irrespective of the developmental stage. Among these was extracellular arginine, which was specifically converted to ornithine by the parasite. To identify the biochemical basis for this effect, we disrupted the plasmodium arginase gene in the rodent malaria model P. berghei. These parasites were viable but did not convert arginine to ornithine. Our results suggest that systemic arginine depletion by the parasite may be a factor in human malarial hypoargininemia associated with cerebral malaria pathogenesis. PMID- 19218090 TI - Drosophila intestinal response to bacterial infection: activation of host defense and stem cell proliferation. AB - Although Drosophila systemic immunity is extensively studied, little is known about the fly's intestine-specific responses to bacterial infection. Global gene expression analysis of Drosophila intestinal tissue to oral infection with the Gram-negative bacterium Erwinia carotovora revealed that immune responses in the gut are regulated by the Imd and JAK-STAT pathways, but not the Toll pathway. Ingestion of bacteria had a dramatic impact on the physiology of the gut that included modulation of stress response and increased stem cell proliferation and epithelial renewal. Our data suggest that gut homeostasis is maintained through a balance between cell damage due to the collateral effects of bacteria killing and epithelial repair by stem cell division. The Drosophila gut provides a powerful model to study the integration of stress and immunity with pathways associated with stem cell control, and this study should prove to be a useful resource for such further studies. PMID- 19218091 TI - Mind-body interactions in pain: the neurophysiology of anxious and catastrophic pain-related thoughts. AB - The well-accepted biopsychosocial model proposes that the experience of pain and responses to it result from a complex interaction of biological, psychological, and social factors. However, the separation of these constructs is substantially artificial, and we presume that psychological processes have biological effects, that biological processes affect an individual's psychosocial environment, and so on. Considerable research has demonstrated that pain-coping strategies influence perceived pain intensity and physical functioning, and individual differences in styles of pain coping even shape the persistence of long-term pain complaints in some populations. A good deal of this coping research has focused on catastrophizing, which is a generally maladaptive cognitive and emotional mental set that involves feelings of helplessness when in pain, rumination about pain symptoms, and magnification of pain-related complaints. Collectively, catastrophizing has been consistently associated with heightened experiences of pain across a variety of samples. Although catastrophic thinking regarding pain related symptoms is often classified under the "psychologic" category within the broader biopsychosocial model, we propose that catastrophizing exerts biologic effects that may account for some of its negative consequences. In general, the cognitive and affective processes captured within the construct of catastrophizing may exert effects on the neuromuscular, cardiovascular, immune, and neuroendocrine systems, and on the activity in the pain neuromatrix within the brain. The interface between pain-related neurobiology and processes such as pain-related catastrophizing represents an important avenue for future pain research. PMID- 19218093 TI - Identification of deletion carriers in hemophilia B: quantitative real-time polymerase chain reaction or multiple ligation probe amplification. AB - Gross deletions in the F9 gene are easily detected by routinely sequencing hemophilia B-affected men. Nevertheless, a carrier diagnosis proves difficult as the presence of a normal allele does not recognize the partial or complete loss of the F9 gene and may be challenging if no DNA sample from affected men is available. This work aimed to identify hemophilia carriers in 2 families in which gross deletions of the F9 gene could be expected. The indirect genetic study was not conclusive, and sequencing did not show genetic defects in family 1. A real time polymerase chain reaction (RT-PCR) assay using SYBR Green revealed the deletion of a copy of exon 8 in 3 women, whereas the multiple ligation-dependent probe amplification (MLPA) assay showed the deletion of a copy of exons 7 and 8 in these 3 women. These studies enabled us not only to rule out a pregnant woman as a carrier but also to confirm a complete deletion of the gene in the patient from family 2 and the heterozygous state of his mother. The advantages that the MLPA method offers are the identification of a multiple exon deletion in the same assay and commonly used technology. The RT-PCR technology used involves standardizing and analyzing each exon independently. PMID- 19218094 TI - Sulodexide suppresses inflammation in human endothelial cells and prevents glucose cytotoxicity. AB - Sulodexide is a mixture of heparin and dermatan sulfate with antithrombotic and profibrynolytic activity. Individual reports suggest the anti-inflammatory action of sulodexin. The goal of this study was to evaluate the effect of sulodexide on the release of the inflammatory mediators from endothelium in normal conditions and in cells chronically exposed to glucose. The experiments were performed on in vitro cultured human umbilical endothelial cells kept for 7 days in standard medium or in the same medium but supplemented with glucose 30 mmol/L. Sulodexide was added to the culture medium in concentrations of 0.125 lipase releasing unit (LRU)/mL, 0.25 LRU/mL, and 0.5 LRU/mL Spontaneous generation of oxygen-derived free radicals and the release of monocyte chemotactic protein-1 (MCP-1) and interleukin-6 (IL-6) from the studied cells was evaluated. Additionally, the healing of the injured mesothelium was studied in the presence of sulodexide and glucose. Sulodexide caused the inhibition of the intracellular generation of free radicals in a dose-dependent manner (maximally by 32%, P < 0.01), as well as the inhibition of MCP-1 (maximally by 60%, P < 0.001) and IL-6 (maximally by 69%, P < 0.01). Cells cultured in a medium with glucose 30 mmol/L generated more free radicals (+20%, P < 0.05) and released more MCP-1 (+113%, P < 0.001) and IL-6 (+26%, P < 0.05). Cell monolayers treated with glucose had a decreased ability to heal after mechanical injury (-28%, P < 0.001). All these glucose effects were reversed when cells were exposed to sulodexide simultaneously. The results of our study demonstrate a significant anti-inflammatory action of sulodexide in the endothelial cells and a protective effect of that drug against glucose cytotoxicity. PMID- 19218092 TI - Renin-angiotensin-aldosterone system-mediated redox effects in chronic kidney disease. AB - The renin-angiotensin-aldosterone system (RAAS) is central to the pathogenesis of hypertension, cardiovascular disease, and kidney disease. Evidence supports various pathways through which a local renal RAAS can affect kidney function, hypertension, and cardiovascular disease. A prominent mechanism seems to be the loss of reduction-oxidation (redox) homeostasis and the formation of excessive free radicals. Free radicals such as reactive oxygen species (ROS) are necessary in normal physiologic processes, which include the development of nephrons, erythropoeisis, and tubular sodium transport. However, the loss of redox homeostasis contributes to proinflammatory and profibrotic pathways in the kidney that in turn lead to decreased vascular compliance, podocyte pathology, and proteinuria. Both the blockade of the RAAS and the oxidative stress produce salutary effects on hypertension and glomerular filtration barrier injury. Thus, the focus of current research is on understanding the pathophysiology of chronic kidney disease in the context of an increased RAAS and unbalanced redox mechanisms. PMID- 19218095 TI - Regulation of thrombin generation by TFPI in plasma without and with heparin. AB - The purpose of this study was to investigate the impact of recombinant glycosylated TFPI (rg-TFPI) from BHK cells, nonglycosylated TFPI (r-TFPI) from Escherichia coli, and truncated TFPI (1-161) on thrombin generation (TG) in plasma treated with and without heparin in vitro and ex vivo. Fasting plasma samples were collected from 6 healthy persons. TG was assessed by the calibrated automated thrombography (CAT) method. The addition of increasing concentrations (0-200 ng/mL) of different TFPI caused a 5% to 30% prolongation of lag time for TF (3.0 pM) induced TG, with the most pronounced effect for rg-TFPI and the least pronounced effect for truncated TFPI, but without affecting endogenous thrombin potential (ETP) in TF-induced coagulation. Removal of native TFPI from plasma by anti-TFPI IgG treatment shortened lag time by 35 +/- 4% without affecting ETP. Increasing concentrations (0-200 ng/mL) of various TFPI in the presence of low heparin concentrations (0.1 IU/mL) prolonged lag time and decreased ETP by 25% to 75% with the most prominent effect promoted by glycosylated full-length TFPI. The effect of neutralizing antibodies against TFPI and antithrombin (AT) was studied in plasma in the presence of heparin administered in vitro or ex vivo. The results revealed that TFPI and AT acted in synergy as inhibitors of coagulation in terms of the effect on both initiation (lag time) and propagation (ETP). Our data demonstrated that the CAT assay appropriately assessed the impact of TFPI on initiation and propagation of TG in a physiological plasma milieu with and without heparin. TFPI contributed significantly to regulation of coagulation initiation (lag time). The C-terminal region and, to a lesser extent, glycosylation of the TFPI molecule were essential for its anticoagulant function in the absence and presence of heparin. PMID- 19218096 TI - Acteylcholinesterase inhibitor rivastigmine enhances cellular defenses in neuronal and macrophage-like cell lines. AB - Neuroprotection mediated by the cellular heat shock response offers 1 clinical strategy to prevent, stabilize, and possibly reverse neurodegenerative processes. Although damaged proteins are thought to be the primary stimulus for the heat shock response, several studies indicate that pharmaceutical agents can either directly induce the heat shock transcription factor (Hsf1) or enhance its activation during different forms of cellular stress. Because Hsf1 is now known to combat the proteotoxicity of aging and has a central role in modulating amyloid aggregation, pharmacologic interventions to strengthen Hsf1 action may have important implications for preventing neurodegeneration linked to altered and damaged proteins such as observed in Alzheimer's disease. Given reports that some agents for the treatment of Alzheimer's disease have neuroprotective properties, this project investigated whether rivastigmine, which is an acetyl and butaryl cholinesterase inhibitor, mediates the neuroprotection of the neuronal-like cell line SH-SY5Y. The cells were exposed to various concentrations of rivastigmine to determine whether the drug protected cells from toxic injury and induced the 1st phase of the cellular heat shock response. In all, 100 micromol/L rivastigmine decreases cell death by 40% compared with untreated cells. This concentration enhances Hsf1 activation by strengthening both its multimerization and its phosphorylation, which leads to increased messenger RNA (mRNA) for hsp70. Therefore, one of the putative neuroprotective mechanisms of rivastigmine seems to be mediated through the heat shock response. These results also are observed in cultured macrophage-like cells, which suggests a future clinical tool for monitoring pharmacologically improved stress responses in peripheral blood mononuclear cells during treatment of Alzheimer disease. PMID- 19218097 TI - [A double suicide gene system driven by KDR promoter selectively kills human hepatic carcinoma cells and human umbilical vein endothelial cells]. AB - OBJECTIVE: To study the selective killing effects of adenovirus (Ad)-mediated double suicide gene system driven by KDR promoter (KDR-CdglyTK) on the human hepatic carcinoma cells and human umbilical vein endothelial cells (HUVECs). METHODS: KDR-expressing BEL-7402 and HUVECs and HepG2 cells that did not express KDR were infected by KDR-CdglyTK, and the infection efficiency and the expression of CdglyTK in the cells was detected by RT-PCR. The infected cells were treated with the the prodrugs 5-FC and GCV at different concentrations, and the cell killing effects and bystander effects were evaluated by MTT method. RESULTS: At the multiplicity of infection (MOI) of 100, the recombinant AdKDR-CDglyTK showed similar infection efficiency in the 3 cell lines. RT-PCR demonstrated CDglyTK expression in the recombinant adenovirus and the 3 infected cell lines. BEL-7402 and HUVECs infected by the KDR-CdglyTK, but not the HepG2 cells, were highly sensitive to the prodrugs (P<0.001). Bystander effects of the double suicide gene system were observed in the coculture of the infected and non-infected BEL-7402 and HUVECs. CONCLUSION: The double suicide gene system driven by KDR promoter has specific killing effect on KDR-expressing hepatocellular carcinoma cells and HUVECs. PMID- 19218098 TI - [Expression of interleukin-1beta and tumor necrosis factor-alpha in the synovium and synovial fluid of patients with Kashin-Beck disease and osteoarthritis]. AB - OBJECTIVE: To investigate the role of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) in the pathogenesis and progression of Kashin Beck disease (KBD) and primary osteoarthritis (OA). METHODS: The synovium and synovial fluid of the knee joint were collected from 20 adult patients with KBD, 18 with OA and 19 with meniscus injury (controls). The expression of IL-1beta and TNF-alpha in the synovium were analyzed by immunohistochemistry staining, and the levels of IL-1beta and TNF-alpha in the synovial fluid were determined by enzyme linked immunosorbent assay. RESULT: IL-1beta and TNF-alpha expressions in the synovium and their levels in the synovial fluid were significantly higher in patients with KBD and OA than in patients with meniscus injury (P<0.05), but comparable between KBD and OA groups (P>0.05). CONCLUSION: IL-1beta and TNF-alpha may play an important role in the pathogenesis and progression of KBD and OA. PMID- 19218099 TI - [Effects of A68930 on rhythmical respiratory discharge in isolated neonatal rat brainstem slice]. AB - OBJECTIVE: To investigate the role of dopamine-1 receptor in the modulation of basic respiration rhythm. METHODS: Newborn SD rat (0-3 days, n=20) brain stem slices containing the medial region of the nucleus retrofacialis (mNRF) were prepared with the hypoglossal nerve roots retained. The respiratory rhythmical discharge activity (RRDA) of the hypoglossal nerve was recorded using suction electrodes on these preparations, and the effects of dopamine-1 receptor on RRDA were investigated by application of the specific agonist of dopamine-1 receptor A68930 at different concentrations (0, 1, 2, and 5 micromol/L) in the perfusion solution. RESULTS: The respiratory cycles (RC) and the expiratory time (TE) decreased progressively with gradual increment of the integrated amplitude (IA) after A68930 administration, and their changes were the most conspicuous at 5 min after the administration. A68930 at the concentrations of 2 and 5 micromol/L resulted in the most obvious changes in RC, TE, and IA (P<0.05), but IA exhibited no significant variation at 1 min after perfusion with 2 micromol/L A68930 (P>0.05). RC and TE were gradually shortened after treatment with increasing concentrations of A68930, which also caused gradual increment of IA, and at the concentration of 5 micromol/L, RC, TE, and IA all showed the most obvious changes (P<0.01). CONCLUSIONS: Dopamine-1 receptor plays a role in the modulation of RRDA in isolated neonatal rat brainstem slice. A68930 may increase the frequency of respiration by shortening TE and enhance the respiratory activity by increasing the amplitude of inspiratory discharge of the respiratory neurons. PMID- 19218100 TI - [Binding activity of polypeptide containing human Na+, K+-ATPase alpha1 subunit M1-M2 extracellular segment]. AB - OBJECTIVE: To assess the binding activity of polypeptide containing human Na+, K+ ATPase alpha1 subunit M1-M2 extracellular segment (HES1 derivative). METHODS: HES1 derivative was synthesized by Fmoc method and purified by high-performance liquid chromatography-mass spectrometry, and its binding activity was identified by radioligand binding assay. RESULTS: 3H-ouabain and synthetic HES1 derivative showed some binding activity with the equilibrium dissociation constant (KD) of 24.58 nmol/L, with the the receptor density of 492.43 fmol x mg(-1) pro. and IC50 of 3.078 x 10(-7) mol/L. CONCLUSION: HES1 derivative can bind to ouabain and has the potential of becoming an effective therapeutic agent. PMID- 19218101 TI - [Value of virtual surgery in arterial reconstruction in liver recipients with type II hepatic artery variation]. AB - OBJECTIVE: To investigate the value of virtual surgery in hepatic artery reconstruction in liver recipients with type II hepatic artery variation. METHODS: A patient with cholangiocellular carcinoma and a healthy individual were scanned using 64-slice spiral CT, and image segmentation and three-dimensional (3D) reconstruction were performed using an image processing system. The 3D models in STL format were then imported to the FreeForm Modeling System for smoothing and refinement. Hepatic artery reconstruction was performed in simulated liver transplantation using the virtual surgery system with force feedback (PHANTOM). RESULTS: The reconstructed model contained the liver, hepatic arteries, biliary system, and bile duct tumor emboli and displayed the entire branching of the hepatic artery with type II variation. Using the virtual surgery system, arterial reconstruction was performed by anastomosing the donor celiac trunk and the recipient abdominal aorta with the virtual scalpel and needle. CONCLUSION: The reconstructed model allows clearer views of the 3D structures of the arteries in the liver and helps in preoperative preparations and surgical planning of artery reconstruction during liver transplantation. This approach may also help reduce the surgical risks and potential complications. PMID- 19218102 TI - [Effect of AT-1 alpha receptor gene silencing on nuclear factor-kappaB activity in hepatic Kupffer cells]. AB - OBJECTIVE: To investigate the effect of angiotensin II type-1 (AT-1) alpha receptor gene silencing on nuclear factor-kappaB (NF-kappaB) activity in hepatic Kupffer cells. METHODS: The expression of AT-1 alpha receptors in primary isolated cultured hepatic Kupffer cells was detected by immunohistochemistry. pSilencer/AT-1 alpha receptor siRNA plasmids were transfected into Kupffer cells, which were subsequently exposed to 10(-6) mol/L angiotensin II (Ang II) for 60 min. The changes in the DNA binding activity of NF-kappaB in the cells was assessed using electrophoretic gel mobility shift assay (EMSA). RESULTS: AT-1 alpha receptor expression was detected in Kupffer cells. NF-kappaB DNA binding activity was markedly increased in Kupffer cells after Ang II stimulation, and obviously inhibited by transfectiom with pSilencer/AT-1 alpha receptor siRNA plasmid. CONCLUSION: Ang II stimulation of Kupffer cell results in increased activation of NF-kappaB via AT-1 alpha receptor. PMID- 19218103 TI - [Ultrasonic study of deep-vein diameter and blood flow spectrum changes in full term pregnant women]. AB - OBJECTIVE: To explore the changes in lower limb deep vein diameters, blood flow velocity and blood biochemistry in full-term pregnant women for early diagnosis and treatment of prothrombotic state. METHODS: One hundred and twenty-eight full term pregnant women at high risk of thrombosis (Group A), 61 healthy full-term pregnant women (Group B), and 42 healthy non-pregnant women (Group C) underwent high-resolution color Doppler ultrasound (CDU) for examining the deep veins of the lower limbs. The hematological indexes such as D-D, PLT, HGB, HCT, TT, APTT, PT, and FbgC were also observed in these 3 groups. RESULTS: Compared to Group B, the women in group A showed significantly increased diameters of the common femoral veins (CFV) and left superficial femoral vein (SFV), HCT and DD, but with significantly decreased peak blood flow in the bilateral popliteal veins (POPV) (P<0.01) and increased left POPV diameter (P=0.034). Compared to those in group C, the diameters of the bilateral CFVs, SFVs, POPV, and posterior tibial veins (PTVs) were significantly increased, but the peak blood flow in the bilateral CFVs and POPVs were significantly reduced in groups A and B; the PLT, HGB, HCT, DD, TT, APTT, PT, and FbgC also showed significant changes in groups A and B (P<0.01). CONCLUSION: The full-term pregnant women are at higher risk of prothrombotic state than non-pregnant women, and the full-term pregnant women with the high risk factors for thrombosis are more likely to have prothrombotic state than healthy full-term pregnant women. CDU examination of the lower limb deep veins can be of value in the diagnosis of prothrombotic state. PMID- 19218104 TI - [Relationship between Tiam-1 expression and the biological behaviors of nasopharyngeal carcinoma]. AB - OBJECTIVE: To investigate the relationship between the protein expression of T lymphoma invasion and metastasis gene 1 (Tiam-1) and the biological behaviors of nasopharyngeal carcinoma (NPC). METHODS: Immunohistochemistry was performed to detect the expressions of Tiam-1 protein in 60 specimens of NPC tissue, 20 specimens of chronic nasopharyngitis (CN) tissue, and 6 tumor tissues from nude mice inoculated with metastatic human NPC cells. RESULTS: The positivity rate and average score for Tiam-1 expression were significantly higher in NPC tissues than in CN tissue (63.33% vs 36.67%, 2.9167 +/- 1.3057 vs 0.7000 +/- 0.9234; chi(2)=20.429, P=0.001; t=7.0162, P=0.0000, respectively). No difference was found in Tiam-1 expression among NPC patients in different T stages (F=2.36, P=0.0811), while the expression differed significantly between the patients with lymph node metastasis and those without metastasis, and also between patients with organ metastasis and those without (P=0.0001). High Tiam-1 expressions were found in the tumor tissues in nude mice inoculated with metastatic NPC cells. CONCLUSION: Tiam-1 expression is closely associated with the invasiveness and metastasis of NPC, indicating that Tiam-1 is an important factor that promotes the invasion and metastasis of NPC. PMID- 19218105 TI - [Hybrid interpolation for CT metal artifact reducing]. AB - Numerous interpolation-based methods have been described for reducing metal artifacts in CT images, but due to the limit of the interpolation methods, interpolation alone often fails to meet the clinical demands. In this paper, we describe the use of quartic polynomial interpolation in reconstruction of the images of the metal implant followed by linear interpolation to eliminate the streaks. The two interpolation methods are combined according to their given weights to achieve good results. PMID- 19218106 TI - [Proteinchip-flight mass spectrometry for screening differentially expressed proteins in the serum of cervical cancer patients]. AB - OBJECTIVE: To identify the differentially expressed proteins in the serum of patients with cervical cancer for use as the biomarkers for early diagnosis of cervical cancer. METHODS: Surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS) with weak cationic chips (CM10) was used to examine the serum samples of 24 patients with cervical squamous cell carcinoma and 25 age-matched healthy women. The protein fingerprints were obtained, and bioinformatic analysis was performed to identify the differentially expressed proteins in the serum of the patients. RESULTS: Fifty-two differentially expressed proteins were detected in the serum of cervical cancer patients (P<10( 5)), among which 6 proteins with mass/charge ratio of 4173.77, 5903.09, 6087.12, 10716.9, 6109.61 and 3397.41, respectively, showed lowered expression in the serum of cervical cancer patients. Two diagnostic models for cervical cancer were generated using software, including one consisting of the 4173.77(M/Z) protein with the diagnostic specificity of 96% and sensitivity of 75% for cervical cancer and the other consisting of 3 proteins at 5335.81(M/Z), 7562.99(M/Z), and 9287.89(M/Z) with specificity of 91.67% and sensitivity of 96.0%. CONCLUSION: Cervical cancer patients show different serum protein expression profile from healthy women. The 6 differential proteins identified may serve as specific serum biomarkers in close relation to the origin and progression of cervical cancer. PMID- 19218107 TI - [Association of the T1270533G polymorphism in GSTM1 gene coding region with susceptibility to nasopharyngeal carcinoma in a Chinese population]. AB - OBJECTIVE: To explore the association of T1270533G polymorphism in the glutathione S-transferase M1 (GSTM1) gene with the susceptibility to nasopharyngeal carcinoma (NPC) and clinical phenotype of NPC in Chinese population. METHDOS: The genomic DNAs were obtained from 27 Chinese subjects, and the single nucleotide polymorphism (SNP) in all the exons and relevant intron exon boundaries of GSTM1 were determined by PCR and direct sequencing. A case control study was performed to analyze the SNP site T1270533G (the rare allele frequency is 22.2% in Chinese population) in the coding region by means of tetra primer amplification refractory mutation system-polymerase chain reaction (ARMS PCR) and sequencing. RESULT: Sequence analysis identified 29 SNPs in GSTM1 gene, among which 13 SNPs presented high linkage disequilibrium with each other. No obvious relations were found between the variation in the coding region T1270533G and the clinical phenotype of NPC (RR=0.170, 95% CI =0.95-0.306 for TT homozygotes). CONCLUSION: The missense mutation in the coding region T1270533G of GSTM1 gene that causes an amino acid change does not affect the detoxification function of GSTM1, and the T1270533G polymorphism does not have apparent relations to NPC susceptibility in Chinese subjects in Guangdong Province. PMID- 19218108 TI - [In vitro cytotoxicity of 131I-Rituximab against B-cell lymphoma cells]. AB - OBJECTIVE: To study the specific cytotoxicity of (131)I-Rituximab against CD20 positive B-cell lymphoma cells. METHODS: Rituximab was labeled with (131)I using IODO-GEN method, and the dose-effects of various concentrations of (131)I Rituximab, (131)I alone and Rituximab in Raji cells were evaluated by MTT assay to determine the optimal dose according to the dose-effect curves. The cytotoxicity of (131)I-Rituximab, (131)I and Rituximab was assessed in CD20 positive Ramos (RA-1) cells, Raji cells and CD20-negative Molt-4 cells according to the changes of the survival rates. Giemsa staining was used to evaluate the antitumor effect of (131)I-Rituximab in Raji cells by measuring the mitosis index (MI). RESULTS: (131)I-Rituximab presented with a dose-dependent cytotoxicity against Raji cells. At the specific activity of 60 microCi/ml, (131)I-Rituximab resulted in significantly higher growth inhibition rate of the cells than Rituximab (P<0.05). The inhibition rate of Raji cells treated with (131)I Rituximab, (131)I, or Rituximab for 96 h were comparable with the rates in Ramos RA-1 cells, but significantly higher than the rates in Molt-4 cells. The MI values in (131)I-Rituximab group were significantly lower than those in the other groups (P<0.001). CONCLUSION: (131)I-Rituximab can induce specific cytotoxicity against CD20-positive tumor cells, and may potentially serve as an agent for targeted radioimmunotherapy for CD20-positive B-cell lymphoma. PMID- 19218109 TI - [Expression of heparin-binding epidermal growth factor-like growth factor in patients with gastric carcinoma]. AB - OBJECTIVE: To investigate the expression of heparin-binding epidermal growth factor-like growth factor (HB-EGF) in patients with gastric carcinoma in different stages. METHODS: The expressions of HB-EGF protein and mRNA in normal gastric tissues, metaplasic intestinal mucosa, early-stage gastric cancer and advanced-stage gastric cancer tissues were detected by immunohistochemistry and in situ hybridization. RESULTS: HB-EGF expression was only detected in the parietal cells of the gastric fundic glands and in gastrin cells of the pyloric glands in normal gastric tissues. Weak HB-EGF expression was detected in the epithelial cells of the gastric mucosa in intestinal metaplasic mucosa, and the expression increased in all layers of the gastric mucosa in early-stage gastric cancer. Intense HB-EGF expression was observed in advanced gastric cancer. CONCLUSION: Increased HB-EGF expression may be implicated in the pathogenesis and development of gastric carcinoma. PMID- 19218110 TI - [Expression of Aurora-B in human glioma tissue and its significance]. AB - OBJECTIVE: To study the expression of Aurora-B in human glioma tissue and its significance. METHODS: The total RNA was extracted from 41 human glioma tissues and 11 normal brain tissues by Trizol reagent. After reverse transcription of the total RNA into cDNAs, Aurora-B mRNA expressions in these samples were detected by quantitative real-time PCR. The protein expression in these samples was detected using immunohistochemical staining. RESULTS: Aurora-B mRNA and protein expressions were significantly increased in glioma tissues as compared with those in normal brain tissues. CONCLUSION: Aurora-B mRNA and protein show markedly higher expressions in glioma tissue, suggesting that Aurora-B may be one of the malignant biomarkers in the pathogenesis and progression of human glioma. PMID- 19218111 TI - [Effect of high glucose exposure on connective tissue growth factor expression in cultured human renal tubular epithelial cells and the role of p38MAPK pathway]. AB - OBJECTIVE: To observe the effect of high glucose exposure on connective tissue growth factor (CTGF) expression in cultured human renal tubular epithelial cells and investigate the role of p38MAPK pathway in this process. METHODS: Human renal tubular epithelial cells (HKC) with and without SB203580 pretreatment were cultured in the presence of high glucose levels for 24, 48, 72, 96 h and 20 days. RT-PCR, immunohistochemical staining, indirect fluorescence staining and Western blotting were used to detect the changes in CTGF mRNA and protein expressions in the cells after the treatment. RESULTS: Low levels of CTGF mRNA and protein were detected in cultured HKC cells, and after high glucose treatment, the mRNA expression increased gradually and reached the peak level at 48 h, then followed by gradual decrease till recovering the baseline level at 96 h. Prolonged high glucose treatment for 20 days resulted in persisted high CTGF mRNA expression twice the level in the control group. The expression level of CTGF protein also increased progressively as the treatment time then prolonged, and long-term (20 days) treatment increased the expression by 4 folds in comparison with the expression in the control cells. SB203580 significantly inhibited the increase in the expressions of CTGF mRNA and protein stimulated by high glucose treatment. CONCLUSION: High glucose treatment can increase CTGF mRNA and protein expressions in cultured human renal tubular epithelial cells, suggesting that increased CTGF levels is a key event in the pathogenesis of renal tubulo-interstitial fibrosis in patients with diabetic nephropathy. p38MAPK pathway may also participate in this process. PMID- 19218112 TI - [Epithelial sodium channel subunit mRNA expression in acutely isolated rat alveolar type II cells]. AB - OBJECTIVE: To investigate the epithelial sodium channel (ENaC) subunit mRNA expression in acutely isolated rat alveolar type II (ATII) cells. METHODS: Acutely isolated ATII cells from 20 SD rats were purified and ENaC alpha, beta, gamma-subunit mRNA levels were determined by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: mRNA expressions of all the subunits were detected in the ATII cells, and ENaC alpha-subunit mRNA showed significantly higher expression than beta- and gamma-subunit mRNAs, and the expressions of the latter two mRNAs were comparable. CONCLUSION: As the predominant ENaC subunit expressed at the mRNA level in rat ATII cells, the alpha-subunit of ENAC plays an important role in alveolar fluid clearance. PMID- 19218113 TI - [Risk factors of ventilator-associated pulmonary Candida infection and drug resistant analysis]. AB - OBJECTIVE: To investigate the risk factors for pulmonary Candida infection in association with mechanical ventilation and analyze the drug resistance profile of the Candida species that cause the infection. METHODS: A retrospective analysis was conducted 114 patients receiving mechanical ventilation for over 48 h. According to the presence of pulmonary Candida infections, these patients were divided into infected group (n=50, 43.9%) and non-infected group (64 cases). Univariate analysis and multivariate logistic regression analysis were performed to identify the risk factors for the infection, and drug sensitivity test was carried out to evaluate the drug resistance of the Candida species. RESULTS: Univariate analysis and multivariate logistic regression showed that the presence of at least two underlying diseases (OR=4.758, P=0.009), frequent changes of antibiotics (OR=6.128, P=0.001), and blood albumin below 25 g (OR=15.829, P=0.011) were the independent risk factors for pulmonary Candida infection associated with mechanical ventilation, and prophylactic antifungal treatment (OR=0.062, P=0.012) was a protective factor. Drug sensitivity test showed that Candida albicans was sensitive to most of the antifungal agents (100.0%), but the non-albicans Candida species were resistant to fluconazol (50.0%) and Itraconazole (38.5%). CONCLUSION: Poor general conditions and frequent changes of antibiotics are the major risk factors for pulmonary Candida infection in patients receiving mechanical ventilation. Drug resistant analysis is helpful in the treatment of the infections. PMID- 19218114 TI - [Effect of phosphorylated-ERK1/2 on inducible nitric oxide synthase expression in the substantia nigra of mice with MPTP-induced Parkinson disease]. AB - OBJECTIVE: To investigate the effect of phosphorylated-ERK1/2 (p-ERK1/2) on inducible nitric oxide synthase (iNOS) expression in the substantia nigra (SN) of a mouse model of Parkinson's disease (PD), and explore the possible mechanism of dopaminergic (DA) neuron loss in the SN of the midbrain in PD. METHODS: PD was induced by intraperitoneal injections of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine(MPTP) in C57BL/6N mice, and the behavioral changes of the PD mouse model were observed. Immunohistochemistry and Western blotting were used to detect the number of positive cells and the expressions of tyrosine hydroxylase (TH), p-ERK1/2 and iNOS in the SN of the PD mice, and their changes following Rg1 treatment were assessed. RESULTS: The PD mice exhibited typical symptoms of PD, in which the number of TH-positive neurons and TH expression were significantly reduced by about 77% and 75% (P<0.01), respectively, 7 days after the 5th injection of MPTP as compared with those in the control group. Rg1 pretreatment significantly decreased the number of TH-positive neurons and TH expression by 44% and 41% (P<0.01), respectively. p-ERK1/2 expression was not observed in the cell nuclei until 1.5 h after the third injection of MPTP, and increased markedly at 6 h. Rg1 pretreatment significantly inhibited the expression of p-ERK1/2 and iNOS (P<0.01). A significant positive correlation was noted between the expression of p-ERK1/2 and iNOS (P<0.01). CONCLUSION: P-ERK1/2 may regulate the expression of iNOS to induce DA neuron loss in the SN of PD, and Rg1 may protect the DA neurons possibly by depressing nuclear translocation of P-ERK1/2. PMID- 19218115 TI - [Chorionic villus cell culture and karyotype analysis in 110 cases of early spontaneous abortion]. AB - OBJECTIVE: To investigate the value of fetal chromosomal karyotype analysis in cases of early spontaneous abortion. METHODS: Chorionic villus specimens obtained from 110 cases of early spontaneous abortion were cultured for karyotype analysis. RESULTS: Of the 110 cases, chorionic villus was successfully cultured in 103 cases (93.7%), and abnormal embryo karyotypes were identified in 52 cases (50.5%). Trisomy was the most frequent embryo karyotype abnormalities in these cases, and chromosomal aberration occurred in 29 cases (52.9%) of the first spontaneous abortion and in 23 cases (42.6%) of repeated abortions. Female fetuses accounted for 75.5% (78 cases) in the spontaneously aborted fetuses and for 67.3% (35 cases) in fetuses with chromosomal abnormalities. CONCLUSION: Embryo chromosomal abnormality is the most important reason of early spontaneous abortion, and karyotype analysis of the villus helps identify the causes of abortion and ensure the success of the next pregnancy. PMID- 19218116 TI - [Expression of vascular endothelial growth factor and its receptor Flk-1/KDR in benign and malignant breast neoplasms]. AB - OBJECTIVE: To investigate the differences in the expression of angiogenesis related molecules between benign and malignant breast neoplasms. METHODS: Thirty breast cancer patients (33 foci) and 30 with benign breast neoplasms (34 foci) were examined for CD34, vascular endothelial growth factor (VEGF) and Flk-1/KDR expressions using immunohistochemistry. RESULTS: In patients with breast cancer, the microvessels densely distributed around the cancer nest. The microvessel density (MVD) in the cancer patients was significantly higher than that in patients with benign tumors (34.48+/-8.34 vs 18.65+/-4.69, P<0.05). In the breast cancer patients, strong VEGF expression was found in the epithelial cells and vascular endothelial cells around the breast carcinoma, and Flk-1/KDR was also strongly expressed in the vascular endothelial cells. The expressions of VEGF and Flk-1/KDR were hardly detectable in the benign tumors. CONCLUSION: VEGF is an important regulatory factor in promoting breast tumor angiogenesis. PMID- 19218117 TI - [Effect of recombinant caspase-3 gene transfection on CNE2 cell apoptosis]. AB - OBJECTIVE: To investigate the apoptosis-inducing effect of caspase-3 on human nasopharyngeal carcinoma cells (CNE2). METHODS: Recombinant caspase-3 was subcloned into the eukaryotic expression vector PEGFP-C1 containing the reporter gene using DNA recombinant technique. CNE2 cells were transfected with the recombinant caspase-3 gene via lipofectamine 2000 and the expression of caspase-3 mRNA was detected by reverse transcription-polymerase chain reaction. The cell morphological changes were observed under fluorescence microscope and electron microscope and the cell survival rate after the transfection was assessed by MTT assay. RESULTS: Transfection with the recombinant caspase-3 gene induced significant apoptosis in CNE2 cells, which exhibited obvious morphological changes typical of apoptotic cells. CONCLUSION: The recombinant caspase-3 gene can inhibit the growth and effectively induce apoptosis of CNE2 cells. PMID- 19218118 TI - [Interface shear stress between the artificial bones and injectable calcium phosphate glue: an experimental study in rabbits]. AB - OBJECTIVE: To evaluate the effect of the composite bone material, fibrin glue (FG) combined with beta-tricalcium phosphate (beta-TCP)/monocalcium phosphate, in repairing bone defects and assess the feasibility of using this cement for artificial joint fixation. METHODS: Bone defects were induced in 16 normal adult New Zealand white rabbits at the bilateral femoral lateral condyles where an 8-mm deep hole (4 mm in diameter) was drilled on each side. The composite FG/calcium phosphate cement (CPC) (solution: power volume ratio of 0.3:1) was injected on one side of the bone defects (experimental group) and pressurized for 10 minutes, and CPC was injected on the other side (control). The rabbits were sacrificed at 2, 4, 8 or 12 weeks after the operation for gross observation and biomechanical tests. RESULT: The composite material FG/CPC was more effective than CPC for bone defect repair, and biomechanical tests revealed significant differences between them (P<0.05). The shear stress of the artificial bone cement in the FG/CPC group was stronger than that in CPC group. CONCLUSION: The FG/CPC composite possesses good biocompatibility and osteoconductivity and may serve as an ideal material for repairing bone defects. PMID- 19218119 TI - [Distribution and drug resistance analysis of bacteria in different wound infections]. AB - OBJECTIVE: To analyze the distribution and drug resistance of bacteria in different wound infections and provide evidence for wound infection control in subtropical regions. METHODS: This study involved 265 patients from 4 different departments of our hospital who experienced wound infections between July, 2007 and July, 2008. The bacterial strain distribution in the wounds and drug resistance of the bacteria were analyzed. RESULTS: Acinetobacter baumanii (39% of the total strain identified) was the most frequent bacterial strain causing infection of the burn wounds, followed by Proteus mirabilis (20%) and Pseudomonas aeruginosa (20%). E. coli infection was prevalent in the departments of general surgery (37%) and urinary surgery (64%), and Pseudomonas aeruginosa and Pseudomonas pneumonia infections were detected at the rate of 30% and 43% in the urinary surgery department, respectively. Different bacterial strains were found at similar rates around 10% in the wounds of patients undergoing traumatic surgery. CONCLUSION: Despite that the commonly seen pathogenic bacteria in burn patients including Staphylococcus aureus have been effectively controlled by early application of antibiotics, the opportunistic pathogens such as Acinetobacter baumanii and Proteus mirabilis often survive these antibiotics, and some strains evolve to be drug-resistant and even multi-drug-resistant. E. coli infection is prevalent in general surgery and urinary surgery departments, where Staphylococcus aureus and Pseudomonas aeruginosa infections can also be found frequently. All kinds of bacteria infection are present in trauma surgery department, each found at the rate around 10%. PMID- 19218120 TI - [Development of time-resolved fluoroimmunoassay kit for detection of IgM antibodies against hepatitis B core antigen]. AB - OBJECTIVE: To prepare a time-resolved fluoroimmunoassay (TRFIA) kit for clinical detection of IgM antibodies to hepatitis B core antigen (HBc). METHODS: Immunocapture method was used to develop the TRFIA kit for detection of the anti HBc IgM antibodies, and the precision, cross-reactivity and sensitivity of the kit were tested with the clinical serum samples. RESULTS: The intra- and inter assay coefficients of variation of the TRFIA kit were 4.8%-7.2% and 7.5%-8.6%, respectively, and no cross-reactivity with anti-HBs, anti-HBc-IgG or anti-HBe was found. Comparison of the results of the TRFIA kit and enzyme-linked immunosorbent assay (ELISA) demonstrated greater sensitivity of the kit than ELISA in detecting the anti-HBc IgM antibodies in 584 serum samples. According to the detection results in 300 serum samples from healthy donors, the cutoff value of the TRFIA kit was 4.5 times of the fluorescence value of the negative control. CONCLUSION: This TRFIA kit for detecting anti-HBc IgM antibodies meets the demand for clinical application and can replace the ELISA kits. PMID- 19218121 TI - [Expression of TBX3 mRNA and its role in the pathogenesis and metastasis of breast cancer]. AB - OBJECTIVE: To explore the role of TBX3 gene in the pathogenesis of breast cancer. METHODS: The total RNA of 51 fresh breast cancer tissues and the corresponding adjacent tissues were extracted and reverse transcribed into cDNA to detect the expression of TBX3 mRNA by real-time PCR. The correlation between TBX3 mRNA expression and the clinicopathologic parameters in relation to breast cancer metastasis was analyzed. RESULT: Compared to that in the adjacent tissues, the expression of TBX3 mRNA was markedly increased in breast cancer tissues. TBX3 mRNA expression was significantly higher in metastatic breast cancer than in non metastatic tumors. CONCLUSION: Increased expression of TBX3 mRNA suggests the involvement of TBX3 in the pathogenesis and metastasis of breast cancer. PMID- 19218122 TI - [Effect of intense pulsed light on transforming growth factor-beta1 mRNA expression in rat skin]. AB - OBJECTIVE: To observe the effect of intense pulsed light (IPL) on transforming growth factor-beta1 mRNA (TGF-beta1 mRNA) expression in rat skin and explore the molecular mechanisms of photorejuvenation. METHODS: Fifteen SD rats were exposed to IPL in 3 dermal regions with triple pulses (duration of 4, 5, and 6 ms) at the energy density of 34 J/cm2 and pulse delay of 20 or 25 ms. On days 1, 3, 5, 7, 15, and 30 after the treatment, skin specimens from the treated and non-treated areas were obtained to detect TGF-beta1 mRNA expression with in situ hybridization. RESULTS: In the UPL-exposed skin areas, TGF-beta1 mRNA expression was detected in the epidermal keratinocytes and dermal cells 1 day after the exposure, reaching the highest expression level on day 7 followed by gradual decrement since day 15, and till day 30, only weak expression was found in the dermal cells. In the non-exposed regions, the cells remained negative for TGF beta1 mRNA. CONCLUSION: IPL can enhance TGF-beta1 mRNA expression in the skin, suggesting that TGF-beta1 plays an important role in dermal remodeling in photorejuvenation. PMID- 19218123 TI - [Effect of propofol at different effect-site concentrations on approximate entropy of transient evoked otoacoustic emission signals in adults]. AB - OBJECTIVE: To study the effect of propofol at different effect-site concentrations on approximate entropy (ApEn) of transient evoked otoacoustic emission (TEOAE) signals in adults and investigate the possibility of using ApEn for monitoring anesthesia depth. METHODS: Fifteen ASA class I or II patients (aged 18-49 years with normal hearing) undergoing elective surgery under general anesthesia were enrolled in this study. Anesthesia was maintained with target controlled infusion of propofol. With the effect-site concentrations of 1, 2, 3 and 4 microg/ml, TEOAE signals were monitored and recorded before and after anesthesia. ApEn of TEOAE in 4 frequency ranges (0-2, 1-3, 2.5-4.5, and 4-6 kHz) were calculated using MATLAB software. RESULTS: The ApEn of TEOAE in different frequency ranges showed no significant differences at the same effect-site concentration of propofol, or at different effect-site concentrations in the same frequency range (P>0.05). CONCLUSION: Anesthesia with propofol at different effect-site concentrations does not obviously affect ApEn of TEOAE signals in adults, and ApEn can not be used as the indicator for evaluating the depth of anesthesia. PMID- 19218124 TI - [MTT assay for detecting 5-fluorouracil chemosensitivity of human breast carcinoma cell line]. AB - OBJECTIVE: To assess the feasibility of MTT colorimetric assay for testing the in vitro chemosensitivity of breast cancer cells to 5 fluorouracil (5-Fu). METHODS: The chemosensitivity of human breast carcinoma cell lines MCF-7 and MDA-MB-435S to 5-Fu at different concentrations was evaluated with MTT assay. RESULTS: 5-FU treatment resulted in dose-dependent growth inhibition of the breast cancer cells with both low and high metastatic capacities. CONCLUSIONS: MTT assay may help select appropriate chemotherapeutic agents and provides evidence for individualized chemotherapy for breast cancer. PMID- 19218125 TI - [Clinical application of prefilled pen and conventional syringe during controlled ovarian stimulation for in vitro fertilization]. AB - OBJECTIVE: To investigate the efficacy, convenience and costs of recombinant follitropin alpha administered by a prefilled pen device and conventional syringe in Chinese women undergoing controlled ovarian stimulation for in vitro fertilization (IVF). METHODS: A total of 184 patients undergoing IVF treatment were enrolled in this study. According to a long-term recombinant follicle stimulating hormone (rFSH) protocol, ovarian stimulation was performed with the prefilled pen and conventional syringe at random in these subjects, and the dose of follitropin, number of oocytes and embryo parameters and IVF-ET outcome were compared between the two groups. RESULTS: The total rFSH dose, cost, and frequency of hospital visits were significantly lower in the pen protocol group, but the residual rFSH amount was higher. Compared with conventional injections, the prefilled pen was associated with significantly lowered rate of local redness, high rate of local bruise, more frequent follitropin dose modulation and lower serum oestradiol levels on HCG day. No significant difference was found in the endometrial thickness, numbers of oocytes retrieved, MII oocytes, transferred embryo, or the clinical pregnancy rates between the two groups. The ratio of MII oocytes, good quality embryo rates and implantation rates was significantly higher in the pen group with lower incidences of moderate and severe ovarian hyperstimulation syndrome. CONCLUSION: The prefilled pen provides an easy, safe, effective and more patient-friendly means for controlled ovarian stimulation procedure in Chinese women, but more attention should be given to protocol optimization and patient education. PMID- 19218126 TI - [Laparoscopic radical cystectomy and detenial sigmoid colon orthotopic neobladder reconstruction for bladder tumor in a 3-year-old boy]. AB - OBJECTIVE: To present a case of laparoscopic radical cystectomy and detenial sigmoid colon orthotopic neobladder reconstruction for bladder tumor in a child. METHODS: A 3-year-old boy with bladder rhabdomyosarcoma underwent laparoscopic radical cystectomy and detenial sigmoid colon orthotopic neobladder reconstruction. The bilateral pelvic lymphadenectomy and cystectomy were performed laparoscopically, and removal of the mobilized specimens and urinary diversion operation were managed through enlarged abdomen incision. The urinary diversion procedure included construction of the detenial sigmoid pouch, bilateral stented antiflux implantation of the ureters in the pouch and orthotopic anastomosis of the neobladder with the urethra. RESULTS: The total operative time was 6 h, and the laparoscopic procedure lasted for about 3.5 h. The intraoperative blood loss was 50 ml, and 200 ml concentrated red blood cell transfusion was used for the safety of the patient. Six dissected lymph nodes in each pelvic side and the surgical margins of the ureter and urethra were all free of tumor invasion. Bowel peristalsis recovered 3 days after the operation, and the pelvic drainage and the neobladder drainage tubes were removed on day 7 and 14, respectively. The urethral catheter and ureteral stents were removed 25 days after the operation. The daytime urine control and micturition recovered 1 week after the operation. The neobladder capacity was about 110 ml, with residual urine volume of 10 ml and peak flow rate of 12 ml/s after 5 months. No perioperative complications occurred such as water-electrolyte and metabolic acid base balance disorders, urinary leakage, reflux or bowel obstruction. CONCLUSION: Laparoscopic radical cystectomy is minimally invasive, reduces intraoperative blood loss and allows rapid postoperative recovery, and can be a promising approach to management of bladder rhabdomyosarcoma in children. PMID- 19218127 TI - [Analysis of natural killer cell immunoglobulin-like receptor genes in Chinese]. AB - OBJECTIVE: To investigate the genotypes of natural killer cell immunoglobulin like receptor (KIR) genes and their frequencies in Chinese subjects and explore the mechanism of the actions of nature killer cells. METHODS: The DNA samples were obtained from 67 randomly selected unrelated Chinese Han individuals for genotyping of the KIR genes using PCR with sequence-specific primers (PCR-SSP), and the frequencies of the KIR genes in these Chinese subjects were compared with the reported frequencies in populations of other nationalities. RESULTS: Sixteen KIR genes were identified in these Chinese subjects, and 87.5% of these genes were expressed at frequencies above 0.35. Fourteen functional KIR genes combined into 25 KIR genotypes, among which the most frequent genotype KIR-2DL1-2DL3-2DL4 3DL1-3DL2-3DL3-2DS4 showed a frequency of 0.373, while the frequencies of all the other genotypes were no greater than 0.09. Comparison of the KIR combinations in Chinese Han population with those of Japanese, Korean, and Caucasians populations identified 8.93% of the KIR combinations shared by all these populations; the Chinese, Koreans and Caucasians shared 5.36% common KIR combinations, whereas only 1.79% common combinations were found in Chinese and Caucasians. In this study, 16 new gene combinations were identified (25.28%). CONCLUSION: This study shows the high-frequency distribution of a single KIR gene polymorphism. The KIR combination KIR-2DL1-2DL3-2DL4-3DL1-3DL2-3DL3-2DS4 has the highest frequency in Chinese, Japanese, Korean and Caucasian populations, indicating that inhibitory signal transduction pathway plays an important role in the function of the natural killer cells. This study provide clues for new approaches for improving the prognosis of kidney transplantation by enhancing or inhibiting the function of the natural killer cells instead of life-time usage of immunosuppressive agents. PMID- 19218128 TI - [Modified neurological tests for evaluating the severity of cerebral infarction and estimating the infarct area in rats]. AB - OBJECTIVE: To test the reliability of quantitative neurological tests in evaluating the neurological deficits in rat models of cerebral ischemia. METHODS: Forty male SD rats (7 to 9 weeks old) were subjected to transient (1 h) middle cerebral artery occlusion (MCAO) to induce cerebral infarction and 9 received sham operation only. The motor and sensory functions of the rats were measured before and 1, 3, 7 and 14 days after MCAO by accelerating rotarod test and modified adhesive removal test. Fourteen days later, the rats were sacrificed for histological examination, and the lesion size was determined by Winroof image analysis. The neurological deficits of the rats with different lesion sizes and at different time points were analyzed. RESULTS: According to the infarct size, the rats after MCAO were divided into two subgroups with large lesions [(68.56+/ 12.54)%] affecting both the cortical area and basal ganglia and small lesions [(12.45+/-9.06)%] affecting the basal ganglia. Significant differences in the results of modified adhesive removal test and rotarod test were found between the two groups, and a liner relationship was noted between the lesion size and the results of the neurological tests. CONCLUSION: The modified quantitative neurological tests can reliably evaluate the severity of the infarction and estimate the infarct size in rats with cerebral infarction. PMID- 19218129 TI - [Pilot-scale production and quality control of multiepitope hepatitis B virus DNA vaccine]. AB - OBJECTIVE: To establish a method for pilot-scale production and quality control of multiepitope hepatitis B virus (HBV) DNA vaccine (PVAX-HS). METHODS: Recombinant DH5alpha/pVAX-HS was obtained by fed-batch fermentation, and the plasmid was extracted by alkaline lysis and concentrated by ultrafiltration. The plasmid DNA was purified by a three-step column chromatography to obtain the DNA vaccine, and quality control tests were performed on the final product. RESULTS: The quantity of the fed-batch product reached 50-60 g/L, and the final plasmid output was 1.0 mg per gram of the bacteria. The quality of the DNA vaccine met the requirements for medical use. CONCLUSION: A simple and stable procedure was established for pilot-scale production of multiepitope HBV DNA vaccine, which allows potential large-scale production of the DNA vaccine. PMID- 19218130 TI - [Application of ultrathin transnasal gastroscopy in percutaneous endoscopic gastrostomy]. AB - OBJECTIVE: To assess the application of ultrathin transnasal gastroscopy in transnasal percutaneous endoscopic gastrostomy (nPEG) in patients with trimus and/or pharyngeal narrowing. METHODS: Nine consecutive patients underwent PEG with the Introducer method using conventional gastroscopy (5 cases) or ultrathin transnasal gastroscopy (4 cases). Among the 4 patients undergoing nPEG, 3 received radiotherapy for nasopharyngeal cancer, including two with trimus and one with trimus and pharyngeal narrowing. The procedure time, safety, discomfort and complications in these cases were compared and evaluated. RESULTS: The average procedure time of PEG was 17+/-3.5 min in conventional gastroscopy group and 17+/-3.1 min in nPEG group. No complications were observed in these patients, but the patients in nPEG group reported less discomfort associated with the procedure. CONCLUSIONS: Ultrathin transnasal gastroscopy reduces the discomfort of the procedure and is safer than conventional gastroscopy for PEG, especially in patients with trimus and/or pharyngeal narrowing. PMID- 19218131 TI - [Urethral acellular matrix graft for repairing urethral defect in rabbits]. AB - OBJECTIVE: To assess the biocompatibility of a urethral acellular matrix graft (UAMG) and evaluate its effect in repairing urethral defect in rabbit models. METHODS: The UAMG was prepared and its structural features were observed using optical and electron microscopy. In vitro cultured rabbit bladder smooth muscle cells were seeded on UAMG and the cell proliferation was observed. The cytotoxicity of the aqueous extract of the UAMG against the cells was evaluated by MTT assay, and its biocompatibility was assessed by implanting the grafts subcutaneously on the back of the rabbits. In 24 male rabbits, a 2-cm urethral defect was induced and repaired with UAMG (experimental group, n=12) or left untreated (control group, n=12). In both groups, the rabbits were sacrificed 2, 4, 8 and 12 weeks after the operation for histological and immunohistochemical examination of the tissue regeneration. RESULTS: The UAMG had a reticular fibrous structure without cell residues. The bladder smooth muscle cells showed normal proliferation on UAMG with normal cell morphology. The rabbits receiving the implants showed no abnormal response, and the UAMGs gradually degraded in vivo with grade 0 or 1 cytotoxcity showing satisfactory cytocompatibility. In the experimental group, new urethral tissues that were histologically compatible with normal urethral tissues were regenerated in the defect area 12 weeks after UAMG implantation. CONCLUSION: As a tissue engineered scaffold material for urethral reconstruction, the UAMG possesses good biocompatibility and can induce the regeneration of urethral epithelial cells and smooth muscle cells. PMID- 19218132 TI - [Efficacy and safety of segmental pulmonary vein isolation and circumferential pulmonary vein isolation in patients with atrial fibrillation: a comparative study]. AB - OBJECTIVE: To compare the efficacy and safety of segmental pulmonary vein isolation (SPVI) and circumferential pulmonary vein isolation (CPVI) guided by EnSite NavX system in patients with atrial fibrillation (AF). METHODS: Eighty five patients with paroxysmal AF and persistent AF were enrolled in this study. Forty patients (30 with paroxysmal AF and 10 with persistent AF) underwent SPVI procedure, and 45 (31 with paroxysmal AF and 14 with persistent AF) underwent CPVA guided by EnSite NavX three-dimensional electrophysiological mapping system. All the patients were followed up for over six months. RESULTS: The success rate was 65% in the SPVI group and 84.4% in the CPVI group (P=0.0332), with incidence of major complications of 17.5% and 6.7%, respectively (P=0.0845). In the SPVI group, 12.5% patients had pulmonary vein stenosis after the operation, which occurred in none of the patients in the CPVI group (P=0.0312). The total procedure time was 200.4+/-37.0 min in the SPVI group, significantly shorter than that in the CPVI group (226.5+/-26.1 min, P=0.002). The fluoroscopy time in the SPVI group was obviously longer than that in the CPVI group (54.7+/-9.7 vs 27.1+/ 3.1 min, P<0.001). CONCLUSIONS: CPVI guided by EnSite NavX system is more effective than SPVI for treatment of atrial fibrillation with significantly shortened fluoroscopy time but prolonged procedure time. The two procedures results in comparable incidences of major complications, but CPVI is associated with reduced rate of pulmonary vein stenosis in comparison with SPVI. PMID- 19218133 TI - [Dynamic enhanced scanning of the pulmonary nodules with 16-slice spiral CT]. AB - OBJECTIVE: To assess the value of dynamic enhanced CT scanning in diagnosis of pulmonary nodules. METHODS: Sixty-nine patients with pulmonary nodules underwent examination with dynamic enhanced CT scanning. Of these patients, 53 with definite diagnoses confirmed by histological or clinical data were divided into three groups according to the nature of the nodules, namely the malignant group (n=34), benign group (n=13) and active inflammatory group (n=6). The time-density curve (T-DC) and the corresponding parameters of these patients were compared. RESULTS: Significant differences in the net enhancement and S/A ratio were found among the 3 groups (P<0.001). The active inflammatory nodules showed the highest enhancement, followed by malignant nodules and then by benign nodules, with significant differences between any of the two groups (P<0.05). At the diagnostic threshold of net enhancement by 25 Hu or S/A ratio by 10% for malignant nodules, a higher negative predictive value and accuracy were obtained. The difference in the wash-out value among the 3 groups was statistically significant (P<0.05). When diagnostic criteria for malignancy was defined by a wash-in value of 25 Hu or higher and wash-out value of 0-35 Hu, the false-positive rate was decreased from 57.89% to 47.37%. CONCLUSION: The application of 16-slice spiral CT and its CT perfusion software allows convenient dynamic enhancement study of pulmonary nodules and can be helpful in their differential diagnosis. PMID- 19218134 TI - [Role of CD4+CD25+Foxp3+ regulatory T cells in type 2 diabetic nephropathy]. AB - OBJECTIVE: To investigate the role of CD4+CD25+Foxp3+ regulatory T cells (Treg) in type 2 diabetic nephropathy and provide new clues for prevention and treatment of type 2 diabetic nephropathy. METHODS: Flow cytometry was used to analyze the expression rate of CD4+CD25+Foxp3+ Treg cells in 60 patients with type 2 diabetes and 15 normal subjects. RESULTS: No significant difference was found in the expression rate of CD4+CD25+Foxp3+ Treg cells between the control group and the type 2 diabetic patients. In the type 2 diabetic patients with microalbuminuria and macroalbuminuria, the expression of CD4+CD25+Foxp3+ Treg cells was significantly lowered in comparison with that in the control group (P<0.05), and patients with macroalbuminuria showed significantly lower expression of CD4+CD25+Foxp3+ Treg cells than the microalbuminuric patinets (P<0.05). Significant inverse correlations were noted between the disease course and the expression of CD4+CD25+Foxp3+ Treg cells and between the urinary albumin excretion rate (UAER) and the expression of CD4+CD25+Foxp3+ Treg cells. CONCLUSION: CD4+CD25+Foxp3+ Treg cells may play a role in the occurrence and development of type 2 diabetic nephropathy. PMID- 19218135 TI - [Graph-based interactive three-dimensional segmentation of magnetic resonance images of brain tumors]. AB - We propose a graph-based three-dimensional (3D) algorithm to automatically segment brain tumors from magnetic resonance images (MRI). The algorithm uses minimum s/t cut criteria to obtain a global optimal result of objective function formed according to Markov Random Field Model and Maximum a posteriori (MAP-MRF) theory, and by combining the expectation-maximization (EM) algorithm to estimate the parameters of mixed Gaussian model for normal brain and tumor tissues. 3D segmentation results of brain tumors are fast achieved by our algorithm. The validation of the algorithm was tested and showed good accuracy and adaptation under simple interactions with the physicians. PMID- 19218136 TI - [In vitro tracing of rat bone marrow mesenchymal stem cells using the fluorescent dye CFSE]. AB - OBJECTIVE: To determine the optimal condition for labeling rat mesenchymal stem cells (MSCs) using the fluorescent dye CFSE and the maximum time length allowed by CFSE staining for MSC tracing in vitro. METHODS: Rat MSCs were labeled with CFSE at different concentrations (2.5, 5.0, 10.0, 20.0 and 40.0 micromol/L) for 1, 5 or 10 min. The transfection efficiency and fluorescence intensity in the cells were measured by flow cytometry and fluorescence microscope to determine the optimal condition for MSC labeling. Under the optimal condition, the effect of CFSE on the growth of MSCs was evaluated by MTT assay, and flow cytometry and fluorescence microscope were used to determine the maximum time length following CFSE labeling to allow MSC tracing. RESULT AND CONCLUSION: Staining with CFSE at 20.0 micromol/L for 5 min was optimal for labeling rat MSCs in vitro, which allowed detection of the MSCs as long as 21 days after the labeling without obviously affecting the cell growth (P>0.05). PMID- 19218137 TI - [Expression of Beclin1 in primary hepatocellular carcinoma]. AB - OBJECTIVE: To detect Beclin1 expression and explore its clinical significance in primary hepatocellular carcinoma (HCC). METHODS: Beclin1 expressions in 10 normal hepatic tissues, 30 hepatitis liver, 30 cirrhotic liver and 50 HCC tissues were detected by immunohistochemical staining. RESULTS: The positivity rates of Beclin1 expression in the HCC, cirrhotic liver, hepatitis liver and normal liver tissues were 78.00% (39/50), 26.67% (8/30), 53.33% (16/30), and 10.00% (1/10), respectively, showing significant differences between them (chi(2)=28.31, P<0.05). Beclin1 expression was significantly higher in HCC tissues than in the cirrhotic, hepatitis and normal liver tissues (chi(2)=20.39, 5.31, and 14.41, respectively, P<0.05), and hepatitis tissues showed significantly higher Beclin1 expression than hepatic cirrhosis tissues and normal hepatic tissues (chi(2)=4.44 and 4.12, respectively, P<0.05). CONCLUSION: The abnormal expression of Beclin1 is closely associated with the pathogenesis and development of primary hepatocellular carcinoma, and may play an important role in this process. PMID- 19218138 TI - [Effects of phosphocreatine on plasma brain natriuretic peptide level in elderly patients with chronic congestive heart failure]. AB - OBJECTIVE: To investigate the therapeutic effects of phosphocreatine in elderly patients with chronic congestive heart failure (CHF) and its effects on plasma brain natriuretic peptide (BNP). METHODS: Forty elderly patients with chronic CHF were randomly divided into two groups to receive basic treatment (control group) and additional phosphocreatine treatment (treatment group) with a treatment course of 8 weeks. The patients were evaluated for improvement in New York Heart Association (NYHA) functional class, symptoms, left ventricular end-diastolic diameter (LVEDD), left ventricular end-systolic diameter (LVESD), and left ventricular ejection fraction (LVEF) and the levels of BNP before and after treatment. RESULTS: After 8 weeks of treatment, the overall efficacy rate was significantly higher in treatment group than in the control group, and LVESD, LVEDD, LVEF and BNP level of the treatment group were significantly lowered in comparison with those of the control group (P<0.05). CONCLUSION: Phosphocreatine in addition to the basic treatment can reduce the BNP level and improve the cardiac systolic and diastolic function in elderly patients with chronic CHF. PMID- 19218139 TI - [Survivin and COX-2 expressions in giant cell tumor of bone and their relation to the prognosis]. AB - OBJECTIVE: To investigate the expression of survivin and COX-2 in giant cell tumor of bone (GCT) and explore the prognostic factors for GCT. METHODS: The expressions of survivin and COX-2 in 39 GCT tissues of three Jaffe grades and 4 normal bone tissues were detected by immunohistochemical staining, and the data were analyzed in relation to the clinicopathological features of the patients. RESULTS: The expressions of survivin and COX-2 were significantly higher in the GCT tissues than in normal bone tissues (P<0.01). A positive correlation was found between survivin and COX-2 expressions and the pathological grade (P<0.01), but their expressions were not correlated to the patients' gender, age or surgical approaches (P>0.05). An obviously lowered recurrence rate was observed in patients with resection of the bone segment compromised by the tumor and subsequent bone grafting. Survivin and COX-2 were not independent risk factors of the prognosis of GCT. CONCLUSION: Survivin and COX-2 expressions may participate in the pathogenesis and development of GCT, but is not indicative of the prognosis. PMID- 19218140 TI - [Relationship between protein oxidation levels in the follicular fluid and the outcome parameters of in vitro fertilization-embryo transplantation]. AB - OBJECTIVE: To explore the relationship between protein oxidation levels in the follicular fluid and the outcome parameters of in vitro fertilization-embryo transplantation (IVF-ET). METHODS: The levels of advanced oxidation protein products (AOPP) in the follicular fluid were measured in 64 women with tubal infertility undergoing IVF-ET. The relationship between the AOPP levels and IVF ET outcome parameters was analyzed. RESULTS: AOPP levels showed significant inverse correlations between the proportion of mature oocytes (r=-0.401, P=0.001), fertilization rate (r=-0.257, P=0.045), cleavage rate (r=-0.290, P=0.024) and good embryo rate (r=-0.520, P=0.000). AOPP levels differed significantly between the groups with different retrieved oocyte numbers (F=3.851, P=0.027), being the lowest in women with 8 to 15 retrieved oocytes and the highest in those with retrieved oocytes below 8. The AOPP level in the non pregnant women was significantly higher than that in the pregnant women (t=3.665, P=0.001). The AOPP levels also differed significantly with age (F=15.919, P=0.000), and the women >35 years of age had the highest level and those below 30 years had the lowest level. CONCLUSION: Protein oxidative stress is present in the follicular fluid of women on IVF-ET cycles. High level of AOPP may have adverse effects on the oocytes and early embryonic development and may affect the outcome of IVF-ET. PMID- 19218141 TI - [Expression of Notch1 on peripheral lymphocytes before and after acute graft rejection following renal transplantation]. AB - OBJECTIVE: To study the changes in Notch1 expression on peripheral lymphocytes after acute graft rejection after renal transplantation. METHODS: Twenty renal transplant recipients experiencing acute graft rejection and 20 without acute rejection were enrolled in this study. Flow cytometry was used to detect the expression of Notch1 on peripheral lymphocytes of the patients before operation, at the occurrence of acute rejection and after anti-rejection therapy. The rates of Notch1-positive lymphocytes measured at different time points were compared between the two groups. RESULT: In patients with acute graft rejection, Notch1 expression at the time of rejection onset was significantly higher than that before operation (t=4.245, P=0.000) and that of patients with graft rejection (t=3.839, P=0.000), and was obviously decreased after anti-rejection therapy (t=3.102, P=0.004). Patients without graft rejection showed no significant changes in Notch1 expression after the transplantation (P=0.409). Notch1 expression was comparable between the recipients receiving Tac therapy and those with CsA therapy (P>0.05). CONCLUSION: Monitoring Notch1 expression on the peripheral lymphocytes after renal transplantation may help in the diagnosis of acute graft rejection and prediction of the effect of an anti-rejection therapy. PMID- 19218142 TI - [Effects of partial revascularization in elderly patients over 80 years old with coronary artery diseases]. AB - OBJECTIVE: To observe the clinical effects of partial revascularization on elderly patients with coronary artery diseases(CAD) METHODS: Percutaneous coronary interventions (PCI) of the most likely culprit vessels were performed in 37 patients over 80 years old with multivessel coronary artery diseases (CAD). RESULTS: The success rate of PCI was 100% in these elderly patients without serious complications. The in-hospital mortality rate was 2.7%, and all the other patients recovered and were discharged. CONCLUSION: Partial revascularization for elderly CAD patients can achieve satisfactory clinical results, and close attention should be given to the heart and kidney function of these patients. PMID- 19218144 TI - [Preoperative revascularization in high-risk patients undergoing vascular surgery]. AB - Patients undergoing vascular surgery are at increased risk for cardiac complications related to the presence of underlying coronary artery disease. Preoperative cardiac evaluation may help to identify high-risk patients in whom coronary angiography may be planned with subsequent coronary revascularization for the purpose of improving perioperative and long-term cardiac outcomes. However, the indications and efficacy for type of revascularization for the reduction of cardiac complications compared to medical therapy have been controversial. The aim of the review was to summarize the role of preoperative revascularization compared to conservative medical therapy before elective vascular surgery using current evidence from published studies. PMID- 19218145 TI - [Diarrhea from the infectologist's point of view]. AB - Gastroenteritis is a nonspecific term for various pathologic states of the gastrointestinal tract. Gastroenteritis causing pathogens are the second leading cause of morbidity and mortality worldwide. In the developed countries diarrhea is the most common reason for missing work, while in the developing world, it is a leading cause of death. Internationally, the mortality rate is 5-10 million deaths each year. "Traveller's diarrhea" is a polyetiologic common health problem of international travellers which affects travellers generally for days, but it can result in chronic postinfectious irritable bowel syndrome as well. Infectious agents usually cause acute gastroenteritis either by adherence of the intestinal mucosa, or by mucosal invasion, enterotoxin production, and/or cytotoxin production. The incubation period can often suggest the cause of etiology. When symptoms occur within 6 hours of eating, ingestion of preformed toxin of S. aureus or Bacillus cereus should be suspected. The incidence of hypervirulent C. difficile associated colitis is an emerging problem as a healthcare system associated infection. While infectious agents do not commonly cause chronic diarrhea, those that do include C. difficile, Giardia lamblia, Entamoeba histolytica, Cryptosporidium, Aeromonas and Yersinia . Amoebiasis is the second to malaria as a protozoal cause of death. Infection with HIV is also a common cause of diarrhea. PMID- 19218146 TI - [Off-label therapies in oncology]. AB - The unapproved (off-label) therapies represent a special problem in oncology since they are in the borderline of legal regulation and of free medical practice. Although in Hungary the off-label therapies were regarded as clinical trials without permission until the new regulation came into action at the end of October 2008, certain experts were even so arguing for its use because clinical practice changes more rapidly than the prescription label. Moreover, manufacturers are not obliged to submit Supplemental New Drug Application in spite of the fact that enough evidence is generated to do so. The regulation of off-label therapies should meet the conditions of free medical practice, evidence based medicine, demand of patients for new chances and expectations of regulatory authorities. In this paper, following the criticism of the Hungarian status and the new regulation, as well as the review of international practice, the author outlines the frame of a potential regulation separately indicating the role of practicing physicians and authorities. PMID- 19218147 TI - [Akt enzyme: new therapeutic target in cancer and diabetes?]. AB - Alteration of apoptotic processes plays a central role in the development and progression of several chronic disorders. Proteins responsible for the regulation of apoptosis are therapeutic targets; these include the Akt enzyme. Akt enzyme is expressed in most cell types. Akt activation is regulated by growth factors, insulin, and also environmental factors as altered oxygen tension and high temperature. Akt is a central regulator of cellular metabolism and survival. Akt function is reportedly altered in some disorders. An increased activity of Akt has been described in prostate, breast, colon, and pancreatic cancer, as well as in hematological malignancies. Akt is also a factor in the pathomechanism of diabetes as it determines beta-cell apoptosis of Langerhans islets and insulin sensitivity of the cells. Several studies revealed that some of the marketed drugs including statins, thiazolidinediones and ACE inhibitors modulate Akt activity. There are efforts to develop specific Akt inhibitors that may improve the efficacy of chemotherapy. Triciribine and perifosine are two Akt inhibitors in developmental phase 1 and 2 that may improve survival in breast cancer, pancreas cancer, gastrointestinal stroma tumor, sarcoma and melanoma, and in hematological malignancy. PMID- 19218148 TI - [The therapeutical practice of the mathematician Farkas Bolyai (1775-1856)]. PMID- 19218150 TI - Delay effects in brain dynamics. Introduction. AB - This brief introductory paper reviews the methods and the results presented in the special issue. The general destabilizing effects of time delays in nonlinear dynamical systems are summarized and some similarities in the philosophical approaches of neural systems research in distinct disciplines are pointed out. All the invited papers focus on the central role of time delays in the dynamics of neural systems. The research contributions are set in order according to the increasing number of neurons involved in the corresponding study from a couple of neurons through neural fields to populations and clusters of neurons. PMID- 19218151 TI - Compensating time delays with neural predictions: are predictions sensory or motor? AB - Neural delays are a general property of computations carried out by neural circuits. Delays are a natural consequence of temporal summation and coding used by the nervous system to integrate information from multiple resources. For adaptive behaviour, however, these delays must be compensated. In order to sense and interact with moving objects, for example, the visual system must predict the future position of the object to compensate for delays. In this paper, we address two critical questions concerning the implementation of the compensation mechanisms in the brain, namely, where does compensation occur and how is it realized. We present evidence showing that compensation can happen in both the motor and sensory systems, and that compensation using 'diagonal neural pathways' is a suitable strategy for implementing compensation in the visual system. In this strategy, neural signals in the early stage of information processing are sent to the future cortical positions that correspond to the distance the object will travel in the period of transmission delay. We propose a computational model to elucidate this using the retinal visual information pathway. PMID- 19218152 TI - Time-delayed feedback in neurosystems. AB - The influence of time delay in systems of two coupled excitable neurons is studied in the framework of the FitzHugh-Nagumo model. A time delay can occur in the coupling between neurons or in a self-feedback loop. The stochastic synchronization of instantaneously coupled neurons under the influence of white noise can be deliberately controlled by local time-delayed feedback. By appropriate choice of the delay time, synchronization can be either enhanced or suppressed. In delay-coupled neurons, antiphase oscillations can be induced for sufficiently large delay and coupling strength. The additional application of time-delayed self-feedback leads to complex scenarios of synchronized in-phase or antiphase oscillations, bursting patterns or amplitude death. PMID- 19218153 TI - Delays and weakly coupled neuronal oscillators. AB - We use weakly coupled oscillator theory to study the effects of delays on coupled systems of neuronal oscillators. We explore, first, simple pairs with constant delays and then examine the role of distributed delays as would occur in systems with dendritic branches or in networks where there is a distance-dependent conductance delay. In the latter, we use mean field theory to show the emergence of travelling waves and the loss of synchronization. Next, we consider phase models with stronger coupling and delays in the state variables. We show that they have a richer dynamics but one that is still similar to the weakly coupled case. PMID- 19218154 TI - Delays in activity-based neural networks. AB - In this paper, we study the effect of two distinct discrete delays on the dynamics of a Wilson-Cowan neural network. This activity-based model describes the dynamics of synaptically interacting excitatory and inhibitory neuronal populations. We discuss the interpretation of the delays in the language of neurobiology and show how they can contribute to the generation of network rhythms. First, we focus on the use of linear stability theory to show how to destabilize a fixed point, leading to the onset of oscillatory behaviour. Next, we show for the choice of a Heaviside nonlinearity for the firing rate that such emergent oscillations can be either synchronous or anti-synchronous, depending on whether inhibition or excitation dominates the network architecture. To probe the behaviour of smooth (sigmoidal) nonlinear firing rates, we use a mixture of numerical bifurcation analysis and direct simulations, and uncover parameter windows that support chaotic behaviour. Finally, we comment on the role of delays in the generation of bursting oscillations, and discuss natural extensions of the work in this paper. PMID- 19218155 TI - Neural field dynamics with local and global connectivity and time delay. AB - Spatially continuous networks with heterogeneous connections are ubiquitous in biological systems, in particular neural systems. To understand the mutual effects of locally homogeneous and globally heterogeneous connectivity, we investigate the stability of the rest-state activity of a neural field as a function of its connectivity. The variation of the connectivity is operationalized through manipulation of a heterogeneous two-point connection embedded into the otherwise homogeneous connectivity matrix, as well as by variation of connectivity strength and a finite transmission speed. The latter results in a time delay of communication among individual brain areas. We demonstrate that the local connectivity generates the well-known power-law behaviour of the electroencephalographic power spectrum with an exponent close to -2, whereas the global connections generate a more characteristic line spectrum. These spectral characteristics are routinely observed in large-scale topographies of the human brain. PMID- 19218156 TI - Onset of polyspike complexes in a mean-field model of human electroencephalography and its application to absence epilepsy. AB - In this paper, we introduce a modification of a mean-field model used to describe the brain's electrical activity as recorded via electroencephalography (EEG). The focus of the present study is to understand the mechanisms giving rise to the dynamics observed during absence epilepsy, one of the classical generalized syndromes. A systematic study of the data from a number of different subjects with absence epilepsy demonstrates a wide variety of dynamical phenomena in the recorded EEG. In addition to the classical spike and wave activity, there may be polyspike and wave, wave spike or even no discernible spike-wave onset during seizure events. The model we introduce is able to capture all of these different phenomena and we describe the bifurcations giving rise to these different types of seizure activity. We argue that such a model may provide a useful clinical tool for classifying different subclasses of absence epilepsy. PMID- 19218157 TI - Effects of neuromuscular lags on controlling contact transitions. AB - We present a numerical exploration of contact transitions with the fingertip. When picking up objects our fingertips must make contact at specific locations, and-upon contact-maintain posture while producing well-directed force vectors. However, the joint torques for moving the fingertip towards a surface (tau(m)) are different from those for producing static force vectors (tau(f)). We previously described the neural control of such abrupt transitions in humans, and found that unavoidable errors arise because sensorimotor time delays and lags prevent an instantaneous switch between different torques. Here, we use numerical optimization on a finger model to reveal physical bounds for controlling such rapid contact transitions. Resembling human data, it is necessary to anticipatorily switch joint torques to tau(f )at about 30 ms before contact to minimize the initial misdirection of the fingertip force vector. This anticipatory strategy arises in our deterministic model from neuromuscular lags, and not from optimizing for robustness to noise/uncertainties. Importantly, the optimal solution also leads to a trade-off between the speed of force magnitude increase versus the accuracy of initial force direction. This is an alternative to prevailing theories that propose multiplicative noise in muscles as the driver of speed-accuracy trade-offs. We instead find that the speed-accuracy trade-off arises solely from neuromuscular lags. Finally, because our model intentionally uses idealized assumptions, its agreement with human data suggests that the biological system is controlled in a way that approaches the physical boundaries of performance. PMID- 19218158 TI - Balancing with positive feedback: the case for discontinuous control. AB - Experimental observations indicate that positive feedback plays an important role for maintaining human balance in the upright position. This observation is used to motivate an investigation of a simple switch-like controller for postural sway in which corrective movements are made only when the vertical displacement angle exceeds a certain threshold. This mechanism is shown to be consistent with the experimentally observed variations in the two-point correlation for human postural sway. Analysis of first-passage times for this model suggests that this control strategy may slow escape by taking advantage of two intrinsic properties of a stochastic unstable first-order delay differential equation: (i) time delay and (ii) the possibility that the dynamics can be 'temporarily confined' near the origin. PMID- 19218159 TI - Delay effects in the human sensory system during balancing. AB - Mechanical models of human self-balancing often use the Newtonian equations of inverted pendula. While these mathematical models are precise enough on the mechanical side, the ways humans balance themselves are still quite unexplored on the control side. Time delays in the sensory and motoric neural pathways give essential limitations to the stabilization of the human body as a multiple inverted pendulum. The sensory systems supporting each other provide the necessary signals for these control tasks; but the more complicated the system is, the larger delay is introduced. Human ageing as well as our actual physical and mental state affects the time delays in the neural system, and the mechanical structure of the human body also changes in a large range during our lives. The human balancing organ, the labyrinth, and the vision system essentially adapted to these relatively large time delays and parameter regions occurring during balancing. The analytical study of the simplified large-scale time-delayed models of balancing provides a Newtonian insight into the functioning of these organs that may also serve as a basis to support theories and hypotheses on balancing and vision. PMID- 19218161 TI - [Change in surgical treatment of esophageal cancer in the course of the past 50 years]. AB - Oesophageal surgery--apart from sporadic attempts--has a history of about fifty years. It was traditionally fallen into the realm of thoracic surgery developing collaterally and accomplished by the development of anaesthesiology and perioperative care. Initial surgery of the oesophagus begun by procedures aimed at tumours of the lower third and those of the gastroesophageal junction and the cardia. Surgical procedures for esophageal cancer became widespread by 1970's, leading to establishment of oesophageal surgical centres. Partial resections were succeeded by subtotal resections by 1980's. Hypopharyngeal and cervical oesophageal tumours were routinely extirpated in specialized centres by the 1990's. Extended lymph node dissection became routine and generally accepted. By the end of the decade, the importance of neoadjuvant radio-chemotherapy was highlighted and became inevitable. Growing experience of open transthoracic and blunt transhiatal resections without thoracotomy led to the onset of early thoracoscopic and laparoscopic procedures. The current practice for intraepithelial neoplasms is a minimally invasive procedure, such as endoscopic mucosectomy beside blunt transhiatal resection without thoracotomy. In case of submucosal tumours transthoracic or transhiatal blunt subtotal resections are recommended with 2-field lymphadenectomy. Solely subtotal resection with 2- or 3 field lymphadenectomy can be considered as curative intervention for advanced stage T2 cancer. In cases of T3 and T4 mid, or upper third and cervical neoplasms neoadjuvant radio-chemotherapy is recommended. Curative resection is only considered for responders. PMID- 19218162 TI - [Evolution of endocrine surgery: past, present and future]. PMID- 19218163 TI - [Dr. Pal Ondrejka, director of the 2nd Department of Surgery of Semmelweis University - 60th birthday]. PMID- 19218164 TI - [Prognostic role of vascularisation and proliferation in rectal cancer with liver metastasis]. AB - BACKGROUND: The present study was designed to provide an analysis of factors for angiogenesis and proliferation. MATERIAL AND METHOD: We analyzed tumor tissues from 37 rectal cancer patients with concurrent or subsequent liver metastasis underwent preoperative radiotherapy, surgery and adjuvant chemotherapy. Immunohistochemistry was used for expression of proliferation (staining with anti Ki67: MIB-1) and for detection of microvessel density (MVD, expressed by CD34). Clinicopathological findings were compared with outcome with emphasis to IHC. RESULTS: A vascular enumeration and pN status and the time of presence of the metastases has shown prognostic role along with the factors above. Increased proliferative activity of the tumor as expressed by MIB-1 staining has no prognostic value, similarly to the localization of tumor, gender, age or grading. SUMMARY: Different prognostic and predictive factors in colorectal cancer have been reported. Higher pN status and tumor vascularisation has been linked to poor prognosis in overall survival and tumor recurrence. PMID- 19218165 TI - [Precursors of Pancreatic cancer: Intraepithelial Neoplasia (PanIN) and Intraductal Papillary Mucinous Neoplasms (IPMN)]. AB - Pancreatic Intraepithelial Neoplasia (PanIN1-3) and Intraductal Papillary Mucinous Neoplasms (IPMN) putative precursors of and associated to pancreatic cancer represent a distinct, however pathologically heterogenous entity. Recently a new classification and nomenclature was established. We review the clinicopathologic and cytogenetic characteristics of PanIN and IPMN highlighting the nonaggressive biological behavior of intraepithelial and intraductal neoplasms which, in part explain their favorable prognosis. PMID- 19218166 TI - [An uncommon malignancy of the breast: dermatofibrosarcoma protuberans with fibrosarcomatous transformation (DFSP-FS)]. AB - Dermatofibrosarcoma protuberans (DFSP) is a morphologically distinct dermal/subcutaneous fibrohystiocytic neoplasm seen mainly on the trunk and the proximal extremities, which generally is regarded as a low-grade or "borderline" neoplasm. Dermatofibrosarcoma protuberans is characterized clinically by locally aggressive growth and a high rate of local recurrence, but distant metastases and tumor-related deaths are very rare. DFSP in the breast is extremely uncommon. Immunohistochemically, DFSP is characterized by positive staining for vimentin and CD34. We present a case of a 65 years old female patient with a giant DFSP developed in the sight of several excised fibroadenoma and neurofibroma treated by radical mastectomy and axillary block-dissection. Perioperative hystological and immunohistochemical images demonstrate the characteristics of the giant nonphylloid sarcoma. PMID- 19218167 TI - [Recurrent retroperitoneal liposarcoma presenting as an incarcerated femoral hernia]. AB - 56-year-old man was admitted to our hospital 18 month after extirpation of retroperitoneal liposarcoma. We diagnosed recurrent tumor in the inguinal fossa. The tumor situated along the femoral artery and propagated to the thigh among the muscles, mimicking incarcerated femoral hernia. We performed resection with synchronous abdominal and femoral exploration. By our knowledge this is the first case report about recurrent retroperitoneal liposarcoma presenting as an incarcerated femoral hernia. PMID- 19218168 TI - [Aortoenteric fistula to the sigmoid colon complicated with entero-grafto cutaneous fistula]. AB - We report a case of a 62-year-old man who presented to the emergency department with acute rectal bleeding. The patient had previous aortoiliac surgery with the utilization of an aorto-bifemoral vascular graft. Diagnosis of secondary aortoenteric fistula was made between the aortoiliac graft and sigmoid colon. This fistula had an entero-cutaneous component. After exploratory laparotomy resection of the sigma, extirpation of the entero-cutaneous fistula, excision of the graft, oversewing of the aortic stump, and extra-anatomical crossover bypass were successfully performed. This study reports a rare type of secondary aortoenteric fistula to the sigmoid colon complicated with an entero-grafto cutaneous component and it describes an unusual and successful surgical treatment. PMID- 19218169 TI - [Remembering Mano Herczel (1862-1918)]. PMID- 19218170 TI - [To the Editor-in-Chief: Remembering Dr. Gabor Petri]. PMID- 19218172 TI - Association between HIV-1 RNA level and CD4 cell count among untreated HIV infected individuals. AB - OBJECTIVES: We examined the significance of plasma HIV-1 RNA levels (or viral load alone) in predicting CD4 cell decline in untreated HIV-infected individuals. METHODS: Data were obtained from the British Columbia Centre for Excellence in HIV/AIDS. Participants included all residents who ever had a viral load determination in the province and who had never taken antiretroviral drugs (N = 890). We analyzed a total of 2074 viral load measurements and 2332 CD4 cell counts. Linear mixed-effects models were used to predict CD4 cell decline over time. RESULTS: Longitudinal viral load was strongly associated with CD4 cell decline over time; an average of 1 log(10) increase in viral load was associated with a 55-cell/mm(3) decrease in CD4 cell count. CONCLUSIONS: Our results support the combined use of CD4 cell count and viral load as prognostic markers in HIV infected individuals before the introduction of antiretroviral therapy. PMID- 19218173 TI - Sexually transmitted diseases among adults who had been abused and neglected as children: a 30-year prospective study. AB - OBJECTIVES: We examined associations between childhood abuse and neglect and the risk in adulthood for sexually transmitted diseases. METHODS: In a prospective cohort design, we matched children aged 0 to 11 years with documented cases of abuse or neglect during 1967 to 1971 with a control group of children who had not been maltreated (754 participants in all) and followed them into adulthood. Information about lifetime history of sexually transmitted diseases was collected as part of a medical status examination when participants were approximately 41 years old. RESULTS: Childhood sexual abuse increased risk for any sexually transmitted disease (odds ratio [OR] = 1.94; 95% confidence interval [CI] = 1.00, 3.77; P = .05) and more than 1 type of sexually transmitted disease (OR = 3.33; 95% CI = 1.33, 8.22; P = .01). Physical abuse increased risk for more than 1 type of sexually transmitted disease (OR = 3.61; 95% CI = 1.39, 9.38; P = .009). CONCLUSIONS: Our results provided the first prospective evidence that child physical and sexual abuse increases risk for sexually transmitted diseases. Early screening and interventions are needed to identify and prevent sexually transmitted diseases among child abuse victims. PMID- 19218174 TI - HIV risk associated with gay bathhouses and sex clubs: findings from 2 seattle surveys of factors related to HIV and sexually transmitted infections. AB - OBJECTIVES: We studied the HIV risk behaviors of patrons of the 3 commercial sex venues for men in Seattle, Washington. METHODS: We conducted cross-sectional, observational surveys in 2004 and 2006 by use of time-venue cluster sampling with probability proportional to size. Surveys were anonymous and self-reported. We analyzed the 2004 data to identify patron characteristics and predictors of risk behaviors and compared the 2 survey populations. RESULTS: Fourteen percent of respondents reported a previous HIV-positive test, 14% reported unprotected anal intercourse, and 9% reported unprotected anal intercourse with a partner of unknown or discordant HIV status during the current commercial sex venue visit. By logistic regression, recent unprotected anal intercourse outside of a commercial sex venue was independently associated with unprotected anal intercourse. Sex venue site and patron drug use were strongly associated with unprotected anal intercourse at the crude level. The 2004 and 2006 survey populations did not differ significantly in demographics or behaviors. CONCLUSIONS: Patron and venue-specific characteristics factors may each influence the frequency of HIV risk behaviors in commercial sex venues. Future research should evaluate the effect of structural and individual-level interventions on HIV transmission. PMID- 19218175 TI - Contribution of race/ethnicity and country of origin to variations in lifetime reported asthma: evidence for a nativity advantage. AB - OBJECTIVES: We assessed the relative contribution of Hispanic ethnicity, country of origin, and nativity to lifetime prevalence of asthma among mothers and children enrolled in the Project on Human Development in Chicago Neighborhoods. METHODS: We used multilevel models to analyze data from wave 3 of the Project on Human Development in Chicago Neighborhoods study (2000 to 2001). Mothers reported physician-diagnosed asthma for themselves and their children. Maternal race, ethnicity, country of origin, and nativity were the predictors of interest. RESULTS: We found substantial heterogeneity in lifetime asthma within Hispanic subgroups for mothers and children. Hispanics of non-Mexican origin had greater odds of having asthma than did non-Hispanic Whites; respondents of Mexican origin did not differ from non-Hispanic Whites. Odds of experiencing asthma were more strongly related to nativity than to race, Hispanic ethnicity, or country of origin. Only immigrant Mexicans reported asthma prevalence lower than that of native non-Hispanic Whites. CONCLUSIONS: Nativity is a strong predictor of lifetime asthma prevalence, suggesting the importance of potential interactions between genetic susceptibilities and environmental factors in both the native and the host countries. PMID- 19218176 TI - Health care access and sexually transmitted infection screening frequency among at-risk Massachusetts men who have sex with men. AB - OBJECTIVES: We sought to assess risk exposures, health care access, and screening rates for HIV and sexually transmitted infections (STIs) among men who have sex with men (MSM) in Massachusetts. METHODS: We used a modified respondent-driven sampling method to collect data between March 2006 and May 2007. Overall, 126 MSM completed a survey. RESULTS: Seventy percent of participants reported unprotected receptive anal intercourse with at least 1 nonmonogamous male partner; 50% reported having had a previous STI. Although 98% had visited a health care provider in the previous year, 39% had not been screened for STIs during the previous 2 years. Bisexual respondents were less likely to have told their health care providers that they engage in male-to-male sexual contact (OR = 4.66; P < .001), less likely to have been tested for STIs during in the previous 2 years (OR = 6.91; P < .001), and more likely to engage in insertive anal intercourse without a condom with an HIV-infected partner (OR = 5.04; P < .005) than were non bisexual respondents. CONCLUSIONS: Clinicians need to assess sexual risk-taking behaviors and more routinely screen for STIs among sexually active men regardless of disclosure of a history of having sex with men. PMID- 19218177 TI - Black men who have sex with men and the association of down-low identity with HIV risk behavior. AB - Black men "on the down low" have been considered prime agents of HIV transmission in the Black community despite little empirical evidence. We assessed the relationship between down-low identification and sexual risk outcomes among 1151 Black MSM. Down-low Identification was not associated with unprotected anal or vaginal sex with male or female partners. Future HIV prevention programs and research should target sexual risk behaviors of Black men, irrespective of identity, and not focus on the "down low." PMID- 19218178 TI - Condom use and high-risk sexual acts in adult films: a comparison of heterosexual and homosexual films. AB - OBJECTIVES: We compared the prevalence of condom use during a variety of sexual acts portrayed in adult films produced for heterosexual and homosexual audiences to assess compliance with state Occupational Health and Safety Administration regulations. METHODS: We analyzed 50 heterosexual and 50 male homosexual films released between August 1, 2005, and July 31, 2006, randomly selected from the distributor of 85% of the heterosexual adult films released each year in the United States. RESULTS: Penile-vaginal intercourse was protected with condoms in 3% of heterosexual scenes. Penile-anal intercourse, common in both heterosexual (42%) and homosexual (80%) scenes, was much less likely to be protected with condoms in heterosexual than in homosexual scenes (10% vs 78%; P < .001). No penile-oral acts were protected with condoms in any of the selected films. CONCLUSIONS: Heterosexual films were much less likely than were homosexual films to portray condom use, raising concerns about transmission of HIV and other sexually transmitted diseases, especially among performers in heterosexual adult films. In addition, the adult film industry, especially the heterosexual industry, is not adhering to state occupational safety regulations. PMID- 19218179 TI - Effects of a hepatitis C virus educational intervention or a motivational intervention on alcohol use, injection drug use, and sexual risk behaviors among injection drug users. AB - OBJECTIVES: We compared the effects of 2 interventions on alcohol use, use of a new syringe at last injection, and condom use at last sexual encounter in a community sample of injection drug users. METHODS: Between 2003 and 2006, 851 out of-treatment injection drug users were recruited in Raleigh, NC, and Durham, NC, through street outreach and were randomly assigned to either a 6-session educational intervention or a 6-session motivational intervention. Intervention effects were examined at 6 and 12 months after enrollment. RESULTS: In multiple logistic regression analyses adjusted for baseline alcohol use and HCV status, participants assigned to the motivational intervention were significantly less likely than were participants in the educational intervention to be drinking at the 6-month follow-up (odds ratio = 0.67; 95% confidence interval = 0.46, 0.97). There were no significant between-group differences in use of a new syringe at last injection or condom use at last sexual encounter at either follow-up. CONCLUSIONS: Reducing alcohol use among persons with HCV may slow disease progression and provide important health benefits. Additional strategies are needed for slowing HCV disease progression until more effective HCV treatments are available. PMID- 19218180 TI - Evaluation of the impact of news coverage of an HIV multiclass drug-resistant cluster in Seattle, Washington. AB - OBJECTIVES: In February 2007, Public Health-Seattle and King County issued a press release describing a cluster of multiclass drug-resistant HIV cases among men who had sex with men (MSM). We evaluated the effect of the press release among MSM in the Seattle area. METHODS: We administered a rapid assessment survey at venues where MSM congregate. Eligible participants were men who had sex with men in the past year, were older than 18 years, and were residents of western Washington State. RESULTS: Among 325 participants, 57% heard or saw messages related to the press release. Of these, 87% remembered 1 or more key points, but only 5% remembered key prevention messages. Ninety-eight percent of participants thought it was important for the health department to get the message out about drug-resistant HIV. CONCLUSIONS: The press release was found to be a useful and well-received method to inform the public about an HIV drug-resistant cluster. Low retention and nonprominent coverage of key prevention messages suggests that health departments using press releases as a prevention tool need to carefully consider placement and emphasis of those messages in a press statement. PMID- 19218181 TI - Risky sexual behavior, bleeding caused by intimate partner violence, and hepatitis C virus infection in patients of a sexually transmitted disease clinic. AB - OBJECTIVES: We sought to investigate independent contributions of risky sexual behaviors and bleeding caused by intimate partner violence to prediction of HCV infection. METHODS: We conducted a case-control study of risk factors among patients of a sexually transmitted disease clinic with and without HCV antibodies, group-matched by age. RESULTS: Multivariate analyses indicated that Black race (odds ratio [OR] = 2.4; 95% confidence interval [CI] = 1.3, 4.4), injection drug use (OR = 20.3; 95% CI = 10.8, 37.8), sharing straws to snort drugs (OR = 1.8; 95% CI = 1.01, 3.0), sharing razors (OR = 7.8; 95% CI = 2.0, 31.0), and exposure to bleeding caused by intimate partner violence (OR = 5.5; 95% CI = 1.4, 22.8) contributed significantly to the prediction of HCV infection; risky sexual behavior and exposure to blood or sores during sexual intercourse did not. CONCLUSIONS: HCV risk among patients of a sexually transmitted disease clinic can be explained by direct blood exposure, primarily through injection drug use. Exposure to bleeding caused by intimate partner violence may be a previously unrecognized mechanism for HCV transmission associated with risky sexual behavior. PMID- 19218182 TI - Racial discrimination and racial identity attitudes in relation to self-rated health and physical pain and impairment among two-spirit American Indians/Alaska Natives. AB - OBJECTIVES: We examined associations between racial discrimination and actualization, defined as the degree of positive integration between self identity and racial group identity, and self-rated health and physical pain and impairment. METHODS: We used logistic regressions to analyze data from 447 gay, lesbian, bisexual, and other sexual-minority American Indians/Alaska Natives. RESULTS: Greater self-reported discrimination was associated with higher odds of physical pain and impairment (odds ratio [OR] = 1.42; 95% confidence interval [CI] = 1.13, 1.78); high levels of actualization were associated with lower odds of physical pain and impairment (OR = 0.59; 95% CI = 0.35, 0.99) and self-rated fair or poor health (OR = 0.54; 95% CI = 0.32, 0.90). Actualization also moderated the influence of discrimination on self-rated health (t = -2.33; P = .020). Discrimination was positively associated with fair or poor health among participants with low levels of actualization, but this association was weak among those with high levels of actualization. CONCLUSIONS: Among two-spirit American Indians/Alaska Natives, discrimination may be a risk factor for physical pain and impairment and for fair or poor self-rated health among those with low levels of actualization. Actualization may protect against physical pain and impairment and poor self-rated health and buffer the negative influence of discrimination. PMID- 19218183 TI - Demographic characteristics and survival with AIDS: health disparities in Chicago, 1993-2001. AB - OBJECTIVES: We examined correlations between survival and race/ethnicity, age, and gender among persons who died from AIDS-related causes. METHODS: We estimated survival among 11 022 persons at 12, 36, and 60 months after diagnosis with AIDS in 1993 through 2001 and reported through 2003 to the Chicago Department of Public Health. We estimated hazard ratios (HRs) by demographic and risk characteristics. RESULTS: All demographic groups had higher 5-year survival rates after the introduction of highly active retroviral therapy (1996-2001) than before (1993-1995). The HR for non-Hispanic Blacks to Whites was 1.18 in 1993 to 1995 and 1.51 (P < .01) in 1996 to 2001. The HR for persons 50 years or older to those younger than 30 years was 1.63 in 1993-1995 and 2.28 (P < .01) in 1996 2001. The female-to-male HR was 0.90 in 1993-1995 and 1.20 (P < .02) in 1996 2001. CONCLUSIONS: The risk of death was higher for non-Hispanic Blacks and Hispanics than for non-Hispanic Whites. Interventions are needed to increase early access to care for disadvantaged groups. PMID- 19218184 TI - HIV prevention technology transfer: challenges and strategies in the real world. AB - OBJECTIVES: We examined implementation of evidence-based interventions for HIV prevention at community-based organizations in Los Angeles County, CA. METHODS: We conducted 2 waves of interviews with 34 organization staff members. We analyzed activities reported by staff in the phases (preimplementation, implementation, and maintenance and evolution) and activities defined by the technology transfer model for evidence-based HIV prevention interventions. RESULTS: Staff members were able to select, adapt, and implement evidence-based HIV prevention interventions despite challenges in each phase of technology transfer. Preimplementation challenges included lack of information and poor fit between the interventions and organizations' clients. Implementation challenges included retention of participants across intervention sessions and staff turnover. A challenge in the maintenance and evolution phase was enhancing staff skills in outcome monitoring and cost analyses. CONCLUSIONS: Technical assistance must be matched to the specific challenges found in each phase of technology transfer. Successful transfer of evidence-based HIV prevention interventions will depend on their continued uptake and use by organization staff. This study highlights directions for improving communications regarding appropriate modifications to these interventions and for organizational planning to continue adapted interventions. PMID- 19218185 TI - A brief, clinic-based, safer sex intervention for heterosexual African American men newly diagnosed with an STD: a randomized controlled trial. AB - OBJECTIVE: We evaluated the efficacy of a brief, clinic-based, safer sex program administered by a lay health adviser for young heterosexual African American men newly diagnosed with a sexually transmitted disease (STD). METHODS: Subsequent to STD diagnosis, eligible men (N = 266; aged 18-29 years) were randomized to either a personalized, single-session intervention (delivered by a lay health adviser) or standard of care. We conducted behavioral assessments at baseline and 3 months postintervention (retention was 74.1%). We also conducted a 6-month clinic record review. RESULTS: Compared to men randomized to the control condition, those receiving the intervention were significantly less likely to acquire subsequent STDs (50.4% vs 31.9%; P = .002) and more likely to report using condoms during last sexual intercourse (72.4% vs 53.9%; P = .008). They also reported fewer sexual partners (mean 2.06 vs 4.15; P < .001) and fewer acts of unprotected sex (mean 12.3 vs 29.4; P = .045). Based on a 9-point rating scale, men in the intervention group had higher proficiency scores for condom application skills (mean difference = 3.17; P < .001). CONCLUSION: A brief clinic-based intervention delivered by a lay health adviser may be an efficacious strategy to reduce incident STDs among young heterosexual African American men. PMID- 19218187 TI - Molecular phenotyping of mannosyltransferases-deficient Candida albicans cells by high-resolution magic angle spinning NMR. AB - The yeast Candida albicans is an opportunistic pathogen that causes infections in immunocompromised individuals with a high morbidity and mortality levels. Recognition of yeasts by host cells is directly mediated by cell wall components of the yeast, including a wide range of abundantly expressed glycoconjugates. Of particular interest in C. albicans are the beta-mannosylated epitopes that show a complex expression pattern on N-glycan moiety of phosphopeptidomannans and are absent in the non-pathogenic species Saccharomyces cerevisiae. Being known as potent antigens for the adaptive immune response and elicitors of specific infection-protective antibodies, the exact delineation of beta-mannosides regulation and expression pathways has lately become a major milestone toward the comprehension of host-pathogen interplay. Using the newly developed HR-MAS NMR methodology, we demonstrate the possibility of assessing the general profiles of cell-surface-exposed glycoconjugates from intact living yeast cells without any prior purification step. This technique permitted to directly observe structural modifications of surface expressed phosphodiester-linked beta-mannosides on a series of deletion strains in beta-mannosyltransferases and phospho mannosyltransferases compared with their parental strains. PMID- 19218186 TI - Perceived everyday racism, residential segregation, and HIV testing among patients at a sexually transmitted disease clinic. AB - OBJECTIVES: More than one quarter of HIV-infected people are undiagnosed and therefore unaware of their HIV-positive status. Blacks are disproportionately infected. Although perceived racism influences their attitudes toward HIV prevention, how racism influences their behaviors is unknown. We sought to determine whether perceiving everyday racism and racial segregation influence Black HIV testing behavior. METHODS: This was a clinic-based, multilevel study in a North Carolina city. Eligibility was limited to Blacks (N = 373) seeking sexually transmitted disease diagnosis or screening. We collected survey data, block group characteristics, and lab-confirmed HIV testing behavior. We estimated associations using logistic regression with generalized estimating equations. RESULTS: More than 90% of the sample perceived racism, which was associated with higher odds of HIV testing (odds ratio = 1.64; 95% confidence interval = 1.07, 2.52), after control for residential segregation, and other covariates. Neither patient satisfaction nor mechanisms for coping with stress explained the association. CONCLUSIONS: Perceiving everyday racism is not inherently detrimental. Perceived racism may improve odds of early detection of HIV infection in this high-risk population. How segregation influences HIV testing behavior warrants further research. PMID- 19218188 TI - Cooperative binding of L-trp to human tryptophan 2,3-dioxygenase: resonance Raman spectroscopic analysis. AB - Tryptophan 2,3-dioxygenase (TDO) is a tetrameric enzyme that catalyses the oxidative cleavage of l-tryptophan (l-Trp) to N-formylkynurenine by the addition of O(2) across the 2,3-bond of the indole ring. This reaction is the first and rate-limiting step in the kynurenine pathway in mammals. In the present study, we measured the conformational changes in the haem pocket of recombinant human TDO (rhTDO) in ferric form that are induced by l-Trp binding using both resonance Raman and optical absorption spectroscopies. The deconvolution analysis of the haem Raman bands at various concentrations of l-Trp revealed that the wild-type enzyme exhibits homotropic cooperativity in l-Trp binding, which was confirmed by a change in the optical absorption spectra. Mutation analysis showed that the Y42F mutant abolished the cooperative binding, and that the H76A mutant considerably reduced the catalytic activity. These data and the inter-subunit contacts reported in the bacterial TDO structure suggest that the Y42 of rhTDO is responsible for the cooperative binding of l-Trp by participating in the active site of the adjacent subunit. PMID- 19218189 TI - Construction of the plasmid, expression by Chinese hamster ovary cell, purification and characterization of the first three short consensus repeat modules of human complement receptor type 1. AB - Short consensus repeat (SCR1-3), the first three SCR modules from N-terminus of type 1 complement receptor (CR1), is expected to accelerate dissociation of complement components and suppress complement activity by binding the main component of complement C4b. In order to clarify the three-dimensional structure, which triggers the activity of SCR1-3 on complement, we constructed an over expression system in CHO DG44 cells which facilitated mass production of SCR1-3. The mass production was achieved by a two-stage culture system and optimum culture conditions using ASF104N medium and MTX-, NaBu-containing alpha-MEM/10% FBS medium, respectively. The constructed gene of SCR1-3 was confirmed by restriction enzyme digestion and DNA sequence analysis, and the expressed protein by CHO DG44 cells was confirmed by western blotting. The expressed SCR1-3 was proved containing N-linked sugar chain, an important factor to the proper expression of protein, by the cleavage with glycosidase of N-linked oligosaccharide (PNGase F). The suppression effect of the yield protein on complement-mediated inflammation was investigated by haemolytic assay and necrosis assay of stromal cells. Both assays showed that SCR1-3 possessed complement control activity. However, residing sugar chain on SCR1-3 did not show significant difference in the complement control activity. PMID- 19218190 TI - Gene identification and characterization of 5-formyl-3-hydroxy-2-methylpyridine 4 carboxylic acid 5-dehydrogenase, an NAD+-dependent dismutase. AB - A chromosomal gene, mlr6793, in Mesorhizobium loti was identified as the gene encoding 5-formyl-3-hydroxy-2-methylpyridine 4-carboxylic acid (FHMPC) dehydrogenase (dismutase) involved in the degradation pathway for pyridoxine (vitamin B(6)). The homogenously purified recombinant enzyme has a molecular mass of 59.1 kDa and is a homodimeric protein. FHMPC dehydrogenase catalyses practically irreversible oxidation (k(cat) = 204 s(-1)) of FHMPC (K(m) = 48.2 microM) by NAD(+) (K(m) = 34.3 microM) to 3-hydroxy-2-methyl-pyridine 4, 5 dicarboxylic acid (HMPDC), and practically irreversible reduction (k(cat) = 217 s(-1)) of FHMPC (K(m) = 24.9 microM) by NADH (K(m) = 12.4 microM) to 4-pyridoxic acid. When the enzyme reaction was started with the combination of FHMPC and NAD(+) or that of FHMPC and NADH, HMPDC and 4-pyridoxic acid were produced in an almost equimolar ratio throughout the reaction. FHMPC dehydrogenase belongs to the 3-hydroxyacyl-CoA dehydrogenase family with 31% identity with the human enzyme: it has probable catalytic diad residues, i.e. His137 and Glu149. The H137L mutant enzyme showed no measurable activity. The E149Q one was stable in contrast to the corresponding human 3-hydroxyacyl-CoA dehydrogenase mutant, and showed unique pH optima depending on the co-substrates used for the reaction. PMID- 19218191 TI - Interleukin-13-induced MUC5AC is regulated by 15-lipoxygenase 1 pathway in human bronchial epithelial cells. AB - RATIONALE: 15-Lipoxygenase-1 (15LO1) and MUC5AC are highly expressed in asthmatic epithelial cells. IL-13 is known to induce 15LO1 and MUC5AC in human airway epithelial cells in vitro. Whether 15LO1 and/or its product 15-HETE modulate MUC5AC expression is unknown. OBJECTIVES: To determine the expression of 15LO1 in freshly harvested epithelial cells from subjects with asthma and normal control subjects and to determine whether IL-13-induced 15LO1 expression and activation regulate MUC5AC expression in human bronchial epithelial cells in vitro. METHODS: Human airway epithelial cells from subjects with asthma and normal subjects were evaluated ex vivo for 15LO1 and MUC5AC expression. The impact of 15LO1 on MUC5AC expression in vitro was analyzed by inhibiting 15LO1 through pharmacologic (PD146176) and siRNA approaches in human bronchial epithelial cells cultured under air-liquid interface. We analyzed 15 hydroxyeicosatetraenoic acid (15-HETE) by liquid chromatography/UV/mass spectrometry. MUC5AC and 15LO1 were analyzed by real-time RT-PCR, immunofluoresence, and Western blot. MEASUREMENTS AND MAIN RESULTS: Epithelial 15LO1 expression increased with asthma severity (P < 0.0001). 15LO1 significantly correlated with MUC5AC ex vivo and in vitro. IL-13 increased 15LO1 expression and stimulated formation of two molecular species of 15-HETE esterified to phosphotidylethanolamine (15-HETE-PE). Inhibition of 15LO1 suppressed 15-HETE-PE and decreased MUC5AC expression in the presence of IL-13 stimulation. The addition of exogenous 15-HETE partially restored MUC5AC expression. CONCLUSIONS: Epithelial 15LO1 expression increases with increasing asthma severity. IL-13 induction of 15-HETE-PE enhances MUC5AC expression in human airway epithelial cells. High levels of 15LO1 activity could contribute to the increases of MUC5AC observed in asthma. PMID- 19218192 TI - Genetic risk factors for portopulmonary hypertension in patients with advanced liver disease. AB - RATIONALE: Portopulmonary hypertension (PPHTN) occurs in 6% of liver transplant candidates. The pathogenesis of this complication of portal hypertension is poorly understood. OBJECTIVES: To identify genetic risk factors for PPHTN in patients with advanced liver disease. METHODS: We performed a multicenter case control study of patients with portal hypertension. Cases had a mean pulmonary artery pressure >25 mm Hg, pulmonary vascular resistance >240 dynes.s(-1).cm(-5), and pulmonary capillary wedge pressure < or =15 mm Hg. Controls had a right ventricular systolic pressure < 40 mm Hg (if estimated) and normal right-sided cardiac morphology by transthoracic echocardiography. We genotyped 1,079 common single nucleotide polymorphisms (SNPs) in 93 candidate genes in each patient. MEASUREMENTS AND MAIN RESULTS: The study sample included 31 cases and 104 controls. Twenty-nine SNPs in 15 candidate genes were associated with the risk of PPHTN (P < 0.05). Multiple SNPs in the genes coding for estrogen receptor 1, aromatase, phosphodiesterase 5, angiopoietin 1, and calcium binding protein A4 were associated with the risk of PPHTN. The biological relevance of one of the aromatase SNPs was supported by an association with plasma estradiol levels. CONCLUSIONS: Genetic variation in estrogen signaling and cell growth regulators is associated with the risk of PPHTN. These biologic pathways may elucidate the mechanism for the development of PPHTN in certain patients with severe liver disease. PMID- 19218193 TI - Uric acid is a danger signal activating NALP3 inflammasome in lung injury inflammation and fibrosis. AB - RATIONALE: Lung injury leads to pulmonary inflammation and fibrosis through myeloid differentiation primary response gene 88 (MyD88) and the IL-1 receptor 1 (IL-1R1) signaling pathway. The molecular mechanisms by which lung injury triggers IL-1beta production, inflammation, and fibrosis remain poorly understood. OBJECTIVES: To determine if lung injury depends on the NALP3 inflammasome and if bleomycin (BLM)-induced lung injury triggers local production of uric acid, thereby activating the NALP3 inflammasome in the lung. METHODS: Inflammation upon BLM administration was evaluated in vivo in inflammasome deficient mice. Pulmonary uric acid accumulation, inflammation, and fibrosis were analyzed in mice treated with the inhibitor of uric acid synthesis or with uricase, which degrades uric acid. MEASUREMENTS AND MAIN RESULTS: Lung injury depends on the NALP3 inflammasome, which is triggered by uric acid locally produced in the lung upon BLM-induced DNA damage and degradation. Reduction of uric acid levels using the inhibitor of uric acid synthesis allopurinol or uricase leads to a decrease in BLM-induced IL-1beta production, lung inflammation, repair, and fibrosis. Local administration of exogenous uric acid crystals recapitulates lung inflammation and repair, which depend on the NALP3 inflammasome, MyD88, and IL-1R1 pathways and Toll-like receptor (TLR)2 and TLR4 for optimal inflammation but are independent of the IL-18 receptor. CONCLUSIONS: Uric acid released from injured cells constitutes a major endogenous danger signal that activates the NALP3 inflammasome, leading to IL-1beta production. Reducing uric acid tissue levels represents a novel therapeutic approach to control IL-1beta production and chronic inflammatory lung pathology. PMID- 19218194 TI - Lymphoid follicle cells in chronic obstructive pulmonary disease overexpress the chemokine receptor CXCR3. AB - RATIONALE: The mechanisms underlying formation of lung lymphoid follicles (LF) in chronic obstructive pulmonary disease (COPD) are unknown. The chemokine receptor CXCR3 regulates immune responses in secondary lymphoid structures elsewhere in the body and is highly expressed by Th1 lymphocytes in the airway in COPD. Because chemokine receptors control inflammatory cell homing to inflamed tissue, we reasoned that CXCR3 may contribute to LF formation in COPD. OBJECTIVES: We assessed the expression of CXCR3 and its ligands (IP-10/CXCL10, Mig/CXCL9, and ITAC/CXCL11) by LF cells in never-smokers, smokers without COPD, and subjects with COPD. METHODS: CXCR3, IP-10, Mig, and ITAC expression were assessed in lung sections from 46 subjects (never-smokers, smokers without COPD [S], and subjects with COPD in GOLD stages 1-4) by immunohistochemistry. MEASUREMENTS AND MAIN RESULTS: CXCR3-expressing T cells (CD8+ or CD4+) and B cells (CD20+) were topographically distributed at the follicle periphery and center, respectively. The percentage of immunohistochemically identified CXCR3+ cells increased progressively while proceeding from S through GOLD 3-4 (P < 0.01 for GOLD 3-4 vs. S). Moreover, the number of CXCR3+ follicular cells correlated inversely with FEV(1) (r = 0.60). The CXCR3 ligands IP-10 and Mig were expressed by several cell types in and around the follicle, including CD68+ dendritic cells/ macrophages, airway epithelial cells, endothelial cells, and T and B cells. CONCLUSIONS: These results suggest that LF form in the COPD lung by recruitment and/or retention of CXCR3-expressing T and B lymphocytes, which are attracted to the region through production of CXCR3 ligands IP-10 and Mig by lung structural and follicular cells. PMID- 19218195 TI - Role of lung-marginated monocytes in an in vivo mouse model of ventilator-induced lung injury. AB - RATIONALE: Recruited leukocytes play an important role in ventilator-induced lung injury, although studies have focused predominantly on neutrophils. Inflammatory subset Gr-1(high) monocytes are recruited to sites of inflammation and have been implicated in acute lung injury induced by systemic endotoxin. OBJECTIVES: To investigate the recruitment and role of Gr-1(high) monocytes in an in vivo mouse model of ventilator-induced lung injury. METHODS: Anesthetized mice were ventilated with low or high stretch. Flow cytometry was used to quantify monocyte subset margination to the lungs, and to assess their in situ cellular activation in response to mechanical stretch. To investigate monocyte involvement in lung injury progression, a two-hit model was used, with a subclinical dose of lipopolysaccharide (intraperitoneal) given 2 hours prior to high-stretch ventilation. In some animals, monocytes were depleted using intravenous clodronate liposomes. Development of lung injury was assessed in ventilated animals by peak inspiratory pressure and respiratory system mechanics. MEASUREMENTS AND MAIN RESULTS: High-stretch ventilation induced significant pulmonary margination of Gr-1(high) but not Gr-1(low) monocytes compared with nonventilated mice. These monocytes displayed increased activation status, with higher CD11b (vs. nonventilated mice) and lower L-selectin expression (vs. low stretch ventilation). Lipopolysaccharide challenge led to enhanced lung margination of Gr-1(high) monocytes and neutrophils, and sensitized the lungs to high stretch-induced pulmonary edema. Clodronate-liposome pretreatment depleted lung monocytes (but not neutrophils) and significantly attenuated lung injury. CONCLUSIONS: High-stretch mechanical ventilation promotes pulmonary margination of activated Gr-1(high) monocytes, which play a role in the progression of ventilator-induced lung injury. PMID- 19218197 TI - Cryo-electron tomography of porcine reproductive and respiratory syndrome virus: organization of the nucleocapsid. AB - Porcine reproductive and respiratory virus (PRRSV) is an enveloped positive-sense RNA virus of the family Arteriviridae that causes severe and persistent disease in pigs worldwide. The PRRSV virion consists of a lipid envelope that contains several envelope proteins surrounding a nucleocapsid core that encapsidates the RNA genome. To provide a better understanding of the structure and assembly of PRRSV, we have carried out cryo-electron microscopy and tomographic reconstruction of virions grown in MARC-145 cells. The virions are pleomorphic, round to egg-shaped particles with an average diameter of 58 nm. The particles display a smooth outer surface with only a few protruding features, presumably corresponding to the envelope protein complexes. The virions contain a double layered, hollow core with an average diameter of 39 nm, which is separated from the envelope by a 2-3 nm gap. Analysis of the three-dimensional structure suggests that the core is composed of a double-layered chain of nucleocapsid proteins bundled into a hollow ball. PMID- 19218196 TI - Gene expression profiling identifies MMP-12 and ADAMDEC1 as potential pathogenic mediators of pulmonary sarcoidosis. AB - RATIONALE: Little is known about the genetic regulation of granulomatous inflammation in sarcoidosis. OBJECTIVES: To determine if tissue gene array analysis would identify novel genes engaged in inflammation and lung remodeling in patients with sarcoidosis. METHODS: Gene expression analysis was performed on tissues obtained from patients with sarcoidosis at the time of diagnosis (untreated) (n = 6) compared with normal lung tissue (n = 6). Expression of select genes was further confirmed in lung tissue from a second series of patients with sarcoidosis and disease-free control subjects (n = 11 per group) by semi-quantitative RT-PCR. Interactive gene networks were identified in patients with sarcoidosis using Ingenuity Pathway Analysis (Ingenuity Systems, Inc., Redwood, CA) software. The expression of proteins corresponding to selected overexpressed genes was determined using fluorokine multiplex analysis, and immunohistochemistry. Selected genes and proteins were then analyzed in bronchoalveolar lavage fluid in an independent series of patients with sarcoidosis (n = 36) and control subjects (n = 12). MEASUREMENTS AND MAIN RESULTS: A gene network engaged in Th1-type responses was most significantly overexpressed in the sarcoidosis lung tissues, including genes not previously reported in the context of sarcoidosis (e.g., IL-7). MMP-12 and ADAMDEC1 transcripts were most highly expressed (> 25-fold) in sarcoidosis lung tissues, corresponding with increased protein expression by immunohistochemistry. MMP-12 and ADAMDEC1 gene and protein expression were increased in bronchoalveolar lavage samples from patients with sarcoidosis, correlating with disease severity. CONCLUSIONS: Tissue gene expression analyses provide novel insights into the pathogenesis of pulmonary sarcoidosis. MMP-12 and ADAMDEC1 emerge as likely mediators of lung damage and/or remodeling and may serve as markers of disease activity. PMID- 19218198 TI - TRIM22 E3 ubiquitin ligase activity is required to mediate antiviral activity against encephalomyocarditis virus. AB - The interferon (IFN) system is a major effector of the innate immunity that allows time for the subsequent establishment of an adaptive immune response against a wide-range of pathogens. Their diverse biological actions are thought to be mediated by the products of specific but usually overlapping sets of cellular genes induced in the target cells. Ubiquitin ligase members of the tripartite motif (TRIM) protein family have emerged as IFN-induced proteins involved in both innate and adaptive immunity. In this report, we provide evidence that TRIM22 is a functional E3 ubiquitin ligase that is also ubiquitinated itself. We demonstrate that TRIM22 expression leads to a viral protection of HeLa cells against encephalomyocarditis virus infections. This effect is dependent upon its E3 ubiquitinating activity, since no antiviral effect was observed in cells expressing a TRIM22-deletion mutant defective in ubiquitinating activity. Consistent with this, TRIM22 interacts with the viral 3C protease (3C(PRO)) and mediates its ubiquitination. Altogether, our findings demonstrate that TRIM22 E3 ubiquitin ligase activity represents a new antiviral pathway induced by IFN against picornaviruses. PMID- 19218200 TI - Asymptomatically shed recombinant herpes simplex virus type 1 strains detected in saliva. AB - Herpes simplex virus type 1 (HSV-1) is a ubiquitous pathogen infecting most individuals worldwide. The majority of HSV-1-infected individuals have no clinical symptoms but shed HSV-1 asymptomatically in saliva. Recent phylogenetic analyses of HSV-1 have defined three genetic clades (A-C) and recombinants thereof. These data have all been based on clinical HSV-1 isolates and do not cover genetic variation of asymptomatically shed HSV-1. The primary goal of this study was to investigate such variation. A total of 648 consecutive saliva samples from five HSV-1-infected volunteers was collected. Asymptomatic shedding was detected on 7.6 % of the days from four subjects. The HSV-1 genome loads were quantified with real-time PCR and varied from 1x10(2) to 2.8x10(6) copies of virus DNA (ml saliva)(-1). Phylogenetic network analyses and bootscanning were performed on asymptomatically shed HSV-1. The analyses were based on DNA sequencing of the glycoprotein I gene, and also of the glycoprotein E gene for putative recombinants. For two individuals with clinical HSV-1 infection, the same HSV-1 strain was shed asymptomatically as induced clinical lesions, and sequence analyses revealed that these strains clustered distinctly to clades A and B, respectively. For one of the subjects with no clinical HSV-1 infection, a recombinant strain was identified. The other truly asymptomatic individual shed evolutionarily distinct HSV-1 strains on two occasions. The first strain was classified as a recombinant and the other strain clustered in clade A. High replication rates of different strains in the same person may facilitate the creation of recombinant clinical HSV-1 strains. PMID- 19218199 TI - Absence of spontaneous disease and comparative prion susceptibility of transgenic mice expressing mutant human prion proteins. AB - Approximately 15 % of human prion disease is associated with autosomal-dominant pathogenic mutations in the prion protein (PrP) gene. Previous attempts to model these diseases in mice have expressed human PrP mutations in murine PrP, but this may have different structural consequences. Here, we describe transgenic mice expressing human PrP with P102L or E200K mutations and methionine (M) at the polymorphic residue 129. Although no spontaneous disease developed in aged animals, these mice were readily susceptible to prion infection from patients with the homotypic pathogenic mutation. However, while variant Creutzfeldt-Jakob disease (CJD) prions transmitted infection efficiently to both lines of mice, markedly different susceptibilities to classical (sporadic and iatrogenic) CJD prions were observed. Prions from E200K and classical CJD M129 homozygous patients, transmitted disease with equivalent efficiencies and short incubation periods in human PrP 200K, 129M transgenic mice. However, mismatch at residue 129 between inoculum and host dramatically increased the incubation period. In human PrP 102L, 129M transgenic mice, short disease incubation periods were only observed with transmissions of prions from P102L patients, whereas classical CJD prions showed prolonged and variable incubation periods irrespective of the codon 129 genotype. Analysis of disease-related PrP (PrP(Sc)) showed marked alteration in the PrP(Sc) glycoform ratio propagated after transmission of classical CJD prions, consistent with the PrP point mutations directly influencing PrP(Sc) assembly. These data indicate that P102L or E200K mutations of human PrP have differing effects on prion propagation that depend upon prion strain type and can be significantly influenced by mismatch at the polymorphic residue 129. PMID- 19218201 TI - Cytomegaloviral protein kinase pUL97 interacts with the nuclear mRNA export factor pUL69 to modulate its intranuclear localization and activity. AB - Human cytomegalovirus encodes a number of phosphorylation-regulated proteins, including the autophosphorylating protein kinase pUL97 and the nuclear mRNA export factor pUL69. Recently, it was reported that the kinase inhibitor roscovitine induces an intranuclear aggregation of pUL69 in infected fibroblasts. Here, we demonstrate that pUL97-specific kinase inhibitors induce a similar pUL69 aggregation. Furthermore, a direct pUL69-pUL97 interaction was demonstrated by coimmunoprecipitation analyses. Deletion mapping identified the domains required for interaction in both proteins (1-140/478-532 in pUL69 and 231-336 in pUL97). Further analysis of the immunoprecipitates by in vitro kinase assays demonstrated the phosphorylation of pUL69 by pUL97. However, catalytically inactive mutants of pUL97 and interaction-negative fragments of pUL69 were phosphorylation-negative. Moreover, an analysis of the pUL69-mediated nuclear RNA export indicated a correlation of the export efficiency with the presence of active pUL97 kinase. These data suggest a specific pUL69-pUL97 interaction and pUL97-mediated phosphorylation which influences the regulatory activities of pUL69. PMID- 19218202 TI - Cytomegaloviral proteins that associate with the nuclear lamina: components of a postulated nuclear egress complex. AB - The nuclear egress of cytomegaloviral capsids traversing the nuclear envelope is dependent on a locally restricted destabilization of the rigid nuclear lamina. It has been suggested that the multi-component nuclear egress complex (NEC) that is formed is comprised of both viral and cellular proteins which act to recruit lamin-phosphorylating protein kinases. Recently, we reported that the lamina associated human cytomegalovirus-encoded proteins pUL50 and pUL53, conserved among herpesviruses, interact with each other and recruit protein kinase C (PKC) to the nuclear envelope in transfected cells. The multiple interactions of the transmembrane protein pUL50 with pUL53, PKC and cellular PKC-binding protein p32, appear crucial to the formation of the NEC. In this study, we mapped individual interaction sequence elements of pUL50 by coimmunoprecipitation analysis of deletion mutants and yeast two-hybrid studies. Amino acids 1-250 were shown to be responsible for interaction with pUL53, 100-280 for PKC and 100-358 for p32. Interestingly, p32 specifically interacted with multiple NEC components, including the kinases PKC and pUL97, thus possibly acting as an adaptor for protein recruitment to the lamin B receptor. Notably, p32 was the only protein that interacted with the lamin B receptor. Immunofluorescence studies visualized the colocalization of NEC components at the nuclear rim in coexpression studies. The data imply that a tight interaction between at least six viral and cellular proteins leads to the formation of a postulated multi-protein complex required for nuclear egress. PMID- 19218203 TI - Cleavage of Epstein-Barr virus glycoprotein B is required for full function in cell-cell fusion with both epithelial and B cells. AB - Glycoprotein B (gB) homologues within the herpesvirus family display high sequence conservation, and a number of gB homologues contain a cleavage motif R-X K/R-R recognized by the cellular protease furin. Epstein-Barr virus (EBV) gB contains this motif and cleaved gB is found in EBV virions. To determine the functional significance of this cleavage motif in EBV gB, a deletion mutant (gB Deltafurin) was created lacking the motif. This cleavage mutant was expressed well in cell culture but was not cleaved. Experiments examining gB Deltafurin in a cell-fusion assay revealed that fusion was reduced by 52 % in epithelial and 28 % in B cells when compared with wild-type EBV gB. This decrease in cell-cell fusion is similar to that observed with multiple alphaherpesvirus gB cleavage mutants and supports a conserved function for cleaved gB. PMID- 19218204 TI - Identification of a response element in a herpesvirus saimiri mRNA recognized by the ORF57 protein. AB - The herpesvirus saimiri (HVS) ORF57 protein binds viral RNA, enabling the efficient nuclear export of intronless viral mRNAs. However, it is not known how ORF57 recognizes these viral mRNAs. In this study, a systematic evolution of ligands by exponential enrichment (SELEX) approach was used to select RNA sequences that are preferentially bound by the ORF57 protein. Results identified a recurring motif, GAAGRG, within the majority of selected RNAs, which is also present in many late HVS mRNAs. RNA immunopreciptations demonstrated that disruption of this motif within a viral intronless RNA ablates ORF57 binding. These data suggest that the GAAGRG motif may be required within a HVS intronless mRNA for recognition by the ORF57 protein. PMID- 19218205 TI - In vivo importance of heparan sulfate-binding glycoproteins for murid herpesvirus 4 infection. AB - Many herpesviruses bind to heparan sulfate (HS). Murid herpesvirus-4 (MuHV-4) does so via its envelope glycoproteins gp70 and gH/gL. MuHV-4 gp150 further regulates an HS-independent interaction to make that HS-dependent too. Cell binding by MuHV-4 virions is consequently strongly HS-dependent. Gp70 and gH/gL show some in vitro redundancy: an antibody-mediated blockade of HS binding by one is well tolerated, whereas a blockade of both severely impairs infection. In order to understand the importance of HS binding for MuHV-4 in vivo, we generated mutants lacking both gL and gp70. As expected, gL(-)gp70(-) MuHV-4 showed very poor cell binding. It infected mice at high dose but not at low dose, indicating defective host entry. But once entry occurred, host colonization, which for MuHV 4 is relatively independent of the infection dose, was remarkably normal. The gL( )gp70(-) entry deficit was much greater than that of gL(-) or gp70(-) single knockouts. And gp150 disruption, which allows HS-independent cell binding, largely rescued the gL(-)gp70(-) cell binding and host entry deficits. Thus, it appeared that MuHV-4 HS binding is important in vivo, principally for efficient host entry. PMID- 19218206 TI - Investigation of orf virus structure and morphogenesis using recombinants expressing FLAG-tagged envelope structural proteins: evidence for wrapped virus particles and egress from infected cells. AB - Orf virus (ORFV) is the type species of the genus Parapoxvirus, but little is known about the structure or morphogenesis of the virus. In contrast, the structure and morphogenesis of vaccinia virus (VACV) has been extensively studied. VACV has two main infectious forms, mature virion (MV) and extracellular virion (EV). The MV is wrapped by two additional membranes derived from the trans Golgi to produce a wrapped virion (WV), the outermost of which is lost by cellular membrane fusion during viral egress to form the EV. Genome sequencing of ORFV has revealed that it has homologues of almost all of the VACV structural genes. Notable exceptions are A36R, K2L, A56R and B5R, which are associated with WV and EV envelopes. This study investigated the morphogenesis and structure of ORFV by fusing FLAG peptide to the structural proteins 10 kDa, F1L and ORF-110 to form recombinant viruses. 10 kDa and F1L are homologues of VACV A27L and H3L MV membrane proteins, whilst ORF-110 is homologous to VACV A34R, an EV membrane protein. Immunogold labelling of FLAG proteins on virus particles isolated from lysed cells showed that FLAG-F1L and FLAG-10 kDa were displayed on the surface of infectious particles, whereas ORF-110-FLAG could not be detected. Western blot analysis of solubilized recombinant ORF-110-FLAG particles revealed that ORF-110 FLAG was abundant and undergoes post-translational modification indicative of endoplasmic reticulum trafficking. Fluorescent microscopy confirmed the prediction that ORF-110-FLAG localized to the Golgi in virus-infected cells. Finally, immunogold labelling of EVs showed that ORF-110-FLAG became exposed on the surface of EV-like particles as a result of egress from the cell. PMID- 19218207 TI - Genomic characterization of the first insectivoran papillomavirus reveals an unusually long, second non-coding region and indicates a close relationship to Betapapillomavirus. AB - Knowledge about biological diversity is the prerequisite to reliably reconstruct the evolution of pathogens such as papillomaviruses (PV). However, complete genomes of non-human PV have only been cloned and sequenced from 8 out of 18 orders within the Placentalia, although the host-specific variety of PV is considered much larger. We isolated and sequenced the complete genome of the first insectivoran PV type from hair follicle cells of the European hedgehog (Erinaceus europaeus), designated EHPV. We conducted phylogenetic analyses (maximum-likelihood criterion and Bayesian inference) with the genomic information of a systematically representative set of 67 PV types including EHPV. As inferred from amino acid sequence data of the separate genes E1, E2 and L1 as well as of the gene combination E6-E7-E1-E2-L1, EHPV clustered within the beta gamma-pi-Xi-PV supertaxon and constituted the closest relative of genus Betapapillomavirus infecting primates. Beside the typical organization of the PV genome, EHPV exhibited a 1172 bp, non-coding region between the E2 and the L2 open reading frames. This trait has been previously described for the only distantly related Lambdapapillomavirus, but a common evolutionary origin of both non-coding regions is unlikely. Our results underscore the modular organization of the PV genome and the complex natural history of PV. PMID- 19218208 TI - Identification of species-specific and cross-reactive epitopes in human polyomavirus capsids using monoclonal antibodies. AB - The human antibody response to polyomavirus capsid proteins is not well characterized. Recombinant BK virus (BKV), JC virus (JCV) and simian virus 40 (SV40) virus-like particles (VLP) were produced in a baculovirus system, and mouse monoclonal antibodies (mAbs) to these proteins were generated using standard methods. Nine of 12 BKV mAbs showed neutralizing activity. The non neutralizing antibodies also bound BKV pseudocapsids in an ELISA binding assay. Most antibodies recognized conformational species-specific epitopes, but several exceptions were found: (i) BKV mAb BK-F11 cross-reacted with a linear buried epitope common to both JCV and SV40 pseudocapsids, (ii) two of six JCV antibodies (JC-6.7 and JC-7.9) and two of 13 SV40 antibodies (VP1-H2 and VP1-I2) recognized linear buried epitopes common to all three viruses and (iii) SV40 antibody VP1-E5 recognized a linear surface epitope on JCV pseudocapsids. PMID- 19218210 TI - Swine torque teno virus detection in pig commercial vaccines, enzymes for laboratory use and human drugs containing components of porcine origin. AB - Torque teno viruses (TTVs) are vertebrate infecting, single-stranded circular DNA viruses. Two genetically distinct TTV genogroups (TTV1 and TTV2) infect swine worldwide with high prevalence. Currently, swine TTVs are considered non pathogenic, although TTV2 has been linked to post-weaning multisystemic wasting syndrome, a porcine circovirus disease. On the other hand, pig materials are an important source of components used in porcine vaccine manufacturing, human drugs and commercial enzyme products. However, there is little information about the possible existence of extraneous viruses in products containing porcine-derived components. In the present study, 26 commercial swine vaccines, seven human drugs and three enzyme products from porcine origin were tested for the presence of TTV1 and TTV2 genomes by PCR. Four vaccines against Mycoplasma hyopneumoniae were positive for TTV2 by PCR. Three M. hyopneumoniae, one porcine parvovirus and one porcine reproductive and respiratory syndrome virus vaccines were PCR positive for TTV1. One human drug contained TTV1 DNA as well as a trypsin enzyme; a porcine-derived elastase product was positive for both TTV genogroups. These results show that swine TTVs are contaminants not only of swine vaccines but also of human drugs containing porcine components and enzymes for laboratory use. PMID- 19218209 TI - Assessment of recombinant beak and feather disease virus capsid protein as a vaccine for psittacine beak and feather disease. AB - Beak and feather disease virus (BFDV) is a significant pathogen of wild Australasian and African psittacine birds. We assessed the immunogenicity of recombinant BFDV capsid (recBFDVcap) to protect against the development of psittacine beak and feather disease (PBFD). Long-billed corellas (Cacatua tenuirostris) (n=13) received (by injection) 1 ml vaccine containing 10 microg recBFDVcap on day 0 and 0.4 ml vaccine containing 66.8 microg recBFDVcap on day 11. All vaccinated corellas and five non-vaccinated control corellas were given 0.4 ml BFDV suspension [titre=log(2) 12 haemagglutination units (HAU) 50 microl( 1)] intramuscularly and 0.1 ml orally 16 days after booster vaccination. Blood was collected during the vaccination period and blood and feathers were collected after BFDV administration. Testing of blood samples included BFDV DNA detection by PCR and quantitative PCR (qPCR) as well as antibody detection by haemagglutination inhibition (HI) and on feather samples, BFDV DNA and antigen was detected by haemagglutination (HA) and qPCR. Four of 97 blood samples collected from vaccinated birds after virus challenge tested positive by PCR, whereas 17 of 35 samples taken from non-vaccinated control corellas tested positive. Vaccinated birds did not develop feather lesions, had only transient PCR-detectable viraemia and had no evidence of persistent infection 270 days post challenge using PCR, histopathology and immunohistochemistry. Non-vaccinated control corellas developed transient feather lesions and had PCR, HI and HA test results consistent with PBFD. They were BFDV PCR-positive for up to 41 days post challenge and qPCR demonstrated reduced virus replication in vaccinated birds compared with non-vaccinated control birds. PMID- 19218211 TI - Caspase inhibitor P35 is required for the production of robust baculovirus virions in Trichoplusia ni TN-368 cells. AB - Apoptosis can protect lepidopteran insects against baculovirus infection by limiting viral replication. Baculoviruses counter this response by expressing anti-apoptotic proteins such as the caspase inhibitor P35, which is expressed by several baculoviruses including Autographa californica mutiple nucleopolyhedrovirus (AcMNPV). Mutants of AcMNPV that lack the p35 gene induce apoptosis in Spodoptera frugiperda cells, and replication of these mutants is severely curtailed in S. frugiperda cell lines and larvae. However, cells from another lepidopteran species, Trichoplusia ni, do not undergo apoptosis when infected with AcMNPV mutants lacking p35, and p35 mutant and wild-type viruses replicate to equivalent levels in the T. ni cell line TN-368 and have equivalent infectivity in T. ni larvae by either oral or intrahaemocoelic injection. This has led to the conclusion that p35 is not required for AcMNPV replication in T. ni. However, in this study it was found that stocks of p35 mutant virus produced in TN-368 cells had defects in virion stability and infectivity. TN-368 cells infected with p35 mutant AcMNPV exhibited caspase activity, despite a lack of apoptosis, and propagation of the mutant virus in the presence of a chemical caspase inhibitor restored the normal infection phenotype to the progeny virus. These results suggest that caspases can directly or indirectly damage baculovirus virions, and reveal a novel aspect of the role of apoptosis in antiviral defence. PMID- 19218212 TI - Diversity and evolution of the Cydia pomonella granulovirus. AB - Eight new field isolates of Cydia pomonella granulovirus (CpGV) originating in Iran and Georgia and one English CpGV isolate were analysed for restriction fragment length polymorphisms (RFLPs) and by partial genome amplification and sequencing. According to the observed RFLPs, most of the predominant genotypes of these isolates could be assigned to those present in previously found isolates originating from Mexico (CpGV-M), England (CpGV-E) and Russia (CpGV-R). We suggest that these isolates should be designated genome A, B and C types, respectively. A fourth genome type was identified in three isolates and is designated D type. The isolates with A, B and D type genomes contained four open reading frames (ORFs) (ORF63-ORF66) not present in C type genomes. The lack of these ORFs in other granuloviruses suggests that the C type genome is evolutionarily ancestral to the other genome types. The B and D type genomes contained an additional insertion of a non-protein coding region of 0.7 kb, which was at different genome locations. Analysis of the partial gene sequences of late expression factor 8 (lef-8), lef-9 and polyhedrin/granulin (polh/gran) genes revealed single nucleotide polymorphisms (SNPs) that corresponded to the RFLP types. Phylogenetic analyses based on these SNPs corroborated the proposed ancestry of the C type genome. C type viruses were also less virulent to neonate codling moth larvae than the other virus types. In conclusion, the known diversity of CpGV isolates can be described by four major genome types, which appear to exist in different isolates as genotype mixtures. PMID- 19218213 TI - Crystallographic structure of the alpha-helical triple coiled-coil domain of avian reovirus S1133 fibre. AB - Avian reovirus fibre, a homo-trimer of the sigmaC protein, is a minor component of the avian reovirus outer capsid. It is anchored via a short N-terminal sequence to the inner capsid lambdaC pentamer, and its protruding globular C terminal domain is responsible for primary host cell attachment. We have previously solved the structure of a receptor-binding fragment in which residues 160-191 form a triple beta-spiral and 196-326 a beta-barrel head domain. Here we have expressed, purified and crystallized a major sigmaC fragment comprising residues 117-326. Its structure, which was solved by molecular replacement using the previously determined receptor-binding domain structure and refined to 1.75 A (0.175 nm) resolution, reveals an alpha-helical triple coiled-coil connected to the previously solved structure by a zinc-ion-containing linker. The coiled-coil domain contains two chloride ion binding sites, as well as specific trimerization and registration sequences. The linker may act as a functionally important hinge. PMID- 19218214 TI - Genetic characterization of dengue virus type 1 isolated in Brunei in 2005-2006. AB - The full-length genomes of two DENV-1 viruses isolated during the 2005-2006 dengue incidents in Brunei were sequenced. Twenty five primer sets were designed to amplify contiguous overlapping fragments of approximately 500-600 base pairs spanning the entire sequence of the genome. The amplified PCR products were sent to a commercial laboratory for sequencing and the nucleotides and the deduced amino acids were determined. Sequence analysis of the envelope gene at the nucleotide and amino acid levels between the two isolates showed 92 and 96 % identity, respectively. Comparison of the envelope gene sequences with 68 other DENV-1 viruses of known genotypes placed the two isolates into two different genotypic groups. Isolate DS06/210505 belongs to genotype V together with some of the recent isolates from India (2003) and older isolates from Singapore (1990) and Burma (1976), while isolate DS212/110306 was clustered in genotype IV with the prototype Nauru strain (1974) and with some of the recent isolates from Indonesia (2004) and the Philippines (2002, 2001). In the full-length genome analysis at the nucleotide level, isolate DS06/210505 showed 94 % identity to the French Guyana strain (1989) in genotype V while isolate DS212/110306 had 96 % identity to the Nauru Island strain (1974) in genotype IV. This work constitutes the first complete genetic characterization of not only Brunei DENV-1 virus isolates, but also the first strain from Borneo Island. This study was the first to report the isolation of dengue virus in the country. PMID- 19218215 TI - Poly(A)-binding protein binds to the non-polyadenylated 3' untranslated region of dengue virus and modulates translation efficiency. AB - Poly(A)-binding protein (PABP) is a key player in mRNA circularization and translation initiation of polyadenylated mRNAs. It simultaneously binds the 3' poly(A) tail of an mRNA and eukaryotic initiation factor 4G (eIF4G), which forms part of the translation initiation complex assembling at the 5'end, thus circularizing the RNA molecule and enhancing translation initiation. Here, we report the binding of PABP to the non-polyadenylated 3'end of dengue virus (DENV) RNA. PABP binds the DENV 3' untranslated region (3'UTR) internally, upstream of the conserved 3'stem-loop near the two dumb-bell structures, and can be displaced by poly(A) RNA. The PABP-specific translation inhibitor PABP-interacting protein 2 (Paip2) interferes with the DENV 3'UTR-PABP interaction, and in vitro translation of DENV reporter RNAs in baby hamster kidney cell extracts is inhibited by Paip2 in a dose-dependent manner. Our findings show an expanded translation mechanism for PABP, binding to a viral RNA lacking a terminal poly(A) tail. PMID- 19218216 TI - Genetically engineered attenuated measles virus specifically infects and kills primary multiple myeloma cells. AB - The applicability of cytoreductive treatment of malignant diseases using recombinant viruses strongly depends on specific recognition of surface receptors to target exclusively neoplastic cells. A recently generated monoclonal antibody (mAb), Wue-1, specifically detects CD138(+) multiple myeloma (MM) cells. In this study, a haemagglutinin (H) protein that was receptor-blinded (i.e. did not bind to CD46 and CD150) was genetically re-engineered by fusing it to a single-chain antibody fragment (scFv) derived from the Wue-1 mAb open reading frame (scFv Wue), resulting in the recombinant retargeted measles virus (MV)-Wue. MV-Wue efficiently targeted and fully replicated in primary MM cells, reaching titres similar to those seen with non-retargeted viruses. In agreement with its altered receptor specificity, infection of target cells was no longer dependent on CD150 or CD46, but was restricted to cells that had been labelled with Wue-1 mAb. Importantly, infection with MV-Wue rapidly induced apoptosis in CD138(+) malignant plasma cell targets. MV-Wue is the first fully retargeted MV using the restricted interaction between Wue-1 mAb and primary MM cells specifically to infect, replicate in and deplete malignant plasma cells. PMID- 19218217 TI - Porcine endogenous retroviruses PERV A and A/C recombinant are insensitive to a range of divergent mammalian TRIM5alpha proteins including human TRIM5alpha. AB - The potential risk of cross-species transmission of porcine endogenous retroviruses (PERV) to humans has slowed the development of xenotransplantation, using pigs as organ donors. Here, we show that PERVs are insensitive to restriction by divergent TRIM5alpha molecules despite the fact that they strongly restrict a variety of divergent lentiviruses. We also show that the human PERV A/C recombinant clone 14/220 reverse transcribes with increased efficiency in human cells, leading to significantly higher infectivity. We conclude that xenotransplantation studies should consider the danger of highly infectious TRIM5alpha-insensitive human-tropic PERV recombinants. PMID- 19218218 TI - Complexity in human immunodeficiency virus type 1 (HIV-1) co-receptor usage: roles of CCR3 and CCR5 in HIV-1 infection of monocyte-derived macrophages and brain microglia. AB - CCR3 has been implicated as a co-receptor for human immunodeficiency virus type 1 (HIV-1), particularly in brain microglia cells. We sought to clarify the comparative roles of CCR3 and CCR5 in the central nervous system (CNS) HIV-1 infection and the potential utility of CCR3 as a target for manipulation via gene transfer. To target CCR3, we developed a single-chain antibody (SFv) and an interfering RNA (RNAi), R3-526. Coding sequences for both were cloned into Tag deleted SV40-dervied vectors, as these vectors transduce brain microglia and monocyte-derived macrophages (MDM) highly efficiently. These anti-CCR3 transgenes were compared to SFv-CCR5, an SFv against CCR5, and RNAi-R5, an RNAi that targets CCR5, for the ability to protect primary human brain microglia and MDM from infection with peripheral and neurotropic strains of HIV-1. Downregulation of CCR3 and CCR5 by these transgenes was independent from one another. Confocal microscopy showed that CCR3 and CCR5 co-localized at the plasma membrane with each other and with CD4. Targeting either CCR5 or CCR3 largely protected both microglia and MDM from infection by many strains of HIV-1. That is, some HIV-1 strains, isolated from either the CNS or periphery, required both CCR3 and CCR5 for optimal productive infection of microglia and MDM. Some HIV-1 strains were relatively purely CCR5-tropic. None was purely CCR3-tropic. Thus, some CNS-tropic strains of HIV-1 utilize CCR5 as a co-receptor but do not need CCR3, while for other isolates both CCR3 and CCR5 may be required. PMID- 19218219 TI - Role of the zinc-finger and basic motifs of chrysanthemum virus B p12 protein in nucleic acid binding, protein localization and induction of a hypersensitive response upon expression from a viral vector. AB - The genomes of carlaviruses encode cysteine-rich proteins (CRPs) of unknown function. The 12 kDa CRP of chrysanthemum virus B (CVB), p12, has been shown previously to induce a hypersensitive response (HR) when expressed from potato virus X (PVX). This study demonstrated that a p12-induced HR was preceded by induction of a number of genes related to pathogenesis, stress and systemic acquired resistance. p12 localized predominantly to the nucleus. Interestingly, it was found that p12 bound both RNA and DNA in vitro, but notably exhibited a preference for DNA in the presence of Zn(2+) ions. Mutational analysis of the p12 conserved sequence motifs demonstrated that the basic motif is required for p12 translocation to the nucleus, thus representing part of the protein nuclear localization signal, whereas the predicted zinc finger motif is needed for both Zn(2+)-dependent DNA binding and eliciting an HR in PVX-infected leaves. Collectively, these results link, for the first time, nuclear localization of the protein encoded by a cytoplasmically replicating virus and its DNA-binding capacity with HR induction. Furthermore, these data suggest that p12 may mediate induction of the host genes by binding to the plant genomic DNA, and emphasize that CVB p12 is functionally distinct from other known nuclear-localized proteins encoded by the plant positive-stranded RNA viruses. PMID- 19218220 TI - Rapid host adaptation by extensive recombination. AB - Experimental investigations into virus recombination can provide valuable insights into the biochemical mechanisms and the evolutionary value of this fundamental biological process. Here, we describe an experimental scheme for studying recombination that should be applicable to any recombinogenic viruses amenable to the production of synthetic infectious genomes. Our approach is based on differences in fitness that generally exist between synthetic chimaeric genomes and the wild-type viruses from which they are constructed. In mixed infections of defective reciprocal chimaeras, selection strongly favours recombinant progeny genomes that recover a portion of wild-type fitness. Characterizing these evolved progeny viruses can highlight both important genetic fitness determinants and the contribution that recombination makes to the evolution of their natural relatives. Moreover, these experiments supply precise information about the frequency and distribution of recombination breakpoints, which can shed light on the mechanistic processes underlying recombination. We demonstrate the value of this approach using the small single-stranded DNA geminivirus, maize streak virus (MSV). Our results show that adaptive recombination in this virus is extremely efficient and can yield complex progeny genomes comprising up to 18 recombination breakpoints. The patterns of recombination that we observe strongly imply that the mechanistic processes underlying rolling circle replication are the prime determinants of recombination breakpoint distributions found in MSV genomes sampled from nature. PMID- 19218221 TI - Genome activation by raspberry bushy dwarf virus coat protein. AB - Two sets of infectious cDNA clones of raspberry bushy dwarf virus (RBDV) have been constructed, enabling either the synthesis of infectious RNA transcripts or the delivery of infectious binary plasmid DNA by infiltration of Agrobacterium tumefaciens. In whole plants and in protoplasts, inoculation of RBDV RNA1 and RNA2 transcripts led to a low level of infection, which was greatly increased by the addition of RNA3, a subgenomic RNA coding for the RBDV coat protein (CP). Agroinfiltration of RNA1 and RNA2 constructs did not produce a detectable infection but, again, inclusion of a construct encoding the CP led to high levels of infection. Thus, RBDV replication is greatly stimulated by the presence of the CP, a mechanism that also operates with ilarviruses and alfalfa mosaic virus, where it is referred to as genome activation. Mutation to remove amino acids from the N terminus of the CP showed that the first 15 RBDV CP residues are not required for genome activation. Other experiments, in which overlapping regions at the CP N terminus were fused to the monomeric red fluorescent protein, showed that sequences downstream of the first 48 aa are not absolutely required for genome activation. PMID- 19218222 TI - The beet virus Q coat protein readthrough domain is longer than previously reported, with two transmembrane domains. AB - Ten beet virus Q (BVQ) strains from six different countries were sequenced to characterize the readthrough (RT) domain of the coat protein (CP). The GenBank/EMBL/DDBJ accession numbers for the sequences reported in this paper are FM244643-FM244652. With three nucleotide additions of 5, 285 and 1 nt, the common RT of 76 kDa was found to be longer than the single reference available to date (35 kDa). It is hypothesized that multiple inoculation cycles on Chenopodium quinoa were responsible for these three deletions in the C-terminal part of the BVQ RNA-2 previously described. Two putative transmembrane domains, TM1 and TM2, were predicted in the consensus amino acid sequence of the ten BVQ strains, and the putative BVQ TM2 was aligned with that of potato mop-top virus. PMID- 19218223 TI - A single U/C nucleotide substitution changing alanine to valine in the beet necrotic yellow vein virus P25 protein promotes increased virus accumulation in roots of mechanically inoculated, partially resistant sugar beet seedlings. AB - Beet necrotic yellow vein virus (BNYVV) A type isolates E12 and S8, originating from areas where resistance-breaking had or had not been observed, respectively, served as starting material for studying the influence of sequence variations in BNYVV RNA 3 on virus accumulation in partially resistant sugar beet varieties. Sub-isolates containing only RNAs 1 and 2 were obtained by serial local lesion passages; biologically active cDNA clones were prepared for RNAs 3 which differed in their coding sequences for P25 aa 67, 68 and 129. Sugar beet seedlings were mechanically inoculated with RNA 1+2/RNA 3 pseudorecombinants. The origin of RNAs 1+2 had little influence on virus accumulation in rootlets. E12 RNA 3 coding for V(67)C(68)Y(129) P25, however, enabled a much higher virus accumulation than S8 RNA 3 coding for A(67)H(68)H(129) P25. Mutants revealed that this was due only to the V(67) 'GUU' codon as opposed to the A(67) 'GCU' codon. PMID- 19218224 TI - Three serial passages of bovine spongiform encephalopathy in sheep do not significantly affect discriminatory test results. AB - During the 1980s, bovine spongiform encephalopathy (BSE)-contaminated meat and bonemeal were probably fed to sheep, raising concerns that BSE may have been transmitted to sheep in the UK. The human disease, variant Creutzfeldt-Jakob disease, arose during the BSE epidemic, and oral exposure of humans to BSE infected tissues has been implicated in its aetiology. The concern is that sheep BSE could provide another source of BSE exposure to humans via sheep products. Two immunological techniques, Western immunoblotting (WB) and immunohistochemistry (IHC), have been developed to distinguish scrapie from cases of experimental sheep BSE by the characteristics of their respective abnormal, disease-associated prion proteins (PrP(d)). This study compares the WB and IHC characteristics of PrP(d) from brains of primary, secondary and tertiary experimental ovine BSE cases with those of cattle BSE and natural sheep scrapie. Discrimination between experimental sheep BSE and scrapie remained possible by both methods, regardless of the route of challenge. PMID- 19218225 TI - Identification of seven haplotypes of the caprine PrP gene at codons 127, 142, 154, 211, 222 and 240 in French Alpine and Saanen breeds and their association with classical scrapie. AB - In sheep, susceptibility to scrapie is mainly influenced by polymorphisms of the PrP gene. In goats, there are to date few data related to scrapie susceptibility association with PrP gene polymorphisms. In this study, we first investigated PrP gene polymorphisms of the French Alpine and Saanen breeds. Based on PrP gene open reading frame sequencing of artificial insemination bucks (n=404), six encoding mutations were identified at codons 127, 142, 154, 211, 222 and 240. However, only seven haplotypes could be detected: four (GIH(154)RQS, GIRQ(211)QS, GIRRK(222)S and GIRRQP(240)) derived from the wild-type allele (G(127)I(142)R(154)R(211)Q(222)S(240)) by a single-codon mutation, and two (S(127)IRRQP(240) and GM(142)RRQP(240)) by a double-codon mutation. A case control study was then implemented in a highly affected Alpine and Saanen breed herd (90 cases/164 controls). Mutations at codon 142 (I/M), 154 (R/H), 211 (R/Q) and 222 (Q/K) were found to induce a significant degree of protection towards natural scrapie infection. Compared with the baseline homozygote wild-type genotype I(142)R(154)R(211)Q(222)/IRRQ goats, the odds of scrapie cases in IRQ(211)Q/IRRQ and IRRK(222)/IRRQ heterozygous animals were significantly lower [odds ratio (OR)=0.133, P<0.0001; and OR=0.048, P<0.0001, respectively]. The heterozygote M(142)RRQ/IRRQ genotype was only protective (OR=0.243, P=0.0186) in goats also PP(240) homozygous at codon 240. However, mutated allele frequencies in French Alpine and Saanen breeds were low (0.5-18.5 %), which prevent us from assessing the influence of all the possible genotypes in natural exposure conditions. PMID- 19218226 TI - Unswitched immunoglobulin M response prolongs mouse survival in prion disease. AB - Several studies have failed to demonstrate the presence of immune responses to infectious prions during the course of prion disease, reflecting the identical primary structure of normal and disease-associated isoforms and the widespread expression of the normal cellular form of prion protein, PrP(C), leading to B- and/or T-cell tolerance of disease-associated isoforms and also possibly because antigen-presenting cells are unable to process the highly aggregated, detergent insoluble, protease-resistant form, PrP(Sc). Under certain circumstances, PrP(Sc) can be revealed to the immune system in immunogenic form, and it has been shown previously that anti-PrP antibodies can be induced to prions immunoadsorbed to Dynabeads using specific anti-PrP monoclonal antibodies, even in PrP-sufficient mice. This study demonstrated in a murine scrapie model that PrP-Dynabeads effectively stimulated the immune system to produce anti-PrP IgM antibodies over prolonged periods after repeated immunization. It was also shown that these immune responses prolonged incubation times in murine scrapie. PMID- 19218227 TI - A role for preoptic glutamate in the regulation of male reproductive behavior. AB - Although much progress has been made toward understanding the role of the medial preoptic area (MPOA) in the regulation of male reproductive behaviors, the precise mechanisms responsible for its activation during mating are largely unclear. Several studies implicate glutamate in this response. However, not until recently was there direct evidence supporting this hypothesis. Results obtained using in vivo microdialysis showed that levels of glutamate increased in the MPOA during mating, particularly with ejaculation. Levels then decreased rapidly following ejaculation, during a period of sexual quiescence. The magnitude of this decrease correlated with time spent in quiescence. Additionally, central administration of glutamate uptake inhibitors increased levels of glutamate and facilitated behavior. Glutamate activation of N-methyl-D-aspartate (NMDA) receptors in the MPOA is at least partly responsible for behavioral effects evoked by increase glutamate. This is evidenced by histological analysis of the MPOA, which shows that nearly all cells containing mating-induced Fos also contained NMDA receptors. Mating also increased phosphorylation of NMDA receptors, indicating receptor activation. Finally, bilateral microinjections of NMDA receptor antagonists inhibited copulation. This neurochemical, anatomical, and behavioral evidence points to a key role of preoptic glutamate in the regulation of sexual behavior in males. The implications of these findings are discussed. PMID- 19218229 TI - Whole-body responses: neural control and implications for rehabilitation and fall prevention. AB - Humans are one of the unique species that utilize bipedal gait to ambulate in our environment. Despite this fact, coordination of the arms with the legs and the rest of body is essential for many daily activities. As such, whole-body responses have emerged as the preferred strategy following perturbations to balance during both standing and walking. Complex neural circuitry may allow for this coordination through the use of propriospinal pathways linking lumbar and cervical pattern generators in the spinal cord, with supraspinal centers altering this control depending on the context of the situation. Based on these findings, we argue that whole-body reactions may be exploited for rehabilitation purposes. Preliminary results have indicated training programs designed to elicit whole body responses are effective in reducing falls and improving functional mobility in older adults with and without neurological impairment. PMID- 19218228 TI - From neurotransmitters to neurotrophic factors to neurogenesis. AB - New neurons continue to be produced in adult mammals, including humans, predominantly in the anterior subventricular zone of the lateral ventricle and the subgranular zone of the dentate gyrus. This update focuses on the emerging concept that adult CNS neurogenesis can be regulated by targeting neurotransmitter receptors, which, in turn, drive expression of crucial neurotrophic and growth factors. Such an approach might enable the development of pharmacological treatments that harness the endogenous potential of the CNS to replace lost cells in neurological disorders such as stroke and Alzheimer's and Huntington's diseases. This review samples in vivo studies in adult mammals from 2006 to mid-2008. It also provides some considerations for navigating toward translation to human disorders. Among them are the formidable problems of scaling up production of new neurons within the two "niches" of the brain and delivering sufficient numbers to distant degenerating regions for cell replacement. However, an expedition can only succeed if started. PMID- 19218230 TI - Neurotrophic support and oxidative stress: converging effects in the normal and diseased nervous system. AB - Oxidative stress and loss of neurotrophic support play major roles in the development of various diseases of the central and peripheral nervous systems. In disorders of the central nervous system such as Alzheimer's, Parkinson's, and Huntington's diseases, oxidative stress appears inextricably linked to the loss of neurotrophic support. A similar situation is seen in the peripheral nervous system in diseases of olfaction, hearing, and vision. Neurotrophic factors act to up-regulate antioxidant enzymes and promote the expression of antioxidant proteins. On the other hand, oxidative stress can cause down-regulation of neurotrophic factors. We propose that normal functioning of the nervous systems involves a positive feedback loop between antioxidant processes and neurotrophic support. Breakdown of this feedback loop in disease states leads to increased oxidative stress and reduced neurotrophic support. PMID- 19218231 TI - Ocular dominance columns: enigmas and challenges. AB - In some mammalian species, geniculocortical afferents serving each eye are segregated in layer 4C of striate cortex into stripes called ocular dominance columns. Having described the complete pattern of ocular dominance columns in the human brain, the authors enumerate here the principal enigmas that confront future investigators. Probably the overarching challenge is to explain the function, if any, of ocular dominance columns and why they are present in some species and not others. A satisfactory solution must account for the enormous natural variation, even within the same species, among individuals in column expression, pattern, periodicity, and alignment with other components of the functional architecture. Another major priority is to explain the development of ocular dominance columns. It has been established clearly that they form without visual experience, but the innate signals that guide their segregation and maturation are unknown. Experiments addressing the role of spontaneous retinal activity have yielded contradictory data. These studies must be reconciled, to pave the way for new insights into how columnar structure is generated in the cerebral cortex. PMID- 19218232 TI - Molecular motors and synaptic assembly. AB - Proper synaptic function requires the seamless integration of the transport, assembly, and regulation of synaptic components and structures. Inasmuch as the synapse is often distant from the neuronal cell body, newly synthesized synaptic proteins, the precursors of synaptic vesicles, active zone compartments, channels and receptors, and mitochondria, must be transported along lengthy neuronal processes to participate in synaptogenesis. Neuronal transport is mediated by motor proteins that associate with their cargoes via adaptors (or receptors) and that travel along the cytoskeleton network within the neuronal processes. Thus, the identity of membranous protein cargoes and the specificity of motor-cargo interactions are critical for correctly targeting cargoes and properly assembling synapses in developing neurons and in remodeling synapses of mature neurons in response to neuronal activity. In this article, the authors review recent progress in characterizing microtubule- and actin-based motor proteins that are involved in delivering synaptic components and discuss potential mechanisms underlying the formation of motor-receptor-cargo complexes that contribute to synaptogenesis and activity-induced synaptic plasticity. PMID- 19218233 TI - Transducing neuronal activity into dendritic spine morphology: new roles for p38 MAP kinase and N-cadherin. AB - Synaptic plasticity depends on the generation, modification and disconnection of synapses. An excitatory synapse is connected to a specialized dendritic compartment called a spine, which undergoes activity-induced remodeling. Here, we discuss a signaling pathway that transduces neuronal activity into the remodeling of spine through p38 mitogen-activated protein kinase (MAPK) and N-cadherin. Dendritic spines change their morphology and density in response to neuronal activity. In the early phase, posttranslational modifications of synaptic molecules regulate spine morphology, whereas activity-induced gene products reduce spine density in the late phase. One of the targets of these mechanisms is N-cadherin. An activity-induced protocadherin, arcadlin, stimulates thousand and one 2beta (TAO2beta) kinase, which in turn activates p38 MAPK through MAPK kinase 3 (MEK3), resulting in the endocytosis of N-cadherin and the decrease in spine number. This pathway also underlies the mechanism of the spine decrease in neuronal disorders, such as Alzheimer's disease and epilepsy. Development of new p38 MAPK inhibitors brings a ray of hope with respect to the development of more effective therapies for these patients. PMID- 19218234 TI - Neurotrophic modulation of motor neuron development. AB - Neurotrophic factors (NTFs) are a pleiotropic group of secreted growth factors that regulate multiple aspects of neuronal development, including the regressive event of cell death. Skeletal muscleinnervating lower motoneurons (MNs) of the brain stem and spinal cord comprise one population of central neurons in which programmed cell death (PCD) during embryogenesis has been actively investigated, as much for reasons of technical facility as clinical relevance. The precise identity of NTF-dependent MNs has remained unclear, with most studies simply reporting losses or gains across the entire spinal cord or individual brain-stem nuclei. However, MNs are grouped into highly heterogenous populations based on transcriptional identity, target innervation, and physiological function. Therefore, recent work has focused on the effects of NTF overexpression or deletion on the survival of these MN subpopulations. Together with the recent progress attained in the generation of conditional mutant mice, in which the function of an NTF or its receptor can be eliminated specifically in MNs, these recent studies have begun to define the differential trophic requirements for MN subpopulations during PCD. The intent of this review is to summarize these recent findings and to discuss their significance with respect to neurotrophic theory. PMID- 19218237 TI - Characterization of N-terminal structure of TLR2-activating lipoprotein in Staphylococcus aureus. AB - Staphylococcus aureus is known to activate mammalian immune cells through Toll like receptor 2 (TLR2). We recently demonstrated that a lipoprotein fraction obtained from S. aureus by Triton X-114 phase partitioning is a potent activator of TLR2. In this study, we separated TLR2-activating lipoproteins expressed in S. aureus and characterized an N-terminal structure. The lipoprotein fraction of S. aureus was prepared by glass bead disruption followed by Triton X-114 phase partitioning. The TLR2-activating molecules were mainly detected in the mass range of 30-35 kDa. Seven lipoproteins were identified by the mass spectra of their tryptic digests. Among them, three lipoproteins were separated by preparative SDS-PAGE and proved to activate TLR2. After digestion with trypsin in the presence of sodium deoxycholate, the N terminus of the lipopeptide was isolated from lipoprotein SAOUHSC_02699 by normal phase high pressure liquid chromatography and characterized as an S-(diacyloxypropyl)cystein-containing peptide using tandem mass spectra. The synthetic lipopeptide counterpart also stimulated the cells via TLR2. These results showed that the diacylated lipoprotein from S. aureus acts as a TLR2 ligand in mammalian cells. PMID- 19218236 TI - Inhibition of SUV39H1 methyltransferase activity by DBC1. AB - SUV39H1 is a histone H3K9-specific methyltransferase important for heterochromatin formation, regulation of gene expression, and induction of senescence in premalignant cells. SUV39H1 forms a complex with SirT1, and its activity is stimulated by SirT1 binding. Here we present evidence that the product of the DBC1 (deleted in breast cancer 1) gene disrupts the SUV39H1-SirT1 complex. Furthermore, DBC1 binds to the SUV39H1 catalytic domain and inhibits its ability to methylate histone H3 in vitro and in vivo. Knockdown of endogenous DBC1 increased the level of cellular H3K9 methylation. As expected, DBC1 also binds to SirT1 and inhibits the deacetylase activity of SirT1. These results identify DBC1 as a novel cellular inhibitor of SUV39H1 activity. DBC1 may be an important regulator of heterochromatin formation and genomic stability by disrupting the SUV39H1-SirT1 complex and inactivating both enzymes. PMID- 19218238 TI - Intranuclear degradation of polyglutamine aggregates by the ubiquitin-proteasome system. AB - Huntington disease and its related autosomal-dominant polyglutamine (pQ) neurodegenerative diseases are characterized by intraneuronal accumulation of protein aggregates. Studies on protein aggregates have revealed the importance of the ubiquitin-proteasome system as the front line of protein quality control (PQC) machinery against aberrant proteins. Recently, we have shown that the autophagy-lysosomal system is also involved in cytoplasmic aggregate degradation, but the nucleus lacked this activity. Consequently, the nucleus relies entirely on the ubiquitin-proteasome system for PQC. According to previous studies, nuclear aggregates possess a higher cellular toxicity than do their cytoplasmic counterparts, however degradation kinetics of nuclear aggregates have been poorly understood. Here we show that nuclear ubiquitin ligases San1p and UHRF-2 each enhance nuclear pQ aggregate degradation and rescued pQ-induced cytotoxicity in cultured cells and primary neurons. Moreover, UHRF-2 is associated with nuclear inclusion bodies in vitro and in vivo. Our data suggest that UHRF-2 is an essential molecule for nuclear pQ degradation as a component of nuclear PQC machinery in mammalian cells. PMID- 19218239 TI - The Gcn5 bromodomain of the SAGA complex facilitates cooperative and cross-tail acetylation of nucleosomes. AB - Bromodomains are acetyl lysine binding modules found in many complexes that regulate gene transcription. In budding yeast, the coactivator complex SAGA (Spt Ada-Gcn5-acetyl-transferase) predominantly facilitates transcription of stress activated genes and requires the bromodomain of the Gcn5 subunit for full activation of a number of these genes. This bromodomain has previously been shown to promote retention of the complex to H3 and H4 acetylated nucleosomes. Because the SAGA complex mediates histone H3 acetylation, we sought to determine to what extent the Gcn5 bromodomain directly modulates histone acetylation activity. Kinetic analysis of SAGA-mediated acetylation of nucleosomal substrates reveals that this bromodomain: 1) is required for the cooperative acetylation of nucleosomes, 2) enhances acetylation of an H3 histone tail when the other H3 tail within a nucleosome is already acetylated, and 3) augments the acetylation turnover of nucleosomes previously acetylated at lysine 16 of the histone H4 tails. These results indicate that the Gcn5 bromodomain promotes the establishment of nucleosome acetylation through multiple mechanisms and more generally show how chromatin recognition domains can modulate the enzymatic activity of chromatin modifying complexes. PMID- 19218240 TI - BLNK binds active H-Ras to promote B cell receptor-mediated capping and ERK activation. AB - Cross-linked B cell receptor (BCR) aggregates on the cell surface, then assembles into the "cap" where Ras is co-localized, and transduces various intracellular signals including Ras-ERK activation. BCR signals induce proliferation, differentiation, or apoptosis of B cells depending on their maturational stage. The adaptor protein BLNK binds various signaling proteins and Igalpha, a signaling subunit of the BCR complex, and plays an important role in the BCR signal transduction. BLNK was shown to be required for activation of ERK, but not of Ras, after BCR cross-linking, raising a question how BLNK facilitates ERK activation. Here we demonstrate that BLNK binds the active form of H-Ras, and their binding is facilitated by BCR cross-linking. We have identified a 10-amino acid Ras-binding domain within BLNK that is necessary for restoration of BCR mediated ERK activation in BLNK-deficient B cells and for anti-apoptotic signaling. The Ras-binding domain fused with a CD8alpha-Igalpha chimeric receptor could induce prolonged ERK phosphorylation, transcriptional activation of Elk1, as well as the capping of the receptor in BLNK-deficient B cells. These results indicate that BLNK recruits active H-Ras to the BCR complex, which is essential for sustained surface expression of BCR in the form of the cap and for the signal leading to functional ERK activation. PMID- 19218241 TI - Peroxisome proliferator-activated receptor {gamma} stimulation of adipocyte ApoE gene transcription mediated by the liver receptor X pathway. AB - Peroxisome proliferator-activated receptor (PPARgamma) agonists increase insulin sensitivity in humans and are useful for treating human diabetes. Treatment with these agonists leads to increased apoE expression and triglyceride accumulation in adipocytes. The importance of apoE for adipocyte triglyceride accumulation is demonstrated by observations that triglyceride accumulation is impaired in apoE knockout adipocytes treated with PPARgamma agonists. The current studies investigate the molecular mechanism for PPARgamma stimulation of the adipocyte apoE gene and demonstrate that the liver receptor X (LXR) response element within an apoE gene downstream enhancer is required for the apoE response to PPARgamma agonists. The response of the apoE gene to treatment with PPARgamma agonists was delayed beyond 12 h suggesting the involvement of an intermediary pathway. The combined addition of PPARgamma and LXR agonists did not increase apoE response beyond that observed with addition of either alone. Deletion or mutation of the LXR response element completely eliminated the adipocyte apoE gene response to a PPARgamma agonist. Chromatin immunoprecipitation analyses performed using isolated adipocytes, or adipose tissue from mice treated with PPARgamma agonists, showed increased LXR binding to the apoE gene after PPARgamma agonist treatment. Knockdown of LXR expression completely eliminated the increase in apoE message, protein, and triglyceride in response to PPARgamma stimulation. The LXR response element has been previously shown to mediate sterol responsiveness of the apoE gene, and apoE expression plays an important role in adipocyte triglyceride balance. The current observations suggest that the PPARgamma-LXR-apoE regulatory cascade could be an important molecular link for cross-talk between adipocyte triglyceride and cholesterol homeostasis. PMID- 19218242 TI - Subunit interactions and composition of the fructose 6-phosphate catalytic site and the fructose 2,6-bisphosphate allosteric site of mammalian phosphofructokinase. AB - Mammalian phosphofructokinase originated by duplication, fusion, and divergence of a primitive prokaryotic gene, with the duplicated fructose 6-phosphate catalytic site in the C-terminal half becoming an allosteric site for the activator fructose 2,6-bisphosphate. It has been suggested that both sites are shared across the interface between subunits aligned in an antiparallel orientation, the N-terminal half of one subunit facing the C-terminal half of the other. The composition of these binding sites and the way in which subunits interact to form the dimer within the tetrameric enzyme have been reexamined by systematic point mutations to alanine of key amino acid residues of human muscle phosphofructokinase. We found that residues His-199, His-298, Arg-201, and Arg 292 contribute to the catalytic site and not to the allosteric site, because their mutation decreased the affinity for fructose 6-phosphate without affecting the activation by fructose 2,6-bisphosphate or its binding affinity. In contrast, residues Arg-566, Arg-655, and His-661 were critical components of the fructose bisphosphate allosteric site, because their mutation strongly reduced the action and affinity of the activator, with no alteration of substrate binding to the active site. Our results suggest that mammalian phosphofructokinase subunits associate with the N-terminal halves facing each other to form the two catalytic sites/dimer and the C-terminal halves forming the allosteric sites. Additionally, mutation of certain residues eliminated activation by fructose 1,6-bisphosphate, but not its binding, with little effect on activation by fructose 2,6 bisphosphate, indicating a divergence in the signal transduction route despite their binding to the same site. PMID- 19218243 TI - The cardiac IKs potassium channel macromolecular complex includes the phosphodiesterase PDE4D3. AB - The cardiac I(Ks) potassium channel is a macromolecular complex consisting of alpha-(KCNQ1) and beta-subunits (KCNE1) and the A kinase-anchoring protein (AKAP) Yotiao (AKAP-9), which recruits protein kinase A) and protein phosphatase 1 to the channel. Here, we have tested the hypothesis that specific cAMP phosphodiesterase (PDE) isoforms of the PDE4D family that are expressed in the heart are also part of the I(Ks) signaling complex and contribute to its regulation by cAMP. PDE4D isoforms co-immunoprecipitated with I(Ks) channels in hearts of mice expressing the I(Ks) channel. In myocytes isolated from these mice, I(Ks) was increased by pharmacological PDE inhibition. PDE4D3, but not PDE4D5, co-immunoprecipitated with the I(Ks) channel only in Chinese hamster ovary cells co-expressing AKAP-9, and PDE4D3, but not PDE4D5, co immunoprecipitated with AKAP-9. Functional experiments in Chinese hamster ovary cells expressing AKAP-9 and either PDE4D3 or PDE4D5 isoforms revealed modulation of the I(Ks) response to cAMP by PDE4D3 but not PDE4D5. We conclude that PDE4D3, like protein kinase A and protein phosphatase 1, is recruited to the I(Ks) channel via AKAP-9 and contributes to its critical regulation by cAMP. PMID- 19218245 TI - Transforming growth factor-{beta}-inducible phosphorylation of Smad3. AB - Smad proteins transduce the transforming growth factor-beta (TGF-beta) signal at the cell surface into gene regulation in the nucleus. Upon TGF-beta treatment, the highly homologous Smad2 and Smad3 are phosphorylated by the TGF-beta receptor at the SSXS motif in the C-terminal tail. Here we show that in addition to the C tail, three (S/T)-P sites in the Smad3 linker region, Ser(208), Ser(204), and Thr(179) are phosphorylated in response to TGF-beta. The linker phosphorylation peaks at 1 h after TGF-beta treatment, behind the peak of the C-tail phosphorylation. We provide evidence suggesting that the C-tail phosphorylation by the TGF-beta receptor is necessary for the TGF-beta-induced linker phosphorylation. Although the TGF-beta receptor is necessary for the linker phosphorylation, the receptor itself does not phosphorylate these sites. We further show that ERK is not responsible for TGF-beta-dependent phosphorylation of these three sites. We show that GSK3 accounts for TGF-beta-inducible Ser(204) phosphorylation. Flavopiridol, a pan-CDK inhibitor, abolishes TGF-beta-induced phosphorylation of Thr(179) and Ser(208), suggesting that the CDK family is responsible for phosphorylation of Thr(179) and Ser(208) in response to TGF-beta. Mutation of the linker phosphorylation sites to nonphosphorylatable residues increases the ability of Smad3 to activate a TGF-beta/Smad-target gene as well as the growth-inhibitory function of Smad3. Thus, these observations suggest that TGF-beta-induced phosphorylation of Smad3 linker sites inhibits its antiproliferative activity. PMID- 19218244 TI - Control of alternative splicing by signal-dependent degradation of splicing regulatory proteins. AB - Alternative pre-mRNA splicing is a major gene expression regulatory mechanism in metazoan organisms. Proteins that bind pre-mRNA elements and control assembly of splicing complexes regulate utilization of pre-mRNA alternative splice sites. To understand how signaling pathways impact this mechanism, an RNA interference screen in Drosophila S2 cells was used to identify proteins that regulate TAF1 (TBP-associated factor 1) alternative splicing in response to activation of the ATR (ATM-RAD3-related) signaling pathway by the chemotherapeutic drug camptothecin (CPT). The screen identified 15 proteins that, when knocked down, caused the same change in TAF1 alternative splicing as CPT treatment. However, combined RNA interference and CPT treatment experiments indicated that only a subset of the identified proteins are targets of the CPT-induced signal, suggesting that multiple independent pathways regulate TAF1 alternative splicing. To understand how signals modulate the function of splicing factors, we characterized one of the CPT targets, Tra2 (Transformer-2). CPT was found to down regulate Tra2 protein levels. CPT-induced Tra2 down-regulation was ATR-dependent and temporally paralleled the change in TAF1 alternative splicing, supporting the conclusion that Tra2 directly regulates TAF1 alternative splicing. Additionally, CPT-induced Tra2 down-regulation occurred independently of new protein synthesis, suggesting a post-translational mechanism. The proteasome inhibitor MG132 reduced CPT-induced Tra2 degradation and TAF1 alternative splicing, and mutation of evolutionarily conserved Tra2 lysine 81, a potential ubiquitin conjugation site, to arginine inhibited CPT-induced Tra2 degradation, supporting a proteasome dependent alternative splicing mechanism. We conclude that CPT-induced TAF1 alternative splicing occurs through ATR-signaled degradation of a subset of splicing-regulatory proteins. PMID- 19218246 TI - 14-3-3:Shc scaffolds integrate phosphoserine and phosphotyrosine signaling to regulate phosphatidylinositol 3-kinase activation and cell survival. AB - Integrated cascades of protein tyrosine and serine/threonine phosphorylation play essential roles in transducing signals in response to growth factors and cytokines. How adaptor or scaffold proteins assemble signaling complexes through both phosphotyrosine and phosphoserine/threonine residues to regulate specific signaling pathways and biological responses is unclear. We show in multiple cell types that endogenous 14-3-3zeta is phosphorylated on Tyr(179) in response to granulocyte macrophage colony-stimulating factor. Importantly, 14-3-3zeta can function as an intermolecular bridge that couples to phosphoserine residues and also directly binds the SH2 domain of Shc via Tyr(179). The assembly of these 14 3-3:Shc scaffolds is specifically required for the recruitment of a phosphatidylinositol 3-kinase signaling complex and the regulation of CTL-EN cell survival in response to cytokine. The biological significance of these findings was further demonstrated using primary bone marrow-derived mast cells from 14-3 3zeta(-/-) mice. We show that cytokine was able to promote Akt phosphorylation and viability of primary mast cells derived from 14-3-3zeta(-/-) mice when reconstituted with wild type 14-3-3zeta, but the Akt phosphorylation and survival response was reduced in cells reconstituted with the Y179F mutant. Together, these results show that 14-3-3:Shc scaffolds can act as multivalent signaling nodes for the integration of both phosphoserine/threonine and phosphotyrosine pathways to regulate specific cellular responses. PMID- 19218247 TI - Human cytosolic hydroxysteroid dehydrogenases of the aldo-ketoreductase superfamily catalyze reduction of conjugated steroids: implications for phase I and phase II steroid hormone metabolism. AB - Aldo-ketoreductase 1C (AKR1C) enzymes catalyze the NADPH-dependent reduction of ketosteroids to hydroxysteroids. They are Phase I metabolizing enzymes for natural and synthetic steroid hormones. They convert 5alpha-dihydrotestosterone (Dht, potent androgen) to 3alpha/beta-androstanediols (inactive androgens) and the prodrug tibolone (Tib) to estrogenic 3alpha/beta-hydroxytibolones. Herein we demonstrate for the first time that human AKR1C enzymes (AKR1C1-4) are able to reduce conjugated steroids such as Dht-17beta-glucuronide (DhtG), Dht-17beta sulfate (DhtS), and Tib-17beta-sulfate (TibS). Product identities were characterized by liquid chromatography-mass spectrometry, and kinetic parameters of the reactions were determined. The product profile of the reduction of each steroid conjugate by the individual AKR1C isoform was similar to that of the corresponding free steroid except for the reduction of DhtG catalyzed by AKR1C2, where a complete inversion in stereochemical preference to 3beta-reduction (with DhtG) from 3alpha-reduction (with Dht and DhtS) was observed. The catalytic efficiency of 3-keto reduction was modestly affected by the presence of a 17 sulfate group but severely impaired by the presence of a 17-glucuronide group for AKR1C1-3 isoforms. AKR1C4, however, showed superior catalytic efficiencies versus the other isoforms, and those were unaffected by steroid conjugation. Our findings provide evidence for alternative pathways of steroid metabolism where the phase I reaction (reduction) occurs after the phase II reaction (conjugation). Specifically, it is indicated that Dht is metabolized to its metabolite 3alpha-androstanediol-17-glucuronide via the previously unrecognized "conjugation pathway" involving the sequential reactions of UGT2B17 and AKR1C4 in liver but via the conventional "reduction pathway" involving the sequential reactions of AKR1C2 and UGT2B15/17 in prostate. PMID- 19218248 TI - Cyclooxygenase Allosterism, Fatty Acid-mediated Cross-talk between Monomers of Cyclooxygenase Homodimers. AB - Prostaglandin endoperoxide H synthases (PGHSs) 1 and 2, also known as cyclooxygenases (COXs), catalyze the oxygenation of arachidonic acid (AA) in the committed step in prostaglandin (PG) biosynthesis. PGHSs are homodimers that display half of sites COX activity with AA; thus, PGHSs function as conformational heterodimers. Here we show that, during catalysis, fatty acids (FAs) are bound at both COX sites of a PGHS-2 dimer. Initially, an FA binds with high affinity to one COX site of an unoccupied homodimer. This monomer becomes an allosteric monomer, and it causes the partner monomer to become the catalytic monomer that oxygenates AA. A variety of FAs can bind with high affinity to the COX site of the monomer that becomes the allosteric monomer. Importantly, the efficiency of AA oxygenation is determined by the nature of the FA bound to the allosteric monomer. When tested with low concentrations of saturated and monounsaturated FAs (e.g. oleic acid), the rates of AA oxygenation are typically 1.5-2 times higher with PGHS-2 than with PGHS-1. These different kinetic behaviors of PGHSs may account for the ability of PGHS-2 but not PGHS-1 to efficiently oxygenate AA in intact cells when AA is a small fraction of the FA pool such as during "late phase" PG synthesis. PMID- 19218250 TI - Randomised controlled trial of posterior sub-Tenon triamcinolone as adjunct to panretinal photocoagulation for treatment of diabetic retinopathy. AB - AIMS: To evaluate the efficacy of a single posterior sub-Tenon capsule injection of triamcinolone acetonide (PSTA) before panretinal photocoagulation (PRP). METHODS: This 6-month study involved the randomisation of 82 eyes of 41 patients, with bilateral severe non-proliferative diabetic retinopathy or proliferative diabetic retinopathy to a single PSTA 20 mg or to no injection before PRP. The primary end-point was change in best-corrected visual acuity (BCVA) at 6 months compared with that at baseline using the logarithm of the minimum angle of resolution (logMAR). Secondary end-points were changes in retinal thickness and intraocular pressure. RESULTS: The mean changes in logMAR BCVA at 6 months compared with that at baseline were a worsening of 0.010 (SD 0.029) in the control group (no injection) and an improvement of 0.072 (0.028) in the PSTA group (p = 0.04). The mean changes in foveal thickness at 6 months compared with baseline measurements were an increase of 32.8 (82.8) mum in the control group and a lessening of 9.7 (85.6) mum in the PSTA group (p = 0.03). CONCLUSIONS: PSTA before PRP appears to be beneficial in preventing PRP-induced visual loss in eyes with diabetic retinopathy by reducing the chance of macular thickening. PMID- 19218249 TI - Nadolol plus isosorbide mononitrate alone or associated with band ligation in the prevention of recurrent bleeding: a multicentre randomised controlled trial. AB - BACKGROUND AND AIMS: Previous clinical trials suggest that adding non-selective beta-blockers improves the efficacy of endoscopic band ligation (EBL) in the prevention of recurrent bleeding, but no study has evaluated whether EBL improves the efficacy of beta-blockers + isosorbide-5-mononitrate. The present study was aimed at evaluating this issue in a multicentre randomised controlled trial (RCT) and to correlate changes in hepatic venous pressure gradient (HVPG) during treatment with clinical outcomes METHODS: 158 patients with cirrhosis, admitted because of variceal bleeding, were randomised to receive nadolol+isosorbide-5 mononitrate alone (Drug: n = 78) or combined with EBL (Drug+EBL; n = 80). HVPG measurements were performed at randomisation and after 4-6 weeks on medical therapy. RESULTS: Median follow-up was 15 months. One-year probability of recurrent bleeding was similar in both groups (33% vs 26%: p = 0.3). There were no significant differences in survival or need of rescue shunts. Overall adverse events or those requiring hospital admission were significantly more frequent in the Drug+EBL group. Recurrent bleeding was significantly more frequent in HVPG non-responders than in responders (HVPG reduction >or=20% or 0.05). There was a significant time effect for large arterial compliance (P < 0.05), which increased in both groups. CONCLUSIONS: In contrast to previous studies in men, which found decrease in arterial compliance with resistance training, no decrease in arterial compliance was observed. PMID- 19218271 TI - Pharmacokinetics of caspofungin and voriconazole in critically ill patients during extracorporeal membrane oxygenation. AB - OBJECTIVES: During extracorporeal membrane oxygenation (ECMO), drug disposition changes significantly. Plasma concentrations are altered due to an expanded circulating volume leading to a decreased elimination. In addition, adsorption and sequestration of drugs by the ECMO circuit components may further alter pharmacokinetics. Treating patients during the ECMO period with antifungals is difficult. Loss in the ECMO circuit can potentially result in sub-therapeutic levels. METHODS: Two cases are presented in which caspofungin and voriconazole levels and pharmacokinetic parameters were determined during the ECMO period. RESULTS: Mean caspofungin trough and peak levels were 3.73 and 11.95 microg/mL. These are comparable to previously reported ones. Also pharmacokinetic parameters were identical to those reported in the literature. It seems that caspofungin is not sequestrated by the ECMO circuit, which is expected based on its low log P value. During the first days of ECMO therapy, voriconazole trough and peak levels did not differ much from those determined prior to ECMO therapy. However, at the start of ECMO therapy, the voriconazole dose was increased from 280 to 400 mg twice daily as loss due to binding to the circuit was expected. This increase was not immediately reflected in higher voriconazole levels, which may be due to drug sequestration by the circuit. However, the voriconazole half-life was extended up to 20 h in our patient. Two days after the dose increase, levels reached troughs >10 microg/mL and peaks of around 15 microg/mL, exceeding the therapeutic interval for voriconazole. This can possibly be explained by the saturation of binding sites on the ECMO circuit. CONCLUSIONS: Our results suggest that adequate caspofungin plasma levels are maintained during ECMO. In the case of voriconazole, it is recommended to monitor plasma levels to ensure efficacy and avoid toxicity. PMID- 19218273 TI - Prescription patterns of antimicrobials in veterinary practices in Switzerland. AB - OBJECTIVES: The objective of this study was to analyse antimicrobial prescriptions by veterinarians and to evaluate the appropriateness of use compared with prudent use guidelines. PRACTICES AND METHODS: Computerized records of prescriptions and treatments from eight mixed veterinary practices were analysed over a period of 2 years. A total of 61 212 antimicrobial treatments were recorded. Treatments were classified according to animal species treated, indication for treatment, route of administration and antimicrobial class used. For each treatment and antimicrobial substance, the prescribed dose was calculated. Dosage, antimicrobial classes and combinations of different classes used for different indications were compared with published recommendations. RESULTS: From the total amount of 1590 kg of active antimicrobial substance, sulphonamides (594 kg), tetracyclines (335 kg), and penicillins and cephalosporins (290 kg) were the classes of which the largest quantity was prescribed. Penicillins and cephalosporins were most frequently prescribed (37% of treatments), followed by aminoglycosides (18%), tetracyclines (14%) and sulphonamides (11%). Sixty-one per cent of the amount of antimicrobials prescribed was used for the treatment of groups of animals via feed or water. Antimicrobial classes classified as highest priority for human medicine by an international group of experts were used in 9% of the prescriptions. The dosage corresponded to the manufacturer's recommendation in 45% of the analysed prescriptions. CONCLUSIONS: Most prescriptions corresponded well to guidelines on prudent use of antimicrobials. Nevertheless, the large variation of prescriptions among different veterinarians indicates that the usage of critical antimicrobial substances and the amount of antimicrobials used for group medication without a specific indication could be further reduced. PMID- 19218272 TI - Evaluation of the lymphocyte trafficking drug FTY720 in SHIVSF162P3-infected rhesus macaques. AB - OBJECTIVES: FTY720 causes retention of lymphocytes in lymphatic tissues. Previous studies revealed that FTY720 can decrease or eliminate chronic viral infections of mice. We address here whether therapeutic use of FTY720 in simian human immunodeficiency virus (SHIV)-infected rhesus macaques could also decrease viraemia. METHODS: FTY720 was administered intravenously to three SHIV(SF162P3) infected macaques at 39, 7 or 6 weeks of infection; three control macaques (47, 48 or 6 weeks of infection) did not receive drug. FTY720 was given at 0.004 mg/kg on days 0, 1, 2, 14, 15 and 16, followed by 0.1 mg/kg on days 28, 29, 30, 42, 43 and 44. Blood was collected seven times throughout and four times during 47 days of follow-up. RESULTS: Only the 0.1 mg/kg dose resulted in a reduction in mean blood CD4+ T cells and B cells (to 33% and 27% of pre-drug levels, P=0.0024 and 0.003, respectively). FTY720 treatment did not lead to significant deviations from the natural pattern of viral control. Plasma viraemia progressed from a range of 10(4)-10(2) copies/mL before treatment to 10(4)-temporarily undetectable levels on the last day of treatment. SHIV(SF162P3) was not eliminated, however, as plasma viraemia and proviral DNA persisted during the follow-up. No significant alterations in T cell activity were noted throughout the drug course. CONCLUSIONS: FTY720 administration had no detectable therapeutic effect at the doses and schedules outlined here, although blood CD4+ T cells and B cells were effectively reduced. Future work might reveal whether FTY720 could be beneficial in more pathogenic SHIV, simian immunodeficiency virus or HIV infections. PMID- 19218274 TI - Therapeutic impact of the correlation of doxycycline serum concentrations and the decline of phase I antibodies in Q fever endocarditis. AB - BACKGROUND: Endocarditis is the main clinical manifestation of chronic Q fever, a challenging disease due to Coxiella burnetii. The recommended treatment for Q fever endocarditis is a combination of doxycycline and hydroxychloroquine for at least 18 months. However, there is considerable heterogeneity in the biological response to this regimen as measured by the rate of decrease of dilutions of phase I antibodies against C. burnetii. Previous studies have demonstrated that this response heterogeneity was due to variations in the serum concentration of doxycycline in patients when compared with MICs for the isolates. OBJECTIVES: The objective of this study was to evaluate retrospectively the evolution of phase I antibodies in patients with an initial slow serological change, who received higher doses of doxycycline. RESULTS: Among 17 patients with definitive diagnosis of Q fever endocarditis, 12 were defined as slow responders [mean (+/- SD) decrease of dilutions of phase I antibodies of 0.42 +/- 0.51 and mean (+/-SD) serum doxycycline level of 3.44 +/- 1.06 mg/L after 1 year of treatment] and received increased doses of doxycycline. After 1 year of treatment with increased dose, the mean (+/-SD) serum doxycycline concentration increased to 4.86 +/- 1.14 mg/L (P<0.05) and the mean (+/-SD) decrease of dilutions of phase I antibodies increased to 3.42 +/- 1.78 (P<0.05). CONCLUSIONS: During the treatment of Q fever endocarditis, serum concentrations of doxycycline should be monitored concomitantly with phase I antibodies in order to adjust the dose of doxycycline to achieve a higher concentration for patients with slow serological evolution. PMID- 19218275 TI - In vitro selection of resistance in Streptococcus pneumoniae at in vivo fluoroquinolone concentrations. AB - OBJECTIVES: To compare the ability to select for resistance in Streptococcus pneumoniae of levofloxacin, moxifloxacin, ciprofloxacin and prulifloxacin. METHODS: Twenty strains of S. pneumoniae susceptible to fluoroquinolones were used. The frequencies of spontaneous single-step mutations at plasma and epithelial lining fluid (ELF) peak and trough antibiotic concentrations were calculated. Multi-step selection of resistance was evaluated by performing 10 serial subcultures on agar plates containing a linear gradient from peak to trough antimicrobial concentrations, followed by 10 subcultures on antibiotic free agar. Resistant strains selected after multi-step selection were characterized for DNA mutations by sequencing gyrA, gyrB, parC and parE genes. RESULTS: Levofloxacin and moxifloxacin showed the lowest frequencies of mutations (median <10(-11)) at plasma peak and at ELF concentrations, while medians ranging from 10(-8) to 10(-6) were observed for ciprofloxacin and prulifloxacin. In a multi-step selection assay, ciprofloxacin and prulifloxacin selected for the highest number of resistant strains (19 and 31, respectively). No selection of resistance was observed for levofloxacin at ELF concentrations and for moxifloxacin at plasma and ELF concentrations. Mutations in parC, parE and gyrA genes were found in ciprofloxacin- and prulifloxacin-resistant strains, while only parC mutations were found for levofloxacin. CONCLUSIONS: Levofloxacin and moxifloxacin are characterized by a lower propensity to select in vitro for resistance in S. pneumoniae than ciprofloxacin and prulifloxacin, when tested at plasma and lung concentrations. PMID- 19218276 TI - TR-700 in vitro activity against and resistance mutation frequencies among Gram positive pathogens. AB - BACKGROUND: TR-700, the active component of the oxazolidinone prodrug TR-701, has demonstrated potent activity against numerous Gram-positive species. In this study, single-step mutation frequencies, passaging and the activity of TR-700 were tested against a worldwide collection of linezolid-non-susceptible organisms and matched controls. METHODS: One hundred and twenty linezolid-non-susceptible and 120 controls matched by genus/species, geographic origin, site of infection and time were susceptibility tested by reference broth microdilution methods. Species of isolates were: Enterococcus faecalis (16 linezolid non-susceptible/16 wild-type strains); Enterococcus faecium (55/55), Staphylococcus aureus (8/8); coagulase-negative staphylococci (at least 7 spp., 40/40) and viridans group streptococci (2 spp., 1/1). 23S rRNA target mutations or cfr genes were detected by PCR and sequencing. RESULTS: Among linezolid-non-susceptible strains, the resistance mechanisms were G2576T (109), cfr (4) and unknown (7), with strains originating from Europe, Far East and North and South America. Most strains were multidrug-resistant and cfr isolates exhibited co-resistance to phenicols, clindamycin, linezolid, pleuromutilins and streptogramin B. TR-700 MIC values, regardless of species, were 4-32-fold lower than those of linezolid. TR-700 MIC results were < or = 4, < or = 8 or < or = 16 mg/L for 88%, 96% and > 99% of linezolid-non-susceptible strains, respectively. Spontaneous single-step mutations were undetected (<1.1 x 10(-9)) and 14 day passaging studies produced modest TR-700 MIC elevations compared with linezolid controls. CONCLUSIONS: TR 700 exhibited enhanced activity against linezolid-non-susceptible and wild-type control strains of Gram-positive cocci. A significant number (nearly 90%) of linezolid-non-susceptible strains were inhibited by potentially achievable levels (< or = 4 mg/L) of TR-700. All strains with the emerging cfr-mediated resistance determinant had TR-700 MIC results at < or = 8 mg/L. PMID- 19218277 TI - Identification of the qnr family in Enterobacteriaceae in clinical practice. PMID- 19218278 TI - Effects of iron depletion on Entamoeba histolytica alcohol dehydrogenase 2 (EhADH2) and trophozoite growth: implications for antiamoebic therapy. AB - OBJECTIVES: The purpose of this study was to determine the mechanism by which iron chelation affects the trophozoite survival of Entamoeba histolytica. Fe2+ is a cofactor for E. histolytica alcohol dehydrogenase 2 (EhADH2), an essential bifunctional enzyme [alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH)] in the glycolytic pathway of E. histolytica. METHODS: We tested the effects of iron depletion on trophozoite growth, the kinetics of iron binding to EhADH2, and the activities of ADH and ALDH. RESULTS: Growth of E. histolytica trophozoites, and ADH and ALDH enzymatic activities were directly inhibited by iron chelation. Kinetics of iron binding to EhADH2 reveals the differential iron affinity of ADH (higher) and ALDH (lower). CONCLUSIONS: This study demonstrates that iron chelation interrupts the completion of the fermentative pathway of E. histolytica by removing the metal cofactor indispensable for the structural and functional stability of EhADH2, thus affecting trophozoite survival. We propose that iron-starvation-based strategies could be used to treat amoebiasis. PMID- 19218279 TI - New therapeutic advances in the management of progressive thyroid cancer. AB - The spectrum of thyroid cancers ranges from one of the most indolent to one of the most aggressive solid tumors identified. Conventional therapies for thyroid cancers are based on the histologic type of thyroid cancers such as papillary or follicular thyroid cancer (differentiated thyroid cancer (DTC)), medullary thyroid cancer (MTC), or anaplastic thyroid cancer (ATC). While surgery is one of the key treatments for all such types of thyroid cancers, additional therapies vary. Effective targeted therapy for DTC is a decades-old practice with systemic therapies of thyroid stimulating hormone suppression and radioactive iodine therapy. However, for the iodine-refractory DTC, MTC, and ATC there is no effective systemic standard of care treatment. Recent advances in understanding pathogenesis of DTC and development of molecular targeted therapy have dramatically transformed the field of clinical research in thyroid cancer. Over the last five years, incredible progress has been made and phases I-III clinical trials have been conducted in various types of thyroid cancers with some remarkable results that has made an impact on lives of patients with thyroid cancer. Such history-making events have boosted enthusiasm and interest among researchers, clinicians, patients, and sponsors and we anticipate ongoing efforts to develop more effective and safe therapies for thyroid cancer. PMID- 19218280 TI - Loss of neuronatin expression is associated with promoter hypermethylation in pituitary adenoma. AB - The imprinted gene, neuronatin (NNAT), is one of the most abundant transcripts in the pituitary and is thought to be involved in the development and maturation of this gland. In a recent whole-genome approach, exploiting a pituitary tumour cell line, we identified hypermethylation associated loss of NNAT. In this report, we determined the expression pattern of NNAT in individual cell types of the normal gland and within each of the different pituitary adenoma subtypes. In addition, we determined associations between expression and CpG island methylation and used colony forming efficiency assays (CFE) to gain further insight into the tumour suppressor function of this gene. Immunohistochemical (IHC) co-localization studies of normal pituitaries showed that each of the hormone secreting cells (GH, PRL, ACTH, FSH and TSH) expressed NNAT. However, 33 out of 47 adenomas comprising, 11 somatotrophinomas, 10 prolactinomas, 12 corticotrophinomas and 14 non-functioning tumours, irrespective of subtype failed to express either NNAT transcript or protein as determined by quantitative real-time RT-PCR and IHC respectively. In normal pituitaries and adenomas that expressed NNAT the promoter associated CpG island showed characteristics of an imprinted gene where approximately 50% of molecules were densely methylated. However, in the majority of adenomas that showed loss or significantly reduced expression of NNAT, relative to normal pituitaries, the gene-associated CpG island showed significantly increased methylation. Induced expression of NNAT in transfected AtT-20 cells significantly reduced CFE. Collectively, these findings point to an important role for NNAT in the pituitary and perhaps tumour development in this gland. PMID- 19218281 TI - Microarray gene expression and immunohistochemistry analyses of adrenocortical tumors identify IGF2 and Ki-67 as useful in differentiating carcinomas from adenomas. AB - The management of adrenocortical tumors (ACTs) is complex. The Weiss score is the present most widely used system for ACT diagnosis. An ACT is scored from 0 to 9, with a higher score correlating with increased malignancy. However, ACTs with a score of 3 can be phenotypically benign or malignant. Our objective is to use microarray profiling of a cohort of adrenocortical carcinomas (ACCs) and adrenocortical adenomas (ACAs) to identify discriminatory genes that could be used as an adjunct to the Weiss score. A cohort of Weiss score defined ACCs and ACAs were profiled using Affymetrix HGU133plus2.0 genechips. Genes with high discriminatory power were identified by univariate and multivariate analyses and confirmed by quantitative real-time reverse transcription PCR and immunohistochemistry (IHC). The expression of IGF2, MAD2L1, and CCNB1 were significantly higher in ACCs compared with ACAs while ABLIM1, NAV3, SEPT4, and RPRM were significantly lower. Several proteins, including IGF2, MAD2L1, CCNB1, and Ki-67 had high-diagnostic accuracy in differentiating ACCs from ACAs. The best results, however, were obtained with a combination of IGF2 and Ki-67, with 96% sensitivity and 100% specificity in diagnosing ACCs. Microarray gene expression profiling accurately differentiates ACCs from ACAs. The combination of IGF2 and Ki-67 IHC is also highly accurate in distinguishing between the two groups and is particularly helpful in ACTs with Weiss score of 3. PMID- 19218282 TI - Differential retention of metabolic genes following whole-genome duplication. AB - Classical studies in Metabolic Control Theory have shown that metabolic fluxes usually exhibit little sensitivity to changes in individual enzyme activity, yet remain sensitive to global changes of all enzymes in a pathway. Therefore, little selective pressure is expected on the dosage or expression of individual metabolic genes, yet entire pathways should still be constrained. However, a direct estimate of this selective pressure had not been evaluated. Whole-genome duplications (WGDs) offer a good opportunity to address this question by analyzing the fates of metabolic genes during the massive gene losses that follow. Here, we take advantage of the successive rounds of WGD that occurred in the Paramecium lineage. We show that metabolic genes exhibit different gene retention patterns than nonmetabolic genes. Contrary to what was expected for individual genes, metabolic genes appeared more retained than other genes after the recent WGD, which was best explained by selection for gene expression operating on entire pathways. Metabolic genes also tend to be less retained when present at high copy number before WGD, contrary to other genes that show a positive correlation between gene retention and preduplication copy number. This is rationalized on the basis of the classical concave relationship relating metabolic fluxes with enzyme expression. PMID- 19218283 TI - Does having a twin brother make for a bigger brain? AB - OBJECTIVE: Brain volume of boys is larger than that of girls by approximately 10%. Prenatal exposure to testosterone has been suggested in the masculinization of the brain. For example, in litter-bearing mammals intrauterine position increases prenatal testosterone exposure through adjacent male fetuses, resulting in masculinization of brain morphology. DESIGN: The influence of intrauterine presence of a male co-twin on masculinization of human brain volume was studied in 9-year old twins. METHODS: Magnetic resonance imaging brain scans, current testosterone, and estradiol levels were acquired from four groups of dizygotic (DZ) twins: boys from same-sex twin-pairs (SSM), boys from opposite-sex twin pairs (OSM), girls from opposite-sex twin-pairs (OSF), and girls from same-sex twin-pairs (SSF; n=119 individuals). Data on total brain, cerebellum, gray and white matter volumes were examined. RESULTS: Irrespective of their own sex, children with a male co-twin as compared to children with a female co-twin had larger total brain (+2.5%) and cerebellum (+5.5%) volumes. SSM, purportedly exposed to the highest prenatal testosterone levels, were found to have the largest volumes, followed by OSM, OSF and SSF children. Birth weight partly explained the effect on brain volumes. Current testosterone and estradiol levels did not account for the volumetric brain differences. However, the effects observed in children did not replicate in adult twins. CONCLUSIONS: Our study indicates that sharing the uterus with a DZ twin brother increases total brain volume in 9-year olds. The effect may be transient and limited to a critical period in childhood. PMID- 19218284 TI - Germ cell sex determination in mammals. AB - One of the major decisions that germ cells make during their development is whether to differentiate into oocytes or sperm. In mice, the germ cells' decision to develop as male or female depends on sex-determining signalling molecules in the embryonic gonadal environment rather than the sex chromosome constitution of the germ cells themselves. In response to these sex-determining cues, germ cells in female embryos initiate oogenesis and enter meiosis, whereas germ cells in male embryos initiate spermatogenesis and inhibit meiosis until after birth. However, it is not clear whether the signalling molecules that mediate germ cell sex determination act in the developing testis or the developing ovary, or what these signalling molecules might be. Here, we review the evidence for the existence of meiosis-inducing and meiosis-preventing substances in the developing gonad, and more recent studies aimed at identifying these molecules in mice. In addition, we discuss the possibility that some of the reported effects of these factors on germ cell development may be indirect consequences of impairing sexual differentiation of gonadal somatic cells or germ cell survival. Understanding the molecular mechanisms of germ cell sex determination may provide candidate genes for susceptibility to germ cell tumours and infertility in humans. PMID- 19218285 TI - Misexpression of CCAAT/enhancer binding protein beta causes osteopenia. AB - CCAAT/enhancer binding proteins (C/EBPs) are expressed by osteoblasts and adipocytes during differentiation. C/EBP beta is critical for adipogenesis; however, its role in osteoblastogenesis is unclear, and its function in the postnatal skeleton is not known. To study C/EBP beta in osteoblasts in vivo, we created transgenic mice expressing full length C/EBP beta under the control of a 3.8 kb fragment of the human osteocalcin promoter. Two transgenic lines were established in a friend leukemia virus strain B genetic background, and compared with wild type littermate controls. Both C/EBP beta transgenic lines exhibited osteopenia, with a 30% decrease in bone volume, due to a decrease in trabecular number. The number of osteoblasts and osteoclasts per bone perimeter was not changed. Bone marrow stromal cells from C/EBP beta transgenics showed reduced mineralization, and reduced alkaline phosphatase mRNA levels. Calvarial osteoblasts from C/EBP beta transgenics displayed reduced alkaline phosphatase activity. To determine the consequences of the Cebpb deletion in vivo, the phenotype of Cebpb null mice was compared with that of wild type controls of identical genetic composition. Cebpb null mice exhibited reduced weight, body fat, and bone mineral density, and decreased bone volume, due to a decrease in trabecular number. The number of osteoblasts and osteoclasts per bone perimeter was not changed. C/EBP beta downregulation by RNA interference in calvarial osteoblasts had no effect on osteoblast differentiation/function. The phenotype of the Cebpb inactivation may be secondary to systemic indirect effects, and to direct effects of C/EBP beta in osteoblasts. In conclusion, C/EBP beta plays a role in mesenchymal cell differentiation and its misexpression in vivo causes osteopenia. PMID- 19218286 TI - Cardioprotective growth factors. AB - Many of the originally identified cardiovascular 'growth factors' have been demonstrated to exert a diverse variety of actions within the cardiovascular system, the majority of which are unrelated to their initially proposed mechanism of action. Interestingly, several of these growth factors have been demonstrated to protect the cardiomyocyte from the detrimental effects of acute ischaemia reperfusion injury, through the activation of a variety of cell-surface receptors and the subsequent recruitment of a number of intracellular signal transduction pathways, which include components of the reperfusion injury salvage kinase pathway. This article will review several of these cardioprotective growth factors with respect to their ability to confer direct myocardial protection, focusing on the underlying signalling pathways involved and their potential for clinical application. PMID- 19218287 TI - Prominent role of KCa3.1 in endothelium-derived hyperpolarizing factor-type dilations and conducted responses in the microcirculation in vivo. AB - AIMS: The activation of endothelial Ca2+-dependent K+-channels, KCa3.1 (IKCa), and KCa2.3 (SKCa) has been proposed to be a prerequisite for endothelial hyperpolarization, which subsequently hyperpolarizes and relaxes smooth muscle [endothelium-derived hyperpolarizing factor (EDHF)-type dilation] and initiates conducted dilations. Although EDHF is the main mediator of acetylcholine (ACh) induced dilation in the murine skeletal microcirculation, the differential contribution of KCa3.1 and KCa2.3 is not known. METHODS AND RESULTS: We assessed agonist-induced and conducted dilations as well as endothelial hyperpolarization in the cremaster microcirculation of KCa3.1-deficient (KCa3.1-/-) and wild-type mice (wt) in vivo after blockade of NO and prostaglandins. Compared with wt, resting tone was enhanced by approximately 25% in arterioles of KCa3.1-/- mice. ACh-induced dilations in KCa3.1-/- mice were virtually abolished at low and intermediate concentrations and a remaining dilation at 10 micromol/L ACh was abrogated by blockade of KCa2.3 with UCL1684. Sodium nitroprusside- and adenosine induced dilations were similar in wt and KCa3.1-/-. Focal application of ACh induced dilations at the local site in both genotypes, which conducted along the vessel. However, the amplitude of the dilation decreased with distance only in KCa3.1-/-. Blockade of KCa2.3 in wt did not affect conducted dilations. A KCa3.1 opener induced a conducting dilation in wt but not in KCa3.1-/-. Membrane potential recordings in vivo demonstrated endothelial hyperpolarization in response to ACh in both genotypes; however, the hyperpolarization was severely impaired in KCa3.1-/- (Delta membrane potential: -3 +/- 1 vs. -14 +/- 2 mV). CONCLUSION: We conclude that KCa3.1 is of major importance for endothelial hyperpolarization and EDHF-type responses in skeletal muscle arterioles, and its deficiency is not compensated by KCa2.3. Sole activation of KCa3.1 is capable of initiating conducted responses, and KCa3.1 may contribute to the propagation of the signal, although its presence is not mandatory. PMID- 19218288 TI - Comparison of contractile mechanisms of sphingosylphosphorylcholine and sphingosine-1-phosphate in rabbit coronary artery. AB - AIMS: Although stimulation with sphingosylphosphorylcholine (SPC) or sphingosine 1-phosphate (S1P) generally leads to similar vascular responses, the contractile patterns and their underlying signalling mechanisms are often distinct. We investigated the different reliance upon Ca2+-dependent and Ca2+-sensitizing mechanisms of constriction in response to SPC or S1P in coronary arteries. METHODS AND RESULTS: Contractile responses, changes in [Ca2+]i, and phosphorylation of myosin light chain phosphatase-targeting subunit (MYPT1) were measured. SPC induced a concentration-dependent sustained contraction. S1P evoked a rapid rise in force (initial transient), which was followed by a secondary sustained force. In the absence of extracellular Ca2+, the concentration dependency of constriction to SPC was shifted to the right, but with no change in maximum force, whereas S1P-induced contraction was significantly blunted. Cyclopiazonic acid (CPA) significantly decreased the initial transient force induced by S1P. In isolated single cells, S1P markedly increased [Ca2+]i, whereas only a modest elevation was noted with SPC. The S1P-induced elevation of [Ca2+]i was abolished by pre-treatment with CPA and was significantly reduced in the absence of extracellular Ca2+. In beta-escin-permeabilized strips, SPC augmented pCa 6.3-induced force; this was significantly inhibited by fasudil hydrochloride. S1P induced little or no augmentation of pCa 6.3-induced force. In intact arteries, SPC-induced contraction was completely inhibited by fasudil hydrochloride. Fasudil hydrochloride had no effect on the initial transient force induced by S1P but significantly inhibited the secondary sustained force. SPC induced a several-fold increase in Thr696 and Thr853 phosphorylation of MYPT1, but S1P did not affect phosphorylation of MYPT1. CONCLUSION: Our results suggest that constriction of coronary arteries in response to the bioactive lipid S1P or SPC occurs by distinct signalling pathways. Activation of the RhoA/RhoA associated kinase pathway and subsequent phosphorylation of MYPT1 play a key role in SPC-induced coronary contraction, whereas elevation of [Ca2+]i is crucial for S1P-induced coronary constriction. PMID- 19218289 TI - Prevention of cardiomyopathy in delta-sarcoglycan knockout mice after systemic transfer of targeted adeno-associated viral vectors. AB - AIMS: Delta-sarcoglycan is a member of the dystrophin-associated glycoprotein complex linking the cytoskeleton to the extracellular matrix. Similar to patients with defects in the gene encoding delta-sarcoglycan (Sgcd), knockout mice develop cardiomyopathy and muscular dystrophy. The aim of our study was to develop an approach for preventing cardiomyopathy in Sgcd-deficient mice by cardiac expression of the intact cDNA upon systemic delivery of adeno-associated viral (AAV) vectors. METHODS AND RESULTS: We packaged the Sgcd cDNA under transcriptional control of a myosin light chain-promoter fused with a cytomegalovirus enhancer into AAV-9 capsids. Vectors carrying either the Sgcd cDNA or an enhanced green fluorescent protein (EGFP) reporter gene were intravenously injected into adult Sgcd knockout mice. After 6 months, immunohistochemistry revealed almost complete reconstitution of the sarcoglycan subcomplex in heart but not skeletal muscle of mice with the Sgcd vector. Furthermore, Sgcd gene transfer resulted in prevention of cardiac fibrosis and significantly increased running distance measured by voluntary wheel running. Left ventricular function remained stable in mice expressing Sgcd while it deteriorated in EGFP controls within 6 months, paralleled by increased expression of brain natriuretic peptide, a molecular marker of heart failure. CONCLUSION: Our study establishes an approach to specifically treat hereditary cardiomyopathies by targeting gene expression into the myocardium upon systemic application of AAV vectors. PMID- 19218295 TI - Cochrane review: information provision for stroke patients and their caregivers. AB - OBJECTIVE: To assess the effectiveness of information provision strategies in improving the outcome for stroke patients and/or their identified caregivers. DATA SOURCES: We searched: the Cochrane Stroke Group Trials Register; the Cochrane Central Register of Controlled Trials; electronic databases MEDLINE; EMBASE; CINAHL; PsycINFO; Science Citation Index and Social Science Citation Index; Assia; Index to UK theses; Dissertation Abstracts; ongoing trials and research registers; bibliographies of retrieved papers, relevant articles, and books; the Journal of Advanced Nursing. We also contacted researchers for additional information. REVIEW METHODS: Two review authors independently assessed trial eligibility, extracted data and assessed methodological quality. Primary outcomes were knowledge about stroke and impact on mood. Meta-analyses were undertaken for the domains of knowledge, mood, satisfaction, and mortality. RESULTS: Seventeen trials were identified and 11 contributed data to the meta analyses. There were significant effects in favour of the intervention on patient knowledge (standardized mean difference (SMD) 0.29, 95% confidence interval (CI) 0.12 to 0.46), caregiver knowledge (SMD 0.74 95% CI 0.06 to 1.43), patient depression scores (weighted mean difference (WMD) -0.52, 95% CI -0.93 to -0.10), and one aspect of patient satisfaction (odds ratio (OR) 2.07, 95% CI 1.33 to 3.23). Post-hoc subgroup analyses showed that strategies which actively involved patient and caregivers had a significantly greater effect on patient anxiety (P<0.05) and depression (P<0.02) than passive strategies. CONCLUSION: There is some evidence to support the routine provision of information to stroke patients and their families. Although the best way to provide information is still not clear, the results of this review suggest that strategies which actively involve patients and caregivers should be used in routine practice. PMID- 19218294 TI - Modification of the effect of vitamin E supplementation on the mortality of male smokers by age and dietary vitamin C. AB - The Alpha-Tocopherol, Beta-Carotene Cancer Prevention (ATBC) Study (1985-1993) recruited 29,133 Finnish male cigarette smokers, finding that vitamin E supplementation had no overall effect on mortality. The authors of this paper found that the effect of vitamin E on respiratory infections in ATBC Study participants was modified by age, smoking, and dietary vitamin C intake; therefore, they examined whether the effect of vitamin E supplementation on mortality is modified by the same variables. During a median follow-up time of 6.1 years, 3,571 deaths occurred. Age and dietary vitamin C intake had a second order interaction with vitamin E supplementation of 50 mg/day. Among participants with a dietary vitamin C intake above the median of 90 mg/day, vitamin E increased mortality among those aged 50-62 years by 19% (95% confidence interval: 5, 35), whereas vitamin E decreased mortality among those aged 66-69 years by 41% (95% CI: -56, -21). Vitamin E had no effect on participants who had a dietary vitamin C intake below the median. Smoking quantity did not modify the effect of vitamin E. This study provides strong evidence that the effect of vitamin E supplementation on mortality varies between different population groups. Further study is needed to confirm this heterogeneity. PMID- 19218296 TI - Effects of aerobic or aerobic and resistance training on cardiorespiratory and skeletal muscle function in heart failure: a randomized controlled pilot trial. AB - OBJECTIVE: To examined the effects of different training modalities on exercise capacity (Vo( 2peak)), systolic function, muscular strength and endurance and quality of life in heart failure patients. DESIGN: Randomized controlled trial. SETTING: Cardiac rehabilitation centre in Canada. SUBJECTS: Forty-two individuals with heart failure (62 +/- 12 years; New York Heart Association (NYHA) classes I III). INTERVENTIONS: Aerobic training (n = 14), combined aerobic and resistance training (n = 15) or usual care (n = 13) three times per week for 12 weeks. MAIN MEASURES: (1) Vo( 2peak) measured by symptom-limited graded exercise test on cycle ergometer; (2) systolic function assessed by two-dimensional echocardiography; (3) muscular strength and muscular endurance measured by one repetition maximum procedure; and (4) quality of life assessed by questionnaires. RESULTS: In the intention-to-treat analysis, neither aerobic nor combined aerobic and resistance training significantly improved Vo(2peak), systolic function or quality of life compared with usual care. However, combined aerobic and resistance training significantly improved upper extremity strength (40.7 (14.0) 48.5 (16.0) kg, P<0.05) and muscular endurance (5.7 (2.7)-11.6 (7.6) reps, P<0.05) compared with aerobic training or usual care. In compliant participants (exercise adherence 80%), Vo(2peak) increased in the aerobic group (16.9 (6.0) 19.0 (6.8), P= 0.026) and tended to increase in the combined training group (15.9 (5.0)-17.6 (5.6), P= 0.058) compared with usual care. Quality of life was improved in the aerobic group only. CONCLUSIONS: Both aerobic and combined aerobic and resistance training are effective interventions to improve Vo(2peak) in compliant heart failure patients. Combined training may be more effective in improving muscle strength and endurance. PMID- 19218297 TI - A proof of concept study for the integration of robot therapy with physiotherapy in the treatment of stroke patients. AB - OBJECTIVE: To carry out a proof of concept study for integrating robot therapy with physiotherapy in the treatment of stroke patients. DESIGN: A simple and 'gentle' paradigm of robot-patient interaction was designed in order to foster the re-emergence of smooth, active control patterns in coordinated shoulder/elbow reaching movements. A haptic robot was programmed according to a strategy of minimal, progressively reduced assistance, with a double representation of targets: (i) visual (circles on a screen) and (ii) haptic (robot-generated force fields). The protocol included trials with and without vision, in order to emphasize the role of proprioceptive feedback. The training paradigm included 10 sessions and more than 5000 movements. SUBJECTS: Ten chronic, hemiparetic subjects; four controls provided reference values for the performance measurements. OUTCOME MEASURES: Four performance indicators (derived from the analysis of the reaching trajectories); clinical/functional measures (Fugl-Meyer and Ashworth scales). RESULTS: After robot therapy reaching movements became faster and smoother. The performance in the no-vision trials was at least as good as in the vision trials. The Fugl-Meyer arm scores also increased significantly and remained approximately constant at follow-up; the Ashworth scores did not change. CONCLUSION: In spite of its simplicity, a limited number of ;gentle' robot therapy sessions appear to be beneficial, even for severely impaired patients, although no firm conclusion can be drawn at this point. However, the study provides support material for the careful design of controlled clinical trials and for a better integration with physiotherapy. PMID- 19218298 TI - Comparison of isokinetic exercise versus standard exercise training in patients with chronic low back pain: a randomized controlled study. AB - OBJECTIVE: To compare the effectiveness of an isokinetic exercise programme and a standard exercise programme in patients with chronic low back pain in terms of pain, mobility, disability, psychological status and muscle strength. DESIGN: A randomized controlled trial. SETTING: An outpatient rehabilitation clinic. SUBJECTS: A total of 40 patients with low back pain were included in the study. INTERVENTIONS: Patients with low back pain were randomly allocated into group 1 (n=20, isokinetic exercises) and group 2 (n=20, standard exercise). MAIN MEASURES: Outcome measures included a visual analogue scale (VAS) for pain, fingertip-to-floor test for spinal mobility, Modified Oswestry Low Back Disability Questionnaire (MOLBDQ), Beck Depression Inventory and isokinetic muscle testing. RESULTS: The isokinetic and standard exercise groups demonstrated significant improvement in the VAS, fingertip-to-floor test, MOLBDQ, Beck Depression Inventory scores, and muscle strength compared with the baseline that persisted until the end of the first month (P<0.05). Comparison of both exercise groups in terms of these parameters obtained at the end of the treatment and at the first month after treatment showed no significant difference (P>0.05). CONCLUSION: Isokinetic and standard exercise programmes have an equal effect in the treatment of low back pain, with no statistically significant difference found between the two programmes. The standard exercise programme was easily performed and had a low cost, making it the preferred option for exercise. PMID- 19218299 TI - The effect of walking mobility on the measurement of physical activity using accelerometry in multiple sclerosis. AB - OBJECTIVE: To examine whether accelerometry provides a measure of physical activity, walking ability or both in a sample of individuals with multiple sclerosis. The secondary purpose was to examine the validity of physical activity measures in people with multiple sclerosis who have ambulatory impairments. PARTICIPANTS: Forty-two individuals with multiple sclerosis without ambulatory impairment (Expanded Disability Status Scale (EDSS) score < or = 4.5) and 32 individuals with multiple sclerosis with ambulatory impairment (EDSS > or = 5.0). METHOD: Participants completed the Multiple Sclerosis Walking Scale-12 and Performance Scales, wore an accelerometer for seven days, and completed the Godin Leisure-Time Exercise Questionnaire and short-form of the International Physical Activity Questionnaire. RESULTS: There were significant differences between groups on scores from the five measures. There were large correlations between the accelerometer counts with scores from both the self-report measures of physical activity and the self-report measures of walking mobility in the overall sample. There were large correlations between scores from both measures of physical activity in the overall sample and the subsample with ambulatory impairment. CONCLUSION: Our data suggest that accelerometers are measuring both physical activity and walking mobility in individuals with multiple sclerosis, whereas self-report measures are measuring physical activity in individuals with multiple sclerosis, including those with ambulatory impairment. PMID- 19218300 TI - Predicting accidental falls in people with multiple sclerosis -- a longitudinal study. AB - OBJECTIVE: To investigate accidental falls and near fall incidents in people with multiple sclerosis with respect to clinical variables and the predictive values of four tests. DESIGN: A longitudinal, multi-centred cohort study with prospectively collected falls. PROCEDURES: Self-reported incidents during the three months following a standardized test procedure. SUBJECTS: Seventy-six people with multiple sclerosis and an Expanded Disability Status Scale score between 3.5 and 6.0. MAIN OUTCOME MEASURES: Berg Balance Scale, Timed Up and Go cognitive, Four Square Step Test (FSST) and 12-item Multiple Sclerosis Walking Scale. RESULTS: Forty-eight people (63%) registered 270 falls. Most falls occurred indoors during activities of daily life. We found a correlation of r(s)=0.57 between near falls and falls, and of r(s) = 0.82 between registered and retrospectively recalled falls. Fallers and non-fallers differed significantly regarding Expanded Disability Status Score (odds ratio (OR) 1.99, 95% confidence interval (CI) 1.22; 3.40), spasticity (OR 1.14, CI 1.02; 1.31), proprioception (OR 2.50, CI 1.36; 5.12) and use of walking aids (OR 2.27, CI 1.23; 4.37). Reported use of walking aids both indoors and outdoors increased the odds of falling fivefold while disturbed proprioception increased the odds 2.5-15.6 times depending on severity. The odds of falling were doubled for each degree of increased Expanded Disability Status Score and more than doubled for each degree of increased spasticity. The Berg Balance Scale, use of walking aids and Timed Up and Go cognitive best identified fallers (73-94%) and proprioception, Expanded Disability Status Score, 12-item Multiple Sclerosis Walking Scale and Four Square Step Test best identified non-fallers (75-93%). CONCLUSIONS: In clinical practice, looking at the use of walking aids, investigating proprioception and spasticity, rating Expanded Disability Status Score and using Berg Balance Scale or Timed Up and Go cognitive all contribute when identifying fallers. PMID- 19218301 TI - 'Listen to me, tell me': a qualitative study of partnership in care for people with non-specific chronic low back pain. AB - OBJECTIVE: To investigate and summarize participant experience of exercise programmes for non-specific chronic low back pain and the effects of these experiences on exercise participation and engagement. SUBJECTS: Three groups of six adults (>18 years) who speak, read and understand English and who had participated in an exercise programme for non-specific chronic low back pain. METHOD: Qualitative research methods were used and three focus groups were conducted by an experienced facilitator. Each group was guided with a set of pre determined questions and participants were encouraged to give personal opinions. Transcribed data were read independently by two researchers and analysed thematically according to Grounded Theory. RESULTS: Preference for partnership in care emerged as a significant theme from all focus group transcripts. The following subthemes emerged: (1) engagement with the health care process; (2) 'listen to me; I know my own body'; (3) 'tell me: explain it to me'. People with non-specific chronic low back pain want an active role in their rehabilitation. They expressed anger and frustration at not being listened to, not being provided with adequate explanations and education and not being given credit for knowing their own bodies. Tension existed between patients' wanting a genuine voice in the partnership and them wanting a care-provider to give explicit diagnosis and best management instruction. CONCLUSION: A gap exists between care-seeker experiences of, and preferences for, exercise programmes for back pain. Care seekers consider that care-providers should adopt a willingness to listen and consider care-seeker's experiences when designing exercise programmes. PMID- 19218302 TI - Depression and anxiety symptoms after lower limb amputation: the rise and fall. AB - OBJECTIVE: To examine the time course of anxiety and depressive symptoms over a three year period after amputation. DESIGN AND SETTINGS: A prospective study in inpatients admitted to a rehabilitation ward after lower limb amputation. SUBJECTS: Successive admissions over a one-year period of whom 68 were alive at follow-up, 2-3 years later. INTERVENTIONS: Nil. MAIN MEASURES: Hospital Anxiety and Depression Scale (HADS) on admission and discharge from inpatient rehabilitation and at a 2.7(SD=0.4) year mean follow-up period with correlation to demographic and patient features. RESULTS: Of the 68 responding patients, 12 (17.6%) and 13 (19.1%) had symptoms of depression and anxiety respectively. This compared to an original incidence of 16 (23.5%) for both on admission and 2 (2.9%) on discharge. This rise in incidence from time of discharge was highly significant for both depression (P<0.001) and anxiety (P<0.001). Depression at follow-up was correlated to depressive symptoms at admission (P=0.03) and to having other significant comorbidities (P=0.02). Anxiety symptoms were commoner in younger patients (P=0.03). There was no association with age, gender, living in isolation, vascular cause for amputation, wearing a limb prosthesis or length of original inpatient stay. CONCLUSIONS: Depression and anxiety are common after lower limb amputation but resolve during inpatient rehabilitation. The incidence then rises again after discharge. PMID- 19218303 TI - Skills training: an important component of powered wheelchair use. PMID- 19218304 TI - A risk score to predict disease-free survival in patients not achieving a pathological complete remission after preoperative chemotherapy for breast cancer. AB - BACKGROUND: We aimed to predict disease-free survival (DFS) in patients who failed to achieve a pathologic complete remission (pCR) after preoperative chemotherapy (PC). PATIENTS AND METHODS: Data from 577 patients treated with PC and operated at the European Institute of Oncology (EIO) were used to develop a nomogram using Cox proportional hazards regression model based on both categorical (pT, positive nodes, human epidermal growth factor receptor 2 (HER2) status, vascular invasion) and continuous histological variables (estrogen receptors and Ki-67 expression) at surgery. The nomogram was tested on a second patient cohort (343 patients) treated in other institutions and subsequently operated at the EIO. RESULTS: The nomogram for DFS based on both categorical and continuous variables had good discrimination in the training and the validation sets (concordance indices 0.73, 0.67). CONCLUSION: The use of a nomogram based on the degree of selected histopathological variables can predict DFS and might help in the adjuvant therapeutic algorithm design. PMID- 19218305 TI - Pemetrexed in combination with oxaliplatin as a first-line therapy for advanced gastric cancer: a multi-institutional phase II study. AB - BACKGROUND: This clinical trial assessed the efficacy of pemetrexed combined with oxaliplatin (PEMOX) in patients with advanced gastric cancer (AGC). PATIENTS AND METHODS: Forty-four patients with untreated AGC were enrolled to evaluate response rate (RR). Patients received pemetrexed (500 mg/m(2)) with vitamin supplementation and oxaliplatin (120 mg/m(2)) every 21 days for six cycles or until disease progression occurred. RESULTS: Median age was 62 years (range 26 76). The majority of patients (93%) had metastatic disease. Sixteen of the 44 patients achieved confirmed response [RR 36%; 95% confidence interval (CI) 22% to 52%]; four complete responses and 12 partial responses (complete and partial responses according to the RECIST guidelines are the confirmed-responses observed in the study population). Median time to tumor progression (TTP) was 6.2 months (95% CI 4.3-7.5) and median survival was 10.8 months (95% CI 7.7-17.2). A total of 220 cycles were administered, with a median of six cycles. Most common grade 3/4 toxic effects were neutropenia in 41% of patients (19% of cycles) and thrombocytopenia in 11% of patients (4% of cycles). Treatment delays or dose reductions for toxicity occurred in 10% and 5% of cycles, respectively. CONCLUSIONS: PEMOX is active and well tolerated in AGC. RR, TTP, and survival were comparable to those achieved in studies using different 5-fluorouracil (5 FU)-oxaliplatin combinations, without the inconvenience of prolonged 5-FU schedules. PMID- 19218306 TI - Effects of exemestane and tamoxifen on bone health within the Tamoxifen Exemestane Adjuvant Multicentre (TEAM) trial: results of a German, 12-month, prospective, randomised substudy. AB - BACKGROUND: Adjuvant treatment of hormone receptor-positive breast cancer in postmenopausal women with aromatase inhibitors may be associated with increased bone loss. PATIENTS AND METHODS: Two hundred patients were randomised to receive exemestane or tamoxifen as adjuvant treatment of hormone receptor-positive breast cancer. Bone mineral density (BMD) was assessed by dual-energy X-ray absorptiometry at baseline and after 6 and 12 months treatment. RESULTS: One hundred and sixty-one patients were assessable. Tamoxifen treatment resulted in a 0.5% increase from baseline in BMD at the spine, which was maintained at 12 months. Exemestane-treated patients experienced a 2.6% decrease from baseline in BMD at the spine at 6 months and a further 0.2% decrease at 12 months. There were significant differences in the changes in BMD between tamoxifen and exemestane at 6 and 12 months (P = 0.0026 and P = 0.0008, respectively). The mean changes in BMD from baseline at the total hip were also significantly different between exemestane and tamoxifen at 6 and 12 months (P = 0.0009 and P = 0.04, respectively). There was no difference between tamoxifen and exemestane in mean changes in BMD from baseline at the femoral neck. CONCLUSIONS: Exemestane treatment resulted in an increase in bone loss at 6 months; bone loss stabilised after 6- to 12-month treatment. PMID- 19218307 TI - Preoperative weekly cisplatin-epirubicin-paclitaxel with G-CSF support in triple negative large operable breast cancer. AB - BACKGROUND: Findings from our previously published phase II study showed a high pathologic complete remission (pCR) rate in patients with triple-negative large operable breast cancer after the administration of eight cisplatin-epirubicin paclitaxel (PET) weekly cycles. The safety and efficacy data of the initial population were updated, with inclusion of additional experience with the same therapy. METHODS: Patients with triple-negative large operable breast cancer (T2 T3 N0-1; T > 3 cm) received eight preoperative weekly cycles of cisplatin 30 mg/m2, epirubicin 50 mg/m2, paclitaxel (Taxol) 120 mg/m2, with granulocyte colony stimulating factor (5 microg/kg days 3-5) support. RESULTS: Overall 74 consecutive patients (T2/T3 = 35/39; N0/N+ = 26/48) were treated, from May 1999 to May 2008. At pathological assessment, 46 women (62%; 95% confidence interval 50-73) showed pCR in both breast and axilla. At a 41-month median follow-up (range 3-119), 13 events (nine distant metastases) had occurred, 5-year projected disease-free survival (DFS) and distant disease-free survival being 76% and 84%, respectively. Five-year DFS was 90% and 56% in pCRs and non-pCRs, respectively. Severe neutropenia and anemia occurred in 23 (31%) and eight (10.8%) patients, respectively. Severe non-hematological toxicity was recorded in <20% of patients. Peripheral neuropathy was quite frequent but never severe. CONCLUSIONS: Eight weekly PET cycles are a highly effective primary treatment in women with triple negative large operable breast cancer. This approach results in a very promising long-term DFS in this poor prognosis population. This triplet regimen is worthy of evaluation in phase III trials. PMID- 19218308 TI - Supporting nurse decision making in primary care: exploring use of and attitude to decision tools. AB - Nurses are increasingly working more autonomously in extended roles, yet we know little about the nature of the decisions they make. Decisions vary in terms of complexity, ambiguity and presentation, and the nature of the decision task impacts on the process of decision making, such as the likelihood of using a decision tool. Thus, knowledge about the nature of nursing decisions is essential for development of effective decision tools. This article presents an analysis of 410 nurse-patient consultations and interviews with 76 primary care nurses, and explores the nature of the decisions that primary care nurses make and the impact of that on their use of and attitudes towards decision tools. PMID- 19218309 TI - Mobile phone text messaging to promote healthy behaviors and weight loss maintenance: a feasibility study. AB - There is a need to investigate newer strategies pertaining to the maintenance of healthy behaviors and weight. We investigated the feasibility of mobile phone text messaging to enable ongoing communication with African-American women participating in a weight management program. Ninety-five African-American women participated in this pilot study and received regularly scheduled text messages. Forty-two of these women chose to create 165 personal text messages that included tips on healthy eating and physical activity, as well as reminders to drink water and expressions of encouragement. A commercially available client-based application transmitted these personal messages and general health messages at least three times per week. The software transmitted over 4500 text messages during the first 4 months with 114 returned as undeliverable. Participants expressed generally positive attitudes toward incoming text messages, with only one participant declining to continue after enrollment. This study demonstrated early feasibility and acceptability of text messaging as a method for promoting healthy behaviors for weight maintenance. PMID- 19218310 TI - Usability and acceptability of a website that provides tailored advice on falls prevention activities for older people. AB - This article presents the usability and acceptability of a website that provides older people with tailored advice to help motivate them to undertake physical activities that prevent falls. Views on the website from interviews with 16 older people and 26 sheltered housing wardens were analysed thematically. The website was well received with only one usability difficulty with the action plan calendar. The older people selected balance training activities out of interest or enjoyment, and appeared to carefully add them into their current routine. The wardens were motivated to promote the website to their residents, particularly those who owned a computer, had balance problems, or were physically active. However, the participants noted that currently a minority of older people use the Internet. Also, some older people underestimated how much activity was enough to improve balance, and others perceived themselves as too old for the activities. PMID- 19218311 TI - A means to an end: a web-based client management system in palliative care. AB - Home-based palliative care (hospice) services require comprehensive and fully integrated information systems to develop and manage the various aspects of their business, incorporating client data and management information. These systems assist in maintaining the quality of client care as well as improved management efficiencies. This article reports on a large not-for-profit home-based palliative care service in Australia, which embarked on a project to develop an electronic data management system specifically designed to meet the needs of the palliative care sector. This web-based client information management system represents a joint venture between the organization and a commercial company and has been a very successful project. PMID- 19218312 TI - Prescription for nursing informatics in pre-registration nurse education. AB - Nurses need to be able to use information and communications technology not only to support their own practice, but also to help their patients make best use of it. This article argues that nurses are not currently adequately prepared to work with information and technology through their pre-registration education. Reflecting the lack of nursing informatics expertise, it is recommended that all pre-registration nursing programmes should have access to a nursing informatics specialist. A prescription to meet the informatics needs of the newly qualified nurse is proposed. This places the areas that need to be included in pre registration education into broad groups that both articulate the competencies that nurses need to develop, and indicate why they are needed, rather than providing context-free checklists of skills. This is presented as a binary scatter chart with two axes, skill to knowledge and technology to information. PMID- 19218313 TI - The obstetrician and gynaecologist and the European Computer Driving Licence. AB - The European Computer Driving Licence (ECDL) has been adopted as the reference standard for all NHS staff for basic IT skills. The objective of the study was to determine awareness of the ECDL amongst members of the Royal College of Obstetricians and Gynaecologists. An anonymous postal survey was conducted with members and fellows of the College in three UK regions. A total of 720 questionnaires were posted, and the response rate was 47.9 per cent. A fifth of all respondents were not aware of the ECDL at all, and only 10 per cent were trained in the ECDL. However, more than third had some other formal training in IT skills. Half the respondents suggested that the questionnaire had prompted them to train for the ECDL. Much needs to be done to increase awareness of the ECDL and offer incentives for its takeup. PMID- 19218314 TI - The putative RNA-processing protein, THO2, is a microtubule-associated protein in tobacco. AB - THO2 is a component of the THO-TREX (transcription and export factor) complex that participates in mRNA metabolism and export from the nucleus in yeast and animal cells. Here we report that tobacco putative THO2-related protein (NtTHO2) is a microtubule-associated protein, which directly binds to microtubules in vitro and co-localizes with cortical microtubules in vivo. We purified endogenous NtTHO2 by cycles of microtubule polymerization-depolymerization from crude extracts of tobacco BY-2 miniprotoplasts. Purified NtTHO2 sedimented with microtubules in vitro. Immunofluorescence revealed that NtTHO2 was present in both the nucleus and cytoplasm. In interphase, cytoplasmic NtTHO2 was localized along cortical microtubules. In the mitotic phase, NtTHO2 was localized to the mitotic spindle but not to either the preprophase band or the phragmoplast. In mature cells of seedling roots, and in BY-2 cells in which proliferation was stopped by removing 2,4-D, NtTHO2 staining was confined mainly to the nucleolus. These results suggest that NtTHO2 is a multifunctional protein that participates in mRNA metabolism, and also functions within the cortical microtubules and mitotic spindle. PMID- 19218315 TI - Transcriptome analysis of a spontaneous mutant in sweet orange [Citrus sinensis (L.) Osbeck] during fruit development. AB - Bud mutations often arise in citrus. The selection of mutants is one of the most important breeding channels in citrus. However, the molecular basis of bud mutation has rarely been studied. To identify differentially expressed genes in a spontaneous sweet orange [C. sinensis (L.) Osbeck] bud mutation which causes lycopene accumulation, low citric acid, and high sucrose in fruit, suppression subtractive hybridization and microarray analysis were performed to decipher this bud mutation during fruit development. After sequencing of the differentially expressed clones, a total of 267 non-redundant transcripts were obtained and 182 (68.2%) of them shared homology (E-value < or = 1x10(-10)) with known gene products. Few genes were constitutively up- or down-regulated (fold change > or = 2) in the bud mutation during fruit development. Self-organizing tree algorithm analysis results showed that 95.1% of the differentially expressed genes were extensively coordinated with the initiation of lycopene accumulation. Metabolic process, cellular process, establishment of localization, response to stimulus, and biological regulation-related transcripts were among the most regulated genes. These genes were involved in many biological processes such as organic acid metabolism, lipid metabolism, transport, and pyruvate metabolism, etc. Moreover, 13 genes which were differentially regulated at 170 d after flowering shared homology with previously described signal transduction or transcription factors. The information generated in this study provides new clues to aid in the understanding of bud mutation in citrus. PMID- 19218316 TI - Two distinct intracellular Ca2+-release components act in opposite ways in the regulation of the auxin-dependent MIA biosynthesis in Catharanthus roseus cells. AB - Calcium-mediated signalling is ubiquitous in both animals and plants. Changes in cytoplasmic free Ca(2+) concentration couple diverse arrays of stimuli to their specific responses, the specificity of the stimulus being determined by integrated actions between multiple Ca(2+) mobilization pathways. In this work, a pharmacological approach is reported, aimed at deciphering the role of calcium as a second messenger in the transduction pathway leading to the inhibitory effect of 2,4-dichlorophenoxyacetic acid (2,4-D), in regulating monoterpene indole alkaloid (MIA) biosynthesis in Catharanthus roseus cells. It is demonstrated here that auxin-dependent MIA biosynthesis is differentially regulated by two distinct calcium release components from internal stores in C. roseus showing pharmacological profiles similar to those displayed by animal RyR and IP3 channels. MIA biosynthesis is stimulated by caffeine (Ca(2+)-release activator through RyR channels) and by heparin and TMB8 (Ca(2+)-release inhibitors of IP3 channels) whereas MIA biosynthesis is inhibited by mastoparan (Ca(2+)-release activator of IP3 channels) and by ruthenium red and DHBP (Ca(2+)-release inhibitors of RyR channels). Furthermore, calcium, as 2,4-D, acts on MIA biosynthesis by regulating the monoterpene moiety of the MIA biosynthesis pathway since calcium channel modulators preferentially modulate g10h expression, the gene encoding the enzyme of the secoiridoid monoterpene pathway, that is the major target of 2,4-D action. In addition, the simultaneous use of caffeine (an activator of RyR channel in animals) and TMB8 (an inhibitor of the IP3 channel) in 2,4-D treated cells triggers a synergistic effect on MIA accumulation. This finding suggests an opposite and co-ordinated action of multiple Ca(2+)-release pathways in 2,4-D signal transduction, adding a new level of complexity to calcium signalling in plants and questioning the existence of RyR and IP3 channels in plants. PMID- 19218317 TI - Potential of Jatropha curcas as a source of renewable oil and animal feed. AB - Jatropha curcas (L.) is a perennial plant of the spurge family (Euphorbiaceae). Recently, it has received much attention as a potential source of vegetable oil as a replacement for petroleum, and, in particular, the production of biodiesel. Despite the interest that is being shown in the large-scale cultivation of J. curcas, genetic resources remain poorly characterized and conserved and there has been very little plant breeding for improved traits. At present, the varieties being used to establish plantations in Africa and Asia are inedible. The meal obtained after the extraction of oil cannot, therefore, be used as a source of animal feed. Naturally existing edible varieties are, however, known to occur in Mexico. The toxic components of J. curcas seeds, the potential for plant breeding to generate improved varieties, and the suitability of J. curcas oil as a feedstock for biodiesel production are discussed. PMID- 19218318 TI - Defining polycystic ovary syndrome. PMID- 19218319 TI - Beyond the impact factor. PMID- 19218320 TI - Increasing CD4 count with interleukin-2 does not benefit people with HIV. PMID- 19218321 TI - NHS needs to prepare for era of non-invasive fetal DNA tests. PMID- 19218322 TI - Discovery of new cause for fractures among children who were adopted has "painful" lessons for doctors. PMID- 19218323 TI - Comparison of different strategies in prenatal screening for Down's syndrome: cost effectiveness analysis of computer simulation. AB - OBJECTIVES: To assess and compare the cost effectiveness of three different strategies for prenatal screening for Down's syndrome (integrated test, sequential screening, and contingent screenings) and to determine the most useful cut-off values for risk. DESIGN: Computer simulations to study integrated, sequential, and contingent screening strategies with various cut-offs leading to 19 potential screening algorithms. DATA SOURCES: The computer simulation was populated with data from the Serum Urine and Ultrasound Screening Study (SURUSS), real unit costs for healthcare interventions, and a population of 110 948 pregnancies from the province of Quebec for the year 2001. MAIN OUTCOME MEASURES: Cost effectiveness ratios, incremental cost effectiveness ratios, and screening options' outcomes. RESULTS: The contingent screening strategy dominated all other screening options: it had the best cost effectiveness ratio ($C26,833 per case of Down's syndrome) with fewer procedure related euploid miscarriages and unnecessary terminations (respectively, 6 and 16 per 100,000 pregnancies). It also outperformed serum screening at the second trimester. In terms of the incremental cost effectiveness ratio, contingent screening was still dominant: compared with screening based on maternal age alone, the savings were $C30,963 per additional birth with Down's syndrome averted. Contingent screening was the only screening strategy that offered early reassurance to the majority of women (77.81%) in first trimester and minimised costs by limiting retesting during the second trimester (21.05%). For the contingent and sequential screening strategies, the choice of cut-off value for risk in the first trimester test significantly affected the cost effectiveness ratios (respectively, from $C26,833 to $C37,260 and from $C35,215 to $C45,314 per case of Down's syndrome), the number of procedure related euploid miscarriages (from 6 to 46 and from 6 to 45 per 100,000 pregnancies), and the number of unnecessary terminations (from 16 to 26 and from 16 to 25 per 100,000 pregnancies). CONCLUSIONS: Contingent screening, with a first trimester cut-off value for high risk of 1 in 9, is the preferred option for prenatal screening of women for pregnancies affected by Down's syndrome. PMID- 19218324 TI - Prenatal screening for Down's syndrome. PMID- 19218325 TI - Late onset hypogonadism. PMID- 19218326 TI - UK government rejects advice from drugs adviser to downgrade ecstasy. PMID- 19218327 TI - Home secretary accused of bullying drugs adviser over comments about ecstasy. PMID- 19218328 TI - World Bank provides $2.4bn to pay mothers to take their babies to health clinics. PMID- 19218329 TI - GMC consults on dropping vexatious complaints at an early stage. PMID- 19218330 TI - Removal of red light minimizes methylene blue-stimulated DNA damage in oesophageal cells: implications for chromoendoscopy. AB - Barrett's oesophagus (BO) carries an increased risk of progression to oesophageal adenocarcinoma. Chromoendoscopy with methylene blue (MB) can be used to facilitate identification of BO and target areas for biopsy. If photoexcited, MB can generate reactive oxygen species and genotoxic photodegradation products leading to DNA damage. We have previously demonstrated that levels of DNA damage are increased in BO following MB chromoendoscopy. The aim of this study was to investigate whether DNA damage, as measured by the comet assay, can be minimized during chromoendoscopy by varying MB concentration and light wavelength using an in vitro model. OE33 cells were treated with MB (0.015-15 mM) and exposed to white light (WL). Cells were also illuminated with WL fractions (580-700, 480 580, 350-480, <575, <610 and <688 nm) in the presence of MB. At clinically relevant concentrations, WL illumination of MB (15 mM) caused significant DNA damage in vitro (P < 0.001). Illumination of MB with red light (580-700 nm) also stimulated high levels of DNA damage in OE33 cells (P < 0.001). This effect was not observed with green or blue light. Filtering WL to remove red light wavelengths (>575 nm) reduced DNA damage and apoptosis to control levels in MB treated cells. In addition, reducing the concentration of MB 10-fold markedly reduced the DNA-damaging effect of MB in vitro. The results show that photoactivation of MB by red light is responsible for the majority of DNA damage. Simple modifications to MB chromoendoscopy, such as filtering out red light from endoscopic WL or reducing MB concentration, are likely to limit DNA damage induced by the procedure. PMID- 19218331 TI - Bisphenol A disrupts Notch signaling by inhibiting gamma-secretase activity and causes eye dysplasia of Xenopus laevis. AB - Bisphenol A (BPA) is being recognized as an endocrine-disrupting chemical (EDC). Recently, several reports indicated that BPA affects the central nervous system (CNS) during embryonic development. However, the molecular mechanism of BPA in the CNS is not well known. Here, we show that BPA affected Notch signaling by inhibiting the activity of the Notch intracellular domain (NICD) cleavage-related enzyme, gamma-secretase (gamma-secretase), at the neurula stage of the Xenopus laevis. BPA caused various morphologic aberrations including scoliosis, eye dysplasia, and loss of pigments in the X. laevis tadpole. These abnormalities were seen whenever BPA was used at the neurula stage. In addition, the expression levels of several marker mRNAs at the neurula stage were investigated by RT-PCR, and we found that the mRNAs expression of ectodermal marker, Pax6, CNS marker, Sox2, and neural crest marker, FoxD3, were decreased by treatment with BPA. These genes contribute to the neural differentiation at the neurula stage, and also the downstream factors of Notch signaling. Injection of NICD but not a Notch ligand, delta 1, rescued the abnormalities caused by BPA. We subsequently assayed the inhibition of the activities of NICD cleavage-related enzymes, tumor necrosis factor alpha converting enzyme, and gamma-secretase, by BPA and found that BPA inhibited the gamma-secretase activity. Furthermore, we expressed presenilin, a main component of gamma-secretase, in Escherichia coli and found the direct binding of BPA with presenilin. These results suggest that BPA affected the neural differentiation by inhibiting gamma-secretase activity, leading to neurodevelopmental abnormalities. PMID- 19218332 TI - Estimating inequalities in ownership of insecticide treated nets: does the choice of socio-economic status measure matter? AB - Research on the impact of socio-economic status (SES) on access to health care services and on health status is important for allocating resources and designing pro-poor policies. Socio-economic differences are increasingly assessed using asset indices as proxy measures for SES. For example, several studies use asset indices to estimate inequities in ownership and use of insecticide treated nets as a way of monitoring progress towards meeting the Abuja targets. The validity of different SES measures has only been tested in a limited number of settings, however, and there is little information on how choice of welfare measure influences study findings, conclusions and policy recommendations. In this paper, we demonstrate that household SES classification can depend on the SES measure selected. Using data from a household survey in coastal Kenya (n = 285 rural and 467 urban households), we first classify households into SES quintiles using both expenditure and asset data. Household SES classification is found to differ when separate rural and urban asset indices, or a combined asset index, are used. We then use data on bednet ownership to compare inequalities in ownership within each setting by the SES measure selected. Results show a weak correlation between asset index and monthly expenditure in both settings: wider inequalities in bednet ownership are observed in the rural sample when expenditure is used as the SES measure [Concentration Index (CI) = 0.1024 expenditure quintiles; 0.005 asset quintiles]; the opposite is observed in the urban sample (CI = 0.0518 expenditure quintiles; 0.126 asset quintiles). We conclude that the choice of SES measure does matter. Given the practical advantages of asset approaches, we recommend continued refinement of these approaches. In the meantime, careful selection of SES measure is required for every study, depending on the health policy issue of interest, the research context and, inevitably, pragmatic considerations. PMID- 19218333 TI - Impact of exercise training on myostatin expression in the myocardium and skeletal muscle in a chronic heart failure model. AB - AIMS: In late-stage chronic heart failure (CHF), elevated cytokines and cachexia are often observed. Several studies have shown that exercise training exerts beneficial effects on skeletal muscle in this setting. Furthermore, it has been shown that the expression of myostatin, a key regulator of skeletal muscle mass, is increased in a variety of cachectic states. This study aimed to investigate the expression of myostatin in CHF, the influence of exercise training on myostatin levels, and regulation of myostatin by tumour necrosis factor-alpha (TNF-alpha). METHODS AND RESULTS: In an animal model of CHF (LAD-ligation model), protein expression of myostatin was elevated 2.4-fold in the skeletal muscle and more than four-times in the myocardium, compared with control (Co). Exercise training on a treadmill over 4 weeks led to a significant reduction in myostatin protein expression in the skeletal muscle and the myocardium of CHF animals, with values returning to baseline levels. In differentiated C2C12 cells, TNF-alpha induced the expression of myostatin through a p38MAPK-dependent pathway involving nuclear factor kappa-B (NF-kappaB). The increased TNF-alpha mRNA levels in the skeletal muscle of CHF animals correlated significantly with myostatin expression. CONCLUSION: These alterations in myostatin expression in the skeletal and heart muscle following exercise training could help to explain the beneficial anti-catabolic effects of exercise training in CHF. PMID- 19218334 TI - Improving data quality control in quality improvement projects. AB - BACKGROUND: The results of many quality improvement (QI) projects are gaining wide-spread attention. Policy-makers, hospital leaders and clinicians make important decisions based on the assumption that QI project results are accurate. However, compared with clinical research, QI projects are typically conducted with substantially fewer resources, potentially impacting data quality. Our objective was to provide a primer on basic data quality control methods appropriate for QI efforts. METHODS: Data quality control methods should be applied throughout all phases of a QI project. In the design phase, project aims should guide data collection decisions, emphasizing quality (rather than quantity) of data and considering resource limitations. In the data collection phase, standardized data collection forms, comprehensive staff training and a well-designed database can help maximize the quality of the data. Clearly defined data elements, quality assurance reviews of both collection and entry and system based controls reduce the likelihood of error. In the data management phase, missing data should be quickly identified and corrected with system-based controls to minimize the missing data. Finally, in the data analysis phase, appropriate statistical methods and sensitivity analysis aid in managing and understanding the effects of missing data and outliers, in addressing potential confounders and in conveying the precision of results. CONCLUSION: Data quality control is essential to ensure the integrity of results from QI projects. Feasible methods are available and important to help ensure that stakeholder's decisions are based on accurate data. PMID- 19218335 TI - Adolescents perceived effectiveness of the proposed European graphic tobacco warning labels. AB - BACKGROUND: Graphical tobacco product labelling is a prominent source of health information and has an important position among tobacco control initiatives. However, little is known about its effectiveness among adolescents. With this above in mind, we aimed to research into how adolescents perceive the proposed EU graphic tobacco product warning labels as an effective means of preventing smoking initiation in comparison to the current EU text-only warning labels. METHODS: Five hundred seventy four adolescents (13-18, 54% male) from Greece were privately interviewed, with the use of a digital questionnaire and randomly shown seven existing EU text-only and proposed EU graphic warning labels. Non-smoking respondents were asked to compare and rate the warnings effectiveness in regard to preventing them from smoking on a 1-5 Likert type scale. RESULTS: Irrespective of the warning category shown, on all occasions, non-smoking adolescents rated the suggested EU graphic labels as more effective in preventing them from smoking in comparison to the existing EU text-only warnings. Controlling for gender, age, current smoking status and number of cigarettes smoked per month, younger adolescents were found to opt for graphic warnings more often, and also perceive graphic warning labels as a more effective means of preventing them from smoking, in comparison to their elder peers (P < 0.001). CONCLUSIONS: The proposed EU graphic warning labels may play an important role in preventing of smoking initiation during the crucial years of early adolescence when smoking experimentation and early addiction usually take place. PMID- 19218336 TI - Matrix metalloproteinase-12 deficiency worsens relapsing-remitting experimental autoimmune encephalomyelitis in association with cytokine and chemokine dysregulation. AB - The elevation of several members of the matrix metalloproteinase (MMP) family promotes the pathophysiology of both multiple sclerosis and its animal model, experimental autoimmune encephalomyelitis (EAE). Nonetheless, given the multiple activities of MMPs, it remains possible that increased levels of a particular MMP may have beneficial functions during disease progression. We reported previously that MMP-12(-/-) mice of the 129/SvEv strain had a poorer EAE outcome than wild type controls. However, we did not determine further differences in disease profiles between these groups. Using the EAE model in 129/SvEv mice, we report that disease in both wild-type and MMP-12(-/-) mice follows a relapsing-remitting course. Although both mouse groups had similar clinical onsets, subsequent relapses were more severe in MMP-12(-/-) mice; their residual disability at remission was also higher compared with wild-type controls. The worsened relapses and remissions in MMP-12(-/-) mice occurred despite a deficiency of the antigen recall capacity of lymph node-derived cells as well as a reduction in the proportion of macrophages in the spinal cord during the chronic phase of EAE. Significantly, large increases of levels of chemokines and cytokines were found in the spinal cords of MMP-12(-/-) mice during chronic EAE. These results highlight MMP-12 as a beneficial enzyme in EAE and suggest that therapeutic interventions in multiple sclerosis should avoid targeting MMP-12. PMID- 19218337 TI - Spontaneous metastasis of prostate cancer is promoted by excess hyaluronan synthesis and processing. AB - Accumulation of extracellular hyaluronan (HA) and its processing enzyme, the hyaluronidase Hyal1, predicts invasive, metastatic progression of human prostate cancer. To dissect the roles of hyaluronan synthases (HAS) and Hyal1 in tumorigenesis and metastasis, we selected nonmetastatic 22Rv1 prostate tumor cells that overexpress HAS2, HAS3, or Hyal1 individually, and compared these cells with co-transfectants expressing Hyal1 + HAS2 or Hyal1 + HAS3. Cells expressing only HAS were less tumorigenic than vector control transfectants on orthotopic injection into mice. In contrast, cells co-expressing Hyal1 + HAS2 or Hyal1 + HAS3 showed greater than sixfold and twofold increases in tumorigenesis, respectively. Fluorescence and histological quantification revealed spontaneous lymph node metastasis in all Hyal1 transfectant-implanted mice, and node burden increased an additional twofold when Hyal1 and HAS were co-expressed. Cells only expressing HAS were not metastatic. Thus, excess HA synthesis and processing in concert accelerate the acquisition of a metastatic phenotype by prostate tumor cells. Intratumoral vascularity did not correlate with either tumor size or metastatic potential. Analysis of cell cycle progression revealed shortened doubling times of Hyal1-expressing cells. Both adhesion and motility on extracellular matrix were diminished in HA-overproducing cells; however, motility was increased twofold by Hyal1 expression and fourfold to sixfold by Hyal1/HAS co expression, in close agreement with observed metastatic potential. This is the first comprehensive examination of these enzymes in a relevant prostate cancer microenvironment. PMID- 19218339 TI - Identification of potential therapeutic targets in malignant mesothelioma using cell-cycle gene expression analysis. AB - Cell-cycle defects are responsible for cancer onset and growth. We studied the expression profile of 60 genes involved in cell cycle in a series of malignant mesotheliomas (MMs), normal pleural tissues, and MM cell cultures using a quantitative polymerase chain reaction-based, low-density array. Nine genes were significantly deregulated in MMs compared with normal controls. Seven genes were overexpressed in MMs, including the following: CDKN2C, cdc6, cyclin H, cyclin B1, CDC2, FoxM1, and Chk1, whereas Ube1L and cyclin D2 were underexpressed. Chk1 is a principal mediator of cell-cycle checkpoints in response to genotoxic stress. We confirmed the overexpression of Chk1 in an independent set of 87 MMs by immunohistochemistry using tissue microarrays. To determine whether Chk1 down regulation would affect cell-cycle control and cell survival, we transfected either control or Chk1 siRNA into two mesothelioma cell lines and a nontumorigenic (Met5a) cell line. Results showed that Chk1 knockdown increased the apoptotic fraction of MM cells and induced an S phase block in Met5a cells. Furthermore, Chk1 silencing sensitized p53-null MM cells to both an S phase block and apoptosis in the presence of doxorubicin. Our results indicate that cell cycle gene expression analysis by quantitative polymerase chain reaction can identify potential targets for novel therapies. Chk1 knockdown could provide a novel therapeutic approach to arrest cell-cycle progression in MM cells, thus increasing the rate of cell death. PMID- 19218340 TI - Cyclooxygenase-2 is involved in the up-regulation of matrix metalloproteinase-9 in cholangiocarcinoma induced by tumor necrosis factor-alpha. AB - Matrix metalloproteinase-9 (MMP-9) is an important enzyme in tumor invasion and metastasis in malignant tumors, including cholangiocarcinoma (CC). Tumor necrosis factor-alpha (TNF-alpha), a proinflammatory cytokine, was recently reported to induce the up-regulation of MMP-9 in cultured CC cells. We examined whether cyclooxygenase-2 (COX-2) and prostaglandin-E2 (PGE2), another endogenous tumor promoter, are involved in the up-regulation of MMP-9 in CC using CC tissue specimens and a CC cell line, HuCCT-1. MMP-9 and COX-2 were immunohistochemically expressed in 58% and 89% of 110 CC cases, respectively; the expression of MMP-9 and COX-2 was correlated (r = 0.32, P = 0.00072). Using zymography, latent MMP-9 was detectable in all cases and active MMP-9 was detected in 24% of cases of the CC specimens. The TNF-alpha/TNF-receptor 1 (TNF-R1) interaction induced MMP-9 production and activation, as well as COX-2 overexpression and PGE2 production, and increased the migration of CC cells. MMP-9 up-regulation was inhibited by COX inhibitors, antagonists of EP2/4 (receptors of PGE2), and COX-1 and COX-2 siRNAs. Inhibitors of both MMP-9 and MMP-9 siRNA treatment abrogated the increase in the migration of CC cells induced by TNF-alpha. In conclusion, we propose a novel signaling pathway of MMP-9 up-regulation in CC cells such that TNF-alpha induces the activation of COX-2 and PGE2 via TNF-R1 followed by the up-regulation of MMP 9 via the PGE2 (EP2/4) receptor. PMID- 19218341 TI - High-density gene expression analysis of tumor-associated macrophages from mouse mammary tumors. AB - Clinical and experimental evidence indicates that tumor-associated macrophages (TAMs) promote malignant progression. In breast cancer, TAMs enhance tumor angiogenesis, tumor cell invasion, matrix remodeling, and immune suppression against the tumor. In this study, we examined late-stage mammary tumors from a transgenic mouse model of breast cancer. We used flow cytometry under conditions that minimized gene expression changes to isolate a rigorously defined TAM population previously shown to be associated with invasive carcinoma cells. The gene expression signature of this population was compared with a similar population derived from spleens of non-tumor-bearing mice using high-density oligonucleotide arrays. Using stringent selection criteria, transcript abundance of 460 genes was shown to be differentially regulated between the two populations. Bioinformatic analyses of known functions of these genes indicated that formerly ascribed TAM functions, including suppression of immune activation and matrix remodeling, as well as multiple mediators of tumor angiogenesis, were elevated in TAMs. Further bioinformatic analyses confirmed that a pure and valid TAM gene expression signature in mouse tumors could be used to assess expression of TAMs in human breast cancer. The data derived from these more physiologically relevant autochthonous tumors compared with previous studies in tumor xenografts suggest tactics by which TAMs may regulate tumor angiogenesis and thus provide a basis for exploring other transcriptional mediators of TAM trophic functions within the tumor microenvironment. PMID- 19218342 TI - E22Q-mutant Abeta peptide (AbetaDutch) increases vascular but reduces parenchymal Abeta deposition. AB - Patients that have hereditary cerebral hemorrhage with amyloidosis-Dutch type (HCHWA-D) generate both wild-type beta-amyloid (Abetawt) and E22Q-mutant beta amyloid (AbetaDutch). Postmortem analysis of HCHWA-D brains reveals severe cerebral amyloid angiopathy with very little parenchymal amyloid deposition. To investigate amyloidosis in the presence of both Abetawt and AbetaDutch variants, transgenic (tg) APP23 mice were crossed with APPDutch mice. Although single-tg APP23 mice deposited Abetawt with aging, double-tg APP23/APPDutch mice co deposited AbetaDutch (mainly AbetaDutch1-40) and Abetawt at twofold higher total Abeta levels. Vascular Abeta deposits and hemorrhages were twice as high in APP23/APPDutch mice compared with APP23 mice. Surprisingly, parenchymal Abeta deposition was reduced in the double-tg mice compared with the single-tg APP23 mice. Our findings suggest that AbetaDutch1-40 inhibits parenchymal amyloidosis but exacerbates vascular amyloid, hence explaining the compartment-specific distribution of cerebral amyloid in HCHWA-D patients. PMID- 19218343 TI - Roles and mechanisms of human immunodeficiency virus protease inhibitor ritonavir and other anti-human immunodeficiency virus drugs in endothelial dysfunction of porcine pulmonary arteries and human pulmonary artery endothelial cells. AB - The objective of this study was to determine the effects of highly active antiretroviral therapy (HAART) drugs on pulmonary endothelial function. Porcine pulmonary arteries or human pulmonary arterial endothelial cells (HPAECs) were incubated with eight HAART drugs [ritonavir, indinavir, lopinavir, zidovudine (AZT), abacavir, stavudine, didanosine (ddI), and lamivudine] individually or in combination [three HAART drugs (3-plex; indinavir, stavudine, and ddI)] at their clinical plasma concentrations for 24 hours. Endothelium-dependent vasorelaxation in response to bradykinin was reduced significantly by the ritonavir in a concentration-dependent manner. Five other HAART drugs (indinavir, lamivudine, abacavir, AZT, and ddI) and the 3-plex significantly also impaired endothelium dependent vasorelaxation in response to bradykinin. Five HAART drugs (ritonavir, indinavir, lamivudine, abacavir, and AZT) significantly decreased endothelial nitric oxide synthase (eNOS) expression and increased superoxide anion levels in both vessels and HPAECs. Furthermore, both ritonavir and AZT substantially activated ERK2 in HPAECs. Additionally, the antioxidants ginsenoside Rb1 and ginkgolide A effectively reversed HAART drug-induced vasomotor dysfunction and eNOS down-regulation. Inhibition of ERK1/2 also partially blocked ritonavir- and AZT-induced down-regulation of eNOS and vasomotor dysfunction. Thus, HAART drugs significantly impair endothelial functions of porcine pulmonary arteries and HPAECs, which may be mediated by eNOS down-regulation, oxidative stress, and ERK1/2 activation. These findings suggest that HAART drugs may contribute to the high incidence of pulmonary artery hypertension in human immunodeficiency virus infected patients. PMID- 19218344 TI - Identification and characterization of aortic valve mesenchymal progenitor cells with robust osteogenic calcification potential. AB - Advanced valvular lesions often contain ectopic mesenchymal tissues, which may be elaborated by an unidentified multipotent progenitor subpopulation within the valve interstitium. The identity, frequency, and differentiation potential of the putative progenitor subpopulation are unknown. The objectives of this study were to determine whether valve interstitial cells (VICs) contain a subpopulation of multipotent mesenchymal progenitor cells, to measure the frequencies of the mesenchymal progenitors and osteoprogenitors, and to characterize the osteoprogenitor subpopulation because of its potential role in calcific aortic valve disease. The multilineage potential of freshly isolated and subcultured porcine aortic VICs was tested in vitro. Progenitor frequencies and self-renewal capacity were determined by limiting dilution and colony-forming unit assays. VICs were inducible to osteogenic, adipogenic, chondrogenic, and myofibrogenic lineages. Osteogenic differentiation was also observed in situ in sclerotic porcine leaflets. Primary VICs had strikingly high frequencies of mesenchymal progenitors (48.0 +/- 5.7%) and osteoprogenitors (44.1 +/- 12.0%). High frequencies were maintained for up to six population doublings, but decreased after nine population doublings to 28.2 +/- 9.9% and 5.8 +/- 1.3%, for mesenchymal progenitors and osteoprogenitors, respectively. We further identified the putative osteoprogenitor subpopulation as morphologically distinct cells that occur at high frequency, self-renew, and elaborate bone matrix from single cells. These findings demonstrate that the aortic valve is rich in a mesenchyma l progenitor cell population that has strong potential to contribute to valve calcification. PMID- 19218346 TI - Semicarbazide-sensitive amine oxidase/vascular adhesion protein-1 deficiency reduces leukocyte infiltration into adipose tissue and favors fat deposition. AB - Obesity is associated with low-grade inflammation and leukocyte infiltration in white adipose tissue (WAT) and is linked to diabetic complications. Semicarbazide sensitive amine oxidase, also known as vascular adhesion protein-1 (SSAO/VAP-1), is a membrane protein that is highly expressed in adipocytes and is also present on the endothelial cell surface where it is involved in leukocyte extravasation. We studied fat deposition and leukocyte infiltration in WAT of mice with a null mutation in the amine oxidase copper-containing-3 (AOC3) gene encoding SSAO/VAP 1. Both epididymal and inguinal WATs were larger in 6-month-old AOC3-KO males than in age-matched wild-type controls. However, WAT from AOC3-KO mice contained lower CD45 mRNA levels and fewer CD45(+) leukocytes. Subpopulation analyses revealed a diminished infiltration of WAT by T cells, macrophages, natural killer, and natural killer T cells. A decrease in leukocyte content in WAT was also detected in female AOC3-KO mice as early as 2 months of age, whereas increased fat mass was evident by 6 months of age. Reduced CD45(+) populations in WAT of AOC3-KO mice was not rescued by human SSAO/VAP-1 expression on adipocytes under the control of aP2, suggesting the importance of vascular AOC3 in leukocyte entrance into fat. Our results indicate that SSAO/VAP-1 is instrumental for the presence of leukocytes in WAT. Therefore, AOC3-KO mice present a unique model of mild obesity, characterized by increased WAT devoid of low-grade inflammation. PMID- 19218345 TI - Cryptococcal urease promotes the accumulation of immature dendritic cells and a non-protective T2 immune response within the lung. AB - Urease, a major virulence factor for Cryptococcus neoformans, promotes lethal meningitis/encephalitis in mice. The effect of urease within the lung, the primary site of most invasive fungal infections, is unknown. An established model of murine infection that utilizes either urease-producing (wt and ure1::URE1) or urease-deficient (ure1) strains (H99) of C. neoformans was used to characterize fungal clearance and the resultant immune response evoked by these strains within the lung. Results indicate that mice infected with urease-producing strains of C. neoformans demonstrate a 100-fold increase in fungal burden beginning 2 weeks post-infection (as compared with mice infected with urease-deficient organisms). Infection with urease-producing C. neoformans was associated with a highly polarized T2 immune response as evidenced by increases in the following: 1) pulmonary eosinophils, 2) serum IgE levels, 3) T2 cytokines (interleukin-4, -13, and -4 to interferon-gamma ratio), and 4) alternatively activated macrophages. Furthermore, the percentage and total numbers of immature dendritic cells within the lung-associated lymph nodes was markedly increased in mice infected with urease-producing C. neoformans. Collectively, these data define cryptococcal urease as a pulmonary virulence factor that promotes immature dendritic cell accumulation and a potent, yet non-protective, T2 immune response. These findings provide new insights into mechanisms by which microbial factors contribute to the immunopathology associated with invasive fungal disease. PMID- 19218347 TI - Gallbladder epithelial cells that engraft in mouse liver can differentiate into hepatocyte-like cells. AB - We tested the hypothesis that well-differentiated gallbladder epithelial cells (GBECs) are capable of engrafting and surviving in murine liver and acquire phenotypic characteristics of hepatocytes. GBECs isolated from transgenic mice that constitutively express green fluorescent protein (GFP) were either cultured before transplantation or transplanted immediately following isolation. Recipient mice with severe-combined immunodeficiency underwent retrorsine treatment and either partial hepatectomy before transplantation or carbon tetrachloride treatment following transplantation. From 1 to 4 months following transplantation, the livers of recipient mice contained discrete colonies of GFP(+) cells. Most GFP(+) cells surrounded vesicles, were epithelial cell-like in morphology, and expressed the biliary epithelial markers cytokeratin 19 and carbonic anhydrase IV. Subpopulations of GFP(+) cells resembled hepatocytes morphologically and expressed the hepatocyte-specific markers connexin-32 and hepatic nuclear factor-4alpha, but not cytokeratin 19 or carbonic anhydrase IV. At 4 months, cells in GFP(+) colonies were not actively proliferating as determined by proliferating cell nuclear antigen expression. Thus, GBECs are capable of engrafting and surviving in damaged mouse livers, and some can differentiate into cells with hepatocyte-like features. These findings suggest that environmental cues in the recipient liver are sufficient to allow a subpopulation of donor GBECs to differentiate into hepatocyte-like cells in the absence of exogenous transcriptional reprogramming. GBECs might be used as donor cells in a cell transplantation approach for the treatment of liver disease. PMID- 19218348 TI - Role of regulatory T cells in a new mouse model of experimental autoimmune myositis. AB - Polymyositis is a rare and severe inflammatory muscle disorder. Treatments are partially efficacious but have many side effects. New therapeutic approaches must be first tested in a relevant animal model. Regulatory CD4+CD25+ T cells (Tregs) have been rediscovered as a pivotal cell population in the control of autoimmunity, but the connection between polymyositis and Tregs is currently unknown. To develop a reproducible experimental autoimmune myositis model of polymyositis, mice were immunized once a week for 3 weeks with 1 mg of partially purified myosin emulsified in complete Freund's adjuvant. All mice injected with myosin and complete Freund's adjuvant developed myositis. The infiltrates were composed of CD4(+) and CD8(+) cells, as well as macrophages, but did not contain B lymphocytes. In mice that were depleted of Tregs, the myositis was more severe, as determined by quantitative scoring of muscle inflammation (2.36 +/- 0.9 vs. 1.64 +/- 0.8, P = 0.019). In contrast, injection of in vitro expanded polyclonal Tregs at the time of immunization significantly improved the disease (quantitative score of inflammation 0.87 +/- 1.06 vs. 2.4 +/- 0.67, P = 0.047). Transfer of sensitized or CD4(+)-sorted cells from the lymph nodes of experimental autoimmune myositis mice induced myositis in naive, irradiated, recipient mice. Thus, experimental autoimmune myositis is a reproducible, transferable disease in mice, both aggravated by Treg depletion and improved by polyclonal Treg injection. PMID- 19218349 TI - Fetal liver cells transplanted in utero rescue the osteopetrotic phenotype in the oc/oc mouse. AB - Autosomal recessive osteopetrosis (ARO) is a group of genetic disorders that involve defects that preclude the normal function of osteoclasts, which differentiate from hematopoietic precursors. In half of human cases, ARO is the result of mutations in the TCIRG1 gene, which codes for a subunit of the vacuolar proton pump that plays a fundamental role in the acidification of the cell-bone interface. Functional mutations of this pump severely impair the resorption of bone mineral. Although postnatal hematopoietic stem cell transplantation can partially rescue the hematological phenotype of ARO, other stigmata of the disease, such as secondary neurological and growth defects, are not reversed. For this reason, ARO is a paradigm for genetic diseases that would benefit from effective prenatal treatment. Using the oc/oc mutant mouse, a murine model whose osteopetrotic phenotype closely recapitulates human TCIRG1-dependent ARO, we report that in utero transplantation of adult bone marrow hematopoietic stem cells can correct the ARO phenotype in a limited number of mice. Here we report that in utero injection of allogeneic fetal liver cells, which include hematopoietic stem cells, into oc/oc mouse fetuses at 13.5 days post coitum produces a high level of engraftment, and the oc/oc phenotype is completely rescued in a high percentage of these mice. Therefore, oc/oc pathology appears to be particularly sensitive to this form of early treatment of the ARO genetic disorder. PMID- 19218350 TI - PASS: a program to align short sequences. AB - SUMMARY: Standard DNA alignment programs are inadequate to manage the data produced by new generation DNA sequencers. To answer this problem, we developed PASS with the objective of improving execution time and sensitivity when compared with other available programs. PASS performs fast gapped and ungapped alignments of short DNA sequences onto a reference DNA, typically a genomic sequence. It is designed to handle a huge amount of reads such as those generated by Solexa, SOLiD or 454 technologies. The algorithm is based on a data structure that holds in RAM the index of the genomic positions of 'seed' words (typically 11 and 12 bases) as well as an index of the precomputed scores of short words (typically seven and eight bases) aligned against each other. After building the genomic index, the program scans every query sequence performing three steps: (1) it finds matching seed words in the genome; (2) for every match checks the precomputed alignment of the short flanking regions; (3) if passes step 2, then it performs an exact dynamic alignment of a narrow region around the match. The performance of the program is very striking both for sensitivity and speed. For instance, gap alignment is achieved hundreds of times faster than BLAST and several times faster than SOAP, especially when gaps are allowed. Furthermore, PASS has a higher sensitivity when compared with the other available programs. AVAILABILITY AND IMPLEMENTATION: Source code and binaries are freely available for download at http://pass.cribi.unipd.it, implemented in C++and supported on Linux and Windows. PMID- 19218351 TI - Gene network reconstruction from transcriptional dynamics under kinetic model uncertainty: a case for the second derivative. AB - MOTIVATION: Measurements of gene expression over time enable the reconstruction of transcriptional networks. However, Bayesian networks and many other current reconstruction methods rely on assumptions that conflict with the differential equations that describe transcriptional kinetics. Practical approximations of kinetic models would enable inferring causal relationships between genes from expression data of microarray, tag-based and conventional platforms, but conclusions are sensitive to the assumptions made. RESULTS: The representation of a sufficiently large portion of genome enables computation of an upper bound on how much confidence one may place in influences between genes on the basis of expression data. Information about which genes encode transcription factors is not necessary but may be incorporated if available. The methodology is generalized to cover cases in which expression measurements are missing for many of the genes that might control the transcription of the genes of interest. The assumption that the gene expression level is roughly proportional to the rate of translation led to better empirical performance than did either the assumption that the gene expression level is roughly proportional to the protein level or the Bayesian model average of both assumptions. AVAILABILITY: http://www.oisb.ca points to R code implementing the methods (R Development Core Team 2004). SUPPLEMENTARY INFORMATION: http://www.davidbickel.com. PMID- 19218352 TI - CGAS: comparative genomic analysis server. AB - SUMMARY: Comparative approach is one of the most essential methods for extracting functional and evolutionary information from genomic sequences. So far, a number of sequence comparison tools have been developed, and most are either for on-site use, requiring program installation but providing a wide variety of analyses, or for the online search of user's sequences against given databases on a server. We newly devised an Asynchronous JavaScript and XML (Ajax)-based system for comparative genomic analyses, CGAS, with highly interactive interface within a browser, requiring no software installation. The current version, CGAS version 1, provides functionality for viewing similarity relationships between user's sequences, including a multiple dot plot between sequences with their annotation information. The scrollbar-less 'draggable' interface of CGAS is implemented with Google Maps API version 2. The annotation information associated with the genomic sequences compared is synchronously displayed with the comparison view. The multiple-comparison viewer is one of the unique functionalities of this system to allow the users to compare the differences between different pairs of sequences. In this viewer, the system tells orthologous correspondences between the sequences compared interactively. This web-based tool is platform-independent and will provide biologists having no computational skills with opportunities to analyze their own data without software installation and customization of the computer system. AVAILABILITY AND IMPLEMENTATION: CGAS is available at http://cgas.ist.hokudai.ac.jp/. PMID- 19218353 TI - TNF-induced activation of pulmonary microvessel endothelial cells: a role for GSK3beta. AB - The hypothesis tested was PKCalpha mediates the phosphorylation of glycogen synthetase kinase 3beta (GSK3beta) and that the GSK3beta inhibition modulates the response to tumor necrosis factor-alpha (TNF) in rat pulmonary microvessel endothelial cells (PMEC). PMEC were treated with TNF for 4.0 h (100 ng/ml) or vehicle. First, to assess the role of PKCalpha in the phosphorylation of GSK3beta (i.e., an indicator of GSK3beta inhibition), PMEC were pretreated with 1) nonsense-RNA-PKCalpha, 2) siRNA-PKCalpha, and 3) the PKC inhibitor Go6983. In the nonsense RNA-PKCalpha+TNF and TNF groups, there was increased phosphorylated GSK3beta-Ser9 that did not occur in the Go6983+TNF group. In the TNF groups, there was a significant correlation between PKCalpha protein and phosphorylated GSK3beta-Ser9 that did not occur in the groups without TNF. Second, to assess the role of GSK3beta in beta-catenin activity, PMEC were pretreated with 1) wild-type (w) GSK3beta plasmid to enhance GSK3beta activity, 2) kinase dead (kd)-GSK3beta plasmid, and 3) the GSK3beta inhibitor SB-216763. In the TNF group, there was increased unphosphorylated beta-catenin-Ser37/33 compared with the control group. In the GSK3beta-inhibited groups (i.e., SB-216763 and kdGSK3beta) +/- TNF, the unphosphorylated beta-catenin-Ser37/33 was similar to the TNF group. In the GSK3beta-enhanced group +/- TNF, the unphosphorylated beta-catenin-Ser37/33 was similar to the control. Finally, PMEC were also treated with TOPflash, a beta catenin-dependent promoter luciferase reporter, or the mutant construct FOPflash, 2 days before treatment with TNF. In the TNF group, there was an increased TOPflash/FOPflash activity ratio compared with the control group. In the GSK3beta inhibited groups (i.e., SB-216763 and kdGSK3beta) +/- TNF, the TOPflash/FOPflash activity ratio was similar to the TNF group. In the GSK3beta-enhanced group +/- TNF, the TOPflash/FOPflash activity ratio was similar to the control. The data indicate that TNF induces endothelial activation that is modulated by a PKCalpha dependent inhibition of GSK3beta. PMID- 19218354 TI - Maternal exposure to endotoxin delays alveolarization during postnatal rat lung development. AB - Maternal bacterial infections adversely affect lung development by crossing the placental barrier and infecting the developing fetus. The underlying mechanism negatively affecting pulmonary development remains unknown. Herein, we investigated whether a systemic maternal infection affects postnatal inflammation and alveolar development. Pregnant rats were injected with 2.5 mg/kg LPS on day 20 and 21 (term = 22 days). Postnatal (PN0-21) mRNA and protein expression of cytokines (IL-1beta, IL-6, IL-10, CXCL1/2, TNFalpha) and genes implicated in alveologenesis [tropoelastin, lysyl oxidase (LOX), lysyl oxidase-like (LOXL)1, tenascin-C (TNC), fibulin 5, vascular endothelial growth factor (VEGF-A), VEGF receptor (VEGFR)2, VEGFR1, platelet-derived growth factor (PDGF)A, PDGFB, and PDGFRalpha] were quantified by real-time PCR and beadlyte technology. Lung transcript and protein levels of IL-1beta, IL-6, and CXCL1/2 were significantly greater in LPS-exposed pups than those of control pups at PN0, 2, 6, 10, and 14. Bronchoalveolar lavage fluid (BALF) of LPS-exposed animals contained significantly more macrophages at PN2 and 14 than BALF of control pups. Morphometric analysis revealed that LPS-exposed animals had fewer and larger alveoli, fewer secondary septa, and decreased peripheral vessel density when compared with control pups. This morphological delay in alveolar development disappeared after PN14. Tropoelastin, LOXL1, VEGF, VEGFR2, and PDGFRalpha mRNA expression of LPS-exposed animals was significantly greater than those of control animals in PN2-14 lungs. TNC, LOX, fibulin 5, VEGFR1, PDGFA, and PDGFB expression was not affected by maternal LPS exposure. Together, the data demonstrate that maternal exposure to endotoxin results in a prolonged pulmonary inflammation postnatally, altered gene expression of molecules implicated in alveologenesis, and delayed morphological maturation of the lung. PMID- 19218357 TI - Modelling and measuring electromechanical coupling in the rat heart. AB - Tension-dependent binding of Ca(2+) to troponin C in the cardiac myocyte has been shown to play an important role in the regulation of Ca(2+) and the activation of tension development. The significance of this regulatory mechanism is quantified experimentally by the quantity of Ca(2+) released following a rapid change in the muscle length. Using a computational, coupled, electromechanics cell model, we have confirmed that the tension dependence of Ca(2+) binding to troponin C, rather than cross-bridge kinetics or the rate of Ca(2+) uptake by the sarcoplasmic reticulum, determines the quantity of Ca(2+) released following a length step. This cell model has been successfully applied in a continuum model of the papillary muscle to analyse experimental data, suggesting the tension dependent binding of Ca(2+) to troponin C as the likely pathway through which the effects of localized impaired tension generation alter the Ca(2+) transient. These experimental results are qualitatively reproduced using a three-dimensional coupled electromechanics model. Furthermore, the model predicts that changes in the Ca(2+) transient in the viable myocardium surrounding the impaired region are amplified in the absence of tension-dependent binding of Ca(2+) to troponin C. PMID- 19218355 TI - Recent advances in genetic predisposition to clinical acute lung injury. AB - It has been well established that acute lung injury (ALI), and the more severe presentation of acute respiratory distress syndrome (ARDS), constitute complex traits characterized by a multigenic and multifactorial etiology. Identification and validation of genetic variants contributing to disease susceptibility and severity has been hampered by the profound heterogeneity of the clinical phenotype and the role of environmental factors, which includes treatment, on outcome. The critical nature of ALI and ARDS, compounded by the impact of phenotypic heterogeneity, has rendered the amassing of sufficiently powered studies especially challenging. Nevertheless, progress has been made in the identification of genetic variants in select candidate genes, which has enhanced our understanding of the specific pathways involved in disease manifestation. Identification of novel candidate genes for which genetic association studies have confirmed a role in disease has been greatly aided by the powerful tool of high-throughput expression profiling. This article will review these studies to date, summarizing candidate genes associated with ALI and ARDS, acknowledging those that have been replicated in independent populations, with a special focus on the specific pathways for which candidate genes identified so far can be clustered. PMID- 19218358 TI - The training stimulus experienced by the leg muscles during cycling in humans. AB - Considerable variability exists between people in their health- and performance related adaptations to conventional endurance training. We hypothesized that some of this variability might be due to differences in the training stimulus received by the working muscles. In 71 young sedentary women we observed large variations in the ratio of one-leg cycling muscle aerobic capacity (V(O2peak)) to two-leg cycling whole-body maximal oxygen uptake (V(O2max); Ratio(1:2); range 0.58-0.96). The variability in Ratio(1:2) was primarily due to differences between people in one-leg V(O2peak) (r = 0.71, P < 0.0005) and was not related to two-leg V(O2max) (r = 0.15, P = 0.209). Magnetic resonance imaging (n = 30) and muscle biopsy sampling (n = 20) revealed that one-leg V(O2peak) was mainly determined by muscle volume (r = 0.73, P < 0.0005) rather than muscle fibre type or oxidative capacity. A high one-leg V(O2peak) was associated with favourable lipoprotein profiles (P = 0.033, n = 24) but this was not the case for two-leg V(O2max). Calculations based on these data suggest that conventional two-leg exercise at 70% V(O2max) requires subjects with the lowest Ratio(1:2) to work their legs at 60% of single-leg V(O2peak), whilst those with the highest Ratio(1:2) work their legs at only 36% of maximum. It was concluded that endurance training carried out according to current guidelines will result in highly variable training stimuli for the leg muscles and variable magnitudes of adaptation. These conclusions have implications for the prescription of exercise to improve health and for investigations into the genetic basis of muscle adaptations. PMID- 19218356 TI - Cardiovascular and autonomic phenotype of db/db diabetic mice. AB - The db/db mice serve as a good model for type 2 diabetes characterized by hyperinsulinaemia and progressive hyperglycaemia. There are limited and conflicting data on the cardiovascular changes in this model. The aim of the present study was to characterize the cardiovascular and autonomic phenotype of male db/db mice and evaluate the role of angiotensin II AT(1) receptors. Radiotelemetry was used to monitor 24 h blood pressure (BP) in mice for 8 weeks. Parameters measured were mean arterial pressure (MAP), heart rate (HR) and their variabilities. In 8-week-old db/db mice, the MAP and BP circadian rhythms were not different from age-matched control mice, while HR and locomotor activity were decreased. With ageing, MAP gradually increased in db/db mice, and the 12 h light values did not dip significantly from the 12 h dark periods. In 14-week-old mice, MAP was increased during light (101 +/- 1 versus 117 +/- 2 mmHg, P < 0.01; control versus db/db mice) and dark phases (110 +/- 1.7 versus 121 +/- 3.1 mmHg, P < 0.01; control versus db/db mice). This increase in MAP was associated with a significant increase in plasma angiotensin-converting enzyme activity and angiotensin II levels. Chronic treatment with losartan (10 mg kg(-1) day(-1)) blocked the increase in MAP in db/db mice, with no effect in control animals. Spectral analysis was used to monitor autonomic cardiovascular function. The circadian rhythm observed in systolic arterial pressure variance and its low frequency component in control mice was absent in db/db mice. There were no changes in HR variability and spontaneous baroreflex sensitivity between control and db/db mice. The results document an age-related increase in MAP in db/db mice, which can be reduced by antagonism of angiotensin II AT(1) receptors, and alterations in autonomic balance and components of the renin-angiotensin system. PMID- 19218359 TI - Differing roles for members of the phospholipase A2 superfamily in experimental autoimmune encephalomyelitis. AB - The phospholipase A(2) (PLA(2)) superfamily hydrolyzes phospholipids to release free fatty acids and lysophospholipids, some of which can mediate inflammation and demyelination, hallmarks of the CNS autoimmune disease multiple sclerosis. The expression of two of the intracellular PLA(2)s (cPLA(2) GIVA and iPLA(2) GVIA) and two of the secreted PLA(2)s (sPLA(2) GIIA and sPLA(2) GV) are increased in different stages of experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. We show using small molecule inhibitors, that cPLA(2) GIVA plays a role in the onset, and iPLA(2) GVIA in the onset and progression of EAE. We also show a potential role for sPLA(2) in the later remission phase. These studies demonstrate that selective inhibition of iPLA(2) can ameliorate disease progression when treatment is started before or after the onset of symptoms. The effects of these inhibitors on lesion burden, chemokine and cytokine expression as well as on the lipid profile provide insights into their potential modes of action. iPLA(2) is also expressed by macrophages and other immune cells in multiple sclerosis lesions. Our results therefore suggest that iPLA(2) might be an excellent target to block for the treatment of CNS autoimmune diseases, such as multiple sclerosis. PMID- 19218360 TI - Intramolecular electron transfer processes in Cu(B)-deficient cytochrome bo studied by pulse radiolysis. AB - The Escherichia coli cytochrome bo is a heme-copper terminal ubiquinol oxidase, and functions as a redox-driven proton pump. We applied pulse radiolysis technique for studying the one-electron reduction processes in the Cu(B) deficient mutant, His333Ala. We found that the Cu(B) deficiency suppressed the heme b-to-heme o electron transfer two order of the magnitude (4.0 x 10(2) s( 1)), as found for the wild-type enzyme in the presence of 1 mM KCN (3.0 x 10(2) s(-1)). Potentiometric analysis of the His333Ala mutant revealed the 40 mV decrease in the E(m) value for low-spin heme b and the 160 mV increase in the E(m) value of high-spin heme o. Our results indicate that Cu(B) not only serves as one-electron donor to the bound dioxygen upon the O-O bond cleavage, but also facilitates dioxygen reduction at the heme-copper binuclear centre by modulating the E(m) value of heme o through magnetic interactions. And the absence of a putative OH(-) bound to Cu(B) seems not to affect the uptake of the first chemical proton via K-channel in the His333Ala mutant. PMID- 19218361 TI - Roles for auxin, cytokinin, and strigolactone in regulating shoot branching. AB - Many processes have been described in the control of shoot branching. Apical dominance is defined as the control exerted by the shoot tip on the outgrowth of axillary buds, whereas correlative inhibition includes the suppression of growth by other growing buds or shoots. The level, signaling, and/or flow of the plant hormone auxin in stems and buds is thought to be involved in these processes. In addition, RAMOSUS (RMS) branching genes in pea (Pisum sativum) control the synthesis and perception of a long-distance inhibitory branching signal produced in the stem and roots, a strigolactone or product. Auxin treatment affects the expression of RMS genes, but it is unclear whether the RMS network can regulate branching independently of auxin. Here, we explore whether apical dominance and correlative inhibition show independent or additive effects in rms mutant plants. Bud outgrowth and branch lengths are enhanced in decapitated and stem-girdled rms mutants compared with intact control plants. This may relate to an RMS independent induction of axillary bud outgrowth by these treatments. Correlative inhibition was also apparent in rms mutant plants, again indicating an RMS independent component. Treatments giving reductions in RMS1 and RMS5 gene expression, auxin transport, and auxin level in the main stem were not always sufficient to promote bud outgrowth. We suggest that this may relate to a failure to induce the expression of cytokinin biosynthesis genes, which always correlated with bud outgrowth in our treatments. We present a new model that accounts for apical dominance, correlative inhibition, RMS gene action, and auxin and cytokinin and their interactions in controlling the progression of buds through different control points from dormancy to sustained growth. PMID- 19218362 TI - Engineered polyamine catabolism preinduces tolerance of tobacco to bacteria and oomycetes. AB - Polyamine oxidase (PAO) catalyzes the oxidative catabolism of spermidine and spermine, generating hydrogen peroxide. In wild-type tobacco (Nicotiana tabacum 'Xanthi') plants, infection by the compatible pathogen Pseudomonas syringae pv tabaci resulted in increased PAO gene and corresponding PAO enzyme activities; polyamine homeostasis was maintained by induction of the arginine decarboxylase pathway and spermine was excreted into the apoplast, where it was oxidized by the enhanced apoplastic PAO, resulting in higher hydrogen peroxide accumulation. Moreover, plants overexpressing PAO showed preinduced disease tolerance against the biotrophic bacterium P. syringae pv tabaci and the hemibiotrophic oomycete Phytophthora parasitica var nicotianae but not against the Cucumber mosaic virus. Furthermore, in transgenic PAO-overexpressing plants, systemic acquired resistance marker genes as well as a pronounced increase in the cell wall-based defense were found before inoculation. These results reveal that PAO is a nodal point in a specific apoplast-localized plant-pathogen interaction, which also signals parallel defense responses, thus preventing pathogen colonization. This strategy presents a novel approach for producing transgenic plants resistant to a broad spectrum of plant pathogens. PMID- 19218363 TI - Inhibition of tobacco mosaic virus movement by expression of an actin-binding protein. AB - The tobacco mosaic virus (TMV) movement protein (MP) required for the cell-to cell spread of viral RNA interacts with the endoplasmic reticulum (ER) as well as with the cytoskeleton during infection. Whereas associations of MP with ER and microtubules have been intensely investigated, research on the role of actin has been rather scarce. We demonstrate that Nicotiana benthamiana plants transgenic for the actin-binding domain 2 of Arabidopsis (Arabidopsis thaliana) fimbrin (AtFIM1) fused to green fluorescent protein (ABD2:GFP) exhibit a dynamic ABD2:GFP labeled actin cytoskeleton and myosin-dependent Golgi trafficking. These plants also support the movement of TMV. In contrast, both myosin-dependent Golgi trafficking and TMV movement are dominantly inhibited when ABD2:GFP is expressed transiently. Inhibition is mediated through binding of ABD2:GFP to actin filaments, since TMV movement is restored upon disruption of the ABD2:GFP-labeled actin network with latrunculin B. Latrunculin B shows no significant effect on the spread of TMV infection in either wild-type plants or ABD2:GFP transgenic plants under our treatment conditions. We did not observe any binding of MP along the length of actin filaments. Collectively, these observations demonstrate that TMV movement does not require an intact actomyosin system. Nevertheless, actin binding proteins appear to have the potential to exert control over TMV movement through the inhibition of myosin-associated protein trafficking along the ER membrane. PMID- 19218364 TI - CIRCADIAN CLOCK ASSOCIATED1 and LATE ELONGATED HYPOCOTYL function synergistically in the circadian clock of Arabidopsis. AB - The circadian clock is an endogenous mechanism that coordinates biological processes with daily and seasonal changes in the environment. Heterodimerization of central clock components is an important way of controlling clock function in several different circadian systems. CIRCADIAN CLOCK ASSOCIATED1 (CCA1) and LATE ELONGATED HYPOCOTYL (LHY) are Myb-related proteins that function in or close to the central oscillator in Arabidopsis (Arabidopsis thaliana). Single mutants of cca1 and lhy have a phenotype of short-period rhythms. cca1 lhy double mutants show an even shorter period phenotype than the cca1 single mutant, suggesting that CCA1 and LHY are only partially functionally redundant. To determine whether CCA1 and LHY act in parallel or synergistically in the circadian clock, we examined their expression in both light-grown and etiolated seedlings. We have shown that LHY and CCA1 bind to the same region of the promoter of a Light harvesting chlorophyll a/b protein (Lhcb, also known as CAB). CCA1 and LHY can form homodimers, and they also colocalize in the nucleus and heterodimerize in vitro and in vivo. In Arabidopsis, CCA1 and LHY physically interact in a manner independent of photoperiod. Moreover, results from gel filtration chromatography indicate that CCA1 and LHY are present in the same large complex in plants. Taken together, these results imply that CCA1 and LHY function synergistically in regulating circadian rhythms of Arabidopsis. PMID- 19218365 TI - Homology modeling and site-directed mutagenesis reveal catalytic key amino acids of 3beta-hydroxysteroid-dehydrogenase/C4-decarboxylase from Arabidopsis. AB - Sterols become functional only after removal of the two methyl groups at C4 by a membrane-bound multienzyme complex including a 3beta-hydroxysteroid dehydrogenase/C4-decarboxylase (3betaHSD/D). We recently identified Arabidopsis (Arabidopsis thaliana) 3betaHSD/D as a bifunctional short-chain dehydrogenase/reductase protein. We made use of three-dimensional homology modeling to identify key amino acids involved in 4alpha-carboxy-sterol and NAD binding and catalysis. Key amino acids were subjected to site-directed mutagenesis, and the mutated enzymes were expressed and assayed both in vivo and in vitro in an erg26 yeast strain defective in 3betaHSD/D. We show that tyrosine 159 and lysine-163, which are oriented near the 3beta-hydroxyl group of the substrate in the model, are essential for the 3betaHSD/D activity, consistent with their involvement in the initial dehydrogenation step of the reaction. The essential arginine-326 residue is predicted to form a salt bridge with the 4alpha carboxyl group of the substrate, suggesting its involvement both in substrate binding and in the decarboxylation step. The essential aspartic acid-39 residue is in close contact with the hydroxyl groups of the adenosine-ribose ring of NAD+, in good agreement with the strong preference of 3betaHSD/D for NAD+. Data obtained with serine-133 mutants suggest close proximity between the serine-133 residue and the C4beta domain of the bound sterol. Based on these data, we propose a tentative mechanism for 3betaHSD/D activity. This study provides, to our knowledge, the first data on the three-dimensional molecular interactions of an enzyme of the postoxidosqualene cyclase sterol biosynthesis pathway with its substrate. The implications of our findings for studying the roles of C4 alkylated sterol precursors in plant development are discussed. PMID- 19218366 TI - Niflumic acid alters gating of HCN2 pacemaker channels by interaction with the outer region of S4 voltage sensing domains. AB - Niflumic acid, 2-[[3-(trifluoromethyl)phenyl]amino]pyridine-3-carboxylic acid (NFA), is a nonsteroidal anti-inflammatory drug that also blocks or modifies the gating of many ion channels. Here, we investigated the effects of NFA on hyperpolarization-activated cyclic nucleotide-gated cation (HCN) pacemaker channels expressed in X. laevis oocytes using site-directed mutagenesis and the two-electrode voltage-clamp technique. Extracellular NFA acted rapidly and caused a slowing of activation and deactivation and a hyperpolarizing shift in the voltage dependence of HCN2 channel activation (-24.5 +/- 1.2 mV at 1 mM). Slowed channel gating and reduction of current magnitude was marked in oocytes treated with NFA, while clamped at 0 mV but minimal in oocytes clamped at -100 mV, indicating the drug preferentially interacts with channels in the closed state. NFA at 0.1 to 3 mM shifted the half-point for channel activation in a concentration-dependent manner, with an EC(50) of 0.54 +/- 0.068 mM and a predicted maximum shift of -38 mV. NFA at 1 mM also reduced maximum HCN2 conductance by approximately 20%, presumably by direct block of the pore. The rapid onset and state-dependence of NFA-induced changes in channel gating suggests an interaction with the extracellular region of the S4 transmembrane helix, the primary voltage-sensing domain of HCN2. Neutralization (by mutation to Gln) of any three of the outer four basic charged residues in S4, but not single mutations, abrogated the NFA-induced shift in channel activation. We conclude that NFA alters HCN2 gating by interacting with the extracellular end of the S4 voltage sensor domains. PMID- 19218367 TI - Time-resolved photolabeling of the nicotinic acetylcholine receptor by [3H]azietomidate, an open-state inhibitor. AB - Azietomidate is a photoreactive analog of the general anesthetic etomidate that acts as a nicotinic acetylcholine receptor (nAChR) noncompetitive antagonist. We used rapid perfusion electrophysiological techniques to characterize the state dependence and kinetics of azietomidate inhibition of Torpedo californica nAChRs and time-resolved photolabeling to identify the nAChR binding sites occupied after exposure to [(3)H]azietomidate and agonist for 50 ms (open state) or at equilibrium (desensitized state). Azietomidate acted primarily as an open channel inhibitor characterized by a bimolecular association rate constant of k(+) = 5 x 10(5) M(-1) s(-1) and a dissociation rate constant of <3s(-1). Azietomidate at 10 microM, when perfused with acetylcholine (ACh), inhibited the ACh response by approximately 50% after 50 ms; when preincubated for 10 s, it decreased the peak initial response by approximately 15%. Comparison of the kinetics of recovery of ACh responses after exposure to ACh and azietomidate or to ACh alone indicated that at subsecond times, azietomidate inhibited nAChRs without enhancing the kinetics of agonist-induced desensitization. In nAChRs frozen after 50-ms exposure to agonist and [(3)H]azietomidate, amino acids were photolabeled in the ion channel [position M2-20 (alphaGlu-262, betaAsp-268, deltaGln-276)], in deltaM1 (deltaCys-236), and in alphaMA/alphaM4 (alphaGlu-390, alphaCys-412) that were also photolabeled in nAChRs in the equilibrium desensitized state at approximately half the efficiency. These results identify azietomidate binding sites at the extracellular end of the ion channel, in the delta subunit helix bundle, and in the nAChR cytoplasmic domain that seem similar in structure and accessibility in the open and desensitized states of the nAChR. PMID- 19218369 TI - Hypotension in obstetric spinal anaesthesia: a lesson from pre-eclampsia. PMID- 19218370 TI - Red cell transfusion in elective cardiac surgery patients: where do we go from here? PMID- 19218371 TI - Smoking and alcohol intervention before surgery: evidence for best practice. AB - Smoking and hazardous drinking are common and important risk factors for an increased rate of complications after surgery. The underlying pathophysiological mechanisms include organic dysfunctions that can recover with abstinence. Abstinence starting 3-8 weeks before surgery will significantly reduce the incidence of several serious postoperative complications, such as wound and cardiopulmonary complications and infections. However, this intervention must be intensive to obtain sufficient effect on surgical complications. All patients presenting for surgery should be questioned regarding smoking and hazardous drinking, and interventions appropriate for the surgical setting applied. PMID- 19218372 TI - General anaesthesia for intralesional bleomycin therapy of vascular malformations: initial 3 yr experience. PMID- 19218373 TI - Spinal anaesthesia for elective surgery. PMID- 19218374 TI - Target controlled infusion of opioids for bariatric surgery and morphine loading dose. PMID- 19218375 TI - Sudden hypoxaemia in a critically ill adult patient: anomalous drainage of the upper left pulmonary vein. PMID- 19218376 TI - A new type of coronary artery stent. PMID- 19218377 TI - Short message service for outpatient data collection. PMID- 19218378 TI - Subspecies IIIa and IIIb Salmonellae are defective for colonization of murine models of salmonellosis compared to Salmonella enterica subsp. I serovar typhimurium. AB - Non-subspecies I salmonellae are commensals of cold-blooded vertebrates and cause sporadic disease in mammals. The reasons why non-subspecies I salmonellae do not circulate in populations of warm-blooded vertebrates, but instead only cause occasional disease in this niche, are unknown. We examined the ability of Salmonella enterica subsp. IIIa (subsp. arizonae) and subsp. IIIb (subsp. diarizonae) isolates to grow competitively with subspecies I (serovar Typhimurium) ATCC 14028 in vitro, to colonize Salmonella-sensitive BALB/c mice, and to persist in the intestine of Salmonella-resistant CBA/J mice in competitive infections. Subspecies IIIa had severely reduced intestinal colonization, intestinal persistence, and systemic spread in mice. Subspecies IIIa is nonmotile on swarming agar and thus may also have reduced motility under viscous conditions in vivo. Surprisingly, subspecies IIIb colonizes the intestinal tract of BALB/c mice normally yet does not spread systemically. Subspecies IIIb colonization of the intestine of CBA/J mice is reduced late in infection. In order to understand why these isolates do not colonize systemic sites, we determined that subspecies IIIa and IIIb are not internalized well and do not replicate in J774-A.1 murine macrophages, despite normal adherence to these cells. We further show that selected effectors of both type III secretion systems 1 and 2 are secreted by subspecies IIIa and IIIb in vitro but that each of these isolates secretes a different combination of effectors. We outline the phenotypic differences between these subspecies and subspecies I and provide a possible explanation for the inability of these strains to spread systemically in murine models. PMID- 19218379 TI - Functional genomics of Enterococcus faecalis: multiple novel genetic determinants for biofilm formation in the core genome. AB - The ability of Enterococcus faecalis to form robust biofilms on host tissues and on abiotic surfaces such as catheters likely plays a major role in the pathogenesis of opportunistic antibiotic-resistant E. faecalis infections and in the transfer of antibiotic resistance genes. We have carried out a comprehensive analysis of genetic determinants of biofilm formation in the core genome of E. faecalis. Here we describe 68 genetic loci predicted to be involved in biofilm formation that were identified by recombinase in vivo expression technology (RIVET); most of these genes have not been studied previously. Differential expression of a number of these determinants during biofilm growth was confirmed by quantitative reverse transcription-PCR, and genetic complementation studies verified a role in biofilm formation for several candidate genes. Of particular interest was genetic locus EF1809, predicted to encode a regulatory protein of the GntR family. We isolated 14 independent nonsibling clones containing the putative promoter region for this gene in the RIVET screen; EF1809 also showed the largest increase in expression during biofilm growth of any of the genes tested. Since an in-frame deletion of EF1809 resulted in a severe biofilm defect that could be complemented by the cloned wild-type gene, we have designated EF1809 ebrA (enterococcal biofilm regulator). Most of the novel genetic loci identified in our studies are highly conserved in gram-positive bacterial pathogens and may thus constitute a pool of uncharacterized genes involved in biofilm formation that may be useful targets for drug discovery. PMID- 19218380 TI - SigmaS controls multiple pathways associated with intracellular multiplication of Legionella pneumophila. AB - Legionella pneumophila is the causative agent of the severe and potentially fatal pneumonia Legionnaires' disease. L. pneumophila is able to replicate within macrophages and protozoa by establishing a replicative compartment in a process that requires the Icm/Dot type IVB secretion system. The signals and regulatory pathways required for Legionella infection and intracellular replication are poorly understood. Mutation of the rpoS gene, which encodes sigma(S), does not affect growth in rich medium but severely decreases L. pneumophila intracellular multiplication within protozoan hosts. To gain insight into the intracellular multiplication defect of an rpoS mutant, we examined its pattern of gene expression during exponential and postexponential growth. We found that sigma(S) affects distinct groups of genes that contribute to Legionella intracellular multiplication. We demonstrate that rpoS mutants have a functional Icm/Dot system yet are defective for the expression of many genes encoding Icm/Dot-translocated substrates. We also show that sigma(S) affects the transcription of the cpxR and pmrA genes, which encode two-component response regulators that directly affect the transcription of Icm/Dot substrates. Our characterization of the L. pneumophila small RNA csrB homologs, rsmY and rsmZ, introduces a link between sigma(S) and the posttranscriptional regulator CsrA. We analyzed the network of sigma(S)-controlled genes by mutational analysis of transcriptional regulators affected by sigma(S). One of these, encoding the L. pneumophila arginine repressor homolog gene, argR, is required for maximal intracellular growth in amoebae. These data show that sigma(S) is a key regulator of multiple pathways required for L. pneumophila intracellular multiplication. PMID- 19218381 TI - Role of FimW, FimY, and FimZ in regulating the expression of type i fimbriae in Salmonella enterica serovar Typhimurium. AB - Type I fimbriae in Salmonella enterica serovar Typhimurium are surface appendages that facilitate binding to eukaryotic cells. Expression of the fim gene cluster is known to be regulated by three proteins--FimW, FimY, and FimZ--and a tRNA encoded by fimU. In this work, we investigated how these proteins and tRNA coordinately regulate fim gene expression. Our results indicate that FimY and FimZ independently activate the P(fimA) promoter which controls the expression of the fim structural genes. FimY and FimZ were also found to strongly activate each other's expression and weakly activate their own expression. FimW was found to negatively regulate fim gene expression by repressing transcription from the P(fimY) promoter, independent of FimY or FimZ. Moreover, FimW and FimY interact within a negative feedback loop, as FimY was found to activate the P(fimW) promoter. In the case of fimU, the expression of this gene was not found to be regulated by FimW, FimY, or FimZ. We also explored the effect of fim gene expression on Salmonella pathogenicity island 1 (SPI1). Our results indicate that FimZ alone is able to enhance the expression of hilE, a known repressor of SPI1 gene expression. Based on our results, we were able to propose an integrated model for the fim gene circuit. As this model involves a combination of positive and negative feedback, we hypothesized that the response of this circuit may be bistable and thus a possible mechanism for phase variation. However, we found that the response was continuous and not bistable. PMID- 19218382 TI - Nanoscale structural and mechanical properties of nontypeable Haemophilus influenzae biofilms. AB - Nontypeable Haemophilus influenzae (NTHI) bacteria are commensals in the human nasopharynx, as well as pathogens associated with a spectrum of acute and chronic infections. Two important factors that influence NTHI pathogenicity are their ability to adhere to human tissue and their ability to form biofilms. Extracellular polymeric substances (EPS) and bacterial appendages such as pili critically influence cell adhesion and intercellular cohesion during biofilm formation. Structural components in the outer cell membrane, such as lipopolysaccharides, also play a fundamental role in infection of the host organism. In spite of their importance, these pathogenic factors are not yet well characterized at the nanoscale. Here, atomic force microscopy (AFM) was used in aqueous environments to visualize structural details, including probable Hif-type pili, of live NTHI bacteria at the early stages of biofilm formation. Using single-molecule AFM-based spectroscopy, the molecular elasticities of lipooligosaccharides present on NTHI cell surfaces were analyzed and compared between two strains (PittEE and PittGG) with very different pathogenicity profiles. Furthermore, the stiffness of single cells of both strains was measured and subsequently their turgor pressure was estimated. PMID- 19218383 TI - The 3'-to-5' exoribonuclease (encoded by HP1248) of Helicobacter pylori regulates motility and apoptosis-inducing genes. AB - The human gastric pathogen Helicobacter pylori has many virulence factors involved in pathogenesis, but the mechanisms regulating these virulence factors are not yet fully understood. In this study, we cloned HP1248, which is similar in sequence to Escherichia coli vacB, which was previously shown to be associated with the expression of virulence in Shigella and enteroinvasive E. coli. E. coli vacB encodes RNase R. RNase R is involved in the posttranscriptional regulation of mRNA stability. By global transcriptional microarray profiling of an H. pylori HP1248 deletion mutant, we defined six virulence-related genes which were posttranscriptionally downregulated by HP1248, including the motility-related genes HP1192 and flaB, the chemotaxis-related gene cheY, and the apoptosis inducing genes HP0175, cagA, and gtt. In this study, recombinant HP1248 protein expressed in E. coli showed 3'-to-5' exoribonuclease activity. Motility and apoptosis induction were increased in the H. pylori HP1248 deletion mutant. We also showed that HP1192 is associated with H. pylori motility, possibly through HP1248 regulation. Further, we suggested and studied the possible mechanisms of this specific regulation of virulent genes by HP1248. In addition, the expression level of HP1248 mRNA changed dramatically in response to a variety of altered environmental conditions, including pH and temperature. Hence, HP1248 in H. pylori seems to play a role in environmental sensing and in regulation of virulent phenotypes, such as motility and host apoptosis induction. PMID- 19218384 TI - ClpXP degrades SsrA-tagged proteins in Streptococcus pneumoniae. AB - Bacterial proteins that are abnormally truncated due to incomplete mRNA or the presence of rare codons are extended by an SsrA tag during ribosome rescue in a trans-translation process important for maintaining protein quality. In Escherichia coli, the SsrA-tagged proteins become the target of the Tsp, Lon, FtsH, ClpXP, and ClpAP proteases. Here we show that degradation of model SsrA tagged proteins in Streptococcus pneumoniae depends primarily or exclusively on ClpXP in vivo. In addition, we show the E. coli SsrA tag is also a target of S. pneumoniae ClpXP in vivo, even though the N-terminal portions of the tags differ significantly between the two species, suggesting there may be no adaptor protein for SsrA in S. pneumoniae. PMID- 19218385 TI - Regulation of igaA and the Rcs system by the MviA response regulator in Salmonella enterica. AB - IgaA is a membrane protein that prevents overactivation of the Rcs regulatory system in enteric bacteria. Here we provide evidence that igaA is the first gene in a sigma(70)-dependent operon of Salmonella enterica serovar Typhimurium that also includes yrfG, yrfH, and yrfI. We also show that the Lon protease and the MviA response regulator participate in regulation of the igaA operon. Our results indicate that MviA regulates igaA transcription in an RpoS-dependent manner, but the results also suggest that MviA may regulate RcsB activation in an RpoS- and IgaA-independent manner. PMID- 19218386 TI - Differential expression of sigH paralogs during growth and under different stress conditions in Mycobacterium smegmatis. AB - SigH regulates a transcriptional network that responds to heat and oxidative stress in mycobacteria. Seven sigH paralogs are reported to exist in the Mycobacterium smegmatis genome. A comprehensive real-time reverse transcriptase PCR analysis during different stages of growth and upon exposure to various stress conditions and antimycobacterial compounds showed differential expression of sigH paralogs during stationary phase and severalfold increases in the levels of transcription of sigH1, sigH4, sigH5, sigH6, and sigH7 under specific stress conditions. PMID- 19218387 TI - An Aeromonas caviae genomic island is required for both O-antigen lipopolysaccharide biosynthesis and flagellin glycosylation. AB - Aeromonas caviae Sch3N possesses a small genomic island that is involved in both flagellin glycosylation and lipopolysaccharide (LPS) O-antigen biosynthesis. This island appears to have been laterally acquired as it is flanked by insertion element-like sequences and has a much lower G+C content than the average aeromonad G+C content. Most of the gene products encoded by the island are orthologues of proteins that have been shown to be involved in pseudaminic acid biosynthesis and flagellin glycosylation in both Campylobacter jejuni and Helicobacter pylori. Two of the genes, lst and lsg, are LPS specific as mutation of them results in the loss of only a band for the LPS O-antigen. Lsg encodes a putative Wzx flippase, and mutation of Lsg affects only LPS; this finding supports the notion that flagellin glycosylation occurs within the cell before the flagellins are exported and assembled and not at the surface once the sugar has been exported. The proteins encoded by flmA, flmB, neuA, flmD, and neuB are thought to make up a pseudaminic acid biosynthetic pathway, and mutation of any of these genes resulted in the loss of motility, flagellar expression, and a band for the LPS O-antigen. Furthermore, pseudaminic acid was shown to be present on both flagellin subunits that make up the polar flagellum filament, to be present in the LPS O-antigen of the A. caviae wild-type strain, and to be absent from the A. caviae flmD mutant strain. PMID- 19218388 TI - Constitutive mutations in the Escherichia coli AraC protein. AB - The Escherichia coli AraC protein represses and induces the araBAD operon in response to the absence or presence of l-arabinose. Constitutive mutations in the AraC gene no longer require the presence of l-arabinose to convert AraC from its repressing to its inducing state. Such mutations were isolated directly by virtue of their constitutivity or by their resistance to the nonmetabolizable arabinose analog, d-fucose. The majority of the constitutive mutations lie within the same residues of the N-terminal regulatory arm of AraC. Two, however, were found in the core of the dimerization domain. As predicted by the light switch mechanism of AraC, constitutive mutations increase the susceptibility of the N-terminal arms to digestion by trypsin or chymotrypsin, suggesting that these mutations weaken or disrupt the arm structure required for repression by AraC. Fluorescence, circular dichroism, and cysteine reactivity measurements show that the constitutive mutations in the core of the dimerization domain lead to a weakening of the support for the arms and reduce the stability of the minus arabinose arm structure. These mutations also weaken the interaction between the two-helix bundle and the beta-barrel subdomains of the dimerization domain and reduce the structural stability of the beta-barrels. PMID- 19218389 TI - SleC is essential for cortex peptidoglycan hydrolysis during germination of spores of the pathogenic bacterium Clostridium perfringens. AB - Clostridial spore germination requires degradation of the spore's peptidoglycan (PG) cortex by cortex-lytic enzymes (CLEs), and two Clostridium perfringens CLEs, SleC and SleM, degrade cortex PG in vitro. We now find that only SleC is essential for cortex hydrolysis and viability of C. perfringens spores. C. perfringens sleC spores did not germinate completely with nutrients, KCl, or a 1:1 chelate of Ca(2+) and dipicolinic acid (Ca-DPA), and the colony-forming efficiency of sleC spores was 10(3)-fold lower than that of wild-type spores. However, sleC spores incubated with various germinants released most of their DPA, although slower than wild-type or sleM spores, and DPA release from sleC sleM spores was very slow. In contrast, germination and viability of sleM spores were similar to that of wild-type spores, although sleC sleM spores had 10(5) fold-lower viability. These results allow the following conclusions about C. perfringens spore germination: (i) SleC is essential for cortex hydrolysis; (ii) although SleM can degrade cortex PG in vitro, this enzyme is not essential; (iii) action of SleC alone or with SleM can accelerate DPA release; and (iv) Ca-DPA does not trigger spore germination by activation of CLEs. PMID- 19218391 TI - Characterization of a Mesorhizobium loti alpha-type carbonic anhydrase and its role in symbiotic nitrogen fixation. AB - Carbonic anhydrase (CA) (EC 4.2.1.1) is a widespread enzyme catalyzing the reversible hydration of CO(2) to bicarbonate, a reaction that participates in many biochemical and physiological processes. Mesorhizobium loti, the microsymbiont of the model legume Lotus japonicus, possesses on the symbiosis island a gene (msi040) encoding an alpha-type CA homologue, annotated as CAA1. In the present work, the CAA1 open reading frame from M. loti strain R7A was cloned, expressed, and biochemically characterized, and it was proven to be an active alpha-CA. The biochemical and physiological roles of the CAA1 gene in free-living and symbiotic rhizobia were examined by using an M. loti R7A disruption mutant strain. Our analysis revealed that CAA1 is expressed in both nitrogen-fixing bacteroids and free-living bacteria during growth in batch cultures, where gene expression was induced by increased medium pH. L. japonicus plants inoculated with the CAA1 mutant strain showed no differences in top-plant traits and nutritional status but consistently formed a higher number of nodules exhibiting higher fresh weight, N content, nitrogenase activity, and delta(13)C abundance. Based on these results, we propose that although CAA1 is not essential for nodule development and symbiotic nitrogen fixation, it may participate in an auxiliary mechanism that buffers the bacteroid periplasm, creating an environment favorable for NH(3) protonation, thus facilitating its diffusion and transport to the plant. In addition, changes in the nodule delta(13)C abundance suggest the recycling of at least part of the HCO(3)(-) produced by CAA1. PMID- 19218390 TI - Borrelia burgdorferi bba74 is expressed exclusively during tick feeding and is regulated by both arthropod- and mammalian host-specific signals. AB - Although BBA74 initially was described as a 28-kDa virulence-associated outer membrane-spanning protein with porin-like function, subsequent studies revealed that it is periplasmic and downregulated in mammalian host-adapted spirochetes. To further elucidate the role of this protein in the Borrelia burgdorferi tick mammal cycle, we conducted a thorough examination of its expression profile in comparison with the profiles of three well-characterized, differentially expressed borrelial genes (ospA, ospC, and ospE) and their proteins. In vitro, transcripts for bba74 were expressed at 23 degrees C and further enhanced by a temperature shift (37 degrees C), whereas BBA74 protein diminished at elevated temperatures; in contrast, neither transcript nor protein was expressed by spirochetes grown in dialysis membrane chambers (DMCs). Primer extension of wild type B. burgdorferi grown in vitro, in conjunction with expression analysis of DMC-cultivated wild-type and rpoS mutant spirochetes, revealed that, like ospA, bba74 is transcribed by sigma(70) and is subject to RpoS-mediated repression within the mammalian host. A series of experiments utilizing wild-type and rpoS mutant spirochetes was conducted to determine the transcriptional and translational profiles of bba74 during the tick-mouse cycle. Results from these studies revealed (i) that bba74 is transcribed by sigma(70) exclusively during the larval and nymphal blood meals and (ii) that transcription of bba74 is bracketed by RpoS-independent and -dependent forms of repression that are induced by arthropod- and mammalian host-specific signals, respectively. Although loss of BBA74 does not impair the ability of B. burgdorferi to complete its infectious life cycle, the temporal compartmentalization of this gene's transcription suggests that BBA74 facilitates fitness of the spirochete within a narrow window of its tick phase. A reexamination of the paradigm for reciprocal regulation of ospA and ospC, performed herein, revealed that the heterogeneous expression of OspA and OspC displayed by spirochete populations during the nymphal blood meal results from the intricate sequence of transcriptional and translational changes that ensue as B. burgdorferi transitions between its arthropod vector and mammalian host. PMID- 19218392 TI - Identification and characterization of catabolic para-nitrophenol 4-monooxygenase and para-benzoquinone reductase from Pseudomonas sp. strain WBC-3. AB - Pseudomonas sp. strain WBC-3 utilizes para-nitrophenol (PNP) as a sole source of carbon, nitrogen, and energy. In order to identify the genes involved in this utilization, we cloned and sequenced a 12.7-kb fragment containing a conserved region of NAD(P)H:quinone oxidoreductase genes. Of the products of the 13 open reading frames deduced from this fragment, PnpA shares 24% identity to the large component of a 3-hydroxyphenylacetate hydroxylase from Pseudomonas putida U and PnpB is 58% identical to an NAD(P)H:quinone oxidoreductase from Escherichia coli. Both PnpA and PnpB were purified to homogeneity as His-tagged proteins, and they were considered to be a monomer and a dimer, respectively, as determined by gel filtration. PnpA is a flavin adenine dinucleotide-dependent single-component PNP 4-monooxygenase that converts PNP to para-benzoquinone in the presence of NADPH. PnpB is a flavin mononucleotide-and NADPH-dependent p-benzoquinone reductase that catalyzes the reduction of p-benzoquinone to hydroquinone. PnpB could enhance PnpA activity, and genetic analyses indicated that both pnpA and pnpB play essential roles in PNP mineralization in strain WBC-3. Furthermore, the pnpCDEF gene cluster next to pnpAB shares significant similarities with and has the same organization as a gene cluster responsible for hydroquinone degradation (hapCDEF) in Pseudomonas fluorescens ACB (M. J. Moonen, N. M. Kamerbeek, A. H. Westphal, S. A. Boeren, D. B. Janssen, M. W. Fraaije, and W. J. van Berkel, J. Bacteriol. 190:5190-5198, 2008), suggesting that the genes involved in PNP degradation are physically linked. PMID- 19218393 TI - The Escherichia coli common pilus and the bundle-forming pilus act in concert during the formation of localized adherence by enteropathogenic E. coli. AB - Although the bundle-forming pilus (BFP) of enteropathogenic Escherichia coli (EPEC) mediates microcolony formation on epithelial cells, the adherence of BFP deficient mutants is significantly abrogated, but the mutants are still adherent due to the presence of intimin and possibly other adhesins. In this study we investigated the contribution of the recently described E. coli common pilus (ECP) to the overall adherence properties of EPEC. We found that ECP and BFP structures can be simultaneously observed in the course (between zero time and 7 h during infection) of formation of localized adherence on cultured epithelial cells. These two pilus types colocalized at different levels of the microcolony topology, tethering the adhering bacteria. No evidence of BFP disappearance was found after prolonged infection. When expressed from a plasmid present in nonadherent E. coli HB101, ECP rendered this organism highly adherent at levels comparable to those of HB101 expressing the BFP. Purified ECP bound in a dose dependent manner to epithelial cells, and the binding was blocked with anti-ECP antibodies, confirming that the pili possess adhesin properties. An ECP mutant showed only a modest reduction in adherence to cultured cells due to background expression levels of BFP and intimin. However, isogenic mutants not expressing EspA or BFP were significantly less adherent when the ecpA gene was also deleted. Furthermore, a DeltaespA DeltaecpA double mutant (unable to translocate Tir and to establish intimate adhesion) was at least 10-fold less adherent than the DeltaespA and DeltaecpA single mutants, even in the presence of BFP. A Delta bfp DeltaespA DeltaecpA triple mutant showed the least adherence compared to the wild type and all the isogenic mutant strains tested, suggesting that ECP plays a synergistic role in adherence. Our data indicate that ECP is an accessory factor that, in association with BFP and other adhesins, contributes to the multifactorial complex interaction of EPEC with host epithelial cells. PMID- 19218394 TI - X-Ray crystallographic and mutational studies of fluoroacetate dehalogenase from Burkholderia sp. strain FA1. AB - Fluoroacetate dehalogenase catalyzes the hydrolytic defluorination of fluoroacetate to produce glycolate. The enzyme is unique in that it catalyzes the cleavage of a carbon-fluorine bond of an aliphatic compound: the bond energy of the carbon-fluorine bond is among the highest found in natural products. The enzyme also acts on chloroacetate, although much less efficiently. We here determined the X-ray crystal structure of the enzyme from Burkholderia sp. strain FA1 as the first experimentally determined three-dimensional structure of fluoroacetate dehalogenase. The enzyme belongs to the alpha/beta hydrolase superfamily and exists as a homodimer. Each subunit consists of core and cap domains. The catalytic triad, Asp104-His271-Asp128, of which Asp104 serves as the catalytic nucleophile, was found in the core domain at the domain interface. The active site was composed of Phe34, Asp104, Arg105, Arg108, Asp128, His271, and Phe272 of the core domain and Tyr147, His149, Trp150, and Tyr212 of the cap domain. An electron density peak corresponding to a chloride ion was found in the vicinity of the N(epsilon1) atom of Trp150 and the N(epsilon2) atom of His149, suggesting that these are the halide ion acceptors. Site-directed replacement of each of the active-site residues, except for Trp150, by Ala caused the total loss of the activity toward fluoroacetate and chloroacetate, whereas the replacement of Trp150 caused the loss of the activity only toward fluoroacetate. An interaction between Trp150 and the fluorine atom is probably an absolute requirement for the reduction of the activation energy for the cleavage of the carbon-fluorine bond. PMID- 19218396 TI - Nuclear activity of ROXY1, a glutaredoxin interacting with TGA factors, is required for petal development in Arabidopsis thaliana. AB - Glutaredoxins (GRXs) have thus far been associated mainly with redox-regulated processes participating in stress responses. However, ROXY1, encoding a GRX, has recently been shown to regulate petal primorida initiation and further petal morphogenesis in Arabidopsis thaliana. ROXY1 belongs to a land plant-specific class of GRXs that has a CC-type active site motif, which deviates from ubiquitously occurring CPYC and CGFS GRXs. Expression studies of yellow fluorescent protein-ROXY1 fusion genes driven by the cauliflower mosaic virus 35S promoter reveal a nucleocytoplasmic distribution of ROXY1. We demonstrate that nuclear localization of ROXY1 is indispensable and thus crucial for its activity in flower development. Yeast two-hybrid screens identified TGA transcription factors as interacting proteins, which was confirmed by bimolecular fluorescence complementation experiments showing their nuclear interaction in planta. Overlapping expression patterns of ROXY1 and TGA genes during flower development demonstrate that ROXY1/TGA protein interactions can occur in vivo and support their biological relevance in petal development. Deletion analysis of ROXY1 demonstrates the importance of the C terminus for its functionality and for mediating ROXY1/TGA protein interactions. Phenotypic analysis of the roxy1-2 pan double mutant and an engineered chimeric repressor mutant from PERIANTHIA (PAN), a floral TGA gene, supports a dual role of ROXY1 in petal development. Together, our results show that the ROXY1 protein functions in the nucleus, likely by modifying PAN posttranslationally and thereby regulating its activity in petal primordia initiation. Additionally, ROXY1 affects later petal morphogenesis, probably by modulating other TGA factors that might act redundantly during differentiation of second whorl organs. PMID- 19218397 TI - A novel fatty Acyl-CoA Synthetase is required for pollen development and sporopollenin biosynthesis in Arabidopsis. AB - Acyl-CoA Synthetase (ACOS) genes are related to 4-coumarate:CoA ligase (4CL) but have distinct functions. The Arabidopsis thaliana ACOS5 protein is in clade A of Arabidopsis ACOS proteins, the clade most closely related to 4CL proteins. This clade contains putative nonperoxisomal ACOS enzymes conserved in several angiosperm lineages and in the moss Physcomitrella patens. Although its function is unknown, ACOS5 is preferentially expressed in the flowers of all angiosperms examined. Here, we show that an acos5 mutant produced no pollen in mature anthers and no seeds by self-fertilization and was severely compromised in pollen wall formation apparently lacking sporopollenin or exine. The phenotype was first evident at stage 8 of anther development and correlated with maximum ACOS5 mRNA accumulation in tapetal cells at stages 7 to 8. Green fluorescent protein-ACOS5 fusions showed that ACOS5 is located in the cytoplasm. Recombinant ACOS5 enzyme was active against oleic acid, allowing kinetic constants for ACOS5 substrates to be established. Substrate competition assays indicated broad in vitro preference of the enzyme for medium-chain fatty acids. We propose that ACOS5 encodes an enzyme that participates in a conserved and ancient biochemical pathway required for sporopollenin monomer biosynthesis that may also include the Arabidopsis CYP703A2 and MS2 enzymes. PMID- 19218399 TI - Human pseudogenes of the ABO family show a complex evolutionary dynamics and loss of function. AB - The GT6 glycosyltransferases gene family, that includes the ABO blood group, shows a complex evolution pattern, with multiple events of gain and loss in different mammal species. In humans the ABO gene is considered the sole functional member although the O allele is null and is fixed in certain populations. Here, we analyze the human GT6 pseudogene sequences (Forssman, IGB3, GGTA1, GT6m5, GT6m6, and GT6m7) from an evolutionary perspective, by the study of (i) their diversity levels in populations through the resequencing analysis of European and African individuals; (ii) the interpopulation differentiation, with genotyping data from a survey of populations covering most of human genetic diversity; and (iii) the interespecific divergence, by the comparison of the human and some other primate species sequences. Since pseudogenes are expected to evolve under neutrality, they should show an evolutionary pattern different to that of functional sequences, with higher levels of diversity as well as a ratio of nonsynonymous to synonymous changes close to 1. We describe some departures from these expectations, including selection for inactivation in IGB3, GGTA1, and the interesting case of FS (Forssman) with a probable shift of its initial function in the primate lineage, which put it apart from a pure neutral pseudogene. These results suggest that some of these GT6 human pseudogenes may still be functional and retain some valuable unknown function in humans, in some case even at the protein level. The evolutionary analysis of all members of the GT6 family in humans allows an insight into their functional history, a process likely due to the interaction of the host glycans that they synthesize with pathogens; the past process that can be unraveled through the footprints left by natural selection in the extant genome variation. PMID- 19218398 TI - The E3 ubiquitin ligase SCFTIR1/AFB and membrane sterols play key roles in auxin regulation of endocytosis, recycling, and plasma membrane accumulation of the auxin efflux transporter PIN2 in Arabidopsis thaliana. AB - The PIN family of auxin efflux transporters exhibit polar plasma membrane (PM) localization and play a key role in auxin gradient-mediated developmental processes. Auxin inhibits PIN2 endocytosis and promotes its PM localization. However, the underlying mechanisms remain elusive. Here, we show that the inhibitory effect of auxin on PIN2 endocytosis was impaired in SCF(TIR1/AFB) auxin signaling mutants. Similarly, reducing membrane sterols impaired auxin inhibition of PIN2 endocytosis. Gas chromatography-mass spectrometry analyses indicate that membrane sterols were significantly reduced in SCF(TIR1/AFB) mutants, supporting a link between membrane sterols and auxin signaling in regulating PIN2 endocytosis. We show that auxin promoted PIN2 recycling from endosomes to the PM and increased PIN2 steady state levels in the PM fraction. Furthermore, we show that the positive effect of auxin on PIN2 levels in the PM was impaired by inhibiting membrane sterols or auxin signaling. Consistent with this, the sterol biosynthetic mutant fk-J79 exhibited pronounced defects in primary root elongation and gravitropic response. Our data collectively indicate that, although there are distinct processes involved in endocytic regulation of specific PM-resident proteins, the SCF(TIR1/AFB)-dependent processes are required for auxin regulation of endocytosis, recycling, and PM accumulation of the auxin efflux transporter PIN2 in Arabidopsis thaliana. PMID- 19218400 TI - Comparative analysis of core-fucose-binding lectins from Lens culinaris and Pisum sativum using frontal affinity chromatography. AB - Lens culinaris lectin (LCA) is a useful probe for the detection in serum of a core-fucosylated alpha-fetoprotein, called AFP-L3 fraction, which is a well-known marker for the diagnosis and prognosis of hepatocellular carcinoma. Here we performed a systematic quantitative interaction analysis of LCA and its close homolog, Pisum sativum lectin (PSA), by frontal affinity chromatography with 143 pyridylaminated (PA) glycans including a series of core-fucosylated glycans. Both lectins showed binding affinity to core-fucosylated, mono- and bi-antennary N glycans, but not to their tri- and tetra-antennary forms, indicating that the addition of the GlcNAc residue at the N-acetylglucosaminyltransferase IV position abrogates the binding affinity. However, their specificities are distinguishable: while LCA showed the highest affinity to the core-fucosylated, agalactosylated, bi-antennary N-glycan (K(a)=1.1 x 10(5) M(-1)), PSA showed the highest affinity to the core-fucosylated, trimannosyl structure (K(a)=1.2 x 10(5) M(-1)). Glycan binding specificities of LCA and PSA were also analyzed by glycoconjugate microarray compared to other core-fucose-binding lectins from Aspergillus oryzae (AOL) and Aleuria auratia (AAL). LCA and PSA bound specifically to core fucose, whereas AOL and AAL exhibited broad specificity to fucosylated glycans. These results explain why LCA is appropriate as a specific probe for AFP-L3, which mainly contains a core-fucosylated, biantennary N-glycan, but not its highly branched forms. PMID- 19218401 TI - Time-resolved metabolic footprinting for nonlinear modeling of bacterial substrate utilization. AB - Untargeted profiling of small-molecule metabolites from microbial culture supernatants (metabolic footprinting) has great potential as a phenotyping tool. We used time-resolved metabolic footprinting to compare one Escherichia coli and three Pseudomonas aeruginosa strains growing on complex media and show that considering metabolite changes over the whole course of growth provides much more information than analyses based on data from a single time point. Most strikingly, there was pronounced selectivity in metabolite uptake, even when the bacteria were growing apparently exponentially, with certain groups of metabolites not taken up until others had been entirely depleted from the medium. In addition, metabolite excretion showed some complex patterns. Fitting nonlinear equations (four-parameter sigmoids) to individual metabolite data allowed us to model these changes for metabolite uptake and visualize them by back-projecting the curve-fit parameters onto the original growth curves. These "uptake window" plots clearly demonstrated strain differences, with the uptake of some compounds being reversed in order between different strains. Comparison of an undefined rich medium with a defined complex medium designed to mimic cystic fibrosis sputum showed many differences, both qualitative and quantitative, with a greater proportion of excreted to utilized metabolites in the defined medium. Extending the strain comparison to a more closely related set of isolates showed that it was possible to discriminate two species of the Burkholderia cepacia complex based on uptake dynamics alone. We believe time-resolved metabolic footprinting could be a valuable tool for many questions in bacteriology, including isolate comparisons, phenotyping deletion mutants, and as a functional complement to taxonomic classifications. PMID- 19218402 TI - A novel Dehalobacter species is involved in extensive 4,5,6,7 tetrachlorophthalide dechlorination. AB - The purpose of this study was the enrichment and phylogenetic identification of bacteria that dechlorinate 4,5,6,7-tetrachlorophthalide (commercially designated "fthalide"), an effective fungicide for rice blast disease. Sequential transfer culture of a paddy soil with lactate and fthalide produced a soil-free enrichment culture (designated the "KFL culture") that dechlorinated fthalide by using hydrogen, which is produced from lactate. Phylogenetic analysis based on 16S rRNA genes revealed the dominance of two novel phylotypes of the genus Dehalobacter (FTH1 and FTH2) in the KFL culture. FTH1 and FTH2 disappeared during culture transfer in medium without fthalide and increased in abundance with the dechlorination of fthalide, indicating their growth dependence on the dechlorination of fthalide. Dehalobacter restrictus TEA is their closest relative, with 97.5% and 97.3% 16S rRNA gene similarities to FTH1 and FTH2, respectively. PMID- 19218403 TI - Functional expression of a bacterial xylose isomerase in Saccharomyces cerevisiae. AB - In industrial fermentation processes, the yeast Saccharomyces cerevisiae is commonly used for ethanol production. However, it lacks the ability to ferment pentose sugars like d-xylose and l-arabinose. Heterologous expression of a xylose isomerase (XI) would enable yeast cells to metabolize xylose. However, many attempts to express a prokaryotic XI with high activity in S. cerevisiae have failed so far. We have screened nucleic acid databases for sequences encoding putative XIs and finally were able to clone and successfully express a highly active new kind of XI from the anaerobic bacterium Clostridium phytofermentans in S. cerevisiae. Heterologous expression of this enzyme confers on the yeast cells the ability to metabolize d-xylose and to use it as the sole carbon and energy source. The new enzyme has low sequence similarities to the XIs from Piromyces sp. strain E2 and Thermus thermophilus, which were the only two XIs previously functionally expressed in S. cerevisiae. The activity and kinetic parameters of the new enzyme are comparable to those of the Piromyces XI. Importantly, the new enzyme is far less inhibited by xylitol, which accrues as a side product during xylose fermentation. Furthermore, expression of the gene could be improved by adapting its codon usage to that of the highly expressed glycolytic genes of S. cerevisiae. Expression of the bacterial XI in an industrially employed yeast strain enabled it to grow on xylose and to ferment xylose to ethanol. Thus, our findings provide an excellent starting point for further improvement of xylose fermentation in industrial yeast strains. PMID- 19218405 TI - Development of bacteriocinogenic strains of Saccharomyces cerevisiae heterologously expressing and secreting the leaderless enterocin L50 peptides L50A and L50B from Enterococcus faecium L50. AB - A segregationally stable expression and secretion vector for Saccharomyces cerevisiae, named pYABD01, was constructed by cloning the yeast gene region encoding the mating pheromone alpha-factor 1 secretion signal (MFalpha1(s)) into the S. cerevisiae high-copy-number expression vector pYES2. The structural genes of the two leaderless peptides of enterocin L50 (EntL50A and EntL50B) from Enterococcus faecium L50 were cloned, separately (entL50A or entL50B) and together (entL50AB), into pYABD01 under the control of the galactose-inducible promoter P(GAL1). The generation of recombinant S. cerevisiae strains heterologously expressing and secreting biologically active EntL50A and EntL50B demonstrates the suitability of the MFalpha1(s)-containing vector pYABD01 to direct processing and secretion of these antimicrobial peptides through the S. cerevisiae Sec system. PMID- 19218406 TI - A transferable 20-kilobase multiple drug resistance-conferring R plasmid (pKL0018) from a fish pathogen (Lactococcus garvieae) is highly homologous to a conjugative multiple drug resistance-conferring enterococcal plasmid. AB - Lactococcus garvieae, the causative agent of lactococcosis, has evolved strains that are highly resistant to antibiotics. Here, the 20,034-bp sequence of L. garvieae conjugative plasmid pKL0018 was determined. It contained two ermB genes and one tetS gene and a backbone more than 96% identical to that of pRE25, an Enterococcus faecalis plasmid from dry sausage. PMID- 19218404 TI - Formation of multilayered photosynthetic biofilms in an alkaline thermal spring in Yellowstone National Park, Wyoming. AB - In this study, glass rods suspended at the air-water interface in the runoff channel of Fairy Geyser, Yellowstone National Park, WY, were used as a substratum to promote the development of biofilms that resembled multilayered mat communities in the splash zone at the geyser's source. This approach enabled the establishment of the temporal relationship between the appearance of Cyanobacteria, which ultimately formed the outer green layer, and the development of a red underlayer containing Roseiflexus-like Chloroflexi. This is the first study to define time-dependent successional events involved in the development of differently colored layers within microbial mats associated with many thermal features in Yellowstone National Park. Initial (1-month) biofilms were localized below the air-water interface (60 to 70 degrees C), and the majority of retrieved bacterial sequence types were similar to Synechococcus and Thermus isolates. Biofilms then shifted, becoming established at and above the air-water interface after 3 months. During winter sampling (6 to 8 months), distinct reddish orange microcolonies were observed, consistent with the appearance of Roseiflexus-like sequences and bacteriochlorophyll a pigment signatures. Additionally, populations of Cyanobacteria diversified to include both unicellular and filamentous cell and sequence types. Distinct green and red layers were observed at 13 months. Planctomycetes-like sequences were also retrieved in high abundance from final biofilm layers and winter samples. Finally, biomass associated with geyser vent water contained Roseiflexus-like sequence types, in addition to other high abundance sequence types retrieved from biofilm samples, supporting the idea that geothermal water serves as an inoculum for these habitats. PMID- 19218407 TI - Spontaneous quinolone resistance in the zoonotic serovar of Vibrio vulnificus. AB - This work demonstrates that Vibrio vulnificus biotype 2, serovar E, an eel pathogen able to infect humans, can become resistant to quinolone by specific mutations in gyrA (substitution of isoleucine for serine at position 83) and to some fluoroquinolones by additional mutations in parC (substitution of lysine for serine at position 85). Thus, to avoid the selection of resistant strains that are potentially pathogenic for humans, antibiotics other than quinolones must be used to treat vibriosis on farms. PMID- 19218408 TI - Seasonal dynamics and metagenomic characterization of estuarine viriobenthos assemblages by randomly amplified polymorphic DNA PCR. AB - Direct enumeration and genetic analyses indicate that aquatic sediments harbor abundant and diverse viral communities. Thus far, synecological analysis of estuarine sediment viral diversity over an annual cycle has not been reported. This oversight is due in large part to a lack of molecular genetic approaches for assessing viral diversity within a large collection of environmental samples. Here, randomly amplified polymorphic DNA PCR (RAPD-PCR) was used to examine viral genotypic diversity within Chesapeake Bay sediments. Using a single 10-mer oligonucleotide primer for all samples, RAPD-PCR analysis of sediment viral assemblages yielded unique banding patterns across spatial and temporal scales, with the occurrence of specific bands varying among the sample set. Cluster analysis of RAPD-PCR amplicon banding patterns indicated that sediment viral assemblages changed with season and to a lesser extent with geographic location. Sequence analysis of RAPD-PCR amplicons revealed that 76% of sediment viral sequences were not homologous to any sequence in the GenBank nonredundant protein database. Of the GenBank sequence homologs, the majority belonged to viruses within the Podoviridae (24%) and Myoviridae (22%) viral families, which agrees with the previously observed frequencies of these morphological families in Chesapeake Bay sediments. Furthermore, the majority of the sediment viral sequences homologous to GenBank nonredundant protein sequences were phages or prophages (57%). Hence, RAPD-PCR proved to be a reliable and useful approach for characterization of viral assemblages and the genetic diversity of viruses within aquatic sediments. PMID- 19218409 TI - Variation in bacterial ATP level and proton motive force due to adhesion to a solid surface. AB - Bacterial adhesion to natural and man-made surfaces can be beneficial or detrimental, depending on the system at hand. Of vital importance is how the process of adhesion affects the bacterial metabolic activity. If activity is enhanced, this may help the cells colonize the surface, whereas if activity is reduced, it may inhibit colonization. Here, we report a study demonstrating that adhesion of both Escherichia coli and Bacillus brevis onto a glass surface resulted in enhanced metabolic activity, assessed through ATP measurements. Specifically, ATP levels were found to increase two to five times upon adhesion compared to ATP levels in corresponding planktonic cells. To explain this effect on ATP levels, we propose the hypothesis that bacteria can take advantage of a link between cellular bioenergetics (proton motive force and ATP formation) and the physiochemical charge regulation effect, which occurs as a surface containing ionizable functional groups (e.g., the bacterial cell surface) approaches another surface. As the bacterium approaches the surface, the charge regulation effect causes the charge and pH at the cell surface to vary as a function of separation distance. With negatively charged surfaces, this results in a decrease in pH at the cell surface, which enhances the proton motive force and ATP concentration. Calculations demonstrated that a change in pH across the cell membrane of only 0.2 to 0.5 units is sufficient to achieve the observed ATP increases. Similarly, the hypothesis indicates that positively charged surfaces will decrease metabolic activity, and results from studies of positively charged surfaces support this finding. PMID- 19218410 TI - New electropositive filter for concentrating enteroviruses and noroviruses from large volumes of water. AB - The U.S. Environmental Protection Agency's information collection rule requires the use of 1MDS electropositive filters for concentrating enteric viruses from water, but unfortunately, these filters are not cost-effective for routine viral monitoring. In this study, an inexpensive electropositive cartridge filter, the NanoCeram filter, was evaluated for its ability to concentrate enteroviruses and noroviruses from large volumes of water. Seeded viruses were concentrated using the adsorption-elution procedure. The mean percent retention of seeded polioviruses by NanoCeram filters was 84%. To optimize the elution procedure, six protocols, each comprising two successive elutions with various lengths of filter immersion, were evaluated. The highest virus recovery (77%) was obtained by immersing the filters in beef extract for 1 minute during the first elution and for 15 min during the second elution. The recovery efficiencies of poliovirus, coxsackievirus B5, and echovirus 7 from 100-liter samples of seeded tap water were 54%, 27%, and 32%, respectively. There was no significant difference in virus recovery from tap water with a pH range of 6 to 9.5 and a water flow rate range of 5.5 liters/min to 20 liters/min. Finally, poliovirus and Norwalk virus recoveries by NanoCeram filters were compared to those by 1MDS filters, using tap water and Ohio River water. Poliovirus and Norwalk virus recoveries by NanoCeram filters from tap and river water were similar to or higher than those by the 1MDS filters. These data suggest that NanoCeram filters can be used as an inexpensive alternative to 1MDS filters for routine viral monitoring of water. PMID- 19218412 TI - Isolation and characterization of metalloproteases with a novel domain structure by construction and screening of metagenomic libraries. AB - Small-insert metagenomic libraries from four samples were constructed by a topoisomerase-based and a T4 DNA ligase-based approach. Direct comparison of both approaches revealed that application of the topoisomerase-based method resulted in a higher number of insert-containing clones per microg of environmental DNA used for cloning and a larger average insert size. Subsequently, the constructed libraries were partially screened for the presence of genes conferring proteolytic activity. The function-driven screen was based on the ability of the library-containing Escherichia coli clones to form halos on skim milk-containing agar plates. The screening of 80,000 E. coli clones yielded four positive clones. Two of the plasmids (pTW2 and pTW3) recovered from positive clones conferred strong proteolytic activity and were studied further. Analysis of the entire insert sequences of pTW2 (28,113 bp) and pTW3 (19,956 bp) suggested that the DNA fragments were derived from members of the genus Xanthomonas. Each of the plasmids harbored one gene (2,589 bp) encoding a metalloprotease (mprA, pTW2; mprB, pTW3). Sequence and biochemical analyses revealed that MprA and MprB are similar extracellular proteases belonging to the M4 family of metallopeptidases (thermolysin-like family). Both enzymes possessed a unique modular structure and consisted of four regions: the signal sequence, the N-terminal proregion, the protease region, and the C-terminal extension. The architecture of the latter region, which was characterized by the presence of two prepeptidase C-terminal domains and one proprotein convertase P domain, is novel for bacterial metalloproteases. Studies with derivatives of MprA and MprB revealed that the C terminal extension is not essential for protease activity. The optimum pH and temperature of both proteases were 8.0 and 65 degrees C, respectively, when casein was used as substrate. PMID- 19218411 TI - Production of the phytohormone indole-3-acetic acid by estuarine species of the genus Vibrio. AB - Strains of Vibrio spp. isolated from roots of the estuarine grasses Spartina alterniflora and Juncus roemerianus produce the phytohormone indole-3-acetic acid (IAA). The colorimetric Salkowski assay was used for initial screening of IAA production. Gas chromatography-mass spectroscopy (GC-MS) was then employed to confirm and quantify IAA production. The accuracy of IAA quantification by the Salkowski assay was examined by comparison to GC-MS assay values. Indole-3 acetamide, an intermediate in IAA biosynthesis by the indole-3-acetamide pathway, was also identified by GC-MS. Multilocus sequence typing of concatenated 16S rRNA, recA, and rpoA genes was used for phylogenetic analysis of environmental isolates within the genus Vibrio. Eight Vibrio type strains and five additional species-level clades containing a total of 16 environmental isolates and representing five presumptive new species were identified as IAA-producing Vibrio species. Six additional environmental isolates similar to four of the Vibrio type strains were also IAA producers. To our knowledge, this is the first report of IAA production by species of the genus Vibrio or by bacteria isolated from an estuarine environment. PMID- 19218413 TI - Oenococcus oeni genome plasticity is associated with fitness. AB - Oenococcus oeni strains are well-known for their considerable phenotypic variations in terms of tolerance to harsh wine conditions and malolactic activity. Genomic subtractive hybridization (SH) between two isolates with differing enological potentials was used to elucidate the genetic bases of this intraspecies diversity and identify novel genes involved in adaptation to wine. SH revealed 182 tester-specific fragments corresponding to 126 open reading frames (ORFs). A large proportion of the chromosome-related ORFs resembled genes involved in carbohydrate transport and metabolism, cell wall/membrane/envelope biogenesis, and replication, recombination, and repair. Six regions of genomic plasticity were identified, and their analysis suggested that both limited recombination and insertion/deletion events contributed to the vast genomic diversity observed in O. oeni. The association of selected sequences with adaptation to wine was further assessed by screening a large collection of strains using PCR. No sequences were found to be specific to highly performing (HP) strains alone. However, there was a statistically significant positive association between HP strains and the presence of eight gene sequences located on regions 2, 4, and 5. Gene expression patterns were significantly modified in HP strains, following exposure to one or more of the common stresses in wines. Regions 2 and 5 showed no traces of mobile elements and had normal GC content. In contrast, region 4 had the typical hallmarks of horizontal transfer, suggesting that the strategy of acquiring genes from other bacteria enhances the fitness of O. oeni strains. PMID- 19218414 TI - Characterization of regulatory pathways in Xylella fastidiosa: genes and phenotypes controlled by gacA. AB - The xylem-limited, insect-transmitted bacterium Xylella fastidiosa causes Pierce's disease in grapes through cell aggregation and vascular clogging. GacA controls various physiological processes and pathogenicity factors in many gram negative bacteria, including biofilm formation in Pseudomonas syringae pv. tomato DC3000. Cloned gacA of X. fastidiosa was found to restore the hypersensitive response and pathogenicity in gacA mutants of P. syringae pv. tomato DC3000 and Erwinia amylovora. A gacA mutant of X. fastidiosa (DAC1984) had significantly reduced abilities to adhere to a glass surface, form biofilm, and incite disease symptoms on grapevines, compared with the parent (A05). cDNA microarray analysis identified 7 genes that were positively regulated by GacA, including xadA and hsf, predicted to encode outer membrane adhesion proteins, and 20 negatively regulated genes, including gumC and an antibacterial polypeptide toxin gene, cvaC. These results suggest that GacA of X. fastidiosa regulates many factors, which contribute to attachment and biofilm formation, as well as some physiological processes that may enhance the adaptation and tolerance of X. fastidiosa to environmental stresses and the competition within the host xylem. PMID- 19218415 TI - Filamentous bacterium Eikelboom type 0092 in activated sludge plants in Australia is a member of the phylum Chloroflexi. AB - Molecular data show that the filamentous bacterium Eikelboom type 0092, frequently seen in Australian activated sludge plants, is a member of the phylum Chloroflexi. Fluorescence in situ hybridization (FISH) probes designed against cloned 16S rRNA sequences from a full-scale enhanced biological phosphate removal activated sludge plant community, where this was a dominant filament morphotype, suggest that it can exist as two variants, differing in their trichome diameter. When applied to samples from several treatment plants in eastern Australia, each FISH probe targeted only the type 0092 filament morphotype against which it was designed. The patterns of FISH signals generated with both were consistent with the ribosomes not being evenly distributed but arranged as intracellular aggregates. The FISH survey data showed that these two variants appeared together in most but not all of the plants examined. None stained positively for intracellular presence of either poly-beta-hydroxyalkanoates or polyphosphate. PMID- 19218416 TI - Rhizobia from Lanzarote, the Canary Islands, that nodulate Phaseolus vulgaris have characteristics in common with Sinorhizobium meliloti isolates from mainland Spain. AB - The stable, low-molecular-weight (LMW) RNA fractions of several rhizobial isolates of Phaseolus vulgaris grown in the soil of Lanzarote, an island of the Canary Islands, were identical to a less-common pattern found within Sinorhizobium meliloti (assigned to group II) obtained from nodules of alfalfa and alfalfa-related legumes grown in northern Spain. The P. vulgaris isolates and the group II LMW RNA S. meliloti isolates also were distinguishable in that both had two conserved inserts of 20 and 46 bp in the 16S-23S internal transcribed spacer region that were not present in other strains of S. meliloti. The isolates from P. vulgaris nodulated bean but not Medicago sativa, while those recovered from Medicago, Melilotus, and Trigonella spp. nodulated both host legumes. The bean isolates also were distinguished from those of Medicago, Melilotus, and Trigonella spp. by nodC sequence analysis. The nodC sequences of the bean isolates were most similar to those reported for S. meliloti bv. mediterranense and Sinorhizobium fredii bv. mediterranense (GenBank accession numbers DQ333891 and AF217267, respectively). None of the evidence placed the bean isolates from Lanzarote in the genus Rhizobium, which perhaps is inconsistent with seed-borne transmission of Rhizobium etli from the Americas to the Canaries as an explanation for the presence of bean-nodulating rhizobia in soils of Lanzarote. PMID- 19218418 TI - Elimination of Escherichia coli O157:H7 from Alfalfa seeds through a combination of high hydrostatic pressure and mild heat. AB - Escherichia coli O157:H7 has been associated with contaminated seed sprout outbreaks. The majority of these outbreaks have been traced to sprout seeds contaminated with low levels of pathogens. Sanitizing sprout seeds presents a unique challenge in the arena of produce safety in that even a low residual pathogen population remaining on contaminated seed after treatments appears capable of growing to very high levels during sprouting. In this study, the effectiveness of high-pressure treatment in combination with low and elevated temperatures was assessed for its ability to eliminate E. coli O157:H7 on artificially contaminated alfalfa seeds. Inoculated seed samples were treated at 600 MPa for 2 min at 4, 20, 25, 30, 35, 40, 45, and 50 degrees C. The pressure sensitivity of the pathogenic bacteria was strongly dependent on the treatment temperature. At 40 degrees C, the process was adequate in eliminating a 5-log unit population on the seeds with no adverse effect on seed viability. Three treatments carried out at reduced pressure levels and/or extended treatment time, 550 MPa for 2 min at 40 degrees C, 300 MPa for 2 min at 50 degrees C, and 400 MPa for 5 min at 45 degrees C, were equally lethal to the pathogen. When all three treatments were compared in terms of their impact on seed viability, the process of 550 MPa for 2 min at 40 degrees C was the most desirable, achieving final germination percentages and sprout sizes statistically similar to those of control untreated seeds (P > 0.05). PMID- 19218417 TI - Effect of biowaste sludge maturation on the diversity of thermophilic bacteria and archaea in an anaerobic reactor. AB - Prokaryotic diversity was investigated near the inlet and outlet of a plug-flow reactor. After analyzing 800 clones, 50 bacterial and 3 archaeal phylogenetic groups were defined. Clostridia (>92%) dominated among bacteria and Methanoculleus (>90%) among archaea. Significant changes in pH and volatile fatty acids did not invoke a major shift in the phylogenetic groups. We suggest that the environmental filter imposed by the saline conditions (20 g liter(-1)) selected a stable community of halotolerant and halophilic prokaryotes. PMID- 19218419 TI - Hierarchical oligonucleotide primer extension as a time- and cost-effective approach for quantitative determination of Bifidobacterium spp. in infant feces. AB - The Bifidobacterium spp. present in 10 infant fecal samples (4 from infants with eczema and 6 from healthy infants) were quantified with both hierarchical oligonucleotide primer extension (HOPE) and fluorescence in situ hybridization flow cytometry. The relative abundances of Bifidobacterium longum and B. catenulatum with respect to the total bifidobacteria had a poor correlation (rho, <0.600; P value, >0.208), presumably due to differences in primer specificity and the level of hybridization stringency of both methods. In contrast, the relative abundances of organisms of the genus Bifidobacterium against the total amplified 16S rRNA genes and those of B. adolescentis, B. bifidum, and B. breve against the genus Bifidobacterium exhibited a good statistical correlation (rho, >0.783; P value, <0.066). This good comparability supports HOPE as a method to achieve high throughput quantitative determination of bacterial targets in a time- and cost effective manner. PMID- 19218420 TI - Ecophysiology of "Halarsenatibacter silvermanii" strain SLAS-1T, gen. nov., sp. nov., a facultative chemoautotrophic arsenate respirer from salt-saturated Searles Lake, California. AB - Searles Lake occupies a closed basin harboring salt-saturated, alkaline brines that have exceptionally high concentrations of arsenic oxyanions. Strain SLAS 1(T) was previously isolated from Searles Lake (R. S. Oremland, T. R. Kulp, J. Switzer Blum, S. E. Hoeft, S. Baesman, L. G. Miller, and J. F. Stolz, Science 308:1305-1308, 2005). We now describe this extremophile with regard to its substrate affinities, its unusual mode of motility, sequenced arrABD gene cluster, cell envelope lipids, and its phylogenetic alignment within the order Halanaerobacteriales, assigning it the name "Halarsenatibacter silvermanii" strain SLAS-1(T). We also report on the substrate dynamics of an anaerobic enrichment culture obtained from Searles Lake that grows under conditions of salt saturation and whose members include a novel sulfate reducer of the order Desulfovibriales, the archaeon Halorhabdus utahensis, as well as a close homolog of strain SLAS-1(T). PMID- 19218421 TI - Community composition and density of methanogens in the foregut of the Tammar wallaby (Macropus eugenii). AB - The composition of the methanogenic archaeal community in the foregut contents of Tammar wallabies (Macropus eugenii) was studied using 16S rRNA and methyl coenzyme reductase subunit A (mcrA) gene clone libraries. Methanogens belonging to the Methanobacteriales and a well-supported cluster of uncultivated archaeon sequences previously observed in the ovine and bovine rumens were found. Methanogen densities ranged from 7.0 x 10(5) and 3.9 x 10(6) cells per gram of wet weight. PMID- 19218422 TI - p21-activated kinases Cla4 and Ste20 regulate vacuole inheritance in Saccharomyces cerevisiae. AB - Each time Saccharomyces cerevisiae cells divide they ensure that both the mother and daughter cell inherit a vacuole by actively transporting a portion of the vacuole into the bud. As the mother cell begins budding, a tubular and vesicular segregation structure forms that is transported into the bud by the myosin V motor Myo2, which is bound to the vacuole-specific myosin receptor, Vac17 (41, 59, 70, 79). Upon arriving in the bud the segregation structure is resolved to found the daughter vacuole. The mechanism that regulates segregation structure resolution in a spatially dependent manner is unknown. In addition to resolving the segregation structure, Vac17 is degraded specifically in the bud to provide directionality to vacuole inheritance. It has been proposed that bud-specific degradation of Vac17 is promoted by proteins localized to or activated solely in the bud (77). The p21-activated kinases (PAKs) Cla4 and Ste20 are localized to and activated in the bud. Here we report that Cla4 is localized to the segregation structure just prior to segregation structure resolution, and cells lacking PAK function fail to resolve the segregation structure. Overexpression of either Cla4 or Ste20 inhibited vacuole inheritance and this inhibition was suppressed by the expression of nondegradable VAC17. Finally, PAK activity was required for Vac17 degradation in late M phase and CLA4 overexpression promoted Vac17 degradation. We propose that Cla4 and Ste20 are bud-specific proteins that play roles in both segregation structure resolution and the degradation of Vac17. PMID- 19218423 TI - Efficient gene replacements in Toxoplasma gondii strains deficient for nonhomologous end joining. AB - A high frequency of nonhomologous recombination has hampered gene targeting approaches in the model apicomplexan parasite Toxoplasma gondii. To address whether the nonhomologous end-joining (NHEJ) DNA repair pathway could be disrupted in this obligate intracellular parasite, putative KU proteins were identified and a predicted KU80 gene was deleted. The efficiency of gene targeting via double-crossover homologous recombination at several genetic loci was found to be greater than 97% of the total transformants in KU80 knockouts. Gene replacement efficiency was markedly increased (300- to 400-fold) in KU80 knockouts compared to wild-type strains. Target DNA flanks of only approximately 500 bp were found to be sufficient for efficient gene replacements in KU80 knockouts. KU80 knockouts stably retained a normal growth rate in vitro and the high virulence phenotype of type I strains but exhibited an increased sensitivity to double-strand DNA breaks induced by treatment with phleomycin or gamma irradiation. Collectively, these results revealed that a significant KU-dependent NHEJ DNA repair pathway is present in Toxoplasma gondii. Integration essentially occurs only at the homologous targeted sites in the KU80 knockout background, making this genetic background an efficient host for gene targeting to speed postgenome functional analysis and genetic dissection of parasite biology. PMID- 19218424 TI - Degradation of Saccharomyces cerevisiae transcription factor Gcn4 requires a C terminal nuclear localization signal in the cyclin Pcl5. AB - Pcl5 is a Saccharomyces cerevisiae cyclin that directs the phosphorylation of the general amino acid control transcriptional activator Gcn4 by the cyclin-dependent kinase (CDK) Pho85. Phosphorylation of Gcn4 by Pho85/Pcl5 initiates its degradation via the ubiquitin/proteasome system and is regulated by the availability of amino acids. In this study, we show that Pcl5 is a nuclear protein and that artificial dislocation of Pcl5 into the cytoplasm prevents the degradation of Gcn4. Nuclear localization of Pcl5 depends on the beta-importin Kap95 and does not require Pho85, Gcn4, or the CDK inhibitor Pho81. Pcl5 nuclear import is independent on the availability of amino acids and is mediated by sequences in its C-terminal domain. The nuclear localization signal is distinct from other functional domains of Pcl5. This is corroborated by a C-terminally truncated Pcl5 variant, which carries the N-terminal nuclear domain of Pho80. This hybrid is still able to fulfill Pcl5 function, whereas Pho80, which is another Pho85 interacting cyclin, does not mediate Gcn4 degradation. PMID- 19218425 TI - Hog1 mitogen-activated protein kinase (MAPK) interrupts signal transduction between the Kss1 MAPK and the Tec1 transcription factor to maintain pathway specificity. AB - In Saccharomyces cerevisiae, the mating, filamentous growth (FG), and high osmolarity glycerol (HOG) mitogen-activated protein kinase (MAPK) signaling pathways share components and yet mediate distinct responses to different extracellular signals. Cross talk is suppressed between the mating and FG pathways because mating signaling induces the destruction of the FG transcription factor Tec1. We show here that HOG pathway activation results in phosphorylation of the FG MAPK, Kss1, and the MAPKK, Ste7. However, FG transcription is not activated because HOG signaling prevents the activation of Tec1. In contrast to the mating pathway, we find that the mechanism involves the inhibition of DNA binding by Tec1 rather than its destruction. We also find that nuclear accumulation of Tec1 is not affected by HOG signaling. Inhibition by Hog1 is apparently indirect since it does not require any of the consensus S/TP MAPK phosphorylation sites on Tec1, its DNA-binding partner Ste12, or the associated regulators Dig1 or Dig2. It also does not require the consensus MAPK sites of the Ste11 activator Ste50, in contrast to a recent proposal for a role for negative feedback in specificity. Our results demonstrate that HOG signaling interrupts the FG pathway signal transduction between the phosphorylation of Kss1 and the activation of DNA binding by Tec1. PMID- 19218426 TI - Tagging of endogenous genes in a Toxoplasma gondii strain lacking Ku80. AB - As with other organisms with a completed genome sequence, opportunities for performing large-scale studies, such as expression and localization, on Toxoplasma gondii are now much more feasible. We present a system for tagging genes endogenously with yellow fluorescent protein (YFP) in a Deltaku80 strain. Ku80 is involved in DNA strand repair and nonhomologous DNA end joining; previous studies in other organisms have shown that in its absence, random integration is eliminated, allowing the insertion of constructs with homologous sequences into the proper loci. We generated a vector consisting of YFP and a dihydrofolate reductase-thymidylate synthase selectable marker. The YFP is preceded by a ligation-independent cloning (LIC) cassette, which allows the insertion of PCR products containing complementary LIC sequences. We demonstrated that the Deltaku80 strain is more effective and efficient in integrating the YFP-tagged constructs into the correct locus than wild-type strain RH. We then selected several hypothetical proteins that were identified by a proteomic screen of excreted-secreted antigens and that displayed microarray expression profiles similar to known micronemal proteins, with the thought that these could potentially be new proteins with roles in cell invasion. We localized these hypothetical proteins by YFP fluorescence and showed expression by immunoblotting. Our findings demonstrate that the combination of the Deltaku80 strain and the pYFP.LIC constructs reduces both the time and cost required to determine localization of a new gene of interest. This should allow the opportunity for performing larger-scale studies of novel T. gondii genes. PMID- 19218427 TI - Persistent elimination of ErbB-2/HER2-overexpressing tumors using combinations of monoclonal antibodies: relevance of receptor endocytosis. AB - Monoclonal antibodies (mAbs) to ErbB-2/HER2 or to its sibling, the epidermal growth factor receptor (EGFR), prolong survival of cancer patients, especially when combined with cytotoxic therapies. However, low effectiveness of therapeutic mAbs and the evolution of patient resistance call for improvements. Here we test in animals pairs of anti-ErbB-2 mAbs and report that pairs comprising an antibody reactive with the dimerization site of ErbB-2 and an antibody recognizing another distinct epitope better inhibit ErbB-2-overexpressing tumors than other pairs or the respective individual mAbs. Because the superiority of antibody combinations extends to tumor cell cultures, we assume that nonimmunological mechanisms contribute to mAb synergy. One potential mechanism, namely the ability of mAb combinations to instigate ErbB-2 endocytosis, is demonstrated. Translation of these lessons to clinical applications may enhance patient response and delay acquisition of resistance. PMID- 19218428 TI - A rosette-type, self-renewing human ES cell-derived neural stem cell with potential for in vitro instruction and synaptic integration. AB - An intriguing question in human embryonic stem cell (hESC) biology is whether these pluripotent cells can give rise to stably expandable somatic stem cells, which are still amenable to extrinsic fate instruction. Here, we present a pure population of long-term self-renewing rosette-type hESC-derived neural stem cells (lt-hESNSCs), which exhibit extensive self-renewal, clonogenicity, and stable neurogenesis. Although lt-hESNSCs show a restricted expression of regional transcription factors, they retain responsiveness to instructive cues promoting the induction of distinct subpopulations, such as ventral midbrain and spinal cord fates. Using lt-hESNSCs as a donor source for neural transplantation, we provide direct evidence that hESC-derived neurons can establish synaptic connectivity with the mammalian nervous system. Combining long-term stability, maintenance of rosette-properties and phenotypic plasticity, lt-hESNSCs may serve as useful tool to study mechanisms of human NSC self-renewal, lineage segregation, and functional in vivo integration. PMID- 19218429 TI - CXCL14 is an autocrine growth factor for fibroblasts and acts as a multi-modal stimulator of prostate tumor growth. AB - This study explored the role of secreted fibroblast-derived factors in prostate cancer growth. Analyses of matched normal and tumor tissue revealed up-regulation of CXCL14 in cancer-associated fibroblasts of a majority of prostate cancer. Fibroblasts over-expressing CXCL14 promoted the growth of prostate cancer xenografts, and increased tumor angiogenesis and macrophage infiltration. Mechanistic studies demonstrated that autocrine CXCL14-stimulation of fibroblasts stimulate migration and ERK-dependent proliferation of fibroblasts. CXCL14 stimulation of monocyte migration was also demonstrated. Furthermore, CXCL14 producing fibroblasts, but not recombinant CXCL14, enhanced in vitro proliferation and migration of prostate cancer cells and in vivo angiogenesis. These studies thus identify CXCL14 as a novel autocrine stimulator of fibroblast growth and migration, with multi-modal tumor-stimulatory activities. In more general terms, our findings suggest autocrine stimulation of fibroblasts as a previously unrecognized mechanism for chemokine-mediated stimulation of tumor growth, and suggest a novel mechanism whereby cancer-associated fibroblasts achieve their pro-tumorigenic phenotype. PMID- 19218430 TI - Dysregulated gene expression networks in human acute myelogenous leukemia stem cells. AB - We performed the first genome-wide expression analysis directly comparing the expression profile of highly enriched normal human hematopoietic stem cells (HSC) and leukemic stem cells (LSC) from patients with acute myeloid leukemia (AML). Comparing the expression signature of normal HSC to that of LSC, we identified 3,005 differentially expressed genes. Using 2 independent analyses, we identified multiple pathways that are aberrantly regulated in leukemic stem cells compared with normal HSC. Several pathways, including Wnt signaling, MAP Kinase signaling, and Adherens Junction, are well known for their role in cancer development and stem cell biology. Other pathways have not been previously implicated in the regulation of cancer stem cell functions, including Ribosome and T Cell Receptor Signaling pathway. This study demonstrates that combining global gene expression analysis with detailed annotated pathway resources applied to highly enriched normal and malignant stem cell populations, can yield an understanding of the critical pathways regulating cancer stem cells. PMID- 19218431 TI - Specific synthetic lethal killing of RAD54B-deficient human colorectal cancer cells by FEN1 silencing. AB - Mutations that cause chromosome instability (CIN) in cancer cells produce "sublethal" deficiencies in an essential process (chromosome segregation) and, therefore, may represent a major untapped resource that could be exploited for therapeutic benefit in the treatment of cancer. If second-site unlinked genes can be identified, that when knocked down, cause a synthetic lethal (SL) phenotype in combination with a somatic mutation in a CIN gene, novel candidate therapeutic targets will be identified. To test this idea, we took a cross species SL candidate gene approach by recapitulating a SL interaction observed between rad54 and rad27 mutations in yeast, via knockdown of the highly sequence- and functionally-related proteins RAD54B and FEN1 in a cancer cell line. We show that knockdown of RAD54B, a gene known to be somatically mutated in cancer, causes CIN in mammalian cells. Using high-content microscopy techniques, we demonstrate that RAD54B-deficient human colorectal cancer cells are sensitive to SL killing by reduced FEN1 expression, while isogenic RAD54B proficient cells are not. This conserved SL interaction suggests that extrapolating SL interactions observed in model organisms for homologous genes mutated in human cancers will aid in the identification of novel therapeutic targets for specific killing of cancerous cells exhibiting CIN. PMID- 19218432 TI - Dual involvement of G-substrate in motor learning revealed by gene deletion. AB - In this study, we generated mice lacking the gene for G-substrate, a specific substrate for cGMP-dependent protein kinase uniquely located in cerebellar Purkinje cells, and explored their specific functional deficits. G-substrate deficient Purkinje cells in slices obtained at postnatal weeks (PWs) 10-15 maintained electrophysiological properties essentially similar to those from WT littermates. Conjunction of parallel fiber stimulation and depolarizing pulses induced long-term depression (LTD) normally. At younger ages, however, LTD attenuated temporarily at PW6 and recovered thereafter. In parallel with LTD, short-term (1 h) adaptation of optokinetic eye movement response (OKR) temporarily diminished at PW6. Young adult G-substrate knockout mice tested at PW12 exhibited no significant differences from their WT littermates in terms of brain structure, general behavior, locomotor behavior on a rotor rod or treadmill, eyeblink conditioning, dynamic characteristics of OKR, or short-term OKR adaptation. One unique change detected was a modest but significant attenuation in the long-term (5 days) adaptation of OKR. The present results support the concept that LTD is causal to short-term adaptation and reveal the dual functional involvement of G-substrate in neuronal mechanisms of the cerebellum for both short-term and long-term adaptation. PMID- 19218433 TI - Murine epidermal Langerhans cells and langerin-expressing dermal dendritic cells are unrelated and exhibit distinct functions. AB - A new langerin(+) DC subset has recently been identified in murine dermis (langerin(+) dDC), but the lineage and functional relationships between these cells and langerin(+) epidermal Langerhans cells (LC) are incompletely characterized. Selective expression of the cell adhesion molecule EpCAM by LC allowed viable LC to be easily distinguished from langerin(+) dDC in skin and lymphoid tissue and ex vivo as well. Differential expression of EpCAM and langerin revealed the presence of at least 3 distinct skin DC subsets. We determined that LC and langerin(+) dDC exhibit different migratory capabilities in vitro and repopulate distinct anatomic compartments in skin at different rates after conditional depletion in vivo. Langerin(+) dDC, in contrast to LC, did not require TGFbeta1 for development. Carefully timed gene gun immunization studies designed to take advantage of the distinct repopulation kinetics of langerin(+) dDC and LC revealed that langerin(+) dDC were required for optimal production of beta-galactosidase-specific IgG2a/c and IgG2b in the acute phase. In contrast, immunization via LC-deficient skin resulted in persistent and strikingly reduced IgG1 and enhanced IgG2a Ab production. Our data support the concepts that LC and langerin(+) dDC represent distinct DC subsets that have specialized functions and that LC are important immunoregulatory cells. The presence of at least 3 functionally distinct skin DC subsets may have particular relevance for vaccines that are administered epicutaneously. PMID- 19218434 TI - Hedgehog signal transduction by Smoothened: pharmacologic evidence for a 2-step activation process. AB - The Hedgehog (Hh) signaling pathway controls growth, cell fate decisions, and morphogenesis during development. Damage to Hh transduction machinery can lead to birth defects and cancer. The transmembrane protein Smoothened (Smo) relays the Hh signal and is an important drug target in cancer. Smo enrichment in primary cilia is thought to drive activation of target genes. Using small-molecule agonists and antagonists to dissect Smo function, we find that Smo enrichment in cilia is not sufficient for signaling and a distinct second step is required for full activation. This 2-step mechanism--localization followed by activation--has direct implications for the design and use of anticancer therapeutics targeted against Smo. PMID- 19218435 TI - The minimum information principle and its application to neural code analysis. AB - The study of complex information processing systems requires appropriate theoretical tools to help unravel their underlying design principles. Information theory is one such tool, and has been utilized extensively in the study of the neural code. Although much progress has been made in information theoretic methodology, there is still no satisfying answer to the question: "What is the information that a given property of the neural population activity (e.g., the responses of single cells within the population) carries about a set of stimuli?" Here, we answer such questions via the minimum mutual information (MinMI) principle. We quantify the information in any statistical property of the neural response by considering all hypothetical neuronal populations that have the given property and finding the one that contains the minimum information about the stimuli. All systems with higher information values necessarily contain additional information processing mechanisms and, thus, the minimum captures the information related to the given property alone. MinMI may be used to measure information in properties of the neural response, such as that conveyed by responses of small subsets of cells (e.g., singles or pairs) in a large population and cooperative effects between subunits in networks. We show how the framework can be used to study neural coding in large populations and to reveal properties that are not discovered by other information theoretic methods. PMID- 19218436 TI - Oxidant regulated inter-subunit disulfide bond formation between ASIC1a subunits. AB - The acid-sensing ion channel-1a (ASIC1a) is composed of 3 subunits and is activated by a decrease in extracellular pH. It plays an important role in diseases associated with a reduced pH and production of oxidants. Previous work showed that oxidants reduce ASIC1a currents. However, the effects on channel structure and composition are unknown. We found that ASIC1a formed inter-subunit disulfide bonds and the oxidant H(2)O(2) increased this link between subunits. Cys-495 in the ASIC1a C terminus was particularly important for inter-subunit disulfide bond formation, although other C-terminal cysteines contributed. Inter subunit disulfide bonds also produced some ASIC1a complexes larger than trimers. Inter-subunit disulfide bond formation reduced the proportion of ASIC1a located on the cell surface and contributed to the H(2)O(2)-induced decrease in H(+) gated current. These results indicate that channel function is controlled by disulfide bond formation between intracellular residues on distinct ASIC1a subunits. They also suggest a mechanism by which the redox state can dynamically regulate membrane protein activity by forming intracellular bridges. PMID- 19218437 TI - Donor Toll-like receptor 4 contributes to ischemia and reperfusion injury following human kidney transplantation. AB - While studies in animal models have linked Toll-like receptor (TLR) 4 signaling to kidney injury induced by ischemia and reperfusion, the relevance of TLR4 activation to allograft injury in human kidney transplants is unknown. Here we show that TLR4 is constitutively expressed within all donor kidneys but is significantly higher in deceased-, compared with living-donor organs. Tubules from deceased- but not living-donor kidneys also stained positively for high mobility group box-1 (HMGB1), a known endogenous TLR4 ligand. In vitro stimulation of human tubular cells with HMGB1, in a TLR4-dependent system, confirmed that HMGB1 can stimulate proinflammatory responses through TLR4. To assess the functional significance of TLR4 in human kidney transplantation, we determined whether TLR4 mutations that confer diminished affinity for HMGB1 influence intragraft gene-expression profiles and immediate graft function. Compared with kidneys expressing WT alleles, kidneys with a TLR4 loss-of-function allele contained less TNFalpha, MCP-1, and more heme oxygenase 1 (HO-1), and exhibited a higher rate of immediate graft function. These results represent previously undetected evidence that donor TLR4 contributes to graft inflammation and sterile injury following cold preservation and transplantation in humans. Targeting TLR4 signaling may have value in preventing or treating postischemic acute kidney injury after transplantation. PMID- 19218438 TI - The B-type lamin is required for somatic repression of testis-specific gene clusters. AB - Large clusters of coexpressed tissue-specific genes are abundant on chromosomes of diverse species. The genes coordinately misexpressed in diverse diseases are also found in similar clusters, suggesting that evolutionarily conserved mechanisms regulate expression of large multigenic regions both in normal development and in its pathological disruptions. Studies on individual loci suggest that silent clusters of coregulated genes are embedded in repressed chromatin domains, often localized to the nuclear periphery. To test this model at the genome-wide scale, we studied transcriptional regulation of large testis specific gene clusters in somatic tissues of Drosophila. These gene clusters showed a drastic paucity of known expressed transgene insertions, indicating that they indeed are embedded in repressed chromatin. Bioinformatics analysis suggested the major role for the B-type lamin, LamDm(o), in repression of large testis-specific gene clusters, showing that in somatic cells as many as three quarters of these clusters interact with LamDm(o). Ablation of LamDm(o) by using mutants and RNAi led to detachment of testis-specific clusters from nuclear envelope and to their selective transcriptional up-regulation in somatic cells, thus providing the first direct evidence for involvement of the B-type lamin in tissue-specific gene repression. Finally, we found that transcriptional activation of the lamina-bound testis-specific gene cluster in male germ line is coupled with its translocation away from the nuclear envelope. Our studies, which directly link nuclear architecture with coordinated regulation of tissue-specific genes, advance understanding of the mechanisms underlying both normal cell differentiation and developmental disorders caused by lesions in the B-type lamins and interacting proteins. PMID- 19218440 TI - Two-slit diffraction with highly charged particles: Niels Bohr's consistency argument that the electromagnetic field must be quantized. AB - We analyze Niels Bohr's proposed two-slit interference experiment with highly charged particles which argues that the consistency of elementary quantum mechanics requires that the electromagnetic field must be quantized. In the experiment a particle's path through the slits is determined by measuring the Coulomb field that it produces at large distances; under these conditions the interference pattern must be suppressed. The key is that, as the particle's trajectory is bent in diffraction by the slits, it must radiate and the radiation must carry away phase information. Thus, the radiation field must be a quantized dynamical degree of freedom. However, if one similarly tries to determine the path of a massive particle through an inferometer by measuring the Newtonian gravitational potential the particle produces, the interference pattern would have to be finer than the Planck length and thus indiscernible. Unlike for the electromagnetic field, Bohr's argument does not imply that the gravitational field must be quantized. PMID- 19218439 TI - Odorant receptors at the growth cone are coupled to localized cAMP and Ca2+ increases. AB - A distinctive feature in the topographic organization of the olfactory system in mammals is the dual function of the odorant receptor (OR): it detects odors in the nasal epithelium and plays an instructive role in the axonal convergence of olfactory sensory neurons (OSN) into the olfactory bulb (OB). The latter function is supported by genetic experiments and by the expression of the OR not only on the cilia, but also on the axon termini of the OSN. The signaling pathway coupled to the OR on the cilia is well known and is recognized to involve cAMP and Ca(2+), whereas, until now, nothing was known on the functional characteristics of the OR on the axon termini-growth cone. Here, by analyzing the spatiotemporal dynamics of cAMP and Ca(2+) in living OSN in vitro and in situ, we found that the OR at the growth cone is capable of binding odors and is coupled to cAMP synthesis and Ca(2+) influx through cyclic nucleotide gated (CNG) channels. Furthermore we found that selective odor activation of the OR on the growth cone is followed by nuclear translocation of protein kinase A catalytic subunit. These results define the functional properties of the OR on the growth cone and suggest a potential role of OR activation in axonal convergence and sensory map formation. PMID- 19218441 TI - Pattern formation by dynamically interacting network motifs. AB - Systematic validation of pattern formation mechanisms revealed by molecular studies of development is essentially impossible without mathematical models. Models can provide a compact summary of a large number of experiments that led to mechanism formulation and guide future studies of pattern formation. Here, we realize this program by analyzing a mathematical model of epithelial patterning by the highly conserved EGFR and BMP signaling pathways in Drosophila oogenesis. The model accounts for the dynamic interaction of the feedforward and feedback network motifs that control the expression of Broad, a zinc finger transcription factor expressed in the cells that form the upper part of the respiratory eggshell appendages. Based on the combination of computational analysis and genetic experiments, we show that the model accounts for the key features of wild type pattern formation, correctly predicts patterning defects in multiple mutants, and guides the identification of additional regulatory links in a complex pattern formation mechanism. PMID- 19218442 TI - A Z-DNA sequence reduces slipped-strand structure formation in the myotonic dystrophy type 2 (CCTG) x (CAGG) repeat. AB - All DNA repeats known to undergo expansion leading to human neurodegenerative disease can form one, or several, alternative conformations, including hairpin, slipped strand, triplex, quadruplex, or unwound DNA structures. These alternative structures may interfere with the normal cellular processes of transcription, DNA repair, replication initiation, or polymerase elongation and thereby contribute to the genetic instability of these repeat tracts. We show that (CCTG) x (CAGG) repeats, in the first intron of the ZNF9 gene associated with myotonic dystrophy type 2, form slipped-strand DNA structures in a length-dependent fashion upon reduplexing. The threshold for structure formation on reduplexing is between 36 and 42 repeats in length. Alternative DNA structures also form in (CCTG)(58) x (CAGG)(58) and larger repeat tracts in plasmids at physiological superhelical densities. This represents an example of a sequence that forms slipped-strand DNA from the energy of DNA supercoiling. Moreover, Z-DNA forms in a (TG) x (CA) tract within the complex repeat sequence 5' of the (CCTG)(n) x (CAGG)(n) repeat in the ZNF9 gene. Upon reduplexing, the presence of the flanking sequence containing the Z-DNA-forming tract reduced the extent of slipped-strand DNA formation by 62% for (CCTG)(57) x (CAGG)(57) compared with 58 pure repeats without the flanking sequence. This finding suggests that the Z-DNA-forming sequence in the DM2 gene locus may have a protective effect of reducing the potential for slipped-strand DNA formation in (CCTG)(n) x (CAGG)(n) repeats. PMID- 19218443 TI - Quantitative modeling of the role of surface traps in CdSe/CdS/ZnS nanocrystal photoluminescence decay dynamics. AB - Charge carrier trapping is an important phenomenon in nanocrystal (NC) decay dynamics because it reduces photoluminescence (PL) quantum efficiencies and obscures efforts to understand the interaction of NC excitons with their surroundings. Particularly crucial to our understanding of excitation dynamics in, e.g., multiNC assemblies, would be a way of differentiating between processes involving trap states and those that do not. Direct optical measurement of NC trap state processes is not usually possible because they have negligible transition dipole moments; however, they are known to indirectly affect exciton photoluminescence. Here, we develop a framework, based on Marcus electron transfer theory, to determine NC trap state dynamics from time-resolved NC exciton PL measurements. Our results demonstrate the sensitivity of PL to interfacial dynamics, indicating that the technique can be used as an indirect but effective probe of trap distribution changes. We anticipate that this study represents a step toward understanding how excitons in nanocrystals interact with their surroundings: a quality that must be optimized for their efficient application in photovoltaics, photodetectors, or chemical sensors. PMID- 19218445 TI - Sigma factor mimicry involved in regulation of general stress response. AB - Bacteria have evolved regulatory traits to rapidly adapt to changing conditions. Two principal regulatory mechanisms to modulate gene expression consist of regulation via alternative sigma factors and phosphorylation-dependent response regulators. PhyR represents a recently discovered protein family combining parts of both systems: a sigma factor-like domain of the extracytoplasmic function (ECF) subfamily linked to a receiver domain of a response regulator. Here we investigated the mode of action of this key regulator of general stress response in Methylobacterium extorquens. Our results indicate that PhyR does not act as a genuine sigma factor but instead controls gene expression indirectly through protein-protein interactions. This is evident from the analysis of additional proteins involved in PhyR-dependent gene regulation. We demonstrated that the ECF sigma factor-like domain of PhyR interacts with a protein, designated NepR, upon phosphorylation of the PhyR receiver domain. Using transcriptome analysis and phenotypic assays, we showed that NepR is a negative regulator of PhyR response. Furthermore, we provide biochemical and genetic evidence that NepR exerts this inhibitory effect through sequestration of the ECF sigma factor sigma(EcfG1). Our data support an unprecedented model according to which PhyR acts as a mimicry protein triggering a partner-switching mechanism. Such a regulation of general stress response clearly differs from the two known models operating via sigma(S) and sigma(B). Given the absence of these master regulators and the concomitant conservation of PhyR in Alphaproteobacteria, the novel mechanism presented here is most likely central to the control of general stress response in this large subclass of Proteobacteria. PMID- 19218444 TI - Identification of mRNA splicing factors as the endothelial receptor for carbohydrate-dependent lung colonization of cancer cells. AB - Cell surfaces of epithelial cancer are covered by complex carbohydrates, whose structures function in malignancy and metastasis. However, the mechanism underlying carbohydrate-dependent cancer metastasis has not been defined. Previously, we identified a carbohydrate-mimicry peptide designated I-peptide, which inhibits carbohydrate-dependent lung colonization of sialyl Lewis X expressing B16-FTIII-M cells in E/P-selectin doubly-deficient mice. We hypothesized that lung endothelial cells express an unknown carbohydrate receptor, designated as I-peptide receptor (IPR), responsible for lung colonization of B16-FTIII-M cells. Here, we visualized IPR by in vivo biotinylation, which revealed that the major IPR is a group of 35-kDa proteins. IPR proteins isolated by I-peptide affinity chromatography were identified by proteomics as Ser/Arg-rich alternative pre-mRNA splicing factors or Sfrs1, Sfrs2, Sfrs5, and Sfrs7 gene products. Bacterially expressed Sfrs1 protein bound to B16 FTIII-M cells but not to parental B16 cells. Recombinant Sfrs1 protein bound to a series of fucosylated oligosaccharides in glycan array and plate-binding assays. When anti-Sfrs antibodies were injected intravenously into mice, antibodies labeled a subset of lung capillaries. Anti-Sfrs antibodies inhibited homing of I peptide-displaying phage to the lung colonization of B16-FTIII-M cells in vivo in the mouse. These results strongly suggest that Sfrs proteins are responsible for fucosylated carbohydrate-dependent lung metastasis of epithelial cancers. PMID- 19218446 TI - Scaling and self-organized criticality in proteins I. AB - The complexity of proteins is substantially simplified by regarding them as archetypical examples of self-organized criticality (SOC). To test this idea and elaborate on it, this article applies the Moret-Zebende SOC hydrophobicity scale to the large-scale scaffold repeat protein of the HEAT superfamily, PR65/A. Hydrophobic plasticity is defined and used to identify docking platforms and hinges from repeat sequences alone. The difference between the MZ scale and conventional hydrophobicity scales reflects long-range conformational forces that are central to protein functionality. PMID- 19218447 TI - Synchronization of chaotic early afterdepolarizations in the genesis of cardiac arrhythmias. AB - The synchronization of coupled oscillators plays an important role in many biological systems, including the heart. In heart diseases, cardiac myocytes can exhibit abnormal electrical oscillations, such as early afterdepolarizations (EADs), which are associated with lethal arrhythmias. A key unanswered question is how cellular EADs partially synchronize in tissue, as is required for them to propagate. Here, we present evidence, from computational simulations and experiments in isolated myocytes, that irregular EAD behavior is dynamical chaos. We then show in electrically homogeneous tissue models that chaotic EADs synchronize globally when the tissue is smaller than a critical size. However, when the tissue exceeds the critical size, electrotonic coupling can no longer globally synchronize EADs, resulting in regions of partial synchronization that shift in time and space. These regional partially synchronized EADs then form premature ventricular complexes that propagate into recovered tissue without EADs. This process creates multiple premature ventricular complexes that propagate as [corrected] "shifting" foci resembling polymorphic ventricular tachycardia. Shifting foci encountering shifting repolarization gradients can also develop localized wave breaks leading to reentry and fibrillation. As predicted by the theory, rabbit hearts exposed to oxidative stress (H(2)O(2)) exhibited multiple shifting foci causing polymorphic tachycardia and fibrillation. This mechanism explains how collective cellular behavior integrates at the tissue scale to generate lethal cardiac arrhythmias over a wide range of heart rates. PMID- 19218448 TI - Molecular basis of the interactions between the p73 N terminus and p300: effects on transactivation and modulation by phosphorylation. AB - The transcription factor p73 belongs to the p53 family of proteins and can transactivate a number of target genes in common with p53. Here, we characterized the interaction of the p73 N terminus with four domains of the transcriptional coactivator p300 and with the negative regulator Mdm2 by using biophysical and cellular measurements. We found that, like p53, the N terminus of p73 contained two distinct transactivation subdomains, comprising residues 10-30 and residues 46-67. The p73 N terminus bound weakly to the Taz1, Kix, and IBiD domains of p300 but with submicromolar affinity for Taz2, in contrast to previous reports. We found weaker binding of the p73 N terminus to the p300 domains in vitro correlated with a significant decrease in transactivation activity in a cell line for the QS and T14A mutants, and tighter binding of the phosphomimetic T14D in vitro correlated with an increase in vivo. Further, we found that phosphorylation of T14 increased the affinity of the p73 N terminus for Taz2 10-fold. The phosphomimetic p73alpha T14D caused increased levels of transactivation. PMID- 19218449 TI - Role of COX-2 in epithelial-stromal cell interactions and progression of ductal carcinoma in situ of the breast. AB - Epithelial-stromal cell interactions have an important role in breast tumor progression, but the molecular mechanisms underlying these effects are just beginning to be understood. We previously described that fibroblasts promote, whereas normal myoepithelial cells inhibit, the progression of ductal carcinoma in situ (DCIS) to invasive breast carcinomas by using a xenograft model of human DCIS. Here, we report that the tumor growth and progression-promoting effects of fibroblasts are at least in part due to increased COX-2 expression in tumor epithelial cells provoked by their interaction with fibroblasts. Up-regulation of COX-2 in DCIS xenografts resulted in increased VEGF and MMP14 expression, which may contribute to the larger weight and invasive histology of COX-2-expressing tumors. Administration of celecoxib, a selective COX-2 inhibitor, to tumor bearing mice decreased xenograft tumor weight and inhibited progression to invasion. Coculture of fibroblasts with DCIS epithelial cells enhanced their motility and invasion, and this change was associated with increased MMP14 expression and MMP9 protease activity. We identified the NF-kappaB pathway as one of the mediators of stromal fibroblast-derived signals regulating COX-2 expression in tumor epithelial cells. Inhibition of NF-kappaB and COX-2 activity and down-regulation of MMP9 expression attenuated the invasion-promoting effects of fibroblasts. These findings support a role for COX-2 in promoting the progression of DCIS to invasive breast carcinomas, and suggest that therapeutic targeting of the NF-kappaB and prostaglandin signaling pathways might be used for the treatment and prevention of breast cancer. PMID- 19218450 TI - Enhanced selection of FoxP3+ T-regulatory cells protects CTLA-4-deficient mice from CNS autoimmune disease. AB - It is generally acknowledged that cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4/CD152) plays a pivotal role in the regulation of T-cell activation and the establishment of self-tolerance in the periphery. CTLA-4-deficient (CTLA-4KO) mice develop a lymphoproliferative disorder and die within 4 weeks of birth, suggesting a role for CTLA-4 in T-cell homeostasis or the development and activity of T-regulatory (Treg) cells. To study the role of CTLA-4 in the control of experimental autoimmune encephalomyelitis (EAE), we have generated a CTLA-4KO mouse in which >90% of all CD4(+) T cells bear a Vbeta8.2 transgenic T-cell receptor that is specific for myelin basic protein peptide Ac1-9 (ASQKRPSQR). These mice do not develop spontaneous lymphoproliferative disease or EAE and are resistant to disease induction. This correlates with a higher frequency of functional FoxP3(+) Treg cells in the spleen and thymus of CTLA-4KO mice. The absence of CTLA-4-mediated suppression of CD28 signaling resulted in the early expression of FoxP3 on double-positive cells in the thymic cortex. We conclude that CTLA-4 is not essential for the peripheral function of FoxP3(+) Treg cells but plays a pivotal role in their thymic selection. PMID- 19218451 TI - LapD is a bis-(3',5')-cyclic dimeric GMP-binding protein that regulates surface attachment by Pseudomonas fluorescens Pf0-1. AB - The second messenger cyclic dimeric GMP (c-di-GMP) regulates surface attachment and biofilm formation by many bacteria. For Pseudomonas fluorescens Pf0-1, c-di GMP impacts the secretion and localization of the adhesin LapA, which is absolutely required for stable surface attachment and biofilm formation by this bacterium. In this study we characterize LapD, a unique c-di-GMP effector protein that controls biofilm formation by communicating intracellular c-di-GMP levels to the membrane-localized attachment machinery via its periplasmic domain. LapD contains degenerate and enzymatically inactive diguanylate cyclase and c-di-GMP phosphodiesterase (EAL) domains and binds to c-di-GMP through a degenerate EAL domain. We present evidence that LapD utilizes an inside-out signaling mechanism: binding c-di-GMP in the cytoplasm and communicating this signal to the periplasm via its periplasmic domain. Furthermore, we show that LapD serves as the c-di-GMP receptor connecting environmental modulation of intracellular c-di-GMP levels by inorganic phosphate to regulation of LapA localization and thus surface commitment by P. fluorescens. PMID- 19218452 TI - Interaction forces and adhesion of supported myelin lipid bilayers modulated by myelin basic protein. AB - Force-distance measurements between supported lipid bilayers mimicking the cytoplasmic surface of myelin at various surface coverages of myelin basic protein (MBP) indicate that maximum adhesion and minimum cytoplasmic spacing occur when each negative lipid in the membrane can bind to a positive arginine or lysine group on MBP. At the optimal lipid/protein ratio, additional attractive forces are provided by hydrophobic, van der Waals, and weak dipolar interactions between zwitterionic groups on the lipids and MBP. When MBP is depleted, the adhesion decreases and the cytoplasmic space swells; when MBP is in excess, the bilayers swell even more. Excess MBP forms a weak gel between the surfaces, which collapses on compression. The organization and proper functioning of myelin can be understood in terms of physical noncovalent forces that are optimized at a particular combination of both the amounts of and ratio between the charged lipids and MBP. Thus loss of adhesion, possibly contributing to demyelination, can be brought about by either an excess or deficit of MBP or anionic lipids. PMID- 19218454 TI - Exploring the likelihood and mechanism of a climate-change-induced dieback of the Amazon rainforest. AB - We examine the evidence for the possibility that 21st-century climate change may cause a large-scale "dieback" or degradation of Amazonian rainforest. We employ a new framework for evaluating the rainfall regime of tropical forests and from this deduce precipitation-based boundaries for current forest viability. We then examine climate simulations by 19 global climate models (GCMs) in this context and find that most tend to underestimate current rainfall. GCMs also vary greatly in their projections of future climate change in Amazonia. We attempt to take into account the differences between GCM-simulated and observed rainfall regimes in the 20th century. Our analysis suggests that dry-season water stress is likely to increase in E. Amazonia over the 21st century, but the region tends toward a climate more appropriate to seasonal forest than to savanna. These seasonal forests may be resilient to seasonal drought but are likely to face intensified water stress caused by higher temperatures and to be vulnerable to fires, which are at present naturally rare in much of Amazonia. The spread of fire ignition associated with advancing deforestation, logging, and fragmentation may act as nucleation points that trigger the transition of these seasonal forests into fire dominated, low biomass forests. Conversely, deliberate limitation of deforestation and fire may be an effective intervention to maintain Amazonian forest resilience in the face of imposed 21st-century climate change. Such intervention may be enough to navigate E. Amazonia away from a possible "tipping point," beyond which extensive rainforest would become unsustainable. PMID- 19218453 TI - Early and sustained innate immune response defines pathology and death in nonhuman primates infected by highly pathogenic influenza virus. AB - The mechanisms responsible for the virulence of the highly pathogenic avian influenza (HPAI) and of the 1918 pandemic influenza virus in humans remain poorly understood. To identify crucial components of the early host response during these infections by using both conventional and functional genomics tools, we studied 34 cynomolgus macaques (Macaca fascicularis) to compare a 2004 human H5N1 Vietnam isolate with 2 reassortant viruses possessing the 1918 hemagglutinin (HA) and neuraminidase (NA) surface proteins, known conveyors of virulence. One of the reassortants also contained the 1918 nonstructural (NS1) protein, an inhibitor of the host interferon response. Among these viruses, HPAI H5N1 was the most virulent. Within 24 h, the H5N1 virus produced severe bronchiolar and alveolar lesions. Notably, the H5N1 virus targeted type II pneumocytes throughout the 7 day infection, and induced the most dramatic and sustained expression of type I interferons and inflammatory and innate immune genes, as measured by genomic and protein assays. The H5N1 infection also resulted in prolonged margination of circulating T lymphocytes and notable apoptosis of activated dendritic cells in the lungs and draining lymph nodes early during infection. While both 1918 reassortant viruses also were highly pathogenic, the H5N1 virus was exceptional for the extent of tissue damage, cytokinemia, and interference with immune regulatory mechanisms, which may help explain the extreme virulence of HPAI viruses in humans. PMID- 19218455 TI - Oceanic acidification affects marine carbon pump and triggers extended marine oxygen holes. AB - Rising atmospheric CO(2) levels will not only drive future global mean temperatures toward values unprecedented during the whole Quaternary but will also lead to massive acidification of sea water. This constitutes by itself an anthropogenic planetary-scale perturbation that could significantly modify oceanic biogeochemical fluxes and severely damage marine biota. As a step toward the quantification of such potential impacts, we present here a simulation-model based assessment of the respective consequences of a business-as-usual fossil fuel-burning scenario where a total of 4,075 Petagrams of carbon is released into the atmosphere during the current millennium. In our scenario, the atmospheric pCO(2) level peaks at approximately 1,750 microatm in the year 2200 while the sea surface pH value drops by >0.7 units on global average, inhibiting the growth of marine calcifying organisms. The study focuses on quantifying 3 major concomitant effects. The first one is a significant (climate-stabilizing) negative feedback on rising pCO(2) levels as caused by the attenuation of biogenic calcification. The second one is related to the biological carbon pump. Because mineral ballast, notably CaCO(3), is found to play a dominant role in carrying organic matter through the water column, a reduction of its export fluxes weakens the strength of the biological carbon pump. There is, however, a third effect with severe consequences: Because organic matter is oxidized in shallow waters when mineral ballast fluxes weaken, oxygen holes (hypoxic zones) start to expand considerably in the oceans in our model world--with potentially harmful impacts on a variety of marine ecosystems. PMID- 19218456 TI - Massively parallel sequencing identifies the gene Megf8 with ENU-induced mutation causing heterotaxy. AB - Forward genetic screens with ENU (N-ethyl-N-nitrosourea) mutagenesis can facilitate gene discovery, but mutation identification is often difficult. We present the first study in which an ENU-induced mutation was identified by massively parallel DNA sequencing. This mutation causes heterotaxy and complex congenital heart defects and was mapped to a 2.2-Mb interval on mouse chromosome 7. Massively parallel sequencing of the entire 2.2-Mb interval identified 2 single-base substitutions, one in an intergenic region and a second causing replacement of a highly conserved cysteine with arginine (C193R) in the gene Megf8. Megf8 is evolutionarily conserved from human to fruit fly, and is observed to be ubiquitously expressed. Morpholino knockdown of Megf8 in zebrafish embryos resulted in a high incidence of heterotaxy, indicating a conserved role in laterality specification. Megf8(C193R) mouse mutants show normal breaking of symmetry at the node, but Nodal signaling failed to be propagated to the left lateral plate mesoderm. Videomicroscopy showed nodal cilia motility, which is required for left-right patterning, is unaffected. Although this protein is predicted to have receptor function based on its amino acid sequence, surprisingly confocal imaging showed it is translocated into the nucleus, where it is colocalized with Gfi1b and Baf60C, two proteins involved in chromatin remodeling. Overall, through the recovery of an ENU-induced mutation, we uncovered Megf8 as an essential regulator of left-right patterning. PMID- 19218457 TI - Evidence that family 35 carbohydrate binding modules display conserved specificity but divergent function. AB - Enzymes that hydrolyze complex carbohydrates play important roles in numerous biological processes that result in the maintenance of marine and terrestrial life. These enzymes often contain noncatalytic carbohydrate binding modules (CBMs) that have important substrate-targeting functions. In general, there is a tight correlation between the ligands recognized by bacterial CBMs and the substrate specificity of the appended catalytic modules. Through high-resolution structural studies, we demonstrate that the architecture of the ligand binding sites of 4 distinct family 35 CBMs (CBM35s), appended to 3 plant cell wall hydrolases and the exo-beta-D-glucosaminidase CsxA, which contributes to the detoxification and metabolism of an antibacterial fungal polysaccharide, is highly conserved and imparts specificity for glucuronic acid and/or Delta4,5 anhydrogalaturonic acid (Delta4,5-GalA). Delta4,5-GalA is released from pectin by the action of pectate lyases and as such acts as a signature molecule for plant cell wall degradation. Thus, the CBM35s appended to the 3 plant cell wall hydrolases, rather than targeting the substrates of the cognate catalytic modules, direct their appended enzymes to regions of the plant that are being actively degraded. Significantly, the CBM35 component of CsxA anchors the enzyme to the bacterial cell wall via its capacity to bind uronic acid sugars. This latter observation reveals an unusual mechanism for bacterial cell wall enzyme attachment. This report shows that the biological role of CBM35s is not dictated solely by their carbohydrate specificities but also by the context of their target ligands. PMID- 19218458 TI - A mitochondrial DNA mutation linked to colon cancer results in proton leaks in cytochrome c oxidase. AB - An increasing number of cancer types have been found to be linked to specific mutations in the mitochondrial DNA, which result in specific structural changes of the respiratory enzyme complexes. In this study, we have investigated the effect of 2 such mutations identified in colon cancer patients, leading to the amino acid substitutions Ser458Pro and Gly125Asp in subunit I of cytochrome c oxidase (complex IV) [Greaves et al. (2006) Proc Natl Acad Sci USA 103:714-719]. We introduced these mutations in Rhodobacter sphaeroides, which carries an oxidase that serves as a model of the mitochondrial counterpart. The lack of expression of the former variant indicates that the amino acid substitution results in severely altered overall structure of the enzyme. The latter mutation (Gly171Asp in the bacterial oxidase) resulted in a structurally intact enzyme, but with reduced activity (approximately 30%), mainly due to slowed reduction of the redox site heme a. Furthermore, even though the Gly171Asp CytcO pumps protons, an intrinsic proton leak was identified, which would lead to a decreased overall energy-conversion efficiency of the respiratory chain, and would also perturb transport processes such as protein, ion, and metabolite trafficking. Furthermore, the specific leak may act to alter the balance between the electrical and chemical components of the proton electrochemical gradient. PMID- 19218459 TI - Control of Arabidopsis meristem development by thioredoxin-dependent regulation of intercellular transport. AB - Cell-to-cell transport in plants occurs through cytoplasmic channels called "plasmodesmata" and is regulated by developmental and environmental factors. Callose deposition modulates plasmodesmal transport in vivo, but little is known about the mechanisms that regulate this process. Here we report a genetic approach to identify mutants affecting plasmodesmal transport. We isolated 5 mutants, named gfp arrested trafficking (gat), affected in GFP unloading from the phloem into the meristem. gat1 mutants were seedling lethal and carried lesions in an m-type thioredoxin that is expressed in non-green plastids of meristems and organ primordia. Callose and hydrogen peroxide accumulated in gat1 mutants, and WT plants subjected to oxidative conditions phenocopied the gat1 trafficking defects. Ectopic expression of GAT1 in mature leaves increased plasmodesmal permeability and led to a delay in senescence and flowering time. We propose a role for the GAT1 thioredoxin in the redox regulation of callose deposition and symplastic permeability that is essential for meristem maintenance in Arabidopsis. PMID- 19218460 TI - A manganese transporter, BB0219 (BmtA), is required for virulence by the Lyme disease spirochete, Borrelia burgdorferi. AB - Borrelia burgdorferi (Bb), the causative agent of Lyme disease, is transmitted to mammalian hosts through an arthropod (tick) vector. To establish infection, Bb must acquire essential nutrients, including transition metals, from its mammalian and tick hosts. Thus far, no metal transporter has been identified in Bb. Here, we report the identification of the first metal transporter, BmtA (BB0219), in Bb. BmtA-deficient mutants of virulent Bb were readily generated, and the mutants grew slightly slower than wild-type Bb in vitro. However, BmtA mutants were sensitive to the chelating actions of EDTA, suggesting a role for BmtA in metal utilization. Intracellular accumulation of manganese (Mn) was substantially diminished in the bmtA mutant, indicating that BmtA was operative in Mn uptake. Given that BmtA lacks homology to any known Mn transporter, we postulate that BmtA is part of a novel mechanism for Mn acquisition by a bacterial pathogen. BmtA also was essential to the infectious life cycle of Bb in ticks and mammals, thereby qualifying BmtA as a new borrelial virulence factor. In addition, the bmtA mutant was sensitive to treatment with t-butyl hydroperoxide, implying that BmtA, and thus Mn, is important to Bb for detoxifying reactive oxygen species, including those potentially liberated by immune effector cells during the innate immune response. Our discovery of the first molecule involved in metal transport in Bb provides a foundation for further elucidating metal homeostasis in this important human pathogen, which may lead to new strategies for thwarting Lyme disease. PMID- 19218461 TI - Connexin 32 increases the proliferative response of Schwann cells to neuregulin-1 (Nrg1). AB - Connexin 32 (Cx32), a gap junction protein, is found within the para-nodal region and Schmidt-Lanterman incisures of myelinating Schwann cells (SCs). In developing and regenerating peripheral nerves, pro-myelinating SCs express Cx32 mRNA and protein in conjunction with the expression of myelin specific genes. Neuregulin-1 (Nrg1), a member of the neuregulin family of growth factors, controls SC proliferation and differentiation depending on the cellular environment and the particular stage of SC maturation. Primary cultures of purified SCs from newborn mouse sciatic nerve were used to characterize both the role of Nrg1 in the expression of Cx32 and, conversely, the role of Cx32 in SC responsiveness to Nrg1. Glial growth factor 2, an isoform of Nrg1, up-regulated Cx32 in both proliferating and non-proliferating SCs. However, SCs from Cx32-KO mice exhibited a significantly smaller mitogenic response to glial growth factor 2. Electrical coupling between Cx32-KO SCs did not differ from that between WT SCs, indicating the presence of other connexins. These results suggest a link between Cx32 expression and Nrg1 regulation of SC proliferation that does not involve Cx32 mediated intercellular communication. PMID- 19218462 TI - Phosphorylation of Rap1GAP, a striatally enriched protein, by protein kinase A controls Rap1 activity and dendritic spine morphology. AB - Protein kinase A (PKA)-dependent signaling cascades play an important role in mediating the effects of dopamine and other neurotransmitters in striatal medium spiny neurons. We have identified a prominent striatal PKA substrate as Rap1 GTPase activating protein (Rap1GAP), a negative regulator of Rap1 signaling. Although present throughout the brain, Rap1GAP is enriched in striatal medium spiny neurons and is phosphorylated by PKA at Ser-441 and Ser-499 in response to activation of D1 dopamine receptors. Phosphorylation of Rap1GAP is associated with inhibition of GAP activity, as demonstrated by increased Rap1 activity in striatal neurons. Phosphorylation of Rap1GAP is also associated with decreased [corrected] dendritic spine head size in cultured neurons. These findings suggest that phosphorylation of Rap1GAP by PKA plays an important role in striatal neurons by modulating Rap1 actions. PMID- 19218464 TI - Undergraduate nursing students writing therapeutic letters to families: an educational strategy. AB - Writing therapeutic letters to families is discussed in this article as an educational strategy encouraging students to think reflectively about family nursing. At the University of Kalmar, Sweden, undergraduate nursing students in a primary care module interviewed families using the Calgary Family Assessment Model and wrote therapeutic letters to these families. This article describes (a) the examination process, which was the context for writing therapeutic letters, (b) results of analyses of the letters, and (c) student's post-examination evaluation comments. Results indicate that most students needed encouragement to focus on the family's strengths and resources instead of focusing on own feelings or problems they perceived the family as having. Students also needed support in relinquishing their hierarchical role of "expert nurse." Students' evaluation comments showed that writing therapeutic letters provided students with opportunities to reflect about the connections between family nursing theory and the family itself. PMID- 19218465 TI - The past and future of therapeutic letters: family suffering and healing words. AB - This article explores the technological implications of the translation of therapeutic letters to therapeutic e-mail communication. The history of letters is juxtaposed with the currency with which the written language locates itself in today's methods and patterns of communication. Implications of such a translation, inherent with gifts and limitations, all deeply embedded in a historical context, are explored. The future of therapeutic letters continues to be located in their utility and ultimately their capacity to offer healing and helpful words to families in suffering. PMID- 19218463 TI - Receptor-mediated phagocytosis elicits cross-presentation in nonprofessional antigen-presenting cells. AB - In cross-presentation by dendritic cells (DCs), internalized proteins are retrotranslocated into the cytosol, degraded by the proteasome, and the generated antigenic peptides bind to MHC class I molecules for presentation on the cell surface. Endoplasmic reticulum (ER) contribution to phagosomal membranes is thought to provide antigen access to the ER-associated degradation (ERAD) machinery, allowing cytosolic dislocation. Because the ERAD pathway is present in all cell types and exogenous antigens encounter an ER-containing compartment during phagocytosis, we postulated that forcing phagocytosis in cell types other than DCs would render them competent for cross-presentation. Indeed, FcRgammaIIA expression endowed 293T cells with the capacity for both phagocytosis and ERAD mediated cross-presentation of an antigen provided as an immune complex. The acquisition of this ability by nonprofessional antigen-presenting cells suggests that a function potentially available in all cell types has been adapted by DCs for presentation of exogenous antigens by MHC class I molecules. PMID- 19218466 TI - Therapeutic letters and the family nursing unit: a legacy of advanced nursing practice. AB - This article focuses on the history of the use of therapeutic letters in the clinical scholarship of the Family Nursing Unit at the University of Calgary and offers examples of a variety of therapeutic letters written to families experiencing illness suffering. A case study from the research of Moules (2000, 2002) is offered to further illustrate the usefulness of therapeutic letters as a family nursing intervention. PMID- 19218467 TI - Multivariable models in biobehavioral research. AB - OBJECTIVE: To review contemporary multivariable modeling and statistical reporting practices in psychosomatic and behavioral medicine research. METHODS: A random sample of 40 original research articles involving multivariable models was obtained from the 2005 volumes of four of the leading psychosomatic and behavioral medicine research journals. A random comparison sample was obtained from the 2005 volumes of four of the leading general medical and psychiatric journals. Multivariable modeling and reporting practices were systematically coded. The evaluation focused primarily on issues raised in 2004 Statistical Corner article by Babyak. RESULTS: Deficiencies were found in a large proportion of the articles published in psychosomatic and behavioral medicine journals. The single most common problem was a lack of clear information, or any information at all, about important aspects of the statistical methods. Other frequent problems included post hoc selection of variables, lack of clear rationales and well specified roles for selected variables, inadequate information about models as a whole (e.g., goodness of fit), failure to test model assumptions, and lack of model validation. Overfitting of multivariable models was the exception rather than the rule, but still a significant problem. CONCLUSIONS: There is room for improvement in the use and reporting of multivariable models in psychosomatic and behavioral medicine research journals. These problems can be overcome by adopting best statistical practices, such as those recommended by Psychosomatic Medicine's statistical guidelines and by authoritative guidebooks on statistical reporting practices. PMID- 19218470 TI - The hazards of transplant tourism. PMID- 19218471 TI - Pulse pressure and presence of coronary artery calcification. AB - BACKGROUND: Coronary calcification (CAC) is found in early stages of CKD. Pulse pressure (PP) predicts CAC in dialysis patients. This study evaluates the accuracy of PP in predicting CAC in patients not yet on dialysis (CKD patients). METHODS: CKD patients (n = 388) underwent coronary calcium score (CAC score) and abdominal x-ray (n = 128) for estimating aorta calcification (AAC). Biochemistry and PP were measured every 3 and 6 months in patients with stage 4 to 5 and 2 to 3 CKD, respectively. The accuracy of PP and AAC was assessed by receiver operating characteristics analysis. RESULTS: PP correlated with CAC score in the whole cohort and in patients with stages 2 to 3 and stages 4 to 5 CKD. PP >60 mmHg predicted CAC score >0 (OR: 2.14; P < 0.001), > or =100 (OR: 2.92; P < 0.001), > or =400 (OR: 6.17; P < 0.001) after multivariable adjustment. Area under the curve (AUC) was 0.626 for CAC score >0, 0.676 for score >100, and 0.746 for score >400. PP >60 mmHg reduced the rate of event-free survival. AAC was found in 58% of patients and correlated with CAC score. AUC was 0.628 for CAC score >0, 0.652 for score >100, 0.831 for score >400. CONCLUSION: PP may identify CKD patients with subclinical CAC who need further evaluation. Accuracy of PP and AAC is nearly similar in predicting CAC. High PP indicates vessel wall alterations leading to adverse outcome. PMID- 19218472 TI - Perception of indications for nephrology referral among internal medicine residents: a national online survey. AB - BACKGROUND AND OBJECTIVES: Many patients with chronic kidney disease (CKD) are seen by primary care physicians who may not be aware of indications or benefits of timely nephrologist referral. Late referral to a nephrologist may lead to suboptimal pre-end stage renal disease care and greater mortality. It is not known whether current postgraduate training adequately prepares a future internist in this aspect of CKD management. DESIGN, SETTING, PARTICIPANTS, AND MEASUREMENTS: The authors performed an online questionnaire survey of internal medicine residents in the United States to determine their perceptions of indications for nephrology referral in CKD management. RESULTS: Four hundred seventy-nine residents completed the survey with postgraduate year (PGY) distribution of 166 PGY 1,187 PGY 2 and 126 PGY 3. Few residents chose nephrology referral for proteinuria (45%), uncontrolled hypertension (64%), or hyperkalemia (26%). Twenty-eight percent of the residents considered consulting a nephrologist for anemia of CKD, whereas 45% would do so for bone disorder of CKD. Most of the residents would involve a nephrologist at glomerular filtration rate (GFR) <30 ml/min/1.73 m(2) (90%) and for rapid decline in GFR (79%). Many residents would refer a patient for dialysis setup at GFR 15 to 30 ml/min/1.73 m(2) (59%); however, 18% would do so at GFR <15 ml/min/1.73 m(2). Presence of CKD clinic experience or an in-house nephrology fellowship program did not considerably change these perceptions. CONCLUSIONS: Results show that internal medicine residents have widely differing perceptions of indications for nephrology referral. Educational efforts during residency training to raise awareness and benefits of early referral may improve CKD management by facilitating better collaboration between internist and nephrologist. PMID- 19218473 TI - Optimal method of coronary revascularization in patients receiving dialysis: systematic review. AB - BACKGROUND AND OBJECTIVES: Patients receiving dialysis have a high burden of cardiovascular disease. Some receive coronary artery revascularization but the optimal method is controversial. DESIGN, SETTING, PARTICIPANTS, & MEASUREMENTS: The authors reviewed any randomized controlled trial or cohort study of 10 or more patients receiving maintenance dialysis which compared coronary artery bypass graft (CABG) to percutaneous intervention (PCI) for revascularization of the coronary arteries. The primary outcomes were short-term (30 d or in-hospital) and long-term (at least 1 year) mortality. RESULTS: Seventeen studies were found. There were no randomized trials: all were retrospective cohort studies from years 1977 to 2002. There were some baseline differences between the groups receiving CABG compared with those receiving PCI, and most studies did not consider results adjusted for such characteristics. Given the variability among studies and their methodological limitations, few definitive conclusions about the optimal method of revascularization could be drawn. In an exploratory meta-analysis, short-term mortality was higher after CABG compared to PCI. A substantial number of patients died over a subsequent 1 to 5 yr, with no difference in mortality after CABG compared to PCI. CONCLUSIONS: Although decisions about the optimal method of coronary artery revascularization in dialysis patients are undertaken routinely, it was surprising to see how few data has been published in this regard. Additional research will help inform physician and patient decisions about coronary artery revascularization. PMID- 19218474 TI - Arterial hypertension induced by erythropoietin and erythropoiesis-stimulating agents (ESA). AB - This review summarizes the evidence for a hypertensinogenic effect of Erythropoietin (Epo) in normal human subjects and predialysis, hemodialysis, and continuous ambulatory peritoneal dialysis (CAPD) patients. The possible mechanisms of Epo-induced hypertension are examined with in vivo animal and in vitro data, as well as pathophysiological human studies in both normal subjects and CKD patients. The evidence for a hypertensinogenic effect of erythropoiesis stimulating agents (ESAs) in normal subjects, predialysis CKD, hemodialysis, and CAPD patients is compelling. Epo increases BP directly and notably independently of its erythropoietic effect and its effect on blood rheology. The potential for the development of future agents that might act as specific stimulators of erythropoiesis, devoid of direct hemodynamic side effects is underscored. PMID- 19218475 TI - Calcineurin inhibitor nephrotoxicity. AB - The use of the calcineurin inhibitors cyclosporine and tacrolimus led to major advances in the field of transplantation, with excellent short-term outcome. However, the chronic nephrotoxicity of these drugs is the Achilles' heel of current immunosuppressive regimens. In this review, the authors summarize the clinical features and histologic appearance of both acute and chronic calcineurin inhibitor nephrotoxicity in renal and nonrenal transplantation, together with the pitfalls in its diagnosis. The authors also review the available literature on the physiologic and molecular mechanisms underlying acute and chronic calcineurin inhibitor nephrotoxicity, and demonstrate that its development is related to both reversible alterations and irreversible damage to all compartments of the kidneys, including glomeruli, arterioles, and tubulo-interstitium. The main question--whether nephrotoxicity is secondary to the actions of cyclosporine and tacrolimus on the calcineurin-NFAT pathway--remains largely unanswered. The authors critically review the current evidence relating systemic blood levels of cyclosporine and tacrolimus to calcineurin inhibitor nephrotoxicity, and summarize the data suggesting that local exposure to cyclosporine or tacrolimus could be more important than systemic exposure. Finally, other local susceptibility factors for calcineurin inhibitor nephrotoxicity are reviewed, including variability in P-glycoprotein and CYP3A4/5 expression or activity, older kidney age, salt depletion, the use of nonsteroidal anti-inflammatory drugs, and genetic polymorphisms in genes like TGF-beta and ACE. Better insight into the mechanisms underlying calcineurin inhibitor nephrotoxicity might pave the way toward more targeted therapy or prevention of calcineurin inhibitor nephrotoxicity. PMID- 19218476 TI - Sexual abuse: a journal of research and treatment performance indicators for 2007. PMID- 19218477 TI - Psychometric properties of the Marlowe-Crowne social desirability scale with adult male sexual offenders. AB - This research project investigates the factor structure, reliability, and validity of the Marlowe-Crowne Social Desirability Scale (MCSDS) when used with a sample of convicted, adult male sexual offenders. Results confirm the two-factor structure (Denial and Attribution) originally proposed by the authors (Study 1, n = 247). Results also reveal that the full-scale MCSDS has strong internal consistency estimates (Study 1), discriminant and convergent validity (Study 2, n = 91), and test-retest reliability over a 3-week period (Study 3, n = 74). Information is provided to aid evaluators' interpretation of MCSDS full-scale and factor scores when used with male adult sexual offenders. PMID- 19218478 TI - Assessing the implicit beliefs of sexual offenders using the implicit relational assessment procedure: a first study. AB - Researchers have proposed that the cognitive distortions of sexual offenders are underpinned by a number of implicit cognitive processes termed implicit theories. Until recently, however, the implicit theory hypothesis has received little empirical support due to broader limitations with standard forensic assessment procedures. The current research aimed to determine whether a new assessment methodology, the Implicit Relational Assessment Procedure (IRAP), could provide further evidence for Ward and Keenan's (1999) children as sexual beings implicit theory. The results indicated that the IRAP was significantly more effective at identifying core implicit differences between sexual offenders against children and nonoffenders than a cognitive distortion questionnaire. Furthermore, although both groups demonstrated an overall response bias towards adults as sexual and children as nonsexual, this bias was significantly impaired in the sexual offender group. The findings are discussed in relation to previous implicit theory research and recommendations for the development of implicit methodologies are made. PMID- 19218479 TI - Psychological profiles of internet sexual offenders: comparisons with contact sexual offenders. AB - A sample of 505 Internet sex offenders and 526 contact sex offenders were compared on a range of psychological measures relating to offense-supportive beliefs, empathic concern, interpersonal functioning, and emotional management. Internet offenders could be successfully discriminated from contact offenders on 7 out of 15 measures. Contact offenders were found to have significantly more victim empathy distortions and cognitive distortions than Internet offenders. Internet offenders were found to have significantly higher identification with fictional characters than contact offenders. Further analysis indicated that an increase in scores on scales of fantasy, underassertiveness, and motor impulsivity were predictive of an Internet offense type. An increase in scores of scales of locus of control, perspective taking, empathic concern, overassertiveness, victim empathy distortions, cognitive distortions, and cognitive impulsivity were found to be predictive of a contact offense type. These findings are discussed in the context of the etiology of sexual offending. PMID- 19218480 TI - Facial and prosodic affect recognition among pedophilic and nonpedophilic criminal child molesters. AB - The present study compared facial and prosodic affect recognition abilities among pedophilic and nonpedophilic child molesters and community-dwelling controls. Pedophilic child molesters are characterized by primary sexual interest in prepubescent children, whereas nonpedophilic child molesters are characterized by offending against children despite being primarily sexually attracted to adults. The results showed that nonpedophilic child molesters made more errors in recognizing both facial and prosodic affect, performing more poorly than both controls and pedophilic child molesters. These findings are consistent with greater psychopathic tendencies among nonpedophilic molesters as well as with prior findings of smaller amygdala volume among child molesters. PMID- 19218481 TI - Individual differences in the propensity for partner sexual coercion. AB - Lalumiere, Harris, Quinsey, and Rice (2005) proposed a three-path model (psychopathy, young male syndrome, and competitive disadvantage) of the development of sexually coercive behavior, but none of these individual difference characteristics have been tested among partner rapists. Using a community sample in Study 1, the authors find that psychopathy is the only significant predictor of self-reported propensity for partner sexual coercion. This model is tested in Study 2 by comparing convicted partner rapists, nonsexual partner assaulters, and heterosexual child molesters. One third of partner rapists are psychopaths, and their psychopathy scores are no different from those found in correctional samples. Partner rapists have an average IQ, providing further evidence that competitive disadvantage is less characteristic of partner rapists. There is some indication that partner rapists desist with age. The authors discuss these findings in light of recent findings that implicate cuckoldry risk in partner sexual assault. PMID- 19218482 TI - Transgenic zebrafish reporter lines reveal conserved Toll-like receptor signaling potential in embryonic myeloid leukocytes and adult immune cell lineages. AB - The immune response of a host to an invading pathogen is dependent on the capacity of its immune cell compartment to recognize highly conserved pathogen components using an ancient class of pattern recognition receptors known as Toll like receptors (TLRs). Initiation of TLR-mediated signaling results in the induction of proinflammatory cytokines that help govern the scale and duration of any ensuing response. Specificity for TLR signaling is, in part, a result of the differential recruitment of intracellular adaptor molecules. Of these, MyD88 is required for the majority of TLR signaling. Zebrafish have been shown to possess TLRs and adaptor molecules throughout early development, including MyD88, strongly suggesting conservation of this ancient defense mechanism. However, information about which embryonic cells/tissues possess this conserved signaling potential is lacking. To help define which embryonic cells, in particular, those of the innate immune system, have the potential for MyD88-dependent, TLR-mediated signaling, we generated transgenic reporter lines using regulatory elements of the myd88 gene to drive the fluorescent reporters enhanced GFP and Discosoma red fluorescent protein 2 within live zebrafish. These lines possess fluorescently marked cells/tissues consistent with endogenous myd88 expression, including a subset of myeloid leukocytes. These innate immune cells were confirmed to express other TLR adaptors including Mal, trif, and Sarm. Live wound-healing and infection assays validated the potential of these myd88-expressing leukocytes to participate in immune responses. These lines will provide a valuable resource for further resolving the contribution of MyD88 to early vertebrate immunity. PMID- 19218484 TI - Magnitude of type 1 diabetes self-management in youth: health care needs diabetes educators. AB - The purpose of this article is to demonstrate the complexity of the type 1 diabetes regimen and to highlight the essential role of the diabetes educator in safely training and implementing the myriad skills in a developmentally appropriate manner for children and adolescents. A review of literature and a task analysis were preformed and suggest that the complexity of the regimen is often not adequately addressed. Reviewed research assessed the regimen using measures with on average about 25 items while the task analysis contains over 600 tasks. The article discusses implications for clinical practice, including implications for measurement in research, targeting of interventions by diabetes educators, and the gradual transfer of regimen control to youth. It is argued that given the magnitude of the self-management task, education cannot be accomplished in the limited time that general practice physicians, pediatricians, or endocrinology specialists can spend with each patient. It is concluded that youth must be helped to internalize the importance of the regimen tasks and that transfer of these tasks to youth requires a developmentally sensitive approach to education. Diabetes educators serve an essential role in which they help young patients and their parents manage and master this overwhelming experience through promoting youth's involvement in tasks when full responsibility is not yet appropriate. The regimen is too complex for youth to undertake self-management without multidisciplinary support. PMID- 19218483 TI - IFN-gamma-indoleamine-2,3 dioxygenase acts as a major suppressive factor in 4-1BB mediated immune suppression in vivo. AB - It has been reported that 4-1BB triggering in vivo selectively suppressed the recall response of staphylococcal enterotoxin A (SEA)-specific CD4(+) T cells, in which CD8(+) T-derived TGF-beta was involved. Here, we have examined an alternative mechanism for the 4-1BB-mediated CD4(+) T suppression, as the neutralization of TGF-beta is only effective in rescuing the SEA-specific recall response at high cellular concentrations. We show that this selective suppression of CD4(+) T cells by 4-1BB triggering in vivo is mediated mainly by induction of indoleamine 2,3-dioxygenase (IDO) in an IFN-gamma-dependent manner. SEA-specific CD4(+) T responses were suppressed partly by TGF-beta-expressing CD8(+) T cells, particularly CD11c(+)CD8(+) T cells, but strongly inhibited by dendritic cells (DCs) expressing IDO. IFN-gamma that increased IDO in DCs was produced primarily from CD11c(+)CD8(+) T cells, which were expanded selectively by 4-1BB stimulation. CD4(+), CD8(+), and plasmacytoid DCs exerted a similar suppressive activity toward the SEA-specific CD4(+) T cells. Neutralization of IFN-gamma or IDO activity in vivo largely reversed the 4-1BB-mediated CD4(+) T suppression. Collectively, these data indicate that 4-1BB-dependent suppression of SEA specific CD4(+) T responses was mediated mainly by IFN-gamma-dependent IDO induction and partially by TGF-beta. PMID- 19218485 TI - Beam damage suppression of low-kappa porous Si-O-C films by cryo-electron-energy loss spectroscopy (EELS). AB - Porous Si-O-C films with lower dielectric constant (kappa) relative to silicon dioxide have been widely studied as inter-layer dielectrics in new-generation microelectronic devices. On the analysis of the film by transmission electron microscopy (TEM), it is susceptible of beam damage during both sample preparation by a focused ion beam (FIB) technique and TEM observation. We use electron energy loss spectroscopy (EELS) to quantify the magnitude of the beam damage during these processes. The intensity of the 285-eV peak in C-K electron energy loss near edge structures (ELNES) is enhanced by the damage, which can be ascribed to the formation of the C=C double bonds as a result of the decomposition of the methyl groups by the beam. The use of cryo-holder for TEM at 100 K is found to be essential to reduce the damage of the low-kappa layers. The lowering of the acceleration voltage of FIB down to 5 keV does not change the spectra. Since the FIB damage is localized at the surface, the use of thick regions in the TEM foil such as 130 nm is preferred to reduce the superposition of EELS of the damaged region on those from the sample of interest. PMID- 19218486 TI - Visualization of the ultrastructural interface of cells with the outer and inner surface of coral skeletons. AB - Crystalline, porous biomaterials, such as marine invertebrate skeletons, have been widely used for functional reconstruction of human tissues like bone and dental implants. Since in such an abrasive microenvironment adequate cell material interactions are crucial for a successful treatment, it is of great importance to improve the means to examine these interactions. We developed a method that reveals the ultrastructure of the interface between coral skeletons and cultured neural cells to a higher quality than do traditional methods as it does not include damaging procedures like decalcification or sectioning non decalcified skeletons. It is rather based on generating two electron opacity distinct Araldite masks, of the skeleton and its surrounding, by polymerizing them to different durations. The contrast created at the border of the two masks outlined the fine and fragile crystals of the coral skeleton's outer and inner surfaces and their contact sites with the cells. The skeleton's internal structure contains a mesh of narrow (few microns wide) and large channel-shaped gaps interrupted by irregular-shaped crystalline material. Neural cells grew on the skeleton surface by stretching between crystal tips, with occasional rearrangements of cytoskeletal fibers located near the anchorage focal adherence points. Cell processes infiltrated the skeleton interior by stretching between inter-surface crystals and by adjusting their volume to the space of the conduits they grew into. The technique advances the study of coral biology and of neural cells-hard biomaterial interaction; it can be applied to other biomaterials and cell types and open new ways for studying tissue development and engineering. PMID- 19218487 TI - Oral administration of pharmacologically active substances to squid: a methodological description. AB - The squid giant synapse is a well-defined experimental preparation for the study of ligand-dependant synaptic transmission. Its large size gives direct experimental access to both presynaptic and postsynaptic junctional elements, allowing direct optical, biophysical, and electrophysiological analysis of depolarization-release coupling. However, this important model has not been utilized in pharmacological studies, other than those implementable acutely in the in vitro condition. A method is presented for oral administration of bioactive substances to living squid. Electrophysiological characterization and direct determination of drug absorption into the nervous system demonstrate the administration method described here to be appropriate for pharmacological research. PMID- 19218488 TI - Behaving in the dark: locomotor, chromatic, postural, and bioluminescent behaviors of the deep-sea squid Octopoteuthis deletron young 1972. AB - Visual behaviors are prominent components of intra- and interspecific communication in shallow-water cephalopods. Meso- and bathypelagic cephalopods were believed to have limited visual communication, other than bioluminescence, due to the reduced illumination at depth. To explore potential visual behaviors in mesopelagic squid, we used undersea vehicles to observe 76 individuals of Octopoteuthis deletron. In contrast to predictions, we found this species capable of a variety of visually linked behaviors not previously reported for a deep ocean cephalopod. The resultant ethogram describes numerous chromatic, postural, locomotor, and bioluminescent behavioral components. A few common body patterns the whole appearance of the individual involving multiple components-are characterized. The behaviors observed from individual squid were compared using a Non-metric Multi-Dimensional Scaling (NMDS) ordination, onto which hydrographic and observation parameters were mapped. Observation length, specimen collection, and contact with the vehicle affected which behaviors were performed. A separate NMDS, analyzing the body patterns, indicated that these sets of behavioral components could be visualized as groups within the NMDS ordination. While the functional roles of the behaviors described are not yet known, our findings of numerous behaviors in O. deletron clearly indicate that bioluminescence is not the sole method of visual communication by deep-sea squid. PMID- 19218489 TI - Mass culture and characterization of tumor cells from a naturally occurring invertebrate cancer model: applications for human and animal disease and environmental health. AB - On the northeastern coast of the United States and Canada, Mya arenaria, the soft shell clam, develops a diffuse, hemopoetic tumor (a fatal leukemia-like cancer) resulting from inactivation of p53-like family member proteins.These malignant cells provide a model for an unrelated set of human cancer cells that are also characterized by mortalin-based cytoplasmic sequestration of wild-type p53 tumor suppressor protein (mortalin is the mitochondrial Hsp70 protein). Here we describe methods for mass culture and long-term storage of tumor cells from this cancer. These are the first successful efforts at maintaining malignant cells from any marine invertebrate in vitro. Following passage (subculture), these cultures undergo transition from primary cultures to non-immortalized cell lines that continue to proliferate and do not re-differentiate the normal hemocyte phenotype. We also characterize normal clam hemocytes and the pathology of cancerous clam hemocytes in vitro and in vivo using light and electron microscopy, cyto- and immunocytochemistry, molecular biology, and a phagocytosis assay. Our protocols provide biomedical and environmental researchers with ready access to this naturally occurring cancer model. We discuss the clam cancer model regarding (a) human health and disease; (b) animal health, disease, and aquaculture; (c) environmental health monitoring; and (d) future research directions. PMID- 19218490 TI - Calyculin-A induces cleavage in a random plane in unfertilized sea urchin eggs. AB - Calyculin-A (CLA), a protein phosphatase inhibitor, has been known to induce cleavage resembling normal furrowing in unfertilized sea urchin eggs. In CLA treated eggs, actin filaments and myosin assemble to form a contractile ring-like structure in the egg cortex; however, this occurs in the absence of a mitotic spindle or asters. Here, we investigated the relationship between the plane of CLA-induced cleavage and the intrinsic animal-vegetal polar axis in sea urchin eggs. The animal-vegetal axis was established using black ink to visualize the jelly canal located at the animal pole in the jelly coat surrounding the egg. We measured the acute angle between the jelly canal axis and the cleavage plane for both fertilized eggs and CLA-treated unfertilized eggs. Although the acute angle lay within 10 degrees for most of the fertilized eggs, it varied widely for CLA treated unfertilized eggs. Measurements of the diameter of blastomeres revealed that cleavage of fertilized eggs took place in the mid-plane of the egg, but that CLA-induced divisions were unequal. These results suggest that neither the orientation nor the location of the CLA-induced cleavage furrow is related to the animal-vegetal polar axis of the egg, even though the furrowing mechanism itself is not dissimilar to that in fertilized eggs. PMID- 19218491 TI - Low oxygen consumption and high body content of catch connective tissue contribute to low metabolic rate of sea cucumbers. AB - The energy consumption of echinoderms is low in comparison with that of other invertebrates. We demonstrated this by measuring the oxygen consumption rate per unit of body weight (VO2) of the sea cucumber Actinopyga mauritiana: VO2 was 1/8 that of the "standard" invertebrates. Low energy consumption in echinoderms has been attributed to their high skeletal content and to catch connective tissues (CCTs) that maintain body posture by altering their mechanical properties with little energy expenditure. The former is not applicable to holothurians, and the latter has not been proven experimentally. We postulated that the large content of dermal connective tissue, which maintaines posture economically, contributes to the low energy consumption in holothurians. Body-wall dermis occupied 53.5% of wet body weight, whereas body-wall muscles, including those of tube feet, occupied 5.1%. VO2 of the dermis in the stiff state (2.45 microl x g(-1) x h(-1)) was 1/10 that of the longitudinal body-wall muscle in contraction. the mechanical tests revealed that the stress at an imposed strain of 2% strain was 7 times greater in CCT than in muscles. These results showed that CCT could maintain posture more economically than muscles could. We concluded that the high content of connective tissue with energy-saving posture-maintenance activities contributed to the low energy consumption of holothurians. PMID- 19218492 TI - Temporal variability in chlorophyll fluorescence of back-reef corals in Ofu, American Samoa. AB - Change in the yield of chlorophyll a fluorescence is a common indicator of thermal stress in corals. The present study reports temporal variability in quantum yield measurements for 10 coral species in Ofu, American Samoa-a place known to experience elevated and variable seawater temperatures. In winter, the zooxanthellae generally had higher dark-adapted maximum quantum yield (F(v)/F(m)), higher light-adapted effective quantum yield (DeltaF/F'(m)), and lower relative electron transport rates (rETR) than in the summer. Temporal changes appeared unrelated to the expected bleaching sensitivity of corals. All species surveyed, with the exception of Montipora grisea, demonstrated significant temporal changes in the three fluorescence parameters. Fluorescence responses were influenced by the microhabitat-temporal differences in fluorescence parameters were usually observed in the habitat with a more variable temperature regime (pool 300), while differences in F(v)/F(m) between species were observed only in the more environmentally stable habitat (pool 400). Such species-specific responses and microhabitat variability should be considered when attempting to determine whether observed in situ changes are normal seasonal changes or early signs of bleaching. PMID- 19218493 TI - Red fluorescent protein responsible for pigmentation in trematode-infected Porites compressa tissues. AB - Reports of coral disease have increased dramatically over the last decade; however, the biological mechanisms that corals utilize to limit infection and resist disease remain poorly understood. Compromised coral tissues often display non-normal pigmentation that potentially represents an inflammation-like response, although these pigments remain uncharacterized. Using spectral emission analysis and cryo-histological and electrophoretic techniques, we investigated the pink pigmentation associated with trematodiasis, infection with Podocotyloides stenometre larval trematode, in Porites compressa. Spectral emission analysis reveals that macroscopic areas of pink pigmentation fluoresce under blue light excitation (450 nm) and produce a broad emission peak at 590 nm (+/-6) with a 60-nm full width at half maximum. Electrophoretic protein separation of pigmented tissue extract confirms the red fluorescence to be a protein rather than a low-molecular-weight compound. Histological sections demonstrate green fluorescence in healthy coral tissue and red fluorescence in the trematodiasis-compromised tissue. The red fluorescent protein (FP) is limited to the epidermis, is not associated with cells or granules, and appears unstructured. These data collectively suggest that the red FP is produced and localized in tissue infected by larval trematodes and plays a role in the immune response in corals. PMID- 19218494 TI - Seawater temperature alters feeding discrimination by cold-temperate but not subtropical individuals of an ectothermic herbivore. AB - Seawater temperature varies across multiple spatial and temporal scales, yet the roles that such variation play in altering biotic interactions are poorly known. We assessed temperature-mediated feeding behavior exhibited by the herbivorous amphipod Ampithoe longimana collected from cold-temperate and subtropical estuaries (27 degrees N and 41 degrees N, respectively). Individuals were offered a pairwise feeding choice between lyophilized seaweeds that provide higher fitness (Ulva intestinalis) or lower fitness (Halimeda tuna, H. opuntia, Amphiroa spp., or Stypopodium zonale). Overall, herbivores preferentially consumed the higher quality U. intestinalis more than any lower quality food. However, the strength of this feeding choice was not consistent. Northern herbivores consumed proportionally more poorer quality tissue at 25 degrees C than at 20 degrees C in two assays (H. opuntia and Amphiroa sp.), consumed less poorer quality tissue at 25 degrees C than at 20 degrees C in one assay (S. zonale), and showed no difference in another assay (H. tuna). Moreover, when offered tissue coated with lipophilic extracts of H. opuntia, northern herbivores consumed more extract coated tissue at 25 degrees C than at 20 degrees C. In contrast to northern herbivores, the southern herbivores did not alter their feeding choices with temperature. This study represents the first demonstration that short-term (i.e., days-long) variation in ambient temperature alters feeding choices in a marine herbivore. PMID- 19218495 TI - Diel variation in the sizes of larvae of Bugula neritina in field populations. AB - The scale of planktonic larval dispersal affects a variety of ecological and evolutionary processes. Recent work suggests that the dispersal ability of obligately lecithotrophic larvae is influenced by the amount of energy supplied to each larva: larger larvae may stay in the plankton longer and thus travel greater distances than smaller larvae. We examined a prediction of this hypothesis in the bryozoan Bugula neritina, which each morning releases brooded larvae that settle within a few hours. If larger larvae stay in the plankton longer than smaller larvae, than larger larvae should increase in frequency in the planktonic population as the day progresses. However, field surveys revealed a negative relationship between time of day and the sizes of planktonic larvae. Because these results may have been complicated by prolonged larval release, we sequestered groups of brooding colonies in field mesocosms to examine release patterns. Larvae were released over a period of 8-9 h, with smaller larvae increasing in frequency as the day progressed. We conclude that populations of larvae of B. neritina may not be homogenous in energetic content throughout the day; this must be taken into consideration when designing studies of many aspects of larval biology. PMID- 19218496 TI - Gonad development during the early life of Octopus maya (Mollusca: Cephalopoda). AB - Gonad development during the early life of Octopus maya is described in terms of histological, morphometric, oocytes growth, and somatic-oocyte relationship data obtained from octopus cultured at the UMDI-UNAM, in Sisal, Yucatan, Mexico. This study is the first publication on gonad development during the early life of Octopus maya. A total of 83 O. maya specimens were used; their sizes ranged from 6.5 to 76 mm of total length (TL), 4 to 28 mm of dorsal mantle length (DML), 2.5 to 20 mm of ventral mantle length (VML), and 0.0180 to 7.2940 g of fixed body weight (fBW). Animals were weighed and measured only after preservation. A loss of 10% of living weight was estimated for juvenile octopuses after formalin preservation. The relation of length to weight (VML, DML, TL/fBW) pooled for both sexes had a strong positive correlation (r), as shown by a potential power function that was quite close to 1. Compound images were produced from numerous microscopic fields. The histological examination revealed that, 4 months after hatching, male octopus (24.5 mm DML and 7.2940 g fBW) were in gonad stages 2 (maturing) to 3 (mature), with spermatogonia and spermatocytes in the tubule wall and abundant spermatids and spermatozoa in the central lumen of the seminiferous tubules, suggesting the occurrence of different phases of gonad development at different maturity stages. In contrast, females (22.5 mm DML and 4.8210 g fBW) at the same time since hatching were immature (stage 1), with many oogonia, few oocytes, and germinal epithelium. This suggests that males reach maturity earlier than females, indicating a probable onset of maturity for males at around 4 months of culture or 8 g of wet body weight. Our results indicate the possibility that the size-at-weight can be recognized early with a degree of certainty that allows the sexes to be separated for culture purposes; but more detailed studies on reproduction in relation to endocrinology and nutrition are needed. PMID- 19218498 TI - Induction of macroautophagy by overexpression of the Parkinson's disease associated GPR37 receptor. AB - The orphan G-protein-coupled receptor 37 (GPR37) is a substrate of parkin, and its insoluble aggregates accumulate in brain tissue samples of Parkinson's disease patients, including Lewy bodies and neurites. Parkin activates the clearance of the unfolded receptor, while the overexpression of GPR37, in the absence of parkin, can lead to unfolded protein-induced cell death. We found that overexpression of the human GPR37 receptor in HEK293 cells and consequent activation of an endoplasmic reticulum (ER) stress response had effects comparable to starvation, in inducing the cellular autophagic pathway. Treatment with specific modulators provided further evidence for the autophagic clearance of the overexpressed GPR37 protein, in detergent-soluble and -insoluble fractions, as confirmed by the conversion of the microtubule-associated protein 1, light chain 3 (LC3)-I marker to its LC3-II isoform. Furthermore, Gpr37-null mutant mice displayed consistent alterations of ER stress and autophagic pathway markers in brain tissue samples. These findings show that GPR37 overexpression per se can induce cellular autophagy, which may prevent the selective degeneration of GPR37-expressing neurons, as reported for Parkinson's and related neurodegenerative diseases. PMID- 19218497 TI - Astaxanthin reduces ischemic brain injury in adult rats. AB - Astaxanthin (ATX) is a dietary carotenoid of crustaceans and fish that contributes to their coloration. Dietary ATX is important for development and survival of salmonids and crustaceans and has been shown to reduce cardiac ischemic injury in rodents. The purpose of this study was to examine whether ATX can protect against ischemic injury in the mammalian brain. Adult rats were injected intracerebroventricularly with ATX or vehicle prior to a 60-min middle cerebral artery occlusion (MCAo). ATX was present in the infarction area at 70-75 min after onset of MCAo. Treatment with ATX, compared to vehicle, increased locomotor activity in stroke rats and reduced cerebral infarction at 2 d after MCAo. To evaluate the protective mechanisms of ATX against stroke, brain tissues were assayed for free radical damage, apoptosis, and excitoxicity. ATX antagonized ischemia-mediated loss of aconitase activity and reduced glutamate release, lipid peroxidation, translocation of cytochrome c, and TUNEL labeling in the ischemic cortex. ATX did not alter physiological parameters, such as body temperature, brain temperature, cerebral blood flow, blood gases, blood pressure, and pH. Collectively, our data suggest that ATX can reduce ischemia-related injury in brain tissue through the inhibition of oxidative stress, reduction of glutamate release, and antiapoptosis. ATX may be clinically useful for patients vulnerable or prone to ischemic events. PMID- 19218499 TI - Functional characterization of a promoter polymorphism that drives ACSL5 gene expression in skeletal muscle and associates with diet-induced weight loss. AB - Diet-induced weight loss is affected by a wide range of factors, including genetic variation. Identifying functional polymorphisms will help to elucidate mechanisms that account for variation in dietary metabolism. Previously, we reported a strong association between a common single nucleotide polymorphism (SNP) rs2419621 (C>T) in the promoter of acyl-CoA synthetase long chain 5 (ACSL5), rapid weight loss in obese Caucasian females, and elevated ACSL5 mRNA levels in skeletal muscle biopsies. Here, we showed by electrophoretic mobility shift assay (EMSA) that the T allele creates a functional cis-regulatory E-box element (CANNTG) that is recognized by the myogenic regulatory factor MyoD. The T allele promoted MyoD-dependent activation of a 1089-base pair ACSL5 promoter fragment in nonmuscle CV1 cells. Differentiation of skeletal myoblasts significantly elevated expression of the ACSL5 promoter. The T allele sustained promoter activity 48 h after differentiation, whereas the C allele showed a significant decline. These results reveal a mechanism for elevated transcription of ACSL5 in skeletal muscle of carriers of the rs2419621(T) allele, associated with more rapid diet-induced weight loss. Natural selection favoring promoter polymorphisms that reduced expression of catabolic genes in skeletal muscle likely accounts for the resistance of obese individuals to dietary intervention. PMID- 19218500 TI - Endothelial heparanase secretion after acute hypoinsulinemia is regulated by glucose and fatty acid. AB - Following diabetes, the heart increases its lipoprotein lipase (LPL) at the coronary lumen by transferring LPL from the cardiomyocyte to the endothelial lumen. We examined how hyperglycemia controls secretion of heparanase, the enzyme that cleaves myocyte heparan sulphate proteoglycan to initiate this movement. Diazoxide (DZ) was used to decrease serum insulin and generate hyperglycemia. A modified Langendorff technique was used to separate coronary from interstitial effluent, which were assayed for heparanase and LPL. Within 30 min of DZ, interstitial heparanase increased, an effect that closely mirrored an augmentation in interstitial LPL. Endothelial cells were incubated with palmitic acid (PA) or glucose, and heparanase secretion was determined. PA increased intracellular heparanase, with no effect on secretion of this enzyme. Unlike PA, glucose dose-dependently lowered endothelial intracellular heparanase, which was strongly associated with increased heparanase activity in the incubation medium. Preincubation with cytochalasin D or nocodazole prevented the high glucose induced depletion of intracellular heparanase. Our data suggest that following hyperglycemia, translocation of LPL from the cardiomyocyte cell surface to the apical side of endothelial cells is dependent on the ability of the fatty acid to increase endothelial intracellular heparanase followed by rapid secretion of this enzyme by glucose, which requires an intact microtubule and actin cytoskeleton. PMID- 19218501 TI - Advanced methods for quantification of infarct size in mice using three dimensional high-field late gadolinium enhancement MRI. AB - Conventional methods to quantify infarct size after myocardial infarction in mice are not ideal, requiring either tissue destruction for histology or relying on nondirect measurements such as wall motion. We therefore implemented a fast, high resolution method to directly measure infarct size in vivo using three dimensional (3D) late gadolinium enhancement MRI (3D-LGE). Myocardial T1 relaxation was quantified at 9.4 Tesla in five mice, and reproducibility was tested by repeat imaging after 5 days. In a separate set of healthy and infarcted mice (n = 8 of each), continuous T1 measurements were made following intravenous or intraperitoneal injection of a contrast agent (0.5 micromol/g gadolinium diethylenetriamine pentaacetic acid). The time course of T1 contrast development between viable and nonviable myocardium was thereby determined, with optimal postinjection imaging windows and inversion times identified. Infarct sizes were quantified using 3D-LGE and compared with triphenyltetrazolium chloride histology on day 1 after infarction (n = 8). Baseline myocardial T1 was highly reproducible: the mean value was 952 +/- 41 ms. T1 contrast peaked earlier after intravenous injection than with intraperitoneal injection; however, contrast between viable and nonviable myocardium was comparable for both routes (P = 0.31), with adequate contrast remaining for at least 60 min postinjection. Excellent correlation was obtained between infarct sizes derived from 3D-LGE and histology (r = 0.91, P = 0.002), and Bland-Altman analysis indicated good agreement free from systematic bias. We have validated an improved 3D MRI method to noninvasively quantify infarct size in mice with unsurpassed spatial resolution and tissue contrast. This method is particularly suited to studies requiring early quantification of initial infarct size, for example, to measure damage before intervention with stem cells. PMID- 19218502 TI - Rho-kinase-mediated suppression of KDR current in cerebral arteries requires an intact actin cytoskeleton. AB - This study examined the role of the actin cytoskeleton in Rho-kinase-mediated suppression of the delayed-rectifier K(+) (K(DR)) current in cerebral arteries. Myocytes from rat cerebral arteries were enzymatically isolated, and whole cell K(DR) currents were monitored using conventional patch-clamp electrophysiology. At +40 mV, the K(DR) current averaged 19.8 +/- 1.6 pA/pF (mean +/- SE) and was potently inhibited by UTP (3 x 10(-5) M). This suppression was observed to depend on Rho signaling and was abolished by the Rho-kinase inhibitors H-1152 (3 x 10( 7) M) and Y-27632 (3 x 10(-5) M). Rho-kinase was also found to concomitantly facilitate actin polymerization in response to UTP. We therefore examined whether actin dynamics played a role in the ability of Rho-kinase to suppress K(DR) current and found that actin disruption using either cytochalasin D (1 x 10(-5) M) or latrunculin A (1 x 10(-8) M) prevented current modulation. Consistent with our electrophysiological observations, both Rho-kinase inhibition and actin disruption significantly attenuated UTP-induced depolarization and constriction of cerebral arteries. We propose that UTP initiates Rho-kinase-mediated remodeling of the actin cytoskeleton and consequently suppresses the K(DR) current, thereby facilitating the depolarization and constriction of cerebral arteries. PMID- 19218503 TI - Differential regulation of angiotensin-(1-12) in plasma and cardiac tissue in response to bilateral nephrectomy. AB - We examined the effects of 48 h bilateral nephrectomy on plasma and cardiac tissue expression of angiotensin-(1-12) [ANG-(1-12)], ANG I, and ANG II in adult Wistar-Kyoto rats to evaluate functional changes induced by removing renal renin. The goal was to expand the evidence of ANG-(1-12) being an alternate renin independent, angiotensin-forming substrate. Nephrectomy yielded divergent effects on circulating and cardiac angiotensins. Significant decreases in plasma ANG-(1 12), ANG I, and ANG II levels postnephrectomy accompanied increases in cardiac ANG-(1-12), ANG I, and ANG II concentrations compared with controls. Plasma ANG (1-12) decreased 34% following nephrectomy, which accompanied 78 and 66% decreases in plasma ANG I and ANG II, respectively (P < 0.05 vs. controls). Contrastingly, cardiac ANG-(1-12) in anephric rats averaged 276 +/- 24 fmol/mg compared with 144 +/- 20 fmol/mg in controls (P < 0.005). Cardiac ANG I and ANG II values were 300 +/- 15 and 62 +/- 7 fmol/mg, respectively, in anephric rats compared with 172 +/- 8 fmol/mg for ANG I and 42 +/- 4 fmol/mg for ANG II in controls (P < 0.001). Quantitative immunofluorescence revealed significant increases in average grayscale density for cardiac tissue angiotensinogen, ANG I, ANG II, and AT(1) receptors of WKY rats postnephrectomy. Faint staining of cardiac renin, unchanged by nephrectomy, was associated with an 80% decrease in cardiac renin mRNA. These changes were accompanied by significant increases in p47(phox), Rac1, and Nox4 isoform expression. In conclusion, ANG-(1-12) may be a functional precursor for angiotensin peptide formation in the absence of circulating renin. PMID- 19218504 TI - Cardiac systolic and diastolic function during whole body heat stress. AB - During a whole body heat stress, stroke volume is either maintained or slightly elevated despite reduced ventricular filling pressures and central blood volume, suggestive of improved cardiac diastolic and/or systolic function. Heat stress improves cardiac systolic and diastolic function in patients with congestive heart failure, although it remains unknown whether similar responses occur in healthy individuals, which is the hypothesis to be tested. Nine male volunteers underwent a whole body heat stress. Echocardiographic indexes of diastolic and systolic function were performed following a supine resting period, and again following an increase in internal temperature of approximately 1.0 degrees C via passive heat stress. Despite previous reports of heat stress-induced decreases in ventricular filling pressures and central blood volume, no changes in indexes of diastolic function were identified during heating [i.e., unchanged early diastolic mitral annular tissue velocity (E'), mitral inflow during the early diastolic phase (E), the E/E' ratio, and isovolumetric relaxation time]. Heat stress increased late diastolic septal (P = 0.03) and lateral (P = 0.01) mitral annular tissue velocities (A'), mitral inflow velocity during atrial contraction (P < 0.001), and the relative contribution of atrial contraction to left ventricular filling during diastole (P = 0.01), all indicative of improved atrial systolic function. Furthermore, indexes of ventricular systolic function were increased by heat stress [i.e., increased septal (P = 0.001) and lateral (P = 0.01) mitral annular systolic velocities and isovolumic acceleration at the septal (P = 0.03) and lateral (P < 0.001) mitral annulus]. These data are suggestive of improved atrial and ventricular systolic function by the heat stress. Together these data support previous findings, which used the less precise measure of ejection fraction, that heat stress improves indexes of systolic function, while diastolic function is maintained. PMID- 19218505 TI - Translocon closure to Ca2+ leak in proliferating vascular smooth muscle cells. AB - Vascular smooth muscle cells have a proliferative phenotype that is important in vascular development, adaptation, and disease. Intracellular calcium handling is thought to play pivotal roles in determining the properties of these cells, and thus previously unrecognized mechanisms for transmembrane calcium movement are of potential interest. An unsolved question is the mechanism of constitutive (passive) calcium leak from the intracellular stores. Studies of other cell types have suggested that the translocon is a calcium leak pathway. Here we investigated the contribution of the translocon in proliferating vascular smooth muscle cells. Calcium leak into the cytoplasm was measured using fura-2, and protein synthesis was measured using radioactive methionine. Puromycin, emetine, and anisomycin are chemicals that inhibit protein synthesis, acting via the translocon; all three agents strongly inhibited protein synthesis in the smooth muscle cells within 1 h. Puromycin, which opens the translocon, evoked a transient increase in cytoplasmic calcium that was similar in amplitude to the calcium rise evoked by thapsigargin. The puromycin effect was abolished by thapsigargin. The treatment of cells for 1 h with emetine or anisomycin, which close the translocon, inhibited the calcium release evoked by puromycin but not the calcium release evoked by extracellular ATP, endothelin-1, or the calcium ionophore ionomycin. Thapsigargin-evoked calcium rises were slightly suppressed by emetine but unaffected by puromycin or anisomycin. The data suggest that the translocon has the capacity to act as a calcium leak pathway in the ribosomal endoplasmic reticulum but that it is normally closed and lacks relevance to physiological calcium leak mechanisms. PMID- 19218506 TI - Physiological roles of A1 and A2A adenosine receptors in regulating heart rate, body temperature, and locomotion as revealed using knockout mice and caffeine. AB - Heart rate (HR), body temperature (Temp), locomotor activity (LA), and oxygen consumption (O(2)C) were studied in awake mice lacking one or both of the adenosine A(1) or A(2A) receptors (A(1)R or A(2A)R, respectively) using telemetry and respirometry, before and after caffeine administration. All parameters were lower during day than night and higher in females than males. When compared with wild-type (WT) littermates, HR was higher in male A(1)R knockout (A(1)RKO) mice but lower in A(2A)RKO mice and intermediate in A(1)-A(2A)R double KO mice. A single dose of an unselective beta-blocker (timolol; 1 mg/kg) abolished the HR differences between these genotypes. Deletion of A(1)Rs had little effect on Temp, whereas deletion of A(2A)Rs increased it in females and decreased it in males. A(1)-A(2A)RKO mice had lower Temp than WT mice. LA was unaltered in A(1)RKO mice and lower in A(2A)RKO and A(1)-A(2A)RKO mice than in WT mice. Caffeine injection increased LA but only in mice expressing A(2A)R. Caffeine ingestion also increased LA in an A(2A)R-dependent manner in male mice. Caffeine ingestion significantly increased O(2)C in WT mice, but less in the different KO mice. Injection of 30 mg/kg caffeine decreased Temp, especially in KO mice, and hence in a manner unrelated to A(1)R or A(2A)R blockade. Selective A(2B) antagonism had little or no effect. Thus A(1)R and A(2A)R influence HR, Temp, LA, and O(2)C in mice in a sex-dependent manner, indicating effects of endogenous adenosine. The A(2A)R plays an important role in the modulation of O(2)C and LA by acute and chronic caffeine administration. There is also evidence for effects of higher doses of caffeine being independent of both A(1)R and A(2A)R. PMID- 19218507 TI - Role of ROCK I/II in vascular branching. PMID- 19218508 TI - The physiology of long-distance migration: extending the limits of endurance metabolism. AB - Long-distance migrants have evolved specific adaptations that make their athletic records possible. Unique mechanisms explaining their amazing capacity for endurance exercise have now been uncovered, particularly with respect to energy storage, mobilization, transport and utilization. Birds are champions of migration because flying offers a key compromise: it allows more rapid movement than swimming, but has a lower cost of transport than running. High efficiency for muscle contraction, pointed wings, low wingloading, travelling in V formations, storing fuel as energy-dense lipids and atrophy of non-essential organs are some of their strategies to decrease the cost of transport. The ability to process lipids rapidly also emerges as a crucial component of the migrant phenotype. High lipid fluxes are made possible by lipoprotein shuttles and fatty acid binding proteins (FABPs) that accelerate lipid transport and by upgrading the metabolic machinery for lipolysis and lipid oxidation. Preparation for long flights can include natural doping on n-3 polyunsaturated fatty acids (n 3 PUFAs) from unique invertebrate diets. Muscle performance is improved by restructuring membrane phospholipids and by activating key genes of lipid metabolism through peroxisome proliferator-activated receptors (PPARs). The physiological secret to long migrations does not depend on a single ;magic' adaptation but on the integration of multiple adjustments in morphology, biomechanics, behavior, nutrition and metabolism. Research on the physiology of migrants improves the fundamental knowledge of exercise biology, but it also has important implications for wildlife conservation, treating obesity and improving the performance of human athletes. PMID- 19218509 TI - Muscular and non-muscular contributions to maximum power cycling in children and adults: implications for developmental motor control. AB - During submaximal cycling, children demonstrate a different distribution between muscular and non-muscular (gravitational and motion-dependent) forces when compared with adults. This is partly due to anthropometric differences. In this study, we tested the hypothesis that during maximum power cycling, children would construct the task (in terms of the distribution between muscular and non muscular pedal power) similarly to adults. Eleven children (aged 8-9 years) and 13 adults (aged 20-40 years) performed a maximal isokinetic cycling task over 3 s at 115 r.p.m. Multivariate analyses of variance revealed no significant differences in normalized maximum, minimum and average positive non-muscular pedal power between children and adults (Wilks' lambda=0.755, F(3,20)=2.17, P=0.124). Thus, maximum cycling is a developmental ;self-scaling' task and age related differences in muscular power production are not confounded by differences in anthropometry. This information is useful to researchers who wish to differentiate between muscular and non-muscular power when studying developmental motor control. In addition to the similarities in the distribution between muscular and non-muscular pedal power, we found age-related differences in the relative joint power contributions to total pedal power. In children, a significantly smaller proportion of total pedal power was generated at the ankle joint (6.1+/-5.4% for children and 12.6+/-3.2% for adults), whilst relatively more power was generated at the knee and hip joints. These results suggest that intermuscular coordination may be contributing to children's limits in maximum power production during multi-joint tasks. PMID- 19218510 TI - Losing stability: tail loss and jumping in the arboreal lizard Anolis carolinensis. AB - Voluntary loss of an appendage, or autotomy, is a remarkable behavior that is widespread among many arthropods and lower vertebrates. Its immediate benefit, generally escape from a predator, is balanced by various costs, including impaired locomotor performance, reproductive success and long-term survival. Among vertebrates, autotomy is most widespread in lizards, in which tail loss has been documented in close to 100 species. Despite numerous studies of the potential costs of tail autotomy in lizards, none have focused on the importance of the tail in jumping. Using high-speed video we recorded jumps from six lizards (Anolis carolinensis) both before and after removing 80% of the tail to test the hypothesis that tail loss has a significant effect on jumping kinematics. Several key performance metrics, including jump distance and takeoff velocity, were not affected by experimental tail removal, averaging 21 cm and 124 cm s(-1), respectively, in both tailed and tailless lizards. However, in-air stability during jumping was greatly compromised after tail removal. Lizards without tails rotated posteriorly more than 30 deg., on average, between takeoff and landing (and sometimes more than 90 deg.) compared with an average of 5 deg. of rotation in lizards with intact tails. Such exaggerated posterior rotation prevents coordinated landing, which is critical for animals that spend much of their time jumping to and from small branches. This work augments recent experiments demonstrating the importance of the tail as a mid-air stabilizer during falling in geckos, and emphasizes new and severe functional costs associated with tail autotomy in arboreal lizards. PMID- 19218511 TI - Aerobic capacity and running performance across a 1.6 km altitude difference in two sciurid rodents. AB - Hypoxia at high altitudes is often assumed to constrain exercise capacity, but there have been few high- versus low-altitude comparisons of species native to a wide range of altitudes. Such studies are ecologically realistic, as wild-caught animals tested at their native altitude are presumably maximally acclimated (via phenotypic plasticity) or adapted (by evolutionary change) to that altitude. We compared aerobic performance, measured as maximum oxygen consumption in forced exercise (V(O(2),max)), and voluntary wheel-running in two species of sciurid rodents captured and tested at field sites that differed in altitude by 1.6 km (2165 m versus 3800 m). We found reduced V(O(2),max) at 3800 m in least chipmunks (Tamias minimus) but no significant effect of altitude on V(O(2),max) in golden mantled ground squirrels (Spermophilus lateralis). Individuals of both species averaged several km day(-1) in wheels. Most behavioral indices of voluntary running (including mean and maximum speeds, time spent running, daily running distance, and the number and duration of running bouts) were unaffected by altitude, even in the species with reduced V(O(2),max) at high altitude. Metabolic rates during running and energy costs of transport differed to some extent across altitudes but in different ways in the two species. At both test sites, voluntary running by both species was almost exclusively at speeds well within aerobic limits. We conclude that substantial differences in altitude do not necessarily result in differences in aerobic capacity in small mammals and, even if V(O(2),max) is reduced at high altitude, there may be no effect on voluntary running behavior. PMID- 19218512 TI - Odour aversion after olfactory conditioning of the sting extension reflex in honeybees. AB - In Pavlovian conditioning, an originally neutral stimulus (conditioned stimulus or CS) gains control over an animal's reflex after its association with a biologically relevant stimulus (unconditioned stimulus or US). As a consequence, a conditioned response is emitted by the animal upon further CS presentations. In such a situation, the subject exhibits a reflex response, so that whether the CS thereby acquires a positive or a negative value for the animal is difficult to assess. In honeybees, Apis mellifera, an odour (CS) can be associated either with sucrose solution (US) in the appetitive conditioning of the proboscis extension reflex (PER), or with an electric shock (US) in the aversive conditioning of the sting extension reflex (SER). The term ;aversive' may not apply to the latter as bees do not suppress SER as a consequence of learning but, on the contrary, start emitting SER to the CS. To determine whether the CS acquires a positive or a negative value in these conditioning forms, we compared the orientation behaviour of freely walking honeybees in an olfactory-cued Y-maze after training them with an odour-sucrose association (PER conditioning) or an odour-shock association (SER conditioning). We show that the same odours can acquire either a positive value when associated to sucrose, or a negative value when associated to an electric shock, as bees respectively approach or avoid the CS in the Y-maze. Importantly, these results clearly establish the aversive nature of SER conditioning in honeybees. PMID- 19218513 TI - Diverse cell-specific expression of myoglobin isoforms in brain, kidney, gill and liver of the hypoxia-tolerant carp and zebrafish. AB - Myoglobin (Mb) is famous as a muscle-specific protein--yet the common carp expresses the gene (cMb1) encoding this protein in a range of non-muscle tissues and also expresses a novel isoform (cMb2) in the brain. Using a homologous antibody and riboprobes, we have established the relative amounts and cellular sites of non-muscle Mb expression in different tissues. The amounts of carp myoglobin (cMb) in supernatants of different tissues were just 0.4-0.7% relative to that of heart supernatants and were upregulated by two-to-four fold in liver, gill and brain following 5 days of hypoxic treatment. Brain exhibited both cMb proteins in western analysis, whereas all other tissues had only cMb1. We have also identified cells expressing cMb protein and cMb mRNA using immunohistology and RNA in situ hybridisation (RNA-ISH), respectively. Mb was strongly expressed throughout all cardiac myocytes and a subset of skeletal muscle fibres, whereas it was restricted to a small range of specific cell types in each of the non muscle tissues. These include pillar and epithelial cells in secondary gill lamellae, hepatocytes, some neurones, and tubular epithelial cells in the kidney. Capillaries and small blood vessels in all tissues exhibited Mb expression within vascular endothelial cells. The cMb2 riboprobe located expression to a subset of neurones but not to endothelial cells. In zebrafish, which possesses only one Mb gene, a similar expression pattern of Mb protein and mRNA was observed. This establishes a surprisingly cell-specific distribution of Mb within non-muscle tissues in both carp and zebrafish, where it probably plays an important role in the regulation of microvascular, renal and brain function. PMID- 19218514 TI - Plasma membrane calcium ATPase required for semicircular canal formation and otolith growth in the zebrafish inner ear. AB - Fish otoliths consist of >90% calcium carbonate, the accretion of which depends on acellular endolymph. This study confirms the presence of plasma membrane calcium ATPase 1a isoform (Atp2b1a) in the auditory and vestibular system of a teleost fish. As shown by in situ hybridization, zebrafish atp2b1a is expressed mainly in larval otic placode and lateral-line neuromast as well as in the hair cells within the adult zebrafish inner ear chamber. Zebrafish atp2b1a knockdown by antisense morpholinos reduced the number of hair cells and produced malformation of semicircular canals and smaller otoliths. These defects coincide with unbalanced body orientation. The formation of smaller otoliths in atp2b1a morphants may stem from an impairment of calcium supply in the endolymph. However, otolith formation persists in most morphants, suggesting that other zebrafish Atp2b isoforms or paracellular pathways may also transport calcium into the endolymph. These results suggest that Atp2b1a plays an important role for normal development of the auditory and vestibular system as well as calcium transport in the inner ear of zebrafish. PMID- 19218515 TI - A complex mechanism of call recognition in the katydid Neoconocephalus affinis (Orthoptera: Tettigoniidae). AB - Acoustic pattern recognition is important for bringing together males and females in many insect species. We used phonotaxis experiments on a walking compensator to study call recognition in the katydid Neoconocephalus affinis, a species with a double-pulsed call and an atypically slow pulse rate for the genus. Call recognition in this species is unusual because females require the presence of two alternating pulse amplitudes in the signal. A Fourier analysis of the stimulus-envelopes revealed that females respond only when both the first and second harmonics of the AM spectrum are of similar amplitude. The second harmonic is generated by the amplitude difference between the two pulses making up a pulse pair. Females respond to double pulses that have been merged into a single pulse only if this amplitude modulation is preserved. Further experiments suggest that females use a resonance mechanism to recognize the pulse rate of the call, supporting a neural model of rate recognition in which periodic oscillations in membrane potential are used to filter the pulse rate of the signal. Our results illustrate how a reduction in pulse rate extends the opportunities for females to evaluate fine-scale temporal properties of calls, and provide further evidence for the importance of oscillatory membrane properties in temporal processing. The results are discussed with regard to evolutionary changes in call recognition mechanisms within the genus. PMID- 19218516 TI - Continuous light affects mineralization and delays osteoid incorporation in vertebral bone of Atlantic salmon (Salmo salar L.). AB - In order to study the effects of photoperiod on fish bone, Atlantic salmon (Salmo salar L.) were exposed to two light regimes (natural and continuous light) from January until June. During the experimental period, several parameters related to the inorganic (minerals) and organic (osteoid) phases were measured. Changes in the organic phase were related to mechanical strength (yield-load) and the expression of the genes sonic hedgehog (shh) and collagen type I alpha 2 (col I). Co-variation between yield-load and the expression of both shh and col I were detected in both groups. It was also shown that fish on the continuous light regime had delayed activation of osteoid incorporation. Mineralization properties were measured with stiffness, mineral incorporation per day and expression of alkaline phosphatase (alp) and matrix Gla protein (mgp). Stiffness, mineral incorporation and gene expression followed the same trend in both light groups in late spring, whereas an increase in the expression of mgp and alp was detected in April, followed by significantly higher stiffness at last sampling in both light groups. These results indicate that constant light affects mineralization and delays osteoid incorporation in Atlantic salmon during the spring. However, in this experiment light treatment did not promote the development of vertebral deformities. Our results also suggest that shh can be used as a marker of osteoblast proliferation and col I a marker of osteoid incorporation, and that both alp and mgp expression could be associated with a rapid increase in mineralization in Atlantic salmon vertebrae. PMID- 19218517 TI - Phototropic growth in a reef flat acroporid branching coral species. AB - Many terrestrial plants form complex morphological structures and will alter these growth patterns in response to light direction. Similarly reef building corals have high morphological variation across coral families, with many species also displaying phenotypic plasticity across environmental gradients. In particular, the colony geometry in branching corals is altered by the frequency, location and direction of branch initiation and growth. This study demonstrates that for the branching species Acropora pulchra, light plays a key role in axial polyp differentiation and therefore axial corallite development--the basis for new branch formation. A. pulchra branches exhibited a directional growth response, with axial corallites only developing when light was available, and towards the incident light. Field experimentation revealed that there was a light intensity threshold of 45 micromol m(-2) s(-1), below which axial corallites would not develop and this response was blue light (408-508 nm) dependent. There was a twofold increase in axial corallite growth above this light intensity threshold and a fourfold increase in axial corallite growth under the blue light treatment. These features of coral branch growth are highly reminiscent of the initiation of phototropic branch growth in terrestrial plants, which is directed by the blue light component of sunlight. PMID- 19218518 TI - Characterization of amphioxus nebulin and its similarity to human nebulin. AB - Identification of a large molecule in muscle is important but difficult to approach by protein chemistry. In this study we isolated nebulin cDNA from the striated muscle of amphioxus, and characterized the C-terminal regions of nebulins from other chordates. Although the sequence homology with that of human is only 26%, the C-terminal region of amphioxus nebulin has similar structural motifs of 35 amino acid nebulin repeats and an SH3 domain. Using in situ indirect immunofluorescence analysis with a specific antibody raised to the bacterially produced recombinant peptide, we identified that this nebulin fragment is located in the Z-line of the sarcomere, similar to human nebulin. Pull-down and co sedimentation assays in vitro showed that the C-terminal region binds to actin, alpha-actinin and connectin (titin). These results suggest that the C-terminal region of amphioxus nebulin plays a similar role in maintaining striated muscle structure to that of human nebulin. This is the first report of the exact location of nebulin in amphioxus muscle. PMID- 19218519 TI - Skeletal muscle fiber types in the ghost crab, Ocypode quadrata: implications for running performance. AB - Ghost crabs possess rapid running capabilities, which make them good candidates for comparing invertebrate exercise physiology with that of more extensively studied vertebrates. While a number of studies have examined various aspects of running physiology and biomechanics in terrestrial crabs, none to date have defined the basic skeletal muscle fiber types that power locomotion. In the current study, we investigated skeletal muscle fiber types comprising the extensor and flexor carpopodite muscles in relation to running performance in the ghost crab. We used kinematic analyses to determine stride frequency and muscle shortening velocity and found that both parameters are similar to those of comparably sized mammals but slower than those observed in running lizards. Using several complementary methods, we found that the muscles are divided into two primary fiber types: those of the proximal and distal regions possess long sarcomeres (6.2+/-2.3 microm) observed in crustacean slow fibers and have characteristics of aerobic fibers whereas those of the muscle mid-region have short sarcomeres (3.5+/-0.4 microm) characteristic of fast fibers and appear to be glycolytic. Each fiber type is characterized by several different myofibrillar protein isoforms including multiple isoforms of myosin heavy chain (MHC), troponin I (TnI), troponin T (TnT) and a crustacean fast muscle protein, P75. Three different isoforms of MHC are differentially expressed in the muscles, with fibers of the mid-region always co-expressing two isoforms at a 1:1 ratio within single fibers. Based on our analyses, we propose that these muscles are functionally divided into a two-geared system, with the aerobic fibers used for slow sustained activities and the glycolytic mid-region fibers being reserved for explosive sprints. Finally, we identified subtle differences in myofibrillar isoform expression correlated with crab body size, which changes by several orders of magnitude during an animal's lifetime. PMID- 19218520 TI - Using omeprazole to link the components of the post-prandial alkaline tide in the spiny dogfish, Squalus acanthias. AB - After a meal, dogfish exhibit a metabolic alkalosis in the bloodstream and a marked excretion of basic equivalents across the gills to the external seawater. We used the H(+), K(+)-ATPase pump inhibitor omeprazole to determine whether these post-prandial alkaline tide events were linked to secretion of H(+) (accompanied by Cl(-)) in the stomach. Sharks were fitted with indwelling stomach tubes for pretreatment with omeprazole (five doses of 5 mg omeprazole per kilogram over 48 h) or comparable volumes of vehicle (saline containing 2% DMSO) and for sampling of gastric chyme. Fish were then fed an involuntary meal by means of the stomach tube consisting of minced flatfish muscle (2% of body mass) suspended in saline (4% of body mass total volume). Omeprazole pre-treatment delayed the post-prandial acidification of the gastric chyme, slowed the rise in Cl(-) concentration of the chyme and altered the patterns of other ions, indicating inhibition of H(+) and accompanying Cl(-) secretion. Omeprazole also greatly attenuated the rise in arterial pH and bicarbonate concentrations and reduced the net excretion of basic equivalents to the water by 56% over 48 h. Arterial blood CO(2) pressure (Pa(CO(2))) and plasma ions were not substantially altered. These results indicate that elevated gastric H(+) secretion (as HCl) in the digestive process is the major cause of the systemic metabolic alkalosis and the accompanying rise in base excretion across the gills that constitute the alkaline tide in the dogfish. PMID- 19218521 TI - Behavioral responses of big brown bats to dives by praying mantises. AB - Insectivorous echolocating bats face a formidable array of defenses employed by their airborne prey. One such insect defense is the ultrasound-triggered dive, which is a sudden, rapid drop in altitude, sometimes all the way to the ground. Although many previous studies have investigated the dynamics of such dives and their effect on insect survival rate, there has been little work on how bats may adapt to such an insect defense employed in the middle of pursuit. In this study we investigated how big brown bats (Eptesicus fuscus) adjust their pursuit strategy when flying praying mantises (Parasphendale agrionina) execute evasive, ultrasound-triggered dives. Although the mantis dive occasionally forced the bat to completely abort its chase (25% trials), in a number of cases (75% trials) the bat followed the mantis into the dive. In such cases the bat kept its sonar beam locked onto the target and maneuvered to maintain the same time efficient strategy it adopted during level flight pursuit, though it was ultimately defeated by the dive. This study suggests that although the mantis dive can be effective in evading the bat, it does not always deter the bat from continuing pursuit and, given enough altitude, the bat can potentially capture diving prey using the same flight strategy it employs to intercept prey in level flight. PMID- 19218522 TI - Urea transporter and glutamine synthetase regulation and localization in gulf toadfish gill. AB - The goal of the present study was to investigate the role of circulating cortisol and urea in the transcriptional regulation of branchial glutamine synthetase (GS), which incorporates NH(3) into glutamate to form glutamine, and the toadfish urea transporter, tUT, which is involved in urea excretion across the gill of the gulf toadfish. GS (of which there are two isoforms, LGS and GGS) and tUT mRNA expression and activity were measured in toadfish exposed to treatments that would induce variable stress responses. In addition, the role of circulating urea in tUT regulation was investigated by infusing toadfish with urea alone or in combination with intraperitoneal injection of RU486, a corticosteroid type II receptor antagonist. There was a 4.8-fold upregulation in the mRNA expression of the gill-specific GS isoform (GGS) in response to cortisol infusion and a similar upregulation in the more ubiquitous isoform (LGS). Furthermore, there was a significant 1.9-fold and 3.3-fold upregulation in the mRNA expression of the toadfish urea transporter, tUT, in response to stress through crowding or exogenous cortisol loading through infusion, respectively. In addition, tUT was found to have a urea-sensitive component to transcriptional regulation that was independent of circulating cortisol concentrations. However, the changes measured in mRNA expression of GGS, LGS and tUT did not correspond with changes in protein activity. To determine the cell type(s) involved in glutamine production and urea excretion, we attempted to localize GGS, LGS and tUT using in situ hybridization. This study is the first to show that GGS and tUT expression appear to occur in gill mitochondria-rich cells of toadfish, suggesting that these cells play a combined glutamine production and urea excretion role, which may have implications for predator avoidance. PMID- 19218523 TI - Walking, running and the evolution of short toes in humans. AB - The phalangeal portion of the forefoot is extremely short relative to body mass in humans. This derived pedal proportion is thought to have evolved in the context of committed bipedalism, but the benefits of shorter toes for walking and/or running have not been tested previously. Here, we propose a biomechanical model of toe function in bipedal locomotion that suggests that shorter pedal phalanges improve locomotor performance by decreasing digital flexor force production and mechanical work, which might ultimately reduce the metabolic cost of flexor force production during bipedal locomotion. We tested this model using kinematic, force and plantar pressure data collected from a human sample representing normal variation in toe length (N=25). The effect of toe length on peak digital flexor forces, impulses and work outputs was evaluated during barefoot walking and running using partial correlations and multiple regression analysis, controlling for the effects of body mass, whole-foot and phalangeal contact times and toe-out angle. Our results suggest that there is no significant increase in digital flexor output associated with longer toes in walking. In running, however, multiple regression analyses based on the sample suggest that increasing average relative toe length by as little as 20% doubles peak digital flexor impulses and mechanical work, probably also increasing the metabolic cost of generating these forces. The increased mechanical cost associated with long toes in running suggests that modern human forefoot proportions might have been selected for in the context of the evolution of endurance running. PMID- 19218524 TI - AMP-activated protein kinase (AMPK) in the rock crab, Cancer irroratus: an early indicator of temperature stress. AB - Exposure of marine invertebrates to high temperatures leads to a switch from aerobic to anaerobic metabolism, a drop in the cellular ATP concentration ([ATP]), and subsequent death. In mammals, AMP-activated protein kinase (AMPK) is a major regulator of cellular [ATP] and activates ATP-producing pathways, while inhibiting ATP-consuming pathways. We hypothesized that temperature stress in marine invertebrates activates AMPK to provide adequate concentrations of ATP at increased but sublethal temperatures and that AMPK consequently can serve as a stress indicator (similar to heat shock proteins, HSPs). We tested these hypotheses through two experiments with the rock crab, Cancer irroratus. First, crabs were exposed to a progressive temperature increase (6 degrees C h(-1)) from 12 to 30 degrees C. AMPK activity, total AMPK protein and HSP70 levels, reaction time, heart rate and lactate accumulation were measured in hearts at 2 degrees C increments. AMPK activity remained constant between 12 and 18 degrees C, but increased up to 9.1(+/-1.5)-fold between 18 and 30 degrees C. The crabs' reaction time also decreased above 18 degrees C. By contrast, HSP70 (total and inducible) and total AMPK protein expression levels did not vary significantly over this temperature range. Second, crabs were exposed for up to 6 h to the sublethal temperature of 26 degrees C. This prolonged exposure led to a constant elevation of AMPK activity and levels of HSP70 mRNA. AMPK mRNA continuously increased, indicating an additional response in gene expression. We conclude that AMPK is an earlier indicator of temperature stress in rock crabs than HSP70, especially during the initial response to high temperatures. We discuss the temperature dependent increase in AMPK activity in the context of Shelford's law of tolerance. Specifically, we describe AMPK activity as a cellular marker that indicates a thermal threshold, called the pejus temperature, T(p). At T(p) the animals leave their optimum range and enter a temperature range with a limited aerobic scope for exercise. This T(p) is reached periodically during annual temperature fluctuations and has higher biological significance than earlier described critical temperatures, at which the animals switch to anaerobic metabolism and HSP expression is induced. PMID- 19218525 TI - Flight variability in the woodwasp Sirex noctilio (Hymenoptera: Siricidae): an analysis of flight data using wavelets. AB - We describe flight variability in the woodwasp Sirex noctilio Fabricius, 1793 (Hymenoptera: Siricidae) by studying tethered females in a flight mill device and analyzing output data by a time series methodology. Twenty-eight wasps were flown during 24 h-long periods, under controlled temperature and lighting conditions. The maximum distance recorded was 49 km, and mean velocity was 0.37 m s(-1). All wasps lost weight during flight (mean weight loss of 10.0% of initial body mass). By using a wavelets analysis on the flight mill time series output, we identified three distinct flight patterns: regular (long acceleration-deceleration spells), periodic (alternation of acceleration-deceleration spells without resting) and pulsating (resting spells interrupted by bursts of flight activity). The first two flight patterns are indistinguishable using traditional flight mill data analysis. Flight patterns for each individual were significantly dependent on wasp body mass, suggesting a relationship with the resources used in flight and their availability. Large females flew sequentially through a regular-periodic pulsating sequence but medium sized wasps flew mostly with periodic and pulsating patterns. The smallest wasps flew only in a pulsating pattern, being incapable of long, sustained flight. Variability in size and behavior can have significant consequences on population dynamics by determining local and regional dispersal. An important outcome of our work is the introduction of wavelet analysis to study tethered flight data series for the first time. This methodology allowed us to uncover and statistically test individual variability in insect flight characteristics. PMID- 19218527 TI - Controversies in neonatal seizure management. AB - Seizures in the newborn period are common and frequently indicate serious underlying brain injury. Although accumulating evidence suggests that they may impair brain development, there are currently no evidence-based guidelines for evaluation and management of neonatal seizures. In this review, we will address some of the current controversies facing child neurologists and neonatologists, including how to define, monitor, and treat neonatal seizures. PMID- 19218526 TI - The relationships between muscle, external, internal and joint mechanical work during normal walking. AB - Muscle mechanical work is an important biomechanical quantity in human movement analyses and has been estimated using different quantities including external, internal and joint work. The goal of this study was to investigate the relationships between these traditionally used estimates of mechanical work in human walking and to assess whether they can be used as accurate estimates of musculotendon and/or muscle fiber work. A muscle-actuated forward dynamics walking simulation was generated to quantify each of the mechanical work measures. Total joint work (i.e. the time integral of absolute joint power over a full gait cycle) was found to underestimate total musculotendon work due to agonist-antagonist co-contractions, despite the effect of biarticular muscle work and passive joint work, which acted to decrease the underestimation. We did find that when the net passive joint work over the gait cycle is negligible, net joint work (i.e. the time integral of net joint power) was comparable to the net musculotendon work (and net muscle fiber work because net tendon work is zero over a complete gait cycle). Thus, during walking conditions when passive joint work is negligible, net joint work may be used as an estimate of net muscle work. Neither total external nor total internal work (nor their sum) provided a reasonable estimate of total musculotendon work. We conclude that joint work is limited in its ability to estimate musculotendon work, and that external and internal work should not be used as an estimation of musculotendon work. PMID- 19218528 TI - Multifaceted approach to determine the antagonist molecular mechanism and interaction of ibodutant ([1-(2-phenyl-1R-[[1-(tetrahydropyran-4-ylmethyl) piperidin-4-ylmethyl]-carbamoyl]-ethylcarbamoyl)-cyclopentyl]-amide) at the human tachykinin NK2 receptor. AB - Ibodutant (MEN15596, [1-(2-phenyl-1R-[[1-(tetrahydropyran-4-ylmethyl)-piperidin-4 ylmethyl]-carbamoyl]-ethylcarbamoyl)-cyclopentyl]-amide) is a tachykinin NK(2) receptor (NK(2)R) antagonist currently under phase II clinical trials for irritable bowel syndrome. This study focuses on the ibodutant pharmacodynamic profile at the human NK(2)R and compares it with two other antagonists, nepadutant (MEN11420, (cyclo-[[Asn(beta-D-GlcNAc)-Asp-Trp-Phe-Dpr-Leu]cyclo(2beta 5beta)]) and saredutant [SR48968, (S)-N-methyl-N[4-(4-acetylamino-4 phenylpiperidino)-2-(3,4-dichlorophenyl)butyl]benzamide]. In functional experiments (phosphatidylinositol accumulation) in Chinese hamster ovary cells expressing the human NK(2)R, ibodutant potency measured toward concentration response curves to neurokinin A as pK(B) was 10.6, and its antagonism mechanism was surmountable and competitive. In the same assay, antagonism equilibration and reversibility experiments of receptor blockade indicated that ibodutant quickly attains equilibrium and that reverts from receptor compartment in a slower manner. Kinetic properties of ibodutant were assessed through competitive binding kinetics experiments performed at [(3)H]nepadutant and [(3)H]saredutant binding sites. Determined K(on) and K(off) values indicated a fast association and slow dissociation rate of ibodutant at the different antagonist binding sites. Last, by radioligand binding experiments at some mutated human tachykinin NK(2)Rs, the amino acidic determinants crucial for the high affinity of ibodutant were identified at the transmembrane (TM) level: Cys167 in TM4; Ile202 and Tyr206 in TM5; Phe270, Tyr266, and Trp263 in TM6; and Tyr289 in TM7. These results indicated an extended antagonist binding pocket in the TM portion of the receptor, which is conceived crucial for TM3 and 6 arrangement and leads to G protein-coupled receptor activation. By combining this information and molecular modeling, the docking mode of ibodutant-human NK(2)R complex is proposed. PMID- 19218529 TI - Pharmacological preconditioning of mesenchymal stem cells with trimetazidine (1 [2,3,4-trimethoxybenzyl]piperazine) protects hypoxic cells against oxidative stress and enhances recovery of myocardial function in infarcted heart through Bcl-2 expression. AB - Stem cell transplantation is a possible therapeutic option to repair ischemic damage to the heart. However, it is faced with a number of challenges including the survival of the transplanted cells in the ischemic region. The present study was designed to use stem cells preconditioned with trimetazidine (1-[2,3,4 trimethoxybenzyl]piperazine; TMZ), a widely used anti-ischemic drug for treating angina in cardiac patients, to increase the rate of their survival after transplantation. Bone marrow-derived rat mesenchymal stem cells (MSCs) were subjected to a simulated host tissue environment by culturing them under hypoxia (2% O(2)) and using hydrogen peroxide (H(2)O(2)) to induce oxidative stress. MSCs were preconditioned with 10 microM TMZ for 6 h followed by treatment with 100 microM H(2)O(2) for 1 h and characterized for their cellular viability and metabolic activity. The preconditioned cells showed a significant protection against H(2)O(2)-induced loss of cellular viability, membrane damage, and oxygen metabolism accompanied by a significant increase in HIF-1alpha, survivin, phosphorylated Akt (pAkt), and Bcl-2 protein levels and Bcl-2 gene expression. The therapeutic efficacy of the TMZ-preconditioned MSCs was evaluated in an in vivo rat model of myocardial infarction induced by permanent ligation of left anterior descending coronary artery. A significant increase in the recovery of myocardial function and up-regulation of pAkt and Bcl-2 levels were observed in hearts transplanted with TMZ-preconditioned cells. This study clearly demonstrated the potential benefits of pharmacological preconditioning of MSCs with TMZ for stem cell therapy for repairing myocardial ischemic damage. PMID- 19218530 TI - Involution of collagen-induced arthritis with an angiogenesis inhibitor, PPI 2458. AB - Pannus formation, in both rheumatoid arthritis (RA) and collagen-induced arthritis (CIA), is angiogenesis-dependent. PPI-2458 [(1R)-1-carbamoyl-2-methyl] carbamic acid-(3R,3S,5S, 6R)-5-methoxy-4-[(2R,3R)-2-methyl-3-(3-methyl-but-2 enyl)oxiranyl]-1-oxaspiro(2*5)oct-6-yl ester], a new fumagillin derivative known to inhibit methionine aminopeptidase 2 (MetAP-2) and endothelial proliferation at the late G(1) phase, was evaluated in CIA rats to study its potential to involute synovitis. Arthritic syngeneic LOU rats received either a vehicle control or various dosages of oral, intravenous, or subcutaneous PPI-2458. Plasma samples were analyzed to determine a pharmacokinetic profile of PPI-2458, and whole blood was evaluated by flow cytometry to assess the effect on lymphocyte subsets. At 15 mg/kg i.v., 30 mg/kg s.c., or 100 mg/kg p.o., there was a significant reduction in clinical severity scores (p < 0.001) and blinded radiographic scores (p < 0.001) compared with vehicle control groups. Structural damage was virtually eliminated with PPI-2458. Continuous inhibition of MetAP-2 was needed to maintain benefits, although pannus involution could be achieved with the inhibitor when escape flares occurred. Pharmacokinetic analysis after a single p.o. dose showed a rapid T(max) value of 15 min followed by biphasic elimination (t(1/2), approximately 20 min and t(1/2), approximately 5 h) and an estimated oral bioavailability of approximately 15%. Flow cytometry revealed a dose-dependent decrease in white blood cells and lymphocytes manifested as decreases in circulating CD3+ T cells and natural killer cells. PPI-2458, however, did not seem to be immunosuppressive, as determined by delayed-type hypersensitivity or IgG antibody assays. These studies indicate that the MetAP-2 inhibitor PPI-2458 can regress established CIA and that angiogenic mechanisms might be important targets in the treatment of other pannus-mediated diseases such as RA. PMID- 19218531 TI - Interferon-gamma released by gluten-stimulated celiac disease-specific intestinal T cells enhances the transepithelial flux of gluten peptides. AB - Celiac sprue is a T-cell-mediated enteropathy elicited in genetically susceptible individuals by dietary gluten proteins. To initiate and propagate inflammation, proteolytically resistant gluten peptides must be translocated across the small intestinal epithelium and presented to DQ2-restricted T cells, but the effectors enabling this translocation under normal and inflammatory conditions are not well understood. We demonstrate that a fluorescently labeled antigenic 33-mer gluten peptide is translocated intact across a T84 cultured epithelial cell monolayer and that preincubation of the monolayer with media from gluten-stimulated, celiac patient-derived intestinal T cells enhances the apical-to-basolateral flux of this peptide in a dose-dependent, saturable manner. The permeability-enhancing activity of activated T-cell media is inhibited by blocking antibodies against either interferon-gamma or its receptor and is recapitulated using recombinant interferon-gamma. At saturating levels of interferon-gamma, activated T-cell media does not further increase transepithelial peptide flux, indicating the primacy of interferon-gamma as an effector of increased epithelial permeability during inflammation. Reducing the assay temperature to 4 degrees C reverses the effect of interferon-gamma but does not reduce basal peptide flux occurring in the absence of interferon-gamma, suggesting active transcellular transport of intact peptides is increased during inflammation. A panel of disease-relevant gluten peptides exhibited an inverse correlation between size and transepithelial flux but no apparent sequence constraints. Anti-interferon-gamma therapy may mitigate the vicious cycle of gluten-induced interferon-gamma secretion and interferon-gamma-mediated enhancement of gluten peptide flux but is unlikely to prevent translocation of gluten peptides in the absence of inflammatory conditions. PMID- 19218532 TI - HistoneHits: a database for histone mutations and their phenotypes. AB - Histones are the basic protein components of nucleosomes. They are among the most conserved proteins and are subject to a plethora of post-translational modifications. Specific histone residues are important in establishing chromatin structure, regulating gene expression and silencing, and responding to DNA damage. Here we present HistoneHits, a database of phenotypes for systematic collections of histone mutants. This database combines assay results (phenotypes) with information about sequences, structures, post-translational modifications, and evolutionary conservation. The web interface presents the information through dynamic tables and figures. It calculates the availability of data for specific mutants and for nucleosome surfaces. The database currently includes 42 assays on 677 mutants multiply covering 405 of the 498 residues across yeast histones H3, H4, H2A, and H2B. We also provide an interface with an extensible controlled vocabulary for research groups to submit new data. Preliminary analyses confirm that mutations at highly conserved residues and modifiable residues are more likely to generate phenotypes. Buried residues and residues on the lateral surface tend to generate more phenotypes, while tail residues generate significantly fewer phenotypes than other residues. Yeast mutants are cross referenced with known human histone variants, identifying a position where a yeast mutant causes loss of ribosomal silencing and a human variant increases breast cancer susceptibility. All data sets are freely available for download. PMID- 19218533 TI - Breakpoint graphs and ancestral genome reconstructions. AB - Recently completed whole-genome sequencing projects marked the transition from gene-based phylogenetic studies to phylogenomics analysis of entire genomes. We developed an algorithm MGRA for reconstructing ancestral genomes and used it to study the rearrangement history of seven mammalian genomes: human, chimpanzee, macaque, mouse, rat, dog, and opossum. MGRA relies on the notion of the multiple breakpoint graphs to overcome some limitations of the existing approaches to ancestral genome reconstructions. MGRA also generates the rearrangement-based characters guiding the phylogenetic tree reconstruction when the phylogeny is unknown. PMID- 19218536 TI - Screening for chronic kidney disease in emerging countries: feasibility and hurdles. PMID- 19218534 TI - Global distribution of genomic diversity underscores rich complex history of continental human populations. AB - Characterizing patterns of genetic variation within and among human populations is important for understanding human evolutionary history and for careful design of medical genetic studies. Here, we analyze patterns of variation across 443,434 single nucleotide polymorphisms (SNPs) genotyped in 3845 individuals from four continental regions. This unique resource allows us to illuminate patterns of diversity in previously under-studied populations at the genome-wide scale including Latin America, South Asia, and Southern Europe. Key insights afforded by our analysis include quantifying the degree of admixture in a large collection of individuals from Guadalajara, Mexico; identifying language and geography as key determinants of population structure within India; and elucidating a north south gradient in haplotype diversity within Europe. We also present a novel method for identifying long-range tracts of homozygosity indicative of recent common ancestry. Application of our approach suggests great variation within and among populations in the extent of homozygosity, suggesting both demographic history (such as population bottlenecks) and recent ancestry events (such as consanguinity) play an important role in patterning variation in large modern human populations. PMID- 19218537 TI - The vaptans ante portas: a status report. PMID- 19218539 TI - The value of QuantiFERON TB-Gold in the diagnosis of tuberculosis among dialysis patients. AB - BACKGROUND: It is difficult to diagnose tuberculosis (TB) in dialysis patients because of the high rate of extrapulmonary TB in these patients compared with the general population. Recently, a new diagnostic test called QuantiFERON (QFT) has been developed and shown promise as a diagnostic tool for active TB diseases and latent TB infection. METHODS: We examined 162 dialysis patients admitted to a single institute, including 8 patients with active TB, and evaluated the utility of this test in dialysis patients. RESULTS: Among 162 dialysis patients, positive QFT results occurred in 28 (17.3%), negative QFT results occurred in 95 (58.6%) and indeterminate QFT results occurred in 39 (24.1%). All eight active TB patients had positive QFT results, and none of the 95 patients with negative results had active TB. Among 23 patients with a history of active TB, 10 (43.5%) had positive results. Although the indeterminate rate was relatively high, no patient with an indeterminate result had active TB. Factors such as shorter duration of dialysis, lower lymphocyte count and higher white blood cell count were associated with indeterminate results. Among 105 cases after excluding the patients with previous TB or indeterminate results, the sensitivity of the QFT is 100% (8 of 8) and the specificity is 89.7% (87 of 97 cases). CONCLUSIONS: Our data suggest that the QFT test is a useful supplementary tool for the diagnosis of active TB even in dialysis patients. Negative and indeterminate results on this test may be used to exclude the presence of active TB. PMID- 19218538 TI - Fabry disease: progression of nephropathy, and prevalence of cardiac and cerebrovascular events before enzyme replacement therapy. AB - BACKGROUND: In Fabry disease, progressive glycolipid accumulation leads to organ damage and early demise, but the incidence of renal, cardiac and cerebrovascular events has not been well characterized. METHODS: We conducted a retrospective chart review of 279 affected males and 168 females from 27 sites (USA, Canada, Europe). The pre-defined study endpoints included progression of renal, cardiac and cerebrovascular involvement and/or death before the initiation of enzyme replacement therapy. RESULTS: The mean rate of estimated glomerular filtration rate (eGFR) decline for patients was -2.93 for males, and -1.02 ml/min/1.73 m(2)/year for females. Prevalence and severity of proteinuria, baseline eGFR <60 ml/min/1.73 m(2) and hypertension were associated with more rapid loss of eGFR. Advanced Fabry nephropathy was more prevalent and occurred earlier among males than females. Cardiac events (mainly arrhythmias), strokes and transient ischaemic attacks occurred in 49, 11, 6% of males, and in 35, 8, 4% of females, respectively. The mean age at death for 20 male patients was 49.9 years. CONCLUSIONS: Baseline proteinuria, reduced baseline eGFR, hypertension and male gender were associated with more rapid progression of Fabry nephropathy. The eGFR progression rate may increase with advancing nephropathy, and may differ between subgroups of patients with Fabry disease. PMID- 19218540 TI - The Hannover Dialysis Outcome study: comparison of standard versus intensified extended dialysis for treatment of patients with acute kidney injury in the intensive care unit. AB - BACKGROUND: Increasing the dose of renal replacement therapy has been shown to improve survival in critically ill patients with acute kidney injury (AKI) in several smaller European trials. However, a very recent large multicentre trial in the USA could not detect an effect of dose of renal replacement therapy on mortality. Based on those studies, it is not known whether a further increase in dialysis dose above and beyond the currently employed doses would improve survival in patients with AKI. We therefore aimed to assess mortality and renal recovery of patients with AKI receiving either standard (SED) or intensified extended dialysis (IED) therapy in the intensive care unit. METHODS: A prospective randomized parallel group study was conducted in seven intensive care units of a tertiary university hospital. Pre-existing chronic kidney disease was an exclusion criterion. A total of 156 patients (570 screened) with AKI requiring renal replacement therapy were randomly assigned to receive standard dialysis [dosed to maintain plasma urea levels between 120 and 150 mg/dL (20-25 mmol/L)] or intensified dialysis [dosed to maintain plasma urea levels <90 mg/dL (<15 mmol/L)]. Outcome measures were survival at Day 14 (primary) and survival and renal recovery at Day 28 (secondary) after initiation of renal replacement therapy. RESULTS: Treatment intensity differed significantly (P < 0.01 for plasma urea and administered dose). No differences between intensified and standard treatment were seen for survival by Day 14 (70.4% versus 70.7%) or Day 28 (55.6% versus 61.3%), or for renal recovery amongst the survivors by Day 28 (60.0% versus 63.0%). CONCLUSIONS: Although this study cannot deliver a definitive answer, it suggests that increasing the dose of extended dialysis above the currently recommended dose might neither reduce mortality nor improve renal recovery in critically ill patients, mainly septic patients, with AKI. PMID- 19218541 TI - The soft underbelly of research in the psychosocial treatment of ADHD. PMID- 19218542 TI - Long-term effectiveness and safety of dexmethylphenidate extended-release capsules in adult ADHD. AB - OBJECTIVE: This study evaluates dexmethylphenidate extended release (d-MPH-ER) in adults with ADHD. METHOD: Following a 5-week, randomized, controlled, fixed-dose study of d-MPH-ER 20 to 40 mg/d, 170 adults entered a 6-month open-label extension (OLE) to assess long-term safety, with flexible dosing of 20 to 40 mg/d. Exploratory effectiveness outcomes included change from Week 5 on ADHD Rating Scale (ADHD-RS) and proportion of responders on Clinical Global Impressions-Improvement (CGI-I) scale. RESULTS: 103 patients completed OLE, and effectiveness was evaluable in 102 patients. d-MPH-ER was well tolerated; the most common adverse events (>15%) were headache, insomnia, and decreased appetite. Mean improvements in ADHD-RS score were -10.2 for patients switched from placebo to d-MPH-ER (n = 20) and -8.4 for those maintained on d-MPH-ER (n = 82). Respective CGI-I responder rates were 95.0% and 95.1%. CONCLUSION: Once daily d-MPH-ER 20 to 40 mg is safe and effective for long-term treatment of adult ADHD. PMID- 19218543 TI - Academic performance in ADHD when controlled for comorbid learning disorders, family income, and parental education in Brazil. AB - OBJECTIVE: Scholastic achievement in a nonclinical sample of ADHD children and adolescents was evaluated taking into consideration variables such as comorbid learning disorders, family income, and parental education which may also be associated with poor academic performance. METHOD: After screening for ADHD in 396 students, the authors compared academic performance of 26 ADHD individuals and 31 controls paired for gender, age, and intelligence level considering both mathematics and Portuguese language scores. Learning disorders were investigated and the Diagnostic and Statistical Manual of Mental Disorders (4th ed.; DSM-IV ) criteria were met using structured interviews. RESULTS: The prevalence of academic underachievement was 2.98 times higher in students with ADHD, the most frequent subtype being predominantly inattentive. Parental educational level, family income, and comorbid learning disorders could not explain the discrepancies between ADHD students and controls. CONCLUSIONS: ADHD seems to be associated with poor academic performance even in the absence of comorbid learning disorders, lower family income, and parental educational level. PMID- 19218544 TI - Comparative study of children with ADHD only, autism spectrum disorder + ADHD, and chronic multiple tic disorder + ADHD. AB - OBJECTIVE: Identification of differences among children with ADHD only, autism spectrum disorder (ASD)+ADHD, and chronic multiple tic disorder (CMTD)+ADHD may lead to better understanding of clinical phenotypes. METHOD: Children were evaluated using the parent- and teacher-completed questionnaires. RESULTS: All three groups were highly similar in severity of oppositional defiant disorder and conduct disorder symptoms; however, the ASD+ADHD group generally exhibited the most severe anxiety, although the CMTD+ADHD group had the most severe generalized anxiety. The two comorbid groups had the most involved medical histories and the greatest likelihood of a family history of psychopathology. CONCLUSION: Groups differed in clinically meaningful ways, and the apparent association between tics and anxiety may explain in part the elevated levels of anxiety in both comorbid groups. Collectively, results suggest that ADHD may be better conceptualized as a family of interrelated syndromes defined in part by comorbid conditions and that continued research is clearly warranted. PMID- 19218545 TI - A descriptive analysis of transitional housing programs for survivors of intimate partner violence in the United States. AB - The Violence Against Women Act (VAWA) of 2005 focuses on safe and independent housing for survivors of intimate partner violence (IPV). The focus on housing in the latest version of VAWA suggests recognition by Congress that removing barriers and increasing access to safe housing is critical to our nation's response to IPV, and that this type of systems-level response is necessary to reduce the link between IPV and subsequent homelessness. This study examines the current state of transitional housing programs (THPs) and discusses future program considerations, including the need for evaluation studies that consider the possible impact that transitional housing programs have on the rates of violence toward women and their children, and on women's ability to achieve economic stability after separating from their abusive partners. PMID- 19218547 TI - Role of ephrinB2 in nonproductive angiogenesis induced by Delta-like 4 blockade. AB - Delta-like 4 (DLL4) is one of the Notch ligands and plays an important role in vascular development. DLL4 blockade inhibits tumor growth by promoting nonproductive angiogenesis, which is characterized by an increase in vascular density and decrease in tissue perfusion. However, a detailed mechanism remains unclear. In this study, newly developed neutralizing antibodies against mouse and human DLL4 were used to investigate the possible involvement of VEGF-DLL4 ephrinB2 cascade in nonproductive angiogenesis caused by DLL4 blockade. DLL4 blockade and soluble ephrinB2 treatment suppressed tumor growth and induced nonproductive angiogenesis. DLL4 was expressed in subcutaneous tumors, and DLL4 blockade suppressed ephrinB2 expression in the tumors. DLL4 blockade significantly promoted human umbilical vein endothelial cell (HUVEC) proliferation in vitro, and the effect was additive to that of VEGF. Both DLL4 blockade and VEGF significantly increased cord length and branch points in a tubular formation assay. Expression of ephrinB2 in HUVECs was enhanced by VEGF alone, and the enhancement was inhibited by DLL4 blockade. Moreover, when we studied the effect of ephrinB2 RNA interference on HUVEC tubular formation, knockdown of ephrinB2 mimicked the effect of DLL4. These results suggest that ephrinB2 plays a crucial role in nonproductive angiogenesis caused by DLL4 blockade. PMID- 19218548 TI - Aberrant expression of the homeobox gene CDX2 in pediatric acute lymphoblastic leukemia. AB - Members of the caudal (cdx) family of homeobox proteins are essential regulators of embryonic blood development in zebrafish. Previously, we reported that the murine homologues (Cdx1, Cdx2, and Cdx4) affect formation and differentiation of embryonic stem cell (ESC)-derived hematopoietic progenitor cells. Consistent with the notion that embryonic pathways can reactivate during adult oncogenesis, recent studies suggest involvement of CDX2 in human acute myeloid leukemia (AML). Here we study CDX2 in healthy and leukemic human lymphoid cells, and show that a majority of leukemic samples display various degrees of aberrant CDX2 expression. Analysis of a cohort of 37 childhood acute lymphoblastic leukemia (ALL) patients treated in our hospital reveals that high CDX2 expression levels at diagnosis correlate with persistence of minimal residual disease (MRD) during the course of treatment. Thus, CDX2 expression levels may serve as a marker for adverse prognosis in pediatric ALL. PMID- 19218546 TI - C-terminal ADAMTS-18 fragment induces oxidative platelet fragmentation, dissolves platelet aggregates, and protects against carotid artery occlusion and cerebral stroke. AB - Anti-platelet integrin GPIIIa49-66 antibody (Ab) induces complement-independent platelet oxidative fragmentation and death by generation of platelet peroxide following NADPH oxidase activation. A C-terminal 385-amino acid fragment of ADAMTS-18 (a disintegrin metalloproteinase with thrombospondin motifs produced in endothelial cells) induces oxidative platelet fragmentation in an identical kinetic fashion as anti-GPIIIa49-66 Ab. Endothelial cell ADAMTS-18 secretion is enhanced by thrombin and activated by thrombin cleavage to fragment platelets. Platelet aggregates produced ex vivo with ADP or collagen and fibrinogen are destroyed by the C-terminal ADAMTS-18 fragment. Anti-ADAMTS-18 Ab shortens the tail vein bleeding time. The C-terminal fragment protects against FeCI3-induced carotid artery thrombosis as well as cerebral infarction in a postischemic stroke model. Thus, a new mechanism is proposed for platelet thrombus clearance, via platelet oxidative fragmentation induced by thrombin cleavage of ADAMTS-18. PMID- 19218549 TI - Interactions of platelet integrin alphaIIb and beta3 transmembrane domains in mammalian cell membranes and their role in integrin activation. AB - Clustering and occupancy of platelet integrin alpha(IIb)beta(3) (GPIIb-IIIa) generate biologically important signals: conversely, intracellular signals increase the integrins' affinity, leading to integrin activation; both forms of integrin signaling play important roles in hemostasis and thrombosis. Indirect evidence implicates interactions between integrin alpha and beta transmembrane domains (TMDs) and cytoplasmic domains in integrin signaling; however, efforts to directly identify these associations have met with varying and controversial results. In this study, we develop mini-integrin affinity capture and use it in combination with nuclear magnetic resonance spectroscopy to show preferential heterodimeric association of integrin alpha(IIb)beta(3) TMD tails via specific TMD interactions in mammalian cell membranes in lipid bicelles. Furthermore, charge reversal mutations at alpha(IIb)(R995)beta(3)(D723) confirm a proposed salt bridge and show that it stabilizes the TMD-tail association; talin binding to the beta(3) tail, which activates the integrin, disrupts this association. These studies establish the preferential heterodimeric interactions of integrin alpha(IIb)beta(3) TMD tails in mammalian cell membranes and document their role in integrin signaling. PMID- 19218550 TI - Genome-wide evolutionary analysis of the noncoding RNA genes and noncoding DNA of Paramecium tetraurelia. AB - The compact genome of the unicellular eukaryote Paramecium tetraurelia contains noncoding DNA (ncDNA) distributed into >39,000 intergenic sequences and >90,000 introns of 390 base pairs (bp) and 25 bp on average, respectively. Here we analyzed the molecular features of the ncRNA genes, introns, and intergenic sequences of this genome. We mainly used computational programs and comparative genomics possible because the P. tetraurelia genome had formed throughout whole genome duplications (WGDs). We characterized 417 5S rRNA, snRNA, snoRNA, SRP RNA, and tRNA putative genes, 415 of which map within intergenic sequences, and two, within introns. The evolution of these ncRNA genes appears to have mainly involved purifying selection and gene deletion. We then compared the introns that interrupt the protein-coding gene duplicates arisen from the recent WGD and identified a population of a few thousands of introns having evolved under most stringent constraints (>95% of identity). We also showed that low nucleotide substitution levels characterize the 50 and 80-115 base pairs flanking, respectively, the stop and start codons of the protein-coding genes. Lower substitution levels mark the base pairs flanking the highly transcribed genes, or the start codons of the genes of the sets with a high number of WGD-related sequences. Finally, adjacent to protein-coding genes, we characterized 32 DNA motifs able to encode stable and evolutionary conserved RNA secondary structures and defining putative expression controlling elements. Fourteen DNA motifs with similar properties map distant from protein-coding genes and may encode regulatory ncRNAs. PMID- 19218551 TI - Characterization of a mimivirus RNA cap guanine-N2 methyltransferase. AB - A 2,2,7-trimethylguanosine (TMG) cap is a signature feature of eukaryal snRNAs, telomerase RNAs, and trans-spliced nematode mRNAs. TMG and 2,7-dimethylguanosine (DMG) caps are also present on mRNAs of two species of alphaviruses (positive strand RNA viruses of the Togaviridae family). It is presently not known how viral mRNAs might acquire a hypermethylated cap. Mimivirus, a giant DNA virus that infects amoeba, encodes many putative enzymes and proteins implicated in RNA transactions, including the synthesis and capping of viral mRNAs and the promotion of cap-dependent translation. Here we report the identification, purification, and characterization of a mimivirus cap-specific guanine-N2 methyltransferase (MimiTgs), a monomeric enzyme that catalyzes a single round of methyl transfer from AdoMet to an m(7)G cap substrate to form a DMG cap product. MimiTgs, is apparently unable to convert a DMG cap to a TMG cap, and is thereby distinguished from the structurally homologous yeast and human Tgs1 enzymes. Nonetheless, we show genetically that MimiTgs is a true ortholog of Saccharomyces cerevisiae Tgs1. Our results hint that DMG caps can satisfy many of the functions of TMG caps in vivo. We speculate that DMG capping of mimivirus mRNAs might favor viral protein synthesis in the infected host. PMID- 19218553 TI - A PCR-based platform for microRNA expression profiling studies. AB - MicroRNAs (miRNAs) are newly identified small noncoding RNA molecules that play important regulatory roles in many biological processes. Changes in miRNA expression are associated with a variety of normal physiological processes as well as with human diseases including cancer. Thus, one important way to characterize miRNA function is to identify changes in miRNA expression profiles. To date, microarrays are the most widely used technique for miRNA expression profiling analysis. However, there are major limitations when microarrays are applied to miRNA profiling studies. Here we report a new PCR-based assay platform for miRNA expression profiling analysis. The primers associated with these miRNA assays were designed using a novel bioinformatics algorithm that has incorporated many primer selection features for improved assay specificity, sensitivity, and homogeneity. The primer design algorithm has been extensively validated experimentally and has been shown to have robust performance for the measurement of miRNA expression in a variety of biological samples at a low cost. PMID- 19218552 TI - Multiple roles for polypyrimidine tract binding (PTB) proteins in trypanosome RNA metabolism. AB - Trypanosomatid genomes encode for numerous proteins containing an RNA recognition motif (RRM), but the function of most of these proteins in mRNA metabolism is currently unknown. Here, we report the function of two such proteins that we have named PTB1 and PTB2, which resemble the mammalian polypyrimidine tract binding proteins (PTB). RNAi silencing of these factors indicates that both are essential for life. PTB1 and PTB2 reside mostly in the nucleus, but are found in the cytoplasm, as well. Microarray analysis performed on PTB1 and PTB2 RNAi silenced cells indicates that each of these factors differentially affects the transcriptome, thus regulating a different subset of mRNAs. PTB1 and PTB2 substrates were categorized bioinformatically, based on the presence of PTB binding sites in their 5' and 3' flanking sequences. Both proteins were shown to regulate mRNA stability. Interestingly, PTB proteins are essential for trans splicing of genes containing C-rich polypyrimidine tracts. PTB1, but not PTB2, also affects cis-splicing. The specificity of binding of PTB1 was established in vivo and in vitro using a model substrate. This study demonstrates for the first time that trans-splicing of only certain substrates requires specific factors such as PTB proteins for their splicing. The trypanosome PTB proteins, like their mammalian homologs, represent multivalent RNA binding proteins that regulate mRNAs from their synthesis to degradation. PMID- 19218554 TI - Confirming the factor structure of attention-deficit/hyperactivity disorder symptoms in college students using student and parent data. AB - This study used confirmatory factor analysis to compare one-, two-, and three factor models of attention-deficit/hyperactivity disorder (ADHD) symptoms to determine which model is the best fit for the data. Participants were 190 clinic referred college students who had been evaluated for ADHD, 155 of whom had received a diagnosis. Data consisted of both self- and other (e.g., parent) ratings of both current and childhood symptoms. Symptoms came directly from the DSM-IV criteria for ADHD. A three-factor model, consistent with the DSM-III, was superior for current and childhood symptoms, regardless of rater (i.e., self or parent). The primary implication for these findings is that there may be a viable Impulsive subtype of ADHD within the adult population. Further research might include a closer examination of the unique functional limitations associated with impulsivity, as well as the development of diagnostic items that maximize model fit. PMID- 19218556 TI - Open wedge high tibial osteotomy: a roentgenographic comparison of a horizontal and an oblique osteotomy on patellar height and sagittal tibial slope. AB - BACKGROUND: In patients with unicompartmental medial knee arthritis, medial opening wedge high tibial osteotomy is used to shift the mechanical weightbearing line laterally to reduce pain and improve function. There have been concerns that opening wedge high tibial osteotomy is associated with a reduction of patellar height and increase in the sagittal posterior tibial slope, both of which can adversely affect the final result. HYPOTHESIS: A more distal oblique osteotomy at the level of insertion of the patellar tendon should decrease these effects when compared with a horizontal osteotomy made proximal to the patellar tendon insertion. STUDY DESIGN: Cohort study; Level of evidence, 3. METHODS: Review of 22 horizontal and 19 oblique high tibial osteotomies with a mean follow-up of 4.2 +/- 1.8 years (mean +/- SD) was performed. Anatomic tibiofemoral angle, mechanical weightbearing line, medial coronal tibial plateau angle, patellar height (Blackburne and Peel ratio), and sagittal tibial slope were measured. RESULTS: In both groups, the weightbearing line was equally shifted toward the center of the plateau. In the horizontal group, the Blackburne and Peel ratio decreased from 0.85 +/- 0.16 to 0.67 +/- 0.12, and the sagittal tibial slope was increased from 7.7 degrees +/- 4.6 degrees to 10.7 degrees +/- 3.8 degrees (P < .001). In comparison, the oblique group did not show any significant postoperative changes for these 2 parameters. In the oblique group, 2 patients sustained loss of correction and early failure when the osteotomy remained below the metaphyseal flare on the lateral cortex. CONCLUSION: The oblique osteotomy group showed more normalized postoperative sagittal tibial slope and patellar height. Caution should be exercised not to osteotomize too distally. PMID- 19218555 TI - A dynamic, cytoplasmic triacylglycerol pool in enterocytes revealed by ex vivo and in vivo coherent anti-Stokes Raman scattering imaging. AB - The absorptive cells of the small intestine, enterocytes, are not generally thought of as a cell type that stores triacylglycerols (TGs) in cytoplasmic lipid droplets (LDs). We revisit TG metabolism in enterocytes by ex vivo and in vivo coherent anti-Stokes Raman scattering (CARS) imaging of small intestine of mice during dietary fat absorption (DFA). We directly visualized the presence of LDs in enterocytes. We determined lipid amount and quantified LD number and size as a function of intestinal location and time post-lipid challenge via gavage feeding. The LDs were confirmed to be primarily TG by biochemical analysis. Combined CARS and fluorescence imaging indicated that the large LDs were located in the cytoplasm, associated with the tail-interacting protein of 47 kDa. Furthermore, in vivo CARS imaging showed real-time variation in the amount of TG stored in LDs through the process of DFA. Our results highlight a dynamic, cytoplasmic TG pool in enterocytes that may play previously unexpected roles in processes, such as regulating postprandial blood TG concentrations. PMID- 19218557 TI - Tibial fixation in anterior cruciate ligament reconstruction with bone-patellar tendon-bone and semitendinosus-gracilis autografts: a comparison between bioabsorbable screws and bioabsorbable cross-pin fixation. AB - BACKGROUND: Knee instability after anterior cruciate ligament reconstruction rarely manifests with activities of daily living, but it may occur in high-level sports performance, resulting in secondary injuries. Faced with these circumstances, sports orthopaedists have continued to improve on the results obtained with surgical techniques for treating knee joint injuries. HYPOTHESIS: Transverse tibial fixation with bioabsorbable cross pins is a valid technique for anterior cruciate ligament reconstruction with both the patellar tendon (bone patellar tendon-bone) and semitendinosus and gracilis tendon. STUDY DESIGN: Cohort study; Level of evidence, 3. METHODS: The study population was 120 patients operated on by the same surgeon and subdivided into 4 subgroups of 30 subjects each. Patients were chosen and assigned to each group consecutively. Patients in group 1 (bone-patellar tendon-bone) were implanted with bioabsorbable interference screws, group 2 (bone-patellar tendon-bone) received 2 bioabsorbable cross pins, group 3 (semitendinosus and gracilis tendon) received a bioabsorbable interference screw, and group 4 (semitendinosus and gracilis tendon) received 2 bioabsorbable cross pins. Patients were assessed at 5-year follow-up for International Knee Documentation Committee Knee Ligament Standard Evaluation Form, Lysholm Knee Scoring Scale, Lysholm-Tegner Activity Scale, and KT-1000 arthrometer testing. RESULTS: The results at 5 years after anterior cruciate ligament reconstruction were similar across all 4 groups. No significant differences have been observed between groups. CONCLUSION: The results suggest that transverse tibial fixation with bioabsorbable cross pins is a safe and reliable procedure, yielding clinical results that are comparable with those of the more widely used bioabsorbable interference screws. PMID- 19218558 TI - Magnetic resonance imaging in acute traumatic and chronic meniscal tears of the knee: a diagnostic accuracy study in young adults. AB - BACKGROUND: No previous research has investigated the diagnostic validity of magnetic resonance imaging for acute versus chronic meniscal tears using comparable materials and methods. HYPOTHESIS: There is no difference in the diagnostic validity of magnetic resonance imaging for acute versus chronic meniscal tears in young adults. STUDY DESIGN: Cohort study (diagnosis); Level of evidence, 2. METHODS: A total of 628 young adult military personnel underwent magnetic resonance imaging and arthroscopy of the knee over a 6-year period. Inclusion criteria were met by 82 patients with acute knee trauma (magnetic resonance imaging within 30 days from trauma) and 40 patients with chronic knee symptoms (symptoms lasting over 6 months before magnetic resonance imaging). The original magnetic resonance imaging and arthroscopy records were reviewed twice by a musculoskeletally trained radiologist, blinded to previous magnetic resonance imaging and arthroscopy findings. Interobserver correlations and intraobserver reliability were calculated and reported. Arthroscopy served as the gold standard when calculating the diagnostic values of magnetic resonance imaging for acute and chronic meniscal tears. RESULTS: The median age of the patients was 20 years (range, 18-25). Magnetic resonance imaging detected acute meniscal tears with sensitivity of 67%, specificity of 93%, and diagnostic accuracy of 88% and chronic meniscal tears with 64%, 91%, and 86%, respectively. There was no statistically significant difference in magnetic resonance imaging results between the 2 groups. CONCLUSION: The diagnostic validity of magnetic resonance imaging is similar for meniscal tears in acute knee trauma and in knee symptoms lasting over 6 months in young adults. The results also suggest that effusion or hemarthrosis do not weaken the diagnostic validity of magnetic resonance imaging. The magnetic resonance imaging sensitivity achieved in the present study was relatively poor, but the specificity was good for both acute and chronic meniscal tears. Despite negative magnetic resonance imaging findings at the acute stage of knee trauma, patient monitoring and readiness for arthroscopy should be considered if justified by the patient's symptoms. PMID- 19218559 TI - Proximal hamstring tendinopathy: results of surgical management and histopathologic findings. AB - BACKGROUND: Tendon disorders are common problems in sports and are known to be difficult to treat. Only limited information is available concerning treatment of proximal hamstring tendinopathy. To the authors' knowledge, no histopathologic findings of proximal hamstring tendinosis have been published. HYPOTHESIS: Surgery (semimembranosus tenotomy and exploration of the sciatic nerve) is an effective treatment for proximal hamstring tendinopathy. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: A total of 103 cases of proximal hamstring tendinopathy in athletes (58 men, 32 women; 13 bilateral operations) with surgical treatment were included. The cases were retrospectively analyzed, and a 4-category rating system was used to evaluate the overall result. At the follow up, the patients were asked about possible symptoms and their return to sports. Biopsy samples from 15 of the operated tendons were taken and analyzed by a pathologist. RESULTS: The average follow-up was 49 months (range, 12-156 months). The result was evaluated to be excellent in 62 cases, good in 30, fair in 5, and poor in 6. After surgery, 80 of the 90 patients were able to return to the same level of sporting activity as before the onset of the symptoms. This took a mean of 5 months (range, 2-12 months). Typical morphologic findings of tendinosis were found in all biopsy specimens. CONCLUSION: Given the good functional outcome and low complication rate, the authors present surgical treatment as a valuable option in proximal hamstring tendinopathy if conservative treatment fails. PMID- 19218560 TI - Long-term follow-up of acute arthroscopic Bankart repair for initial anterior shoulder dislocations in young athletes. AB - BACKGROUND: Little is known of the long-term results of acute arthroscopic Bankart repair for first-time traumatic anterior glenohumeral dislocations. HYPOTHESIS: Acute arthroscopic Bankart repair for first-time traumatic anterior glenohumeral dislocations will provide good results at long-term follow-up. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: The authors evaluated a cohort of young patients who sustained first-time anterior glenohumeral dislocations and were acutely treated with arthroscopic Bankart repair using bioabsorbable tacks. Subjective outcome measures were obtained at a mean follow up of 11.7 years (range, 9.1-13.9 years). RESULTS: Thirty-nine patients (40 shoulders) were available of the original cohort of 49 shoulders (82%). Two of the 9 who were lost to follow-up had revision surgery before being lost and are carried forward in the calculations of recurrent instability and revision surgery but are not included in the calculation of the functional scores. The mean Single Assessment Numeric Evaluation was 91.7, the mean Western Ontario Shoulder Instability score was 371.7, the mean subjective Rowe score was 25.3, the mean Simple Shoulder Test was 11.1, the mean American Shoulder and Elbow Society score was 90.9, the mean Short Form-36 Physical Component score was 94.4, and the mean Tegner score was 6.5. Six patients sustained recurrent dislocations for a redislocation rate of 14.3%. Nine patients (21.4%) reported experiencing subluxation events. Six patients (14.3%) underwent revision stabilization surgery. CONCLUSION: At long-term follow-up, acute arthroscopic Bankart repair for first-time traumatic anterior glenohumeral dislocations resulted in excellent subjective function and return to athletics in young, active patients with an acceptable rate of recurrence and reoperation. PMID- 19218561 TI - Expanding treatment options for type 2 diabetes: the old and the new. AB - PURPOSE: Type 2 diabetes is characterized by multiple metabolic abnormalities including the dysfunction of pancreatic islet cells. Current treatment options have different mechanisms of action and are variably effective at lowering blood glucose levels. The newest pharmacologic treatments, incretin mimetics and dipeptidyl peptidase-IV (DPP-4) inhibitors, target the incretin system, and it is important to understand their role in current treatment paradigms and in addressing the basic pathophysiology of type 2 diabetes-pancreatic islet cell dysfunction. CONCLUSION: This article discusses the pathophysiology of type 2 diabetes including impairment of the incretin effect, and explains how incretin mimetics and DPP-4 inhibitors are being incorporated into treatment algorithms to address pancreatic islet cell dysfunction and enable patients to achieve glycemic targets. PMID- 19218562 TI - The therapeutic role of incretin mimetics and DPP-4 inhibitors. AB - PURPOSE: Gastric inhibitory peptide (GIP) and glucagon-like peptide-1 (GLP-1) stimulate the secretion of insulin when blood glucose levels are elevated and inhibit the postprandial release of glucagon. An increased understanding of the role of these incretin hormones in insulin and blood glucose regulation has resulted in the development of new types of medications for managing diabetes. GLP-1 agonists and dipeptidyl peptidase-IV (DPP-4) inhibitors are 2 classes of medications that have been shown to offer benefits for patients with type 2 diabetes. To use and teach patients how to use these medications effectively, educators need to understand their mechanisms of action, indications and contraindications, side effects, and effectiveness. Expanding the number of options available has increased the complexity of providing care and education, but also offers new opportunities to help our patients achieve better outcomes. CONCLUSION: This article provides an overview of the physiology of insulin regulation and the roles of GIP, GLP-1, and DPP-4; discusses how the action of incretin hormones is affected by diabetes; and describes the therapeutic agents in these classes and their potential use by people with diabetes. PMID- 19218563 TI - Translating clinical trial data into new patterns of practice. AB - PURPOSE: The ongoing research into the intricacies of glucoregulation and the upsurge in the number of pharmacologic therapies has broadened the treatment options for type 2 diabetes. Successful translation of pharmacologic clinical trial data into new patterns of practice begins by creating the space for a new therapy through a thorough understanding of its mechanism of action. CONCLUSION: This article will discuss important considerations for identifying patients who could benefit from incretin mimetic therapy, the role of the diabetes educator in building a cohesive team, and teaching strategies that support patients' self management efforts. PMID- 19218564 TI - Sumoylation of the transcription factor NFATc1 leads to its subnuclear relocalization and interleukin-2 repression by histone deacetylase. AB - The family of NFAT (nuclear factor of activated T-cells) transcription factors plays an important role in cytokine gene regulation. In peripheral T-cells NFATc1 and -c2 are predominantly expressed. Because of different promoter and poly(A) site usage as well as alternative splicing events, NFATc1 is synthesized in multiple isoforms. The highly inducible NFATc1/A contains a relatively short C terminus, whereas the longer, constitutively expressed isoform NFATc1/C spans an extra C-terminal peptide of 246 amino acids. Interestingly, this NFATc1/C specific terminus can be highly sumoylated. Upon sumoylation, NFATc1/C, but not the unsumoylated NFATc1/A, translocates to promyelocytic leukemia nuclear bodies. This leads to interaction with histone deacetylases followed by deacetylation of histones, which in turn induces transcriptionally inactive chromatin. As a consequence, expression of the NFATc1 target gene interleukin-2 is suppressed. These findings demonstrate that the modification by SUMO (small ubiquitin-like modifier) converts NFATc1 from an activator to a site-specific transcriptional repressor, revealing a novel regulatory mechanism for NFATc1 function. PMID- 19218567 TI - Validity of ejection fraction as a measure of myocardial functional state: impact of asynchrony. AB - AIMS: The goal of this study was to test whether peculiarities of left ventricular (LV) regional function place limits on the validity of ejection fraction (EF) as a measure of the myocardial functional state. METHODS AND RESULTS: Transthoracic and transoesophageal data from patients with a variety of cardiac conditions were used for analysis of LV regional function. The focus was on the effects of mechanical asynchrony. Ejection fraction was calculated on the basis of LV end-diastolic volume and end-systolic volume obtained by two different ways: (i) end-systolic volume as a whole; and (ii) the sum of all regional end-systolic volumes (which may occur at different times). The relative difference, D-EF, between EFs obtained by (i) and (ii) was taken as the 'merit' of EF. A value of zero is the highest merit. Irrespective of the examination method, we found that D-EF was always higher than zero, and that its value depended on the extent of mechanical asynchrony. CONCLUSIONS: Ejection fraction is not the arithmetic average of regional EFs. An increase of asynchrony increases D-EF, i.e. it reduces the merit of EF as a measure of cardiac function. PMID- 19218565 TI - Importin 13 mediates nuclear import of histone fold-containing chromatin accessibility complex heterodimers. AB - The histone fold is a structural element that facilitates heterodimerization, and histone fold heterodimers play crucial roles in gene regulation. Here, we investigated the nuclear import of two human histone fold pairs, which belong to the H2A/H2B family: CHRAC-15/CHRAC-17 and p12/CHRAC-17. Our results from in vitro nuclear import assays with permeabilized cells and in vivo cotransfection experiments reveal that importin 13 facilitates nuclear import of both histone fold heterodimers. Using glutathione S-transferase pulldown experiments, we provide evidence that heterodimers are required for efficient binding of importin 13 because the monomers alone do not significantly interact. Mutational analysis shows that stepwise substitution of basic amino acid residues conserved among the histone fold subunits leads to a progressive loss of importin 13 binding and nuclear accumulation of CHRAC-15/CHRAC-17 and p12/CHRAC-17. The distribution of basic amino acid residues among the histone fold subunits essential for nuclear uptake suggests that heterodimerization of the histone fold motif-containing proteins forms an importin 13-specific binding platform. PMID- 19218568 TI - The C-terminal domain of the HIV-1 Vif protein is natively unfolded in its unbound state. AB - The human immunodeficiency virus type-1 (HIV-1) Vif protein neutralizes the cellular defense mechanism against the virus. The C-terminal domain of Vif (CTD, residues 141-192) mediates many of its interactions. Full-length Vif is difficult to purify in large amounts, hence the only available structure of Vif is of residues 140-155 within the ElonginBC complex. Other structural information, derived from modeling and indirect experiments, indicates that the Vif CTD may be unstructured. Here, we chemically synthesized the Vif CTD using pseudo-proline building blocks, studied its solution structure in the unbound state using biophysical techniques and found that it is unstructured under physiological conditions. The circular dichroism (CD) spectrum of Vif CTD showed a pattern of random coil with residual helical structure. The (15)N-HSQC nuclear magnetic resonance (NMR) spectrum was characteristic of natively unfolded peptides. Vif CTD eluted from an analytical gel filtration column earlier than expected, indicating an extended conformation. Disorder predictions found the CTD to be unstructured, in agreement with our experimental results. CD experiments showed that Vif CTD underwent a conformational change upon interacting with membrane mimicking DPC micelles, but not upon binding to a peptide derived from its binding region in ElonginC. Our results provide direct evidence for the unfolded structure of the free Vif CTD and indicate that it may gain structure upon binding its natural ligands. PMID- 19218566 TI - Krox20 controls the transcription of its various targets in the developing hindbrain according to multiple modes. AB - The zinc finger transcription factor Krox20 plays an essential role in the vertebrate hindbrain segmentation process. It positively or negatively controls a large variety of other regulatory genes, coordinating delimitation of segmental territories, specification of their identity, and maintenance of their integrity. We have investigated the molecular mechanisms of Krox20 transcriptional control by performing a detailed structure-function analysis of the protein in the developing chick hindbrain. This revealed an unsuspected diversity in the modes of action of a transcription factor in a single tissue, since regulation of each of the five tested target genes requires different parts of the protein and/or presumably different co-factors. The multiplicity of Krox20 functions might rely on this diversity. Investigation of known Krox20 co-factors was initiated in relation to this analysis. Nab was shown to act as a negative feedback modulator of the different Krox20 activating functions in the hindbrain. HCF-1 was found to bind to a Krox20 N-terminal region, which was shown to rely on multiple elements, including acidic domains, to convey Nab activation and Krox20 autoregulation. PMID- 19218569 TI - Medical management of pulmonary actinomycosis: data from 49 consecutive cases. PMID- 19218570 TI - Decreasing prevalence of levofloxacin-resistant Streptococcus pneumoniae in Hong Kong, 2001 to 2007. PMID- 19218571 TI - Association of high T allele frequency of CYP2B6 G516T polymorphism among ethnic south Indian HIV-infected patients with elevated plasma efavirenz and nevirapine. PMID- 19218572 TI - Functional relationship between bacterial cell density and the efficacy of antibiotics. AB - OBJECTIVES: To determine the functional relationship between the density of bacteria and the pharmacodynamics of antibiotics, and the potential consequences of this inoculum effect on the microbiological course of antibiotic treatment of Staphylococcus aureus infections. METHODS: In vitro time-kill, MIC estimation and antibiotic bioassay experiments were performed with S. aureus ATCC 25923 to ascertain the functional relationship between rates of kill and the MICs of six classes of antibiotics and the density of bacteria exposed. The potential consequences of the observed inoculum effects on the microbiological course of antibiotic treatment are explored with a mathematical model. RESULTS: Modest or substantial inoculum effects on efficacy were observed for all six antibiotics studied, such as density-dependent declines in the rate and extent of antibiotic mediated killing and increases in MIC. Although these measures of antibiotic efficacy declined with inoculum, this density effect did not increase monotonically. At higher densities, the rate of kill of ciprofloxacin and oxacillin declined with the antibiotic concentration. For daptomycin and vancomycin, much of this inoculum effect is due to density-dependent reductions in the effective concentration of the antibiotic. For the other four antibiotics, this density effect is primarily associated with a decrease in per-cell antibiotic concentration. With parameters in the range estimated, our mathematical model predicts that the course of antibiotic treatment can be affected by cell density; treatment protocols based on conventional (density independent) MICs can fail to clear higher density infections. CONCLUSIONS: The MICs used for pharmacokinetic/pharmacodynamic indices should be functions of the anticipated densities of the infecting population. PMID- 19218573 TI - Emergence of clonally related Klebsiella pneumoniae isolates of sequence type 258 producing plasmid-mediated KPC carbapenemase in Norway and Sweden. AB - BACKGROUND: The class A carbapenemase KPC has disseminated rapidly worldwide, challenging the treatment of Gram-negative infections. This report describes the first KPC-producing Klebsiella pneumoniae isolates identified in Norway (n=6) and the second isolate from Sweden. METHODS: Antimicrobial susceptibility profiles were determined using Etest. PCR and sequencing were used to determine the bla(KPC) variant, the surrounding genetic structure and the presence of AmpC and extended-spectrum beta-lactamase genes. PFGE and multilocus sequence typing (MLST) were used for epidemiological comparisons. Localization of bla(KPC) was investigated by S1 nuclease digestion, followed by PFGE and Southern blot hybridization. RESULTS: All isolates expressed a multidrug-resistant phenotype with some variability in the carbapenem susceptibility profile. The Norwegian isolates carried bla(KPC-2), while the Swedish isolate carried bla(KPC-3). All isolates carried TEM-1, but were negative for bla(CTX-M) and bla(AmpC) genes. SHV 11 and SHV-12 were detected in the Norwegian isolates, while the Swedish isolate carried only SHV-11. Isolates from four patients were associated with import from Greece (n=3) and Israel. The other isolates were probably associated with local transmissions. PFGE and MLST showed that the isolates were clonally related, with three isolates displaying ST258, a single locus variant of ST11 previously associated with the clonal spread of CTX-M-15-producing K. pneumoniae in Hungary. In all isolates, bla(KPC) was located on plasmids as part of isoform a of Tn4401. CONCLUSIONS: The emergence of KPC-producing isolates of K. pneumoniae in Norway and Sweden is associated with multiple import events and probable local transmission of a successful multiresistant ST258 clone, closely related to the CTX-M-15-producing ST11 clone previously described in Hungary. PMID- 19218574 TI - Phosphatidylinositol 3-kinase p85alpha regulatory subunit gene Met326Ile polymorphism in women with polycystic ovary syndrome. AB - BACKGROUND: Insulin resistance is a core feature of polycystic ovary syndrome (PCOS). Phosphatidylinositol (PI) 3-kinase is an important enzyme in the early insulin signaling cascade and plays a key role in insulin-mediated glucose transport. In its regulatory subunit, p85alpha, there is a common amino acid substitution (the Met326Ile polymorphism), and this amino acid may be crucial for the function of the p85alpha regulatory subunit and PI3-kinase. METHODS: Analysis of the Met326Ile polymorphism was carried out on DNA samples from 256 PCOS patients and 283 controls. Clinical and biochemical profiles of participants were also compared. RESULTS: The genotype distribution of the Met326Ile polymorphism in the PCOS group was not different from that of the controls (Met326Met/Met326Ile/Ile326Ile rates were 73.4%/23.4%/3.2% and 70.3%/26.1%/3.6% for the PCOS and control groups, respectively, P = 0.72). The PCOS group was divided into two subgroups according to the presence of the variant 326Ile allele. Compared with those carrying at least one variant 326Ile allele, carriers with the Met326Met genotype had higher serum 17-hydroxyprogesterone (17-OHP) {1.1 [95% confidence interval (CI) 1.1-1.3] ng/ml in those with the Met326Met genotype versus 0.8 (95% CI 0.7-1.0) ng/ml in those with Ile326Ile and Met326Ile genotypes, P = 0.0073} and free testosterone levels [1.2 (95% CI 1.1-1.4) pg/ml for Met326Met genotype versus 0.9 (95% CI 0.6-1.3) pg/ml for Ile326Ile and Met326Ile genotypes, P = 0.038]. CONCLUSIONS: Our results suggest that the PI3 kinase gene Met326Ile polymorphism may not be a major determinant for the development of PCOS, but it may modulate the concentrations of serum 17-OHP or free testosterone in PCOS patients. PMID- 19218575 TI - Single Day 2 embryo versus blastocyst-stage transfer: a prospective study integrating fresh and frozen embryo transfers. AB - BACKGROUND: Whether extended culture allowing selection of embryos with high development potential has any advantage over cleavage-stage embryo transfer remains a matter of debate. Among the currently unsolved questions, the cumulative delivery rate resulting from fresh and frozen embryo transfers needs to be taken into account in both strategies. The aim of our study was, therefore, to compare the efficacy of single embryo transfer either on Day 2 or on Day 5/6 combining fresh and frozen embryo transfers. METHODS: A prospective study including 478 couples assigned on a voluntary basis to undergo elective single embryo transfer (eSET, n = 243) on Day 2 or single blastocyst transfer (SBT, n = 235) on Day 5/6 was performed. The primary outcome measurement was the cumulative delivery rate including fresh and frozen-thawed cycles in both groups. RESULTS: The delivery rate per cycle following fresh embryo transfer was significantly higher in the SBT group compared with the eSET group (P < 0.01). Conversely, frozen embryo and/or blastocyst transfers tended to result in a higher number of deliveries in the eSET compared with the SBT group. Altogether, the cumulative delivery rate per couple, including fresh and frozen embryo transfers, was similar between the two groups (37.9% versus 34.2% in the SBT and eSET groups, respectively). CONCLUSIONS: The observed cumulative delivery rates in this study do not allow us to take a position in favor of SBT or eSET. An improvement in blastocyst cryopreservation may change this attitude. PMID- 19218576 TI - Remote implantable cardioverter defibrillator monitoring in a Brugada syndrome population. AB - AIMS: The diagnosis of Brugada syndrome (BS) is typically made in a young and otherwise healthy population. In patients with a high risk of sudden cardiac death (SCD), the only currently recommended therapy is an implantable cardioverter defibrillator (ICD), but these are not without complications. We investigated whether remote ICD monitoring could simplify follow-up and detect potential complications in these patients. METHODS AND RESULTS: Thirty-five consecutive patients (26 males, 44 +/- 11 years) implanted with an ICD for BS with a remote monitoring ['Home Monitoring' (HM), Biotronik, Germany] system were prospectively enrolled in this study. They were matched for age, sex, and follow up duration with 35 BS patients implanted with an ICD without this capability. During a mean follow-up of 33 +/- 17 months, the number of cardiology consultations was significantly lower in the HM group (3 +/- 2 vs. 7 +/- 3; P < 0.001). Inappropriate shock(s) [IS(s)] occurred in three patients (8.5%) in the HM group vs. six (17%) in the control group (P = NS). Ten patients in the HM group had a median of four alerts ('ventricular tachycardia/ventricular fibrillation detection' in all patients, 'shock' in three, 'ineffective shock' in two patients with shock on noise, 'extreme ventricular pacing impedance' in one patient due to lead failure, and 'deactivated therapy' in two patients with lead failure before replacement). In 5 of these 10 patients, prompt reprogramming of the ICD may have prevented IS(s). CONCLUSION: Remote ICD monitoring in patients with BS decreases outpatient consultations and may help prevent ISs. PMID- 19218577 TI - A global test of the pollination syndrome hypothesis. AB - BACKGROUND AND AIMS: 'Pollination syndromes' are suites of phenotypic traits hypothesized to reflect convergent adaptations of flowers for pollination by specific types of animals. They were first developed in the 1870s and honed during the mid 20th Century. In spite of this long history and their central role in organizing research on plant-pollinator interactions, the pollination syndromes have rarely been subjected to test. The syndromes were tested here by asking whether they successfully capture patterns of covariance of floral traits and predict the most common pollinators of flowers. METHODS: Flowers in six communities from three continents were scored for expression of floral traits used in published descriptions of the pollination syndromes, and simultaneously the pollinators of as many species as possible were characterized. KEY RESULTS: Ordination of flowers in a multivariate 'phenotype space' defined by the syndromes showed that almost no plant species fall within the discrete syndrome clusters. Furthermore, in approximately two-thirds of plant species, the most common pollinator could not be successfully predicted by assuming that each plant species belongs to the syndrome closest to it in phenotype space. CONCLUSIONS: The pollination syndrome hypothesis as usually articulated does not successfully describe the diversity of floral phenotypes or predict the pollinators of most plant species. Caution is suggested when using pollination syndromes for organizing floral diversity, or for inferring agents of floral adaptation. A fresh look at how traits of flowers and pollinators relate to visitation and pollen transfer is recommended, in order to determine whether axes can be identified that describe floral functional diversity more successfully than the traditional syndromes. PMID- 19218578 TI - The pollination of a self-incompatible, food-mimic orchid, Coelogyne fimbriata (Orchidaceae), by female Vespula wasps. AB - BACKGROUND AND AIMS: The study of specialized interactions between species is crucial to our understanding of processes in evolutionary ecology due to their profound effect on life cycles and diversification. Obligate pollination by a single wasp species is rare in Orchidaceae except in species with sexually deceptive flowers that are pollinated exclusively by male insects. The object of this study was to document pollination of the food-deceptive flowers of Coelogyne fimbriata, a species pollinated exclusively by female wasps. METHODS: Field observations and experiments were conducted in two populations of C. fimbriata. Floral phenology was recorded, and functional floral architecture was measured. Insect visitors to flowers were observed from 2005 to 2007. Bioassay experiments were conducted to check whether the floral odour attracted pollinators. Natural (insect-mediated) rates of pollinarium removal, pollinium deposition on stigmas, and fruit set were recorded. To determine the importance of cross-pollination, the breeding system was assessed via controlled, hand-pollination experiments. KEY RESULTS: Two populations of C. fimbriata with fragrant, nectarless flowers are pollinated by females of the same Vespula species (Vespidae, Hymenoptera). Experiments on wasps show that they crawl towards the source of the odour. The flowering period appears to coincide with an annual peak in Vespula colony expansion when additional workers forage for carbohydrates. Rates of pollinarium removal (0.069-0.918) and pollinium deposition on stigmas (0.025-0.695) are extremely variable. However, fruit set in C. fimbriata is always low (0.014 0.069) and appears to be based on self-incompatibility coupled with intraclonal (geitonogamous) deposition of pollinia. CONCLUSIONS: Coelogyne fimbriata and Steveniella satyrioides are now the only orchid species known to have food deceptive flowers that are pollinated exclusively by eusocial, worker wasps. In C. fimbriata, floral odour appears to be the major attractant. Sub-populations may go through flowering seasons when pollinators are abundant or infrequent, but fruit set always remains low because the obligate pollinator does not often appear to transfer pollinaria between intercompatible genets. PMID- 19218579 TI - Using phenotypic manipulations to study multivariate selection of floral trait associations. AB - BACKGROUND: A basic theme in the study of plant-pollinator interactions is that pollinators select not just for single floral traits, but for associations of traits. Responses of pollinators to sets of traits are inherent in the idea of pollinator syndromes. In its most extreme form, selection on a suite of traits can take the form of correlational selection, in which a response to one trait depends on the value of another, thereby favouring floral integration. Despite the importance of selection for combinations of traits in the evolution of flowers, evidence is relatively sparse and relies mostly on observational approaches. SCOPE: Here, methods for measuring selection on multivariate suites of floral traits are presented, and the studies to date are reviewed. It is argued that phenotypic manipulations present a powerful, but rarely used, approach to teasing apart the separate and combined effects of particular traits. The approach is illustrated with data from studies of alpine plants in Colorado and New Zealand, and recommendations are made about several features of the design of such experiments. CONCLUSIONS: Phenotypic manipulations of two or more traits in combination provide a direct way of testing for selection of floral trait associations. Such experiments will be particularly valuable if rooted in hypotheses about differences between types of pollinators and tied to a proposed evolutionary history. PMID- 19218580 TI - Effect of variation in self-incompatibility on pollen limitation and inbreeding depression in Flourensia cernua (Asteraceae) scrubs of contrasting density. AB - BACKGROUND AND AIMS: Selection may favour a partial or complete loss of self incompatibility (SI) if it increases the reproductive output of individuals in the presence of low mate availability. The reproductive output of individuals varying in their strength of SI may also be affected by population density via its affect on the spatial structuring and number of S-alleles in populations. Modifiers increasing levels of self-compatibility can be selected when self compatible individuals receive reproductive compensation by, for example, increasing seed set and/or when they become associated with high fitness genotypes. METHODS: The effect of variation in the strength of SI and scrub density (low versus high) on seed set, seed germination and inbreeding depression in seed germination (delta(germ)) was investigated in the partially self incompatible species Flourensia cernua by analysing data from self-, cross- and open-pollinated florets. KEY RESULTS: Examination of 100 plants in both high and low scrub densities revealed that 51% of plants were strongly self-incompatible and 49 % varied from being self-incompatible to self-compatible. Seed set after hand cross-pollination was higher than after open-pollination for self incompatible, partially self-incompatible and self-compatible plants but was uniformly low for strongly self-incompatible plants. Strongly self-incompatible and self-incompatible plants exhibited lower seed set, seed germination and multiplicative female fitness (floral display x seed set x seed germination) in open-pollinated florets compared with partially self-incompatible and self compatible plants. Scrub density also had an effect on seed set and inbreeding depression: in low-density scrubs seed set was higher after open-pollination and delta(germ) was lower. CONCLUSIONS: These data suggest that (a) plants suffered outcross pollen limitation, (b) female fitness in partially self-incompatible and self-compatible plants is enhanced by increased mate-compatibility and (c) plants in low-density scrubs received higher quality pollen via open-pollination than plants in high-density scrubs. PMID- 19218581 TI - Use of sediment CO2 by submersed rooted plants. AB - BACKGROUND AND AIMS: Submersed plants have different strategies to overcome inorganic carbon limitation. It is generally assumed that only small rosette species (isoetids) are able to utilize the high sediment CO(2) availability. The present study examined to what extent five species of submersed freshwater plants with different morphology and growth characteristics (Lobelia dortmanna, Lilaeopsis macloviana, Ludwigia repens, Vallisneria americana and Hydrocotyle verticillata) are able to support photosynthesis supplied by uptake of CO(2) from the sediment. METHODS: Gross photosynthesis was measured in two-compartment split chambers with low inorganic carbon availability in leaf compartments and variable CO(2) availability (0 to >8 mmol L(-1)) in root compartments. Photosynthetic rates based on root-supplied CO(2) were compared with maximum rates obtained at saturating leaf CO(2) availability, and (14)C experiments were conducted for two species to localize bottlenecks for utilization of sediment CO(2). KEY RESULTS: All species except Hydrocotyle were able to use sediment CO(2), however, with variable efficiency, and with the isoetid, Lobelia, as clearly the most effective and the elodeid, Ludwigia, as the least efficient. At a water column CO(2) concentration in equilibrium with air, Lobelia, Lilaeopsis and Vallisneria covered >75% of their CO(2) requirements by sediment uptake, and sediment CO(2) contributed substantially to photosynthesis at water CO(2) concentrations up to 1000 micromol L(-1). For all species except Ludwigia, the shoot to root ratio on an areal basis was the single factor best explaining variability in the importance of sediment CO(2). For Ludwigia, diffusion barriers limited uptake or transport from roots to stems and transport from stems to leaves. CONCLUSIONS: Submersed plants other than isoetids can utilize sediment CO(2), and small and medium sized elodeids with high root to shoot area in particular may benefit substantially from uptake of sediment CO(2) in low alkaline lakes. PMID- 19218582 TI - Terrestrial orchid conservation in the age of extinction. AB - BACKGROUND: Conservation through reserves alone is now considered unlikely to achieve protection of plant species necessary to mitigate direct losses of habitat and the pervasive impact of global climate change. Assisted translocation/migration represent new challenges in the face of climate change; species, particularly orchids, will need artificial assistance to migrate from hostile environments, across ecological barriers (alienated lands such as farmlands and built infrastructure) to new climatically buffered sites. The technology and science to underpin assisted migration concepts are in their infancy for plants in general, and orchids, with their high degree of rarity, represent a particularly challenging group for which these principles need to be developed. It is likely that orchids, more than any other plant family, will be in the front-line of species to suffer large-scale extinction events as a result of climate change. SCOPE: The South West Australian Floristic Region (SWAFR) is the only global biodiversity hotspot in Australia and represents an ideal test bed for development of orchid conservation principles. Orchids comprise 6 % of all threatened vascular plants in the SWAFR, with 76 out of the 407 species known for the region having a high level of conservation risk. The situation in the SWAFR is a portent of the global crisis in terrestrial orchid conservation, and it is a region where innovative conservation solutions will be required if the impending wave of extinction is to be averted. Major threatening processes are varied, and include land clearance, salinity, burning, weed encroachment, disease and pests. This is compounded by highly specialized pollinators (locally endemic native invertebrates) and, in the most threatened groups such as hammer orchids (Drakaea) and spider orchids (Caladenia), high levels of mycorrhizal specialization. Management and development of effective conservation strategies for SWAFR orchids require a wide range of integrated scientific approaches to mitigate impacts that directly influence ecological traits critical for survival. CONCLUSIONS: In response to threats to orchid species, integrated conservation approaches have been adopted (including ex situ and translocation principles) in the SWAFR with the result that a significant, multidisciplinary approach is under development to facilitate conservation of some of the most threatened taxa and build expertise to carry out assisted migration to new sites. Here the past two decades of orchid conservation research in the SWAFR and the role of research based approaches for managing effective orchid conservation in a global biodiversity hotspot are reviewed. PMID- 19218583 TI - Wind of change: new insights on the ecology and evolution of pollination and mating in wind-pollinated plants. AB - BACKGROUND: The rich literature that characterizes the field of pollination biology has focused largely on animal-pollinated plants. At least 10 % of angiosperms are wind pollinated, and this mode of pollination has evolved on multiple occasions among unrelated lineages, and hence this discrepancy in research interest is surprising. Here, the evolution and functional ecology of pollination and mating in wind-pollinated plants are discussed, a theoretical framework for modelling the selection of wind pollination is outlined, and pollen capture and the occurrence of pollen limitation in diverse wind-pollinated herbs are investigated experimentally. SCOPE AND CONCLUSIONS: Wind pollination may commonly evolve to provide reproductive assurance when pollinators are scarce. Evidence is presented that pollen limitation in wind-pollinated plants may not be as common as it is in animal-pollinated species. The studies of pollen capture in wind-pollinated herbs demonstrate that pollen transfer efficiency is not substantially lower than in animal-pollinated plants as is often assumed. These findings challenge the explanation that the evolution of few ovules in wind pollinated flowers is associated with low pollen loads. Floral and inflorescence architecture is crucial to pollination and mating because of the aerodynamics of wind pollination. Evidence is provided for the importance of plant height, floral position, and stamen and stigma characteristics in promoting effective pollen dispersal and capture. Finally, it is proposed that geitonogamous selfing may alleviate pollen limitation in many wind-pollinated plants with unisexual flowers. PMID- 19218584 TI - Pollinator visitation patterns strongly influence among-flower variation in selfing rate. AB - BACKGROUND AND AIMS: Adjacent flowers on Mimulus ringens floral displays often vary markedly in selfing rate. We hypothesized that this fine-scale variation in mating system reflects the tendency of bumble-bee pollinators to probe several flowers consecutively on multiflower displays. When a pollinator approaches a display, the first flower probed is likely to receive substantial outcross pollen. However, since pollen carryover in this species is limited, receipt of self pollen should increase rapidly for later flowers. Here the first direct experimental test of this hypothesis is described. METHODS: In order to link floral visitation sequences with selfing rates of individual flowers, replicate linear arrays were established, each composed of plants with unique genetic markers. This facilitated unambiguous assignment of paternity to all sampled progeny. A single wild bumble-bee was permitted to forage on each linear array, recording the order of floral visits on each display. Once fruits had matured, 120 fruits were harvested (four flowers from each of five floral displays in each of six arrays). Twenty-five seedlings from each fruit were genotyped and paternity was unambiguously assigned to all 3000 genotyped progeny. KEY RESULTS: The order of pollinator probes on Mimulus floral displays strongly and significantly influenced selfing rates of individual fruits. Mean selfing rates increased from 21 % for initial probes to 78 % for the fourth flower probed on each display. CONCLUSIONS: Striking among-flower differences in selfing rate result from increased deposition of geitonogamous (among-flower, within-display) self pollen as bumble-bees probe consecutive flowers on each floral display. The resulting heterogeneity in the genetic composition of sibships may influence seedling competition and the expression of inbreeding depression. PMID- 19218585 TI - Results of survey for assessing awareness level regarding radiological hazards of tobacco smoking. AB - Human consumption of tobacco is as old as human history. However, injurious health effects due to tobacco smoking may not be evident to the public at large. This article presents results of a questionnaire based on a survey carried out in the metropolitan city of Lahore of Pakistan with an aim to understand the awareness level of the general population about the radiological hazards associated with tobacco smoking. Some 3600 participants from different educational backgrounds from all segments of the society participated in this survey. Analysis of the data collected concluded that the awareness level of the representative participants regarding the radiological hazards associated with tobacco smoking was alarmingly poor. These results suggest that a nationwide mass media campaign may be launched by the government authorities in Health and Environment departments to enlighten the general public in this respect to avoid tobacco-smoking-associated health risks. PMID- 19218586 TI - Screening/health checks: can the patient only win? Whole body magnetic resonance imaging approaches to tumour screening. AB - One of the greatest challenges for the next decade is the fight against cancer disease. These activities must be based on three premises: Cancer prevention; Cancer diagnosis; Cancer treatment. PMID- 19218587 TI - Differential permeabilization effects of Ca2+ and valinomycin on the inner and outer mitochondrial membranes as revealed by proteomics analysis of proteins released from mitochondria. AB - It is well established that cytochrome c is released from mitochondria when the permeability transition (PT) of this organelle is induced by Ca2+. Our previous study showed that valinomycin also caused the release of cytochrome c from mitochondria but without inducing this PT (Shinohara, Y., Almofti, M. R., Yamamoto, T., Ishida, T., Kita, F., Kanzaki, H., Ohnishi, M., Yamashita, K., Shimizu, S., and Terada, H. (2002) Permeability transition-independent release of mitochondrial cytochrome c induced by valinomycin. Eur. J. Biochem. 269, 5224 5230). These results indicate that cytochrome c may be released from mitochondria with or without the induction of PT. In the present study, we examined the protein species released from valinomycin- and Ca2+-treated mitochondria by LC MS/MS analysis. As a result, the proteins located in the intermembrane space were found to be specifically released from valinomycin-treated mitochondria, whereas those in the intermembrane space and in the matrix were released from Ca2+ treated mitochondria. These results were confirmed by Western analysis. Furthermore to examine how the protein release occurred, we examined the correlation between the species of released proteins and those of the abundant proteins in mitochondria. Consequently most of the proteins released from mitochondria treated with either agent were highly expressed proteins in mitochondria, indicating that the release occurred not selectively but in a manner dependent on the concentration of the proteins. Based on these results, the permeabilization effects of Ca2+ and valinomycin on the inner and outer mitochondrial membranes are discussed. PMID- 19218588 TI - Aberrant DNA methylation status in human uterine leiomyoma. AB - Aberrant DNA methylation has been implicated in tumorigenesis. This study was undertaken to establish the genome-wide DNA methylation profile in uterine leiomyomas and to investigate whether DNA methylation status is altered in uterine leiomyomas. For this purpose, restriction landmark genomic scanning (RLGS) was performed on a paired sample of leiomyoma and adjacent normal myometrium. The RLGS profile revealed 29 aberrant methylation spots (10 methylated and 19 demethylated) in leiomyoma in comparison with myometrium. One of the differently methylated genomic loci was newly identified as GS20656 from the human genome sequence database. In 9 of the 10 paired samples, the DNA methylation levels of the first exon of GS20656 were significantly lower in leiomyoma than in myometrium, suggesting the existence of a genomic locus under epigenetic regulation in uterine leiomyomas. Unexpectedly, DNA methyltransferase 1 (DNMT1) and DNMT3a mRNA expression levels were higher in leiomyoma than in myometrium. These facts suggest that other epigenetic factors besides DNMT are involved in local changes of DNA methylation at genome loci. The present study indicates not only aberrant genome-wide DNA methylation status in uterine leiomyomas but also the existence of a genomic locus that is differently methylated between normal myometrium and uterine leiomyoma. PMID- 19218590 TI - Diseases in chicks and laying hens during the first 12 years after battery cages were banned in Switzerland. AB - Between 1992 and 2003, a period of 12 years after the definitive ban on battery cages in Switzerland, more than 10,000 replacement chicks and laying hens were examined postmortem. There was a significant decrease in the incidence of viral diseases, mostly due to a reduction in Marek's disease, but there was a marked increase in bacterial diseases, particularly since 1999, mainly due to colisepticaemia in young laying hens. There was a steady decrease in parasitic infections, but the incidence of non-infectious diseases varied from year to year, with no clear trends. There were no significant emerging diseases or economic losses in the alternative housing systems. Vaccination and hygiene were the most effective precautions against infections, and control strategies brought about a marked decline in notifiable diseases, especially for Salmonella Enteritidis. Fifteen years after the ban on battery cages in Switzerland, the health and egg production of laying hens is good. PMID- 19218591 TI - Injuries to Australian veterinarians working with horses. AB - Data from a health risks of Australian veterinarians (HRAV) study were reviewed to identify reported serious injuries incurred while working with horses and the factors associated with these injuries. Of the 2188 serious injuries reported in the HRAV study, 1583 (72.3 per cent) were associated with animals, and of these, 453 (28.6 per cent) involved horses. Most of them were sustained in stock or handling yards. Factors associated with an increased frequency of injury included activities such as suturing, wound care, tubing and drenching. The parts of the body most commonly injured were the head and face and the lower extremities. Fractures were the most common type of serious injury. The use of safety precautions at the time of the injury was reported by 70 per cent of those injured. PMID- 19218592 TI - Rhinosporidiosis in a dog native to the UK. PMID- 19218593 TI - Osteoporosis in goats associated with phosphorus and calcium deficiency. PMID- 19218594 TI - Feline parvovirus infection in an Asian palm civet (Paradoxurus hermaphroditus). PMID- 19218595 TI - Isolation of Mycoplasma capricolum subspecies capricolum from a dairy calf. PMID- 19218598 TI - Methods for dispatching backyard poultry. PMID- 19218599 TI - Apolipoprotein E polymorphism and expression in type 2 diabetic patients with nephropathy: clinicopathological correlation. AB - BACKGROUND: Diabetic nephropathy represents a heterogeneous group of renal pathologies that may be associated with genetic susceptibility. There have been clinical reports on the risk association of diabetic nephropathy with an apolipoprotein E (ApoE) exon 4 polymorphism although its correlations with renal histopathological changes have not been explored. METHODS: A total of 213 adult autopsies with type 2 diabetes and 111 non-diabetic control cases were analysed. Genomic DNA samples were obtained from spleen tissues. The ApoE genotype was determined by PCR-LDR analysis. Histopathological examination of kidney sections was performed in a subset of 51 diabetic and 111 control cases. ApoE protein expression in diabetic carriers with similar clinical status was examined by immunohistochemical staining. RESULTS: In type 2 diabetes, epsilon2 carriers (P = 0.04; odds ratio = 5.42; 95% CI: 1.10-26.8) and epsilon3/epsilon4 (P = 0.04; odds ratio = 22.5; 95% CI: 1.11-454.90) genotype carriers were more likely to have glomerular hypertrophy than were epsilon3/epsilon3 carriers. The epsilon2 carriers showed an increase in glomerular ApoE protein expression. A correlation between ApoE genotype and nodular glomerulosclerosis was not found. CONCLUSIONS: Our findings confirm the risk association of the ApoE polymorphism with diabetic nephropathy in clinical studies and is the first study demonstrating the correlations between ApoE genotypes, protein expression and structural changes in diabetic nephropathy. PMID- 19218600 TI - Replication of the recessive STBMS1 locus but with dominant inheritance. AB - PURPOSE: Strabismus is a common eye disorder with a prevalence of 1% to 4%. Comitant strabismus accounts for approximately 75% of all strabismus, yet more is known about the less common incomitant disorders. Comitant strabismus is at least partly inherited, but only one recessive genetic susceptibility locus, on chromosome 7p, has been identified in one family. The purpose of this study was to determine the frequency of STBMS1 as a cause of primary nonsyndromic comitant esotropia (PNCE). METHODS: Twelve families were recruited within the UK Hospital Eye Service as children attended for treatment of PNCE. All consenting persons were clinically assessed, and DNA was sampled. Chromosome 7 microsatellite markers were genotyped in all 12 families, and LOD scores were calculated under recessive and dominant models. RESULTS: One family was linked to STBMS1; in three, linkage was significantly excluded; and the remainder were uninformative. Twenty-six members from three generations of the linked family were analyzed further. Five family members were defined as affected; two had esotropia with an accommodative element; and three underwent strabismus surgery and appeared to have had an infantile/early-onset esotropia. A maximum LOD score of 3.21 was obtained under a dominant mode of inheritance; a recessive model gave an LOD score of 1.2. CONCLUSIONS: This study confirms that PNCE can result from sequence variants in an unknown gene at the STBMS1 locus. However, this locus accounts for only a proportion of cases, and other genetic loci remain to be identified. In contrast with the previously reported family, the pedigree described in this study is consistent with dominant rather than recessive inheritance at the STBMS1 locus. PMID- 19218601 TI - Analysis of mutations of the PITX2 transcription factor found in patients with Axenfeld-Rieger syndrome. AB - PURPOSE: To assess the effects of previously uncharacterized PITX2 missense mutations found in patients with Axenfeld-Rieger syndrome and to determine the functional roles of the C-terminal region of PITX2. METHODS: Recombinant PITX2 proteins were analyzed with the use of cellular immunofluorescence, electrophoretic mobility shift, reporter transactivation, and protein half-life assays in human trabecular meshwork cells. RESULTS: Two homeobox mutations, R43W and R90C, resulted in severely reduced DNA-binding and transcriptional activation despite normal nuclear localization. L105V, located C-terminal to the homeodomain, resulted in normal localization, reporter gene transactivation, and protein half-life, but with an altered mobility shift pattern of protein-DNA complexes. N108T, also located C-terminal to the homeodomain, resulted in an altered mobility shift pattern and with slightly increased reporter transactivation and shortened protein half-life. The PITX2 C-terminal region contains at least three domains, each with distinct modulating effects on reporter transactivation. CONCLUSIONS: PITX2 homeobox mutations predictably resulted in decreased function of the protein. However, the two C-terminal mutations exhibited only subtle defects on PITX2 transactivation and protein-DNA binding, suggesting that ocular development is sensitive to even slight alterations of PITX2 function. The C-terminal mutations L105V and N108T lie in a domain that inhibits PITX2 transcriptional activation. These two mutations produce electrophoretic mobility shift assay patterns representing altered protein-DNA interactions that may be important for accurate target gene selection. Additionally, N108T resulted in a less stable PITX2 mutant protein with elevated activity that may result in stochastic dysregulation during critical stages of development. Together, the results clearly indicate that stringent control of PITX2 is required for normal ocular development and function. PMID- 19218602 TI - Outflow facility in isolated porcine eyes after creation of an intrascleral canal by injection of stabilized hyaluronic acid. AB - PURPOSE: To investigate outflow facility in isolated porcine eyes after creation of an intrascleral canal by injection of stabilized, nonanimal, hyaluronic acid gel (NASHA; Q-Med AB, Uppsala, Sweden). METHODS: Outflow facility of 60 porcine eyes was measured after creation of an intrascleral canal into the anterior chamber by injection of NASHA gel using six different combinations of needle size (21, 23, and 27 gauge) and canal length (4.6 mm). Ten eyes were tested in each of the six subgroups and an additional 10 were control eyes. After cannulation of the anterior chamber and adjustment of the inflow rate to achieve a stable pressure of 10 mm Hg, an intrascleral channel was created by injection of NASHA gel. The inflow rate was then increased to achieve a stable pressure of 20 mm Hg and then again adjusted to maintain a stable pressure of 30 mm Hg. RESULTS: A significant increase in outflow facility was found between the control group and the NASHA gel-injected group (60 eyes) for both the 10- to 20-mm Hg and the 20- to 30-mm Hg pressure change (P < 0.001). Increase in outflow facility was independent of the canal lengths and the needle sizes used for both the 10- to 20 mm Hg (P = 0.82) and 20- to 30-mm Hg (P = 0.99) pressure change. CONCLUSIONS: A single ab externo intrascleral canal created by injection of NASHA gel increases the outflow facility in isolated porcine eyes. This is a potentially promising new technique for lowering intraocular pressure. It remains to be seen whether these positive results can be reproduced for significant periods in humans. PMID- 19218603 TI - Genetic dissociation of dacryoadenitis and sialadenitis in a Sjogren's syndrome mouse model with common and different susceptibility gene loci. AB - PURPOSE: Sjogren's syndrome (SS) is a systemic autoimmune disease in which the main lesions are dacryoadenitis and sialadenitis. It is unclear whether these lesions develop in a common genetic background. A quantitative trait locus (QTL) analysis was performed in the SS mouse model, MRL/MpJ-lpr/lpr (MRL/lpr), to identify the susceptibility loci to dacryoadenitis and sialadenitis and the association with both loci. METHODS: MRL/lpr, C3H/HeJ-lpr/lpr (C3H/lpr), (MRL/lpr x C3H/lpr) F1, and (MRL/lpr x C3H/lpr) F2 intercross mice were prepared, and the severity of dacryoadenitis and sialadenitis in individuals was quantified by histopathologic grading. In genomic DNA samples from the F2 mice, the polymorphic microsatellite markers highly associated with each lesion were determined as susceptibility loci. RESULTS: QTLs with significant linkage for dacryoadenitis were mapped on chromosome 1 (the position of maximum logarithm of odds [LOD] score; 64.1 cM), designated Adacm1; chromosome 2 (88.4 cM), Adacm2; and chromosome 5 (63.9 cM), Adacm3. Those for sialadenitis were mapped on chromosome 1 (69.0 cM), Asm3, and chromosome 2 (65.3 cM and 82.1 cM), Asm4 and Asm5. Adacm1/Asm3 and Adacm2/Asm5 seemed to be a common chromosomal region, respectively. MRL-homozygous at Adacm1 and Adacm2 and at Asm3 and Asm5 manifested an additive effect on the development of dacryoadenitis and sialadenitis, respectively, whereas Adacm3 did not. CONCLUSIONS: Dacryoadenitis and sialadenitis in MRL/lpr mice are under the control of common and different susceptibility loci, with an allelic combination that leads to regular variations in pathologic phenotypes. PMID- 19218604 TI - AlphaB-crystallin: a Golgi-associated membrane protein in the developing ocular lens. AB - PURPOSE: All crystallins have non-crystallin catalytic functions. Because catalytic functions do not require large concentrations of protein, as are seen in the lens, there is a perception of dichotomy in the catalytic/physiological function of crystallins within and outside the lens. The status of alphaB crystallin, a ubiquitously expressed small heat shock protein (and a crystallin) in the ocular lens, was investigated. METHODS: Discontinuous sucrose density gradients were used for fractionation of Golgi membranes and vesicles. Light microscopy and confocal microscopy were used for immunolocalization in cultured cells and the native lens. RESULTS: alphaB-crystallin is highly organized, as indicated by its polar presence in the apical Golgi in lens epithelium and in the perinuclear Golgi streaks in differentiating lens fiber cells. Assessment of the distribution of alphaB-crystallin in Golgi-enriched and vesicular fractions (characterized by the presence of Golgi membrane protein GM130 and vesicle coat protein gammaCOP) in the developing lens reveal a gradual transition from Golgi to vesicular fraction, concomitant with the appearance of alphaB-crystallin as a "soluble" protein. CONCLUSIONS: These data demonstrate that alphaB-crystallin, known to be a soluble protein, starts life as a Golgi-associated membrane protein in the fetal and early postnatal lens and that the developmentally controlled physical state of the Golgi determines the status of this protein in the lens. These findings also show the similarity in the localization/physiological function of alphaB-crystallin within and outside the ocular lens and suggest that non-crystallin/catalytic function is an innate component of the expression of a crystallin in the lens. PMID- 19218605 TI - Route of penetration of topically instilled nipradilol into the ipsilateral posterior retina. AB - PURPOSE: To investigate how topically instilled nipradilol penetrates the ipsilateral posterior retina-choroid in normal rabbit eyes. METHODS: Albino rabbits were used. Topical instillation (1%, 100 microL) or intracameral (0.1%, 100 microL) or sub-Tenon injection (0.1%, 10 microL) of [(14)C]nipradilol was performed in one eye. Ocular and periocular distribution and the concentrations of [(14)C]nipradilol were determined by whole-head autoradiography, the results of which were validated by measurements in isolated tissues. In addition, the unchanged form of nonradiolabeled nipradilol in the posterior retina after topical instillation (1%, 100 microL) was quantified by liquid chromatography tandem mass spectrometry (LC/MS/MS). RESULTS: Autoradiography revealed that the nipradilol concentration after topical instillation was higher in the ipsilateral posterior retina-choroid than on the contralateral side (142.9 ng/g vs. 108.3 ng/g, P = 0.026), and in the periocular tissue around the optic nerve insertion on the ipsilateral side than on the contralateral side (207.1 ng/g vs. 141.1 ng/g, P < 0.001). After intracameral injection, radioactivity was observed only in anterior, but not posterior parts of the eye. Radioactivity was observed only in the ipsilateral posterior retina-choroid and periocular tissues around the optic nerve insertion after sub-Tenon injection. The results in the isolated tissues validated autoradiographic evaluations. CONCLUSIONS: These results suggest that diffusion from posterior periocular tissues across the posterior sclera may be the main route for local penetration of the instilled drug to reach the posterior retina-choroid in albino rabbits. PMID- 19218606 TI - Neurologic evaluation of acute lacrimomimetic effect of cyclosporine in an experimental rabbit dry eye model. AB - PURPOSE: To evaluate neurologically acute lacrimation caused by cyclosporine (CsA) eyedrops in rabbit. METHODS: Normal adult male New Zealand White rabbits and those that underwent parasympathectomy each received a single instillation of 0.1% CsA or vehicle eyedrops. Schirmer tear test (STT) results, flow rate of lacrimal gland (LG) fluid from the excretory lacrimal duct of the main LG, and blink rate (over a 3-minute period) were measured before and after instillation of CsA or vehicle. Light microscopy was performed to examine the main LG in vitro. Protein release from LG fragments was assessed after incubation with CsA for 30 minutes. RESULTS: In normal rabbits, the STT value and the flow rate of LG fluid were significantly increased after treatment with CsA compared with vehicle (P < 0.05). In contrast, no changes were found in denervated eyes. The blink rate of CsA-treated eyes was significantly higher than that of vehicle-treated eyes in normal rabbits (P < 0.005), whereas that of denervated eyes decreased significantly after CsA instillation compared with before administration (P < 0.005). Light microscopy showed that the cytoplasm of acinar cells was packed with secretory granules in denervated LG tissue 7 days after parasympathectomy. The same finding was observed 3 hours after CsA instillation. CsA had no stimulatory effect on protein release by acinar cells in LG fragments at all concentrations tested. CONCLUSIONS: These results suggest that CsA has no direct effect on tear fluid secretion from the LG in an acute model. Instead, CsA increases reflex tear flow. PMID- 19218607 TI - Corneal endothelial cell proliferation: a function of cell density. AB - PURPOSE: To determine how stimuli that increase corneal endothelial cell proliferation of human corneas in culture relate to changes in endothelial cell density in the central and peripheral cornea. METHODS: Human donor cadaver corneas not suitable for transplantation were divided into four pie-shaped wedges and incubated at 37 degrees C in medium supplemented with fetal bovine serum, epidermal growth factor, fibroblast growth factor, and gentamicin. To promote a proliferative response, samples were treated with EDTA at concentrations of 0, 0.5, 2.5, and 5.0 mM for 1 hour and then returned to culture medium. Endothelial cell proliferation was assayed with Ki-67 immunolocalization 48 and 96 hours after EDTA treatment. Samples were mounted with propidium iodide or DAPI. The total number of cells and the number of Ki-67-positive cells were counted in three regions, defined as central, mid, and peripheral cornea, to determine endothelial cell density and percentage of proliferation. RESULTS: A proliferative response to EDTA was not found. However, increased proliferation was noted in the central compared with the peripheral corneal region. Unexpectedly, the increased proliferation in the central region corresponded to a trend of lower endothelial cell density in the central region compared with the peripheral region. Corneal endothelial cell proliferation under our culture conditions is noted primarily when cell density is less than 2000 cells/mm(2). CONCLUSIONS: Corneal endothelial cell proliferation under our culture conditions does not lead to supranormal endothelial cell density. Rather, cell proliferation is noted in those regions that may be experiencing a greater burden of cell loss. PMID- 19218608 TI - Role of adaptive immunity in the pathogenesis of Candida albicans keratitis. AB - PURPOSE: Innate immunity had been thought to be critical in the pathogenesis and prognosis of fungal keratitis. This study was conducted to determine whether experimental Candida albicans keratitis (CaK) induces an adaptive immune response. METHODS: Experimental murine CaK was induced by intrastromal injection of C. albicans spores, and fungal pneumonia was induced by intranasal inhalation of spores. Active immunization was accomplished by subcutaneous injection of heat inactivated spores. Serum was collected at different times after the induction of primary or secondary CaK for the measurement of IgA, IgG, IL-4, and IFN-gamma. Immunohistochemistry was used to detect immunoglobulin deposition and lymphocyte infiltration in diseased corneas. RESULTS: After intrastromal injection of C. albicans spores in immunocompetent mice, typical CaK occurred, and the corneas healed in 3 weeks. When recovered corneas were challenged again with spores, they developed milder CaK and healed faster than with primary CaK. Mice that had recovered from pulmonary infection or had been immunized also showed increased resistance to CaK. Compared with naive mice, the mice that had previously encountered C. albicans produced more IgG and IgA in serum and more immunoglobulin deposition and lymphocyte infiltration in corneas on secondary CaK induction. Cytokines assays showed that the immune response induced by CaK was biased toward the T-helper (Th)1 type. CONCLUSIONS: Th1-type adaptive immune response and immunologic memory were induced by C. albicans keratitis, and previous contact with Candida preparation enhanced the resistance of the host to subsequent corneal challenge with the same fungus. Active immunization might be an effective strategy to prevent fungal keratitis in populations at high risk. PMID- 19218609 TI - Upper and lower tear menisci in the diagnosis of dry eye. AB - PURPOSE: To measure the upper and lower tear menisci in patients with aqueous tear deficiency (ATD) dry eye by real-time optical coherence tomography (OCT) and to determine the most effective meniscus variables for the diagnosis of dry eye. METHODS: Eyes of 48 pre-screened ATD patients were compared with those of 47 healthy subjects. Upper and lower tear menisci were imaged simultaneously by real time OCT immediately after blinking. The height, radius, and cross-sectional area of upper and lower tear menisci were obtained. RESULTS: The tear meniscus radius, height, and cross-sectional area were significantly smaller in patients with dry eye than in healthy subjects (P < 0.01). The lower tear meniscus variables were higher than the upper menisci (P < 0.01) in normal subjects; however, no significant differences between menisci were found in ATD patients. In both groups, the upper and lower tear meniscus variables were strongly correlated with each other. Good dry eye diagnostic accuracies were obtained with cutoff values for an abnormal lower tear meniscus radius (LTMR) of 182 microm and a lower tear meniscus height (LTMH) of 164 microm. The LTMR diagnostic sensitivity and specificity were 0.92 and 0.87, respectively. The LTMH diagnostic sensitivity and specificity were 0.92 and 0.90. CONCLUSIONS: Upper tear meniscus variables in ATD patients were assessed by real-time OCT. The tear meniscus was smaller in ATD patients than in healthy subjects. LTMR and LTMH may have potential in the diagnosis of ATD. PMID- 19218610 TI - Impact of cell source on human cornea reconstructed by tissue engineering. AB - PURPOSE: To investigate the effect of the tissue origin of stromal fibroblasts and epithelial cells on reconstructed corneas in vitro. METHODS: Four types of constructs were produced by the self-assembly approach using the following combinations of human cells: corneal fibroblasts/corneal epithelial cells, corneal fibroblasts/skin epithelial cells, skin fibroblasts/corneal epithelial cells, skin fibroblasts/skin epithelial cells. Fibroblasts were cultured with ascorbic acid to produce stromal sheets on which epithelial cells were cultured. After 2 weeks at the air-liquid interface, the reconstructed tissues were photographed, absorption spectra were measured, and tissues were fixed for histologic analysis. Cytokine expression in corneal- or skin-fibroblast conditioned media was determined with the use of protein array membranes. The effect of culturing reconstructed tissues with conditioned media, or media supplemented with a cytokine secreted mainly by corneal fibroblasts, was determined. RESULTS: The tissue source from which epithelial and mesenchymal cells were isolated had a great impact on the macroscopic and histologic features (epithelium thickness and differentiation) and the functional properties (transparency) of the reconstructed tissues. The reconstructed cornea had ultraviolet-absorption characteristics resembling those of native human cornea. The regulation of epithelial differentiation and thickness was mesenchyme dependent and mediated by diffusible factors. IL-6, which is secreted in greater amounts by corneal fibroblasts than skin fibroblasts, decreased the expression of the differentiation marker DLK in the reconstructed epidermis. CONCLUSIONS: The tissue origin of fibroblasts and epithelial cells plays a significant role in the properties of the reconstructed tissues. These human models are promising tools for gaining a thorough understanding of epithelial-stromal interactions and regulation of epithelia homeostasis. PMID- 19218611 TI - The contribution of lipid layer movement to tear film thinning and breakup. AB - PURPOSE: To investigate whether the tear film thinning between blinks is caused by evaporation or by tangential flow of the tear film along the surface of the cornea. Tangential flow was studied by measuring the movement of the lipid layer. METHODS: Four video recordings of the lipid layer of the tear film were made from 16 normal subjects, with the subjects keeping their eyes open for up to 30 seconds after a blink. To assess vertical and horizontal stretching of the lipid layer and underlying aqueous layer, lipid movement was analyzed at five positions, a middle position 1 mm below the corneal center, and four positions respectively 1 mm above, below, nasal, and temporal to this middle position. In addition, in 13 subjects, the thinning of the tear film after a blink was measured. RESULTS: The total upward movement could be fitted by the sum of an exponential decay plus a slow steady drift; this drift was upward in 14 of 16 subjects (P = 0.002). Areas of thick lipid were seen to expand causing upward or downward drift or horizontal movement. The velocity of the initial rapid upward movement and the time constant of upward movement were found to correlate significantly with tear film thickness but not with tear-thinning rate. CONCLUSIONS: Analysis indicated that the observed movement of the lipid layer was too slow to explain the observed thinning rate of the tear film. In the Appendix, it is shown that flow under a stationary lipid layer cannot explain the observed thinning rate. It is concluded that most of the observed tear thinning between blinks is due to evaporation. PMID- 19218612 TI - Long-term culture and growth kinetics of murine corneal epithelial cells expanded from single corneas. AB - PURPOSE: To develop a reproducible procedure for the long-term culture of corneal epithelial cells from a single mouse cornea. METHODS: Corneal limbal explants of C57BL6/J mice were cultured in serum-free, low-Ca(2+) medium supplemented with EGF and cholera toxin. Epithelial cells were subcultured at a 1:3 split until passage (P)4 and at lower densities after P4. Colony-forming efficiency, population-doubling times, and population doublings were determined. The expression of p63, keratin (K)19, K12, and involucrin was analyzed by RT-PCR, immunocytochemistry, and Western blotting. Differentiation potential was examined by switching the medium to serum or high Ca(2+)-containing medium. Stratification ability was analyzed by air-lift culture. RESULTS: Thirty of 32 (93.8%) corneal explants were successfully subcultured to P1. Cultures without cholera toxin did not proliferate past P2 (n = 12), but 55% of cultures supplemented with cholera toxin achieved P4 (n = 20). After P4, cells were stably subcultured over 25 passages. Colony-forming efficiency increased from 9.7% +/- 2.6% at P5 to 29.0% +/- 3.3% at P20. The cells showed cobblestone appearance and expressed p63, K19, and involucrin but were negative for K12. Serum and high Ca(2+) induced differentiation, and cells cultured in DMEM/F12 with serum showed K12 mRNA expression. Stratified epithelium was formed by air-lifting. CONCLUSIONS: With this procedure, corneal epithelial cells from a single cornea can be cultured long term and can retain the potential to differentiate and stratify. This procedure can be a powerful tool for studies that require comparison of corneal epithelial cells from normal and transgenic mice in vitro. PMID- 19218613 TI - Detailed ophthalmologic evaluation of 43 individuals with PAX6 mutations. AB - PURPOSE: Heterozygous mutations of the PAX6 gene cause a variety of ocular malformations, the best known being aniridia (absence of the iris). Mutation analyses and detailed clinical evaluations were performed in 43 individuals with aniridia or closely related ocular anomalies, to investigate whether phenotype correlates with mutation type. METHODS: Case notes and medical records were reviewed and patients were reexamined when necessary. Denaturing high-performance liquid chromatography (DHPLC) analysis and sequencing of the PAX6 coding region was performed in individuals whose mutation was unknown. RESULTS: The most common PAX6 mutations identified were premature termination mutations, amino acid substitutions, and C-terminal extensions. Six novel mutations are reported. Mutations that inactivate one copy of the gene typically caused a severe phenotype including foveal hypoplasia, marked iris anomalies, and severe visual impairment. Missense mutations, all affecting invariant amino acids in the paired domain, caused milder phenotypes in this cohort, with a lower incidence of foveal hypoplasia and less severe visual loss. C-terminal extension mutations caused relatively severe anomalies and marked reduction in vision. Two C-terminal extension cases had a unilateral exudative retinopathy, resembling Coats' disease, which has not previously been reported in association with PAX6 mutation. CONCLUSIONS: PAX6 mutations cause panocular malformations that vary considerably in pattern and severity. In our cohort, iris hypoplasia, nystagmus, and foveal hypoplasia were most common, with cataracts, corneal anomalies, and high refractive errors also frequently observed. In this cohort, loss-of-function and C-terminal extension mutations were found to cause more severe phenotypes than missense mutations. PMID- 19218614 TI - Concentration and bioavailability of ciprofloxacin and teicoplanin in the cornea. AB - PURPOSE: To investigate the concentration and bioavailability of ciprofloxacin and teicoplanin in the cornea. METHODS: A biological assay was developed with corneal tissue used as a carrier for the antimicrobial. Concentration and biological activity were determined with a chemical assay and zone of inhibition (ZOI) around corneal samples with epithelial and endothelial surfaces in contact with the indicator organism. Patients undergoing penetrating keratoplasty received ciprofloxacin 0.3% or teicoplanin 1%. RESULTS: There were good correlations between antimicrobial concentration and ZOI, when either filter paper or corneal discs were used (R(2) > 92%). Of 33 patients, the mean (median) concentration of ciprofloxacin in the cornea was 1.37 mg/L (0.46 mg/L) and 1.89 mg/L (1.44 mg/L; bioassay) in the epithelial and endothelial orientations, respectively, and 14.87 mg/L (7.41) in the cornea and 0.51 mg/L (0.42) in the aqueous (chemical assay). For teicoplanin, the mean (median) concentration in the cornea was 9.58 mg/L (0 mg/L) in the epithelial and 4.78 mg/L (0 mg/L) in the endothelial orientations (bioassay). In the chemical assay, teicoplanin could not be detected in the cornea or aqueous at the lower limit of detection of 3.6 mg/L. CONCLUSIONS: The ZOI produced by corneal tissue provides a potential bioassay of antimicrobial activity and concentration. Although in contrast to teicoplanin ciprofloxacin shows good corneal penetration, with high endothelial-to-epithelial levels, only approximately 10% of measured levels in a chemical assay are available, according to a bioassay. Teicoplanin shows relatively poor corneal penetration through intact epithelium. These methods may be useful in evaluating the biological activity across the cornea of antimicrobials introduced into ophthalmic practice to deal with changing bacterial resistance. PMID- 19218615 TI - Early synaptic defects in tulp1-/- mice. AB - PURPOSE: Mutations in the photoreceptor-specific tubby-like protein 1 (TULP1) underlie a form of autosomal recessive retinitis pigmentosa. To investigate the role of Tulp1 in the photoreceptor synapse, the authors examined the presynaptic and postsynaptic architecture and retinal function in tulp1(-/-) mice METHODS: The authors used immunohistochemistry to examine tulp1(-/-) mice before retinal degeneration and made comparisons with wild-type (wt) littermates and retinal degeneration 10 (rd10) mice, another model of photoreceptor degeneration that has a comparable rate of degeneration. Retinal function was characterized with the use of electroretinography. RESULTS: In wt mice, Tulp1 is localized to the photoreceptor synapse. In the tulp1(-/-) synapse, the spatial relationship between the ribbon-associated proteins Bassoon and Piccolo are disrupted, and few intact ribbons are present. Furthermore, bipolar cell dendrites are stunted. Comparable abnormalities are not seen in rd10 mice. The leading edge of the a wave had normal kinetics in tulp1(-/-) mice but reduced gain in rd10 mice. The b wave intensity-response functions of tulp1(-/-) mice are shifted to higher intensities than in wt mice, but those of rd10 mice are not. CONCLUSIONS: Photoreceptor synapses and bipolar cell dendrites in tulp1(-/-) mice display abnormal structure and function. A malformation of the photoreceptor synaptic ribbon is likely the cause of the dystrophy in bipolar cell dendrites. The association of early-onset, severe photoreceptor degeneration preceded by synaptic abnormalities appears to represent a phenotype not previously described. Not only is Tulp1 critical for photoreceptor function and survival, it is essential for the proper development of the photoreceptor synapse. PMID- 19218617 TI - Thyroid hormone induces a time-dependent opsin switch in the retina of salmonid fishes. AB - PURPOSE: To determine the role of thyroid hormone in inducing the UV (SWS1)-to blue (SWS2) opsin switch in the retina of two salmonid fishes, the coho salmon (Oncorhynchus kisutch) and the rainbow trout (O. mykiss). METHODS: Fish were treated with thyroid hormone (T(4)) or the vehicle solution (0.1 M NaOH, control), exogenously or by intraocular injection, at different life history stages. Microspectrophotometry and in situ hybridization with riboprobes against the SWS1 and SWS2 opsins were used to reveal the dynamics of opsin expression in treated and control animals. To assess whether thyroid hormone induced differentiation of retinal progenitor cells into cones, treated and control fish were injected intraocularly with bromodeoxyuridine (BrdU) and the number of proliferating cells in the outer nuclear layer (ONL) determined. These observations were accompanied by histologic counts of cone densities. RESULTS: Thyroid hormone induced a reversible UV-to-blue opsin switch in differentiated single cones of juvenile salmonids (alevin and parr stages), but failed to exert any effect in the retina of older fish (smolt stage). The switch progressed from the ventral to the dorsal retina in clockwise fashion. Thyroid hormone did not induce cone density changes or alterations in the number of BrdU-labeled cells, which were the same in control and treated animals. CONCLUSIONS: Thyroid hormone induces a UV (SWS1)-to-blue (SWS2) opsin switch in the retina of young salmonid fishes that is identical with that occurring during natural development. The switch occurs in differentiated photoreceptors, is reversible (maintained by thyroid hormone exposure), and can be induced only before its natural onset. Thyroid hormone did not cause changes in the number of proliferating cells in the ONL. These results conform to the dynamics of thyroid hormone-induced opsin expression in the mouse and are consistent with the opsin plasticity found in differentiated photoreceptors of the fruit fly, Drosophila melanogaster. This work establishes a role for thyroid hormone in triggering opsin switches in the vertebrate retina. PMID- 19218616 TI - Regulatory sequences in the 3' untranslated region of the human cGMP phosphodiesterase beta-subunit gene. AB - PURPOSE: Rod cGMP-phosphodiesterase, a key enzyme in visual transduction, is important for retinal integrity and function. Mutations in the gene encoding the phosphodiesterase beta-subunit (PDEbeta) cause retinal degeneration in animals and humans. Here the authors tested the hypothesis that elements in the 3' untranslated region (3' UTR) of the PDEbeta gene are involved in the regulation of PDEbeta expression. METHODS: Involvement of the 3' UTR of PDEbeta mRNA in the regulation of PDEbeta expression was assessed by Y-79 retinoblastoma cells or the heads of Xenopus laevis tadpoles with constructs containing the SV40 or PDEbeta promoter, the luciferase cDNA, and either the SV40 or the PDEbeta 3' UTR (or fragments of its sequence). RESULTS: Compared with the SV40 3' UTR (used as control), the entire PDEbeta 3' UTR decreased reporter gene expression in Y-79 retinoblastoma cells as well as in SY5Y neuroblastoma and 293 human embryonic kidney cell lines. However, the authors observed that two 100-nucleotide fragments from the PDEbeta 3' UTR increased while its noncanonical poly adenylation signal abolished reporter gene expression in Y-79 retinoblastoma cells and in ex vivo experiments using Xenopus tadpole heads. In particular, an 11-nucleotide element (EURE) in one of the 100-nucleotide fragments was responsible for the upregulation of luciferase expression. CONCLUSIONS: These studies indicate that the 3' UTR of the PDEbeta mRNA is involved in the complex regulation of this gene's expression in the retina. Moreover, the results show that the PDEbeta poly-A signal has a dominant inhibitory effect over two other regions in the 3' UTR that stimulate gene expression. PMID- 19218618 TI - Intersession repeatability of contrast sensitivity scores in age-related macular degeneration. AB - PURPOSE: To describe the intersession repeatability of contrast sensitivity (CS) measurement using Pelli-Robson charts in patients with age-related macular degeneration. METHODS: Repeatability was calculated from three measurements of CS over a 12-week period using a standardized protocol in 107 nontreated eyes of 107 patients with age-related macular degeneration who were enrolled in an ongoing clinical trial. RESULTS: Data from 91 patients were included in the analysis, with a 95% coefficient of repeatability of 7 letters (0.35 log CS), ranging from 6 letters for 32 eyes with drusen only to 8 letters for patients with late AMD (macular scars or geographic atrophy). Three (3%) of these stable patients had an apparent six or more letter reduction in contrast sensitivity at the week 1 visit compared with baseline. CONCLUSIONS: There is a high intersession test-retest variability of Pelli-Robson CS scores in patients with AMD, with implications for AMD clinical trial design. Although a change criterion of six or more letters may be an adequate end point in clinical trials for patients with early AMD, a larger change criterion may be necessary for clinical trials of patients with late AMD. PMID- 19218619 TI - The Journal of Physiology Annual Report 2008. PMID- 19218620 TI - Reporting ethical matters in the Journal of Physiology: standards and advice. AB - Reporting of ethical matters in The Journal is very important. To advise and assist authors, particularly those who may be less familiar with the legislation in the UK, this article sets out the basic principles and methods that should be used and provides many key web sources of information. It addresses the structure of regulations, and introduces the concept of research governance. The UK law is summarized. Advice is given on the format and description of experiments, and common problems addressed. Aspects of human studies are addressed. Ethical considerations of publication such as authorship and originality, and problems such as plagiarism and fabrication are described. Updates will be published regularly. PMID- 19218621 TI - Measuring synthesis rates of different proteins: clues to training adaptations. PMID- 19218622 TI - Role of hippocampal NMDA receptors in a mouse model for fragile X mental retardation syndrome. PMID- 19218623 TI - [Recent crisis of psychiatry in the context of modern and postmodern science]. AB - INTRODUCTION: We have investigated the recent crisis of psychiatry in perspective of the last two centuries of its history. First and longer part of this period belongs to the Modern era. The second part beginning in the sixties of the last century is the Postmodern era, the one we live in today. We have pointed out that the recent crisis has not come by accident but as one of the several changes characterising our transition into the era labelled "Postmodern". METHODS: The crisis of psychiatry was analysed in respect of the development of science and in social context. We focused on psychiatry and science and their interconnectedness in their historical articulations. A way out of the crisis of the current era can be found also with the use of historical methods. RESULTS: Science is changing both in respect of its quality and its role in our age. Taking into consideration normal and post-normal science, we differentiated two additional generations of science, pro-normal science and perato-scientia, in the most recent history. On the one hand, psychiatry serves as a paradigm for the conceptualisation of contemporary science. On the other hand, as an up-to-date science re conceptualised partly according to its own paradigm it may find a way out of its own crisis. The many facets of the current crisis were demonstrated by analysing recent developments of the Hungarian health politics. Concerning this topic we adopted the term "economicity" elaborated by us earlier. We found that psychiatry operated by the hegemony of the rules of economicity might become a relay station for selecting patients to be thrown away as human garbage. CONCLUSIONS: This catastrophic outcome may occur if a political system is organized purely by economicity rules without either historical responsibility or local solidarity. However, up-to-date trends of scientificity as shown by pro-normal science and perato-scientia guarantee a radically different course for psychiatry. Following this course, which is consistent with the insight into the common lot of normal and abnormal people, psychiatry should be guided also in the future by the categorical imperative of therapy. PMID- 19218624 TI - [Violent behaviour and stigmatisation among psychiatric patients]. AB - The prejudices existing in public opinion, such as mentally disordered people are dangerous, violent and unpredictable, form the basis of the stigmatisation of this patient population. The connection between mental illness and violence is a complicated problem, and its clarification is in the interest not only of the patients but of therapists and policy-makers as well. In this review we have used studies of foreign authors to find the answer to the question: do severe mental illnesses elevate the risk of committing violent acts and what are the main risk factors for that. We have summarised the results of numerous studies examining birth cohorts, inpatient and outpatient settings and prison populations, and have looked for variables deriving from or independent of the illness that could contribute to the violent behaviour of patients. We have been looking for the most effective ways to prevent people with severe mental illness from becoming violent criminal offenders and the manner to abate the stigmatization of this population. PMID- 19218626 TI - [Gender differences in proneness to depression]. AB - AIMS: Our research has been aimed at identifying the role the risk mechanisms described in Goodman and Gotlib's (1999) model (genetic-biological, interpersonal, social learning related cognitive and stress related factors) play in leading to an increased risk of depression in men and women. METHODS: Genetic biological factors were assessed with the help of certain temperament characteristics and interpersonal factors with parental educational purpose, educational attitudes, educational style and parental treatment. In the case of factors related to social learning, we looked at dysfunctional attitudes and attributional style. As far as the stressors are concerned, we observed the quality of family atmosphere and the number of the positive and negative life events of the preceding six months and their subjective evaluation. 681 students took part in the research (465 females and 216 males). RESULTS: Results of our research have shown that all of the increased risk mechanisms, namely the genetic biological, interpersonal, social learning related cognitive, and stress related factors are connected with the development of a susceptibility to depression, explaining 41.4% of the variance in depression symptoms in women, and 36.5% in men. Harm avoidance, a genetic-biological factor, proved to be the most significant risk mechanism, irrespective of sex. Among the environmental factors irrespective of sex - a stress related factor, the subjective evaluation of negative life experiences, which implies an increased sensitivity to stress, proved to be the strongest risk mechanism. While the above factors played an important role in the development of a susceptibility to depression in both sexes, cognitive and interpersonal risk mechanisms related to the social learning differed in their degree in women and men. In women, the social learning related mechanisms proved to be stronger and higher impact risk factors than in men. The effect of interpersonal factors seemed to be relatively the weakest in the development of an increased risk of depression. LIMITATIONS: The results of our research cannot be extrapolated to Hungarian youth aged 18-23 years due to the limitations of our sample. CONCLUSION: A mental hygienic interpretation of our research findings suggests that in the future more emphasis should be put on the personality development of college and university students, especially on the development of such competences which help them in effectively in their struggle against depressive mood. PMID- 19218625 TI - [Occurrence of anxiety and depression disorders after acute cardiac events during hospital rehabilitation. Application of the Hospital Anxiety and Depression Scale as a screening instrument]. AB - Hospital Anxiety and Depression Scale (HADS) is one of the most frequently used screening instruments for comorbid psychiatric disorders in patients with somatic diseases. Depression and anxiety disorders occurring after acute coronary events predict a worse prognosis and rehabilitation outcome and impair the patient's quality of life. Therefore, diagnosis and treatment of depression and anxiety are important clinical issues. METHODS: Item characteristics, different aspects of reliability and validity of the HADS test parameters have been analysed on the basis of a follow-up study of 747 patients after acute coronary events participating in a hospital rehabilitation program. Short form of the Beck Depression Inventory, Spielberger State Anxiety Inventory, Hamilton Anxiety and Hamilton Depression Questionnaires were used as references. Criterion validity and cut-off points of HADS subscales were determined according to MINI+, a standardised diagnostic interview. RESULTS: The Depression (D) and Anxiety (A) subscales of the Hungarian version of HADS demonstrated high internal consistency (HADS-D: Cronbach =0,81, HADS-A: =0,85). Explorative factor analysis yielded a two-factor (1st: anxiety and 2nd: depression) structure. HADS subscales have shown high correlations with scales measuring similar constructs and validated in Hungarian. Empirical cut-off points of both subscales were determined with ROC analysis (HADS-D: 9 points, HADS-A 8 points). Sensitivity and specificity at these cut-off points fell into the acceptable 0.7 - 0.9 range. CONCLUSION: On the basis of these results, HADS can be used as a reliable and valid instrument for the measurement and identification of anxiety and depression symptoms and disorders in cardiac patients. PMID- 19218627 TI - [Entrapped by the beauty industry: eating and body attitudes of those working in the beauty industry]. AB - INTRODUCTION: Some groups of professionals (models, actresses, ballet dancers, jockeys and athletes) are considered as risk populations for eating disorders and body image disorders. We assume that those working in the beauty industry may also be a high risk group for developing eating disorders, as their work is closely related to beauty and fashion. METHODS: Eating disorders were assessed using the Eating Attitudes Test and the Eating Behaviour Severity Scale, and the tests for assessment of body image included the Human Figure Drawings Test, the Body dissatisfaction subscale of the Eating Disorders Inventory, the Body Attitudes Test, and the Body Investment Scale. RESULTS: Questionnaire data of 543 subjects were analysed. The study sample comprised 128 models from Transylvania (5 males, 123 females) and 167 models from Hungary (10 males, 157 females). These groups were compared with control groups in Transylvania and Hungary, consisting of 148 (25 males, 123 females) subjects in Transylvania and 101 subjects (5 males, 95 females) in Hungary. Weight reducing methods, such as dieting, exercise, the use of appetite suppressants and diuretics, were significantly more prevalent in the group of models. The mean total score and the scores of the Dieting and Bulimia subscales were higher in the group of models in comparison with the control group. The prevalence of clinical and subclinical eating disorders was higher in the group of models. The percentage of subjects scoring above the cut-off score on the Body Dissatisfaction Scale of the Eating Disorder Inventory was higher in the Hungarian control group. The Hungarian control subjects scored higher on the General Body Dissatisfaction Scale of the Body Attitudes Test. Models invest more money and time in their body care. The above results suggest that working in beauty industry may pose an increased risk of developing eating disorders. PMID- 19218628 TI - [Costs of treatment in patients with schizophrenia switched to amisulpride--one year follow-up]. AB - OBJECTIVES: In our present research we have studied the costs associated with switching schizophrenia patients to amisulpride as well as the efficacy of amisulpride treatment. We wanted to explore whether the relatively higher costs of amisulpride can be recovered under the current Hungarian economic and financing conditions. METHODS: From 2002, we analysed clinical improvement with a 6 months follow-up measured by CGI and also compared the costs that were incurred before and after switching in 76 patients suffering from schizophrenia who received amisulpride instead of their previous treatment with typical or atypical antipsychotics. In a second, retrospective phase of the study which lasted for 6 months, we studied the willingness of investigators and patients to continue amisulpride treatment. During this period of treatment both the investigators and the patients were unaware of the fact that the circumstances of treatment would be investigated later; thus, we could determine the number of investigators and patients who decided on the continuation of amisulpride in this phase, and how costs changed later on. In our analysis we followed the cost evaluation methodology introduced earlier by Agnes Rupp. RESULTS: 68 patients were available for the second phase of the study, 65 continued the treatment with amisulpride. Amisulpride has demonstrated cost neutrality in both phases of the study. Higher costs of this medicine have been compensated by an increase in productivity and the resulting cost reduction. Amisulpride treatment was associated with a significant improvement of CGI-measures. CONCLUSIONS: In an open, non-controlled study, modelling a field study in its second phase, amisulpride has been shown to be an effective antipsychotic which is readily accepted by patients and clinicians and which can be prescribed without increasing costs. PMID- 19218629 TI - Exhaled carbon monoxide as a new marker of respiratory diseases in children. AB - Among modern methods included in diagnostic algorithms for various diseases, analyses of expired breath and its condensate acquire increasing importance. Various markers can be determined in the exhaled air, especially volatile gaseous compounds: nitrogen oxide (NO), carbon monoxide (CO), hydrocarbons and 8 isoprostanes. In contrast to NO, CO can serve as a marker of inflammation and oxidation stress. The representation of CO in the exhaled breath (eCO) changes in various diseases of the respiratory and other systems. Among the respiratory diseases in which the use of eCO measurement seems to be perspective and beneficial are bronchial asthma, airways infections, cystic fibrosis, and primary ciliary dyskinesia. The observation of eCO concentrations represents a modern, simple, available, and well reproducible method for the diagnosis of many diseases of respiratory system in children and for the observation of progression, severity of the disease, and response to therapy. PMID- 19218630 TI - Effects of cigarette smoke on the lung and systemic immunity. AB - The influence of tobacco smoke on human health is still an important problem worldwide. Complex inflammatory processes and changes in the immune system are crucial in the pathogenesis of smoking related disorders like chronic obstructive lung disease (COPD), lung cancer, atherosclerosis. The objective of this review is to present the alterations in the immune system in smokers. The main affected system by cigarette smoke (CS) is the respiratory tract. In bronchial epithelium metaplastic and dysplastic changes are accompanied by elevated expression of adhesion molecules and secretion of many cytokines capable of stimulation immune cells influx. In the population of pulmonary macrophages an elevated proportion of cells, changes in expression surface markers with impaired phagocytic and antigen presenting function are observed. Chronic exposure to CS causes increased production of metalloproteinases (MMP) by macrophages and proteolitic enzymes by neutrophils. These enzymes cause destruction of alveolar wall. Increased apoptosis of lung tissue results in augmentation of foreign material which may play a role of autoantigen and which is a target for cytotoxic/suppressor cells. The role of regulatory T (Treg) cells in this process is recently postulated. Smoking cessation is the most effective method of prophylaxis and treatment of diseases related to tobacco smoking. However many immunological changes in smokers are not completely reversible after quitting smoking. PMID- 19218631 TI - Chronic obstructive pulmonary disease: an update on nuclear signaling related to inflammation and anti-inflammatory treatment. AB - Chronic obstructive pulmonary disease (COPD) is one of the most frequent diseases worldwide. Cigarette smoke is considered the main pathological cause of the disorder, although evidence is growing concerning other etiological factors, such as environmental pollution, biomass combustion, infections, genetic predisposition, which may explain why some individuals develop COPD with no history of smoking. Chronic inflammation and remodeling of the small airways characterize the disease at the cellular level, and oxidative stress is considered the main driving force that stands behind COPD inflammation. Recently, chromatin remodeling and epigenetic changes have been found to underlie disease pathology and progression. In this review, the authors gave a short update on the recent hypothesis and findings that may imply novel approach to pharmacotherapy of the disease, focusing on the role of glucocorticosteroids, theophylline, and antioxidants. PMID- 19218632 TI - Therapeutic approach to a child with acute respiratory distress syndrome: a report of two cases. AB - The course of a respiratory disorder in a child may end up in respiratory failure. There are also acute non-respiratory diseases which have a great influence on the respiratory functions and often lead to the acute lung injury and sometimes to the acute respiratory distress syndrome (ARDS). A feature of respiratory function deterioration is changed in the surfactant system. We often see inhibition of its synthesis or damage to its structure. Therapy of children suffering from ARDS should be complex and rapid. Despite many recently published studies explaining the principle of this disorder, the mortality of ARDS is still very high (30-50%). There are several studies documenting successful administration of exogenous surfactant as part of a complex combined therapy of patients with ARDS, which leads to decreased mortality, improved oxygenation, and decreased need for aggressive artificial pulmonary ventilation. The authors of this article present their own experience with administration of exogenous surfactant in therapy of children with ARDS. PMID- 19218633 TI - Systemic treatment by inhalation of macromolecules--principles, problems, and examples. AB - Aerosol inhalation is an established route of medical administration for the treatment of pulmonary diseases. In contrast, aerosol inhalation for treatment of systemic diseases is a novel therapeutic approach. Clinical use of the latter therapy for many years has been limited by the lack of accuracy, efficiency, and reproducibility of the administered doses. Usually, only a small fraction of inhaled drug reached the target region within the lungs. Further problems were the risk of potential allergic reactions in the respiratory tract and a potential variability of drug absorption from the alveoli into the circulation. These problems have been solved in the last years by modern aerosol delivery systems allowing the production of an aerosol with a defined and optimised aerosol particle size combined with an optimized breathing maneuver and optimization of the efficacy of the technology. Furthermore, there were no observations of relevant allergic reactions after inhalation of systemically active drugs in numerous studies. Studies demonstrated that only a small number of morphological factors influence alveolar drug deposition (e.g., exogen allergic alveolitis, active sarcoidosis, active smoking). In consequence, an increasing number of studies investigated the systemic effect of inhaled high molecular weight substances (e.g., insulin, heparin, interleukin-2) and demonstrated that controlled aerosol therapy may serve as a non-invasive alternative for drug application by means of a syringe. Our review briefly summarizes the mechanisms for pulmonary absorption of macromolecules and gives an overview on prior research in the field of inhalant treatment of systemic diseases. PMID- 19218634 TI - Inhaled insulin--does it become reality? AB - After more than 80 years of history the American and European Drug Agencies (FDA and EMEA) approved the first pulmonary delivered version of insulin (Exubera) from Pfizer/Nektar early 2006. However, in October 2007, Pfizer announced it would be taking Exubera off the market, citing that the drug had failed to gain market acceptance. Since 1924 various attempts have been made to get away from injectable insulin. Three alternative delivery methods where always discussed: Delivery to the upper nasal airways or the deep lungs, and through the stomach. From these, the delivery through the deep lungs is the most promising, because the physiological barriers for the uptake are the smallest, the inspired aerosol is deposited on a large area and the absorption into the blood happens through the extremely thin alveolar membrane. However, there is concern about the long term effects of inhaling a growth protein into the lungs. It was assumed that the large surface area over which the insulin is spread out would minimize negative effects. But recent news indicates that, at least in smokers, the bronchial tumour rate under inhaled insulin seems to be increased. These findings, despite the fact that they are not yet statistical significant and in no case found in a non-smoker, give additional arguments to stop marketing this approach. Several companies worked on providing inhalable insulin and the insulin powder inhalation system Exubera was the most advanced technology. Treatment has been approved for adults only and patients with pulmonary diseases (e.g., asthma, emphysema, COPD) and smokers (current smokers and individuals who recently quitted smoking) were excluded from this therapy. Pharmacokinetics and pharmacodynamics of Exubera are similar to those found with short-acting subcutaneous human insulin or insulin analogs. It is thus possible to use Exubera as a substitute for short-acting human insulin or insulin analogs. Typical side effects of inhaled insulin were coughing, shortness of breath, sore throat and dry mouth. Physical exercise increases the transport of inhaled insulin into the circulation and in consequence the likelihood of hypoglycemia. Other problems were the inability to deliver precise insulin doses, because the smallest blister pack available contained the equivalent of 3 U of regular insulin and this dose would make it difficult for many people using insulin to achieve accurate control, which is the real goal of any insulin therapy. For example, someone on 60 U of insulin per day would lower the blood glucose about 90 mg/dl (5 mmol) per 3 U pack, while someone on 30 U a day would drop 180 mg/dl (10 mmol) per pack. Precise control was not possible, especially compared with an insulin pump that can deliver one twentieth of a unit with precision. Another disadvantage was the size of the device. The Exubera inhaler, when closed, was about the size of a 200 ml water glass. It opened to about twice the size for delivery. To our information also other companies (Eli Lilly in cooperation with ALKERMES, Novo Nordisk (AERx, Liquid), Andaris (Powder)) stopped further development and it is unclear whether an inhaled form of insulin will ever be marketed, because of the problems that have occurred. Only Mannkind (Technosphere, Powder) is still working on a Phase III trial. However, our review will briefly summarize the experience regarding inhalant administration of insulin and will describe potential future developments for this type of therapy focussing on the lung. PMID- 19218635 TI - Daily pattern of breathing in healthy young men. AB - Daily changes in the pulmonary function indices were examined in 30 healthy young men. The breathing pattern technique (Lungtest system, MES, Cracow) was used to measure daily changes in basic ventilatory parameters every 3 hours during a 24 hour period. The results revealed significant daily variables in VT (tidal volume), MV (minute ventilation), VT/TI (mean inspiratory flow), and the ratios of MV/P0. 1, (minute ventilation/mouth occlusion pressure) and P0.1/VT/TI. There was lack of significant variations in the other variables. The daily minima in the variables occurred during the usual sleep period at 2:00, 5:00 and also at 8:00 hours. The results of this study demonstrate that healthy adults have small, but significant, daily variations in some of the breathing parameters, although, overall, the breathing pattern remains relatively stable around the clock. PMID- 19218636 TI - The influence of obesity on sleep quality in male sleep apnea patients before and during therapy. AB - Evidence exists that obesity, even in the absence of sleep related respiratory disorders, affects sleep negatively. In this study we examined the influence of obesity on sleep quality of male sleep apnea patients before and after breathing normalization with continuous positive airway pressure (CPAP). We compared the polysomnography from the diagnostic night, second night with CPAP, and a control night (three months later) in 13 non-obese, 13 obese, and 12 severely obese male obstructive sleep apnea patients. In the diagnostic polysomnography, obese and severely obese subjects showed increases in apnea-hypopnea index (AHI) and NREM-1 sleep, and decreases in min SaO(2), REM sleep, and partially slow wave sleep (SWS), when compared with the non-obese group. In the second night under CPAP, normalization of the AHI and a rebound of REM and SWS occurred, which was more pronounced in severely obese than in the non-obese and obese group. The polysomnography recorded three months thereafter revealed no differences in sleep stages between the groups. We conclude that after the long-term CPAP therapy, no effect of obesity on sleep quality is apparent. PMID- 19218637 TI - Use of over-the-counter medications in prevention and treatment of upper respiratory tract infections. AB - Over-the-counter (OTC) medications are a fast developing branch of the pharmaceutical industry, in both Poland and rest of the world. The aim of the work was to evaluate the factors that influence the purchase and use of OTC medications in prevention and treatment of upper respiratory tract infections. The analysis involved 450 randomly chosen subjects, aged 19-80. The study was performed using a questionnaire which included a number of questions related to the characteristics of respondents such as: health, socio-demographic data, and factors influencing the purchase of OTC medications used in respiratory tract infections. The self-medication phenomenon in upper respiratory tract infections is significant in Poland and will continue to increase with support of many demographic and socio-economic factors. Despite certain health risks resulting from the inappropriate use of OTC medications, it should continue to be offered as a part of the pharmaceutical market. PMID- 19218638 TI - Concentration of matrix metalloproteinase-9 in serum of patients with chronic obstructive pulmonary disease and a degree of airway obstruction and disease progression. AB - Chronic obstructive pulmonary disease (COPD) is chronic inflammation leading to irreversible airway obstruction. Previous studies showed increased metalloproteinases (MMP) level, especially MMP-9, as a sign of local inflammation. Up-to-date, only a few studies estimated the MMP-9 serum concentration in COPD with respect to correlation with systemic inflammation. The aim of the present study was to estimate the MMP-9 serum concentration in COPD and to evaluate the correlation between MMP-9 and a degree of airway obstruction in COPD. Twenty three COPD patients and 23 healthy controls were enrolled. In both groups spirometry was performed. MMP-9 concentration in sera taken from both groups was studied using ELISA. We found that COPD patients had increased serum MMP-9 concentration compared with the control group (P=0.0005). In the COPD group, the MMP-9 levels were negatively correlated with FEV1 (P=0.01) and FEV1/FVC (P=0.0002). In conclusion, the results suggest that MMP-9 plays an important role in systemic inflammation in COPD. Higher MMP-9 serum concentration is connected with higher airway obstruction and disease progression. PMID- 19218639 TI - Cough reflex sensitivity is increased in the guinea pig model of allergic rhinitis. AB - Increased cough reflex sensitivity is found in patients with allergic rhinitis and may contribute to cough caused by rhinitis. We have reported that cough to citric acid is enhanced in the guinea pig model of allergic rhinitis. Here we address the hypothesis that the cough reflex sensitivity is increased in this model. The data from our previous studies were analyzed for the cough reflex sensitivity. The allergic inflammation in the nose was induced by repeated intranasal instillations of ovalbumin in the ovalbumin-sensitized guinea pigs. Cough was induced by inhalation of doubling concentrations of citric acid (0.05 1.6 M). Cough threshold was defined as the lowest concentration of citric acid causing two coughs (C2, expressed as geometric mean [95% confidence interval]). We found that the cough threshold was reduced in animals with allergic rhinitis. C2 was 0.5 M [0.36-0.71 M] and 0.15 M [0.1-0.23 M] prior and after repeated intranasal instillations of ovalbumin, respectively, P<0.01, n=36). C2 was not affected in control animals (n=29). We have reported that the selective leukotriene cys-LT1 receptor antagonist montelukast inhibited cough enhancement in this model. We found that this was accompanied by inhibition of the changes in cough reflex sensitivity. C2 was reduced in animals with allergic rhinitis treated orally with vehicle (0.57 M [0.28-1.1] vs. 0.09 M [0.04-0.2 M], P<0.05, n=8), but not in animals treated with montelukast (0.57 M [0.22-1.4 M] vs. 0.52 M [0.17-1.6 M], NS, n=8). We conclude that the cough reflex sensitivity is increased in the guinea pig model of allergic rhinitis. Our results suggest that guinea pig is a suitable model for mechanistic studies of increased cough reflex sensitivity in rhinitis. PMID- 19218640 TI - Superoxide dismutase mimetic modulates hyperoxic augmentation of the diaphragmatic response to poikilocapnic hypoxia in non-vagotomized rats. AB - A period of oxygen breathing enhances the subsequent respiratory responses to episodic hypoxia. Since hyperoxia increases a formation of reactive oxygen species (ROS) in lungs, in the present study we asked a question of whether superoxide anion produced during O(2) breathing would participate in the mechanisms of posthyperoxic enhancement of the response to hypoxia and whether afferent information from the lungs would contribute to this response. To scavenge a superoxide we used Tempol (4-hydroxy-2,2,6,6-tetra-methyl piperidine-N oxyl), a superoxide dismutase mimetic. The respiratory activity of anesthetized, spontaneously breathing rats was assessed from the integrated costal diaphragm EMG. The experiments consisted of 3 min hypoxia (11% O(2)), before and after a 15 min period of breathing with 100% oxygen, with and without Tempol (33 mg/kg) preatreatment. The same protocol was performed in non-vagotomized and vagotomized rats. The results show that a SOD mimetic abolished both hyperoxia-induced slowing of respiration and posthyperoxic respiratory augmentation of the hypoxic response. The abolishment is due likely to a remodeling of the respiratory pattern involving lung vagal reeptors, since in vagotomized animals, the effects of Tempol were marginal. PMID- 19218641 TI - Differences in oxidative status, lung function, and pulmonary surfactant during long-term inhalation of medical oxygen and partially ionized oxygen in guinea pigs. AB - Inhalation of partially ionized oxygen may have less adverse effects on lung functions than medical oxygen. Guinea pigs inhaled air, 100% molecular medical oxygen (O(2)mol), partially negatively (O(2)neg) or positively (O(2)posit) ionized oxygen during 17 and 60 h. After 17 h, dityrosines, markers of oxidative injury, in lung homogenate increased in O(2)neg and decreased in O(2)posit groups vs. controls. After 60 h, dityrosines rose after inhalation of O(2)mol and O(2)neg, but not in the O(2)posit group. Lysine-LPO products increased and lung wet/dry weight ratio decreased in O(2)mol and O(2)neg, and not in O(2)posit group. Relative neutrophil count in BALF was elevated in all oxygen-treated groups with lower numbers in O(2)posit vs.O(2)mol and O(2)neg groups. After 60 h, surfactant activity was better in O(2)posit vs. O(2)mol group. In conclusion, long-term inhalation of partially positively ionized oxygen is associated with less oxidative stress, milder lung inflammatory response, and better surfactant activity than molecular or negatively ionized oxygen. PMID- 19218642 TI - Oxidative stress markers in the blood of persons with different stages of obstructive sleep apnea syndrome. AB - The aim of the study was to determine some biochemical factors reflecting oxidant antioxidant balance in the blood of patients being in different stages of the obstructive sleep apnea syndrome (OSAS). The patients, suspected of OSAS, were submitted to biochemical and somnographic examinations. APolyMesam device was used to establish the severity of apnea episodes. The control group was composed of subjects without OSAS. The following parameters were determined: concentration of plasma total antioxidant status, TAS; activity of erythrocyte Cu, Zn superoxide dysmutase, SOD; plasma lipid peroxidation products, TBARS. The OSAS patients had lower levels of SOD and TAS, and increased level of TBARS. We conclude that oxidative stress markers increase in the blood of OSAS patients to a variable degree, depending on the severity of the syndrome and, therefore, may be suitable for a clinical follow-up of such patients. PMID- 19218643 TI - Hospital staff and smoking habits: do we need modification of smoking behavior in Polish hospitals? AB - The purpose of the work was to analyze the smoking behaviors of the medical staff in hospitals. A typical group of employees in one of the biggest hospitals in Poland was examined thoroughly. The medical examination was carried out by means of standardized investigative instruments--questionnaires processed by the European Network for Smoke-free Hospital. In the studied group, 26% of persons admitted that they smoked, while nearly 12% persons confirmed their addiction. Statistically, it was confirmed that addiction to smoking is conditioned by practice, but the percentage of smoking people was the lowest in the professional group of physicians. However, physicians admitted they smoked in hospitals most often. It is a crucial factor for the psychological preventive treatment as well as for changes in the health behaviors among patients. Also, some factors, which determined smoking, such as a large number of duty hours, were identified. On the basis of the analysis of health behaviors, the authors made an evaluation of the possible solutions that could lead to a reduction of the percentage of smoking employees in the health public institutions. PMID- 19218644 TI - Recognition of gait disturbances in patients with normal pressure hydrocephalus using a computer dynography system. AB - Gait and body balance disturbances are important clinical problems in patients with normal pressure hydrocephalus (NPH). They affect patients' locomotion and lead to a higher risk of falls. The gait pattern may be described as durations of the single and double support and of a stance phase. The aim of the present study was to apply the pattern recognition methods for the evaluation of gait disturbances in patients with NPH before and after neurosurgical treatment (shunt implantation). The results indicate that the parameters measured with a Computer DynoGraphy (CDG) system may effectively differentiate changes of gait in patients with NPH. PMID- 19218645 TI - Prevalence of latent tuberculosis infection in health care workers in Poland assessed by interferon-gamma whole blood and tuberculin skin tests. AB - Health care workers (HCWs) are at risk for developing active tuberculosis (TB). The prevalence of latent tuberculosis infection (LTBI) in this group is unknown in Poland, due largely to the problems associated with the interpretation of the tuberculin skin test (TST) in BCG immunized population. The goal of the present study was to assess the prevalence of LTBI in both clinical and non-clinical 155 HCWs (120 females and 35 males) and to compare the groups at different levels of risk. All participants were interviewed using a questionnaire and underwent interferon-gamma whole blood assay (Quantiferon-Tb-Gold) (QTF) and TST. The questionnaire provided information on possible risk factors for LTBI, including demographic and socioeconomic details, the presence of BCG scars, and the degree of occupational exposure. We found that the prevalence of LTBI among HCWs was, on average, 27.1%. A higher risk of acquiring LTBI was associated with certain work locations (TB lab workers--prevalence 50%, TB ward clinicians--34%, nurses--30%). The prevalence in analytical lab technicians was 20%, in administration staff was 15%. The HCWs with positive QTF test results were older and worked longer than those who had negative results. There was a significant correlation between the level of IFN-gamma and both age (P<0.001) and length of employment (P<0.01). The correlation between the diameter of skin test induration and the magnitude of the INF-gamma response also was significant (P<0.001). We conclude that HCWs are at increased risk of infection, suggesting that appropriate preventive strategies should be undertaken. IFN-gamma test is a useful tool in detecting LTBI cases in a country where BCG vaccination is a national policy. PMID- 19218646 TI - The influence of insulin on calcium ion concentration during transduction of signals into neutrophils. AB - The goal of the study was to evaluate the process of neutrophil activation via Ca(2+)-mediated transduction signal and its modification by insulin. The study was performed with the use of isolated peripheral blood neutrophils obtained from 20 healthy volunteers. Isolated granulocytes were stimulated by fMLP or insulin alone, or by both substances added to the medium in combinations: fMLP + insulin (after 20 min) or insulin + fMLP (after 20 min). To explore the mechanism of intracellular Ca(2+) changes, receptor signal transduction pathway was blocked by tyrosine kinase inhibitors: tyrphostin 25 and genistein. fMLP evoked fast intracellular increase of free Ca(2+) concentration in neutrophils, compared with the resting state (P< 0.001). Insulin did not cause any changes in intracellular Ca(2+) when was added to the previously fMLP stimulated cells. Prestimulation with insulin significantly decreased fMLP-induced intracellular free Ca(2+) concentration compared with fMLP alone (P<0.01). A strong correlation was observed between initial intracellular Ca(2+) concentration after incubation with insulin and the response to fMLP (P<0.0001). The tyrphostin 25 did not influence the Ca(2+) concentration in control granulocytes, but inhibited the fMLP-induced intracellular Ca(2+) increase when added before fMLP (P<0.05). In a Ca(2+)-free medium, a strong relationship between intracellular Ca(2+) and the response to fMLP after incubation with tyrphostin was found (P<0.001) The genistein did not influence the intracellular Ca(2+) in non-stimulated cells. However, it inhibited the fMLP-induced Ca(2+) increase when added before fMLP (P<0.05). The genistein added to the suspension of cells after fMLP stimulation did not influence intracellular Ca(2+) level. A positive correlation was found between the initial intracellular Ca(2+) and the response to fMLP of genistein preincubated cells. This effect was seen in both Ca(2+)-rich, and Ca(2+)-free medium We conclude that insulin is a potent immunomodulator and its signaling pathways are mediated by Ca(2+) concentration changes. The process of intracellular Ca(2+) changes following insulin signaling is, at least partly, tyrosine kinase-related. Derangements in the concentration of intracellular Ca(2+) may represent a link between the mechanisms of insulin resistance in diabetes. PMID- 19218647 TI - Asthma management at primary care level: symptoms and treatment of 3305 patients with asthma diagnosed by a family physician. AB - The goal of this epidemiological study was to assess the symptoms, changes in spirometric parameters, and the type of therapy used in 3305 patients (55.5% women and 44.5% men; mean age 48.3 +/-15.9 years) in whom bronchial asthma was diagnosed by a family physician. The study was carried out with a questionnaire sent to both patients and their physicians. All patients were asked about the frequency and severity of asthma symptoms, smoking habits, and the source of the knowledge on their disease. Physicians gave information concerning the diagnosis and classification of asthma as well as its treatment. In all patients spirometric measurements were performed. The results of the survey indicate that the symptoms of asthma were seen relatively often in the examined cohort of patients, about 20% of them complained of everyday ailments. On the other hand, about 30% of the patients did not report any symptoms. Over 50% of the patients were exposed to tobacco smoke. Inhaled glucocorticosteroids were regularly used by 83% of the patients. We conclude that in the Polish population of asthmatic patients the management of symptoms still falls short of the internationally set guidelines. PMID- 19218648 TI - Role of bronchoalveolar lavage in the initial diagnosis of smoking-related interstitial lung diseases. AB - Some of interstitial lung diseases have a potential relation to tobacco smoking and are known as smoking-related interstitial lung diseases (sr-ILD). The aim of this study was to present the typical features in bronchoalveolar lavage fluid (BALF) of patients with sr-ILD with special account to the morphology of alveolar macrophages. We have found typical features of sr-LD in 18 patients with interstitial lung disease. All patients were smokers. Pulmonary function was moderately impaired. Control group consisted of 12 asymptomatic smokers and 18 healthy nonsmokers. The median total cell count in BALF of patients with desquamative interstitial pneumonia was 64.5 x 10(6) and was much higher than in asymptomatic smokers--9.0 x 10(6), and healthy nonsmokers--3.25 x 10(6). A high proportion (mean 90%) and number (55.5 x 10(6)) of macrophages with the predominance of pigmented cells was found. Mean proportion of CD14 positive macrophages was 6%, and of CD54 positive macrophages - 62%. We concluded that features in BALF typical for sr-ILD are uncommon among routinely evaluated specimens, but differ markedly from other forms of ILD. PMID- 19218649 TI - Relationship between nutritional status and pulmonary function in adult cystic fibrosis patients. AB - Cystic fibrosis (CF) is a multisystem autosomal recessive disorder caused by the mutation of a single gene that encodes for the CF transmembrane regulator protein. Clinically, CF is characterized by chronic pulmonary infection, pancreatic insufficiency, and excessive losses of sweat electrolytes. Along with lung function, nutritional status appears one of the most important prognostic indicators in CF patients. In this study we examined the relationship between nutritional status and pulmonary function in adult CF patients. A group of 39 CF patients (mean age 23.9 +/-3.7 years) was studied. The mean value of body mass index (BMI) was 19.5 +/-2.9kg/m(2) (12.8-24.9kg/m(2)). The patients were grouped according to the presence or absence of malnutrition. Malnutrition was established in 11 patients (28.2%), 5 patients suffered from severe malnutrition. 28 patients (71.8%) had a normal nutritional status, but according to ESPEN guidelines, 9 of those patients were at risk of malnutrition. Statistical analysis revealed a significant difference between malnourished and not malnourished patients with respect to FEV(1)% and FVC%. Moreover, the patients with malnutrition were significantly more frequently colonized by P. aeruginosa and fungi and less so by MSSA. PMID- 19218650 TI - Eosinophilic airway inflammation in chronic obstructive pulmonary disease and asthma. AB - Eosinophilic airway inflammation is regarded as a typical feature of asthma, while in chronic obstructive pulmonary disease (COPD) neutrophils seem predominant inflammatory airway cells. The aim of the present study was to compare the cellular components of airway inflammation in patients with newly diagnosed mild or moderate COPD and asthma. Seventeen patients with COPD (M/F 10/7, aged 57 +/-11 yr) and 22 patients with asthma (M/F 12/10, aged 36 +/-14 yr) were enrolled into the study. None of the patients has been treated with steroids for at least 3 months. All patients underwent clinical examination, laboratory examinations, skin-prick tests, pulmonary function tests, methacholine challenge test, and sputum induction with the total and differential cell count assessments. We found increased number of eosinophils in both study groups. However, there were no significant differences in the cellular composition of induced sputum between the asthma and COPD patients. We conclude that eosinophils are important inflammatory cells not only in asthma, but also in COPD. PMID- 19218651 TI - Comparison of cellular and biochemical markers of airway inflammation in patients with mild-to-moderate asthma and chronic obstructive pulmonary disease: an induced sputum and bronchoalveolar lavage fluid study. AB - Although the clinical pictures of asthma and chronic obstructive pulmonary disease (COPD) may be similar, the pathogenesis differs in many aspects. The aim of the present study was to compare the cellular and biochemical features of airway inflammation in patients with asthma and COPD. The study was conducted in 22 patients with asthma (M/F 12/10, mean age 36 +/-14 years) and 17 patients with COPD (M/F 10/7, mean age 57 +/-11 years). Each patient underwent sputum induction followed by bronchoscopy, and bronchoalveolar lavage. Total and differential cell counts and the concentration of interleukin-8 (IL-8) and myeloperoxidase (MPO) were measured in induced sputum (IS) and BALF. We found no significant differences in the total and differential cell counts in IS between asthma and COPD patients. However, COPD patients showed an increased total macrophage count in BALF compared with asthma patients. The relative eosinophil count in BALF was significantly higher in patients with asthma vs. COPD. The concentration of IL-8 in IS and BALF was significantly higher in patients with COPD vs. asthma patients. The BALF concentration of MPO was significantly higher in patients with COPD compared with asthma patients. We conclude that the comparison of cellular composition and the concentration of inflammatory mediators in IS does not differentiate between asthma and COPD. The evaluation of BALF reveals more differences in the cellular and biochemical features of airways inflammation in patients with asthma and COPD than that of IS. PMID- 19218652 TI - Does the established cause of chronic cough depend on diagnostic approach? AB - The objective of the present study was to evaluate the prevalence of chronic cough causes and to compare the efficacy of two diagnostic approaches used in ambulatory vs. hospitalized setting. Eighty patients with chronic cough, 40 in each group, were enrolled into the study. The etiology of cough was determined on medical history and on either basic (in out-patients) or detailed (in in patients) investigations on most common causes of chronic cough. We diagnosed etiology of cough in all subjects. The most frequent causes of cough were gastroesophageal reflux disease (GERD) and upper airway cough syndrome (UACS). Nonasthmatic eosinophilic bronchitis (NAEB) and multiple cough causes were recognized more often in hospitalized patients (P<0.05). We conclude that the main causes of chronic cough were GERD and UACS. An extensive diagnostic approach allows recognizing NEAB more frequently and reveals the complex nature of chronic cough. PMID- 19218653 TI - Comorbidities as an element of multidimensional prognostic assessment of patients with chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is characterized by chronic airflow limitation, some significant extrapulmonary effects, and important comorbidities. The BODE index, a multidimensional scale, has been proposed to better identify severity of the disease and to predict survival. The goal of the study was to evaluate the prevalence of concomitant diseases in patients with COPD and to assess correlations between comorbidities and the BODE index. Eighty patients with COPD were enrolled. They were at least 40 years old and had more than a 10 year history of smoking. The most frequent comorbidities were: systemic hypertension, edema of lower limbs, movement disorders, varices of lower limbs. Subjects with higher scores in BODE index had significantly greater prevalence of arrhythmias and episodes of pneumonia. Evident correlation was observed between low FEV(1) and episodes of pneumonia. We also found that patients with higher BODE scores had more exacerbations of COPD. Comorbidities are very common in COPD. Results indicate a close connection between the BODE index and some comorbidities, which suggests that these conditions may aggravate the COPD course and increase risk of mortality. PMID- 19218654 TI - Expression of CREB-binding protein and peroxisome proliferator-activated receptor gamma during formoterol or formoterol and corticosteroid therapy of chronic obstructive pulmonary disease. AB - We assessed the effect of therapy on nuclear signaling related to inflammatory processes in sputum cells of patients with chronic obstructive pulmonary disease (COPD). Patients were treated with formoterol (F) or formoterol plus budesonide (F/ICS) b.i.d. for 4 weeks, their sputum cells were isolated and subjected to RNA extraction or lysis, followed by differential centrifugation. Signaling protein levels were assessed by Western blots, their specific mRNAs were quantified using qRTPCR, while 8-isoprostane levels were examined using enzyme immunoassay kit. Cytosolic 8-isoprostane levels and nuclear glucocorticoid receptor expression (protein and mRNA) were not significantly different in both groups, while nuclear cAMP response element binding protein (CREB; protein and mRNA) and peroxisome proliferator-activated receptor gamma (PPARgamma protein and mRNA) were significantly higher in cells from F/ICS-treated patients. CREB-binding protein (CBP; protein and mRNA) levels were significantly lower in F/ICS patients. These changes indicate increased anti-inflammatory signaling in F/ICS-treated patients and seem to be beneficial. PMID- 19218656 TI - Influence of pleural macrophages on proliferative activity and apoptosis regulating proteins of malignant cells. AB - Malignant tumors contain numerous macrophages as a major component of the leukocytic infiltrate. Only few studies have evaluated the interaction between products secreted by macrophages and tumor cells. Our objective was to study soluble factors produced by pleural macrophages. We sampled pleural effusions from patients with cancer and used human tumor cell lines as targets. Pleural macrophages were cultured and the supernatants were used as a conditioned medium for cultures of human cell lines A549, HT29, HCT116, SW620, MCF7, MDA-MB231, JURKAT, and HL60. We investigated apoptosis, proliferative activity, and expression of apoptosis regulating proteins Fas, Bcl2, Caspase-3, and survivin of malignant cells cultured in the conditioned medium. Our findings raise the possibility that macrophages from malignant pleural effusions can act as a factor inhibiting apoptosis of malignant cells. PMID- 19218655 TI - Food allergens and respiratory symptoms. AB - Food allergy may be clinically expressed by a variety of respiratory symptoms, which can be provoked either by IgE- or cellular mediated reactions. Among the diagnostic procedures, newly introduced atopy patch test seems to be important for diagnosis of cellular, delayed immune reactions. We studied the prevalence of positive atopy patch tests with food and inhalant allergens and the correlation between the positivity of atopy patch tests and questionnaire derived atopic and nonatopic espiratory symptoms and diseases in an unselected children population. We found a correlation between the positive patch test result with wheat and cough after physical effort, allergic rhino-conjunctivitis, and bronchitis recidivans. The subjects with positive skin reaction to egg suffered from allergic rhino-conjunctivitis and bronchial asthma. Food and inhalant allergens play an important role in the induction and exacerbation of some respiratory allergic diseases. The positive correlation of positive results of skin tests and history of some respiratory diseases and symptoms also on the population level confirm the importance of these tests in the diagnostic work-up of these allergic diseases. PMID- 19218657 TI - Wavelet analysis of voluntary cough sound in patients with respiratory diseases. AB - Changes in the characteristics of the cough sound may refer to some specific pathological processes and their evolution. In this pilot study we analyzed voluntary cough sound properties in subjects with asthma bronchiale (AB) and chronic obstructive pulmonary disease (COPD) and discriminated them from the control cough sound in healthy subjects. The wavelet transform was used due to a nonstationarity of cough sound recordings. The duration of cough sound was longer during pathological conditions. The longest duration and the highest power of the cough sound were found in COPD. In AB patients, higher frequencies were detected compared with chronic bronchitis and the power of cough sound was shifted to a higher frequency range compared with control coughs. Cough sounds were classified using discriminant analysis with a correct classification rate of about 85-90 %. The method of cough analysis enables an objective quantification of voluntary cough sound with a useful diagnostic and prognostic value. PMID- 19218658 TI - Biological status of adult patients with cystic fibrosis. AB - Cystic fibrosis (CF) is the most common autosomal disorder in the Caucasian population. The main goal of the study was to assess the biological condition of adult patients with CS. Data of 90 CF patients aged 18-31 were considered. The biological condition was determined by the measurement of somatometric traits and the nutritional status. The results show a considerable physical retardation and a poor nutritional status of the studied patients. Nearly 45% of the patients showed symptoms of malnutrition, ranging from slight undernutrition to emaciation. The results, however, show a considerable variability of data among the CF patients compared with the healthy population. A significant relationship between the type of mutation and nutritional status was demonstrated. PMID- 19218659 TI - Pleural fluid adenosine deaminase and interferon gamma as diagnostic tools in tuberculosis pleurisy. AB - Several biological markers have been proposed to improve the efficacy of diagnosing tuberculous pleurisy. The study was undertaken to evaluate the accuracy of pleural fluid adenosine deaminase (ADA) activity and interferon-gamma (IFN-gamma) concentration in differentiating tuberculous pleural effusion (TPE) and nontuberculous pleural effusion (non-TPE). Ninety four patients (50 M and 44 F, mean age 60+/-18, range 18-95 years) with pleural effusion (PE) were studied. TPE was diagnosed in patients with: (i) positive pleural fluid or pleural biopsy culture or (ii) granulomas in the pleural biopsy specimen, after exclusion of other granulomatous diseases. Pleural fluid ADA activity was measured with the colorimetric method of Giusti, while IFN-gamma level was measured with ELISA. TPE was diagnosed in 28 patients. The non-TPE group consisted of 35 patients with malignant PE, 20 patients with parapneumonic effusion/pleural empyema, 5 with pleural transudate, and 6 with miscellaneous PE. The ADA activity and IFN-gamma concentration were significantly higher in TPE than in non-TPE (614.1+/-324.5 vs. 15.1+/-36.0 pg/ml, P<0.0001 and 75.1+/-39.1 vs. 11.0+/-16.6 U/l respectively, P<0.0001). The diagnostic sensitivity and specificity of IFN-gamma measurement (cut-off value of 75.0 pg/ml) were 100% and 98.5% respectively and were similar to those of ADA (100% and 93.9% at the cut-off value of 40.3 U/L). We conclude that pleural fluid ADA activity and IFN-gamma concentration are highly sensitive and specific markers of tuberculous pleurisy. PMID- 19218660 TI - Electrocardiographic changes in patients with spontaneous pneumothorax. AB - The aim of the study was to evaluate the prevalence of electrocardiography (ECG) abnormalities in subjects with spontaneous pneumothorax. Forty consecutive patients (mean age 43.7 +/-19.1 years) with spontaneous pneumothorax participated in the study. There were 22 cases of left-sided and 18 cases of right-sided pneumothorax. The mean relative volume of pneumothorax was 51.4 +/-24.7% according to the Light's index and 53.5 +/-22.9% according to the Rhea method. Heart rate was significantly higher in patients with pneumothorax than after lung re-expansion (91 +/-20 bpm vs. 72 +/-16 bpm; P<0.001). Abnormal left axis deviation was found in 3 patients with left-sided and in 1 with right-sided pneumothorax, while abnormal right axis deviation was found in 2 patients with left-sided pneumothorax. Relevant QRS abnormalities (incomplete RBBB and T-wave inversion) were found in 4 patients (10%). QRS amplitude in V2-V6 leads was significantly decreased in left-sided pneumothorax, while the right-sided pneumothorax was associated with an increase in QRS amplitude in V5-V6. We conclude that ECG in subjects with pneumothorax often reveals significant abnormalities. The most significant abnormalities were seen in patients with massive right-sided pneumothorax. PMID- 19218661 TI - Altered expression of T lymphocyte surface markers in children with chronic autoimmune thyroiditis. AB - CTLA-4 and CD28+ are regulators of T cell activation. The CTLA-4 gene is associated with variety of autoimmune diseases. The aim of the study was to evaluate changes in basic T cell subpopulations, and the expression of CD152+ and CD28+ before and after T cell stimulation in children with autoimmune thyroiditis (AT), as compared with control subjects. Blood samples were obtained from 35 AT children and 25 healthy children. CD markers were evaluated by flow cytometry at baseline, after the culture with phytohemagglutinin and without stimulation. At baseline, CD152+ expression was lower in patients than in controls (P<10(-6)). After stimulation, there were an increase in CD152+ T cells and decreases in CD28+ and CD4+ cells in controls (P<0.01). In AT children, CD152+ T cells remained stable. CD4+CD152+ T cells correlated inversely with antithyroglobulin antibodies. We conclude that alterations in lymphocyte markers are associated with AT. Stimulation leads to differing changes in T-lymphocyte subsets in both examined children populations. PMID- 19218662 TI - Self-injuries in adolescents: social competence, emotional intelligence, and stigmatization. AB - Social competence, emotional intelligence, and strategies of coping with stress were investigated in adolescents who commit acts of self-injury. Furthermore, the extent to which stigma influences changes in psychological functioning of self injuring persons also was examined. The methodology consisted of recognized self reporting psychometric tools. Findings revealed substantial disturbances in all of the above mentioned psychological aspects of functioning of the self-injured. There were significant decreases in the level of emotions directed toward the 'self' and in the components of social competence dealing with intimate relations and assertiveness, as compared with healthy persons. Emotion-oriented strategy of dealing with stress became dominant in the self-injured. A decrease in social competence was appreciably intensified by stigma. Findings, however, failed to reveal changes in emotions directed toward the 'others', which may help explain good functioning in social exposure and everyday life situations of the self injured. PMID- 19218663 TI - Tobacco smoking behavior among high school students in Poland. AB - The aim of the study was to examine Polish students' smoking behavior. A group of 671 students completed a questionnaire about smoking. 53.1% of them tried smoking at least once. The most common age at an attempt to smoke was 18 years. Males tried smoking earlier than females. 23.1% were active smokers during the study. The age of 18 was the most common time when smoking became regular. Males smoked more frequently than females. There were significant differences between the kind of school and the frequency of smoking. Students of smoking parents smoked more frequently than those of non-smoking ones. Most smokers (80%) were aware of detrimental effects of addict and declared a will to quit smoking. We conclude that a substantial percentage of Polish students smoke. Anti-tobacco prevention must be started early, before the age of 18 and continued up to 21. PMID- 19218664 TI - Mask leakage in continuous positive airway pressure and C-Flex. AB - Many patients with obstructive sleep apnea syndrome (OSAS) receiving continuous positive airway pressure (CPAP) complain of leaky masks or too high pressure during expiration. C-Flex is a breathing mode with a constant CPAP pressure during inspiration and a reduced pressure during expiration. We compared the leakage data between CPAP and C-Flex and their influence on patients' compliance. Thirty patients (22 men, 8 women, aged 55.4 +/- 11.7 yr, BMI 32.0 +/- 7.4 kg/m(2)) with polysomnographically diagnosed OSAS got a CPAP or C-Flex therapy in a randomized double-blind and cross-over design. After 6 weeks, an adjustment to the other mode followed. Leakage data were sampled during all polysomnographic examinations. Twelve patients dropped out of the study (7 after C-Flex, 5 after CPAP), 4 of them gave up CPAP therapy completely (2 after CPAP, 2 after C-Flex). The leakage in CPAP mode was 27.5 +/-11.5 l/min and in C-Flex mode 28.0 +/-10 l/min (ns). The average nightly use in CPAP mode was 350.0 +/- 70.2 min and in C Flex mode 347.0 +/- 70.8 min (ns). In the final decision of therapy, 9 patients chose C-Flex and 4 patients CPAP (P=0.001). Five patients had no preference regarding the therapy mode. There is no difference in leakage and compliance between CPAP and C-Flex. But significantly more patients decided for a therapy with the C-Flex mode. There must be other unknown factors that influence the decision for the mode of therapy. PMID- 19218665 TI - Significance of pulmonary vagal afferents for respiratory muscle activity in the cat. AB - The influence of vagal stretch receptor afferents on respiratory motor-output and respiratory changes in esophageal pressure (DeltaP(es)) was studied in anaesthetized cats. Tracheal occlusions and lung inflations were performed during hyperoxic normocapnia, during electrical stimulation of one carotid sinus nerve (CSN) or the intracranial medullary chemosensitivity (MCS), during hypercapnia or the combination of CSN and hypercapnia. Tracheal occlusions during inspiration led to increased and prolonged inspiratory muscle (IM) activity. Moderate hyperinflation in inspiration decreased and shortened inspiratory motor output. Changes in esophageal pressure and in amplitude and discharge duration of IM are largely proportional (0.84>r<0.98) to lung volume above normal endexpiratory volume (FRC). The effects are described as the Hering-Breuer inspiration inhibitory reflex (HB-IIR). Tracheal occlusion or hyperinflation in end inspiratory position not only prolonged expiration but also activated expiratory muscles (EM). The effects linearly (0.86>r<0.98) increased with elevation of lung volume. We refer to these effects as the Hering-Breuer expiration facilitatory reflex (HB-EFR). Severe hyperinflation or rapid inflation of the lungs during inspiration, however, led to an inspiratory facilitation with increased IM activity. During concomitant chemoreflex activation, CSN or MCS stimulation, respiratory hypercapnia, or the combination of both, the extent of the above described responses of IM and EM activity were significantly (0.05>p<0.0002) enlarged. The changes in the discharge period of IM and EM following lung inflation were smaller in the presence of the increased chemical respiratory drive (0.01>p<0.005). The relative changes in EM responses to lung inflations during increased respiratory drive were greater than those of IM. Bilateral vagotomy abolished the respiratory responses to tracheal occlusion and hyperinflation of the lungs. The results of the present investigation show that aside from the well-known inhibition of inspiration, vagal slowly adapting lung stretch receptors facilitate expiration. The sensitivity of the lung reflexes is enhanced with increasing respiratory drive. The HB-inspiration inhibitory reflex limits the depth of lung inflation, whereas the HB-expiration facilitatory reflex promotes an effective lung deflation. Both reflex mechanisms, the inspiratory and expiratory one, are present in eupnoeic breathing, but play an important role during increased chemoreflex drive and obstruction of expiration, e.g., with increased external airway resistance. PMID- 19218666 TI - Endotoxin-induced airway hyperresponsiveness in rabbits: contribution of neuropeptides. AB - Endotoxin (ET) contaminated dusts frequently can be found in the environment. Especially in agriculture, ETs represent one of the main factors in the development of respiratory diseases. In order to investigate the pathomechanisms of ET induced lung injury, the contribution of the NANC-system of vagal C-fibres to increased airway responsiveness (AR) to cholinergic agents was investigated in anesthetized rabbits. In two control groups of 6 anesthetized Wight New Zealand rabbits each, E. coli ET was infused i.v. (0.4microg/kg) (Group 1) or the aerosolized ET was inhaled for 10 min (Group 2). Before and 1 and 3 hours after ET exposure, AR to 0.2 und 2.0% acetylcholine aerosol was measured. The increased AR after ET application was compared with the ET responses in rabbits with neuropeptides depleted by subchronic capsaicin treatment on four consecutive days (Groups 3 and 4). ET-inhalation and ET-infusion both resulted in a significant increase in AR to ACH (P<0.005). The increase in dynamic elastance (E(dyn)), as a measure of airway resistance to 0.2% ACH after ET in both groups was comparable with the response to 2.0% ACH before exposure. In rabbits with capsaicin treatment, basal ACH-responsiveness was similar to the control group. After ET Exposure, increase in AR to ACH was significantly (P<0.05) smaller compared with untreated rabbits. In conclusion, depletion of neuropeptides by capsaicin resulted in a significantly decreased ET-induced airway hyperresponsiveness (AHR) in rabbits, indicating the contribution of the NANC-system and their neuromodulators to ET-induced AHR. The results confirm the important role of the NANC-system environmental and occupational agents induced-AHR in rabbits. PMID- 19218667 TI - Distinction between voluntary cough sound and speech in volunteers by spectral and complexity analysis. AB - Objective monitoring of cough sound for extended period is an important step toward a better understanding of this symptom. Because ambulatory cough monitoring systems are not commercially available, we prepared own monitoring system, which is able to distinguish between voluntary cough sound and speech in healthy volunteers. 20-min sound records were obtained using portable digital voice recorder. Characteristics of the sound events have been calculated in time and frequency domains and by a nonlinear analysis. Based on selected parameters, classification tree was constructed for the classification of cough and non-cough sound events. We validated the usefulness of our algorithm developed against manual counts of cough obtained by a trained observer. The median sensitivity value was 100% (the interquartile range was 98-100) and the median specificity was 95% (the interquartile range was 90-97). In conclusion, we developed an algorithm to distinguish between voluntary cough sound and speech with a high degree of accuracy. PMID- 19218668 TI - Influence of physical activity on psychosomatic health in obese women. AB - It is unclear to what extent the known psychosomatic benefits of exercise hold true for the obese. In the present study, we investigated the hypothesis that the psychosomatic health and components of general intelligence, such as the capacity for logical-deductive tasks, would be better in regularly exercising than non exercising obese women. We addressed the issue in a self-reported survey study, comprising two groups of middle-aged obese women (age 30-50 years, BMI >30 kg/m(2)) of 25 persons each. The criterion for the group division was regular exercise, minimum twice a week, for at least 2 months. The following psychometric tools were used: Physical Fitness and Exercise Scale, Patient Health Questionnaire-9 for depression, Life Satisfaction Scale, General Health Inventory 28, Raven's Matrices Test for intelligence, and a test for selfcontentment with one's body figure shape. The exercising obese women scored significantly better in Life Satisfaction Scale (17.1 +/- 1.2 vs.12.0 +/- 0.9), had a lower level of depression (8.1 +/- 0.6 vs. 13.4 +/- 0.7), and a better assessment of the health status (24.6 +/- 1.6 vs. 36.4 +/- 2.2) (reversed score) compared with non exercising ones (P<0.05). The exercising obese women also appreciably better assessed their bodily looks. Interestingly, if depression was present in exercising women, it had more detrimental health effects than in physically inactive ones. The study failed to substantiate appreciable changes in general intelligence between active and non-active obese women. In conclusion, physical activity is of benefit for the psychosomatic health in obese women, which should be considered in behavioral counseling. PMID- 19218669 TI - Comparison of the effects of low-dose vs. high-dose aminophylline on lung function in experimental meconium aspiration syndrome. AB - Due to missing information on appropriate dosing of aminophylline in meconium aspiration syndrome (MAS), this study compared effects of high-dose and low-dose aminophylline on lung function of animals with MAS. Meconium-instilled rabbits were treated by low-dose (LD, 1.0 mg/kg), or high-dose (HD, 2.0 mg/kg) aminophylline at 0.5 and 2.5 h after meconium instillation, or were left untreated. Within 5 h of oxygen ventilation, HD-aminophylline improved gas exchange, reduced pulmonary shunts and ventilatory pressures, and decreased edema formation and lung neutrophils. LD-aminophylline enhanced lung function to a lower extent than HD-aminophylline, and failed to reduce lung edema and the number of lung neutrophils. Both treatments decreased lung peroxidation, with a stronger effect of HD-aminophylline on lipid oxidation and of LD-aminophylline on protein oxidation. Tracheal reactivity to histamine decreased after HD aminophylline, while lung tissue reactivity was more reduced after LD aminophylline. Although LD-aminophylline showed some anti-inflammatory potential, HD-aminophylline improved most of the parameters more effectively. PMID- 19218670 TI - Combination of budesonide and aminophylline diminished acute lung injury in animal model of meconium aspiration syndrome. AB - Combination of low-dose budesonide and low-dose aminophylline may improve lung function in reduced adverse effects compared with high-dose monotherapy. Adult rabbits intratracheally received 4 ml/kg of saline or meconium (25 mg/ml). Meconium-injured rabbits were treated at 0.5 and 2.5 h after meconium instillation by intravenous aminophylline (1.0 mg/kg), by intratracheal budesonide (0.125 mg/kg) followed by intravenous aminophylline (1.0 mg/kg), or were untreated. Although aminophylline improved some respiratory parameters, budesonide+aminophylline more effectively reduced intrapulmonary shunts and improved gas exchange, without significant cardiovascular effects. Combined treatment reduced lung edema and number of lung neutrophils to a higher extent than aminophylline alone. Both treatments reduced lung peroxidation and in vitro airway reactivity to histamine, with a better effect after aminophylline alone. Combination of budesonide and aminophylline enhanced respiratory parameters more effectively, having fewer side effects than aminophylline alone. However, no additive effect of budesonide was observed on lung peroxidation and in vitro airway reactivity. PMID- 19218671 TI - Influence of selective inhibitors of phosphodiesterase 3 and 4 on cough and airway reactivity. AB - As the administration of many antitussive drugs is often associated with adverse effects, new alternatives are evaluated in experimental and clinical conditions. The aim of this study was to assess the influence of selective inhibitors of PDE3 (cilostazol) and PDE4 (citalopram) on cough and airway reactivity. The number of cough efforts, specific airway resistance, in vitro airway reactivity, and differential blood cells count were measured in healthy and in ovalbumin sensitized guinea pigs before and after administration of cilostazol or citalopram (1 mg/kg). Cilostazol significantly suppressed citric acid induced cough only in healthy guinea pigs, whereas citalopram in both healthy and ovalbumin-sensitized animals. Both PDE inhibitors decreased in vivo and in vitro airway reactivity to histamine and the count of monocytes and neutrophils, confirming their anti-inflammatory potential. Administration of selective PDE3 and PDE4 inhibitors may influence cough and airway reactivity in the model of ovalbumin-sensitized guinea pigs. PMID- 19218672 TI - Asthma, allergy, and respiratory symptoms in centenarians living in Poland. AB - According to National Census, there were 1541 people over the age of 100 years (centenarians) in Poland, in 2002, including 1215 females and 326 males. The aim of the present study was to assess the prevalence of asthma, allergy, and respiratory symptoms in centenarians included in the Polish Centenarians Program, POLSTU 2001, conducted between 2001 and 2004. The study group consisted of 301 subjects including 258 females and 43 males. Research data were gathered in a questionnaire designed exclusively for the study, completed by the interviewer during meetings with the subjects and their families. According to the medical history reported by the subjects, 10 persons (3.3%) suffered from asthma and 41 (13.6%) from allergy. There were no subjects with childhood onset asthma. Allergy to food and medicinal products was the most prevalent. One in four centenarians reported dyspnea and one in eight complained of cough. When analyzed in relation to gender, cough was more prevalent in males, which might have been related to cigarette smoking. Respiratory disorders are frequent in elderly populations, but symptoms may be underreported, especially in the situation of coexisting medical problems. Moreover, it might be difficult to perform full diagnostic procedures in the very elderly due to disability, cognitive impairment, and technical problems. Thus, medical care for the aged should be based on thorough medical evaluation supported by the medical history and reliable information on physician diagnosed diseases. PMID- 19218673 TI - Severe respiratory distress caused by central airway obstruction treated with self-expandable metallic stents. AB - We investigate retrospectively the efficacy of self-expandable metallic stents (SEMS) for severe respiratory distress (SRD) in patients with central airway obstruction (CAO). Thirty three patients with CAO were treated with SEMS using fiberoptic bronchoscopy method. We found an intraluminal obstruction present in 7, extraluminal compression in 10, and combined stenosis in 16 cases. Tumor infiltration occupied more than 90% of the endoluminal diameter in 21, 70% in 9, and 50% in 3 cases. Obstruction was caused by primary cancer of lung in 23, thyroid in 5, and esophagus in 5 patients. Up to 3 stents per patient were placed. Double stenting (esophagus and trachea) was required in five patients. All patients exhibited symptomatic and arterial blood gas improvement. The mean follow-up was 65 (5 to 752) days. SEMS are useful for the treatment of SRD caused by CAO. The overall effect is related to the degree of tumor progression itself. PMID- 19218674 TI - Upregulation of Th1 cytokine profile in bronchoalveolar lavage fluid of patients with hypersensitivity pneumonitis. AB - The aim was to find out how the IL-12 and IL-18 levels in the bronchoalveolar lavage fluid (BALF) correspond to the inflammatory activity of the hypersensitivity pneumonitis (HP). We studied 12 patients with HP and 13 normal subjects. IL-12 and IL-18 levels were measured using ELISA kits. We found a significantly higher plasma angiotensin-converting enzyme (ACE) concentration (55 vs. 34 U/L, P=0.0016), lymphocyte percentage (57 vs. 14%, P<0.001), CD8+ cells (32 vs. 17%, P<0.001) and a lower CD4/CD8 ratio (1.2 vs. 2.0, P<0.0001). The IL 12 and IL-18 levels in BALF were significantly higher in HP patients than in healthy subjects (3.9 vs. 3.2 pg/ml, P=0.003 and 14.2 vs. 6.15 pg/ml, P% 0.0001, respectively). We found a strong positive correlation between IL-12 and the percentage of lymphocytes (r=0.68, P=0.015) and a negative one between IL-12 and the percentage of macrophages in BALF (r=-0.64, P=0.024). We conclude that upregulation of the Th1 cell cytokine profile may play a significant role in the pathogenesis of HP. PMID- 19218675 TI - Increased levels of interleukin-12 and interleukin-18 in bronchoalveolar lavage fluid of patients with pulmonary sarcoidosis. AB - We studied prospectively 43 patients with sarcoidosis and 13 normal subjects. IL 12 and IL-18 levels were measured using ELISA kits. Spirometry and body plethysmography were performed using an Elite DL Medgraphics body box. The sarcoidosis group was characterized by significantly higher median range of the plasma angiotensin-converting enzyme (ACE) concentration (72 vs. 34 U/l, P<0.0001), lymphocyte (34 vs. 14%, P<0.0001), CD4+ cells percentages (59 vs. 36%), and CD4/CD8 ratio (4.2 vs. 1.99, P<0.0001). The BALF IL-12 and IL-18 levels were significantly higher in sarcoidosis patients than in healthy subjects (4.1 pg/ml vs. 3.2 pg/ml; P<0.001 and 11.1 pg/ml vs. 6.15 pg/ml, P<0.0001, respectively). A negative correlation between BALF IL-12 and ACE plasma levels (r=-0.33, P<0.05) within the sarcoidosis group was found. Our data suggest a potential role of IL-12 and IL-18 in a local immunologic response in pulmonary sarcoidosis. PMID- 19218676 TI - The origins of bioethics: advances in resuscitations techniques. AB - During the last years there has been an increasing interest in meta-bioethical issues. This turn in the research focus is regarded as a sign of the maturation of bioethics as a distinct area of an academic inquiry. The role of historic philosophical reflection is often emphasized. It should be noted that there is a rather common agreement that the future of bioethics lies in the critical reflection on its past, in particular, on the very origins of this discipline. Sharing Caplan's opinion, advances in medicine technologies, especially the introduction of respirators and artificial heart machines, is considered as one of the main issues that started bioethics. Using methods of historical as well as meta-ethical research, this article aims at describing the role of advances in resuscitation techniques in the emergence of bioethics and at exploring how bioethical reflection has been shaped by technological developments. A brief historical analysis permits to say that there is a close bond between the emergence of bioethics and the introduction of sophisticated resuscitation technologies into medical practice. The meta-ethical reflection reveals that advances in resuscitation techniques not only initiated bioethics in the second half of the 20(th) century but influenced its evolution by (i) posing a question of justice in health care, (ii) altering commonly accepted ontological notions of human corporeality, and (iii) reconsidering the very purpose of medicine. PMID- 19218677 TI - Changes of airway obstruction parameters in healthy children caused by mother's smoking during pregnancy. AB - The present study was aimed at the assessment of the impact of mother smoking during pregnancy on changes of phase angle (phi) and T(me)/T(E) index in healthy children. A hundred and twenty seven children, divided according to age (<6 months and >6 months of age) and mother smoking anamnesis were investigated by noncalibrated respiratory inductive plethysmography in the supine position. We found statistically significant changes of phi (p<0.05) and T(me)/T(E) in healthy children of non-smoking mothers against a group of smoking mother's children of up to 6 months of age. These differences were not confirmed in children older than 6 months. Moreover, in the smoking mother group, we found statistically significant changes (P<0.05) of phi and T(me)/T(E) in children of up to 6 months of age in comparison with children older than 6 month. The results revealed a negative impact of mother smoking during pregnancy represented by changes in airway obstruction parameters, which appeared especially in the group of youngest children. PMID- 19218678 TI - Right ventricular function in patients with severe interstitial lung disease: a Tissue Doppler imaging study. AB - Tissue Doppler imaging (TDI) can be useful in the evaluation of right ventricular (RV) regional dysfunction due to pulmonary diseases. The aim of the study was to assess RV dysfunction in patients with severe interstitial lung diseases (ILD), using both TDI and conventional transthoracic echocardiography (TTE). The study group consisted of 40 consecutive patients with end-stage ILD referred for lung transplantation (LT). Eighteen of them, mean age 47 +/-12 yr, who fulfilled the ATS/ERS criteria for LT (mean forced vital capacity (FVC) 1.4 +/-0.8 l, mean diffusing capacity for carbon dioxide (DLCO) <50% of predicted) constituted the active group, and 22 age-matched patients (mean age 49 +/-12 yr), who did not fulfill these criteria, were placed in the waiting group for LT. We found that among the TDI parameters, the longitudinal peak systolic strain rate (SR) measured at the RV outflow tract, was significantly more negative (-1.1 +/-0.3 vs. -3.2 +/-1.2 s(-1), P=0.03) and the time to peak velocity (TpVEL) in the medial and apical septal segments were shorter (129.9 +/-47.9 vs. 159.2 +/-38.1 ms, P=0.018 and 126.8 +/-44.3 vs. 154.6 +/-40.9 ms, P=0.019) in the patients from the active than those from the waiting group. TTE showed that patients from the active group had significantly lower values of tricuspid annulus plane systolic excursion (TAPSE) (14.3 +/-3.3 vs. 20.6 +/-6.2 mm, P=0.0003) and pulmonary artery acceleration time (PVAT) (70.3 +/-23.3 vs. 96.9 +/-12.4 ms, P=0.0001)compared with those from the waiting group. No other differences were found for other parameters of RV function between the examined groups. In conclusion, severity of lung disease in patients with ILD influences RV systolic dysfunction, which is reflected in both TDI and conventional echocardiographic parameters. PMID- 19218679 TI - Driving performance in patients with chronic obstructive pulmonary disease. AB - Patients with chronic obstructive pulmonary disease (COPD) present with impairments of their cognitive performance. It is still unknown whether cognitive deficits influence driving abilities in patients with COPD. The present study investigates driving performance in patients with COPD and healthy controls. Driving simulation was performed in 17 patients with COPD and 10 healthy controls. Patients with COPD demonstrated significantly worse results in terms of accident frequency in the simulated driving situation. No correlations existed between the severity of disease, assessed from the polysomnographical findings (e.g., lung function, blood gas analysis, sleep disturbance, nocturnal ventilation, and oxygen saturation), and driving performance. We conclude that impairments of driving performance in patients with COPD cannot be predicted on the basis of the severity of the disease. The impairment of driving performance in the simulated driving situation in COPD patients may have crucial consequences for driving licensing in these patients. PMID- 19218680 TI - Prevalence and correlates of allergic diseases among children. AB - The purpose of this paper was to determine the correlation between socioeconomic status, lifestyle, and the prevalence of allergic diseases in children. It was also checked whether there were any differences in the biological development rate between the children with allergies and their peers. The study is based on the data obtained from a cross-sectional surveys of 301 children aged 4-9. The number of children with allergies found as declared by their parents was 58 (19.3%). The prevalence of allergic children in the villages was significantly lower than in towns (12.8% vs. 26.2%). The more frequent occurrence of allergies in children also depended on the level of their parents' education, very good economic conditions, artificial feeding during the first months of their lives and attending nurseries. The children with diagnosed allergies did not differ from their peers by height, but had a higher body weight. PMID- 19218681 TI - Cough reflex sensitivity testing in in seasonal allergic rhinitis patients and healthy volunteers. AB - Dysfunction of upper and lower airways frequently coexists, and they appear to share key elements of pathogenesis. The interrelationship between upper and lower airway manifestations of allergy remains still incompletely investigated. Little is known about the critical factors that determine airway afferent nerve endings reactivity (cough sensitivity) in patients with allergic rhinitis. Subclinical inflammatory changes within the lower airways and/or chronic upper airway cough syndrome (previously referred to as postnasal drip syndrome) are probably responsible for this effect. The aim of this study was to evaluate capsaicin cough sensitivity in pollen sensitive patients with seasonal allergic rhinitis without treatment out of a grass pollen season time using the European Respiratory Society (ERS) guidelines on the assessment of cough. Cough sensitivity was defined as the lowest capsaicin concentration which evoked two (C2) or five (C5) coughs. Capsaicin aerosol in doubling concentrations (from 0.49 to 1000 micromol/l) was inhaled by a single breath method (KoKo DigiDoser; nSpire heath Inc, Louisville, CO), modified by the addition of an inspiratory flow regulator valve (RIFR; nSpire heath Inc, Louisville, CO). The seasonal rhinitis subjects (5M, 7F; mean age 23 yr) had not been complaining primarily about coughing. Their pulmonary function was within normal range. Concentrations of capsaicin causing two (C2) and five coughs (C5) were reported. Volunteers' (5M, 7F, mean age 23 yr) cough sensitivity (geometric mean and 95 % CI) for C2 was 16.5 (4.1-67.0) micromol/l vs. allergic rhinitis patients' C2 3.5 (1.9 - 6.4) (P= 0.018). Volunteers' C5 was 132.4 (41.3 - 424.5) micromol/l vs. allergic rhinitis patients' C5 13.1 (6.0 - 28.6) micromol/l (P= 0.013). We conclude that airway afferent nerve endings reactivity in pollen sensitive subjects suffering from seasonal allergic rhinitis was significantly increased out of pollen season compared with healthy volunteers. PMID- 19218682 TI - Correlation between the stage of cystic fibrosis and the level of morphological changes in adult patients. AB - The aim of this study was to evaluate the correlation between the stage of cystic fibrosis and the intensity of accompanying morphological changes--including transmission electron microscopy--within bronchial mucosa. The stage of the disease was assessed on the basis of clinical status and radiological and endoscopic examination. We focused on morphological changes in epithelial cells, the presence of metaplasia and/or dysplasia, the type of inflammatory infiltrate, and the presence of epithelial ulcerations, thickening of epithelial basement membrane and collagenization of lamina propria. We found two clinically different patients groups. The first one was in a poor clinical condition, advanced inflammatory fiberoptic bronchoscopy and radiological changes, multiple exacerbations, and with chronic inflammation and only focal appearance of ciliated epithelium. Moreover, squamous cell metaplasia and dysplasia was diagnosed in 3 and 4 cases, respectively. The other group, in a better clinical condition, had normal BMI and small changes on chest Xray. In this group the diagnosis of cystic fibrosis was made at later age. Two patients from this group displayed features of acute phase; ciliated epithelium was covering the whole sample. After statistical analysis, we found a correlation between the clinical course and the morphological changes in bronchial mucosa. Bronchial ulcerations, squamous cell metaplasia and dysplasia were found in the group with the more severe clinical course. PMID- 19218683 TI - Depression and memory: a comparative study of young and old women. AB - The study compared the prevalence of depression, memory dysfunction, and the influence of depression on memory in two contrasting age-groups of women: young- the mean age of approximately 23 years (35 subjects) and old--the mean age of approximately 70 years (44 subjects). The study population samples were subdivided into depressive and nondepressive groups, according to the presence and absence of depressive symptoms, respectively, as assessed with the Center for Epidemiologic Studies Depression Scale (CES-D). Memory function was assessed with a modified Visual Letter Recall Test, University of Washington, Seattle, USA. The results show that the prevalence of depression was greater in the group of young compared with the old women; 63% vs. 41% of subjects, respectively. The intensity of depression was moderate in both age-groups. The accompanying memory dysfunction was significantly greater in the group of old than young women; 43% vs. 18% of missed letters on recall, respectively (P<0.05). The results failed to lend support for a causative link between depression and memory dysfunction. Memory deficit did not intensify with increasing depression score and also was present in the women without depression in both age-groups. In conclusion, older age in women not necessarily leads to more frequent appearance of depression, and a decline in memory function with advancing age also has to do with factors other than depression. PMID- 19218684 TI - Sense of social interaction in the elderly. AB - The intuit is that the need for social interaction increases in the elderly. The issue, however, has not been thoroughly investigated. In the present study we posed the following main questions: (i) does the need for social interaction indeed increase in older age? and (ii) is there a defined personality trait that would interact with the level of social interaction? The study was one of a self reported group survey. A total of 50 women were enrolled, who were divided into two even contrasting age-groups: young, 20-30 years, and senior, 60-80 years old. The main exclusion criterion was depression, which was screened for before subject enrollment. The results demonstrate that the need for interpersonal meetings was similar in the young and old women; the mean score amounted to 15.4 +/-0.5 and 15.6 +/-0.6 points, respectively. The most frequent reason for the meetings in both young and old women was to socialize; 80 and 68% of women, respectively. The rarest, in turn, reason was to get help in everyday life matters; 8% young and 12% old women. Thus, we failed to show appreciable differences in the need, character, and goals of social interaction between the two age-groups of women. Social interaction had most often a free and selfless character. We also found that extraversion, independent of age, facilitated social interaction. In conclusion, although to the question asked of whether 'the older the person, the more he/she misses the interaction with others' 76-88% of the surveyed women in both age-groups answered affirmatively, the findings of the present study failed to confirm that. PMID- 19218685 TI - Influence of microinjections of D,L-homocysteic acid into the Botzinger complex area on the cough reflex in the cat. AB - Microinjections of D,L-homocysteic acid (DLH) were used to test the hypothesis that neuronal activation within the Botzinger complex area can modify the spatiotemporal characteristics of the cough reflex in 17 spontaneously breathing pentobarbitone anesthetized cats. DLH (50 mM, 1.25-1.75 nmol, 9 cats) reduced the number (P<0.01) of coughs and expiratory amplitude of abdominal electromyographic activity (P<0.01), and also esophageal pressure (P<0.001) during mechanically induced tracheobronchial cough. The duration of cough abdominal activity was shortened by 48% (P<0.05). DLH microinjections also temporarily reduced the respiratory rate (P<0.01) and increased the mean arterial blood pressure (P<0.001), baseline of esophageal pressure (P<0.01), and end tidal CO(2) concentrations (P<0.01). Lower doses of DLH (0.27-0.35 nmol, 7 cats) or vehicle (25-35 nl, 8 cats) induced few alterations in cardiorespiratory or cough characteristics. The results support predominantly inhibitory effects of neurons in the region of the Botzinger complex on cough abdominal activity and cough number. PMID- 19218686 TI - Medullary raphe midline is involved in production of expulsive expirations in anesthetized rabbits. AB - Effects of kainic acid lesions in the medullary raphe midline on reflex expirations induced mechanically from the trachea were examined. Spontaneously breathing rabbits were anesthetized by ketamine and xylazine i.m., followed by pentobarbitone i.v. Excitatory neurotoxin kainic acid (2 mg/ml in artificial CSF, total volume of 55-100 nl) was pressure microinjected into the medullary midline, rostral to the obex (2 microinjections at 2 different depths). The lesion (mostly affected the obscurus and magnus raphe nuclei) reduced the number of reflex expirations by 80% and expiratory amplitudes of esophageal pressure, abdominal EMG moving averages, and abdominal EMG powers by 71%, 62%, and 57%, respectively (in all cases P<0.05). The duration of abdominal activity in post-lesion responses was not altered. Control microinjections of artificial CSF had no effect on the reflex responses. We conclude that in rabbits, the medullary raphe nuclei participate in the control of expiratory expulsions originating from the trachea. PMID- 19218687 TI - Frequency of distribution of inflammatory cytokines IL-1, IL-6 and TNF-alpha gene polymorphism in patients with obstructive sleep apnea. AB - Obesity is one of the most commonly identified factors for the obstructive sleep apnea syndrome (OSAS). Adipose tissue is the source of many cytokines, among them there are IL-6, IL-1, and TNF-alpha. The level of inflammatory cytokines increases in people with OSAS and obesity. The aim of this study was to evaluate the distribution of genotypes in inflammatory cytokine genes in people with obesity-related OSAS. The examined group consisted of 102 person with obesity related-OSAS and 77 normal weight person without OSAS. Genotyping of DNA sequence variation was carried out by restriction enzyme (IL-1: Taq I, IL-6: Lwe I, TNF alpha: Nco I) analysis of PCR amplified DNA. The study revealed a significant correlation between polymorphism located in the promoter region of inflammatory cytokine genes and obesity-related OSAS. PMID- 19218688 TI - Sleep disorderd breathing and recurrence of cerebrovascular events, case fatality, and functional outcome in patients with ischemic stroke or transient ischemic attack. AB - Ninety one patients with stroke or transient ischemic attack (TIA) were screened for sleep-disordered breathing (SDB). Case fatality, rate of recurrence of cerebrovascular events, and functional outcome were analyzed during a 2-year follow-up. The patients were stratified into groups: without (AH < or =5) and with SDB (AHI >5). SDB was present in 61 (67.7%) patients with stroke or TIA. The rate of recurrence of TIA or stroke in patients with SDB was significantly higher (12 patients, OR=1.52, P<0.05) as compared with patients without SDB (3 patients) within two years of observation. Case-fatality rates were not significantly different (4 patients with SDB and 2 patients without SDB). Our data show that SDB significantly increases the incidence of recurrent cerebrovascular events in patients with TIA or stroke in a two-year follow-up. SDB in patient with stroke or TIA did not influence functional outcome of stroke during the long-term observation. PMID- 19218689 TI - Cardiac chemoreflex sensitivity in critically ill patients. AB - Chemoreflexes are important mechanisms for regulating ventilatory and cardiovascular function. The aim of this study was to determine the meaning of autonomic dysfunction for the pathophysiology and outcome in critical ill patients. For the determination of the chemoreflex sensitivity (ChRS), the ratio of the RR interval shift and the shift of oxygen partial pressure during a 5-min inhalation of oxygen with a nose mask was formed. Pathological chemoreflex sensitivity was predefined as a ChRS below 3.0 ms/mmHg. Out of the 27 critical ill patients included into the study, 17 had a sepsis and 10 a cardiogenic shock. In these patients, chemoreflex sensitivity was significantly reduced compared with a control group (sepsis: 2.1 +/- 1.68, cardiogenic shock: 0.4 +/- 0.27, controls: 5.0 +/- 2.8 ms/mmHg; P<0.05 vs. sepsis or cardiogenic shock). There was a significant negative correlation (r=-0.6; P<0.01) between the chemoreflex sensitivity and the severity of illness described by the SOFA-score. We conclude that cardiac reflex mechanisms are changed toward increased sympathetic activity reflected by reduced chemoreflex sensitivity in critical ill patients. Moreover, there is a close negative correlation between the ChRS and the SOFA-score. PMID- 19218690 TI - Safety of reagents for infection testing: results of the market surveillance by the Federal Institute for Drugs and Medicinal Devices until end 2006. AB - The European Directive 98/79/EC on in vitro diagnostic medical devices (IVD) stipulates the marketing and post market surveillance of IVD in the European Economic Area. In cases of issues and field corrective actions, the manufacturers have to inform the responsible Competent Authorities (CA). In Germany, the Federal Institute for Drugs and Medical Devices (BfArM) is the responsible CA for most IVD, with a small subset of IVD for immune hematological and infection testing as well as tissue typing as specified in Annex II of the Directive, being within the responsibility of the Paul-Ehrlich-Institute (PEI). In this study, all issues regarding reagents for infection testing, but not laboratory analyzers, reported to the BfArM between begin 1999 and end of 2006 were analyzed in respect to the source of report, the underlying product defects, and the performed corrective actions. Within the observation period a total of 888 reports on IVD were received of which 90 related to the IVD for infection testing included in our study. Reports were predominantly received from manufacturers (55) and Competent Authorities (29). Affected products were most frequently those for serological analysis (42) and culturing techniques (36), whereas molecular biological tests played only a minor role (12). Investigations of the manufacturers were able to identify the underlying root causes of product failures in 68 cases (75.6%). In 16 cases (17.8%) the root cause remained unclear and in 6 cases (6.6%) a product failure was excluded or a user error was the underlying cause. Most frequently product failures were caused by material defects (25), production errors (11), microbial contamination (6), and labelling errors (5). Manufacturers issued corrective measures in 73 cases (81.1%). Based on the underlying root causes of product failures, these were predominantly (multiple entries) customer information (71), recall (58), modifications in production or quality management (50), modifications of the raw materials (17), and modifications of the instructions for use (12). The results and experience obtained since 1999 suggest that the system for post marketing surveillance of IVD is an established tool to ensure product safety even though the current system can be further optimised. PMID- 19218692 TI - Respiratory responses to acute intermittent hypoxia and hypercapnia in awake rats. AB - This article deals with the recognition of early changes in the breathing pattern, in response to acute intermittent stimuli in awake rats. Two different types of stimuli were given: 9% hypoxia in N(2) and 10% hypercapnia in O(2). Animals were exposed to 3 consecutive cycles consisting of 3-min stimulus period separated by 8-min normoxic recovery intervals. Features of the breathing pattern, such as respiratory frequency, tidal volume, minute ventilation, inspiration and expiration times, peak inspiratory and expiratory flows, were measured by whole body plethysmography. The data were analyzed with the use of pattern recognition methods. We conclude that the overall respiratory changes were rather slight. However, computerized analysis using a k-nearest neighbor decision rule (k-NN) allowed for a good recognition of the respiratory responses to the stimuli. The misclassification rate (E(r)) varied from 5 to 10%. After feature selection, E(r) decreased below 1%. The k-NN classifier differentiated correctly also the type of intermittent stimulus. Our experimental results demonstrate usefulness of pattern recognition algorithms in studying respiratory effects in biological models. PMID- 19218691 TI - Induced sputum in patients with interstitial lung disease: a non-invasive surrogate for certain parameters in bronchoalveolar lavage fluid. AB - Induced sputum is a useful non-invasive method for the assessment of airway and parenchymal lung diseases. This study aimed to compare induced sputum and bronchoalveolar lavage fluid (BALF) cellular composition and T-lymphocyte subpopulations in patients with interstitial lung disease. We evaluated 33 patients: 15 with sarcoidosis, 11 with hypersensitivity pneumonitis, and 7 with idiopathic pulmonary fibrosis. The percentage of macrophages was significantly lower in induced sputum than in BALF in sarcoidosis (P=0.005), and the percentage of neutrophils was higher in induced sputum than in BALF in sarcoidosis (P=0.001) and hypersensitivity pneumonitis (P=0.006). A significant correlation was found between the BALF and induced sputum CD4+, CD8+ subsets and the CD4+/CD8+ ratio in both the whole patient group (r(s)=0.80, r(s)=0.88, r(s)=0.88, P<0.001, respectively) and in the 3 subgroups. A strong correlation of the T-lymphocyte subsets in induced sputum and BALF in patients with interstitial lung disease shows that induced sputum may be a non-invasive surrogate for certain parameters in BALF in these patients. PMID- 19218693 TI - Obstructive sleep apnea in heart failure patients: evidence for persistent conduction disturbances or sinus node dysfunction. AB - Bradycardia is a common finding in patients with obstructive sleep apnea and might be pronounced in heart failure patients. The aim of the present study was to determine the relationship between nocturnal hypoxemia, apnea-hypopnea index, and electrophysiological parameters of sinus node and atrioventricular conduction properties. Electrophysiological studies were performed in 12 patients with heart failure. Polygraphic studies were done in all of the patients. Patients with an AHI >10/h were classified as sleep apnea patients. Mild sleep apnea was diagnosed in 50% of the patients (AHI 17.8 +/- 4.4 vs. 5.1 +/- 3.6/h). There were no differences with respect to the resting heart rate, PQ interval, or QRS duration between the two groups. Sinus node recovery time was normal in all of the patients (993 +/-291 vs. 1099 +/-62 ms, P=0.45). There was no abnormal atrioventricular conduction. Nevertheless, sleep apnea patients showed decreased atrioventricular conduction time (AH) intervals (134 +/- 42 vs. 102 +/- 25 ms, P=0.1) and infranodal conduction time (HV) intervals (59 +/- 9 vs. 43 +/- 7 ms, P=0.01). We conclude that mild sleep apnea was not associated with abnormal findings in sinus node function or AV conduction properties in patients with heart failure. Decreased AH/HV intervals might be a consequence of apnea associated sympathetic activation. PMID- 19218694 TI - Evaluation of fluorescence-based methods for total vs. amplifiable DNA quantification in plasma of lung cancer patients. AB - In the last decade numerous reports demonstrated that free-circulating DNA in plasma/serum samples might be a promising biomarker in a number of pathologies, including cancer. Thus, choosing the reliable and efficient method of plasma DNA quantification would be an essential step prior to any clinical evaluation of cell-free DNA measurement in cancer patients. The aim of present study was to compare two highly-sensitive DNA quantification methods in regard to their applicability and effectiveness in monitoring the cell-free DNA level in the blood of patients with resectable non-small cell lung cancer. Plasma samples collected from 10 patients before any treatment, after neoadjuvant therapy and subsequent surgery, were used for DNA quantification by direct fluorescent PicoGreen staining and by real-time qPCR in SYBR Green and TaqMan probe approach using beta-actin gene as the amplifying target. The PicoGreen method demonstrated a high level of correlation with both the SYBR Green (r=0.87, P<0.0001) and TaqMan probe approach (r=0.94, P<0.0001). The total DNA content, determined by PicoGreen, proved to be several-fold higher than the amplifiable DNA amount measured by real-time qPCR. Consequently, intercalating fluorochromes, like PicoGreen, might serve as a rapid, accurate, and inexpensive alternative to real time qPCR for routine dsDNA quantification and multicenter standardization. PMID- 19218695 TI - Recurrence of arterio-venous malformations with life-threatening complications in a pregnant woman with hereditary teleangiectasia. AB - Hereditary hemorrhagic teleangiectasia (HHT) is an autosomal dominant vascular disorder with incidence of 1 in 2300, characterized by teleangiectasia, arterio venous malformations, and aneurysms. In this article we presented a case of a 43 year-old woman, diagnosed with HHT and treated with a lobectomy at the age of 5 and with transcatheter coil closure of pulmonary feeding artery at the age of 30, who developed a recurrence of arterio-venous fistulas in the lungs and the brain during pregnancy. The case was complicated by ischemic stroke with hemiparesis. The patient went into premature labor at 35 weeks and the child was delivered by cesarean section. The patient developed severe pulmonary insufficiency with hemothorax post partum, which required transcatheter embolotherapy. During a seven year follow-up, the patient developed progressing intrapulmonary shunt deterioration and hypertrophic pulmonary osteoarthropathy. The case described shows that significant morbidity and mortality may arise during pregnancy from this otherwise relatively benign condition. PMID- 19218696 TI - Treatment of Churg-Strauss syndrome with an inhaled corticosteroid after oral steroids discontinuation due to side effects. AB - Churg-Strauss syndrome is characterized by a history of asthma, peripheral blood and tissue eosinophilia, with a predilection to the lungs. In this article we present a case of a 35 years old patient admitted to the hospital as a case of severe asthma apparently complicated by bilateral pneumonia. After a transient improvement on antibiotics, the respiratory symptoms recurred one month later in an exacerbated form. The diagnosis of Churg-Strauss syndrome was established on a clinical basis and a long-term therapy with oral corticosteroids and cyclophosphamide was introduced. The treatment must have been withdrawn after a year, because of a whole range of adverse effects, even though there was a severe recurrence of symptoms during the therapy. A regimen of the inhaled corticosteroid ciclesonide was then started, which entirely prevented the recurrence of respiratory symptoms over a two year follow-up period. PMID- 19218697 TI - Upper respiratory tract diseases in self-evaluation of health status of Polish students based on the SF-36 questionnaire. AB - People's health beliefs have remained under the influence of various factors. Our health is determined by physical and social environment, health behaviors, and lifestyle. The evaluation of frequency of upper respiratory tract diseases among Polish students was the aim of the study. Survey was conducted among 476 students of medicine and dentistry in the Pomeranian Medical University of Szczecin and pedagogy students of the Zielona Gora University, Poland. SF 36 questionnaire with few additional questions concerning health and disease was used in the study. Students' participation in the research was voluntary and anonymous. More than 80% of medical students treat their upper respiratory infections by themselves. From all surveyed subjects only 48 (9.4%) of the students did not suffer from any illness during an academic year. Part of the students (39.7%) spends their leisure time in a passive way. Pedagogy students more often than medical ones spend their leisure time actively. Male medical students showed much lower values comparing to female medical students. Pedagogy students showed reversed situation. There was a high variability of general health components observed among female, but not male students. Frequent respiratory tract infections, lack of systematic physical activity, and low amount of sleep cause lower level of self evaluation in regard to health status among medical students. PMID- 19218698 TI - Cardio-respiratory interaction and autonomic dysfunction in obesity. AB - We aimed to test whether the evaluation of the cardio-respiratory interaction using the analysis of heart rate and blood pressure variabilities and respiratory maneuvers can reveal cardiovagal dysfunction in obese adolescents 12-18 years old. The spectral power in high frequency band of the heart rate variability (HRV) reflecting respiratory sinus arrhythmia was used as an index of the cardiac vagal control, and the spectral power in high frequency band of the blood pressure variability (BPV) as an indicator of mechanical effects of respiration. The deep breathing test and Valsalva maneuver were applied. The obese group had a reduction in spectral power in high frequency band of the HRV. Differences in high frequency band spectral power of the BPV between the obese and control groups were not found. The finding of lower respiratory sinus arrhythmia, indicating a cardiovagal dysfunction in obese adolescents, can provide important diagnostic information about early subclinical autonomic dysfunction in obesity. PMID- 19218699 TI - Role of gender and pubertal stage on cough sensitivity in childhood and adolescence. AB - Studies in healthy adult volunteers and patients with chronic cough have shown that women have increased cough sensitivity to inhaled tussigenic stimuli, including capsaicin. The explanation for this phenomenon remains unknown, although the influence of pubertal changes (dimensional, hormonal) may play a role. In the present study we set out to examine the effect of the pubertal status on cough reflex sensitivity in a population of male and female children and adolescents. The study consisted of cough reflex sensitivity (CRS) testing, spirometry, and a children-completed pubertal status questionnaire. The inclusion criteria were no current symptoms of respiratory disease, no respiratory infection in the preceding 2 weeks, no allergic disease, and no other disease that could modulate CRS. All children were non-smokers. For assessing the CRS we used a compressed air-driven nebuliser controlled by a dosimeter with an inspiratory flow regulator valve. Each subject inhaled up to 12 capsaicin aerosol concentrations (0.61-1250 micromol/l) during 400 ms at 1 min intervals. CRS was defined as the lowest capsaicin concentration that evoked minimally 2 coughs. 225 children (median age 13 yr, 103 girls/122 boys) were divided according to the pubertal status (prepubertal, early pubertal, and late pubertal) and gender. We found that CRS [geometric mean (95%CI)] was similar in prepubertal and early pubertal girls and boys. However, CRS was significantly higher in late pubertal girls (n=56) than in boys (n=26) [53.57 (35.62-80.64) micromol/l vs. 119.7 (70.74 208.55) micromol/l; respectively; P=0.017]. We conclude that puberty and gender have a significant influence on the cough threshold to capsaicin. It seems, however, hard to determine what factors are responsible for the observed gender differences in cough reactivity adolescents. PMID- 19218700 TI - Inhibition of salivary amylase activity by cigarette smoke aldehydes. AB - Cigarette smoke (CS) is a leading known cause of cancer and cardiovascular diseases worldwide. The mechanisms by which CS produces its damaging effects seem to be multifactorial. Among others, CS toxicity is due also to several compounds like alpha,beta-unsaturated aldehydes (acrolein, crotonaldehyde) and saturated aldehydes (acetaldehyde). Aldehydes could interact with thiol compounds of salivary proteins, leading to structural and functional alterations of these molecules. Prior in vitro studies have shown that there is a significant decrease in several known enzymatic activities following exposure to CS. Additionally, it was found that glutathione (GSH) has protective effect against the damaging role of CS to salivary enzymes, emphasizing the role of thiol groups in the mechanism of inactivation of these enzymes. In this study, salivary amylase activity showed a significant inhibition following exposure to CS, and to external addition of purified aldehydes known to be present in CS, due probably to the interaction between aldehydes and -SH groups of the enzyme. Our results indicate that although saturated aldehydes are the chief aldehydes present in CS, a significant decrease in amylase activity was due to unsaturated aldehydes, reacting, probably, through their double bond with the thiol group of proteins by the Michael addition reaction. PMID- 19218701 TI - Bond disorders of a patient with asthma: a case study. AB - The issue of treatment and providing preventive activities in bronchial asthma is still not fully investigated. Besides the medical aspect, psychiatrists and psychologists point to psychosomatic aspects, since psycho-social factors can evoke intensification of asthma symptoms, and also become the consequences of that disease. Previous psychosomatic investigations show that chronic illness makes the patients reveal extended dependence on others. Patients may also tend to perceive other people as being responsible for their health problems. Such an attitude is usually caused by the disease process and the feeling of threat related to disease symptoms. In the present article I discuss the case of a patient with diagnosed bronchial asthma who was in the course of psychodynamic therapy. The aim of therapy was to help the patient in an interpersonal sphere and, when a therapeutic alliance is already established, to support individualization and separation in his previously disordered process of development, and finally to support the process of recovery. The case presented below is one of many others, when patients with chronic diseases, including bronchial asthma, apply for therapy. The adult patients usually complain about discomfort in social situations, sometimes about difficulties with keeping work that they have just got, or, like in this case, about bond disorders which reveal as intrusiveness and adhesiveness. PMID- 19218702 TI - Interferon gamma production in the course of Mycobacterium tuberculosis infection. AB - It is not clear why some individuals with unknown predisposition develop tuberculosis, while others remain healthy in spite of heavy exposure. Interferon gamma (IFNgamma) is considered to be the key cytokine responsible for resistance to M. tuberculosis infection, as confirmed by increased susceptibility to mycobacterial infections in rare inherited defects in IL-12-IFNgamma axis. The aim of this study was to assess the IFNgamma production by peripheral blood lymphocytes from immunocompetent tuberculosis (TB) patients. The study group included 51 TB patients. In all cases, TB was confirmed by culture. Twenty healthy TB contacts were considered as control group. Commercially available ELISA-based assays were used to measure IFNgamma in the supernatant of whole blood cell cultures after stimulation with PWM (Phytolacca Americana), PHA (phytohemagglutynin), and PPD (purified protein derivative). No difference in IFNgamma secretion between the patients and control group was found when blood cells were stimulated by PWM or PHA. PPD-induced IFNgamma formation was higher in TB patients than in controls. The secretion of IFNgamma after non-specific stimulation varied in different clinical and radiological presentation of tuberculosis and it was lower in most advanced and extensive forms of the disease. It is unclear whether the difference in formation and release of IFNgamma is a primary or secondary phenomenon in the course of the disease. PMID- 19218703 TI - The impact of the sleep apnea syndrome on oxidant-antioxidant balance in the blood of overweight and obese patients. AB - The aim of the study was to assess the markers of oxidant-antioxidant status in excess body mass index (BMI) persons with and without the obstructive sleep apnea syndrome (OSAS). In overweight (BMI-1: 25.0-29.9 kg/m(2)) control and OSAS and obese (BMI-2: 30.0-34.9 kg/m(2)) control and OSAS subjects with no acute or chronic disorder the following markers were determined: concentration of plasma total antioxidant status, TAS, (Randox); activity of erythrocyte Cu, Zn superoxide dysmutase, SOD, (Randox); plasma concentration of thiobarbituric acid reacting substances, TBARS, (Yagi method). BMI-1-OSA presented decreased SOD, compared with the BMI-1-C group (P=0.006). BMI-2-OSA showed both SOD (P=0.002) and TAS (P=0.047) decreased, and elevated TBARS (P=0.03) compared with the BMI-2 C group. There was a positive correlation between TAS & SOD and a negative one between TAS & TBARS in the BMI-1-C group. In BMI-2-C, a negative correlation between TAS & TBARS was observed. We conclude that OSAS decreases the blood antioxidant status in high BMI persons and may change the relationship between oxidative stress markers. PMID- 19218704 TI - Modulatory effect of sera from patients with various types of pulmonary fibrosis on mononuclear cell induced angiogenesis in relation to pulmonary function. AB - Angiogenesis plays an important role in the pathogenesis of idiopathic pulmonary fibrosis. Pulmonary fibrosis occurs also in many diseases, such as other types of interstitial pneumonias or drug-induced pulmonary fibrosis. The aim of the study was to examine the effect of sera from patients with various types of pulmonary fibrosis on angiogenesis induced by human mononuclear cells (MNC) in relation to lung functions. The study population consisted of 32 patients with idiopathic pulmonary fibrosis (IPF), 11 patients with drug-induced pulmonary fibrosis (DIPF), 6 with cryptogenic organizing pneumonia (COP), and 20 healthy volunteers. An animal model of leukocyte-induced angiogenesis assay was used as an angiogenic test. Spirometry, whole-body plethysmography, static lung compliance (Cst), and diffusing capacity of the lung for CO (DL(CO)) were performed in all patients. Sera from IPF and COP patients significantly stimulated angiogenic activity of MNC, compared with sera from healthy donors and from DIPF patients (P<0.001). However, sera from healthy donors and DIPF significantly stimulated angiogenic activity of MNC compared with the control group with PBS (P<0.001). In all groups, a decrease in the mean value of Cst and DL(CO) was observed, but no significant correlation between VC, FEV(1), DL(CO), Cst, and angiogenic activity of sera from examined patients was found. Sera obtained from patients with pulmonary fibrosis constitute a source of mediators modulating angiogenesis, but the pattern of reaction is different in various diseases. The strongest reaction is observed in IPF and the weakest one in DIPF. The angiogenic activity of sera did not correlate with the pulmonary function of patients with pulmonary fibrosis. PMID- 19218705 TI - Angiogenic activity of sera from silicosis and pulmonary Langerhans cell histiocytosis patients in relation to lung function tests. AB - Angiogenesis has been implicated in the pathogenesis of interstitial lung diseases. A correlation between serum angiogenic cytokines level of patients with idiopathic pulmonary fibrosis and radiographic manifestations or functional pulmonary changes has been described, but the role of angiogenesis in the pathogenesis of other interstitial lung diseases such as silicosis and pulmonary Langerhans cell histiocytosis remains unclear. The aim of the study was to examine the effect of sera from silicosis and pulmonary Langerhans cell histiocytosis patients on angiogenesis induced by human mononuclear cells (MNC) in relation to pulmonary function. The study population consisted of 12 patients with silicosis, 12 patients with pulmonary Langerhans cell histiocytosis (PLH), and 14 healthy volunteers. Spirometry, whole-body plethysmography, static lung compliance (Cst), and diffusing capacity of the lung for CO (DL(CO)) were performed in all patients. As an angiogenic test, leukocyte induced angiogenesis assay according to Sidky and Auerbach was used. Sera from PLH patients exerted a significant inhibitory effect on angiogenesis (P<0.001). Sera from silicosis patients significantly (P<0.001) stimulated angiogenesis compared with sera from healthy donors. However, sera from healthy donors significantly stimulated the angiogenic activity of MNC compared with the control with PBS. The mean value of DL(CO) was significantly lower in the group of patients with PLH compared with patients with silicosis (P<0.05). A significant correlation between angiogenesis index and DL(CO) was observed (P<0.05). No significant correlation between the angiogenesis index and other functional parameters was found. Sera from interstitial lung diseases patients and healthy donors constitute a source of mediators modulating angiogenesis. Sera from silicosis patients stimulate neovascularization but sera from PLH patients exert an inhibitory effect on angiogenesis. A correlation between serum angiogenic activity and DL(CO) was found. PMID- 19218706 TI - Angiogenic activity of sera from patients with systemic autoimmune diseases in relation to clinical, radiological, and functional pulmonary status. AB - Systemic autoimmune diseases, such as vasculitis and collagen diseases, are characterized by chronic inflammation. Mutual interrelationship between angiogenesis and chronic inflammation has already been demonstrated. The aim of the study was to examine the effect of sera from patients with systemic autoimmune diseases on angiogenesis induced by human mononuclear cells. The study population consisted of 43 patients with a systemic autoimmune disease associated with pulmonary manifestations, divided into three groups: 14 with Wegener's granulomatosis (WG), 13 with systemic sclerosis (SS), and 16 with collagen vascular diseases (CVD) such as rheumatoid arthritis, systemic lupus erythematosus, and dermatomyositis. The control group consisted of 15 healthy volunteers. Clinical status was evaluated using a questionnaire. Standard chest radiographs were performed in all patients. Pulmonary function tests were performed according to the ERS standards. An animal model of a leukocyte-induced angiogenesis assay was used as an angiogenic test. Sera from WG and CVD patients significantly stimulated angiogenesis compared with healthy subjects (P<0.001). On the other hand, sera from healthy donors exerted a proangiogenic effect compared with PBS. In contrast, sera from SS patients significantly (P<0.001) inhibited angiogenesis compared with sera from healthy subjects and PBS. Proangiogenic effect of sera from systemic diseases patients depended on radiological changes. No significant correlation between a degree of dyspnea or functional pulmonary tests and the number of new vessels or angiogenesis index was found. Sera from patients with systemic autoimmune diseases and healthy people constitute the source of mediators modulating angiogenesis. These modulatory effects differ depending on the disease entity. PMID- 19218707 TI - Spirometric parameters in malnourished girls with anorexia nervosa. AB - Repercussions of obesity on the lung function have been widely studied. The effect of serious malnutrition is less well known. The aim of study was to determine spirometric parameters in 102 malnourished girls with anorexia nervosa. Among these patients, only 71 aged 12-18 years (mean 15.6), mean BMI 15.8 kg/m(2), met the ATS/ERS forced expiratory maneuver criteria for spirometry. The most frequently observed abnormalities were: decreased IC seen in 33 (46%) girls and decreased PEF in 45 (63%) patients. Maximum voluntary ventilation was within the normal range in all but 2 subjects. Diminished values of FEV(1), FVC, FEV(1)/FVC, MEF(50) were observed in 10 (14%), 13 (18%), 3 (4%), and 3 (4%) patients, respectively. We found strong positive correlations between weight and absolute values of the examined parameters. We assume that spirometric abnormalities in anorexia are probably a result of respiratory muscle weakness and body mass loss. PMID- 19218708 TI - Induced sputum metalloproteinases and their inhibitors in relation to exhaled nitrogen oxide and sputum nitric oxides and other inflammatory cytokines in patients with chronic obstructive pulmonary disease. AB - The aim of this study was to assess sputum levels of the metalloproteinases MMP 1, MMP-2, MMP-3, TIMP-1, and TIMP-2, as well as MMPs/TIMPs ratios in relation to exhaled NO (eNO) and sputum NOs (nitrates and nitrites) and IL8 obtained from chronic obstructive pulmonery disease (COPD) patients, healthy non-smokers, and healthy smokers. We found higher levels of TIMP-1 (118.9 ng/ml) and TIMP-2 (3.75 ng/ml) in COPD patients than in healthy smokers (17.7 ng/ml, P<0.03; 0.51 ng/ml, P>0.05, respectively) and healthy non-smokers (84.6 ng/ml, P>0.05; 1.61 ng/ml, P>0.05, respectively). We also observed significant positive correlations between concentrations of NOs and MMP-1, MMP-2, MMP-3, and TIMP-2 (r=0.37, P<0.02; r=0.60, P<0.0001; r=0.56, P<0.0004 and r=0.47 P<0.004, respectively) in COPD patients. IL8, MMP-2, MMP-3, and TIMP-2 levels in induced sputum were negatively correlated with airway obstruction, i.e., FEV(1)/FVC (r=-0.61, P<0.00009; r= 0.41, P<0.01; r=-0.38, P<0.02; r=-0.49, P<0.002). Our study points to a potentially pathogenic role of stromelysin-1 (MMP-3) in COPD. PMID- 19218709 TI - Influence of mechanical hippotherapy on skin temperature responses in lower limbs in children with cerebral palsy. AB - Therapeutic horse riding is often employed for disabled children. The aim of the present paper was to determine the influence of exercise in a mechanical saddle, imitating horse's walk, on the skin temperature responses in lower limbs in children with cerebral palsy. Sixteen children, aged 14-16, were enrolled into the study. Skin surface temperature was assessed with thermography, using an infrared thermovision camera, AGEMA 550, before and directly after 20 +/-5 min of exercise. The findings demonstrate that mechanical hippotherapy provides an exercise stimulus that is capable of inducing a visible change in skin temperature of paralyzed limbs. The change, however, is one of a decrease in skin temperature, which points to acute vasoconstrictive effects of exercise and to decreased skin blood flow. The results, therefore, do not support the use of mechanical hippotherapy in children suffering from cerebral palsy, with a hope to stimulate blood circulation to spastically altered limb muscles and thereby to improve physical disability, at least in a short-term exercise paradigm. PMID- 19218710 TI - Chronic crusting, nasal carriage of Staphylococcus aureus and relapse rate in pulmonary Wegener's granulomatosis. AB - Wegener's granulomatosis is a systemic disease characterized by necrotizing granulomatous inflammation of the upper and lower respiratory tract and necrotizing crescentic glomerulonephritis. Nasal carriage of S. aureus is considered a risk factor for S. aureus infections. The aim was to examine possible risk factors for relapse including refractory nasal carriage of Staphylococcus aureus in patients with Wegener's granulomatosis. Swab cultures from anterior nares for S. aureus were taken in consecutive patients (n=28), with limited (n=15) and systemic forms (n=13) of biopsy-proven Wegener's granulomatosis. The occurrence of infection and relapses were identified according to defined criteria. Seventeen of the 28 patients (60%: 95% Cl, 41-76%) were found to be chronic nasal carriers of S. aureus (> or =80% of nasal cultures positive for S. aureus). A hazard regression analysis identified chronic nasal carriage of S. aureus as independent risk factor for relapse (HR-4.56; Cl 2.45 7.65) in patients with limited Wegener's granulomatosis. Chronic nasal carriage of S. aureus characterized patients with Wegener's granulomatosis, who are more prone to relapses. PMID- 19218711 TI - Association between clinical activity and high-resolution tomography findings in pulmonary Wegener's granulomacytosis. AB - Wegener's granulomatosis is characterized histologically by necrotizing granulomatous angitis that most commonly involves the upper, lower respiratory tract, and kidneys, but may affect any organ system. The aim of the study was to assess the usefulness of high-resolution computed tomography (HRCT) for evaluating pulmonary disease activity in Wegener's granulomatosis patients. Pulmonary disease activity at the time of examination was scored in 66 patients with Wegener's granulomatosis according to clinical, radiological, and bronchoscopic findings: activity group (n=43, Group 1), past activity group (n=14, Group 2). Of 66 staging examinations, 57 (86%) revealed abnormal CT scans: masses or nodules (30 patients in Group 1 and 6 patients in Group 2, parenchymal opacifications (15 in Group 1 and 1 in Group 2), pleural irregularity (3 in Group 1 and 10 in Group 2). We conclude that HRCT may be a useful supplement to clinical scoring of disease activity in Wegener's granulomatosis with pulmonary involvement. PMID- 19218713 TI - Correlation between Helicobacter pylori infection and pulmonary Wegener's granulomacytosis activity. AB - Wegener's granulomatosis is a clinicopathologic entity of unknown origin characterized histologically by necrotizing granulomatous angiitis that affects any organ system. The disease most commonly involves the upper and lower respiratory tract and kidneys. Wegener's granulomatosis is a disease which requires a long-term use of steroids and NSAIDs. Because of that patients frequently develop gastroduodenal mucosal lesions and concurrent Helicobacter pylori infection. The aim of the study was to assess the impact of H. pylori infection on clinical features in patients with Wegener's granulomatosis treated with non-steroidal anti-inflammatory drugs, steroidal drugs, and cyclophosphamide. Thirty six patients with systemic Wegener's granulomatosis were tested for the presence of H. pylori infection and 25 of them turned up H. pylori positive. The severity of Wegener's granulomatosis disease, prevalence of gastroduodenal lesions, and the type and duration treatment seem to depend upon H. pylori infection. PMID- 19218712 TI - Procalcitonin as an indicator of systemic response to infection in active pulmonary Wegener's granulomacytosis. AB - Differential diagnosis of infection during active immune disease, such as Wegener's granulomatosis (Wegener's granulomatosis), is a major clinical challenge. Laboratory measures, erythrocyte sedimentation rate or C-reactive protein, can be elevated in infections that supervene, or coinciding with, in active Wegener's granulomatosis, and thus are nonspecific. The aim of the study was to compare the serum levels of procalcitonin (PCT) in patients with active and inactive disease. Twenty two sera were tested from 10 patients with active, generalized, and biopsyproven Wegener's granulomatosis, with pulmonary involvement, and 12 patients with nonactive one. PCT levels were measured using an immunoluminometric assay. The PCT level was markedly elevated (1.2-3.6 ng/ml) in 9 of the 10 sera from active and 2 of the 12 sera from nonactive Wegener's granulomatosis. PCT levels were in the normal range (0.28-0.56 ng/ml) in the remaining patients with nonactive Wegener's granulomatosis. We conclude that serum procalcitonin levels may be a potentially useful marker in the diagnosis of bacterial infection supervening in active Wegener's granulomatosis. PMID- 19218714 TI - Quest for "innovative strategies" in primary PCI: clot busters or devices (protection vs suction) or both? A story in search of an end. PMID- 19218715 TI - An overview of modern embolic protection in percutaneous coronary intervention. AB - Coronary embolism due to atherosclerotic debris is a rather common cause of post procedural complications. While evidence has shown that both arteriolar vasodilators and platelet glycoprotein inhibitors have proven ineffective against post- and peri-procedural embolism,5 mechanical interventional devices have been shown to improve (lower) 30-day MACE rates. These interventions include distal filtration, distal, and proximal occlusion balloons. The distal occlusion balloon was the first approach to embolic protection. The intervention involves placement of a low pressure (<2 atm) balloon distal to the lesion of interest. Antegrade flow is temporarily interrupted while the lesion is treated. Mounted on conventional 0.014-inch guidewire shafts, distal filtration systems follow a similar intervention method to distal occlusion. In this proceeding, a delivery/recovery sheath catheter deploys an expandable filter device approximating the lumen, which is later removed following PTCA or stent placement in retroversion. The variety of existing, rather novel filter designs typically feature a wire mounted umbrella-type filter consisting of laser-drilled micropores design varied, averaging approximately 100 microm. The primary benefit derived of distal filtration includes the trivial uninterruption of antegrade flow. Unlike distal occlusion, proximal devices allow for vessel protection before lesion crossing, a great advantage in cases involving thrombosis, vulnerable plaque, or primary unstable angina. Proximal occlusion follows a nearly identical implementation as distal occlusion. While substantial research is still needed, interventionalists are advised to always use embolic protection devices in SVG interventions. PMID- 19218716 TI - In vitro cardiogenesis can be initiated in human CD34+ cells. AB - BACKGROUND: The extensive damage that occurs in the cardiac tissue after myocardial infarct is the major concern in post infarct management. It is very well known that adult stem cells mobilized by administration of G-CSF result in homing of stem cells into the damaged myocardium. This is because of the fact that stem cells have the ability to proliferate and capacity to generate into multiple cell lineages. METHOD: A healthy donor was selected as per the guidelines given by the institutional ethical committee and Helsinki declaration. The donor was given G-CSF 5 microg/kg/day and stem cells were harvested from the peripheral blood using Fresenius ASTec204 cell separator. The PBSC were then evaluated by immunohistochemical staining using anti-human CD34 monoclonal antibodies. The cells were then cultured in DMEM with 10% FCS for 17 weeks and in vitro cardiogenesis was initiated by adding 4 microM/l 5'Azacytidine. RESULTS: In vitro cardiogenesis was initiated in pure CD34+ cells with 5' Azacytidine. The cells showed spontaneous beating after 24 hours of treatment and after 5 weeks, the cells connected with the adjoining cells by a myotube. In these cells, expression of myosin light chain (MLC2v) gene and GATA-4 transcription factor validated the development of cardiomyocytes. CONCLUSION: It is observed that the transplantation of autologous stem cells/fetal cardiomyocytes in the heart scar tissue developed due to infarct, limited the scar expansion, and prevented post infarct heart failures. Homing process due to the transplantation of autologous stem cells is time consuming; therefore, transplantation of cardiomyocytes developed from autologous stem cells could be the future method of correcting the infracted myocardium. PMID- 19218717 TI - Efficacy of PercuSurge in acute myocardial infarction patients undergoing revascularization with a special emphasis on procedural time. AB - AIM: To evaluate the efficacy of the PercuSurge Guardwire(R) Plus Temporary Occlusion and Aspiration System, the actual procedural time involved and long term follow-up in acute MI patients undergoing primary/rescue percutaneous coronary intervention (PCI). METHODS & RESULTS: It was a single centred, prospective study in 67 prospective AMI patients undergoing PCI. They were divided randomly into two groups depending on whether PercuSurge was used (n=30) or not used (control n=37) during PCI. Final TIMI flow, TMP grade and the time involved in or necessary for various steps of the PCI were recorded. PercuSurge showed significantly greater achievement of TIMI III flow and TMP III grade (p<0.01). Its use was associated with less total procedural time (p<0.05). The time required from guidewire crossing to stent placement; from guidewire crossing to TIMI III flow and from predilatation/stent placement to optimal TIMI flow was significantly reduced with its use (p<0.05 for all). Slow/no-reflow was significantly reduced (p<0.001), thus reducing intracoronary vasodilators and GP IIb/IIIa antagonists requirements. A 2 years' follow-up revealed four deaths in control and one death in PercuSurge group. CONCLUSION: PercuSurge reduced the total procedural time with better and faster optimal TIMI flow and TMP grade in primary/rescue PCI and was associated with less long term events. PMID- 19218718 TI - The relation between total daily caloric intake and blood pressure. AB - BACKGROUND: More caloric intake and less physical activity engenders more obesity and any degree of weight gain even to the level that is not defined as overweight is associated with an increase of hypertension. It is not defined that if total caloric intake irrespective of weight gain had any effect on blood pressure. This is an observational study about the relationships between blood pressure and daily calorie intake in 1,061 female and 890 male aged 20-51 years. METHOD: Relation of daily total caloric intake and blood pressure in normotensive and hypertensive samples. Dietary data were obtained from a food-frequency questionnaire which was analyzed by software designed for Iranian foods designed by Isfahan cardiovascular research center to calculate the total daily caloric intake. Blood pressure was measured three times by korotkoff method. The relation between daily total caloric intake and blood pressure was analyzed by general linear regression and logistic regression. RESULTS: Generalized linear regression revealed no significant relation between either systolic or diastolic blood pressure and daily total caloric intake irrespective of normal or abnormal blood pressure. This association remained non-significant even after adjustment for body mass index (BMI) p<0.05. Logistic regression analysis revealed that there is no significant relation between total caloric intake and existence of systolic or diastolic hypertension OR=1, p>0.05. After adjustment for BMI also, there was no significant relation between these two parameters OR=1, p>0.05. CONCLUSION: Usual relation between caloric intake and blood pressure in obese may be due to insulin resistance induced by obesity. So total daily caloric intake in general population had no significant effect on blood pressure and on development of hypertension when the effect of obesity is adjusted. PMID- 19218719 TI - Indo Heart Survey on latent abnormal glucose regulation in patients with coronary artery disease without diabetes across India. AB - BACKGROUND: The Euro Heart Survey on diabetes and heart has demonstrated high prevalence of latent glucose abnormalities in non-diabetic patients with coronary artery disease (CAD) in the European population. The aim of our survey was to assess the prevalence of latent abnormal glucose regulation in adult non-diabetic CAD patients in India. METHODS AND RESULTS: Seven centers distributed across India recruited 350 patients. The diagnosis of CAD was done by coronary angiography showing >50% stenosis in any major epicardial coronary artery or its branches. Oral glucose tolerance test (OGTT) and fasting glucose levels were used to characterize glucose metabolism. Venous plasma glucose was measured before (fasting) and 2 hours after ingestion of glucose. Impaired fasting glucose (IFG) was defined as OGTT (0 minute)>or=100 mg/dl but <126 mg/dl and OGTT (2 hours)<140 mg/dl. Impaired glucose tolerance (IGT) was defined as OGTT (0 minute)<126 mg/dl and OGTT (2 hours)>or=140 mg/dl but <200 mg/dl. Of the 350 patients studied, 176 (50.28%) had impaired glucose regulation (IFG-28 [8%]; IGT-148[42.28%]) and 75 (21.42%) had newly detected Diabetes. In all 251 (71.7%) patients with CAD had previously undetected abnormal glucose regulation. CONCLUSION: This survey demonstrates the presence of abnormal glucose regulation in almost three quarters of the non-diabetic Indian CAD patients. OGTT should be recommended as routine screening test for detecting latent glucose abnormalities in all CAD patients. PMID- 19218720 TI - Elastic properties of aorta are impaired in patients with slow coronary flow phenomenon. AB - BACKGROUND: Increased stiffening and decreased distensibility of the large arteries are associated with the presence of coronary artery disease and has been related to increased cardiovascular mortality in different populations. AIM: Aim of this present study was to investigate the elastic properties of the aortic wall in patients with slow coronary flow phenomenon. MATERIAL AND METHOD: We studied 20 patients with slow coronary flow phenomenon (age: 40+/-12 years)and 15 normal control subjects by echocardiography. Aortic strain (%) and distensibility (10(-3) mmHg(-1)) were calculated from the echocardiographically-derived thoracic Ao diameters (mm). The measurement of pulse pressure was obtained by cuff sphygmomanometry. RESULTS: There was no difference in the left ventricular ejection fraction, left ventricular end-diastolic and end-systolic diameters, left atrial diameters, left ventricular mass index between patients with slow coronary flow phenomenon and control groups. Maximal aortic diastolic diameter was increased in patients with slow coronary flow phenomenon compared with control group (p<0.05). Ao distensibility and Ao strain were lower in the patients with slow coronary flow phenomenon compared with control group (p<0.05). CONCLUSION: Reduced thoracic aortic elastic properties in patients with slow coronary flow phenomenon,assessed by echocardiography, apart from demonstrating subclinical atherosclerosis may also contribute to the etiopathogenesis of the slow coronary flow phenomenon necessitating more aggressive primary preventive measure. PMID- 19218721 TI - Comparison between transcatheter closure and minimally invasive surgery for fossa ovalis atrial septal defect: a single institutional experience. AB - BACKGROUND: Although, conventional surgical closure of atrial septal defect (ASD) provides excellent results with very low mortality and morbidity, it leaves the scar of incision and postoperative pain. Newer treatment modalities like minimal invasive surgery and percutaneous closure are being increasingly used nowadays where available. AIM: To compare the patient population, success, safety, and efficacy of transcatheter closure of ASD (Group A) with that of minimally invasive surgery (Port Access) (Group B). METHODS: In this retrospective non randomized study, a record of a total of 640 patients with diagnosis of ASD secundum between May 1997 and October 2006 were reviewed. A total of 470 out of 640 patients were selected for transcatheter closure (Group A) while 170 patients were taken for surgical closure by minimally invasive port access surgery (Group B). The safety and efficacy of two groups was evaluated on the basis of morbidity and mortality, duration of intensive care unit (ICU) stay, total duration of hospital stay, post-procedural complications, residual sequel at time of discharge, and residual flow across the ASD. RESULTS: Success rate in two groups was 97.1% and 99.4%, respectively and had no statistically significant difference. Similarly major complication rate also had no difference in statistical significance (1.8% and 2.9% for Group A and B, respectively). Group B patients had longer hospital stay. A small but significant number of patients were not found suitable for device closure. This number is likely to decrease as experience with technique increases. Port access surgery is currently not possible in small children (femoral artery diameter 35 mm) due to difficulty in cannulation. CONCLUSION: Percutaneous device closure of ASD can be offered as a treatment option in suitable patients. Port access is minimally invasive and an equally safe and effective alternative choice in ASDs with deficient rim in patient with appropriate age and weight. PMID- 19218722 TI - Contrast-induced nephropathy. AB - Contrast-induced nephropathy (CIN) is a fairly common yet under-recognized clinical condition in the interventional cardiological practice. A 25% or more than 0.5 mg/dl rise of serum creatinine is generally accepted as defining CIN. The most important risk factors for CIN are pre-existing renal disease, volume of contrast media, nature of contrast media, and diabetes mellitus. Among the various postulated pathophysiological mechanisms for the precipitation of CIN, intra-renal vasoconstriction, and oxidative tubular injury are the best documented. Effective strategies to prevent CIN include adequate peri-procedural hydration with normal saline, use of N-acetylcysteine, keeping the volume of contrast media as low as feasible, and avoiding high-osmolal ionic contrast media. However, more efficient and cost-effective strategies are being developed and the search for the ideal contrast media is still on. PMID- 19218723 TI - Management of hypertension in heart failure: striking lack of evidence for a common problem. AB - Arterial hypertension is common either as a concomitant or pathogenetic entity in patients with systolic heart failure and in those with heart failure and normal ejection fraction. In free-living communities, more than half of the patients of heart failure with normal ejection fraction (HFnEF) have hypertension somewhat more than that occurring in presence of systolic heart failure. In acute heart failure, co-existent hypertension is much more frequent. Separate guidelines exist for management of hypertension and systolic heart failure. There are no published guidelines for management of HFnEF. There are contradictory recommendations with regard to drug management of hypertension and systolic heart failure. This review examines the available literature on this common co-existing combination and suggests some new recommendations. PMID- 19218724 TI - Successful thrombolysis of left main and LAD artery thrombus presenting as inferior ST-elevation myocardial infarction. PMID- 19218725 TI - When to suspect an aortopulmonary window in a pink Tetralogy of Fallot. PMID- 19218726 TI - A case of complete revascularization with nine sirolimus eluting stents. AB - Patients with multivessel disease and LV dysfunction are conventionally treated with coronary artery bypass surgery. With the advent of drug eluting stents, complex lesions are now being tackled percutaneously. The ARTS II registry demonstrated the efficacy and safety of sirolimus-eluting stents (SES) in patients with multivessel disease. We report a case of "complete revascularization" with 9 sirolimus-eluting stents in a patient with multivessel disease and LV dysfunction with an angiographic evaluation at 6 months and 1 year. PMID- 19218727 TI - Transcatheter closure of multiple atrial septal defects in situs inversus with dextrocardia. PMID- 19218728 TI - Tornus catheter facilitated recanalization of chronic total occlusions--another niche device for a difficult lesion subset. AB - One of the last frontiers facing interventionalists is the treatment of chronic total occlusions. Chronic total occlusions (CTO) are present in about one-third of all diagnostic coronary angiograms. Recanalizations of these CTOs have demonstrated a trend towards lower mortality. Unfortunately, use of standard angioplasty equipment has only resulted in a 50-60% success rate.5,6,7 Several recently developed guidewires have increased success rates to 79-90%. Further success in CTO revascularization has been improved by use of novel techniques such as crossing the lesion in a retrograde fashion from collateral vessels. Niche devices such as the Frontrunner CTO catheter (Lumend Inc., Johnson and Johnson, Picataway, NJ) have also facilitated in successful treatment of CTOs. Unfortunately, none of these developments address the issue of deliverability of angioplasty balloons and stents. A newly developed device, the Tornus catheter (Asahi Intech; Abbot Vascular, Redwood City, CA), plays a niche role in the recanalization of this lesion subset. The following contains two case reports which highlight successful revascularization of CTOs with utilization of this new catheter. PMID- 19218729 TI - Myo-intimal hyperplasia in renal artery stent: multi-detector CT demonstration. PMID- 19218730 TI - Spotlight on "peer-review": the noble foundation of scientific research. PMID- 19218731 TI - Reducing the burden of coronary artery disease in India: challenges and opportunities. AB - Asian Indians--living both in India and abroad--have one of the highest rates of coronary artery disease (CAD) in the world, three times higher than the rates among Caucasians in the United States. The CAD among Indians is usually more aggressive at the time of presentation compared with whites or East Asians. The overall impact is much greater because the CAD in Asian Indians affects the "younger" working population. This kind of disproportionate epidemic among the young Indians is causing tremendous number of work days lost at a time when India is experiencing a dizzying economic boom and needs a healthy populace to sustain this boom. While the mortality and morbidity from CAD has been falling in the western world, it has been climbing to epidemic proportions among the Indian population. Various factors that are thought to contribute to this rising epidemic include urbanization of rural areas, large-scale migration of rural population to urban areas, increase in sedentary lifestyle, abdominal obesity, metabolic syndrome, diabetes, inadequate consumption of fruits and vegetables, increased use of fried, processed and fast foods, tobacco abuse, poor awareness and control of CAD risk factors, unique dyslipidemia (high triglycerides, low HDL cholesterol levels), and possible genetic predisposition due to lipoprotein (a) [Lp(a)] excess. The effect of established, as well as novel, risk factors is multiplicative, not just additive (total effect>sum of parts). The management would require aggressive individual, societal, and governmental (policy and regulatory) interventions. Indians will require specific lower cut-offs and stricter goals for treatment of various risk factors than is currently recommended for western populations. To this end, the First Indo-US Healthcare Summit was held in New Delhi, India on December 14 and 15, 2007. The participants included representatives from several professional entities including the American Association of Physicians of Indian origin (AAPI), Indian Medical Association (IMA), Medical Council of India (MCI), and Government of India (GOI) with their main objective to address specific issues and provide precise recommendations to implement the prevention of CAD among Indians. The summary of the deliberations by the committee on "CAD among Asian Indians" and the recommendations are presented in this document. OBJECTIVES: Discussion of demographics of CAD in Indians-both in India and abroad, current treatment strategies, primordial, primary, and secondary prevention. Development of specific recommendations for screening, evaluation and management for the prevention of CAD disease epidemic among Asian Indians. Recommendations for improving quality of care through professional, public and private initiatives. PMID- 19218733 TI - On the sensitivity of IMRT dose optimization to the mathematical form of a biological imaging-based prescription function. AB - Voxel-based prescriptions of deliberately non-uniform dose distributions based on molecular imaging, so-called dose painting or theragnostic radiation therapy, require specification of a transformation that maps the image data intensities to prescribed doses. However, the functional form of this transformation is currently unknown. An investigation into the sensitivity of optimized dose distributions resulting from several possible prescription functions was conducted. Transformations between the radiotracer activity concentrations from Cu-ATSM PET images, as a surrogate of tumour hypoxia, and dose prescriptions were implemented to yield weighted distributions of prescribed dose boosts in high uptake regions. Dose escalation was constrained to reflect clinically realistic whole tumour doses and constant normal tissue doses. Optimized heterogeneous dose distributions were found by minimizing a voxel-by-voxel quadratic objective function in which all tumour voxels were given equal weight. Prescriptions based on a polynomial mapping function were found to be least constraining on their optimized plans, while prescriptions based on a sigmoid mapping function were the most demanding to deliver. A prescription formalism that fixed integral dose was less sensitive to errors in the choice of the mapping function than one that boosted integral dose. Integral doses to normal tissue and critical structures were insensitive to the shape of the prescription function. Planned target dose conformity improved with smaller beamlet dimensions until the inherent spatial resolution of the functional image was matched. Clinical implementation of dose painting depends on advances in absolute quantification of functional images and improvements in delivery techniques over smaller spatial scales. PMID- 19218734 TI - The geometric response function for convergent slit-slat collimators. AB - We have derived an analytic geometric transfer function (GTF) for a convergent slit-slat collimator that treats the parallel slit-slat collimator as a special case. The effective point spread function (EPSF) is then derived from the GTF through the Fourier transform. The results of these derivations give an accurate description of the complete geometric response for a slit-slat collimator that includes the effects of the shape and orientation of the slit and slats. We have also derived exact and approximate sensitivity formulae and spatial resolution formulae using the EPSF. PMID- 19218735 TI - Signal filtering algorithm for depth-selective diffuse optical topography. AB - A compact filtered backprojection algorithm that suppresses the undesirable effects of skin circulation for near-infrared diffuse optical topography is proposed. Our approach centers around a depth-selective filtering algorithm that uses an inverse problem technique and extracts target signals from observation data contaminated by noise from a shallow region. The filtering algorithm is reduced to a compact matrix and is therefore easily incorporated into a real-time system. To demonstrate the validity of this method, we developed a demonstration prototype for depth-selective diffuse optical topography and performed both computer simulations and phantom experiments. The results show that the proposed method significantly suppresses the noise from the shallow region with a minimal degradation of the target signal. PMID- 19218736 TI - A study of coronary artery rotational motion with dense scale-space optical flow in intravascular ultrasound. AB - This paper describes a novel method for estimating tissue motion in two dimensional intravascular ultrasound (IVUS) images of a coronary artery. It is based on the classical Lukas-Kanade (LK) algorithm for optical flow (OF). The OF vector field quantifies the amount of misalignment between two consecutive frames in a sequence of images. From the theoretical standpoint, two fundamental improvements are proposed in this paper. First, using a simplified representation of the vessel wall as a medium with randomly distributed scatterers, it was shown that the OF equation satisfies the integral brightness conservation law. Second, a scale-space embedding for the OF equation was derived under the assumption of spatial consistency in IVUS acquisitions. The spatial coherence is equivalent to a locally affine motion model. The latter effectively captures and appropriately describes a complex deformation pattern of the coronary vessel wall under the varying physiological conditions (i.e. pulsatile blood pressure). The accuracy of OF tracking was estimated on the tissue-mimicking phantoms subjected to the controlled amount of angular deviation. Moreover, the performance of the classical LK and proposed approach was compared using the simulated IVUS images with an atherosclerotic lesion. The experimental results showed robust and reliable performance of up to 5 degrees of rotation, which is within the plausible range of circumferential displacement of the coronary arteries. Subsequently, the algorithm was used to analyze vessel wall motion in 18 IVUS pullbacks from 16 patients. The in vivo experiments revealed that the motion of coronary arteries is primarily determined by the cardiac contraction. PMID- 19218737 TI - Denoising human cardiac diffusion tensor magnetic resonance images using sparse representation combined with segmentation. AB - Cardiac diffusion tensor magnetic resonance imaging (DT-MRI) is noise sensitive, and the noise can induce numerous systematic errors in subsequent parameter calculations. This paper proposes a sparse representation-based method for denoising cardiac DT-MRI images. The method first generates a dictionary of multiple bases according to the features of the observed image. A segmentation algorithm based on nonstationary degree detector is then introduced to make the selection of atoms in the dictionary adapted to the image's features. The denoising is achieved by gradually approximating the underlying image using the atoms selected from the generated dictionary. The results on both simulated image and real cardiac DT-MRI images from ex vivo human hearts show that the proposed denoising method performs better than conventional denoising techniques by preserving image contrast and fine structures. PMID- 19218738 TI - A method for the production of composite scintillators for dosimetry in diagnostic radiology. AB - Plastic scintillators are used in the dosimetry of photons in radiotherapy. Their use in diagnostic radiology is affected by the drop in response at lower photon energies due to inadequate composition (effective atomic number) and chemical quenching. To compensate for this deficiency, a method for the production of composite polystyrene-based plastic scintillators was devised allowing the incorporation of inorganic scintillation powder. Disks of 10 mm diameter and 1 mm thickness optimized for a flat energy response referred to kerma in air were produced using CaWO(4), ZnS:Ag and CaF(2):Eu as additives. In an HVL range of 2.26-13.69 mmAl, the response was within an interval of +/-2.8% for CaF(2):Eu as additive, +/-3.2% for CaWO(4) and +/-10.9% for ZnS:Ag, respectively. The response of a commercial plastic scintillator (BC470) stays within +/-13.6%. The temperature dependence of the composite scintillator using CaF(2):Eu is lowest with a variation of +3.7% to -3.6% in an interval from 5 degrees C to 45 degrees C. The deficiency in photon absorption at lower energies due to the effective atomic number is reduced but not fully compensated by the additive scintillators. The optimized concentrations were established for the scintillator dimensions used. PMID- 19218739 TI - Calculation of water equivalent thickness of materials of arbitrary density, elemental composition and thickness in proton beam irradiation. AB - In proton therapy, the radiological thickness of a material is commonly expressed in terms of water equivalent thickness (WET) or water equivalent ratio (WER). However, the WET calculations required either iterative numerical methods or approximate methods of unknown accuracy. The objective of this study was to develop a simple deterministic formula to calculate WET values with an accuracy of 1 mm for materials commonly used in proton radiation therapy. Several alternative formulas were derived in which the energy loss was calculated based on the Bragg-Kleeman rule (BK), the Bethe-Bloch equation (BB) or an empirical version of the Bethe-Bloch equation (EBB). Alternative approaches were developed for targets that were 'radiologically thin' or 'thick'. The accuracy of these methods was assessed by comparison to values from an iterative numerical method that utilized evaluated stopping power tables. In addition, we also tested the approximate formula given in the International Atomic Energy Agency's dosimetry code of practice (Technical Report Series No 398, 2000, IAEA, Vienna) and stopping power ratio approximation. The results of these comparisons revealed that most methods were accurate for cases involving thin or low-Z targets. However, only the thick-target formulas provided accurate WET values for targets that were radiologically thick and contained high-Z material. PMID- 19218740 TI - Biventricular noncompaction associated with left ventricular systolic and diastolic dysfunction and severe pulmonary hypertension in a young man. AB - Noncompaction of the ventricular myocardium is a recently recognized genetic cardiomyopathy. The left ventricle is the most affected site, but right ventricular involvement has been reported in some cases. Diagnosis is made with 2 dimensional echocardiography or cardiac magnetic resonance imaging. The major clinical manifestations are heart failure, arrhythmias and embolic events. A 20 year old man had left and right ventricular noncompaction complicated by severe pulmonary hypertension, which is one of the first cases of biventricular noncompaction associated with severe pulmonary hypertension. Pulmonary hypertension may be a consequence of increased pulmonary venous pressures caused by systolic and diastolic heart dysfunction secondary to noncompaction. PMID- 19218741 TI - Successful percutaneous intervention to acute myocardial infarction presenting with typical chest pain in transplanted heart. AB - Allograft coronary artery disease (CAD) is the main limiting factor of the long term survival in cardiac transplant recipients. It is present in approximately 50% of patients at 5 years post-transplant. Allograft CAD might result in congestive heart failure, ventricular arrhythmias and sudden cardiac death. Acute coronary syndrome is a rare presentation of allograft CAD, because symptoms are usually atypical or absent as a result of cardiac denervation. However, it should be reminded that reinnervation of the transplanted heart might occur, and angina pectoris could be felt as a sign of an acute coronary syndrome. We present a 27 year-old female cardiac transplant patient with an acute myocardial infarction, who was successfully treated with percutaneous transluminal coronary angioplasty. PMID- 19218742 TI - Insufficient membrane fusion in dysferlin-deficient muscle fibers after heavy-ion irradiation. AB - Recently, SJL/J mice have been used as an animal model in studies of dysferlinopathy, a spectrum of muscle diseases caused by defects in dysferlin protein. In this study we irradiated muscle fibers isolated from skeletal muscle of SJL/J mice with heavy-ion microbeam, and the ultrastructural changes were observed by electron microscopy. The plasma membrane of heavy-ion beam irradiated areas showed irregular protrusions and invaginations. Disruption of sarcomeric structures and the enhancement of autophagy were also observed. In addition, many vesicles of varying size and shape were seen to be accumulated just beneath the plasma membrane. This finding further supports the recent hypothesis that dysferlin functions as a membrane fusion protein in the wound healing system of plasma membrane, and that the defect in dysferlin causes insufficient membrane fusion resulting in accumulation of vesicles. PMID- 19218743 TI - [cAMP signaling for ERK activation in neuronal cells]. PMID- 19218744 TI - [EGFR and VEGF-targeted drugs: clinical evidence for their effects from evaluation of biomarkers]. PMID- 19218745 TI - [New drug targets and research methods for androgenetic alopecia]. PMID- 19218746 TI - [The current treatments and perspectives for androgenetic alopecia]. PMID- 19218747 TI - [Drug-induced testicular toxicity]. PMID- 19218748 TI - [Toxicology of the respiratory system]. PMID- 19218749 TI - [Pharmacological and clinical profiles of the anticoagulant enoxaparin sodium (Clexane 2000 IU S.C. Inj. Kit)]. PMID- 19218750 TI - [Pharmacological profiles and clinical effects of oral sustained release beraprost sodium (Careload LA tablets 60 microg, Berasus LA tablets 60 microg)]. PMID- 19218751 TI - Seoul Declaration on Safety and Health at Work. PMID- 19218752 TI - Occupational exposure to whole body vibration-train drivers. AB - Whole body vibration exposure of the train drivers working for State Railway Lines is assessed by referring to ISO standard 2631 -1 and EU directive 2002/44/EC. The vibration measurements were done in the cabins of suburban and intercity train drivers. Suburban train driver performs his job usually in standing posture. Whereas intercity train driver works generally in seated (bending forward) posture and exposed to longer periods of continuous vibration, compared to suburban train drivers. The mean accelerations, a, along lateral, y, and vertical, z, directions measured on the driver seat (on the cabin floor) of the intercity (suburban) train were 1.4a (y) = 0.55 (0.28) m/s(2) and a (z) = 0.65 (0.23) m/s(2). Daily exposure action values suggested in EU directive are exceeded in case of intercity train drivers and their exposure falls within the health caution zone of ISO 2631-1. Intercity train drivers are therefore under the risk of having spinal disorders. A health surveillance plan requiring every five years the reassessment of the state of the spinal system of train drivers is suggested. As an early preventive measure, extended work day or more than one shift in a day is advised to be discouraged. PMID- 19218753 TI - Alteration of brain neurotransmitters in female rat offspring induced by prenatal administration of 16 and 64 mg/kg of 2,2',4,4',5,5'-hexachlorobiphenyl (PCB153). AB - PCB153 (2,2',4,4',5,5'-hexachlorobiphenyl), a non-coplanar PCB and the congener most widely distributed in the environment, was orally administered to pregnant Sprague-Dawley (Crj: CD (SD) IGS) rats from gestation day 10 through 16 at doses of 0 (control), 16 and 64 mg/kg body weight. Female pups were sacrificed at 1, 3, 6, and 9 wk, and at 1 yr of age to evaluate the differences in brain neurotransmitters and their metabolites between PCB153-exposed and control groups. Brain levels of norepinephrine (NE), 3-methoxy-4-hydroxyphenylglycol (MHPG), dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), serotonin (5HT), 5-hydroxyindoleacetic acid (5HIAA), acetylcholine (ACh), and choline (Ch) in discrete brain regions or in whole brain were measured. At 1 to 3 wk after birth, brain levels of DA, DOPAC, HVA, 5HT and 5HIAA in PCB-exposed groups were higher than those of the control group. At 9 wk after birth, DA turnover was reduced in half of the four brain areas examined (forebrain and hindbrain), and 5HIAA levels were increased in all brain areas in the PCB-treated group compared to those of the control group. At 1 yr after birth, the levels of DA, DOPAC, and HVA in the hippocampus, hypothalamus, and medulla oblongata were lower in the PCB-exposed groups than in the control group. Prenatal exposure to PCB153 stimulated the turnover of 5HT neurons in the brain of female offspring at early stages (1 to 9 wk) of development. On the other hand, the turnover of DA neurons in the PCB-exposed groups was reduced in late stages (9 wk to 1 yr) of development compared with that of the control group. The brain neurotransmitters of dams treated with PCB were assayed at 3 wk after delivery (15 wk old), and decreases in DA, DOPAC, and HVA were observed. PCB153 reduced the activity of DA neurons in the brain of dams. These results are discussed in relation to health effects observed in humans exposed to PCBs. PMID- 19218754 TI - Working hours, coping skills, and psychological health in Japanese daytime workers. AB - This study examined the relationship between coping skills, working hours, and psychological health among Japanese daytime workers. Self-administered questionnaires were mailed to a randomly selected sample of 2,000 workers who were members of a pre-recruited market research panel. A total of 1,821 participants responded (response rate=91.1%). Participants completed a questionnaire regarding working hours, coping skills, and psychological health (negative emotions, fatigue, and concentration/activity levels). Analyses of covariance were conducted to determine the relations of number of working hours, coping skills, and their interactions to psychological health with control for sex, age, drinking, job type, and employment type. Results revealed that working hours were significantly associated with fatigue and concentration/activity levels. High levels of instrumental support and positive reframing were significantly associated with low levels of negative emotions, fatigue, and concentration/activity levels. High levels of self-blame, denial, substance use, venting, self-distraction, religion, and behavioral disengagement were significantly associated with high levels of negative emotions, fatigue, and concentration/activity levels. This study suggests that improving coping skills such as using instrumental support or positive reframing may reduce the adverse health effects of long working hours. PMID- 19218756 TI - Evaluation of occupational exposures to respirable dust in underground coal mines. AB - Dust can be produced by almost all mining operations in underground coal mines and seen all around the mine. Different occupational groups get exposed to different dust levels and in order to minimize the probability of developing coal workers' pneumoconiosis (CWP), it is necessary to investigate the workers exposed to dust. This study aimed to evaluate the dust concentration conditions in underground coal mines and also the occupational health risks associated with exposures to respirable dust. The data obtained from the dust measurement studies conducted in various underground coal mines between the years 1978-2006 was evaluated by using analysis of variance (ANOVA) and Tukey-Kramer procedure. In the statistical analyses, the comparison of dustiness between mines and their mining regions were made by using the average dust concentration values. In addition, the numbers of workers with doubtful pneumoconiosis diagnosis were evaluated according to the occupational job category by using the number of the workers with the job illness. It was concluded that the production regions have the higher dust concentration levels and the CWP is mostly diagnosed in the workers working in production regions. PMID- 19218755 TI - Exposure of jeepney drivers in Manila, Philippines, to selected volatile organic compounds (VOCs). AB - The objective of this study was to assess the occupational exposure of jeepney drivers to selected volatile organic compounds (VOCs) in Manila, Philippines. Personal sampling was conducted on 15 jeepney drivers. Area sampling was conducted to determine the background VOC concentration in Manila as compared to that in a rural area. Both personal and area samples were collected for 5 working days. Samples were obtained using diffusive samplers and were analyzed for 6 VOCs (benzene, toluene, ethylbenzene, m,p-xylene and o-xylene) using gas chromatography. Results showed that the average personal exposure concentration of jeepney drivers was 55.6 (+/-9.3), 196.6 (+/-75.0), 17.9 (+/-9.0), 72.5 (+/ 21.1) and 88.5 (+/-26.5) microg/m(3) for benzene, toluene, ethylbenzene, m,p xylene and o-xylene, respectively. The urban ambient concentration was 11.8 (+/ 2.2), 83.7 (+/-40.5) and 38.0 (+/-12.1) microg/m(3) for benzene, toluene and o xylene, respectively. The rural ambient concentration was 14.0 (+/-6.0) and 24.7 (+/-11.9) microg/m(3) for toluene and o-xylene, respectively. The personal samples had significantly higher (p<0.05) concentrations for all selected VOCs than the urban area samples. Among the area samples, the urban concentrations of benzene and toluene were significantly higher (p<0.05) than the rural concentrations. The personal exposures for all the target VOCs were not significantly different among the jeepney drivers. PMID- 19218757 TI - First Korean Working Conditions Survey: a comparison between South Korea and EU countries. AB - This report aims to provide an overview of the intensity of work and work satisfaction in Korea by comparing the first Korean Working Conditions Survey with the results of the fourth European Working Conditions Survey in 2005. According to the comparison, Korean workers work longer hours than workers in the majority of EU countries. However, work intensity in the EU 27 was 1.5 times stronger than Korea. In Korea, the rate of violence, bullying and harassment in the workplace is much lower than the EU 27, but in terms of work satisfaction, EU workers responded more positively than Korean workers. The amount of leave in Korean workers was much lower than the EU counterparts, yet the average number of accident-related or other work-related leaves was greater in Korean workers than workers in the EU. The pattern of exposure to physical factors, namely the most frequent hazards, such as, repetitive movement and painful or tiring posture, is similar in Korean and EU workers. However, the proportion of each exposure is larger in Korean workers than EU workers. In Korea, the most frequently reported work-related symptoms are musculoskeletal disorders followed by stress and fatigue. These patterns are quite similar in Korea and EU countries. PMID- 19218758 TI - Stress at work among electric utility workers. AB - In the last decades there was an increase in stress at work and its effects on workers' health. These issues are still little studied in the electric utility sector. This study aims to evaluate factors associated with stress at work and to verify its associations with health status among workers of an electric company in Sao Paulo State, Brazil. A cross-sectional study was conducted with 474 subjects (87.5% of the eligible workers). Data were collected using self-reported questionnaires. A descriptive analysis, a multiple linear hierarchical regression analysis and a correlation analysis were performed. The majority of participants were males (91.1%) and the mean age was 37.5 yr. The mean score of stress level was 2.3 points (scale ranging from 1.0 to 5.0). Hierarchical multiple analyses showed that: regular practice of physical activities (p=0.025) and individual monthly income (p=0.002) were inversely associated with stress level; BMI was marginally associated with the stress level (p=0.074). The demographic characteristics were not associated with stress. Stress at work was significantly associated with physical and mental health status (p<0.001). To improve health of electric utility workers, actions are suggested to decrease stress by remuneration and an appropriate practice of physical activity aiming reduction of BMI. PMID- 19218759 TI - Respiratory toxicity of raw materials used in ceramic production. AB - The main purpose of this study was to assess the pulmonary reactions associated with exposure to raw materials used in ceramic production (RMCP). This was a cross sectional study in which 33 male workers with current exposure to RMCP and 20 healthy male unexposed workers (referent group) were interviewed and respiratory symptom questionnaires were administered to them. Furthermore, they underwent chest X-ray and lung function tests. Additionally, personal dust monitoring for airborne inhalable and respirable dust was carried out at dusty areas of the industry. To determine the chemical composition, possible silica phases and SiO(2) contents of dust samples, they were analyzed by both X-ray diffraction (XRD) and X-ray fluorescence (XRF) techniques. Demographic and socioeconomic variables of both groups were similar, except that referent individuals were, to some extent, older and heavier than their exposed counterparts. Personal dust monitoring showed that the concentrations of inhalable and respirable dust were very high and dust contained large amounts of crystalline silica. Additionally, respiratory symptom questionnaires revealed that exposed workers, compared to their unexposed counterparts, had higher prevalences of cough, wheezing, phlegm and shortness of breath. Likewise, significant decrements in some parameters of pulmonary function were noted and most of the exposed subjects showed abnormalities in their chest radiographs. These data provide further evidence in favor of the notion that exposure to RMCP, probably due to their silica contents, is associated with respiratory symptoms, radiographic abnormalities and functional impairments. PMID- 19218760 TI - Safety climate and motivation toward patient safety among Japanese nurses in hospitals of fewer than 250 beds. AB - A questionnaire survey was done to examine the associations between the dimensions of safety climate and the nurses' motivation toward patient safety. Excluding nursing directors, a total of 371 full-time female Japanese nurses (registered nurses and licensed practical nurses) working in 6 hospitals of fewer than 250 beds were analyzed. The average age was 35.6 yr (range, 20-69). For the safety climate, factor analysis (the principal factor method and promax rotation) was performed, and factors with an eigenvalue of>or=1 were extracted. Seven dimensions were extracted by factor analysis as follows: "Opportunities for nursing education", "Reporting", "Fatigue reduction", "Superiors' attitudes", "Nursing conditions", "Communications with physicians", and "Relationships among nurses". We subsequently examined the associations between the dimensions of the safety climate and "Nurses' motivation to prevent mistakes". Multiple linear regression analysis showed that "Nurses' motivation to prevent mistakes" was significantly associated with "Reporting", "Nursing conditions", and "Communications with physicians". It is necessary to improve the reporting system in hospitals. It is also necessary to improve on reducing improper communications with physicians. And appropriate nursing conditions must be actualized and maintained. PMID- 19218761 TI - Comparative sensitivity of outcome variables of a software-based Behavioral Sleep Resistance Task. AB - Unintentional sleep and performance impairment due to extended wakefulness are often the cause of traffic and work accidents. Therefore, large-scale screening instruments assessing the ability to resist falling asleep during monotonous tasks are needed. The current widespread computer use in industrial and home settings offers the possibility to use software-based instruments as an alternative for expensive hardware interfaces to assess sleep resistance. Therefore, we propose a software-based sleep resistance task based on the procedure of the Oxford SLEep Resistance test (OSLER): the Behavioral Sleep Resistance Task (BSRT). In order to validate this instrument we submitted 36 healthy individuals to four 40-min sleep resistance challenges during a night of extended wakefulness (14, 16, 18 and 20 h). Subjective sleepiness was assessed by means of the Karolinska Sleepiness Scale and a Visual Analogue Scale for sleepiness/alertness and objective sleepiness by means of a 4-min EEG-monitored fixation task. Our results show significant correlations of BSRT hit ratio, error profiles and sleep onset latency variables with subjective sleepiness and of BSRT hit ratio and 3-6 error profiles with High-Beta Central EEG activity. Additionally, using a distribution- and scale-free sensitivity index, we found that subjective sleepiness measures are the most sensitive to the effects of sustained wakefulness, closely followed by the BSRT hit ratio and BSRT error profile 3-6. PMID- 19218762 TI - VDT screen height and inclination effects on visual and musculoskeletal discomfort for Chinese wheelchair users with spinal cord injuries. AB - This study investigated the visual fatigue and musculoskeletal discomfort for wheelchair users in a VDT data entry task. Eight Chinese male wheelchair users were recruited as participants to perform 9 different experimental combinations. Three screen heights and three screen inclinations were evaluated. Visual fatigue was measured using the change in critical fusion frequency (CFF), near point accommodation (NPA) and a visual complaint questionnaire. Musculoskeletal discomfort was estimated using the rating of perceived exertion (RPE). Each participant performed a data-entry task within a 30 min period using random orders. All participants during the data-entry operation used the upright posture with the participant's lower and upper arms forming right angles. The results showed that the screen height and screen inclination significantly affected visual fatigue and musculoskeletal discomfort. On average, the optimal screen height choice for wheelchair users was at eye-level height or 5 cm below eye level with a vertical screen inclination. PMID- 19218763 TI - Tracer gas evaluations of push-pull ventilation system performance. AB - A push-pull ventilation system is effective for hazardous material exhaustion. Although a push-pull ventilation system has advantages over a local exhaust hood, some laborious adjustments are required. The pertinence of the adjustments is uncertain because it is difficult to evaluate the performance of a push-pull ventilation system quantitatively. In this study, a measurement of the capture efficiency of a push-pull ventilation system was carried out by means of a tracer gas method. The capture efficiency decreased to 39.3-78.5% when blockage material, a dummy worker and a cross draft, were set in the ventilation zone, but the efficiency was 95.1-97.9% when the cross draft was stopped. The results suggest that the uniform flow of a push-pull ventilation system will detour a blockage and the performance of the system will not be reduced unless a cross draft disturbs the uniform flow. PMID- 19218764 TI - Occupational lead exposure: blood lead levels of apprentices in Bursa, Turkey. AB - The aim of this cross-sectional study was to compare the blood lead levels of apprentices working in lead-related industries with those of non-apprentice controls and to evaluate the influence of such occupational exposure. The study was conducted between October 2004 and June 2005 in Bursa, Turkey. Subjects included 231 apprentices and 252 male controls, age ranging from 14-19 yr old. Inductively coupled plasma mass spectrometry (ICP-MS) was used to analyze blood lead levels. The overall mean blood lead level was 3.62 microg/dl (95% CI 3.39 3.85), the apprentice mean was 4.99 microg/dl (95% CI 4.60-5.38), and the control group mean was 2.37 microg/dl (95% CI 2.22-2.51). The blood lead level of apprentices was significantly higher than that of controls (p<0.05). Results show that occupational exposure increases the blood lead levels of apprentices, although the impact on their immediate health is low. Measurement of blood lead levels of apprentices should be taken periodically, and a regular medical surveillance program should be established. PMID- 19218765 TI - Can we predict a prolonged course and intractable cases of herpes simplex encephalitis? PMID- 19218766 TI - Comparative performance of three metabolic syndrome definitions in the prediction of acute coronary syndrome. AB - OBJECTIVE: The value of the recently introduced definitions of metabolic syndrome (MetS) in the identification of high cardiovascular risk subjects remains questionable. We examined the association among different definitions of MetS, and the occurrence of a first-ever acute coronary syndrome (ACS). METHODS: We studied 211 patients with a first-ever ACS and 210 control subjects. We recorded cardiovascular risk factors and the presence of MetS using 3 different definitions, according to the National Cholesterol Education Program Adult Treatment Panel (NCEP-ATP) III, National Heart, Lung and Blood Institute/American Heart Association (NHLBI/AHA), and International Diabetes Federation (IDF), respectively. The association of MetS with ACS was assessed using univariate and multivariate logistic regression models after adjusting for potential confounding factors, such as gender, age, body mass index, hypertension, diabetes mellitus, and lipids. RESULTS: ACS cases had a prevalence of metabolic syndrome according to NCEP-ATP III, NHLBI/AHA, and IDF criteria of 72.5%, 81.2%, and 79.1%, respectively. The unadjusted odds ratio (OR) for a first-ever ACS were 2.32 (95% CI: 1.53-3.52, p=0.001), 2.82 (95% CI: 1.79-4.43, p=0.001), and 3.26 (95% CI: 2.12-5.00, p=0.001) for NCEP-ATP III, NHLBI/AHA, and IDF MetS definitions, respectively. Multivariate analyses revealed that only IDF-defined MetS was significantly associated with ACS (OR: 2.23 95% CI: 1.30-3.82, p=0.003), while of the MetS components only waist circumference remained independently associated with ACS (O.R: 1.045 95% CI: 1.014-1.078, p=0.005). CONCLUSION: The definition of MetS according to the IDF criteria appears to be a better predictor of ACS than NCEP-ATP III and NHLBI/AHA. PMID- 19218767 TI - Lectin-like oxidized LDL receptor-1 (LOX-1) expression in the tubulointerstitial area likely plays an important role in human diabetic nephropathy. AB - OBJECTIVE: Diabetic nephropathy (DN) is a common cause of end-stage renal disease. However, the precise mechanism of DN, which involves the role of lipid, is still not fully understood. Lectin-like oxidized LDL receptor-1 (LOX-1) is a type II single-transmembrane protein that binds oxidized low density lipoprotein (Ox-LDL). This study examined the expression of LOX-1 mRNA in renal tissues from type 2 diabetes patients with DN using in situ hybridization (ISH). PATIENTS AND METHODS: Renal tissues were obtained from 15 type 2 patients with DN and 5 minimal change nephrotic syndrome (MCNS), membranous nephropathy (MN) and normal human kidney (NHK). Glomerular and tubulointerstitial LOX-1 mRNA expression was evaluated by ISH. Results The cells positive for LOX-1 mRNA were identified in the glomeruli of DN, MCNS, MN and NHK, however, there was no positive signal in the tubulointerstitial area in MCNS and NHK. Some cells positive for LOX-1 mRNA were detectable in the tubulointerstitial area in DN and MN. In the results of glomerular staining, there was no significant difference between them. There was a significant correlation between the tubulointerstitial LOX-1 expression and the degree of the tubulointerstitial damage and urinary protein in DN. CONCLUSION: Increased expression of LOX-1 mRNA in the tubulointerstitial area may be closely linked to the development and progression of human DN, and in particular the tubulointerstitial damage. PMID- 19218768 TI - Clinical utility of serum beta-D-glucan and KL-6 levels in Pneumocystis jirovecii pneumonia. AB - OBJECTIVE: New serum markers (1-->3) beta-D-glucan (beta-D-glucan) and KL-6 are reported to be useful for the clinical diagnosis of Pneumocystis jirovecii pneumonia (PCP). However, the utility of these markers in PCP with HIV infection (HIV PCP) and without HIV (non-HIV PCP) is unknown. This study was aimed to evaluate the utility of beta-D-glucan and KL-6 for the diagnosis of PCP in patients with HIV infection (HIV PCP) and non-HIV PCP. METHODS: Retrospective study. PATIENTS: We reviewed the medical records of consecutive 35 patients. The serum levels of beta-D-glucan and KL-6 in HIV PCP and non-HIV PCP were comparatively evaluated. We evaluated these markers in survivors and non survivors. RESULTS: The detection rates of serum beta-D-glucan and KL-6 levels in non-HIV PCP were lower than those in HIV PCP (88% vs. 100%, 66% vs. 88%, respectively). The false positive rates of these markers in both groups were similar (12%, 37%, respectively). Oxygenation index, serum albumin, and mechanical ventilation were the variables which were significantly associated with poor outcome in the univariate analysis. CONCLUSION: In conclusion, beta-D glucan was a reliable diagnostic marker for PCP. However, the detection rate of beta-D-glucan and KL-6 in non-HIV PCP was lower than in HIV PCP. Neither beta-D glucan nor KL-6 was associated with the outcome of PCP. PMID- 19218769 TI - Class III beta-tubulin expression in tumor cells is correlated with resistance to docetaxel in patients with completely resected non-small-cell lung cancer. AB - OBJECTIVE: To assess the relationship between in vitro chemosensitivity evaluated by the histoculture drug response assay (HDRA) and the expression of beta-tubulin isotypes in tumors of patients with completely resected NSCLC in order to determine the predictive value of beta-tubulin in chemotherapy for NSCLC. METHODS: Expression of beta-tubulin isotypes was immunohistochemically analyzed in a series of 58 tumor samples from patients with completely resected NSCLC. The sensitivity of individual tumors to anticancer agents was evaluated by HDRA. RESULTS: Class III beta-tubulin expression by tumor cells was significantly correlated with resistance to docetaxel (p=0.0250), but not related with resistance to gemcitabine. Patient characteristics (age, gender, histology, and stage) were not associated with class III beta-tubulin expression. CONCLUSION: An abundance of class III beta-tubulin in tumor cells could be a biomarker for resistance to docetaxel in patients with completely resected NSCLC. PMID- 19218770 TI - Skeletal muscle MRI in complex regional pain syndrome. AB - BACKGROUND: Conventional magnetic resonance imaging (MRI) sequences of patients with complex regional pain syndrome (CRPS) have shown abnormal signals in skin, soft tissue, joints, bone, and bone marrow, but not yet in skeletal muscles, during the acute phase. The aim of this study was to clarify whether or not the affected muscles in CRPS patients show abnormal MRI signal intensities or signal enhancement by gadolinium dimeglumine during the acute phase. PATIENTS AND METHODS: MRI studies of skeletal muscles were performed on three patients of CRPS. Out of three patients, MRI was performed on three at stage 1, one in improving phase, two in remission phase, and one at stage 3. MRI was performed in the transaxial plane with both T2-weighted imaging (T2WI) and fat-suppressed T1 weighted imaging (T1WI) with or without gadolinium dimeglumine enhancement. RESULTS: All patients at stage 1 showed hyperintense muscle signals on T2WI and gadolinium dimeglumine enhancement on T1WI. Following clinical improvement, the hyperintense lesions reverted to near normal. Muscles in the chronic phase showed high signals on both T2WI and T1WI without gadolinium dimeglumine enhancement. CONCLUSION: MRI abnormalities in the acute phase are consistent with muscular edema, interstitial edema, and vascular hyperpermeability. These MRI findings suggest the presence of hemodynamic abnormalities caused by microangiopathy, sympathetic abnormalities, or both, which may lead to ischemia of affected muscles. Chronic phase abnormalities indicated the presence of muscle atrophy and fibrosis or fatty infiltration of the affected muscle. PMID- 19218771 TI - Chest pain without significant coronary stenosis after implantation of sirolimus eluting stents. AB - We encountered a case of exercise-induced chest pain after the implantation of sirolimus-eluting stents (SESs). She had no history of previous chest pain, and an exercise stress test just after the implantation of the SESs was negative without any symptoms. However, six months after the implantation of the SESs, she began to experience frequent episodes of severe chest pain on effort in spite of there being no significant coronary stenosis. Interestingly, severe coronary vasoconstriction was induced by an intracoronary administration of acetylcholine, and exercise stress testing revealed positive findings with chest pain and ST-T segment depression on ECG. An intensive treatment with two types of calcium channel blockers could readily and completely abolish the exercise-induced chest pain and ST-T segment depression on the ECG. In view of these findings, we presumed that coronary microvessel dysfunction and/or exercise-induced coronary vasoconstriction leading to myocardial ischemia had appeared 6 months after the implantation of the SESs. Although the pathogenesis of this phenomenon could not be completely elucidated, the anatomical and functional abnormalities of the coronary arteries associated with the implantation of the SESs may have been one of the most important mechanisms. PMID- 19218772 TI - Primary aldosteronism associated with severe rhabdomyolysis due to profound hypokalemia. AB - A 55-year-old Japanese man was admitted to our hospital with severe weakness. Without measurement of serum electrolyte concentrations, diuretic therapy for hypertension was started 2 weeks prior to admission. Laboratory findings showed profound hypokalemia (1.4 mEq/L), and extreme elevation of the serum creatinine phosphokinase levels (15,760 IU/L), suggesting that the patient had hypokalemic paralysis and hypokalemia-induced rhabdomyolysis. Further evaluations, including adrenal venous sampling strongly suggested that he had primary aldosteronism. He was treated successfully by laparoscopic adrenalectomy. This case provides an important lesson that serum electrolyte concentrations should be measured in hypertensive patients before the administration of antihypertensive agents. PMID- 19218773 TI - Spontaneous remission of diabetes insipidus due to CNS sarcoidosis. AB - Central nervous system (CNS) sarcoidosis is a crucial disease and has a poor prognosis. A 58-year-old woman had acute development of polydipsia and polyuria. Her pituitary MRI demonstrated a swelling of pituitary gland and hypophyseal stalk. She was diagnosed as central diabetes insipidus (CDI) due to CNS sarcoidosis based on the examinations and pituitary MRI findings as well as a result of cutaneous biopsy. Uveitis and bilateral hilar lymphadenopathy were observed mildly throughout. However, CDI and pituitary MRI findings were getting recovered spontaneously without steroid treatment in a couple of months, suggesting an atypical clinical course of CNS sarcoidosis. PMID- 19218774 TI - Hypothyroid Hashimoto's thyroiditis with scintigraphic and color flow doppler sonography features mimicking a hot nodule. AB - We report a 59-year-old woman who had Hashimoto's thyroiditis (HT) with hypothyroidism. A solid hypervascularized nodule in the right lobe was detected by color flow doppler sonography (CFDS). Thyroid (99m)Tc pertechnetate scintigraphy revealed a hot area in the right lobe. After three months, thyroid function tests also revealed hypothyroidism and (131)I scintigraphy was similar to the previous scintigraphy. No nodular or hypervascularized lesion in the right lobe could be identified at the sixth month of L-T4 substitution therapy. In conclusion, HT may present as a single hot nodule and hypothyroidism. Imaging findings of HT should be carefully evaluated for the precise diagnosis. PMID- 19218775 TI - Endothelial cyst of the adrenal gland associated with adrenocortical adenoma: preoperative images simulate carcinoma. AB - A 68-year-old woman was referred for characterization of a left adrenal incidentaloma. Endocrinological examinations indicated subclinical Cushing's syndrome, whereas the large volume (10 cm in diameter) and heterogeneous configuration of the tumor raised a strong suspicion of adrenal carcinoma. Hence, left adrenalectomy was performed. Histopathologically, this lesion was a thick hyaline-walled endothelial cyst, flanked with a compressed adrenocortical adenoma. The puzzling image resemblance of a variation of adrenal cyst to carcinoma necessitated histological examination for confirmative diagnosis. This is the first reported case of adrenal endothelial cyst associated with adrenocortical adenoma, the former of which alone is a rarity. PMID- 19218776 TI - Acute encephalomyelitis associated with acute viral hepatitis type B. AB - We describe the case of a 36-year-old woman who developed acute encephalo myelitis after acute viral hepatitis type B. She was admitted to the hospital with a history of general malaise and nausea of 5 days duration. Her serum showed high transaminase levels and positive HBs-Ag and increased IgM HBc-Ab titers. She had urinary dysfunction, myoclonus and postural tremor of her extremities. Several days later, she developed bilateral limb ataxia and alteration of consciousness. The cerebrospinal fluid examinations showed pleocytosis and increased protein. Treatment with high-dose methylprednisolone resulted in a marked improvement of the clinical and CSF examination. Magnetic resonance imaging of the brain and the spinal cord did not disclose abnormal lesions. The symptoms and clinical course were quite similar to those of acute disseminated encephalomyelitis. PMID- 19218777 TI - A case of intimal sarcoma of the pulmonary artery treated with chemoradiotherapy. AB - We report on a 45-year-old woman with intimal sarcoma of the pulmonary artery. She presented with a chief complaint of shortness of breath. Computed tomography (CT) of the chest showed an intraluminal hypoattenuated area extending from the main pulmonary artery into the right main pulmonary artery and bilateral lobar pulmonary arteries. She underwent resection of the lobulated mass from the pulmonary artery. The tumor was diagnosed as an intimal sarcoma. Although she received chemotherapy with amrubicin and carboplatin when the tumor recurred, the tumor enlarged. After radiotherapy was performed, CT of the chest showed shrinkage of the tumor and the regression of consolidation and ground-glass opacity. Radiotherapy and chemotherapy are treatment option for patients with pulmonary artery sarcoma. PMID- 19218778 TI - Analysis of p53 dependent damage response in sperm-irradiated mouse embryos. AB - Ionizing radiation activates a series of DNA damage response, cell cycle checkpoints to arrest cells at G1/S, S and G2/M, DNA repair, and apoptosis. The DNA damage response is thought to be the major determinant of cellular radiosensitivity and thought to operate in all higher eukaryotic cells. However, the radiosensitivity is known to differ considerably during ontogeny of mammals and early embryos of mouse for example are much more sensitive to radiation than adults. We have focused on the radiation-induced damage response during pre implantation stage of mouse embryo. Our study demonstrates a hierarchy of damage responses to assure the genomic integrity in early embryonic development. In the sperm-irradiated zygotes, p53 dependent S-phase checkpoint functions to suppress erroneous replication of damaged DNA. The transcription-dependent function is not required and the DNA-binging domain of the protein is essential for this p53 dependent S-phase checkpoint. p21 mediated cleavage arrest comes next during early embryogenesis to prevent delayed chromosome damage at morula/ blastocyst stages. Apoptosis operates even later only in the cells of ICM at the blastocyst stage to eliminate deleterious cells. Thus, early development of sperm-irradiated embryos is protected at least by three mechanisms regulated by p53 and by p21. PMID- 19218779 TI - The yield, processing, and biological consequences of clustered DNA damage induced by ionizing radiation. AB - After living cells are exposed to ionizing radiation, a variety of chemical modifications of DNA are induced either directly by ionization of DNA or indirectly through interactions with water-derived radicals. The DNA lesions include single strand breaks (SSB), base lesions, sugar damage, and apurinic/apyrimidinic sites (AP sites). Clustered DNA damage, which is defined as two or more of such lesions within one to two helical turns of DNA induced by a single radiation track, is considered to be a unique feature of ionizing radiation. A double strand break (DSB) is a type of clustered DNA damage, in which single strand breaks are formed on opposite strands in close proximity. Formation and repair of DSBs have been studied in great detail over the years as they have been linked to important biological endpoints, such as cell death, loss of genetic material, chromosome aberration. Although non-DSB clustered DNA damage has received less attention, there is growing evidence of its biological significance. This review focuses on the current understanding of (1) the yield of non-DSB clustered damage induced by ionizing radiation (2) the processing, and (3) biological consequences of non-DSB clustered DNA damage. PMID- 19218780 TI - Effect of N-acetylcysteine on radiation-induced genotoxicity and cytotoxicity in rat bone marrow. AB - The aim of this study is to evaluate the potential radioprotective effects of N acetylcysteine (NAC) against genotoxicity and cytotoxicity. The effect of WR 2721, as a representative of clinically used radioprotector, was compared with that of NAC, using the chromosomal aberration (CA) and micronucleus (MN) test systems in the irradiated rat's femoral bone marrow cells. We also investigated the mitotic index (MI), and the ratio of polychromatic erythrocytes (PCEs) to normochromatic erythrocytes (NCEs). The rats (n = 16) were divided randomly and equally into four groups: Control (C), Radiation (R), R+NAC (received irradiation and 1000 mg/kg NAC) and R+WR-2721 (received irradiation and 200 mg/kg WR-2721) rats. All the irradiated groups received whole-body gamma irradiation as a single dose of 6 Gy. Group R showed higher CA and MN formation when compared to C. Group R showed higher frequency of MN formation when compared to both R+NAC and R+WR 2721. The mean MI and PCE/NCE ratios were lower in Group R when compared to those of Group C. The mean MI and PCE/NCE ratios of both R+NAC and R+WR-2721 groups were lower when compared to those of Group C. The MI in Group R was lower when compared to that of both R+NAC and R+WR-2721 groups. In this study, the results give clues about the beneficial effects of NAC against radiation-induced genotoxicity and cytotoxicity in rat bone marrow and its effect may be comparable to that observed for WR-2721. PMID- 19218781 TI - Gene expression profiles in radiation workers occupationally exposed to ionizing radiation. AB - Ionizing radiation (IR) imposes risks to human health and the environment. IR at low doses and low dose rates has the potency to initiate carcinogenesis. Genotoxic environmental agents such as IR trigger a cascade of signal transduction pathways for cellular protection. In this study, using cDNA microarray technique, we monitored the gene expression profiles in lymphocytes derived from radiation-exposed individuals (radiation workers). Physical dosimetry records on these patients indicated that the absorbed dose ranged from 0.696 to 39.088 mSv. Gene expression analysis revealed statistically significant transcriptional changes in a total of 78 genes (21 up-regulated and 57 down regulated) involved in several biological processes such as ubiquitin cycle (UHRF2 and PIAS1), DNA repair (LIG3, XPA, ERCC5, RAD52, DCLRE1C), cell cycle regulation/proliferation (RHOA, CABLES2, TGFB2, IL16), and stress response (GSTP1, PPP2R5A, DUSP22). Some of the genes that showed altered expression profiles in this study can be used as biomarkers for monitoring the chronic low level exposure in humans. Additionally, alterations in gene expression patterns observed in chronically exposed radiation workers reinforces the need for defining the effective radiation dose that causes immediate genetic damage as well as the long-term effects on genomic instability, including cancer. PMID- 19218782 TI - The role of mitochondrial superoxide anion (O2(-)) on physiological aging in C57BL/6J mice. AB - Much attention has been focused on the mitochondrial superoxide anion (O2(-)), which is also a critical free radial produced by ionizing radiation. The specific role of the mitochondrial O2(-) on physiological aging in mammals is still unclear despite wide-spread evidence that oxidative stress is involved in aging and age-related diseases. The major endogenous source of O2(-) is generated as a byproduct of energy metabolism from mitochondria. In order to better understand how O2(-)relates to metazoan aging, we have comprehensively examined age-related changes in the levels of oxidative damage, mitochondrial O2(-) production, mitochondrial antioxidant enzyme activity and apoptosis induction in key organs of an inbred mouse strain (C57BL/6J). Oxidative damage accumulated and excess apoptosis occurred in the brain, oculus and kidney with aging, but comparatively little occurred in the heart and muscle. These rates are correlated with O2(-) levels. Mitochondrial O2(-) production levels increased with aging in the brain, oculus and kidney, and did not significantly increased in the heart and muscle. In contrast to O2(-) production, mitochondrial SOD activities increased in heart and muscle, and remained unchanged in the brain, oculus and kidney with aging. These results suggest that O2(-) production has high organ specificity, and oxidative damage by O2(-) from mitochondria mediated apoptosis can lead to organ atrophy and physiological dysfunction. In addition, O2(-) from mitochondria plays a core role in physiological aging. PMID- 19218783 TI - Microdosimetry on a mini-reactor UTR-KINKI for educational uses and biological researches. AB - Microdosimetry study has been carried out at the education and research mini reactor of Kinki University (UTR-KINKI) using a tissue equivalent gas proportional counter (TEPC). The microdosimetric single event spectra for 0.5, 1, 2, 3 and 5 microm site sizes were obtained in the lineal energy range from 1 to 1000 keV/microm. Neutron and gamma-ray fractional doses were estimated from the single event spectra. The neutron dose fraction was varied from 35 to 55% for 0.5 to 5 microm site size. The averaged lineal energy, y(D), for each site size was likewise estimated and found to be dependent on the site size. The averaged lineal energy for neutron was slightly larger than that of the fission neutrons from (252)Cf, and the averaged lineal energy for gamma-ray had similar site-size dependence of 25 keV gamma-rays and 250 kV X-rays. Relative biological effectiveness was found to be 4.1 +/- 0.13 for UTR-KINKI using Tilikidis's 2 Gy response function. The estimated RBE for UTR-KINKI neutrons is quite close to the previous biological experimental value of 4.3 +/- 0.6 for micronucleated cells in gill cell of Medaka and 4.6 +/- 0.5 for induction of lymphocyte apoptosis in the thymus of ICR mice. PMID- 19218784 TI - Differential FGF ligands and FGF receptors expression pattern in frontal and parietal calvarial bones. AB - The mammalian skull vault consists mainly of 5 flat bones, the paired frontals and parietals, and the unpaired interparietal. All of these bones are formed by intramembranous ossification within a layer of mesenchyme, the skeletogenic membrane, located between the dermal mesenchyme and the meninges surrounding the brain. While the frontal bones are of neural crest in origin, the parietal bones arise from mesoderm. The present study is a characterization of frontal and parietal bones at their molecular level, aiming to highlight distinct differences between the neural crest-derived frontal and mesodermal-derived parietal bone. We performed a detailed comparative gene expression profile of FGF ligands and their receptors known to play crucial role in skeletogenesis. This analysis revealed that a differential expression pattern of the major FGF osteogenic molecules and their receptors exists between the neural crest-derived frontal bone and the paraxial mesoderm-derived parietal bone. Particularly, the expression of ligands such as Fgf-2, Fgf-9 and Fgf-18 was upregulated in frontal bone on embryonic day 17.5, postnatal day 1 and postnatal day 60 mice. Frontal bone also elaborated higher levels of Fgf receptor 1, 2 and 3 transcripts versus parietal bone. Taken together, these data suggest that the frontal bone is a domain with higher FGF signaling competence than parietal bone. PMID- 19218785 TI - Anatomy of the ligamentum venosum arantii and its contribution to the left hepatic vein and common trunk control. A study on cadaveric livers. AB - BACKGROUND: The control of the left hepatic vein (LHV) and the common trunk of the middle hepatic vein (MHV) and LHV (CT) is considered difficult during liver resection and could be improved by detailed knowledge on the ligamentum venosum Arantii (LV). AIM: The aim of this study was to describe the LV and its connections to the LHV and the CT and to present surgical relevance of the obtained data. MATERIAL AND METHODS: During autopsy of 50 cadavers of both sexes, the LV was exposed, measured and then dissected, simulating a surgical maneuver to facilitate the approach to the LHV and CT. The extrahepatic parts of the LHV, MHV and CT were measured. RESULTS: The LV was 52-70 mm long and 5-8 mm thick. It had a fibrotic structure and was not patent in 96% of the cases. The extrahepatic part of the LHV measured 3-19 mm, that of the MHV 3-18 mm and that of the CT 4-15 mm. CONCLUSION: LV dissection facilitated extraparenchymatous clamping of the hepatic veins: the extrahepatic parts of the LHV and CT measured > 3 mm in 86 and 84% of the cases, respectively. PMID- 19218786 TI - New index for analysis of polysomnography, 'integrated area of desaturation', is associated with high cardiovascular risk in patients with mild to moderate obstructive sleep apnea. AB - BACKGROUND: Although obstructive sleep apnea (OSA) severity is evaluated by the apnea-hypopnea index (AHI), the value of AHI in evaluating cardiovascular risks, especially in mild to moderate OSA, is unclear. OBJECTIVES: The purpose of this study is to evaluate the validity of a new index, the integrated area of desaturation (IAD), to detect the incidence of cardiovascular events (CVEs) in such patients. METHODS: We enrolled 230 consecutive patients with mild to moderate OSA and 354 with severe OSA diagnosed by polysomnography, of whom 53 and 112, respectively, had CVEs. The IAD was calculated by dividing the area of desaturation by total sleep time in polysomnography. C-reactive protein (CRP) was also measured for all patients. RESULTS: In the mild to moderate OSA patients, the mean IAD of the CVEs group was significantly higher than that of the non-CVE group (94.4 +/- 82.7 vs. 62.3 +/- 50.8, p = 0.001), whereas mean AHI and 3% oxygen desaturation index were similar in both groups. Multivariate analysis demonstrated that the IAD was an independent variable for CVEs (OR 1.006, 95% confidence interval 1.001-1.012, p = 0.031). Moreover, the IAD level of the high CRP group was significantly higher than that of the low CRP group (92.9 +/- 84.8 vs. 63.9 +/- 54.5, p = 0.009). There was no significant difference in AHI, IAD or other polysomnographic parameters in the severe OSA patients. CONCLUSIONS: IAD might be superior to AHI alone in the evaluation of the history of CVEs in mild to moderate OSA patients, and it deserves attention as a possible predictor of future CVEs. PMID- 19218787 TI - New Findings in PiZZ alpha1-antitrypsin deficiency-related panniculitis. Demonstration of skin polymers and high dosing requirements of intravenous augmentation therapy. AB - Panniculitis is a recognized but unusual complication of a severe deficiency of alpha1-antitrypsin (AAT), with fewer than 100 cases described to date. Like the pathogenesis of emphysema in severe PiZZ deficiency of AAT, panniculitis has been hypothesized to be an inflammatory process, possibly related to Z AAT polymer formation and to an unopposed anti-inflammatory screen in the context of deficient serum levels of AAT. The current report presents a 31-year-old woman with PiZZ AAT deficiency-associated panniculitis. Our case extends current knowledge of AAT-associated panniculitis in 2 ways: (1) we demonstrate Z-type AAT polymers in the skin, which supports the inflammatory pathogenesis of panniculitis and the potential pro-inflammatory role of polymers; (2) we show that a high dose and long-term use of intravenous augmentation therapy (90 mg/kg body weight once weekly during 3 years) can ameliorate the frequency and severity of panniculitis associated with AAT deficiency. PMID- 19218788 TI - Relationship between the CAG repeat polymorphism in the androgen receptor gene and acne in the Han ethnic group. AB - BACKGROUND: The modulatory domain of the human androgen receptor (AR) gene contains a polymorphic CAG repeat coding for a polyglutamine tract which is inversely correlated with transcriptional activity of the AR. Androgens acting through the AR play a crucial role in the pathogenesis of acne vulgaris. We therefore investigated the relationship between CAG repeat polymorphism in the AR gene and acne susceptibility. METHODS: 206 acne patients and 200 controls participated in the study. Genomic DNA was extracted from peripheral blood lymphocytes of individual patients, and the CAG repeat region was amplified by polymerase chain reaction (PCR) using fluorescence-labeled primers. Samples were then run on an ABI 377 gene scan analysis gel with an internal molecular-weight marker. Ten male samples were chosen randomly for sequencing to confirm the number of CAG repeats. The 2-sample independent t test was used to analyze the data. RESULTS: The mean number of the CAG repeat in the AR was 22.07 (14-28) in the controls and 20.61 (13-26) in the male acne group. There was a significant correlation between the CAG repeat length and male acne. No significant difference was observed between female patients and their controls. CONCLUSION: The results suggest that the AR gene CAG repeat polymorphism may be one of the candidate genetic markers for male acne susceptibility in the Han population. PMID- 19218789 TI - Oblique view dermoscopy changes regular fibrillar pattern into parallel furrow pattern. PMID- 19218790 TI - Vitamin E supplementation reduces oxidative stress in beta thalassaemia intermedia. AB - OBJECTIVE: The aim of this investigation was to study the effect of vitamin E treatment in oxidative stress of red and white cells of beta-thalassaemia intermedia patients. METHODS: Nine patients undergoing occasional transfusions (5 females/4 males), median age 39 years (range 15-74), were recruited for oral daily administration of 400 IU vitamin E for 3 months. Twenty-seven milliliters of peripheral blood was obtained before and after 3 months of treatment, and 3 months after treatment completion. In the case of transfused patients (n = 4), blood was obtained at least 30 days after transfusion. Reactive oxygen species (ROS) was measured by flow cytometry; red blood cell (RBC) reduced glutathione (GSH) was measured by dinitrothiocyanobenzene reduction, serum malondialdehyde was measured in terms of thiobarbituric acid-reactive substances (TBARS), and alpha-haemoglobin-stabilizing protein (AHSP) mRNA expression was measured by real time polymerase chain reaction of reticulocyte RNA extracts. RESULTS: beta Thalassaemia patients presented basal levels of RBC ROS, GSH and serum TBARS statistically different compared with healthy controls. However, after vitamin E administration, patients presented a significant reduction in erythrocyte RBC ROS and serum TBARS levels. In parallel, red cell GSH was significantly increased after treatment. Peripheral mononuclear cells and T lymphocytes also demonstrated a reduction in ROS production. Therefore, after treatment, no significant differences were detected comparing patients and normal controls. Three months after treatment completion, all measurements showed a tendency of returning to basal values. A significant reduction in reticulocyte number was observed after vitamin E treatment. Vitamin E treatment did not modify levels of haemoglobin or AHSP mRNA expression. CONCLUSION: Although vitamin E is not capable of reducing anaemia in these patients, it could be useful for reducing oxidative damage in other target organs of beta-thalassaemic patients. Finally, this is the first study to analyse the effects of vitamin E on ROS production in red and white blood cells and AHSP mRNA expression. PMID- 19218791 TI - 25-Hydroxyvitamin D deficiency and inflammation and their association with hemoglobin levels in chronic kidney disease. AB - BACKGROUND/AIMS: There is limited information about the association of anemia with 25-hydroxyvitamin D [25(OH)D] deficiency and inflammation in the setting of decreased kidney function in the general population. METHODS: We examined the association that anemia has with 25(OH)D deficiency and inflammation among 16,301 participants in the Third National Health and Nutrition Examination Survey (NHANES III). Measures of kidney function were estimated glomerular filtration rate (GFR) by using the Modification of Diet in Renal Disease equation. RESULTS: The age-adjusted prevalence of anemia (hemoglobin <12 g/dl) at an estimated GFR of 59-30 ml/min/1.73 m(2) was 17% increasing to 56% for those participants with an estimated GFR <30 ml/min/1.73 m(2), while the age-adjusted prevalence of elevated C-reactive protein (CRP >0.21 mg/dl) and reduced 25(OH)D (<20 ng/ml) was 50 and 30%, respectively. After adjusting for demographics, comorbidities and laboratory results, 25(OH)D <20 ng/ml (OR: 1.17; 95% CI: 1.03-1.32; p = 0.014) and log CRP (OR: 3.63; 95% CI: 2.4-5.48 per unit increase of log CRP; p < 0.0001) were associated with hemoglobin levels <12 g/dl. CONCLUSION: This study provides evidence that lower 25(OH)D and higher CRP levels are independently associated with lower hemoglobin concentrations in kidney disease subjects not requiring dialysis. PMID- 19218792 TI - Ciclosporin does not influence bone marrow-derived cell differentiation to myofibroblasts early after renal ischemia/reperfusion. AB - BACKGROUND: Ischemia/reperfusion injury (IRI) is a risk factor for the development of interstitial fibrosis. Previously we had shown that after renal IRI, bone marrow-derived cells (BMDC) can differentiate to interstitial myofibroblasts. Here we hypothesized that the immunosuppressant ciclosporin A (CsA), known for its profibrotic side effect, promotes myofibroblast differentiation of BMDC in the postischemic kidney. METHODS: Using a model of unilateral renal IRI in rats reconstituted with R26-human placental alkaline phosphatase transgenic bone marrow, CsA was administered in a previously defined critical window for differentiation of BMDC to myofibroblasts. We evaluated fibrotic changes in the kidney and myofibroblast differentiation of BMDC on day 14 after CsA treatment. RESULTS: CsA treatment for 14 days led to increased transforming growth factor-beta transcript levels and collagen III deposition in the postischemic kidney. However, neither the total number of alpha-smooth-muscle actin-positive interstitial myofibroblasts, nor the bone marrow-derived fraction thereof was affected by CsA administration, irrespective of dosage and duration of treatment. CONCLUSIONS: In the critical postischemic window of BMDC differentiation to myofibroblasts, CsA did not promote BMDC differentiation to myofibroblasts, suggesting that, in the clinical setting, CsA is not involved in myofibroblastic differentiation of BMDC. PMID- 19218793 TI - How does higher ultrafiltration within the conventional clinical range impact the volume status of hemodialysis patients? AB - AIMS: The higher ultrafiltration (UF) induces poor outcomes. The impact of higher UF on the volume status was investigated. METHODS: 60 hemodialysis (HD) patients were divided into three groups according to the ratio of total UF to post dialysis body weight (TUF/PDW) (<3%, 3-5%, > or =5%). ANP, the ratio of extracellular water to total body water and excess fluid mass (ExF/PDW) by bioimpedance spectroscopy, inferior vena cava diameter by ultrasound were measured at the end of HD. The ratio of post-HD blood volume to pre-HD (BVpost/BVpre) and standardized filtration coefficients (Lpst) of the microvasculature in the vicinity of PDW were calculated. RESULTS: Only Lpst and BVpost/BVpre showed significant differences among the three groups. A stepwise multiple linear regression model revealed that BVpost/BVpre was correlated with TUF/PDW, ExF/PDW and Lpst (R = 0.778, p < 0.001), independently. CONCLUSION: Higher UF causes decreases in BVpost/BVpre and Lpst. BVpost/BVpre was determined by TUF/PDW, ExF/PDW and Lpst. PMID- 19218794 TI - The role of endothelial progenitor cells in acute kidney injury. AB - Acute kidney injury (AKI) is characterized by a sudden impairment of kidney function, which results in the retention of urea and other nitrogenous waste products and in the perturbation of extracellular fluid volume as well as electrolyte and acid-base homeostasis. The dysfunction and apoptosis of tubular epithelial cells are of key importance for the pathophysiological consequences of AKI. However, a growing body of evidence supports the contribution of altered renal vascular structure and function in potentially initiating and extending the initial tubular injury. Vascular injury and dysfunction result in alterations of renal oxygenation and hemodynamics that may have long-term effects in regards to renal function, predisposing to chronic kidney disease. There is growing evidence that endothelial progenitor cells (EPCs) may improve vascular regeneration in different ischemic organs, and recent data suggest that EPCs are mobilized after acute renal ischemia and recruited in ischemic kidney areas and can ameliorate AKI through both paracrine effects and repair of injured microvasculature. The loss of endothelial cell function may represent an important therapeutic target, in which EPCs may show potential importance in ameliorating the acute and chronic effects of ischemic AKI. PMID- 19218795 TI - Sex difference in the prevalence of deep-vein thrombosis in Japanese patients with acute intracerebral hemorrhage. AB - BACKGROUND: Stroke patients often develop deep-vein thrombosis (DVT), a potential cause of pulmonary thromboembolism. Little information is available on DVT in Asian patients with intracerebral hemorrhage (ICH). METHODS: We prospectively enrolled consecutive acute ICH patients. The main exclusion criteria were neurosurgical treatment, early death and coagulation disorders. DVT was evaluated using venous duplex ultrasonography on the day of admission, as well as 7 and 14 days later. Underlying characteristics, stroke features and laboratory data on admission were compared between patients who developed DVT by 14 days and those who did not. RESULTS: A total of 81 (50 men, mean age 65 years, median NIH Stroke Scale, NIHSS, score 12) of 117 Japanese ICH patients were enrolled. DVT was detected in 4 patients on admission and was newly detected in 9 at 7 days. By 14 days, 17 patients (21%) were diagnosed as having DVT without thromboembolic complications, although 1 patient developed pulmonary thromboembolism. DVT was detected in the soleal veins of all 17 patients, followed by the peroneal veins (7 patients). After adjustment for age and related confounders, female sex was the only independent predictor for DVT (odds ratio 6.89, 95% confidence interval, CI, 1.56-36.34, p = 0.014). Female patients with an initial NIHSS score > or =12 had 19 times the risk for DVT compared to men with an NIHSS score <12 (95% CI 2.61-213.77, p = 0.007). CONCLUSIONS: DVT formation was not rare in Japanese ICH patients. Contrary to previous findings reported from western countries, female sex was strongly associated with DVT formation. PMID- 19218796 TI - Deep venous thrombosis after intracerebral hemorrhage, gender and ethnicity: a challenge for therapeutic approaches. PMID- 19218797 TI - Fissured fibrous cap of vulnerable carotid plaques and symptomaticity: are they correlated? Preliminary results by using multi-detector-row CT angiography. AB - PURPOSE: Carotid artery plaque with a disrupted fibrous cap is characterized by a higher tendency to rupture, resulting in a higher rate of transitory ischemic attack and stroke. The purpose of our study was to evaluate whether there is a statistically significant correlation between the presence of fissured fibrous cap (FFC) (assessed by using multi-detector-row CT angiography (MDCTA)) and ipsilateral symptomaticity. MATERIAL AND METHODS: 147 patients (105 males, 42 females; mean age 63 years, range 37-84) with a stenosis of at least 50% or a plaque alteration at sonography were retrospectively studied, yielding a total of 294 carotid arteries, by using a multi-detector-row CT (MDCT) scanner. A search for detection of FFC and a correlation with previously registered data about patients' symptomaticity by using statistical assessment were performed. Each examination was assessed independently by two readers and interobserver agreement was calculated. RESULTS: Among the 147 patients included in the study group, 15 were excluded because of inadequate quality images. In the 132 remaining patients, for a total of 264 carotids assessed, 30 FFCs were detected by using MDCTA and overall there were 36 symptomatic patients (12 ipsilateral symptomatic patients with FFC). A statistical correlation between the presence of FFC and symptomaticity (p = 0.0032) was found. The kappa value between readers was 0.781. CONCLUSIONS: MDCT may depict FFC and the results of our study suggest that FFC may be used as an indicator for prediction of potential cerebrovascular pathology. The interobserver agreement obtained was good. PMID- 19218798 TI - A comparison of acute and long-term management of stroke patients in Barbados and South London. AB - BACKGROUND: To compare health care utilisation between stroke patients living in a middle-income country with similar patients in a high-income country in terms of the type and amount of health care received following a stroke. METHODS: Data were collected from the population-based South London Stroke Register (SLSR) and the Barbados Register of Strokes (BROS) from January 2001 to December 2004. Differences in management and diagnostic procedures used in the acute phase were adjusted for age, sex, ethnic group, living conditions pre-stroke and socio economic status by multivariable logistic regression. Comparison of subsequent management was made for 3 months and 1 year post-stroke. RESULTS: Patients in BROS were less likely to be admitted to a hospital ward (OR 0.22; 95% CI 0.13 0.37), but the difference for the lower use of brain scans in BROS was smaller (OR 0.62; 95% CI 0.25-1.52). Additional adjustment for stroke severity (Glasgow Coma Score) showed that BROS patients were more likely to have a swallow test on admission (OR 2.95; 95% CI 1.17-7.45). BROS patients were less likely to be in nursing care at 3 months (OR 0.37; 95% CI 0.17-0.81), and less likely to be receiving speech and language therapy at 3 months (OR 0.10; 95% CI 0.03-0.33) and 1 year (OR 0.05; 95% CI 0.00-0.55). CONCLUSIONS: The lower use of hospital admission and nursing care at 3 months suggests that in Barbados, family and friends take greater responsibility for patient care around the time of the stroke and in the medium term thereafter. PMID- 19218800 TI - Atherosclerotic carotid vulnerable plaque and subsequent stroke: a high resolution MRI study. AB - BACKGROUND: High-resolution contrast-enhanced magnetic resonance imaging (CEMRI) has been proven to be an effective tool for the identification of carotid atherosclerotic vulnerable plaque, such as a large lipid core and thin fibrous cap. The aim of this study was to evaluate the relationship between carotid plaque characteristics and the types of stroke in patients who had carotid artery (CA) stenosis > or =50%. METHODS: 102 consecutive subjects (mean age 67.2 +/- 10.2 years; 73 males) who initially had ischemic stroke or asymptomatic CA stenosis from 50 to 100% diagnosed by ultrasound were included in this study. Carotid CEMRI, brain MRI and magnetic resonance angiography were performed to understand the infarct patterns and to exclude intracranial artery stenosis. The modified American Heart Association (AHA) plaque classification was used in our study. RESULTS: Our study demonstrated that 45 patients had CA stroke, and 55 patients had lacunar and asymptomatic lesions. The majority of patients had AHA classification type IV-V and VI which presented as vulnerable plaques. Of 63 patients with mild to moderate stenosis (< or =70%), 44 (69.8%) had type IV-V vulnerable plaques, which was significantly higher than those of patients with severe stenosis (>70%; p < 0.001). In CA stroke, the number of patients with a thin or ruptured fibrous cap was twice that of those with a thick and intact fibrous cap. CONCLUSIONS: CEMRI may have important applications in clinical risk evaluations in CA atherosclerosis. Physicians ought to recognize that different types of stroke should be identified by brain MRI detection before invasive therapies. PMID- 19218799 TI - MRI-defined subcortical ischemic vascular disease: baseline clinical and neuropsychological findings. The LADIS Study. AB - BACKGROUND: Subcortical ischemic vascular disease (SIVD) is a common, but often overlooked cause of vascular cognitive impairment. Diagnostic research criteria for SIVD are based on magnetic resonance imaging (MRI) findings including substantial white matter lesions (WML) and multiple lacunar infarcts. Empirical studies validating these imaging criteria are still few. The purpose of the study was to describe the clinical and cognitive characteristics of the MRI-defined SIVD in a mixed sample of functionally independent elderly subjects with WML. METHODS: The subjects of the Leukoaraiosis and Disability (LADIS) study, aged 65 84 years, underwent comprehensive clinical and neuropsychological examinations, and brain MRI at the baseline assessment. The subjects meeting the SIVD imaging criteria (n = 89) were compared to the other subjects of the sample (n = 524). RESULTS: SIVD was associated with lower education, hypertension and, independently, with obesity. The subjects with SIVD had more often motor impairment, a history of falls, and subtle impairment in activities of daily living, but they did not differ for depressive symptoms. SIVD subjects performed significantly inferiorly in tests of global cognitive function, psychomotor speed, attention and executive functions, verbal fluency, and working memory. CONCLUSION: In this population of nondisabled older adults with WML, SIVD was related to specific clinical and functional characteristics. Neuropsychological features included psychomotor slowing as well as deficits in attention and executive functions. PMID- 19218801 TI - Depressed mood after intracerebral hemorrhage: the FAST trial. AB - BACKGROUND AND PURPOSE: Depression is a frequent and important complication of stroke. Few data exist on the prevalence of depression/depressed mood after intracerebral hemorrhage (ICH) and the relationship between depression/depressed mood and the quality of life (QoL) more generally experienced by survivors of ICH. METHODS: Factor Seven for Acute Hemorrhagic Stroke (FAST) was a randomized, multicenter, double-blind, placebo-controlled trial conducted between May 2005 and February 2007 at 122 sites in 22 countries. All patients were evaluated at day 90 after ICH onset for depressed mood and QoL with the Hamilton Depression Rating Scale (HDRS) and the EuroQoL, respectively. Multivariate stepwise logistic regression was used to develop a predictive model for depressed mood at day 90. Relationships between HDRS and EuroQoL scores at day 90 were evaluated with Spearman correlation coefficients. RESULTS: 657 patients were alive at 3 months after ICH onset; 596 (91%) completed the HDRS. Twenty percent reported an HDRS score >10, indicating at least a minor degree of depressed mood; 6% endorsed symptoms of severely depressed mood. Significant predictors of depressed mood included comorbidities (p = 0.0022), moderate to severe neurological impairment according to the National Institutes of Health Stroke Scale (NIHSS) at day 15 (p = 0.0097), physical disability as measured by the Barthel Index (BI) at day 15 (p = 0.0486), and female gender (p = 0.04), but not hemorrhage severity or a history of depression. Irrespective of the presence of post-ICH disability and impairment, the severity of depressed mood at day 90 was significantly correlated with poor QoL. CONCLUSION: Depressed mood affects approximately 20% of ICH survivors and adversely affects QoL. Physicians should be vigilant to ensure that chronically ill and severely impaired survivors of ICH are adequately monitored and treated for depression after ICH. PMID- 19218802 TI - Country-based reference values and impact of cardiovascular risk factors on carotid intima-media thickness in a French population: the 'Paroi Arterielle et Risque Cardio-Vasculaire' (PARC) Study. AB - AIM: To define reference values for mean common carotid artery intima-media thickness (CCAIMT(mean)) from subjects without conventional cardiovascular risk factors. METHODS: This nationwide study involved cardiologists at 246 centres in France. A total of 5,433 subjects with and without conventional cardiovascular risk factors participated. RESULTS: In subjects without risk factors, CCAIMT(mean) was 0.712 +/- 0.122 mm in men and 0.682 +/- 0.105 mm in women (p < 0.0001). Age explained 27.7% (men) and 33.9% (women) of the variance in CCAIMT(mean). Each 10-year increment in age was associated with a sex-adjusted increase in CCAIMT(mean) of 0.049 mm. In subjects with 1 risk factor, CCAIMT(mean) was 0.765 +/- 0.121 (p < 0.0001 vs. subjects without risk factors). CCAIMT(mean) increased continuously with increasing number of risk factors, irrespective of age group. In multivariable analysis, age, sex and number of cardiovascular risk factors appeared independently associated with CCAIMT(mean). CONCLUSION: CCAIMT(mean) may help to identify the population at intermediate cardiovascular risk. As an integrator of both conventional and genetic cardiovascular risk factors at the individual level, its additive value versus other risk scores must be evaluated, particularly in patients in their fourth and sixth decades. PMID- 19218804 TI - Troponin T concentration 3 days after acute ST-elevation myocardial infarction predicts infarct size and cardiac function at 3 months. AB - OBJECTIVES: The evaluation of infarct size and left ventricular function after acute myocardial infarction is important for predicting the subsequent clinical course. This assessment can be achieved by non-invasive imaging methods, but biochemical assays are frequently used as an alternative. We investigated the ability of a single measurement of cardiac troponin T (cTnT) the third morning after onset of ST-segment elevation myocardial infarction (STEMI) to predict infarct size and left ventricular ejection fraction (LVEF). METHODS: The study population consisted of 103 patients with their first STEMI treated with thrombolysis. Blood samples for determination of cTnT were drawn the third morning after onset of symptoms. Infarct size and LVEF were assessed by magnetic resonance imaging 3 months later. RESULTS: Linear regression analysis showed a strong, significant correlation between third-day cTnT and infarct size at 3 months (r = 0.84, p < 0.0001). A significant but inverse and weaker correlation was obtained between third-day cTnT and LVEF (r = -0.63, p < 0.0001). Adjusting for age, gender, infarct location and medication changed the results marginally. CONCLUSIONS: A single cTnT value the third morning after onset of STEMI predicted the final infarct size in this group of patients with their first STEMI, and might be useful for ruling out impaired LVEF. PMID- 19218803 TI - Risk of symptomatic intracerebral hemorrhage in patients treated with intra arterial thrombolysis. AB - BACKGROUND: In intra-arterial (IA) thrombolysis trials, higher rates of symptomatic intracerebral haemorrhage (sICH) were found than in trials with intravenous (IV) recombinant tissue plasminogen activator (tPA); this observation could have been due to the inclusion of more severely affected patients in IA thrombolysis trials. In the present study, we investigated the rate of sICH in IA and combined IV + IA thrombolysis versus IV thrombolysis after adjusting for differences in clinical and MRI parameters. METHODS: In this multicenter study, we systematically analyzed data from 645 patients with anterior-circulation strokes treated with either IV or IA thrombolysis within 6 h following symptom onset. Thrombolytic regimens included (1) IV tPA treatment (n = 536) and (2) IA treatment with either tPA or urokinase (n = 74) or (3) combined IV + IA treatment with either tPA or urokinase (n = 35). RESULTS: 44 (6.8%) patients developed sICH. sICH patients had significantly higher scores on the National Institutes of Health Stroke Scale (NIHSS) at admission and pretreatment DWI lesions. The sICH risk was 5.2% (n = 28) in IV thrombolysis, which is significantly lower than in IA (12.5%, n = 9) or IV + IA thrombolysis (20%, n = 7). In a binary logistic regression analysis including age, NIHSS score, time to thrombolysis, initial diffusion weighted imaging lesion size, mode of thrombolytic treatment and thrombolytic agent, the mode of thrombolytic treatment remained an independent predictor for sICH. The odds ratio for IA or IV + IA versus IV treatment was 3.466 (1.19-10.01, 95% CI, p < 0.05). CONCLUSION: In this series, IA and IV + IA thrombolysis is associated with an increased sICH risk as compared to IV thrombolysis, and this risk is independent of differences in baseline parameters such as age, initial NIHSS score or pretreatment lesion size. PMID- 19218805 TI - Long-term (up to 18 years) clinical and echocardiographic results of mitral balloon valvuloplasty in 531 consecutive patients and predictors of outcome. AB - OBJECTIVES: The study aim was to assess the long-term results (up to 18 years) of mitral balloon valvuloplasty (MBV) and to identify predictors of restenosis and event-free survival. METHODS: The immediate and long-term results for 531 consecutive patients (mean age 31 +/- 11 years) who underwent successful MBV and were followed up for a mean of 8.5 +/- 4.8 years (range: 1.5-18 years) are reported. RESULTS: The mitral valve area (MVA) increased from 0.92 +/- 0.17 to 1.95 +/- 0.29 cm(2) (p < 0.0001). Restenosis was 31 and 19% in patients with mitral echocardiographic score (MES) < or =8. Actuarial freedom from restenosis at 10, 15 and 18 years was 77 +/- 2, 46 +/- 3 and 18 +/- 4% and 86 +/- 2, 62 +/- 4 and 31 +/- 7% for MES < or =8, respectively (p < 0.001). Event-free survival (death, redo MBV, mitral valve replacement, NYHA class III or IV) at 10, 15 and 18 years was 88 +/- 1, 53 +/- 4, and 21 +/- 5% and 93 +/- 2, 65 +/- 5 and 38 +/- 8% for MES < or =8, respectively (p < 0.001). Multivariable Cox regression analysis identified MES >8 (p < 0.0001) and previous surgery (p = 0.043) as predictors of restenosis, and MES >8 (p < 0.0001) and baseline atrial fibrillation (p = 0.03) as predictors of combined events. CONCLUSION: MBV provides excellent long-term results. The baseline clinical and MES characteristics are predictors of outcome. PMID- 19218806 TI - Prenatal diagnosis of retroperitoneal teratoma: a case report and review of the literature. AB - We report a case of retroperitoneal teratoma diagnosed prenatally by serial sonographic examinations in the third trimester. A 29-year-old woman was referred for sonographic evaluation at 33 weeks' gestation because of a fetal intra abdominal mass. Our initial sonographic image suggested a neuroblastoma. Repeat ultrasound images demonstrated an increase in size of the tumor, while the content of the tumor became predominantly solid with areas of calcification. Teratoma should be considered on detection of any cystic or mixed semisolid mass, especially when calcification is present. The fetus was prenatally diagnosed with retroperitoneal teratoma. After birth at 39 weeks, the tumor was removed and histological analysis revealed an immature retroperitoneal teratoma. Intensive monitoring of the changes in ultrasound images of the tumor should provide ground for a precise antenatal diagnosis. PMID- 19218807 TI - Fetal malformations associated with the use of methylphenobarbital and carbamazepine during pregnancy. Two case reports and review of the literature. AB - The use of antiepileptic drugs (AEDs) during pregnancy has been associated with an increased risk of major and minor fetal malformations. This paper describes 2 infants with malformations born to epileptic mothers who used AEDs throughout pregnancy. In the first case, the AED used for seizure control was methylphenobarbital, while in the second case the patient had been prescribed carbamazepine. We noted major and minor congenital malformations in both infants exposed in utero to these anticonvulsant drugs. Pregnant women still experience poor obstetrical care because they report to tertiary centers at the end of their pregnancy or when in labor, making it difficult to provide proper medical care for both the infant and the mother. PMID- 19218808 TI - Incidence of chromosomal abnormalities in the presence of fetal subcutaneous oedema, such as nuchal oedema, cystic hygroma and non-immune hydrops. AB - INTRODUCTION: The authors investigated the incidence of chromosomal abnormalities in subcutaneous oedema detected in the fetus by intrauterine ultrasonography. MATERIAL AND METHOD: In the 10-year period, intrauterine karyotyping was performed in pregnancies with positive ultrasound findings for subcutaneous oedema, such as nuchal oedema, cystic hygroma and non-immune hydrops. RESULTS: Intrauterine karyotyping in fetal subcutaneous oedema was carried out in 434 cases. The chromosomal investigation was made in nuchal oedema in 374 cases, in 120 patients the chromosomal examination was made in the first trimester because of nuchal translucency, and in 254 cases in the second trimester because of nuchal thickening. Cystic hygroma cases (27 patients), non-immune hydrops cases (20 patients), and combined cases of non-immune hydrops and cystic hygroma (13 patients) were investigated separately. In nuchal oedema, pathological karyotypes were detected in 8.33% in the first trimester and in 5.51% in the second trimester. Chromosomal abnormality was found in 48.15, 20, and 53.8% in cystic hygroma, non-immune hydrops, and combined occurrence of non-immune hydrops and cystic hygroma, respectively. Considering all of the changes accompanied by subcutaneous oedema, 50, 25 and 18.75% of the pathological karyotypes was X monosomy, trisomy 18 and trisomy 21, respectively. DISCUSSION: It was important to distinguish nuchal oedema and cystic hygroma, and in the case of non-immune hydrops, it was also important to discuss cases with or without cystic hygroma separately. During the investigations, cases of non-immune hydrops with or without cystic hygroma were evaluated as separate categories. CONCLUSIONS: The authors emphasize the differentiation of the various types of subcutaneous oedema and the importance of precise information about the risks, provided during genetic counselling. PMID- 19218809 TI - Antenatal intravenous immunoglobulin in chronic immune thrombocytopenic purpura: case report and literature review. AB - Immune thrombocytopenic purpura (ITP) may complicate pregnancy and, uncommonly, may cause severe neonatal thrombocytopenia. However, it is difficult to predict which neonates are at risk of severe thrombocytopenia. Direct fetal sampling is not commonly done, as it poses significant risks to the fetus. Furthermore, appropriate antenatal treatment of neonates is controversial. We describe the case of a 32-year-old woman with chronic severe ITP and a previous severely affected infant, pregnant with trichorionic triplets, who was successfully managed with the use of weekly intravenous immunoglobulin 1 g/kg without recourse to direct fetal sampling. PMID- 19218810 TI - Detection of antigens reactive to IgE and IgA during wheat seed maturation and in different wheat cultivars. AB - BACKGROUND: Dietary intake of wheat causes hypersensitivity reactions in patients suffering from IgE-mediated food allergy and coeliac disease. AIM: To study the expression of IgE- and IgA-reactive antigens during wheat seed maturation and in different wheat cultivars. METHODS: Summer wheat was grown in a glasshouse and seeds were harvested at defined maturation stages. Mature seeds were obtained from 13 different defined cultivars. Protein extracts were prepared from different maturation stages and cultivars with a standardized procedure based on seed weight and analysed by IgE and IgA immunoblotting using sera from clinically defined patients suffering from wheat allergy or coeliac disease. RESULTS: With a few exceptions the expression of IgE- and IgA-reactive wheat antigens increased during wheat seed maturation. Wheat cultivars could be identified in which the expression of certain IgE- and IgA-reactive components was strongly reduced or not detectable. CONCLUSIONS: The expression of IgE- and IgA-reactive antigens depends on wheat seed maturation and varies in different wheat cultivars. PMID- 19218811 TI - Evaluation of FITC-induced atopic dermatitis-like disease in NC/Nga mice and BALB/c mice using computer-assisted stereological toolbox, a computer-aided morphometric system. AB - BACKGROUND: The NC/Nga mouse spontaneously develops eczematous atopic dermatitis (AD)-like skin lesions when maintained under conventional conditions, but not when kept under specific pathogen-free (SPF) conditions. Hence, there is a need for an AD model in mice housed under SPF conditions, as this is mandatory for research animals in many countries. METHODS: We evaluated the use of the hapten FITC as an inducer of AD-like disease in NC/Nga and BALB/c mice maintained under SPF conditions. Mice were either untreated or treated with tacrolimus or betamethasone. Using the software Computer Assisted Stereological Toolbox as a stereological method, the mice were sensitized to FITC and the histological efficiency of disease induction with regard to inflammation and CD4+ and CD8+ lymphocytes, in addition to mast cells, was evaluated. The method was validated by comparison to a conventional semiquantitative observer-dependent method. RESULTS: Our findings prove that FITC does indeed induce AD-like lesions in NC/Nga mice with regard to the histological appearance of the mice. However, when evaluating the immunological response in the affected areas of the mice with regard to the CD4/CD8 ratio and the effect of treatment, we found that the immune response in the NC/Nga mice differed from AD skin lesions in humans in certain aspects. CONCLUSIONS: These results emphasize the importance of an assessment of not only the histological but also the immunological appearance of the skin when evaluating AD-like disease in mice as a model for AD in humans. PMID- 19218812 TI - Comparison of acute inflammatory and chronic structural asthma-like responses between C57BL/6 and BALB/c mice. AB - BACKGROUND: The interactions between airway responsiveness, structural remodelling and inflammation in allergic asthma remain poorly understood. Prolonged challenge with inhaled allergen is necessary to replicate many of the features of airway wall remodelling in mice. In both mice and humans, genetic differences can have a profound influence on allergy, inflammation, airway responsiveness and structural changes. METHODS: The aim of this study was to provide a comparative analysis of allergen-induced airway changes in sensitized BALB/c and C57BL/6 mice that were exposed to inhaled allergen for 2 ('acute'), 6 or 9 weeks ('chronic'). Inflammation, remodelling and responsiveness were analyzed. RESULTS: Both strains developed a Th-2-driven airway inflammation with allergen-specific IgE, airway eosinophilia and goblet cell hyperplasia upon 2 weeks of allergen inhalation. This was accompanied by a significant increase in airway smooth muscle mass and hyperresponsiveness in BALB/c but not in C57BL/6 mice. However, airway eosinophilia was more pronounced in the C57BL/6 strain. Chronic allergen exposure (6 or 9 weeks) resulted in an increase in airway smooth muscle mass as well as subepithelial collagen and fibronectin deposition in both strains. The emergence of these structural changes paralleled the disappearance of inflammation in both C57BL/6 and BALB/c mice and loss of hyperresponsiveness in the BALB/c strain. TGF-beta(1 )was accordingly elevated in both strains. CONCLUSION: Airway inflammation, remodelling and hyperresponsiveness are closely intertwined processes. Genetic background influences several aspects of the acute allergic phenotype. Chronic allergen exposure induces a marked airway remodelling that parallels a decreased inflammation, which was largely comparable between the two strains. PMID- 19218813 TI - Promoter polymorphism in the MS4A2 gene and asthma in the Indian population. AB - BACKGROUND: A previous study from our laboratory identified 3-locus risk and protective haplotypes of the MS4A2 gene, a prime candidate for asthma and atopy, that were associated with differential histamine release profiles from basophils and FcepsilonRIbeta transcript levels in the Indian population. METHODS: To explore the role of promoter polymorphisms in the observed association, 4 additional promoter polymorphisms (-752C/T, -654C/T, -426T/C and -109T/C) were investigated in 240 nuclear families with atopic asthma, 237 atopic asthmatics and 221 unrelated controls, all of whom were clinically well characterized. The beta-subunit transcript levels were measured for 15 individuals and were correlated with the associated promoter polymorphisms. RESULTS: We observed a significant association of the -752C/T and -109T/C polymorphisms with asthma, in addition to the already reported association for the INT2 G/A, intron 5 (CA)(n) and 3'-UTR C/T polymorphisms. Additionally, the allele T of the -109T/C polymorphism was associated with a reduced risk of asthma and lower percent peripheral blood eosinophils, thereby pointing towards a protective role for this allele in asthma. Further, the 7-locus haplotypes C_C_T_C_A_16_C and T_C_T_T_G_18_T were identified as the major risk/susceptibility and protective haplotypes, respectively (p < 0.05). Three-locus sliding-window haplotype analysis also identified the -426T/C, -109T/C and INT2 G/A polymorphisms to be in regions of high priority (p < 0.00001). Indeed, the -109T allele was found to be associated with reduced expression levels for FcepsilonRIbeta. CONCLUSIONS: A promoter-dependent mechanism with altered transcriptional regulation of FcepsilonRIbeta may be involved for its association with asthma. These results, therefore, could be useful in predicting the genetic susceptibility of individuals for developing asthma. PMID- 19218814 TI - Probiotic Escherichia coli Nissle 1917 suppresses allergen-induced Th2 responses in the airways. AB - BACKGROUND: Recent clinical trials, epidemiological studies and animal experiments have suggested that probiotics may help suppress the development of allergic responses. OBJECTIVE: To investigate whether the application of the probiotic Escherichia coli strain Nissle 1917 (EcN) protects mice from developing ovalbumin (OVA)-specific T helper-2 responses in the airways. METHODS: OVA specific Th2 responses were induced by 2 intraperitoneal (i.p.) injections with OVA/alum followed by 1 intranasal (i.n.) challenge with OVA. EcN was given orally during the entire sensitization and challenge period, together with OVA/alum during the i.p. sensitizations, or i.n. before or during the airway challenge with OVA. RESULTS: We found that when the bacteria were given together with OVA/alum airway eosinophilia, airway hyper-reactivity, goblet cell metaplasia and IL-5 levels in the bronchoalveolar lavage and mediastinal lymph node cell cultures were reduced. This effect was associated with increased numbers of IFN gamma producing T helper-1 cells and IFN-gamma levels in the airways and strongly increased OVA-specific IgG(2a) titers in the serum. The suppressive effect on airway eosinophilia was dependent on IFN-gamma but not TLR-4. Applying EcN i.n. or orally did not reduce the development of allergen-specific Th2 responses. CONCLUSIONS: Our results suggest that EcN can inhibit the development of allergic responses when the bacteria are present at the site of Th2 cell priming and that this immunomodulatory effect is due to a shift from Th2 to Th1 response. The data support the hypothesis that probiotics may help reduce allergic responses and that EcN may also be used as adjuvant therapy to induce allergen-specific Th1 responses. PMID- 19218815 TI - Natural self-assembly of allergen-S-layer fusion proteins is no prerequisite for reduced allergenicity and T cell stimulatory capacity. AB - BACKGROUND: Recombinant allergen-S-layer fusion proteins display a strongly reduced IgE-binding activity and promote the induction of allergen-specific Th0/1 cells and regulatory T cells. Such fusion proteins show a natural capacity to self-assemble into mono- or double-layer sheets reaching particle-like dimensions of 0.5-2 microm. We were interested in finding out whether self-assembly was crucial for the immunological characteristics of allergen-S-layer fusion proteins. METHODS: The IgE-binding and mediator-releasing capacities of nonassembled and self-assembled rSbpA-Bet v 1, consisting of the major birch pollen allergen Bet v 1 and the S-layer protein SbpA, were compared in inhibition ELISA and basophil activation assays using sera from patients allergic to birch pollen. T cell stimulation was evaluated using Bet v 1-specific T cell clones reactive to distinct epitopes of Bet v 1. Autologous B lymphocytes, monocytes and monocyte-derived dendritic cells were employed to evaluate potential differences in uptake and processing by different antigen-presenting cells. RESULTS: Both rSbpA-Bet v 1 variants showed significantly less IgE-binding and mediator releasing activity than Bet v 1. However, self-assembly further minimized the reduced allergenicity of nonassembled rSbpA-Bet v 1. Both rSbpA-Bet v 1 variants induced comparable proliferation in Bet v 1-specific T cell clones. B cells inappropriately presented either variant of rSbpA-Bet v 1. Self-assembly amplified the T cell stimulatory capacity of monocytes and dendritic cells. CONCLUSIONS: The promising characteristics of allergen-S-layer fusion proteins regarding their potential use for allergy treatment do not depend on the formation of particle-like structures. PMID- 19218816 TI - Re-treatment with omalizumab at one year interval for Japanese cedar pollen induced seasonal allergic rhinitis is effective and well tolerated. AB - BACKGROUND: Seasonal allergic rhinitis (SAR) induced by Japanese cedar pollens is a serious problem in Japan. Omalizumab, a humanized monoclonal anti-IgE antibody, improves symptoms associated with SAR, but a study comprehensively investigating the clinical efficacy, safety and pharmacological effects of omalizumab re treatment has not yet been conducted. METHODS: The open-label, 12-week study was carried out in 34 patients who had been treated with omalizumab in the core study conducted in the previous Japanese cedar pollen season. The study plan including study period, efficacy and safety endpoints, as well as dose regimen, was designed to be the same as in the core study. Omalizumab was administered subcutaneously every 2 or 4 weeks based on the serum IgE level and body weight of each patient. RESULTS: Time course changes in daily nasal symptom medication scores as well as in daily ocular symptom medication scores throughout the pollen period were comparable with those in the omalizumab group in the core study. Serum free IgE levels decreased to below the target level in all patients and were equal to those in the omalizumab group in the core study. The adverse reaction profiles were similar to those in the core study. In addition, the overall incidence of drug-related adverse events and injection site reactions in the re-treatment study did not increase compared with those in the omalizumab group in the core study. There were no serious adverse events, and no anti omalizumab antibodies were detected. CONCLUSION: Omalizumab was effective and safe when consecutively readministered in the second Japanese cedar pollen season. PMID- 19218817 TI - Allergy diagnostic testing in clindamycin-induced skin reactions. AB - BACKGROUND: In clinical practice, clindamycin is increasingly used because of its good tolerability and high efficacy with excellent tissue penetration. However, with increased application of clindamycin, the frequency of side effects such as skin eruptions rises and the need for diagnostic testing to identify clindamycin allergy increases. The aim of this retrospective analysis was to demonstrate the results of skin and challenge tests in cases of clinically suspected clindamycin allergy. METHODS: We evaluated 33 patients with a history of a skin reaction in temporal relation to treatment with clindamycin using standardized patch and prick skin testing. In the case of negative skin tests, oral challenges were performed. RESULTS: Clindamycin hypersensitivity was excluded in 20 patients by negative skin tests and subsequently tolerated oral challenge tests. In 5 patients, positive skin tests strongly suggested delayed-type non-IgE-mediated allergic clindamycin hypersensitivity. In 6 skin test-negative patients (2 patients refused challenge tests), a rash was provoked by controlled challenge tests. CONCLUSIONS: The evaluation of patients with clindamycin-associated skin reactions should include appropriate allergologic tests establishing or excluding the diagnosis of clindamycin hypersensitivity. Combined testing, i.e. skin tests and subsequent challenge tests, appears to be necessary to definitely confirm or rule out the presence of allergic clindamycin hypersensitivity. PMID- 19218818 TI - Allergic airway inflammation initiates long-term vascular remodeling of the pulmonary circulation. AB - BACKGROUND: Asthma and allergic airway inflammation are associated with persistent structural alterations in the bronchi, i.e. airway remodeling. Previous studies have shown that during allergic airway inflammation, similar structural alterations may also be evoked in the pulmonary circulation. However, it remained unknown whether remodeling of the pulmonary circulation is as persistent as airway remodeling. The aim of this study is to investigate the reversibility and resolution of vascular remodeling, induced by allergic airway inflammation. METHODS: A validated mouse model of allergic airway inflammation, utilizing ovalbumin as allergen, was employed. Animals were sacrificed 1 day, 1 week or 1 month after the last allergen exposure, and different parameters of remodeling (smooth muscle mass, proliferation of smooth muscle cells and endothelial cells as well as number of myofibroblasts and procollagen-I-producing cells) were investigated and quantified histologically. RESULTS: Allergen exposure resulted in allergic airway inflammation characterized by a transient leukocyte infiltration and in structural alterations in both airway and vascular compartments. The increase in vascular smooth muscle mass and endothelial proliferation persisted at 1 month after the last allergen exposure. The other parameters and cellular inflammatory response returned to baseline within 1 month after the last allergen challenge. CONCLUSIONS: Based on the findings in this study, we conclude that acute allergic airway inflammation, although being initiated from the airways, is able to evoke similar long-term structural alterations in pulmonary vessels as described for bronchi. PMID- 19218819 TI - Association of apolipoproteins with symptoms of asthma and atopy among schoolchildren. AB - BACKGROUND: There is evidence for an association between serum cholesterol concentrations and asthma, but little is known about the underlying mechanisms. We explored the associations between serum apolipoprotein concentrations and symptoms of asthma and atopy. METHODS: In a population-based cross-sectional study among 10-year-old schoolchildren (n = 462), plasma concentrations of apolipoprotein AI (apoAI) and apolipoprotein B (apoB) were measured. Information on disease symptoms and diagnoses was collected by parental questionnaire. Multivariate logistic regression models were used to analyse the data. RESULTS: High plasma apoAI concentrations (>or=1.74 g/l) were associated with high prevalence of wheeze (OR 3.65; 95% CI 1.43-9.33), and a trend was seen with asthma (OR 3.35; 95% CI 0.94-11.93). In linear analyses, plasma apoAI concentrations were positively related to wheeze (beta-coefficient = 3.20; p = 0.045) and non-atopic wheeze (beta-coefficient = 4.47; p = 0.036), and a trend was found for asthma (beta-coefficient = 3.29; p = 0.099). Overall, plasma apoB concentrations were not associated with either symptoms of asthma or allergy. CONCLUSION: Our results support the hypothesis that high apoAI is associated with the manifestation of asthma and atopy. The underlying pathomechanism still remains to be fully elucidated, but suggests that immune mechanisms may play a central role. However, our results are based on a small study sample and larger studies are warranted to confirm these observations. PMID- 19218820 TI - Efficacy of intravenous gammaglobulin for immunoglobulin G subclass and/or antibody deficiency in adults. AB - BACKGROUND: The clinical significance and efficacy of treating patients who have immunoglobulin (Ig) G subclass deficiency and/or antibody deficiency with Ig replacement therapy has been debated. There are no clear guidelines to recommend intravenous gammaglobulin (IgIV) in these patients as there are few published studies documenting its efficacy. METHODS: We studied in an open-label protocol 10 adult patients with recurrent respiratory infections and IgG subclass and/or antibody deficiency. All patients received monthly IgIV for 12 months and then were observed for 3 months without IgIV infusions. We studied quality of life, incidence of infections, need for antibiotics, frequency of hospitalizations due to infections, IgG subclass and antibody (tetanus and pneumococcal) levels. Innate immunity was evaluated by studying the status of Toll-like receptors and polymorphisms, mannan-binding lectin levels and genotypes. Correction of the immune defects during IgIV therapy was evaluated. RESULTS: Monthly IgIV significantly improved quality of life, decreased the number of infections and the need for antibiotics, and improved IgG subclass and antibody serum levels. No consistent finding of innate immunity could be detected. CONCLUSIONS: IgIV could be beneficial in patients with IgG subclass or antibody deficiency. PMID- 19218821 TI - Cysteinyl leukotrienes induce monocyte chemoattractant protein-1 in human monocyte/macrophages via mitogen-activated protein kinase and nuclear factor kappaB pathways. AB - BACKGROUND: We have previously demonstrated that cysteinyl leukotriene (CysLT) induced monocyte chemoattractant protein-1 (MCP-1) production in monocytes/macrophages. The intracellular signal transduction pathway of MCP-1 production induced by CysLT in human monocytes/macrophages is unclear. METHODS: The activation of mitogen-activated protein kinase (MAPK), including extracellular signal-regulated kinases 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK) and p38 MAPK by phosphorylation, and nuclear factor-kappaB (NF-kappaB) by leukotriene (LT) D(4) and LTC(4) was determined in THP-1 cells, a human monocytic leukemia cell line, and peripheral blood CD14+ monocytes/macrophages. We examined the inhibitory effects of inhibitors of ERK1/2, JNK, p38 MAPK and NF-kappaB and pranlukast as a CysLT1 receptor antagonist on induction of MCP-1 production by LTD(4) and LTC(4). RESULTS: LTD(4) and LTC(4) induced significant phosphorylations of ERK1/2 and JNK, but not p38 MAPK, in THP-1 cells and peripheral blood CD14+ monocytes/macrophages. Pretreatment with the ERK1/2 inhibitor PD98059 and JNK inhibitor SP600125 attenuated MCP-1 production by CysLTs. NF-kappaB activation was induced by addition of LTD(4) and LTC(4). Pretreatment with the NF-kappaB inhibitors caffeic acid phenylethyl ester and MG 132 inhibited MCP-1 production by CysLTs. Pranlukast inhibited phosphorylation of ERK1/2 and JNK, NF-kappaB activation, and the MCP-1 production induced by CysLTs. CONCLUSION: CysLTs induce MCP-1 and this induction is mediated by ERK1/2 and JNK in MAPK, and NF-kappaB pathways via the CysLT1 receptor, for the most part, in human monocytes/macrophages. PMID- 19218822 TI - Selective immunoglobulin M deficiency presenting with recurrent impetigo: a case report and review of the literature. AB - Selective immunoglobulin M (IgM) deficiency is a rare disorder defined by a decreased serum level of IgM and normal levels of other immunoglobulin classes. The disease has not been well described and the cause remains unknown. Patients with IgM deficiency may present with a wide spectrum of clinical manifestations, from asymptomatic to life-threatening infections, including recurrent respiratory and gastrointestinal infections, allergy and autoimmunity. Here, we report a 6.5 year-old otherwise healthy boy with selective IgM deficiency who presented with multiple recurrent impetigo. We reviewed the published data regarding selective deficiency of IgM. PMID- 19218823 TI - Use of oral erythromycin for the treatment of gastrointestinal dysmotility in preterm infants. AB - Milk intolerance due to functional gastrointestinal (GI) dysmotility is a common problem in preterm infants. In the past decade, erythromycin has been used for its motilinomimetric effect to facilitate enteral feeding in preterm infants. Although earlier studies suggested that erythromycin is an effective prokinetic agent, recent randomized control trials (RCTs) reveal conflicting findings. This review assesses the evidence from all RCTs performed to date on erythromycin for preterm infants. The results suggest that oral erythromycin administered in intermediate or high doses as a rescue treatment is associated with a shorter time to attain full enteral feeding and decrease in the duration of requirement for parenteral nutrition. More importantly, the outcome study further indicates that oral erythromycin can reduce the incidence of parenteral nutrition associated cholestasis by almost 50% and decreases the incidence of recurrent septicemia. None of the RCTs reported any sinister adverse effects, in particular, hypertrophic infantile pyloric stenosis or fatal cardiac arrhythmia. Nonetheless, as long-term outcomes have not been fully evaluated, neonatologists should use this treatment cautiously and selectively in preterm infants with moderately severe GI dysmotility. PMID- 19218824 TI - Nimustine (ACNU) plus teniposide (VM26) in recurrent glioblastoma. AB - BACKGROUND: In a previous trial (NOA-01), the combination of nimustine and teniposide showed efficacy in previously untreated glioblastoma (GBM). After establishing temozolomide as standard first-line therapy in GBM patients, the nimustine (ACNU)/teniposide (VM-26) combination has been employed as salvage chemotherapy for recurrent GBM. However, data on the toxicity and efficacy of this regimen in recurrent GBM are lacking. PATIENTS AND METHODS: In two neurooncological centers, all patients with recurrent GBM treated with nimustine (90 mg/m(2), day 1/42) and teniposide (45-70 mg/m(2), days 1-3/42) were analyzed retrospectively for progression-free survival (PFS), overall survival (OS) and toxicity. RESULTS: Thirty-five patients (median age 51 years, range 25-71 years) were identified. Six months after chemotherapy initiation, PFS was 29% and the median OS 6 months; 23% of patients were alive > or = 1 year after initiation of nimustine-teniposide chemotherapy. Grade 4 hematotoxicity was observed in 12 of 35 patients (34%) and in 14 of 83 evaluable chemotherapy courses (17%). CONCLUSIONS: The benefit of the nimustine-teniposide combination is moderate in patients with recurrent GBM. The data support the efficacy of the nimustine teniposide chemotherapy, but the rate of high-grade hematotoxicity is increased. PMID- 19218825 TI - Stereotactic body radiation therapy using three fractions for isolated lung recurrence from colorectal cancer. AB - OBJECTIVE: To determine the feasibility of stereotactic body radiotherapy (SBRT) using 3 fractions for isolated colorectal lung metastases. METHODS: From June 2003 to December 2006, 13 cases of isolated pulmonary metastasis from colorectal cancer were treated by SBRT due to an inoperable state (7 patients), or the patient's refusal to undergo surgical excision (6 patients). All patients underwent chemotherapy for salvage treatment. SBRT doses ranged from 39 to 51 Gy in 3 fractions. Nine patients had a solitary lesion, 3 patients had 2 lesions, and 1 patient had 3 lesions. Median tumor volume for the 18 lesions was 5.9 ml (range 1.6-45 ml). RESULTS: Follow-up duration was 15-57 months. Three-year overall survival, local control and progression-free survival rates were 64.7, 52.7 and 11.5%, respectively.Univariate analysis showed that total internal target volume was a significant prognostic factor for local control. During the follow-up, 11 of the 13 patients experienced local recurrence, distant metastasis or both. The most frequent site of failure was in a nontargeted lung region. No severe complication was attributed to SBRT. CONCLUSION: Our study suggests the potential feasibility of SBRT for selected patients with 1-3 small metastatic nodules. A further larger-scale study is required to define the indications for SBRT in cases with isolated pulmonary metastasis from colorectal cancer. PMID- 19218826 TI - Cranial prophylactic irradiation in locally advanced non-small cell lung carcinoma: current status and future perspectives. AB - As a result of improved local and regional control with aggressive multimodality protocols, the brain has become one of the major sites of relapse in patients with locally advanced non-small cell lung carcinoma (LA-NSCLC). The demonstrated efficacy of prophylactic cranial irradiation (PCI) in small-cell lung carcinoma led to studies of its effectiveness in LA-NSCLC, which indicated that PCI also has a high potential to reduce the incidence or delay the occurrence of brain metastases in this patient group. This report provides an extensive review of the current evidence from nonrandomized and randomized trials regarding the use of PCI in LA-NSCLC and discusses related key issues including risk factors, patient selection criteria, timing of PCI, preferred PCI dosing scheme, toxicity profile and potential novel PCI techniques. PMID- 19218827 TI - Reliability and validity of the Massachusetts general hospital cognitive and physical functioning questionnaire. AB - BACKGROUND: We have recently developed the Massachusetts General Hospital Cognitive and Physical Functioning Questionnaire (CPFQ), a brief scale to measure cognitive and executive dysfunction in mood and anxiety disorders, and we here report on its reliability and validity. METHODS: The internal consistency of the CPFQ was assessed by computing Cronbach's coefficient alpha based upon the average intercorrelation of the 7 items of the CPFQ in a sample of depressed outpatients and by factor analyzing data from the same sample to confirm that the scale is unifactorial and measuring a single construct. Test-retest reliability of the CPFQ was assessed in a different sample of depressed outpatients by computing Pearson's correlation coefficient between pretreatment screening scores and pretreatment baseline scores. Sensitivity to change of the CPFQ was assessed by computing the dependent t test for the subjects in the active treatment condition in the second sample of depressed outpatients. Finally, convergent validity for the CPFQ was assessed in two different ways. RESULTS: We found that the CPFQ is a unifactorial scale, with strong internal consistency. It has good temporal stability as indicated by high test-retest reliability. The CPFQ was also found to be sensitive to change with treatment and displayed convergent validity by significant correlations with other measures of sleepiness, fatigue, apathy and neuropsychological functioning. Although, as expected, the CPFQ was significantly correlated with a measure of depression, the moderate correlation (r approximately 0.30) indicates that the CPFQ is measuring a different construct. CONCLUSION: In summary, the CPFQ is a unifactorial scale, with strong internal consistency, good temporal stability and sensitivity to change with treatment. Further studies will be needed to assess the validity and reliability of this instrument in other psychiatric and neuropsychiatric conditions associated with cognitive dysfunction. PMID- 19218828 TI - The effects on mood of adjunctive single-family and multi-family group therapy in the treatment of hospitalized patients with major depression. A 15-month follow up study. AB - BACKGROUND: Family-based interventions have been shown to be effective in the treatment of depression, but they have seldom been studied in hospitalized depressed patients. This study assesses the value of the additional use of single family or multi-family group therapy within this patient population. METHODS: Eighty-three patients were randomly assigned to: (1) the treatment programme as usual (n = 23), (2) treatment as usual combined with single-family therapy (n = 25) or (3) treatment as usual combined with multi-family group therapy (n = 35). Follow-up assessments were made at 3 months and 15 months. RESULTS: Multi-family group and single-family therapy conditions showed significantly higher rates of treatment responders than the group receiving the usual treatment (49, 24 and 9%, respectively), and higher rates of patients no longer using antidepressant medication (26, 16 and 0%, respectively) at 15 months. Partners taking part in the family treatments were significantly more likely to notice the improvements in the emotional health of the patient early on compared to those in the treatment as usual condition. CONCLUSIONS: This study suggests that single-family and multi-family therapy may benefit hospitalized patients with major depression, and may help the partners of the patients to become aware of the patient's improvement more quickly. PMID- 19218829 TI - Worry exposure versus applied relaxation in the treatment of generalized anxiety disorder. AB - BACKGROUND: Worry exposure (WE) is a core element of cognitive-behavioral treatment for generalized anxiety disorder (GAD). Its efficacy as a stand-alone treatment method (without further cognitive-behavioral therapy interventions) has never been tested.We aimed to examine whether WE alone is as efficacious as the empirically supported stand-alone treatment for GAD, applied relaxation (AR). METHODS: In a randomized controlled study, 73 outpatients meeting DSM-IV criteria for GAD as primary diagnosis were allocated to either WE or AR or a waiting list control group; in a 2nd randomization procedure the waiting list subjects were reallocated to WE or AR. The treatment was manualized (15 sessions with WE or AR), included 6-month and 1-year follow-ups, as well as last observation carried forward and completer analyses, and was controlled for allegiance effects.The Hamilton Anxiety Rating Scale and the State-Trait Anxiety Scale were used as primary outcome measures. Self-report scales of anxiety, worrying and depression including negative metacognition about worrying and thought suppression served as secondary outcome measures. RESULTS: The dropout rate was moderate. The pre /posttreatment effects were high for the Hamilton Anxiety Rating Scale (standardized mean difference >1) and for the State-Trait Anxiety Inventory (standardized mean difference >0.87). The proportion of patients reaching high end state functioning was 48% (WE) and 56% (AR). WE and AR did not differ with regard to dropout rate or treatment effects. The treatment effects were stable at 6 month and 1 year follow-up. CONCLUSION: This is the first study to show that a stand-alone exposure in sensu technique--WE--is efficacious in the treatment of GAD. Both AR and WE seem to represent effective principles of change in GAD. PMID- 19218830 TI - Sensitivity to change and predictive validity of the MOODS-SR questionnaire, last month version. AB - BACKGROUND: Instruments that are intended to measure change over time need to emphasize sensitivity to change as a central property. The aims of this report are to test whether the MOODS-SR, a measure of mood spectrum symptomatology, is sensitive to changes during acute and continuation treatment of depression and whether residual mood spectrum symptoms predict relapse in the subsequent 6 months. METHODS: The study sample includes 316 patients with nonpsychotic depression participating in the protocol 'Depression: the search for treatment relevant phenotypes'. Patients were initially randomized to selective serotonin reuptake inhibitors or interpersonal psychotherapy and then treated for 9 months using an algorithm-based protocol. Measures of mood symptomatology included the self-report version of the structured clinical interview for mood spectrum (MOODS SR), the Quick Inventory for Depressive Symptomatology and the Hamilton Rating Scale for Depression. RESULTS: Repeated-measures ANOVA indicates that during the acute phase MOODS scores decrease significantly from baseline to weeks 6 and 12. This decrease was significantly different (p < 0.001) between those who remitted and those who did not remit on the depressive, the rhythmicity component and the total score. Nonrelapsing subjects had stable scores across the continuation phase, while among relapsing subjects, a significant increase was found in the depressive component (p < 0.001), the rhythmicity component (p = 0.024) and the total score (p < 0.001), at 2 months, followed by a decrease from 2 to 6 months. Scores on the depressive component at the entry into continuation predicted relapse in the subsequent 6 months. CONCLUSIONS: Our findings suggest that the MOODS-SR is sensitive to change in depression status and may help the clinician to detect symptoms and signs not considered by established symptom severity scales. PMID- 19218831 TI - The neuro-ecology of resource localization in Drosophila: behavioral components of perception and search. AB - From the moment an adult fruit fly ecloses, its primary objective in life is to disperse and locate the source of an attractive food odor upon which to feed and reproduce. The evolution of flight has greatly enhanced the success of fruit flies specifically and insects more generally. Control of flight by Drosophila melanogaster is unequivocally visual. Strong optomotor reflexes towards translatory and rotational visual flow stabilize forward flight trajectory, altitude and speed. The steering responses to translatory and rotational flow in particular are mediated by computationally separate neural circuits in the fly's visual system, and gaze-stabilizing body saccades are elicited by threshold integration of expanding visual flow. However, visual information is not alone sufficient to enable a fruit fly to recognize and locate an appropriately smelly object due in part to the relatively poor resolution of its compound eyes. Rather, the animal uses an acute sense of smell to actively track odors during flight. Without a finely adapted olfactory system, the fly's remarkable visual capabilities are for naught. The relative importance of vision is apparent in the cross-modal fusion of the two modalities for stable active odor tracking. Olfactory processing in Drosophila is shaped by ecological and functional forces which are inextricably linked. Thus physiologists seeking the functional determinants of olfactory coding as well as ecologists seeking to understand the mechanisms of speciation do well to consider each others' point of view. Here we synthesize a broad perspective that integrates across ultimate and proximate mechanisms of odor tracking in Drosophila. PMID- 19218832 TI - Chromatin dynamics coupled to DNA repair. AB - In order to protect and preserve the integrity of the genome, eukaryotic cells have developed accurate DNA repair pathways involving a coordinated network of DNA repair and epigenetic factors. The DNA damage response has to proceed in the context of chromatin, a packaged and compact structure that is flexible enough to regulate the accession of the DNA repair machinery to DNA-damaged sites. Chromatin modifications and ATP-remodeling activities are both necessary to ensure efficient DNA repair. Here we review the current progress of research into the importance of chromatin modifications and the ATP-remodeling complex to the DNA damage response, with respect to the sensing and signaling of DNA lesions, DNA repair and the processes that restore chromatin structure. PMID- 19218833 TI - Genome-scale network analysis of imprinted human metabolic genes. AB - System analysis of metabolic network reconstructions can be used to calculate functional states or phenotypes. This provides tools to study the metabolic effects of genetic and epigenetic properties, such as dosage sensitivity. We used the genome-scale reconstruction of human metabolism (Recon 1) to analyze the effect of nine known or predicted imprinted genes on metabolic phenotypes. Simulations of maternal deletion of ATP10A indicated an anabolic metabolism consistent with the known clinical phenotypes of obesity. The abnormal expression of the other genes affected fewer subsections of metabolism consistent with a lack of established clinical phenotypes. We found that four of nine genes had metabolic effect as predicted by the Haig's parental conflict theory. PMID- 19218835 TI - A discussion of two articles on workplace bullying. PMID- 19218836 TI - Defining nursing practice: the ANA Social Policy Statement, 1980-1983. AB - This article examines the initial development of the American Nurses Association's Social Policy Statement (SPS). The methodology used is the Social History of Technology. This approach provided the means to treat the SPS as an information practice or tool that resides within a technological system composed of skills, tools, and knowledge. The data for analysis include archive material and secondary sources. This study concludes that development of the SPS was inherently a local exercise in prioritizing, organizing, and promoting certain skills, tools, and knowledge associated with nursing practice, an exercise that integrated contextual influences with local realities. The study also provides a means to reflect on the development of contemporary information practices. PMID- 19218837 TI - Historical voices of resistance: crossing boundaries to praxis through documentary filmmaking for the public. AB - This article contextualizes my forthcoming study of a particular instance of resistance in nursing history, the Cassandra Radical Feminist Nurses Network, and examines how nursing history can be produced as public media to advance progressive ideas about nurses and transformative and emancipatory nursing and healthcare. It argues that nurse-generated documentary filmmaking is a natural extension of theory and practice, linking several disciplinary and conceptual fields to support a praxis that is situated at the intersection of nursing, critical theory, and the humanities. PMID- 19218838 TI - Expanding the role of women as nurses during the American Civil War. AB - Using the method of social history, this article looks at the many roles filled by women during the American Civil War. Through primary documents such as letters and diaries, the article captures a moment in time when society began to accept an expansion of women's allowed behaviors because of the war. One such field of endeavor was nursing, in which, contrary to Victorian notions, women proved their ability to withstand gruesome conditions in an effort to care for injured and ill soldiers. The purpose of this article is to reconstruct a brief period in which society began to accept role expansion for American women because of the demands of the American Civil War. PMID- 19218839 TI - Margaret Shanks, nurse to Susan B. Anthony: exploring the extraordinary in the "ordinary" nurse. AB - This historical effort explores the life and professional practice of the relatively unknown nurse who attended the well-known suffragist Susan B. Anthony during the last months of her life. It examines nursing practice in the United States at the end of the 19th century through the life and voice of this nurse. The study affirms the value in social history of understanding ordinary lives and practice of nurses. It analyzes the professional, personal, and social interactions between nurse and patient that change each, all within a most significant historical, social, and political context. PMID- 19218840 TI - Reinterpreting the concept of "nursing" in the Korean context. AB - Florence Nightingale acts as the threshold dividing "modern" nursing and "premodern" nursing. The history of modern Korean nursing is not exceptional. It is not surprising that the history of modern Korean nursing describes the advent of "Western" nursing as it was delivered from Western medical professionals. Although traditional Korean medical doctors, Hanuisa, largely practice at traditional Korean health clinics and hospitals, the nurses who work at these traditional Korean health clinics and hospitals are trained at the same nursing education programs as those who work in Western medical clinics and hospitals. The purpose of this study was to find a concept of "nursing" in the context of traditional Korean medicine, which traditional Korean nursing in modern era may inherit and expand upon, by reinterpreting the concept of nursing in terms of a philosophy that supports traditional Korean medicine. This article utilized a philosophical analysis of literature that is essential to the reinterpretation of this concept. In terms of core concepts, nursing and traditional Korean medicine share common features with each other. On the basis of these common features, cowork in practice should be more developed and articulated. This will add new characteristics to the history of Korean nursing. PMID- 19218841 TI - Nurses in the Nazi "euthanasia" program: a critical feminist analysis. AB - From 1940 to 1945, Nazi Germany conducted a program of killing institutionalized psychiatric patients. Known as "euthanasia," this killing program included the administration of lethal doses of medication given largely by nurses. The purposes of this article are to (1) describe the historical context in which nurses' participation in the Nazi euthanasia program occurred; (2) present a recently unsealed narrative testimony of a nurse accused of active participation in the euthanasia program; and (3) analyze this account from a critical-feminist perspective, with a focus on its epistemological salience for contemporary nursing. PMID- 19218842 TI - A method for historicizing lived experience. AB - The purpose of this article is to describe a method used to historicize the lived experience of nurse caring by the culture of white Northern women during the American Civil War. Primary sources included unpublished and published Civil War letters, journals, diaries, memoirs, reminiscences, narratives, and records from 1861 through 1911. The phenomenological description was created using van Manen's approach for hermeneutic phenomenology and Drew's method for examining the researcher's preunderstanding of a phenomenon. The phenomenological description was historicized using Scott's concept of historicizing and Leininger's Sunrise Model as a framework. The method used to historicize the lived experience of American Civil War nurse caring provides a foundation for examining American nurse caring in other time periods in order to reveal the phenomenon of nurse caring during these periods as well as the cultural and social forces that have shaped American Nursing. PMID- 19218843 TI - Catholic sister nurses in Selma, Alabama, 1940-1972. AB - This study analyzes the activities of religious sister nurses as they confronted racism in the American South from 1940 to 1972. Selma was chosen as a case study because, in the 1960s, events in that southern town marked a turning point in the civil rights movement in the United States. This is a story about the workings of gender, race, religion, and nursing. The sisters' work demonstrates how an analysis of race in nursing history is incomplete without an understanding of the roles that a number of Catholic religious women took in reaching out to African Americans in the Deep South. PMID- 19218846 TI - Vestibular pathology presenting as behavioral problems in a child with Down Syndrome: a case report. AB - Here we present a case of superior semicircular canal dehiscence (SSCD) in a non verbal 17-year-old male patient with trisomy 21 that presented with behavioral problems, instead of the more typical symptoms of episodic noise-induced vertigo. SSCD syndrome is characterized by sound-, or pressure-induced vertigo. Patients often complain of vertigo or oscillopsia evoked by loud noises, which may be either external sounds or those elicited by the patients themselves when they are speaking. In cases of SSCD, auditory testing will typically reveal a conductive hearing loss on the affected side while high-resolution temporal bone computed tomography will demonstrate bony dehiscence overlying the ipsilateral superior semicircular canal. Here we present a report of SSCD in a patient with Down Syndrome who was without speech and reacted to noise stimulation with behavioral outbursts. In this case, simple modifications, such as noise avoidance and hearing protection were able to effect positive changes in behavior and improve social interactions. This case underscores the importance of a broad differential diagnosis, and the need to consider alternate presentations, in children with Down Syndrome or any child who is otherwise unable to express himself. PMID- 19218847 TI - Commentary on the use of acupuncture in chronic pediatric pain. AB - The use of acupuncture for pain in pediatrics is a long-standing practice in Eastern cultures. Despite growing interest in the West, there has been relatively little systematic research on acupuncture for chronic pediatric pain. In particular, there is a paucity of randomized clinical trials testing the efficacy of acupuncture for chronic pain problems in pediatric populations. This commentary briefly reviews the history of acupuncture for pain and includes a summary of extant findings regarding potential mechanisms of its analgesic effects. Key areas for future research to advance the application of acupuncture to chronic pediatric pain problems are outlined. PMID- 19218848 TI - Global developmental delay in a 10-month-old infant boy. AB - A 10-month-old boy was seen for the first time for a health supervision visit by a pediatrician. A brief review of the child's medical history did not reveal any specific problems. On physical examination, the pediatrician found an alert, smiling child, but she was surprised by the following observations: unable to sit without support, absent pincer grasp, no audible language, unilateral exotropia, and microcephaly. Expansion of the medical history revealed an uneventful full term prenatal course and normal vaginal delivery. The mother denied use of alcohol or other drugs/medications during the pregnancy. She did not have a recent history of any infections, unexplained fevers, or high risks for sexually transmitted disease. The baby cried spontaneously and the parents reported no resuscitation efforts. There were early feeding problems associated with a poor suck and gastroesophageal reflux. The parents were healthy and this was their first child. Family history was negative for early problems in child development or any neurological conditions. Parents were high school graduates without any learning problems; they were both employed in retail sales with a steady employment history. The pediatrician then took a second look at the child and discovered truncal hypotonia, extremity hypertonia, tongue protrusion, and a broad mouth. She concluded that the child had a global developmental delay, including delays in motor, language, and social development. PMID- 19218849 TI - Basic meta-analysis: conceptualization and computation. PMID- 19218850 TI - Developmental and behavioral disorders grown up: attention deficit hyperactivity disorder. AB - Pat was diagnosed with attention deficit hyperactivity disorder-combined type in the second grade and has been treated primarily with stimulants, albeit on an inconsistent basis. Pat is now a 22-year-old male, who is a second year student at a local community college. He attained a 2.1 GPA during his first year of postsecondary studies, doing quite well in several classes yet rather poorly in several others. Pat has a part-time job at a fast food restaurant, a position that he has held for approximately 6 months. Before his current job, Pat worked as a dishwasher at a local restaurant. He was fired from this position due to chronically being late to work. Pat stated that he gets along well with his peers, although his mother reported that Pat has a history of being able to make friends, "yet not keep them." Pat's mother also reported that Pat has traditionally gravitated toward peers who were "not especially good influences." She described various antisocial acts that Pat was involved with during high school. Pat reported that he began experimenting with alcohol and cigarettes in ninth grade and marijuana shortly thereafter. He stated that he drinks less often than he smokes marijuana ("several times a week"). Pat denied legal difficulties, although his mother reported that Pat was asked to leave the dormitory during his first semester due to an alcohol-related incident. Pat is not currently involved in psychotherapy and is prescribed extended release methylphenidate. He reported that he took his stimulant whenever he had to do a lot of work for school. PMID- 19218851 TI - Healthy Steps: a systematic review of a preventive practice-based model of pediatric care. AB - The preventive role of anticipatory guidance in pediatric practice has gained increasing importance over the last two decades, resulting in the development of competing models of practice-based care. Our goal was to systematically evaluate and summarize the literature pertaining to the Healthy Steps Program for Young Children, a widely cited and utilized preventive model of care and anticipatory guidance, Medline and the bibliographies of review articles for relevant studies were searched using the keywords: Healthy Steps, preventive care, pediatric practice and others. Other sources included references of retrieved publications, review articles, and books; government documents; and Internet sources. Relevant sources were selected on the basis of their empirical evaluation of some component of care (e.g., child outcomes, parent outcomes, quality of care). From 21 identified articles, 13 met the inclusion criteria of empirical evaluation. These evaluations were summarized and compared. Results indicated that the Healthy Steps program has been rigorously evaluated and shown to be effective in preventing negative child and parent outcomes and enhancing positive outcomes. Despite limited information concerning cost effectiveness, the Healthy Steps Program provides clear benefit through early screening, family-centered care, and evidence-based anticipatory guidance. It is recommended that the Healthy Steps program be more widely disseminated to relevant stakeholders, and further enhanced by improved linguistic and cultural sensitivity and long term evaluation of cost effectiveness. PMID- 19218852 TI - Pediatricians' role in providing mental health care for children and adolescents. PMID- 19218854 TI - Role of toll-like receptors 2 and 4, and the receptor for advanced glycation end products in high-mobility group box 1-induced inflammation in vivo. AB - High-mobility group box 1 (HMGB-1) has been reported as a "late" proinflammatory mediator in sepsis. In vitro data have shown that HMGB-1 can induce activation of intracellular signaling pathways via interaction with at least three pattern recognition receptors: Toll-like receptor (TLR) 2, TLR-4, and the receptor for advanced glycation end products (RAGE). The objective of this study was to investigate the role of these receptors in the in vivo response to HMGB-1. Therefore, we first performed a time-series experiment with wild-type (Wt) mice. High-mobility group box 1 induced time-dependent elevations of TNF-alpha, IL-6, monocyte chemoattractant protein 1, and thrombin-antithrombin complex levels in peritoneal lavage fluid and plasma. This inflammatory reaction was accompanied by a prominent and sustained rise in neutrophil counts in the peritoneal cavity. We next administered HMGB-1 to Wt, TLR-2, TLR-4, and RAGE mice. All genotypes showed similar plasma levels of TNF-alpha, IL-6, IL-10, and thrombin-antithrombin complex at 2 h after intraperitoneal injection of HMGB-1. Compared with Wt mice, both TLR-4 and RAGE mice displayed lower TNF-alpha and IL-6 concentrations and lower neutrophil numbers in their peritoneal lavage fluid. In contrast, TLR-2 mice showed increased levels of TNF-alpha and IL-6 in their peritoneal cavity relative to Wt mice. These data indicate that HMGB-1 induces release of cytokines, activation of coagulation, and neutrophil recruitment in vivo via a mechanism that at least in part depends on TLR-4 and RAGE. PMID- 19218855 TI - Retraction. PMID- 19218856 TI - Focused emergency echocardiography: lifesaving tool for a 14-year-old girl suffering out-of-hospital pulseless electrical activity arrest because of cardiac tamponade. AB - Cardiac arrest presenting as pulseless electrical activity (PEA) is associated with poor outcome. Its major underlying causes (e.g. cardiac tamponade, pulmonary embolism, tension pneumothorax or hypovolaemia) are difficult to detect reliably in an out-of-hospital setting. We here present a case of a 14-year-old girl suffering from PEA because of cardiac tamponade 4 weeks after surgical closure of a ventricular septal defect. Immediate focused echocardiography on scene by an emergency physician showed a large pericardial effusion that had led to cardiac tamponade and finally to a PEA cardiac arrest. Immediate pericardiocentesis was carried out. The girl progressed to complete neurological recovery. This case report demonstrates that focused emergency echocardiography may be useful for the diagnosis of pericardial tamponade leading to cardiac arrest and this diagnosis can be made out-of-hospital by an appropriately trained physician. PMID- 19218857 TI - Pediatric mandibular reconstruction after benign tumor ablation using a vascularized fibular flap. AB - This study evaluated the aesthetic and functional outcomes of vascularized fibular flaps used for pediatric mandibular reconstruction. Six pediatric patients ranging in age from 12 to 15 years underwent mandibular reconstruction using a vascularized fibula flap with a skin paddle, after benign tumor ablation. All of the flap transfers were successful, and there were no complications. Panorex radiographs showed good bone union in all patients. The flap conferred adequate support for dental rehabilitation. The mean follow-up was 24.2 months. The aesthetic and functional outcomes were normal. No patient had recurrence. A vascularized fibular flap is a very reliable method for reconstructing the pediatric mandible after benign tumor ablation. PMID- 19218858 TI - Prognostic significance of stromal eosinophilic infiltration in oral squamous cell carcinoma. AB - Tumor-associated tissue eosinophilia (TATE) has been described in many sites, including head and neck. The aim of this study was to investigate the prevalence and possible role of TATE as a prognostic factor in oral squamous cell carcinoma (OSCC). Seventy-six consecutive patients with a diagnosis of OSCC were evaluated, and the number of eosinophils was obtained in 3 different areas in the tumor. The possible role of TATE as a prognostic factor in OSCC was investigated with respect to tumor differentiation; perineural, vascular, and muscular invasion (histopathologic parameters); and locoregional metastasis (clinical parameter). Tumor-associated tissue eosinophilia was low in most cases in 3 different areas. The mean eosinophil count was higher in the invasive front of tumor than intratumoral stroma and stroma subjacent the surface epithelium. The relations between TATE and tumor differentiation; perineural, vascular, and muscular invasion; and locoregional metastasis were not statistically significant (P > 0.05). Tumor-associated tissue eosinophilia has no correlation with prognostic parameters in OSCC. PMID- 19218859 TI - Three-dimensional facial surface analysis of patients with skeletal malocclusion. AB - Three-dimensional (3D) laser surface scanning analysis has taken hold in orthodontics, as well as craniomaxillofacial and plastic surgery as a new tool that can navigate away from the limitations of conventional two-dimensional methods. Various techniques for 3D reconstruction of the face have been used in diagnosis, treatment planning and simulation, and outcomes follow-up. The aim of the current prospective study was to present some technical aspects for the assessment of facial changes after orthodontic and orthognathic surgery treatment using 3D laser surface scanning. The technique proposed for facial surface shape analysis represented three-dimensionally the expected surgical changes, and the reduction of the postoperative swelling was verified. This study provides technical information from the data collection to the 3D virtual soft-tissue analysis that can be useful for diagnostic information, treatment planning, future comparisons of treatment stability or facial postoperative swelling, and soft-tissue profile assessment. PMID- 19218860 TI - Early surgical intervention for Japanese children with infantile hemangioma of the craniofacial region. AB - Although the indications for surgical intervention in patients with infantile hemangioma during the proliferation and early involution are controversial, it is considered in certain cases. Thirty-two Japanese patients (3 boys and 29 girls) were surgically treated at 24 months or earlier to resolve complications (obstruction of the visual axis, ulceration and bleeding, and deviation of the nasal septal cartilage) or to achieve esthetic improvement. Results were evaluated by 3 indicators: outcome, scarring, and resolution of the functional problem. Excellent or good surgical results were obtained in 81.2% of all patients and in 95% of patients in nonorbital areas. Earlier surgical intervention can be an option for treating infantile hemangioma in the craniofacial region. PMID- 19218861 TI - Chorionic villus sampling and hemangiomas. AB - Chorionic villus sampling (CVS) is a widely used and safe method of prenatal diagnosis. In the 1990s, concerns were raised at several medical centers that there was an increased risk to the exposed fetus for the occurrence of limb deficiencies, hemangiomas, and other vascular disruption defects. The risk was greater when the procedure was performed earlier in pregnancy, such as 8 or 9 weeks of gestation. The postulated mechanisms for the fetal injury included blood loss, hypoperfusion, hypoxia, endothelial cell damage, hemorrhage, and tissue loss. The effect was most common in tissues with end arteries, such as digits, tongue, brain stem, and intestine. The associated hemangiomas were infantile hemangiomas. They were more common on the head, neck, and thorax and more often multiple in infants exposed to CVS. One postulated mechanism for the occurrence of these hemangiomas is embolization of angioblasts or endothelial cells from the placenta to the fetal skin. A question to be answered is whether the infantile hemangiomas in children exposed to CVS differ in immunohistologic characteristics from similar hemangiomas in children not exposed to CVS during pregnancy. PMID- 19218863 TI - The 30-year tale of a fellow P&S classmate: a journey along the path of craniosynostosis surgery. PMID- 19218864 TI - Skills of the surgeon, eye of an artist, mind of the analyst. AB - Plastic and reconstructive surgery challenges the surgeon in ways often difficult to comprehend. Not only does the surgeon have to perfect the appropriate techniques for the surgical intervention, but he/she also assesses the patient with the eye of the artist and the mind of a psychoanalyst. Working with children and adolescents and their parents requires even more complex skills because it is required to assume the position of the parent to interpret their wishes, hopes, and fears. Over decades, John Mulliken has been the exemplar of the surgeon able to take on these manifold challenges. Dr Mulliken's special focus on healing the cleft lip and palate has offered to generations of young people and their families a life freer of the complex and sometimes devastating consequences of living with a deformity. Moving for ever earlier intervention, Dr Mulliken broke new ground surgically but from a psychological perspective offered the chance to avoid a host of psychological problems. Dr Mulliken's surgical intervention for nevi and other vascular malformations has had a long-lasting psychological impact on the lives of children. This article traces the place of plastic and surgical reconstructive surgery as a field intertwined with the psychological development of the individual and describes the ways in which Dr Mulliken's approaches have enhanced the well-being of so many. PMID- 19218862 TI - Hemifacial microsomia: from gestation to childhood. AB - Hemifacial microsomia (HFM) is a variable, complex malformation involving asymmetric hypoplasia of the face and ear. Little is known about the risk factors for or consequences of HFM. In this study, we describe 3 studies that have been or are currently being conducted to further our understanding of this malformation. The first completed study examined whether HFM risk is related to maternal exposures that may affect blood flow. In that case-control study, interview data from 230 mothers of children in the case group and 678 mothers of children in the control group suggested that maternal use of vasoactive medications in the first trimester, particularly in combination with cigarette smoking, was associated with increased risks of HFM. The second study is currently underway, in which we are evaluating whether HFM risk is related to genetic variation in pathways associated with vasculogenesis and hemostasis, using DNA collected in the first study. The third ongoing study observes children with HFM to identify psychosocial, cognitive, dental, and medical sequelae. When the children from the original case-control study are 6 or 7 years of age, mothers and teachers complete self-administered questionnaires that cover a wide range of psychosocial development domains. Preliminary analyses of 115 case and 314 control children suggest that children with HFM may have worse teacher reported academic performance and possibly higher levels of internalizing behavior problems than control children. When data on the full study sample are available, further analyses will determine whether the preliminary findings remain and if they vary by HFM phenotype, parenting style, or indicators of social risk. PMID- 19218865 TI - Hemispheric differences in amygdala contributions to response monitoring. AB - The amygdala detects aversive events and coordinates with the rostral anterior cingulate cortex to adapt behavior. We assessed error-related activation in these regions and its relation to task performance using functional MRI and a saccadic paradigm. Both amygdalae showed increased activation during error versus correct antisaccade trials that was correlated with error-related activation in the corresponding rostral anterior cingulate cortex. Together, activation in the right amygdala and right rostral anterior cingulate cortex predicted greater accuracy. In contrast, the left amygdala activation predicted a higher error rate. These findings support a role for the amygdala in response monitoring. Consistent with proposed specializations of the right and left amygdala in aversive conditioning, we hypothesize that right amygdala-rostral anterior cingulate cortex interactions mediate learning to avoid errors, whereas left error-related amygdala activation underpins detrimental negative affect. PMID- 19218866 TI - Dissociated brain potentials for two calculation strategies. AB - Event-related brain potentials were used to investigate the shortcut calculation strategy and nonshortcut calculation strategy in performing addition using mental arithmetic. Results showed that the shortcut calculation strategy elicited a larger P220 than the nonshortcut calculation strategy in the 180-280 ms. Dipole source analysis of the difference wave (shortcut calculation minus nonshortcut calculation) indicated that a generator was localized in the posterior cingulate cortex, which reflected the evaluation effect of number in the use of the shortcut strategy. In the 320-500 ms time window, a greater N400 was found in the nonshortcut calculation as compared with the shortcut calculation. Dipole source analysis of the difference wave indicated that a generator was localized in the anterior cingulate cortex. The N400 might reflect the greater working memory load. PMID- 19218868 TI - Intracerebroventricular infusion of cytosine-arabinoside causes prepulse inhibition disruption. AB - Adult neurogenesis in hippocampus is associated with behaviors such as learning. Hippocampus is involved in the regulation of prepulse inhibition (PPI), but the relationship between neurogenesis and PPI is unexplored. We conducted four experiments to determine the role of neural progenitor cell proliferation in PPI. Intracerebroventricular infusion of cytostatic cytosine arabinoside caused PPI disruption but repeated exposure to PPI sessions prevented the PPI disruption. Corticosterone treatment, which decreases hippocampal cell proliferation, caused PPI disruption, whereas antidepressant and exercise, which increased cell proliferation, did not affect PPI. These results suggest that cell proliferation is involved in the first encounter with PPI test while its importance may decrease upon repeated exposures to the tests. PMID- 19218867 TI - Early (M170) activation of face-specific cortex by face-like objects. AB - The tendency to perceive faces in random patterns exhibiting configural properties of faces is an example of pareidolia. Perception of 'real' faces has been associated with a cortical response signal arising at approximately 170 ms after stimulus onset, but what happens when nonface objects are perceived as faces? Using magnetoencephalography, we found that objects incidentally perceived as faces evoked an early (165 ms) activation in the ventral fusiform cortex, at a time and location similar to that evoked by faces, whereas common objects did not evoke such activation. An earlier peak at 130 ms was also seen for images of real faces only. Our findings suggest that face perception evoked by face-like objects is a relatively early process, and not a late reinterpretation cognitive phenomenon. PMID- 19218869 TI - Phobic spider fear is associated with enhanced attentional capture by spider pictures: a rapid serial presentation event-related potential study. AB - The early posterior negativity (EPN) reflects early selective visual processing of emotionally significant information. This study explored the association between fear of spiders and the EPN for spider pictures. Fifty women completed a Spider Phobia Questionnaire and watched the random rapid serial presentation of 600 neutral, 600 negatively valenced emotional, and 600 spider pictures (three pictures per second). The EPN was scored as the mean activity in the 225-300-ms time window at lateral occipital electrodes. Participants with higher scores on the phobia questionnaire showed larger (i.e. more negative) EPN amplitudes in response to spider pictures. The results suggest that the attentional capture of spider-related stimuli is an automatic response, which is modulated by the extent of spider fear. PMID- 19218870 TI - Epidermal growth factor receptor is involved in clusterin-induced astrocyte proliferation. AB - We previously reported that clusterin enhances astrocyte proliferation and extracellular signal-regulated kinase (ERK) activity. It, however, remains largely unknown how clusterin promotes cell growth. Here, we investigate the signaling pathway and related molecules underlying astrocyte proliferation by clusterin. Exogenous clusterin stimulates Ras-dependent Raf-1/mitogen-activated protein kinase kinase (MEK)/ERK activation. Clusterin-induced astrocyte proliferation and ERK1/2 phosphorylation were abrogated by either AG1478 (an inhibitor of epidermal growth factor receptor, EGFR) or EGFR small interfering RNA. Furthermore, clusterin treatment provoked tyrosine phosphorylation of EGFR (pY(1173)), which was also blocked by AG1478. These results suggest that clusterin requires EGFR activation to deliver its mitogenic signal through the Ras/Raf-1/MEK/ERK signaling cascade in astrocytes. PMID- 19218871 TI - Suppression of multisensory integration by modality-specific attention in aging. AB - Previous research shows that modality-specific selective attention attenuates multisensory integration in healthy young adults. In addition, older adults evidence enhanced multisensory integration compared with younger adults. We hypothesized that these increases were because of changes in top-down suppression, and therefore older adults would show multisensory integration while selectively attending. Performance of older and younger adults was compared on a cued discrimination task. Older adults had greater multisensory integration than younger adults in all conditions, yet were still able to reduce integration using selective attention. This suggests that attentional processes are intact in older adults, but are unable to compensate for an overall increase in the amount of sensory processing during divided attention. PMID- 19218872 TI - Diffusion tensor MRI of the corpus callosum and cognitive function in adults born preterm. AB - Very preterm birth (before 33 weeks gestation) is associated with the white matter damage, and a common sequel is reduced size and altered shape of the corpus callosum. We used diffusion tensor MRI to assess the corpus callosum in 63 very preterm and 45 term-born young adults. Indices of white matter microstructure [fractional anisotropy (FA) and mean diffusivity (MD)] were obtained for the genu, body and splenium. Very preterm females had higher MD in the genu than term-born females, indicating altered white matter microstructure. This was associated with lower performance IQ. The groups demonstrated different patterns of correlations between verbal learning and tract-specific FA and MD, consistent with the reorganization of white matter structure in adults born very preterm. PMID- 19218873 TI - The broken trust and cooperation in borderline personality disorder. AB - Trust and cooperation are essential features of human interpersonal transactions. Recent evidence suggests that these processes are related to brain areas implicated in social decision-making. These novel data provide a unique opportunity to characterize psychopathological conditions in which trust and cooperation are potentially impaired. Using economic games, independent investigations revealed that trust and cooperation are disrupted in patients with borderline personality disorder who have severe difficulties in their personal relationships and exhibit abnormal emotion regulation. Data from functional neuroimaging indicated that the abnormal activation of the anterior insula might be a key factor during these processes, together with the cingulate cortex and the amygdala. PMID- 19218874 TI - Cadherin-20 expression by motor neurons is regulated by Sonic hedgehog during spinal cord development. AB - During chicken spinal cord development, cadherin-20 is expressed by motor neurons of the lateral motor column and other cells in the basal and alar plates. To investigate the regulation of cadherin-20 expression, Sonic hedgehog (Shh) signaling was altered in chicken spinal cord and hindbrain by in-vivo electroporation. Our results show that, at an early embryonic stage 12, Shh induces cadherin-20 expression by motor neurons. Later, at stage 24 when the sorting of motor neuron pools begins, cadherin-20 expression is induced in the neural progenitors of the ventricular zone but not in motor neurons. Blockage of Shh signaling inhibits cadherin-20 expression in the motor column. Therefore, cadherin-20 expression in motor neurons is regulated by Shh in a time-dependent manner. PMID- 19218875 TI - Depressed adolescents demonstrate greater subgenual anterior cingulate activity. AB - Neuroimaging studies implicate the subgenual anterior cingulate cortex (sgACC) as a critical brain region in adult depression. However, unlike adult depression, little is known about the underlying neural substrates of adolescent depression, and there are no published data examining differences in sgACC activation between depressed and healthy adolescents. This study used functional magnetic resonance imaging to examine sgACC activity in 26 depressed and normal 13-17-year olds during the performance of a stop-signal task. Significantly greater sgACC activation was found in the depressed adolescents relative to controls. These results establish for the first time abnormal functioning of the sgACC in depressed adolescents and have important implications for understanding the underlying neural correlates and potential treatments of adolescent depression. PMID- 19218876 TI - Decreased gyrification in major depressive disorder. AB - Structural and functional abnormalities have been extensively reported in major depressive disorder, but possible changes in cortical folding have not yet been explored in this disorder. This study investigated this issue in major depressive disorder using the local gyrification index. High-resolution magnetic resonance imaging was performed in 18 patients with first-episode major depressive disorder and 18 age-matched and sex-matched healthy individuals. The local gyrification index was applied to detect brain areas with abnormal cortical folding in major depressive disorder. Compared with healthy participants, patients with major depressive disorder showed significantly decreased local gyrification index in the bilateral mid-posterior cingulate, insula, and orbital frontal cortices, the left anterior cingulate cortex, and the right temporal operculum. PMID- 19218877 TI - Upregulation of the P2X7 receptor after cryogenic injury to rat brain. AB - The P2X7 receptor is a member of the P2X family of adenosine triphosphate-gated ion channels. It is expressed in the central nervous system and is associated with several pathological conditions. Here, we examined the expression of P2X7 with immunohistochemistry after cryogenic injury to the rat cortex. One day after cortical cryogenic injury, P2X7 immunoreactive cells were increased in regions surrounding the cold-injured site. These P2X7 positive cells were also immunoreactive for OX42, a microglial marker, and possessed the short, thick processes that are characteristic of activated microglia. These results suggest that the increased local expression of P2X7 may be associated with microglial activation, contributing to detrimental and/or protective functions around the cold-injured site. PMID- 19218878 TI - Erythropoietin-receptor gene regulation in neuronal cells. AB - Because erythropoietin (Epo) is intensively studied as neuroprotective agent, Epo receptor (EpoR) regulation in neurons is of particular interest. Herein, we investigated molecular mechanisms of EpoR regulation in neuronal cells including the role of GATA transcription factors. First, developmental downregulation of EpoR expression in murine brain was observed. A differential expression pattern of the Gata factors was found in these specimens as well as in murine adult neural stem cells (NSC) and primary rat neurons, astrocytes, and microglia. Human SH-SY5Y cells served as a model to analyze EpoR regulation. In vitro binding of GATA-2, -3, and -4 to the 5'-flanking region was demonstrated. In reporter gene assays, the activity of a region containing two GATA binding sites was significantly induced when these GATA factors were overexpressed. However, GATA factors alone did not affect endogenous EpoR expression. Importantly, EpoR transcripts have doubled under hypoxia. Furthermore, we analyzed the methylation pattern close to the GATA motifs. Indeed, demethylation with 5-Aza-2' deoxycytidine (Aza) resulted in upregulation of EpoR mRNA. Additionally, several CpGs were mostly nonmethylated in SH-SY5Y cells, but methylated in specific regions of the human adult brain. Thus, methylation may be involved in developmental EpoR downregulation. PMID- 19218879 TI - The T(reg)/Th17 cell balance: a new paradigm for autoimmunity. AB - Regulatory T cells and T helper 17 cells are two recently described lymphocyte subsets with opposing actions. In this review, we discuss the mechanisms that promote development of these cells from common precursors and the specific factors that impact their cell numbers and function. Altered regulation of this key developmental checkpoint may contribute to the pathophysiology of autoimmune diseases by tipping the balance toward inflammation. We also present recent findings that suggest how the equilibrium between regulatory T cells and proinflammatory T helper subsets might be pharmacologically restored for therapeutic benefit. PMID- 19218880 TI - Intermittent hypoxia in supine versus side position in term neonates. AB - Supine sleeping, recommended to prevent sudden infant death, should be started in the delivery hospital, but meets concern that neonates may exhibit more intermittent hypoxia in this position. We aimed to demonstrate that intermittent hypoxia is equally rare in supine and side position. We enrolled 609 term neonates in a crossover study. Pulse oximeter saturation (Spo2) was recorded throughout; feeding periods were excluded. The rate of desaturation events to <80%/h was the primary study variable. Four hundred seventy-six recordings fulfilled study criteria. Mean duration of recording was 4.8 h in side and 4.9 h in supine position (SD 0.8 for both). There was a mean of 0.11 desaturation events per hour in supine and 0.05 per hour in side position. Of the 118 infants with desaturation events, more had such events only in supine position (64 vs. 31 infants). For supine compared with side position, the adjusted odds ratio for at least one desaturation event was 2.0 (95% confidence interval 1.3-3.1). The lowest Spo2 value reached during these events was 56% in supine and 53% in side position. These neonates had more desaturation events while sleeping supine. We speculate, however, that this positional effect likely is too small to affect outcome. PMID- 19218881 TI - Association of preterm birth with brain malformations. AB - This study investigates the rate of preterm birth in babies with congenital brain defects. Autopsy case reports of congenital brain anomalies were obtained from the literature. The control cases were from a large registry, a published report from the Metropolitan Atlanta Congenital Defects Program. From 428 publications, 1168 cases were abstracted that had autopsy studies of congenital brain defects and information on the gestational age (GA) at birth. The control data from Atlanta included 7738 infants with significant birth defects of any kind and 264,392 infants without birth defects. In the autopsy cases with brain defects, the mean GA was 36.6 wk, whereas the Atlanta data showed a mean GA of 39.3 wk for infants with no defects and a significantly shorter gestation of 38.1 wk (p < 0.0001) for infants with defects. In the Atlanta data, the rate of preterm birth was 9.3% for those with no defects compared with 21.5% (p < 0.0001) for those with defects. In the autopsy cases with brain defects, the rate of preterm birth was even greater (33.1%, p < 0.0001). In conclusion, these data show an association of brain defects with preterm births. PMID- 19218882 TI - Placental 11beta-HSD2 gene expression at birth is inversely correlated with growth velocity in the first year of life after intrauterine growth restriction. AB - Intrauterine growth restriction (IUGR) is associated with an increased risk for short stature and diseases in adulthood thought to be inflicted by fetal programming. We hypothesized that placental endocrine systems involved in perinatal growth might also play a role in postnatal growth after IUGR. In a prospective controlled multicenter study, placental gene expression of IGF binding protein-1 (IGFBP-1), leptin and 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2) were measured in 14 IUGR infants and 15 children born appropriate for gestational age (AGA) proven by serial ultrasound examinations. Postnatally, IUGR infants experienced a significantly higher growth velocity than AGA neonates (at 1 y: p = 0.001). Gene expression of 11beta-HSD2 at birth correlated positively with birth length (r = 0.55, p = 0.04) and inversely with growth velocity in the first year of life (r = -0.69, p = 0.01) in the IUGR, but not in the AGA group. There was no correlation between gene expression of placental IGFBP-1, leptin and birth weight, length and growth velocity during the first year of life. AGA infants showed significantly higher concentrations of cortisone in venous cord blood after birth (p = 0.02) as a surrogate of a higher 11beta HSD2 activity in the fetoplacental unit. In conclusion, placental 11beta-HSD2 gene expression might predict postnatal growth in IUGR. PMID- 19218883 TI - Caudal regression and tracheoesophageal malformation induced by adriamycin: a novel chick model of VATER association. AB - VATER association represents a cluster of Vertebral, Anal, Tracheo-Esophageal, Radial and Renal malformations, and caudal regression syndrome is an entity consisting of a spectrum of congenital anomalies of lower spine and hips associated with genitourinary and lower limb defects. The concurrence of various malformations may be explained by a common defect in blastogenesis, but direct evidence is yet to be accumulated. Here, by the use of autofluorescence and the teratogenic effect of adriamycin, we demonstrated that adriamycin administered to eggs of White Leghorns distributes to the caudal portion of the embryo and foregut epithelium and induces caudal regression and tracheal and pulmonary agenesis. The induction of caudal regression syndrome-like anomaly was developmental stage and dose dependent. Embryos with caudal regression demonstrated tracheoesophageal anomalies, one of the defects included in VATER association. The stages at which anomalies were produced corresponded to that of human embryos between days 22 and 26 (Carnegie stages 10-11). In view of the antitumor activity of adriamycin by intercalating to double-stranded DNA of undifferentiated cells undergoing rapid cell division, it is possible that adriamycin had preferentially attacked cells in the caudal end where pronounced proliferation takes place during this narrow period of greatest susceptibility. PMID- 19218884 TI - Patient-ventilator interaction during neurally adjusted ventilatory assist in low birth weight infants. AB - Neurally adjusted ventilatory assist (NAVA), a mode of mechanical ventilation controlled by diaphragmatic electrical activity (EAdi), may improve patient ventilator interaction. We examined patient-ventilator interaction by comparing EAdi to ventilator pressure during conventional ventilation (CV) and NAVA delivered invasively and non-invasively. Seven intubated infants [birth weight 936 g (range, 676-1266 g); gestational age 26 wk (range, 25-29)] were studied before and after extubation, initially during CV and then NAVA. NAVA-intubated and NAVA-extubated demonstrated similar delays between onset of EAdi and onset of ventilator pressure of 74 +/- 17 and 72 +/- 23 ms (p = 0.698), respectively. During CV, the mean trigger delays were not different from NAVA, however 13 +/- 8.5% of ventilator breaths were triggered on average 59 +/- 27 ms before onset of EAdi. There was no difference in off-cycling delays between NAVA-intubated and extubated (32 +/- 34 versus 28 +/- 11 ms). CV cycled-off before NAVA (120 +/- 66 ms prior, p < 0.001). During NAVA, EAdi and ventilator pressure were correlated [mean determination coefficient (NAVA-intubated 0.8 +/- 0.06 and NAVA-extubated 0.73 +/- 0.22)]. Pressure delivery during conventional ventilation was not correlated to EAdi. Neural expiratory time was longer (p = 0.044), and respiratory rate was lower (p = 0.004) during NAVA. We conclude that in low birth weight infants, NAVA can improve patient-ventilator interaction, even in the presence of large leaks. PMID- 19218885 TI - Epidemiology of pervasive developmental disorders. AB - This article reviews the results of 43 studies published since 1966 that provided estimates for the prevalence of pervasive developmental disorders (PDDs), including autistic disorder, Asperger disorder, PDD not otherwise specified, and childhood disintegrative disorder. The prevalence of autistic disorder has increased in recent surveys and current estimates of prevalence are around 20/10,000, whereas the prevalence for PDD not otherwise specified is around 30/10,000 in recent surveys. Prevalence of Asperger disorder is much lower than that for autistic disorder and childhood disintegrative disorder is a very rare disorder with a prevalence of about 2/100,000. Combined all together, recent studies that have examined the whole spectrum of PDDs have consistently provided estimates in the 60-70/10,000 range, making PDD one of the most frequent childhood neurodevelopmental disorders. The meaning of the increase in prevalence in recent decades is reviewed. There is evidence that the broadening of the concept, the expansion of diagnostic criteria, the development of services, and improved awareness of the condition have played a major role in explaining this increase, although it cannot be ruled out that other factors might have also contributed to that trend. PMID- 19218887 TI - Initial validation of a novel protein biomarker panel for active pediatric lupus nephritis. AB - Lupus nephritis (LN) is among the main determinants of poor prognosis in systemic lupus erythematosus (SLE). The objective of this study was to 1) isolate and identify proteins contained in the LN urinary protein signature (PS) of children with SLE; 2) assess the usefulness of the PS proteins for detecting activity of LN over time. Using surface-enhanced or matrix-assisted laser desorption/ionization time of flight mass spectrometry, the proteins contained in the LN urinary PS were identified. They were transferrin (Tf), ceruloplasmin (Cp), alpha1-acid-glycoprotein (AGP), lipocalin-type prostaglandin-D synthetase (L-PGDS), albumin, and albumin-related fragments. Serial plasma and urine samples were analyzed using immunonephelometry or ELISA in 98 children with SLE (78% African American) and 30 controls with juvenile idiopathic arthritis. All urinary PS proteins were significantly higher with active vs. inactive LN or in patients without LN (all p < 0.005), and their combined area under the receiver operating characteristic curve was 0.85. As early as 3 mo before a clinical diagnosis of worsening LN, significant increases of urinary Tf, AGP (both p < 0.0001), and L PGDS (p < 0.01) occurred, indicating that these PS proteins are biomarkers of LN activity and may help anticipate the future course of LN. PMID- 19218888 TI - Biomarker discovery: validation and decision-making in product development. AB - The conference on which this supplement reports was organized to bring together microbicide researchers with researchers in the areas of drug prophylaxis and therapeutics development. The goal of session 1 was to share methods used to validate markers of disease, elucidate the logic used to substantiate the performance of those markers, and identify ways to translate this experience into practical steps for developing microbicides. The experiences discussed ranged from de novo discovery of new therapeutics to qualification of biomarkers across all stages of development, and covered the complexity of biomarker identification, use, and assessment in the clinical areas of cancer and infectious disease. This review is based on the presentations of Drs. Harsukh Parmar (AstraZeneca), Sudhir Srivastava (National Cancer Institute [NCI]), and Juan Leal (Exelixis Inc.), and the subsequent discussion led by Drs. Sam Niedbala (Lehigh University) and Thomas Moench (ReProtect) during a background session held at the conference entitled "Biomarkers for evaluation of vaginal microbicides and contraceptives: Discovery and early validation," organized by CONRAD and the Alliance for Microbicide Development in November 2006. PMID- 19218889 TI - Understanding basic mechanisms and optimizing assays to evaluate the efficacy of vaginal microbicides. AB - The ideal microbicide or microbicide combination must have activity against cell free and cell-associated primary HIV isolates representing multiple clades, must inhibit transmission to relevant cell types within the mucosa, and must retain activity in the presence of cervicovaginal fluid and when virus is introduced in semen or seminal plasma. This summary briefly reviews some of the basic concepts underlying the optimization of assays to evaluate the efficacy of candidate microbicides against HIV/STI. The review is based on the presentations of Drs. Stuart Turville (Population Council), Dan Barouch (Harvard University), Scott McCoombe (Northwestern University), Betsy Herold (Albert Einstein College of Medicine), Sam Niedbala (Lehigh University), and the subsequent discussion led by Drs. Robin Shattock (St. George's Hospital Medical School) and Jim Turpin (National Institutes of Health) during a session entitled "Biomarkers of HIV/STI" held at the conference entitled "Biomarkers for evaluation of vaginal microbicides and contraceptives: discovery and early validation," organized by CONRAD and the Alliance for Microbicide Development in November of 2006 as well as more recent published findings. PMID- 19218890 TI - Biomarkers of cervicovaginal inflammation for the assessment of microbicide safety. AB - The human cervicovaginal mucosa is the primary target of HIV-1 infection during male to female transmission. This tissue contains the the full spectrum of cell types and immune modulators that comprise both the innate and adaptive arms of the immune system. Mounting evidence indicates that mucosal epithelial cells are sentinels of the female reproductive tract, producing innate immune mediators that control the vaginal microflora under normal conditions. Recent studies, however, indicate that certain factors secreted in response to another pathogen or after exposure to a vaginal product may in fact enhance infection by HIV-1. Mucosal inflammation and CD4 cell activation as well as disruption of TLR function and epithelial integrity represent potential causes for such effect. It is therefore important to make sure that vaginal products, including microbicides, do not disrupt the structure or function of the cervicovaginal mucosa. Although a number of biomarkers have been linked to microbicide-induced cervicovaginal inflammation and many of these markers have been measured in preclinical and clinical assays, there are currently no data that demonstrate a correlation between any one marker and susceptibility to HIV-1 infection in humans. To date, the lack of a validated biomarker of cervicovaginal safety represents a gap in the knowledge base that hinders the rational and expeditious selection of microbicide candidates entering clinical trials. Current discovery efforts and preclinical assessment of microbicide safety use an integrated sequential evaluation system that includes cell-based models, explant-based models, and animal-based models. Relevant research in these areas is yielding new assays and biomarkers that, if validated, will be essential to the rational selection of microbicide candidates for efficacy trials. PMID- 19218891 TI - Biomarkers of semen exposure. AB - Biomarkers of semen exposure have historically played their most important role in forensics, i.e., determining whether ejaculation took place in the course of a crime. However, it is becoming increasingly recognized that biomarkers of semen exposure can be useful in the development of new vaginal methods of HIV/sexually transmitted infections (STI) prevention and contraception. This review is based on the presentations of Drs. Michael Coppola (ContraVac, Inc.), Maurizio Macaluso (Centers for Disease Control and Prevention), Andrezej Kulczycki (University of Alabama at Birmingham), Christine Mauck (CONRAD), Robin Maguire (Population Council), and the subsequent discussion led by Drs. Susan Ballagh (CONRAD) and Johan Melendez (Johns Hopkins University) during a session entitled "Biomarkers of semen exposure" held during the conference entitled "Biomarkers for evaluation of vaginal microbicides and contraceptives: Discovery and early validation," organized by CONRAD and the Alliance for Microbicide Development in November of 2006. PMID- 19218892 TI - Biomarkers for evaluating vaginal microbicides and contraceptives: discovery and early validation. AB - A biomarker has been defined as "a characteristic that is objectively measured and evaluated as an indicator of normal biological processes, pathogenic processes, pharmacological responses to a therapeutic intervention." Biomarkers can reduce the costs and time required to get a drug from discovery to market. Topical microbicides are new drugs designed to prevent HIV and other sexually transmitted infections (STIs). Biomarkers that may be important in microbicide development include biomarkers of semen exposure, biomarkers of cervicovaginal inflammation, and biomarkers of HIV and STIs. The development of biomarkers for use in microbicide development is a high priority. This supplement reports on a meeting entitled "Biomarkers for Evaluating Vaginal Microbicides and Contraceptives: Discovery and Early Validation," held in Reston, VA on November 16 to 17, 2006. It was sponsored by CONRAD and the Alliance for Microbicide Development with funding from the Bill and Melinda Gates Foundation and the United States Agency for International Development (USAID). The meeting was convened to look at the ways in which biomarkers could be used to attenuate the challenges alluded to above. Availability of key biomarkers could expedite the development of microbicides, for which there is a pressing need, especially in developing countries where combinations of cultural and socioeconomic pressures on women constrain their ability to protect themselves from STIs. Although the meeting was held 2 years ago, the material reviewed and conclusions drawn are still relevant. PMID- 19218893 TI - Autism spectrum disorders and childhood-onset schizophrenia: clinical and biological contributions to a relation revisited. AB - OBJECTIVE: To highlight emerging evidence for clinical and biological links between autism/pervasive developmental disorder (PDD) and schizophrenia, with particular attention to childhood-onset schizophrenia (COS). METHOD: Clinical, demographic, and brain developmental data from the National Institute of Mental Health (and other) COS studies and selected family, imaging, and genetic data from studies of autism, PDD, and schizophrenia were reviewed. RESULTS: In the two large studies that have examined this systematically, COS is preceded by and comorbid with PDD in 30% to 50% of cases. Epidemiological and family studies find association between the disorders. Both disorders have evidence of accelerated trajectories of anatomic brain development at ages near disorder onset. A growing number of risk genes and/or rare small chromosomal variants (microdeletions or duplications) are shared by schizophrenia and autism. CONCLUSIONS: Biological risk does not closely follow DSM phenotypes, and core neurobiological processes are likely common for subsets of these two heterogeneous clinical groups. Long term prospective follow-up of autistic populations and greater diagnostic distinction between schizophrenia spectrum and autism spectrum disorders in adult relatives are needed. PMID- 19218894 TI - Childhood problem behaviors and death by midlife: the British National Child Development Study. AB - OBJECTIVE: To examine childhood problem behaviors manifested as externalizing behaviors (e.g., aggression, impulsivity) and internalizing behaviors (e.g., anxiousness, avoidant behavior) as predictors of mortality by the age of 46 years and to assess whether these associations are dependent on childhood family background. METHOD: The participants were 5,426 girls and 5,716 boys born in 1958 and participants in the British National Child Development Study (total N = 11,142). Childhood problem behaviors were assessed by teachers at ages 7 and 11 years, and the participants were followed for mortality to the age of 46 years. RESULTS: Both externalizing and internalizing behaviors were associated with mortality in adulthood. By the age of 46 years, the cumulative probabilities of death by increasing externalizing score quartiles were 1.4%, 2.2%, 2.3%, and 3.2%, respectively (odds ratio [OR] for mortality per 1 SD increase in standardized externalizing score 1.27; 95% confidence interval [CI] 1.13-1.44). The corresponding percentages for internalizing score quartiles were 1.8%, 1.9%, 2.3%, and 3.0% (OR 1.20; 95% CI 1.06-1.35). Adjusting for father's social class, family difficulties, family size, and cognitive ability attenuated these associations for externalizing behaviors (OR 1.21; 95% CI 1.06-1.37) and for internalizing behaviors (OR 1.11; 95% CI 0.98-1.26). Childhood environment did not modify the association between problem behaviors and mortality. CONCLUSIONS: Childhood problem behaviors are associated with increased long-term mortality risk beyond childhood and adolescence. PMID- 19218895 TI - ADHD and poor motor performance from a family genetic perspective. AB - BACKGROUND: Attention-deficit/hyperactivity disorder (ADHD) is frequently accompanied by motor problems (MPs). We investigated a possible shared etiology between the two traits in the Dutch sample of the International Multicenter ADHD Genetics study comprising 275 children with ADHD and their affected or unaffected sibling and 146 unrelated control children. METHOD: Exploratory data analysis and bivariate structural equation modeling were used to estimate the familiality of MP rated by parents (Developmental Coordination Disorder Questionnaire [DCD-Q]) or teachers (Groningen Motor Observation Scale [GMO]) and to determine the familial and environmental correlation between MP and ADHD. Furthermore, the nature of the familiality was explored by studying the siblings of ADHD-affected children. RESULTS: The ADHD-affected children had significantly more MP than their unaffected siblings, who in turn had significantly more MP than the control subjects. The familial component of MP measured by DCD-Q and GMO was 47% and 22%, respectively. The familial correlation between motor performance measures and ADHD was -0.38 for DCD-Q and -0.40 for GMO. Our data suggested that co-occurrence of ADHD and MP possibly marks a distinct subtype of ADHD, rather than signaling increased severity of disease. CONCLUSIONS: Attention-deficit/hyperactivity disorder and MP have a common basis that may be due to genetic factors and/or shared environmental factors. Attention-deficit/hyperactivity disorder accompanied by MP may behave like a distinct subtype of ADHD, but more research will be needed to support that hypothesis. PMID- 19218896 TI - Stimulant dosing for children with ADHD: a medical claims analysis. AB - OBJECTIVE: To evaluate stimulant dosing patterns in the community treatment of children with attention-deficit/hyperactivity disorder (ADHD). METHOD: Claims data from U.S. managed care organizations (2000-2004) were analyzed for patients ages 6 to 12 years treated for ADHD with osmotically released oral system (OROS) methylphenidate (MPH; n = 3,815), immediate-release (IR) MPH (n = 1,960), mixed amphetamine salts extended-release (MAS XR; n = 1,847), or IR MAS (n = 1,937), and who filled prescriptions covering at least 72 of the first 90 days of treatment. RESULTS: The mean initial and maximum dosages were 23.8 and 33.4 mg/day for OROS MPH, 14.8 and 21.8 mg/day for IR MPH, 12.7 and 17.4 mg/day for MAS XR, and 11.2 and 16.5 mg/day for IR MAS. Dose titration occurred in 51.8% (MAS XR) to 61.6% (IR MPH) of patients. Lower initial dose and three or more visits for the treatment of ADHD during the first 90 days of treatment were associated with dose titration. Maximum dose was significantly related to higher initial dose and titration for all four stimulants. For children treated with OROS MPH or IR MAS, treatment by a psychiatrist was significantly related to higher maximum dose. CONCLUSIONS: Among children with ADHD who continue stimulants through the first 3 months of treatment, dosing in the community treatment of ADHD tends to be lower than doses used in clinical trials. When titration does occur, it is linked to lower initial dosing, clinical monitoring, higher final stimulant doses, and treatment by a psychiatrist. PMID- 19218897 TI - Hallermann-Streiff Syndrome: a case report from Turkey. AB - Hallermann-Streiff Syndrome (HSS) is a rare disorder characterized primarily by head and face abnormalities, with dental abnormalities also present in 50-80 percent of cases. The first description seems to have been made by Aubry in 1893. HSS was first described completely in 1948 by Hallermann, and then in 1950 by Streiff. We report a 4-year-old girl with HSS, presenting oro-dental characteristics and describe the treatment modalities. An interdisciplinary approach including early preventive-care programmes, detailed oral hygiene instructions, dietary recommendations, counselling of the parents and regular dental visits are the essential procedures. PMID- 19218898 TI - Bilateral bifid mandibular canal: a case report. AB - The objective of this case report is to help clinicians identify bifid mandibular canals on panoramic radiographs and subsequently use the information in the modification of dental treatment planning. A 45-year old man was referred to the service of Oral Diagnosis and Radiology. Routine panoramic radiography, suggested the presence of bilateral bifid mandibular canals (BMC). Mandibular computed tomography revealed a clear view of bilateral mandibular canals. BMC can be detected on a panoramic radiograph. PMID- 19218899 TI - Treatment of comminuted mandibular fractures: a critical review. AB - The treatment of comminuted fractures of the mandible is challenging due both to the severity of the injuries generally associated with this type of fracture, and the lack of consensus as to the most appropriate treatment method.There are two distinct approaches for treating comminuted fractures of the mandible: closed reduction with maxillomandibular fixation (MMF) - the oldest and classical treatment - and open operation and internal fixation. The morbidity rate of closed reduction is lower but, with the advent of modern anaesthesia and antibiotics, open surgery has become more frequent. Stable internal fixation (SIF) is acheived using plates, miniplates and/or screws. The advantage of this approach is that there is a more precise reduction of the fragments, with the possibility of early function by eliminating or reducing the time of MMF. This paper reviews the main advantages, disadvantages and differences between the two techniques. PMID- 19218900 TI - Laser resection of liposarcoma of the hypopharynx. AB - Liposarcomas represent between 15 and 18% of all sarcomas with the most common site being the extremities and retroperitoneum. Liposarcomas of the head and neck are rare, with an estimated incidence representing 3 to 5.6% of all liposarcomas. Liposarcomas most commonly present in the soft tissues of the neck. Primary liposarcoma of the hypopharynx (piriform sinus) is extremely rare. The symptoms presented are principally dysphagia, dyspnea, dysphonia, airway obstruction and sensation of a foreign body. Treatment of choice is surgery, and the literature describes the performance of lateral pharyngotomy, simple excision and even total laryngectomy. We present the case of a 23-year-old patient who was diagnosed 7 years prior with liposarcoma of the piriform sinus. The patient underwent surgery using a cervical approach. The tumor recurred 4 years postoperatively and the patient was again surgically intervened using the same approach. He presented to our Institute with 3 months evolution of dysphonia. Nasofibrolaryngoscopy and imaging studies were performed. Surgical treatment was decided upon with CO2 laser using suspension microlaryngoscopy, obtaining excellent results. Some of the advantages of this approach are low morbidity because of the avoidance of performing a tracheostomy, rapid return to oral feeding without necessity of a feeding tube, and reduction in hospitalization days. Disadvantage includes difficulty in evaluating margins. PMID- 19218901 TI - Comparison of the remineralizing effect of a sodium fluoride mouthrinse versus a sodium monofluorophosphate and calcium mouthrinse: an in vitro study. AB - OBJECTIVES: The aim of this in vitro study was to compare the remineralizing effect of three rinses: (A) 0.17% sodium monofluorophosphate and 0.05% calcium glycerophosphate (220 ppm fluoride), (B) 0.05% sodium fluoride (220 ppm fluoride) and (C) control mouthrinse (without fluoride or calcium). METHOD AND MATERIALS: Demineralized areas were created in 90 pieces of bovine enamel by submerging them in an acid solution (pH 4.4) for 48 hours. Part of the surface was painted with nail varnish to preserve the demineralized area, and the specimens were assigned at random to three groups. The specimens were stored in artificial saliva at pH 7 and 37 degrees C for 30 days. Twice daily they were immersed for 60 seconds in the respective mouthrinse. Following the experimental period, the specimens were bisected and examined by scanning electron microscopy. RESULTS: The mean percentage of remineralization produced by the mouthrinses was as follows: (A) 54.08 (95% CI 46.37-61.78), (B) 38.43 (95% CI 30.89-45.98) and (C) 30.18 (95% CI 25.56-34.80). The differences between the three groups were statistically significant. CONCLUSION: The results show that the fluoride and calcium mouthrinse has a significantly greater remineralizing capacity than the fluoride mouthrinse with the same fluoride ion concentration under the in vitro conditions of this study. PMID- 19218902 TI - Oral granulocytic sarcoma: a case report. AB - Granulocytic sarcoma (GS) is a localized infiltrate of immature granulocytes in an extramedullary site. This lesion is most frequently associated with leukemia, but can occur associated with other myeloproliferative disorders. GS can affect virtually any part of the body, but oral lesions are rare, with only 37 cases described. CASE DESCRIPTION: We present a case of GS in a 23-year-old female, with a prior history of acute myeloid leukemia, presenting with a solitary mandibular swelling in the region of the erupting lower left 3rd molar. After biopsy, conventional immunohistochemical stains were positive for CD45 (hematological marker) and myeloid markers, such as myeloperoxidase, and CD68, demonstrating a myeloid lineage with monocytic cell differentiation, suggesting the diagnosis of GS associated to AML-M5. CLINICAL IMPLICATION: Although GS is a rare tumor in the oral cavity, and its diagnosis is usually difficult, the clinician must know of its existence in order to make a differential diagnosis. PMID- 19218903 TI - Synergistic effects of cigarette smoke and saliva. AB - OBJECTIVES: The aim of this study was to evaluate the cytotoxic effects of Cigarette Smoke on the human peripheral blood lymphocytes in the presence of stimulated or non-stimulated saliva in an in vitro model. METHODS AND MATERIALS: Ten healthy volunteers in the age range of 21 to 29 were selected and samples of peripheral blood lymphocytes and saliva (whole and stimulated saliva) collected . Peripheral blood lymphocytes suspensions (PBS) were taken and exposed to 6 different media. Samples were collected from all media at both 20 and 80 minute time points. The survival rates of PBL were then determined at both 20 and 80 minute time points. RESULTS: The cell survival rates following exposure to cigarette smoke (CS) in the presence of PBS supplemented with whole or stimulated saliva were significantly lower at 80 minutes when compared with the 20 minute rates (P<0.05). CONCLUSION: Tobacco effects were significantly increased in the presence of saliva especially stimulated saliva. PMID- 19218904 TI - Multiple hyperodontia: report of a case with 17 supernumerary teeth with non syndromic association. AB - Supernumerary teeth or hyperodontia is an infrequent developmental alteration that appears in any area of the dental arches and can affect any dental organ. Multiple hyperodontia can be associated with several syndromes such as Gardner's, cleidocranial dysplasia, tricho-rhino phalangic syndrome or in patients with cleft lip and palate. However, it can be present in patients without any systemic pathology. The presence of supernumerary teeth is associated with different alterations in neighboring teeth, being the most common: over retained teeth or delayed eruption, ectopic eruption, dental malposition, occlusal problems, diastemas and rotated neighboring teeth, among a series of associated pathologies. The routine use of radiographic images during examination is a valuable tool that helps make the early diagnosis of these types of abnormalities. Keeping in mind that there are different treatment options, the therapeutic modalities increase. A case of a 20 year old female patient with multiple supernumerary teeth with non syndromic association is presented. The teeth were located in the maxilla and the mandible causing a severe malocclusion. PMID- 19218905 TI - Traumatic bone cyst: a retrospective study of 21 cases. AB - OBJECTIVE: To describe the clinical and radiological characteristics, and surgical findings of traumatic bone cysts. STUDY DESIGN: A retrospective observational study was made of 21 traumatic bone cysts. The diagnosis was based on the anamnesis, clinical exploration, and complementary tests. Panoramic and periapical X-rays were obtained in all cases, together with computed tomography as decided by the surgeon. A descriptive statistical analysis was made of the study variables using the SPSS v12.0 for Windows. RESULTS: There was a clear female predominance (14:7). The mean age was 26.5 years (range 8-45 years). The cysts in all cases constituted casual findings during routine radiological exploration. In those cases where computed tomographic images were available, preservation of the vestibular and lingual cortical layers was observed. Five of the 21 patients (23.8%) reported a clear antecedent of traumatism in the affected zone. All the lesions were subjected to surgery, and the cavities were found to be vacant in 90.5% of the cases. In only two patients were vascular contents seen within the cavity. Two of the patients presented postoperative paresthesia of the inferior dental nerve that subsided within two weeks. The 19 patients in whom adequate postoperative follow-up proved possible all showed complete bone healing. CONCLUSIONS: Traumatic bone cysts were a casual finding. During the surgery, most cases showed to be vacant cavity without an ephitelial lining. Careful curettage of the lesion favors progressive bone regeneration, offering a good prognosis and an almost negligible relapse rate. Other treatment options only would be justified in cases of relapse. PMID- 19218906 TI - Langerhans cell histiocytosis: literature review and descriptive analysis of oral manifestations. AB - Langerhans cell histiocytosis (LCH) is a rare disease, of unknown pathogenesis, characterized by intense and abnormal proliferation of bone marrow-derived histiocytes (Langerhans cells). It can present both local and systemic manifestations involving bone, skin and mucosal tissue, and internal organs. Three basic clinical forms develop: Letterer-Siwe disease (subacute or acute disseminated form), Hand-Schuller-Christian disease (disseminated chronic form) and eosinophilic granuloma (localized chronic form). LCH may manifest orally with single or multiple lesions of the alveolar or basal bone, ulcerated mucosal lesions accompanied by adenopathies and/or periodontal lesions, presenting gingival inflammation, bleeding, recession, necrosis, odontalgia, dental hypermobility and premature loss of teeth. The principal differential diagnoses include advanced periodontal disease or a periapical process of dental or periodontal origin. The odontologist plays a vital role in the diagnosis and multidisciplinary treatment of such patients, by performing routine examinations for periodic follow-up of the disease and its possible oral manifestations, bearing in mind that these may be the first or only signs of LCH. PMID- 19218907 TI - Analysis of new diagnostic methods in suspicious lesions of the oral mucosa. AB - OBJECTIVE: The objective of this study was to analyse publications related to examination techniques that might improve the visualisation of suspicious lesions of the oral mucosa (ViziLite system and VELscope system) or that might facilitate the cytological identification of suspicious lesions (OralCDx). METHODS: A literature search was performed, using the PubMed database and the key words "brush biopsy", "OralCDx", "ViziLite" and "Velscope", limiting the search to papers in English or Spanish published from 2002 to 2008. RESULTS: According to the results of studies identified, the ViziLite system has a sensitivity of 100% and specificity ranging from 0-14.2%, the VELscope system has a sensitivity of 98 100% and specificity of 94-100% and the Oral CDx system has a sensitivity of 71.4 100% and specificity of 32-100%. CONCLUSION: Clinical examination and histopathological confirmation with biopsy remain the gold standard for the detection of oral cancer. More randomised controlled studies are needed to confirm the positive cost-benefit relationship and the true usefulness of these "new diagnostic methods" in oral mucosal pathology. PMID- 19218908 TI - Metastatic malignant melanoma. AB - Metastatic malignant melanoma is an incurable malignancy with extremely poor prognosis. Patients bearing this diagnosis face a median survival time of approximately 9 months with a probability of surviving 5 years after initial presentation at less than 5%. This is contrasted by the curative nature of surgical resection of early melanoma detected in the skin. To date, no systemic therapy has consistently and predictably impacted the overall survival of patients with metastatic melanoma. However, in recent years, a resurgence of innovative diagnostic and therapeutic developments have broadened our understanding of the natural history of melanoma and identified rational therapeutic targets/strategies that seem poised to significantly change the clinical outcomes in these patients. Herein we review the state-of-the-art in metastatic melanoma diagnostics and therapeutics with particular emphasis on multi-disciplinary clinical management. PMID- 19218909 TI - Metallothionein-overexpression as a prognostic marker in melanoma. AB - Malignant melanoma is one of the most aggressive human neoplasms and its incidence is still increasing. Prognoses for melanoma patients are currently based on statistical parameters. For estimating the risk for a possible progression and for overall survival, Breslow tumour thickness and the invasion level (Clark level) are the most established markers for melanomas at the time of the primary diagnosis. In thicker melanomas (>1 mm) the additional information about the status of the sentinel lymph-node (SLN) might be helpful. Nevertheless new prognostic parameters are needed, that will allow us to formulate more precise prognoses for the individual cases. The metallo-thionein family is a class of intracellular low-molecular-weight, cysteine-rich proteins with a high affinity for heavy-metal ions. They are involved in many (patho-) physiological processes and presumably play an important role in the carcinogenesis. In the last decades overexpression of immunohistochemically labelled metallothioneins (MTs) on paraffin-embedded tissues turned out as a highly significant prognostic marker in different tumours. This review summarizes the results of those studies, in which MT-overexpression was able to show a very high significance for progression and survival in melanoma patients. In contrast to most other progression markers, MT-overexpression is independent from tumour-thickness, and is highly specific even in thin (low risk) melanoma patients. Nowadays, in high risk melanoma patients sentinel lymph-node (SLN-) biopsy is performed, a surgical technique with predictive value for progression, the benefit of this procedure for the individual overall survival still remains unclear. In a study comparing SLN and MT-overexpression the results corroborate the validity of MT overexpression in primary melanoma as a useful additional prognostic marker, accuracy is comparable although to some degree supplementary to the results of SLN biopsy. PMID- 19218910 TI - Melanoma progression, gene expression and DNA microarrays. AB - In recent years DNA microarray analyses of gene expression changes in melanoma have been employed in an effort to better understand the processes of disease progression. Typically, the samples assessed are taken directly from tissue biopsies of distinct clinical stages. Apart from noting a significant shift in gene expression at the transition from thin to thick primary lesions, surprisingly little else has been learned. Furthermore, experiments performed using cell cultures derived from distinct clinical stages have entirely failed to identify a consistent stage-specific gene expression signature. This review compares the two approaches, discusses what was learned about the molecular nature of melanoma progression, and considers why stage-specific gene expression may be more complicated than originally thought. PMID- 19218911 TI - Dermoscopy of superficial spreading melanoma. AB - Knowledge and insights gained over the past few decades pertaining to the clinical and dermoscopic primary morphology of melanoma has greatly increased the authors' appreciation of the varied faces of this malignancy. This knowledge has improved their ability to detect early melanoma and may in part explain the observed increase in the percentage of thin melanomas being diagnosed today as compared to the past. The authors have previously published in this journal an article on the dermoscopic patterns of melanoma. In this review they will focus on specific dermoscopic structures that are frequently observed in the most common subtype of melanoma, the superficial spreading melanoma. PMID- 19218912 TI - Chemotherapeutics, chemoresistance and the management of melanoma. AB - Melanoma is one of the most aggressive forms of cancer with a continuously growing incidence worldwide. Survival mainly depends on primary tumor thickness, ulceration and sentinel node status at the time of diagnosis. Adjuvant therapies with interferons are able to prolong the recurrence-free survival, but the effects on overall survival are limited. Once, melanoma has metastasized to distant sites, the prognosis is fatal with median survival times between 7 and 9 months. Albeit removal of localized distant metastases is currently the most effective approach in metastatic melanoma in particular cases, chemo- and chemoimmunotherapy has to be regarded as standard treatment in the majority of patients. However, all available cytotoxic drugs and combinations applied so far have only a small impact on overall survival, if any. A fundamental cause of the limited efficacy of chemotherapy in advanced melanoma has to be seen in chemoresistance mechanisms. In melanoma, the intrinsic and mainly anti-apoptotic resistance, due to the physiological role of the UV exposed melanocytes, is prevailing. Further resistance mechanisms discussed in melanoma are DNA repair, multidrug transporter and the existence of cancer stem cells. Promising therapeutic options accrue from the growing insights into signaling pathways of melanoma that cause chemo- and apoptosis-resistance. The development of drugs targeting those mechanisms and their administration in combination with chemotherapy is currently one of the fascinating novel treatment approaches in melanoma. PMID- 19218913 TI - Photodynamic treatment for viral infections of the skin. AB - Photodynamic therapy (ALA/MAL-PDT) is indicated for the treatment of actinic keratoses, for superficial, nodular basal cell carcinoma and for Bowen's disease; there is evidence that PDT can be active also against bacteria, viruses and fungi. The new indications for PDT include many types of viral skin infections human papilloma virus (HPV)-related as verrucae of feet and hands, Condylomata acuminata, periungueal warts, epidermodysplasia verruciformis, but also viral skin lesions non HPV related as molluscum contagiosum and herpes simplex can be successfully treated. The use of PDT in HPV infections is due to its anti inflammatory and antiproliferative skills: in the lesions treated there is a release of cytotoxic radicals which damage keratinocytes infected by HPV, inducing their selective apoptosis and necrosis. The PDT application in this field of lesions is safe and successful; in comparison with the other techniques it has less side-effects and less recurrences, but the most important property is that it is not-invasive: it means a reduced risk of infections and excellent cosmetic results. PMID- 19218914 TI - Aloe vera in dermatology: a brief review. AB - Aloe vera Linne or aloe barbadensis Miller is a succulent from the Aloe family (400 different species), a tropical plant which is easily grown in hot and dry climates and widely distributed in Asia, Africa and other tropical areas. The use of aloe vera is being promoted for a large variety of conditions. The aim of this systematic review was to summarize all dermatology-oriented in vitro and in vivo experiments and clinical trials on aloe vera preparations. Extensive literature search were carried out to identify all in vitro and in vivo studies as well as clinical trials on the subject. Data were extracted from these in a predefined standardized manner. Forty studies were located. The results suggest that oral administration of aloe vera in mice is effective on wound healing, can decrease the number and size of papillomas and reduce the incidence of tumors and leishmania parasitemia by >90% in the liver, spleen, and bone marrow. Topical application of aloe vera is not an effective prevention for radiation-induced injuries and has no sunburn or suntan protection. It can be effective for genital herpes, psoriasis, human papilloma virus, seborrheic dermatitis, aphthous stomatitis, xerosis, lichen planus, frostbite, burn, wound healing and inflammation. It can also be used as a biological vehicle and an anti-microbial and antifungal agent and also as a candidate for photodynamic therapy of some kinds of cancer. Even though there are some promising results with the use of aloe vera for diverse dermatologic conditions, clinical effectiveness of oral and topical aloe vera is not sufficiently and meticulously explored as yet. PMID- 19218915 TI - Erlotinib-induced pustular eruption. AB - The epidermal growth factor receptor (EGFR) inhibitor erlotinib was found to significantly improve overall survival, time to progression, and cancer-related symptoms in locally advanced or metastatic non small cell lung carcinoma (NSCLC). Unfortunately, cutaneous side effects are not rare, the most common one being a follicular acneiform eruption. As it has been observed a positive correlation between rash severity and objective tumor response, it is critical for dermatologists early distinguish it from rashes carrying a different prognosis and needing a different management. PMID- 19218916 TI - Chemotherapy-induced persistent serpentine supra-venous hyperpigmented eruption and persistent supra-venous erythematous eruption: case report. AB - This paper presents the case of an 85-year-old male affected by classic Kaposi's sarcoma (CKS) since 1994. The disease was widespread over much of the skin surface. Since 1995 the patient had undergone several chemotherapy treatments with good, but not lasting results. Due to a worsening in the pathology, in October 2005 the patient was prescribed a further cycle of vinorelbine. Twenty four hours after the first infusion, followed by isotonic sodium chloride solution vein wash, a supra-venous serpentine erythematous lesion appeared directly above infusion site. The lesion extended centripetally from the injection point and involved the whole supra-venous area. Over the following days, a gradual variation in lesion colour, from erythematous to hyperpigmented, was observed. Given this clinical and histological picture, the diagnosis of persistent serpentine supra-venous hyperpigmented eruption (PSSHE) was made. Draft of a rare secondary effect whose pathogenesis still remain to explain. The literature holds reports of another skin manifestation with similar supra-venous characteristics: persistent supra-venous erythematous eruption (PSSE), clinical entity that enters in differential diagnosis with the PSSHE. Many of the drugs used in chemotherapy have been indicated as responsible for these peculiar side effects. To the authors' knowledge, the literature reports only one PHSSE case induced by i.v. vinorelbine infusion. PMID- 19218917 TI - Lichen amyloidosus. PMID- 19218918 TI - The NS3 protein of rice hoja blanca virus complements the RNAi suppressor function of HIV-1 Tat. AB - The question of whether RNA interference (RNAi) acts as an antiviral mechanism in mammalian cells remains controversial. The antiviral interferon (IFN) response cannot easily be distinguished from a possible antiviral RNAi pathway owing to the involvement of double-stranded RNA (dsRNA) as a common inducer molecule. The non-structural protein 3 (NS3) protein of rice hoja blanca virus (RHBV) is an RNA silencing suppressor (RSS) that exclusively binds to small dsRNA molecules. Here, we show that this plant viral RSS lacks IFN antagonistic activity, yet it is able to substitute the RSS function of the Tat protein of human immunodeficiency virus type 1. An NS3 mutant that is deficient in RNA binding and its associated RSS activity is inactive in this complementation assay. This cross-kingdom suppression of RNAi in mammalian cells by a plant viral RSS indicates the significance of the antiviral RNAi response in mammalian cells and the usefulness of well-defined RSS proteins. PMID- 19218919 TI - A model for DNA replication showing how dormant origins safeguard against replication fork failure. AB - Replication origins are 'licensed' for a single initiation event before entry into S phase; however, many licensed replication origins are not used, but instead remain dormant. The use of these dormant origins helps cells to survive replication stresses that block replication fork movement. Here, we present a computer model of the replication of a typical metazoan origin cluster in which origins are assigned a certain initiation probability per unit time and are then activated stochastically during S phase. The output of this model is in good agreement with experimental data and shows how inefficient dormant origins can be activated when replication forks are inhibited. The model also shows how dormant origins can allow replication to complete even if some forks stall irreversibly. This provides a simple explanation for how replication origin firing is regulated, which simultaneously provides protection against replicative stress while minimizing the cost of using large numbers of replication forks. PMID- 19218920 TI - SASP reflects senescence. PMID- 19218921 TI - Genomic imprinting in Singapore. Workshop on Genomic Imprinting. PMID- 19218922 TI - Are oral health and dentistry compatible? PMID- 19218923 TI - Somewhat amused. PMID- 19218924 TI - Over-prescribing. PMID- 19218925 TI - Expanding, improving. PMID- 19218926 TI - A reliable tool. PMID- 19218927 TI - Can you help? PMID- 19218928 TI - Wikipedia use. PMID- 19218929 TI - Somewhat distressing. PMID- 19218930 TI - Best antibiotic. PMID- 19218931 TI - Undesirable rhythms. PMID- 19218932 TI - HIV positive dentists permitted to practice in Europe. PMID- 19218936 TI - Dental innovators lead the way. PMID- 19218944 TI - Challenges to future dental education. AB - The future of dental education in the UK is currently in a very dynamic phase; the GDC is changing its role and Europe is taking the lead. In addition there have been significant changes in the workforce and the role of the dentist. These changes will affect the future of the dental profession. PMID- 19218945 TI - Case report of self-injurious behaviour (SIB) presenting as gingivitis artefacta major. AB - The case report described here discusses gingivitis artefacta major, an oral presentation of self-injurious behaviour, in an adolescent. On presentation, the patient knew well the ramifications of her gum scratching behaviour, however, was unable to stop. At further presentations new lesions had appeared with further bone loss. The cause of her behaviour seemed to be of psychological origin and therefore no interventive dental treatment was possible until this issue was resolved. A more preventive approach was adopted in the meantime. Referral to appropriate services from the dental profession also proved to be challenging. In conclusion, gingivitis artefacta, although rarely seen to this extent, is extremely challenging to diagnose and treat fully in a dental setting. PMID- 19218946 TI - A clinical guide to endodontics--update part 2. AB - This second update paper aims to provide a review of changes and developments that have occurred within the field of pulp therapy for primary teeth since the publication of the BDJ's series and textbook A clinical guide to endodontics. The biological basis of pulp therapy is an area where much recent research has taken place, particularly linked to the healing capacity of pulp tissue. This has lead to the adoption of treatment techniques aimed at maintaining pulp vitality. Such techniques are introduced here as a more contemporary, biological approach to pulp management in the primary dentition. PMID- 19218947 TI - Immediate implant supported bridgework simultaneous with jaw reconstruction for a patient with mandibular osteosarcoma. AB - A patient with mandibular osteosarcoma underwent full immediate dental rehabilitation including insertion of dental implants and immediately loaded implant bridgework in the same operation as surgical resection of the tumour and scapular composite free flap reconstruction. Planning and pre-production of the titanium reconstruction plate, drill guides and bridgework using a 3D stereolithographic model of the patient's jaw is described. The advantage of this immediate full rehabilitation of an oncology patient is compared with the potential disadvantages. PMID- 19218954 TI - Podcasts--an adjunct to the teaching of dentistry. AB - The aim of this study was to evaluate how clinical students reacted to the use of Podcasts for undergraduate teaching. The most popular way of listening to them was on a computer. Students scored highly the subject matter of the Podcasts and used them for learning around the subject, especially during examination time. Podcasts are a valuable vehicle for delivering learning materials to students: offering flexibility but in tune with students' love of downloading materials to use when they wish. PMID- 19218955 TI - The impact of teamworking on the knowledge and attitudes of final year dental students. AB - AIMS: To relate final year UK dental undergraduates' experience of teamwork related training to their knowledge of the clinical role of dental hygienist therapists, and their views of the clinical roles of dental care professionals. METHODS: A postal questionnaire was sent to the ten UK dental schools which had agreed to participate. Analysis was based on standard non-parametric statistical tests. RESULTS: Of the 705 final year students in the ten schools, 358 (51%) returned questionnaires. Numbers from each school ranged from 79 to 6 (mean 35.8). Eighty-nine percent of students reported receiving information on roles of dental team members; 61% had experienced joint teaching sessions with DCPs; 53% had shared clinical treatment of patients with student hygienists or hygienist therapists. Thirty-two percent of respondents had experienced all three elements. Views on this experience were generally positive, particularly in relation to shared clinical treatment sessions. Students who had received such training were more knowledgeable about the clinical remit of dental hygienist-therapists. However, training experience was not related to their views on the appropriate clinical activity of DCPs which tended to be dentist-centric even for many procedures within the legal remit of hygienists and hygienist-therapists. CONCLUSIONS: The study suggests that acceptance of non-dentists providing patient care lags behind the comparable situation within the primary care medical team. If we are to succeed in the delivery of a modernised dental care system, it is crucial that dental education promotes awareness and acceptance of the professional status and ability of DCP colleagues. PMID- 19218980 TI - The cytotoxic and antifungal activities of two new sesquiterpenes, malfilanol A and B, derived from Malbranchea filamentosa. PMID- 19218981 TI - Microbial degradation of cyclic peptides produced by bacteria. AB - Bacterial strain, B-9, isolated from Lake Tsukui, Japan, and characterized as genus Sphingosinicella sp., possesses hydrolytic enzymes capable of degrading various toxic and non-toxic cyanobacterial cyclic peptides, such as microcystins, nodularin, microviridin, microcyclamide and aeruginopeptin. In this study, the degradation activities of the cell extract of B-9 against bacterial cyclic peptides, bacitracin, colistin, polymyxin, mikamycin, thiopeptin and WAP-8294A2, were investigated and the degradation products were analyzed using HPLC and liquid chromatography/ion trap tandem mass spectrometry (LC/ITMS). As a result of extensive experiments, it was confirmed that B-9 could also degrade these bacterial cyclic peptides by hydrolysis of their peptide or ester bonds, except for WAP-8294A2. These results indicated that the functions of the bacterium with its enzymes were further extended and offered the possibility of degrading other types of compounds. PMID- 19218982 TI - Aldgamycin I, an antibacterial 16-membered macrolide from the abandoned mine bacterium, Streptomyces sp. KMA-001. PMID- 19218983 TI - Synthesis and biological properties of 4''-O-acyl derivatives of 8a-Aza-8a homoerythromycin. AB - A series of 4''-O-acyl derivatives of 8a-aza-8a-homoerythromycins A were synthesized and tested against Gram-positive and Gram-negative bacteria. Derivatives of 8a-aza-8a-homoerythromycin A have potent anti-bacterial activity against not only azithromycin-susceptible strains, but also efflux (M) and inducible macrolide-lincosamide-streptogramin-resistant Gram-positive pathogens. These compounds show moderate to high clearance and low oral bioavailability in preliminary in vivo pharmacokinetic studies in rat. PMID- 19218984 TI - Intravitreal bevacizumab for macular oedema secondary to branch retinal vein occlusion: more data required. PMID- 19218985 TI - Long-term follow-up of children with benign abducens nerve palsy. AB - PURPOSE: Benign abducens nerve palsy is rare in childhood. Diagnosis is made by exclusion, and the severe underlying pathologies have to be ruled out. The aim of our study was to present the largest single-center series of patients with the longest period of follow-up to confirm the benign nature of this entity. PATIENTS AND METHODS: We carried out a retrospective study of 12 consecutive children with benign abducens nerve palsy. All children underwent a careful orthoptic and ophthalmic examination during acute presentation and follow-up. RESULTS: Painless palsies were associated with a preceding infection or immunization in five patients. The left eye was affected in nine children and no bilateral case was found. No sex differences were seen. Recovery was observed within 6 months in all cases, and ipsilateral recurrences occurred in three children. Three children required strabismus surgery. None of the patients developed long-term recurrences or neurological abnormalities during a mean follow-up of more than 9 years. CONCLUSIONS: Our data support earlier findings, such as painless and predominately left-sided occurrence, spontaneous recovery within 6 months, and ipsilateral recurrence. In contrast to much of the literature, we did not find a female preponderance. Exclusion of severe causes and close follow-up is mandatory for these patients. As none of the patients developed long-term recurrences or neurological sequelae, this entity can be regarded as a benign condition without malignant associations or complications. PMID- 19218986 TI - Expression of two endoplasmic reticulum stress markers, GRP78 and GADD153, in rat retinal detachment model and its implication. AB - PURPOSE: To investigate the expression of endoplasmic reticulum (ER) stress related genes, glucose-regulated protein 78 (GRP78) and growth arrest DNA damage inducible gene 153 (GADD153)/CPEBP homologous protein (CHOP), in rat retinal detachment (RD) model. MATERIALS AND METHODS: At various time points after RD, the apoptosis of retinal cells was detected by TdT-mediated fluorescein-16-dUTP nick-end labelling (TUNEL) assay; GRP78 and GADD153 mRNA levels were detected by reverse transcription (RT)-PCR; proteins were detected by western blotting analysis; protein distributions in the retinal cells were observed by immunofluorescence using laser-scanning confocal microscope. RESULTS: After RD, the apoptosis was peaked on 2-4 d and then dropped down. The GRP78 mRNA and GADD153 mRNA levels in RD groups on 0.5, 1, 2, and 4 d were all significantly higher than those in the control group (P<0.05). The expression of GRP78 mRNA peaked on 1-2 d after RD. Expression of GRP78 protein was significantly higher than that in the normal control group on 0.5, 1, 2, 4, 8, 16, and 32 d after RD (P<0.05). The expression of GRP78 protein was observed in all the layers of retina in the RD groups, and peaked on 8, 16, and 32 d. The expression of GADD153 protein, mostly in photoreceptor layers, was significantly higher than that in the control group on 0.5, 1, 2, and 4 d after RD (P<0.05). CONCLUSIONS: ER stress related markers, GRP78 and GADD153, are elevated after RD. The elevation of GADD153 is in parallel with the post-RD apoptosis of retinal cells, suggesting that ER stress-mediated death is likely to be activated after RD and involved in post-RD vision loss. PMID- 19218987 TI - Re: The use of Medpor-coated tear drainage tube in conjunctivodacryocystorhinostomy. PMID- 19218988 TI - Orbital venous-lymphatic malformation. PMID- 19218989 TI - Ocular complications of laser-assisted eyebrow epilation. AB - PURPOSE: To report a series of patients with ocular complications associated with laser-assisted eyebrow hair removal. PATIENTS AND METHODS: Case reports of three patients with eye pain and photophobia following laser epilation of the eyebrow region. The eye examination included visual acuity, slit-lamp examination, tonometry and fundoscopy. The follow-up period was 3 months. RESULTS: Each patient had conjunctival hyperaemia in one or both eyes and anterior chamber pigmentary cells. One patient presented with posterior synechiae, which did not respond to treatment. CONCLUSIONS: Laser epilation of the eyebrows may result in anterior uveitis as well as irreversible damage to the iris. PMID- 19218990 TI - Molecular and immunocytochemical identification of coxsackievirus A-24 variant from the acute haemorrhagic conjunctivitis outbreak in Taiwan in 2007. AB - AIMS: Acute haemorrhagic conjunctivitis (AHC) is a highly contagious conjunctivitis associated with enteroviruses. Coxsackie A-24 variant (CA-24v) and enterovirus-70 (EV-70) are the two major causative agents. During October 2007, an epidemic of AHC occurred in Taiwan, affecting more than 11 000 people. The aim of this study was to determine the aetiological agent associated with the outbreak in patients diagnosed with AHC and treated at the Cathay General Hospital, Taipei (CGHT) and Cathay General Hospital Sijhih (CGHS), Taiwan during October 2007. METHODS: Virus isolates were obtained from six patients (four from CGHS and two from CGH), and a total of seven specimens (one throat and one rectal, and five eye swabs) were collected. The specimens were inoculated onto the MRC-5 cell lines. The viral isolation was confirmed by performing real-time reverse transcription-polymerase chain reaction (RT-PCR) and indirect immunofluorescence assay (IFA). RESULTS: The conjunctival, throat, and rectal swabs collected in this study were all tested positive for a variant of CA-24. All seven viral isolates were characterized as a variant of CA-24 and confirmed by IFA and real-time RT-PCR. CONCLUSIONS: The findings suggest that the outbreak of AHC that occurred during October 2007 in the northern area of Taiwan was caused by a variant of coxsackie A-24. Further phylogenic analysis is underway to further classify this CA-24v strain. PMID- 19218991 TI - Intravitreal triamcinolone acetonide vs bevacizumab for treatment of macular oedema due to central retinal vein occlusion. AB - PURPOSE: To compare the efficacy of intravitreal triamcinolone acetonide vs intravitreal bevacizumab in eyes with macular oedema caused by central retinal vein occlusion (CRVO). DESIGN: Retrospective consecutive case series. METHODS: Retrospective review of the medical records of 35 consecutive patients (35 eyes) with macular oedema associated with CRVO. Twenty-two patients were treated with intravitreal injection of 4 mg/0.1 ml triamcinolone acetonide. The other 13 patients accepted intravitreal bevacizumab 1.25 mg in 0.05 ml. Initial visual acuity, intraocular pressure (IOP), and macular thickness were recorded. Final visual acuity, IOP, macular thickness, and adverse events were recorded during the treatment period. RESULTS: The mean follow-up was 282.73+/-70.62 days in the group administered with triamcinolone acetonide and 253.92+/-36.10 days in the study group who accepted bevacizumab, respectively. Visual acuity measurements improved significantly and showed significant macular oedema resolution in optical coherence tomography examination in both the two groups. However, the therapeutic effects had no significant difference between these two groups with regard to visual results (F=1.723, P=0.240) and macular thickness decrease (F=1.814, P=0.832). Thirteen eyes developed recurrent macular oedema and received repeat injections of triamcinolone acetonide or bevacizumab. CONCLUSION: Intravitreal injection of triamcinolone acetonide or bevacizumab can both lead to a significant improvement in visual acuity and a resolution of macular oedema in patients with CRVO. However, the significant effect was not permanent. Besides, the efficacy of intravitreal triamcinolone acetonide showed no significant differences compared with intravitreal bevacizumab but seemed to cause more adverse events than bevacizumab. PMID- 19218992 TI - Optic nerve head changes in early glaucoma: a comparison between stereophotography and Heidelberg retina tomography. AB - OBJECTIVE: To evaluate and compare the diagnostic accuracy of the Heidelberg retina tomograph (HRT) version 3 with that of glaucoma specialists using stereophotography in discriminating between normal eyes and patients with early glaucomatous visual field loss. METHODS: A total of 105 eyes of 105 individuals were prospectively and consecutively recruited. The sample comprised 51 normal and 54 early glaucomatous eyes, as defined by intraocular pressure and standard automated perimetry results, regardless of optic disc appearance. Receiver operating characteristic (ROC) curves were plotted for the HRT3 parameters and a linear discriminant function (LDF) developed in our hospital. Best sensitivity specificity pairs were compared between the HRT3 parameters, with the highest areas under the ROC curve (AUCs) and evaluation of optic disc stereophotographs. Agreement between methods for measuring vertical cup-to-disc ratio was evaluated with the Bland-Altman plot. RESULTS: The average visual field mean deviation was 2.90 dB. The HRT3 parameters with the largest AUCs were our LDF (0.900), rim volume (0.883), and vertical cup/disk ratio (0.880), with no significant differences between these parameters. Sensitivity-specificity pairs were 79.6 100% (clinical evaluation), 83.3-86.3% (our LDF), 64.8-96.1% (final glaucoma probability score), and 68.5-90.2% (global Moorfields regression analysis). CONCLUSIONS: The diagnostic accuracy for differentiating normal eyes from those with early visual field defects was similar between clinical evaluation of the optic disc and evaluation with the HRT3. The use of our LDF increased the sensitivity-specificity balance with respect to the HRT-provided parameters. The diagnostic accuracy of the HRT classifications was comparable to that of an experienced glaucoma specialist. PMID- 19218993 TI - Phenotypic progression in X-linked retinitis pigmentosa secondary to a novel mutation in the RPGR gene. AB - PURPOSE: To report phenotypic progression for a novel mutation in the RPGRgene causing X-linked retinitis pigmentosa (RP), and describe the phenotype in affected males and females. METHODS: Bidirectional fluorescent sequencing analysis was used to screen for mutations in RPGR. Five affected males and eight affected females from two English families underwent refraction, ETDRS visual acuity, OCT imaging, and Goldmann visual field testing. RESULTS: DNA analysis identified a novel c.350G>A sequence change in exon 5 of RPGR. The change segregated with disease in both families. For affected males there was a significant correlation between age and visual acuity (r=-0.91, P=0.034), and a non-significant correlation between age and visual field area (r=-0.56, P=0.4). For affected females, there was a significant correlation between age and visual acuity (r=-0.8, P=0.018), and between age and visual field area (r=-0.94, P=0.005). All affected females were highly myopic. No correlation between retinal thickness, and either age or sex was noted. CONCLUSION: This novel mutation in RPGRcauses X-Linked RP with complete penetrance in males and females. Affected females are highly myopic but retain better visual function than affected males. The phenotypic data can be used to provide a mutation-specific visual prognosis, and may also help recognition of the genotype. PMID- 19218994 TI - Incidence and management of acute endophthalmitis after intravitreal bevacizumab (Avastin) injection. AB - INTRODUCTION: The aim of this study was to report the incidence and management of acute endophthalmitis after intravitreal injection of Avastin (bevacizumab), and visual acuity outcomes of three eyes of three patients who developed acute endophthalmitis following intravitreal injection of Avastin. METHODS: This clinical retrospective, non-comparative study included 3022 intravitreal injections of 1.25 mg bevacizumab consecutively performed for 1822 eyes with exudative age-related macular degeneration and other retinal diseases. Of 3022 injections, 1200 were reinjections. After clinical appearance of post-injection endophthalmitis, immediate intervention was performed, including injection of intravitreal antibiotics and early pars plana vitrectomy. RESULTS: Three eyes of three patients with acute postoperative endophthalmitis were identified in the first week following intravitreal injections of 1.25 mg bevacizumab. Among of these patients, two cases were culture-positive and one case was culture negative. Compared with presenting visual acuities, all of three patients improved at the end of follow-up time. The overall incidence rate of post injection culture-proven endophthalmitis was 0.066%. DISCUSSION: Acute culture proven endophthalmitis is still a potential complication of intravitreal bevacizumab injection (approximately 0.066%) despite using maximal sterile techniques. Acute post-injection endophthalmitis following intravitreal bevacizumab occurs rapidly and can result in severe loss of vision. Prompt recognition and treatment are key in maximizing outcomes in patients who developed endophthalmitis after intravitreal injection of bevacizumab. PMID- 19218995 TI - Accuracy of corneal flap thickness achieved by two different age MK-2000 microkeratomes. AB - AIMS: To investigate the accuracy of corneal flap thickness (FT) using two different age MK-2000 microkeratomes. METHODS: The prospective cohort study enroled 260 patients with refractive error. Flaps were created using two microkeratomes A and B (new and aged, respectively) with 130-mum heads in two patient groups and two times the same blade in both treated eyes of each patient. The variations in FTs were compared between two groups and between both operated eyes of each patient. The correlations were analysed between FT and CCT or keratometric power. RESULTS: In the A and B groups, the average FTs were 123.3+/ 18.7 and 147.5+/-19.1 mum respectively. Difference in measurements between the actual FTs of first eye operations in the A group and intended 130 mum of FTs was not significant (P=0.462), but those of second operated eyes in the A group and both treated eyes in the B group were significant (P<0.001). Second cut achieved a thinner flap and increased the variability in FT, and an aged microkeratome achieved a thicker flap than a new microkeratome and than that claimed by the manufacturer. Positive correlations were observed between preoperative CCT and FT (P<0.05). CONCLUSIONS: The first eye operation by a new MK-2000 microkeratome achieves the accuracy of the intended FT. FTs varied between first and second cuts of each patient and between two different age MK-2000 microkeratomes. LASIK surgeons should compare FT when using an aged MK-2000 microkeratome, and frequent and periodic comparison of FT achieved by all microkeratomes may be also recommended. PMID- 19218998 TI - The message for World Kidney Day 2009: hypertension and kidney disease: a marriage that should be prevented. PMID- 19219000 TI - Flying podocytes. AB - Recent insights have defined the central role of podocytes both in rare genetic diseases and as a general determinant of the progression of human glomerular diseases. In a recent issue of Nature, Weavers et al. described a podocyte-like cell type in Drosophila, the nephrocyte, that closely resembles mammalian podocytes, including the nephrin-based slit diaphragm. This novel podocyte system might open new avenues toward the understanding of podocyte biology and pathophysiology. PMID- 19219001 TI - The ubiquitin-proteasome pathway and IgA nephropathy: a novel link? AB - Proteasomal degradation of intracellular proteins facilitates the recognition of foreign proteins through interactions with major histocompatibility complex molecules presented to T cells. Interferon-gamma enhances the efficiency of this antigen presentation process by inducing a switch of proteasome to immunoproteasome catalytic subunits. The finding that immunoproteasomes are upregulated in IgA nephropathy may prompt further exploration of the role and mechanism of proteasome activation and intensify the quest for infectious agents that may induce IgA nephritis. PMID- 19219002 TI - Who and where is the renal baroreceptor?: the connexin hypothesis. AB - Gap junctions are emerging as a fundamental mechanism for the control of renin synthesis and release. Connexin40 is prominent in juxtaglomerular cells. When missing, it results in hyperreninemia and hypertension. Schweda et al. offer exciting data demonstrating that connexin45, a connexin with different biophysical properties, can replace connexin40 functions related to the control of renin. PMID- 19219003 TI - Engineering parathyroid cells to treat secondary hyperparathyroidism. AB - Kanai et al. used antisense technology to reduce excessive PTH production. The authors have overcome technical difficulties to demonstrate that, by strategies of RNA interference, a steady reduction of PTH secretion can be induced in cultured parathyroid-cell spheroids and in athymic nude mice with hyperplastic parathyroid cells transplanted from patients with secondary hyperparathyroidism. PMID- 19219005 TI - The case mid R: A kidney transplant presenting with acute renal failure and mass. PMID- 19219006 TI - Nephrology image: quadriceps rupture. PMID- 19219008 TI - Screening we can believe in. AB - Providers and users of chemical libraries must adopt quality and reporting standards to advance the impact of small-molecule high-throughput screening. PMID- 19219009 TI - Corrected structure of mirin, a small-molecule inhibitor of the Mre11-Rad50-Nbs1 complex. PMID- 19219012 TI - Membrane proteins take center stage in Frankfurt. AB - Biological membranes are dynamic frontiers whose molecules must delicately balance the needs for compartmentalization and communication, and the gap between the vital significance of transport and signaling through membranes and our poor understanding of the precise functionality of these processes is daunting. However, a recent conference highlighted promising progress in the field, particularly made possible by the increasing structural knowledge about membrane proteins. PMID- 19219011 TI - Building a new conceptual framework for receptor heteromers. AB - Receptor heteromers constitute a new area of research that is reshaping our thinking about biochemistry, cell biology, pharmacology and drug discovery. In this commentary, we recommend clear definitions that should facilitate both information exchange and research on this growing class of transmembrane signal transduction units and their complex properties. We also consider research questions underlying the proposed nomenclature, with recommendations for receptor heteromer identification in native tissues and their use as targets for drug development. PMID- 19219013 TI - Alkaloid biosynthesis takes root. PMID- 19219014 TI - Intracellular zinc irritates TRPA1. PMID- 19219015 TI - Ironing out a new siderophore synthesis strategy. PMID- 19219016 TI - A pulse at the heart of targeted therapy. PMID- 19219017 TI - Signal sequences get active. PMID- 19219022 TI - Phase 2 gene therapy trial of an anti-HIV ribozyme in autologous CD34+ cells. AB - Gene transfer has potential as a once-only treatment that reduces viral load, preserves the immune system and avoids lifetime highly active antiretroviral therapy. This study, which is to our knowledge the first randomized, double blind, placebo-controlled, phase 2 cell-delivered gene transfer clinical trial, was conducted in 74 HIV-1-infected adults who received a tat-vpr-specific anti HIV ribozyme (OZ1) or placebo delivered in autologous CD34+ hematopoietic progenitor cells. There were no OZ1-related adverse events. There was no statistically significant difference in viral load between the OZ1 and placebo group at the primary end point (average at weeks 47 and 48), but time-weighted areas under the curve from weeks 40-48 and 40-100 were significantly lower in the OZ1 group. Throughout the 100 weeks, CD4+ lymphocyte counts were higher in the OZ1 group. This study indicates that cell-delivered gene transfer is safe and biologically active in individuals with HIV and can be developed as a conventional therapeutic product. PMID- 19219023 TI - Sensitive in vivo imaging of T cells using a membrane-bound Gaussia princeps luciferase. AB - We developed a new approach to bioluminescent T cell imaging using a membrane anchored form of the Gaussia luciferase (GLuc) enzyme, termed extGLuc, which we could stably express in both mouse and human primary T cells. In vitro, extGLuc+ cells emitted significantly higher bioluminescent signal when compared to cells expressing GLuc, Renilla luciferase (RLuc) or membrane-anchored RLuc (extRLuc). In vivo, mouse extGLuc+ T cells showed higher bioluminescent signal when compared to GLuc+ and RLuc+ T cells. Application of this imaging approach to human T cells genetically modified to express tumor-specific chimeric antigen receptors (CARs) enabled us to show in vivo CAR-mediated T cell accumulation in tumor, T cell persistence over time and concomitant imaging of T cells and tumor cells modified to express firefly luciferase. This sensitive imaging technology has application to many in vivo cell-based studies in a wide array of mouse models. PMID- 19219024 TI - Effector memory T cell responses are associated with protection of rhesus monkeys from mucosal simian immunodeficiency virus challenge. AB - The rapid onset of massive, systemic viral replication during primary HIV or simian immunodeficiency virus (SIV) infection and the immune evasion capabilities of these viruses pose fundamental problems for vaccines that depend upon initial viral replication to stimulate effector T cell expansion and differentiation. We hypothesized that vaccines designed to maintain differentiated effector memory T cell (TEM cell) responses at viral entry sites might improve efficacy by impairing viral replication at its earliest stage, and we have therefore developed SIV protein-encoding vectors based on rhesus cytomegalovirus (RhCMV), the prototypical inducer of life-long TEM cell responses. RhCMV vectors expressing SIV Gag, Rev-Tat-Nef and Env persistently infected rhesus macaques, regardless of preexisting RhCMV immunity, and primed and maintained robust, SIV specific CD4+ and CD8+ TEM cell responses (characterized by coordinate tumor necrosis factor, interferon-gamma and macrophage inflammatory protein-1beta expression, cytotoxic degranulation and accumulation at extralymphoid sites) in the absence of neutralizing antibodies. Compared to control rhesus macaques, these vaccinated rhesus macaques showed increased resistance to acquisition of progressive SIVmac239 infection upon repeated limiting-dose intrarectal challenge, including four macaques who controlled rectal mucosal infection without progressive systemic dissemination. These data suggest a new paradigm for AIDS vaccine development--vaccines capable of generating and maintaining HIV specific TEM cells might decrease the incidence of HIV acquisition after sexual exposure. PMID- 19219025 TI - Phylogenetic biome conservatism on a global scale. AB - How and why organisms are distributed as they are has long intrigued evolutionary biologists. The tendency for species to retain their ancestral ecology has been demonstrated in distributions on local and regional scales, but the extent of ecological conservatism over tens of millions of years and across continents has not been assessed. Here we show that biome stasis at speciation has outweighed biome shifts by a ratio of more than 25:1, by inferring ancestral biomes for an ecologically diverse sample of more than 11,000 plant species from around the Southern Hemisphere. Stasis was also prevalent in transocean colonizations. Availability of a suitable biome could have substantially influenced which lineages establish on more than one landmass, in addition to the influence of the rarity of the dispersal events themselves. Conversely, the taxonomic composition of biomes has probably been strongly influenced by the rarity of species' transitions between biomes. This study has implications for the future because if clades have inherently limited capacity to shift biomes, then their evolutionary potential could be strongly compromised by biome contraction as climate changes. PMID- 19219026 TI - c-Myc suppression of miR-23a/b enhances mitochondrial glutaminase expression and glutamine metabolism. AB - Altered glucose metabolism in cancer cells is termed the Warburg effect, which describes the propensity of most cancer cells to take up glucose avidly and convert it primarily to lactate, despite available oxygen. Notwithstanding the renewed interest in the Warburg effect, cancer cells also depend on continued mitochondrial function for metabolism, specifically glutaminolysis that catabolizes glutamine to generate ATP and lactate. Glutamine, which is highly transported into proliferating cells, is a major source of energy and nitrogen for biosynthesis, and a carbon substrate for anabolic processes in cancer cells, but the regulation of glutamine metabolism is not well understood. Here we report that the c-Myc (hereafter referred to as Myc) oncogenic transcription factor, which is known to regulate microRNAs and stimulate cell proliferation, transcriptionally represses miR-23a and miR-23b, resulting in greater expression of their target protein, mitochondrial glutaminase, in human P-493 B lymphoma cells and PC3 prostate cancer cells. This leads to upregulation of glutamine catabolism. Glutaminase converts glutamine to glutamate, which is further catabolized through the tricarboxylic acid cycle for the production of ATP or serves as substrate for glutathione synthesis. The unique means by which Myc regulates glutaminase uncovers a previously unsuspected link between Myc regulation of miRNAs, glutamine metabolism, and energy and reactive oxygen species homeostasis. PMID- 19219027 TI - Identification of a dendritic cell receptor that couples sensing of necrosis to immunity. AB - Injury or impaired clearance of apoptotic cells leads to the pathological accumulation of necrotic corpses, which induce an inflammatory response that initiates tissue repair. In addition, antigens present in necrotic cells can sometimes provoke a specific immune response and it has been argued that necrosis could explain adaptive immunity in seemingly infection-free situations, such as after allograft transplantation or in spontaneous and therapy-induced tumour rejection. In the mouse, the CD8alpha+ subset of dendritic cells phagocytoses dead cell remnants and cross-primes CD8+ T cells against cell-associated antigens. Here we show that CD8alpha+ dendritic cells use CLEC9A (also known as DNGR-1), a recently-characterized C-type lectin, to recognize a preformed signal that is exposed on necrotic cells. Loss or blockade of CLEC9A does not impair the uptake of necrotic cell material by CD8+ dendritic cells, but specifically reduces cross-presentation of dead-cell-associated antigens in vitro and decreases the immunogenicity of necrotic cells in vivo. The function of CLEC9A requires a key tyrosine residue in its intracellular tail that allows the recruitment and activation of the tyrosine kinase SYK, which is also essential for cross-presentation of dead-cell-associated antigens. Thus, CLEC9A functions as a SYK-coupled C-type lectin receptor to mediate sensing of necrosis by the principal dendritic-cell subset involved in regulating cross-priming to cell associated antigens. PMID- 19219028 TI - Gene silencing by synthetic U1 adaptors. AB - We describe a gene silencing method that employs a mechanism of action distinct from those of antisense and RNA interference. U1 Adaptors are bifunctional oligonucleotides with a 'target domain' complementary to a site in the target gene's terminal exon and a 'U1 domain' that binds to the U1 small nuclear RNA component of the U1 small nuclear ribonucleoprotein (U1 snRNP) splicing factor. Tethering of U1 snRNP to the target pre-mRNA inhibits poly(A)-tail addition, causing degradation of that RNA species in the nucleus. U1 Adaptors can inhibit both endogenous and reporter genes in a sequence-specific manner. Comparison of U1 Adaptors with small interfering RNA (siRNA) using a genome-wide microarray analysis indicates that U1 Adaptors have limited off-target effects and no detectable adverse effects on splicing. Further, targeting the same gene either with multiple U1 Adaptors or with a U1 Adaptor and siRNA strongly enhances gene silencing. PMID- 19219029 TI - Room-temperature defect-engineered spin filter based on a non-magnetic semiconductor. AB - Generating, manipulating and detecting electron spin polarization and coherence at room temperature is at the heart of future spintronics and spin-based quantum information technology. Spin filtering, which is a key issue for spintronic applications, has been demonstrated by using ferromagnetic metals, diluted magnetic semiconductors, quantum point contacts, quantum dots, carbon nanotubes, multiferroics and so on. This filtering effect was so far restricted to a limited efficiency and primarily at low temperatures or under a magnetic field. Here, we provide direct and unambiguous experimental proof that an electron-spin-polarized defect, such as a Ga(i) self-interstitial in dilute nitride GaNAs, can effectively deplete conduction electrons with an opposite spin orientation and can thus turn the non-magnetic semiconductor into an efficient spin filter operating at room temperature and zero magnetic field. This work shows the potential of such defect-engineered, switchable spin filters as an attractive alternative to generate, amplify and detect electron spin polarization at room temperature without a magnetic material or external magnetic fields. PMID- 19219030 TI - Giant superconductivity-induced modulation of the ferromagnetic magnetization in a cuprate-manganite superlattice. AB - Artificial multilayers offer unique opportunities for combining materials with antagonistic orders such as superconductivity and ferromagnetism and thus to realize novel quantum states. In particular, oxide multilayers enable the utilization of the high superconducting transition temperature of the cuprates and the versatile magnetic properties of the colossal-magnetoresistance manganites. However, apart from exploratory work, the in-depth investigation of their unusual properties has only just begun. Here we present neutron reflectometry measurements of a [Y(0.6)Pr(0.4)Ba(2)Cu(3)O(7) (10 nm)/La(2/3)Ca(1/3)MnO(3) (10 nm)](10) superlattice, which reveal a surprisingly large superconductivity-induced modulation of the vertical ferromagnetic magnetization profile. Most surprisingly, this modulation seems to involve the density rather than the orientation of the magnetization and is highly susceptible to the strain, which is transmitted from the SrTiO(3) substrate. We outline a possible explanation of this unusual superconductivity-induced phenomenon in terms of a phase separation between ferromagnetic and non ferromagnetic nanodomains in the La(2/3)Ca(1/3)MnO(3) layers. PMID- 19219031 TI - Mesoporous germanium-rich chalcogenido frameworks with highly polarizable surfaces and relevance to gas separation. AB - Mesoporous materials with tunable non-oxidic framework compositions can exhibit new kinds of functionality including internal surfaces with high polarizability. As the chemical and physical characteristics of the framework components can induce useful catalytic, absorption and optoelectronic features, the mesoporous structure can promote fast mass diffusion kinetics and size-selective transport of guest molecules. So far, synthetic efforts have resulted in mesoporous metal chalcogenides on using structure-directing moulds of soft or hard templates. These include ordered mesoporous II-VI semiconductors (such as CdS (refs 2,3), ZnS (ref. 4) and CdTe (ref. 5)). Recently, template-free synthetic routes for high-surface-area chalcogenide aerogels have been reported. Here, we describe a novel kind of porous materials based on germanium-rich chalcogenide networks and 'soft' highly polarizable surfaces. We demonstrate that these materials can exhibit excellent selectivity for separating hydrogen from carbon dioxide and methane. These highly polarizable mesoporous structures have important implications for membrane-based gas separation process technologies including hydrogen purification. PMID- 19219032 TI - The influence of edge structure on the electronic properties of graphene quantum dots and nanoribbons. AB - Graphene shows promise as a future material for nanoelectronics owing to its compatibility with industry-standard lithographic processing, electron mobilities up to 150 times greater than Si and a thermal conductivity twice that of diamond. The electronic structure of graphene nanoribbons (GNRs) and quantum dots (GQDs) has been predicted to depend sensitively on the crystallographic orientation of their edges; however, the influence of edge structure has not been verified experimentally. Here, we use tunnelling spectroscopy to show that the electronic structure of GNRs and GQDs with 2-20 nm lateral dimensions varies on the basis of the graphene edge lattice symmetry. Predominantly zigzag-edge GQDs with 7-8 nm average dimensions are metallic owing to the presence of zigzag edge states. GNRs with a higher fraction of zigzag edges exhibit a smaller energy gap than a predominantly armchair-edge ribbon of similar width, and the magnitudes of the measured GNR energy gaps agree with recent theoretical calculations. PMID- 19219033 TI - Automated light-based mapping of motor cortex by photoactivation of channelrhodopsin-2 transgenic mice. AB - Traditionally, mapping the motor cortex requires electrodes to stimulate the brain and define motor output pathways. Although effective, electrode-based methods are labor-intensive, potentially damaging to the cortex and can have off target effects. As an alternative method of motor mapping, we photostimulated transgenic mice expressing the light-sensitive ion channel channelrhodopsin-2 in predominantly layer-5 output cortical neurons. We report that optical stimulation of these neurons in vivo using a stage scanning laser system resulted in muscle excitation within 10-20 ms, which can be recorded using implanted electromyogram electrodes or by a noninvasive motion sensor. This approach allowed us to make highly reproducible automated maps of the mouse forelimb and hindlimb motor cortex much faster than with previous methods. We anticipate that the approach will facilitate the study of changes in the location and properties of motor maps after skilled training or damage to the nervous system. PMID- 19219034 TI - The yeast global transcriptional co-repressor protein Cyc8 can propagate as a prion. AB - Although many proteins can misfold into a self-seeding amyloid-like conformation, only six are known to be infectious, that is prions. The prions [PSI(+)], [PIN(+)], [URE3], [SWI(+)] and [HET-s] cause distinct heritable physiological changes in fungi, whereas PrP(Sc) causes infectious encephalopathies in mammals. It is unknown whether 'protein-only' inheritance is limited to these exceptional cases or whether it represents a widespread mechanism of epigenetic control. Towards this goal, we now describe a new prion formed by the Cyc8 (Ssn6) protein of Saccharomyces cerevisiae. Analogously to other yeast prions, transient overproduction of a glutamine-rich region of Cyc8 induced a heritable dominant cyc8(-) phenotype that is transmitted cytoplasmically and is dependent on the chaperone Hsp104 and the continued presence of the Cyc8 protein. The evolutionarily conserved Cyc8-Tup1 global transcriptional repressor complex forms one of the largest gene regulatory circuits, controlling the expression of more than 7% of yeast genes. Our finding that Cyc8 can propagate as a prion, together with a recent report that Swi1 of the Swi-Snf global transcriptional regulatory complex also has a prion form, shows that prionization can lead to mass activation or repression of yeast genes and is suggestive of a link between the epigenetic phenomena of chromatin remodelling and prion formation. PMID- 19219035 TI - Oct1 loss of function induces a coordinate metabolic shift that opposes tumorigenicity. AB - Cancer cells frequently undergo a shift from oxidative to glycolytic metabolism. Although there is interest in targeting metabolism as a form of cancer therapy, this area still remains in its infancy. Using cells, embryos and adult animals, we show here that loss of the widely expressed transcription factor Oct1 induces a coordinated metabolic shift: mitochondrial activity and amino acid oxidation are increased, while glucose metabolism is reduced. Altered expression of direct Oct1 targets encoding metabolic regulators provides a mechanistic underpinning to these results. We show that these metabolic changes directly oppose tumorigenicity. Collectively, our findings show that Oct1, the genes it regulates and the pathways these genes affect could be used as targets for new modes of cancer therapy. PMID- 19219036 TI - NuMA-related LIN-5, ASPM-1, calmodulin and dynein promote meiotic spindle rotation independently of cortical LIN-5/GPR/Galpha. AB - The spindle apparatus dictates the plane of cell cleavage, which is critical in the choice between symmetric or asymmetric division. Spindle positioning is controlled by an evolutionarily conserved pathway, which involves LIN-5/GPR 1/2/Galpha in Caenorhabditis elegans, Mud/Pins/Galpha in Drosophila and NuMA/LGN/Galpha in humans. GPR-1/2 and Galpha localize LIN-5 to the cell cortex, which engages dynein and controls the cleavage plane during early mitotic divisions in C. elegans. Here we identify ASPM-1 (abnormal spindle-like, microcephaly-associated) as a novel LIN-5 binding partner. ASPM-1, together with calmodulin (CMD-1), promotes meiotic spindle organization and the accumulation of LIN-5 at meiotic and mitotic spindle poles. Spindle rotation during maternal meiosis is independent of GPR-1/2 and Galpha, yet requires LIN-5, ASPM-1, CMD-1 and dynein. Our data support the existence of two distinct LIN-5 complexes that determine localized dynein function: LIN-5/GPR-1/2/Galpha at the cortex, and LIN 5/ASPM-1/CMD-1 at spindle poles. These functional interactions may be conserved in mammals, with implications for primary microcephaly. PMID- 19219037 TI - An anatomic gene expression atlas of the adult mouse brain. AB - Studying gene expression provides a powerful means of understanding structure function relationships in the nervous system. The availability of genome-scale in situ hybridization datasets enables new possibilities for understanding brain organization based on gene expression patterns. The Anatomic Gene Expression Atlas (AGEA) is a new relational atlas revealing the genetic architecture of the adult C57Bl/6J mouse brain based on spatial correlations across expression data for thousands of genes in the Allen Brain Atlas (ABA). The AGEA includes three discovery tools for examining neuroanatomical relationships and boundaries: (1) three-dimensional expression-based correlation maps, (2) a hierarchical transcriptome-based parcellation of the brain and (3) a facility to retrieve from the ABA specific genes showing enriched expression in local correlated domains. The utility of this atlas is illustrated by analysis of genetic organization in the thalamus, striatum and cerebral cortex. The AGEA is a publicly accessible online computational tool integrated with the ABA (http://mouse.brain map.org/agea). PMID- 19219038 TI - Beyond extinction: erasing human fear responses and preventing the return of fear. AB - Animal studies have shown that fear memories can change when recalled, a process referred to as reconsolidation. We found that oral administration of the beta adrenergic receptor antagonist propranolol before memory reactivation in humans erased the behavioral expression of the fear memory 24 h later and prevented the return of fear. Disrupting the reconsolidation of fear memory opens up new avenues for providing a long-term cure for patients with emotional disorders. PMID- 19219039 TI - Role of the synaptic ribbon in transmitting the cone light response. AB - Cone photoreceptors distinguish small changes in light intensity while operating over a wide dynamic range. The cone synapse encodes intensity by modulating tonic neurotransmitter release, but precise encoding is limited by the quantal nature of synaptic vesicle exocytosis. Cones possess synaptic ribbons, structures that are thought to accelerate the delivery of vesicles for tonic release. Here we show that the synaptic ribbon actually constrains vesicle delivery, resulting in a maintained state of synaptic depression in darkness. Electron microscopy of cones from the lizard Anolis segrei revealed that depression is caused by the depletion of vesicles on the ribbon, indicating that resupply, not fusion, is the rate-limiting step that controls release. Responses from postsynaptic retinal neurons from the salamander Ambystoma tigrinum showed that the ribbon behaves like a capacitor, charging with vesicles in light and discharging in a phasic burst at light offset. Phasic release extends the operating range of the cone synapse to more accurately encode changes in light intensity, accentuating features that are salient to photopic vision. PMID- 19219040 TI - Reinforcement learning in populations of spiking neurons. AB - Population coding is widely regarded as an important mechanism for achieving reliable behavioral responses despite neuronal variability. However, standard reinforcement learning slows down with increasing population size, as the global reward signal becomes less and less related to the performance of any single neuron. We found that learning speeds up with increasing population size if, in addition to global reward, feedback about the population response modulates synaptic plasticity. PMID- 19219041 TI - Association of common variants in NPPA and NPPB with circulating natriuretic peptides and blood pressure. AB - We examined the association of common variants at the NPPA-NPPB locus with circulating concentrations of the natriuretic peptides, which have blood pressure lowering properties. We genotyped SNPs at the NPPA-NPPB locus in 14,743 individuals of European ancestry, and identified associations of plasma atrial natriuretic peptide with rs5068 (P = 8 x 10(-70)), rs198358 (P = 8 x 10(-30)) and rs632793 (P = 2 x 10(-10)), and of plasma B-type natriuretic peptide with rs5068 (P = 3 x 10(-12)), rs198358 (P = 1 x 10(-25)) and rs632793 (P = 2 x 10(-68)). In 29,717 individuals, the alleles of rs5068 and rs198358 that showed association with increased circulating natriuretic peptide concentrations were also found to be associated with lower systolic (P = 2 x 10(-6) and 6 x 10(-5), respectively) and diastolic blood pressure (P = 1 x 10(-6) and 5 x 10(-5)), as well as reduced odds of hypertension (OR = 0.85, 95% CI = 0.79-0.92, P = 4 x 10(-5); OR = 0.90, 95% CI = 0.85-0.95, P = 2 x 10(-4), respectively). Common genetic variants at the NPPA-NPPB locus found to be associated with circulating natriuretic peptide concentrations contribute to interindividual variation in blood pressure and hypertension. PMID- 19219042 TI - Genome-wide association study identifies a new breast cancer susceptibility locus at 6q25.1. AB - We carried out a genome-wide association study among Chinese women to identify risk variants for breast cancer. After analyzing 607,728 SNPs in 1,505 cases and 1,522 controls, we selected 29 SNPs for a fast-track replication in an independent set of 1,554 cases and 1,576 controls. We further investigated four replicated loci in a third set of samples comprising 3,472 cases and 900 controls. SNP rs2046210 at 6q25.1, located upstream of the gene encoding estrogen receptor alpha (ESR1), showed strong and consistent association with breast cancer across all three stages. Adjusted odds ratio (95% CI) were 1.36 (1.24 1.49) and 1.59 (1.40-1.82), respectively, for genotypes A/G and A/A versus G/G (P for trend 2.0 x 10(-15)) in the pooled analysis of samples from all three stages. We also found a similar, albeit weaker, association in an independent study comprising 1,591 cases and 1,466 controls of European ancestry (P(trend) = 0.01). These results strongly implicate 6q25.1 as a susceptibility locus for breast cancer. PMID- 19219043 TI - A TARBP2 mutation in human cancer impairs microRNA processing and DICER1 function. AB - microRNAs (miRNAs) are small noncoding RNAs that regulate gene expression by targeting messenger RNA (mRNA) transcripts. Recently, a miRNA expression profile of human tumors has been characterized by an overall miRNA downregulation. Explanations for this observation include a failure of miRNA post-transcriptional regulation, transcriptional silencing associated with hypermethylation of CpG island promoters and miRNA transcriptional repression by oncogenic factors. Another possibility is that the enzymes and cofactors involved in miRNA processing pathways may themselves be targets of genetic disruption, further enhancing cellular transformation. However, no loss-of-function genetic alterations in the genes encoding these proteins have been reported. Here we have identified truncating mutations in TARBP2 (TAR RNA-binding protein 2), encoding an integral component of a DICER1-containing complex, in sporadic and hereditary carcinomas with microsatellite instability. The presence of TARBP2 frameshift mutations causes diminished TRBP protein expression and a defect in the processing of miRNAs. The reintroduction of TRBP in the deficient cells restores the efficient production of miRNAs and inhibits tumor growth. Most important, the TRBP impairment is associated with a destabilization of the DICER1 protein. These results provide, for a subset of human tumors, an explanation for the observed defects in the expression of mature miRNAs. PMID- 19219044 TI - FGF9 monomer-dimer equilibrium regulates extracellular matrix affinity and tissue diffusion. AB - The spontaneous dominant mouse mutant, Elbow knee synostosis (Eks), shows elbow and knee joint synosotsis, and premature fusion of cranial sutures. Here we identify a missense mutation in the Fgf9 gene that is responsible for the Eks mutation. Through investigation of the pathogenic mechanisms of joint and suture synostosis in Eks mice, we identify a key molecular mechanism that regulates FGF9 signaling in developing tissues. We show that the Eks mutation prevents homodimerization of the FGF9 protein and that monomeric FGF9 binds to heparin with a lower affinity than dimeric FGF9. These biochemical defects result in increased diffusion of the altered FGF9 protein (FGF9(Eks)) through developing tissues, leading to ectopic FGF9 signaling and repression of joint and suture development. We propose a mechanism in which the range of FGF9 signaling in developing tissues is limited by its ability to homodimerize and its affinity for extracellular matrix heparan sulfate proteoglycans. PMID- 19219045 TI - A Mek1-Mek2 heterodimer determines the strength and duration of the Erk signal. AB - Mek1 and Mek2 (also known as Map2k1 and Map2k2, respectively) are evolutionarily conserved, dual-specificity kinases that mediate Erk1 and Erk2 activation during adhesion and growth factor signaling. Here we describe a previously uncharacterized, unexpected role of Mek1 in downregulating Mek2-dependent Erk signaling. Mek1 mediates the regulation of Mek2 in the context of a previously undiscovered Mek1-Mek2 complex. The Mek heterodimer is negatively regulated by Erk-mediated phosphorylation of Mek1 on Thr292, a residue missing in Mek2. Disabling this Erk-proximal negative-feedback step stabilizes the phosphorylation of both Mek2 and Erk in cultured cells and in vivo in Mek1 knockout embryos and mice. Thus, in disagreement with the current perception of the pathway, the role of Mek1 and Mek2 in growth factor-induced Erk phosphorylation is not interchangeable. Our data establish Mek1 as the crucial modulator of Mek and Erk signaling and have potential implications for the role of Mek1 and Mek2 in tumorigenesis. PMID- 19219046 TI - The mRNA export protein DBP5 binds RNA and the cytoplasmic nucleoporin NUP214 in a mutually exclusive manner. AB - The DEAD-box protein DBP5 is essential for mRNA export in both yeast and humans. It binds RNA and is concentrated and locally activated at the cytoplasmic side of the nuclear pore complex. We have determined the crystal structures of human DBP5 bound to RNA and AMPPNP, and bound to the cytoplasmic nucleoporin NUP214. The structures reveal that binding of DBP5 to nucleic acid and to NUP214 is mutually exclusive. Using in vitro assays, we demonstrate that NUP214 decreases both the RNA binding and ATPase activities of DBP5. The interactions are mediated by conserved residues, implying a conserved recognition mechanism. These results suggest a framework for the consecutive steps leading to the release of mRNA at the final stages of nuclear export. More generally, they provide a paradigm for how binding of regulators can specifically inhibit DEAD-box proteins. PMID- 19219047 TI - Structural basis for G9a-like protein lysine methyltransferase inhibition by BIX 01294. AB - Histone lysine methylation is an important epigenetic mark that regulates gene expression and chromatin organization. G9a and G9a-like protein (GLP) are euchromatin-associated methyltransferases that repress transcription by methylating histone H3 Lys9. BIX-01294 was originally identified as a G9a inhibitor during a chemical library screen of small molecules and has previously been used in the generation of induced pluripotent stem cells. Here we present the crystal structure of the catalytic SET domain of GLP in complex with BIX 01294 and S-adenosyl-L-homocysteine. The inhibitor is bound in the substrate peptide groove at the location where the histone H3 residues N-terminal to the target lysine lie in the previously solved structure of the complex with histone peptide. The inhibitor resembles the bound conformation of histone H3 Lys4 to Arg8, and is positioned in place by residues specific for G9a and GLP through specific interactions. PMID- 19219049 TI - Analysis of the eukaryotic prenylome by isoprenoid affinity tagging. AB - Protein prenylation is a widespread phenomenon in eukaryotic cells that affects many important signaling molecules. We describe the structure-guided design of engineered protein prenyltransferases and their universal synthetic substrate, biotin-geranylpyrophosphate. These new tools allowed us to detect femtomolar amounts of prenylatable proteins in cells and organs and to identify their cognate protein prenyltransferases. Using this approach, we analyzed the in vivo effects of protein prenyltransferase inhibitors. Whereas some of the inhibitors displayed the expected activities, others lacked in vivo activity or targeted a broader spectrum of prenyltransferases than previously believed. To quantitate the in vivo effect of the prenylation inhibitors, we profiled biotin-geranyl tagged RabGTPases across the proteome by mass spectrometry. We also demonstrate that sites of active vesicular transport carry most of the RabGTPases. This approach enables a quantitative proteome-wide analysis of the regulation of protein prenylation and its modulation by therapeutic agents. PMID- 19219048 TI - Cyanobacterial photosystem II at 2.9-A resolution and the role of quinones, lipids, channels and chloride. AB - Photosystem II (PSII) is a large homodimeric protein-cofactor complex located in the photosynthetic thylakoid membrane that acts as light-driven water:plastoquinone oxidoreductase. The crystal structure of PSII from Thermosynechococcus elongatus at 2.9-A resolution allowed the unambiguous assignment of all 20 protein subunits and complete modeling of all 35 chlorophyll a molecules and 12 carotenoid molecules, 25 integral lipids and 1 chloride ion per monomer. The presence of a third plastoquinone Q(C) and a second plastoquinone-transfer channel, which were not observed before, suggests mechanisms for plastoquinol-plastoquinone exchange, and we calculated other possible water or dioxygen and proton channels. Putative oxygen positions obtained from a Xenon derivative indicate a role for lipids in oxygen diffusion to the cytoplasmic side of PSII. The chloride position suggests a role in proton transfer reactions because it is bound through a putative water molecule to the Mn(4)Ca cluster at a distance of 6.5 A and is close to two possible proton channels. PMID- 19219050 TI - Plasmodium genomics: latest milestone. AB - Our knowledge on comparative genomics of the malaria parasites has advanced a step forward with the publication of the genomes of two primate-infecting malaria parasites: Plasmodium vivax and Plasmodium knowlesi. Even though the genomes of these organisms are the fifth and sixth Plasmodium genomes to be sequenced, respectively, both have revealed previously unknown features, which are discussed in this month's Genome Watch. PMID- 19219052 TI - Under pressure: investigating the biology of plant infection by Magnaporthe oryzae. AB - The filamentous fungus Magnaporthe oryzae causes rice blast, the most serious disease of cultivated rice. Cellular differentiation of M. oryzae forms an infection structure called the appressorium, which generates enormous cellular turgor that is sufficient to rupture the plant cuticle. Here, we show how functional genomics approaches are providing new insight into the genetic control of plant infection by M. oryzae. We also look ahead to the key questions that need to be addressed to provide a better understanding of the molecular processes that lead to plant disease and the prospects for sustainable control of rice blast. PMID- 19219053 TI - Systems biology: Functional analysis of natural microbial consortia using community proteomics. AB - We know very little about the metabolic functioning and evolutionary dynamics of microbial communities. Recent advances in comprehensive, sequencing-based methods, however, are laying a molecular foundation for new insights into how microbial communities shape the Earth's biosphere. Here we explore the convergence of microbial ecology, genomics, biological mass spectrometry and informatics that form the new field of microbial community proteogenomics. We discuss the first applications of proteogenomics and its potential for studying the physiology, ecology and evolution of microbial populations and communities. PMID- 19219056 TI - Cell death mechanisms: life in the balance. PMID- 19219054 TI - Antimicrobial peptides: linking partition, activity and high membrane-bound concentrations. AB - An increasing amount of information on the action of antimicrobial peptides (AMPs) at the molecular level has not yet been translated into a comprehensive understanding of effects in bacteria. Although some biophysical attributes of AMPs have been correlated with macroscopic features, the physiological relevance of other properties has not yet been addressed. Pertinent and surprising conclusions have therefore been left unstated. Strong membrane-binding and micromolar therapeutic concentrations of AMPs indicate that membrane-bound concentrations may be reached that are higher than intuitively expected, triggering disruptive effects on bacteria. PMID- 19219057 TI - Clinical observation of loupe-assisted intussusception vasoepididymostomy in the treatment of obstructive azoospermia (analysis of 49 case reports). AB - To evaluate the clinical outcomes of loupe-assisted intussusception vasoepididymostomy (VE) in the treatment of epididymal obstructive azoospermia (EOA), we retrospectively analyzed data from 49 patients with EOA who underwent two-suture longitudinal intussusception vasoepididymostomy (LIVE) between 2000 and 2007. The data included the surgical method, postoperative motile sperm count per ejaculation, percentage of progressive motile sperm and patency and pregnancy outcomes. There were a total of 49 men undergoing scrotal exploration, and epididymal obstruction was found in all cases. Bilateral or unilateral anastomoses were performed in 40 and 6 men, respectively. The postoperative courses of 42 patients were followed up for more than 6 months, and the courses of 38 patients were followed up for more than 1 year. The overall patency and pregnancy rates were 71.4% and 26.3%, respectively. Moreover, progressive motile sperm was more frequently present in those patients who had undergone anastomosis at cauda than at corpus or caput. Pregnancy was achieved only in those patients who had undergone anastomosis at least on one side of the cauda epididymis. We think that the loupe-assisted method, with a lower overall cost and a simplified surgical procedure, can achieve satisfactory patency outcomes and pregnancy results. Data from this paper also suggest that paternity outcomes occur more frequently after anastomoses at cauda than at corpus or caput. PMID- 19219059 TI - Genetic obesity affects neural ketone body utilization in the rat brain. AB - Obesity causes various physiological disorders between the central nervous system and peripheral tissues. Ketone bodies have a neuro-protective role and are strongly affected by obesity-related metabolic disorders. To clarify the effects of obesity on ketone body utilization in brain, we examined the mRNA localization of acetoacetyl-CoA synthetase (AACS), which activates ketone bodies for the synthesis of fatty acid and cholesterol, in various brain regions of Zucker fatty rats by in situ hybridization. The AACS mRNA level was increased in the paraventricular thalamic nucleus (PVT) but not affected in the cerebrum and hippocampus in Zucker fatty rats. In contrast, the AACS mRNA level was reduced in the arcuate hypothalamic nucleus (Arc) and ventromedial hypothalamic nucleus (VMH) in the hypothalamus. Succinyl-CoA:3-oxoacid CoA-transferase (SCOT) mRNA level was decreased only in the PVT but not affected in the Arc and VMH. These data raise the possibility that AACS is regulated by the leptin signaling pathway in the hypothalamus but not in the PVT. As AACS was expressed in neural-like cells, ketone bodies are assumed to be utilized for the synthesis of lipidic substances and to cause metabolic disorders in the nervous system. PMID- 19219058 TI - The antibody against a nuclear autoantigenic sperm protein can result in reproductive failure. AB - To study whether the antibody against the testis form of the nuclear autoantigenic sperm protein (tNASP) could result in reproductive failure, we successfully cloned and expressed a 339-bp cDNA fragment of mouse tNASP (mtNASP). Using mouse as a model, recombinant mtNASP (rmtNASP) and a synthetic peptide, human tNASP(393-408) (htNASP(393-408)), were investigated for their antifertility effect. Active immunization with rmtNASP or the synthesized peptide raised high antibody titers in the immunized mice. Sperm-egg binding and fusion assay were carried out in 8-10-week-old BALB/c mice. Sperm-egg binding and in vitro fertilization of mouse oocytes were inhibited by co-incubation of zona-free mouse oocytes with capacitated mouse spermatozoa in the presence of varying concentrations of the antisera against rmtNASP. There was a significant antifertility effect in animals immunized with rmtNASP or the synthesized peptide. The effect on fertility in the mice immunized with the synthesized peptide was reversible. Our data indicate that active immunization with rmtNASP antigen may induce a strong antibody response that causes an inhibition of fertility. PMID- 19219060 TI - Obesity is associated with altered lung function independently of physical activity and fitness. AB - Measures of obesity, especially central adiposity, have been associated with reduced lung function. However, previous studies may have been affected by confounding by physical activity and fitness. This study aimed to examine the relationship among body fatness, fat distribution, and lung function, adjusted for physical activity energy expenditure (PAEE) and aerobic fitness (VO(2)max), in a cohort of British white adults with a family history of type 2 diabetes. A total of 320 adults (mean age 40.4 +/- 6.0 years) attended for anthropometric and VO(2)max testing, and had ambulatory heart rate monitoring for 4 days to determine PAEE. Spirometry was used to measure forced expiratory volume in 1 s (FEV(1)) and forced vital capacity (FVC). The tests were repeated 12 months later, and a cross-sectional analysis using linear regression with repeated measures was performed. Measures of obesity (BMI, waist circumference (WC), fat mass (FM), body fat percentage (BF%)) were associated with lower lung function in men and women (P < 0.01), while waist-to-hip ratio (WHR) was associated with lower lung function in men only (P < 0.001). Associations remained after adjusting for age, smoking status, height, PAEE, and VO(2)max. The estimated difference in mean FEV(1) and FVC per unit increase in the exposure measures were consistently stronger in men compared to women (P for interaction <0.001). Obesity is inversely associated with lung function in adults, but central fat distribution appears to have a stronger relationship with respiratory mechanics in men than in women. These associations were independent of the degree of physical activity and aerobic fitness in this cohort. PMID- 19219061 TI - Fat depot-specific impact of visceral obesity on adipocyte adiponectin release in women. AB - Our objective was to examine omental and subcutaneous adipocyte adiponectin release in women. We tested the hypothesis that adiponectin release would be reduced to a greater extent in omental than in subcutaneous adipocytes of women with visceral obesity. Omental and subcutaneous adipose tissue samples were obtained from 52 women undergoing abdominal hysterectomies (age: 47.1 +/- 4.8 years; BMI: 26.7 +/- 4.7 kg/m(2)). Adipocytes were isolated and their adiponectin release in the medium was measured over 2 h. Measures of body fat accumulation and distribution were obtained using dual-energy X-ray absorptiometry and computed tomography, respectively. Adiponectin release by omental and subcutaneous adipocytes was similar in lean individuals; however, in subsamples of obese or visceral obese women, adiponectin release by omental adipocytes was significantly reduced while that of subcutaneous adipocytes was not affected. Omental adipocyte adiponectin release was significantly and negatively correlated with total body fat mass (r = -0.47, P < 0.01), visceral adipose tissue area (r = -0.50, P < 0.01), omental adipocyte diameter (r = -0.43, P < 0.01), triglyceride levels (r = -0.32, P < or = 0.05), cholesterol/high-density lipoprotein (HDL) cholesterol (r = -0.31, P < or = 0.05), fasting glucose (r = -0.39, P < or = 0.01), fasting insulin (r = -0.36, P < or = 0.05), homeostasis model assessment index (r = -0.39, P < or = 0.01), and positively associated with HDL-cholesterol concentrations (r = 0.33, P < or = 0.05). Adiponectin release from subcutaneous cells was not associated with any measure of adiposity, lipid profile, or glucose homeostasis. In conclusion, compared to subcutaneous adipocyte adiponectin release, omental adipocyte adiponectin release is reduced to a greater extent in visceral obese women and better predicts obesity-associated metabolic abnormalities. PMID- 19219062 TI - Loss of total and visceral adipose tissue mass predicts decreases in oxidative stress after weight-loss surgery. AB - It is not known whether there are mechanisms linking adipose tissue mass and increased oxidative stress in obesity. This study investigated associations between decreasing general and abdominal fat depots and oxidative stress during weight loss. Subjects were severely obese women who were measured serially at baseline and at 1, 6 (n = 30), and 24 months (n = 18) after bariatric surgery. Total fat mass (FAT) and volumes of visceral (VAT) and subcutaneous abdominal adipose tissue (SAT) were related to plasma concentrations of derivatives of reactive oxidative metabolites (dROMS), a measure of lipid peroxides and oxidative stress. After intervention, BMI significantly decreased, from 47.7 +/- 0.8 kg/m(2) to 43.3 +/- 0.8 kg/m(2) (1 month), 35.2 +/- 0.8 kg/m(2) (6 months), and 30.2 +/- 1.2 kg/m(2) (24 months). Plasma dROMS also significantly deceased over time. At baseline, VAT (r = 0.46), FAT (r = 0.42), and BMI (r = 0.37) correlated with 6-month decreases in dROMS. Similarly, at 1 month, VAT (r = 0.43) and FAT (r = 0.41) correlated with 6-month decreases in dROMS. Multiple regression analysis showed that relationships between VAT and dROMS were significant after adjusting for FAT mass. Increased plasma dROMS at baseline were correlated with decreased concentrations of high-density lipoprotein (HDL) at 1 and 6 months after surgery (r = -0.38 and -0.42). This study found longitudinal associations between general, and more specifically intra-abdominal adiposity, and systemic lipid peroxides, suggesting that adipose tissue mass contributes to oxidative stress. PMID- 19219063 TI - The metabolic syndrome and behavioral correlates in obese patients with binge eating disorder. AB - This study examined the frequency of the metabolic syndrome (MetSyn) and explored behavioral eating- and weight-related correlates in obese patients with binge eating disorder (BED). Ninety-three treatment-seeking obese BED patients (22 men and 71 women) with and without the MetSyn were compared on demographic features and a number of current and historical eating and weight variables. Sixty percent of the obese patients with BED met criteria for the MetSyn, with men and whites having significantly higher rates than women and African Americans, respectively. Patients with vs. without coexisting MetSyn did not differ significantly in self reported frequency of binge eating or severity of eating disorder psychopathology. Multivariate hierarchical logistic regression analysis revealed that, after controlling for gender, ethnicity, and BMI, fewer episodes of weight cycling and regular meal skipping were significant predictors of the MetSyn. These findings suggest that lifestyle behaviors including weight loss attempts and regular meal consumption may be potential targets for prevention and/or treatment of the MetSyn in obese patients with BED. PMID- 19219064 TI - 5-HT(1A) activation counteracts cardiovascular but not hypophagic effects of sibutramine in rats. AB - The noradrenaline (NA) and serotonin reuptake inhibitor, sibutramine, gives effective weight loss, but full efficacy cannot be attained at approved doses due to cardiovascular side effects. We assessed in rats the contributions of NA and serotonin transporters to sibutramine's hypophagic and cardiovascular effects, and whether selective 5-hydroxytryptamine (5-HT(1A)) receptor activation could counteract the latter without affecting the former. Food intake was assessed in freely feeding rats and cardiovascular parameters in conscious telemetered rats. Ex vivo radioligand binding was used to estimate brain monoamine transporter occupancy. Sibutramine (1-10 mg/kg p.o.) dose-dependently reduced food intake; however, 10 mg/kg p.o. markedly elevated blood pressure and heart rate. Sibutramine gave greater occupancy of NA than serotonin reuptake sites. Coadministration of the selective 5-HT(1A) agonist F-11440 (2.5 mg/kg p.o.) attenuated sibutramine-induced hypertension and tachycardia without altering its food intake effects. The selective NA reuptake inhibitors, nisoxetine or reboxetine, did not alter food intake alone, but each reduced food intake when combined with F-11440. These results suggest that sibutramine-induced hypophagic and cardiovascular effects are largely due to increased brain synaptic NA via NA reuptake inhibition, and that 5-HT(1A) activation can counter the undesirable cardiovascular effects resulting from increased sympathetic activity. Selective NA reuptake inhibitors did not reduce food intake alone but did when combined with 5-HT(1A) activation. Hence increased synaptic serotonin, via serotonin reuptake inhibition or 5-HT(1A) activation, together with increased NA, would appear to produce hypophagia. Thus weight loss with minimal cardiovascular risk could be achieved by 5-HT(1A) activation combined with NA transporter blockade. PMID- 19219065 TI - BMI and risk factors for suicide: why is BMI inversely related to suicide? AB - Higher BMI has been associated with a lower risk of suicide in large prospective studies, but the mechanisms for this link require elucidation. In the 2002 and 2004 iterations of the Behavioral Risk Factor Surveillance System (BRFSS), a large, population-based telephone survey of US adults conducted by the Centers for Disease Control and Prevention, participants reported their height, weight, and several potential risk factors for suicide, including alcohol use, mental health, marital status, firearm ownership, and risk-taking behaviors. We assessed whether BMI was associated with these risk factors among 224,247 eligible respondents in 2002 and 275,194 in 2004 after sample-weighted adjustment for age, race, region, smoking, and education. Alcohol-related risk factors tended to be lower with heavier BMI among women, while firearm-related risk factors tended to be higher with heavier BMI among men. Heavier BMI also tended to be associated with unmarried status and poor mental health, especially among women, and with infrequent seat belt use in men and women. No potential risk factors were consistently inversely associated with BMI in both sexes and years. In summary, in these samples of the US population, conventional risk factors for suicide were inconsistently associated with BMI, making them unlikely mediators for the observed relationship of BMI with lower risk of suicide. In some cases, risk factors were actually greater with heavier BMI. Further study of the relationship of BMI and suicide may yield novel modifiable risk factors that could cause or prevent this important cause of death. PMID- 19219066 TI - ATF3 and p15PAF are novel gatekeepers of genomic integrity upon UV stress. AB - After genotoxic stress, normal cells trigger DNA repair or, if unable to repair, undergo apoptosis to eradicate the cells that bear the risk of becoming tumorigenic. Here we show that repression of the transcription factor, activating transcription factor 3 (ATF3), after ultraviolet (UV)-mediated genotoxic stress impairs the DNA repair process. We provide evidence that ATF3 directly regulates the proliferating cell nuclear antigen (PCNA)-associated factor KIAA0101/p15(PAF). We further show that the expressions of ATF3 and p15(PAF) is sufficient to trigger the DNA repair machinery, and that attenuation of their expression alters DNA repair mechanisms. We show that the expression of p15(PAF) compensates for a lack of ATF3 expression, thereby constituting a major effector of ATF3 in the DNA repair process. In addition, we provide evidence that p15(PAF) expression is required for the correct function of PCNA during DNA repair, as prevention of their interaction significantly alters DNA repair mechanisms. Finally, defective DNA repair, because of the downregulation of p15(PAF) expression, rendered the cells more sensitive to UV-induced cell death. Therefore, our results suggest ATF3 and p15(PAF) as novel gatekeepers of genomic integrity after UV exposure. PMID- 19219067 TI - Horizontal transfer of tumor DNA to endothelial cells in vivo. AB - Tumor endothelial cells have long been regarded as genomically stable and therefore less likely to develop resistance to antiangiogenic therapies. However, recent findings have challenged this notion. We have shown that DNA can be transferred between cells through phagocytosis of apoptotic bodies by adjacent viable cells. Propagation of the ingested DNA is prevented by the activation of the p53-p21 pathway. In this study, we examined whether concomitant transfer of tumor DNA with genes that inactivate the p53 pathway could overcome the barrier to tumor DNA propagation. Our results demonstrate that fibroblasts and endothelial cells are capable of acquiring and replicating tumor DNA when the apoptotic tumor cells contain the SV40 large T antigen. Analysis of the tumor stroma of xenotransplanted tumors in severe combined immunodeficient mice revealed that a sub-population of the endothelial cells contained tumor DNA. These cells maintained the ability to form functional vessels in an in vivo assay and concurrently express tumor-encoded and endothelial-specific genes. PMID- 19219068 TI - Glial cell missing-1 transcription factor is required for the differentiation of the human trophoblast. AB - Mammalian placentation is a highly regulated process and is dependent on the proper development of specific trophoblast cell lineages. The two major types of trophoblast, villous and extravillous, show mitotic arrest during differentiation. In mice, the transcription factor, glial cell missing-1 (Gcm1), blocks mitosis and is required for syncytiotrophoblast formation and morphogenesis of the labyrinth, the murine equivalent of the villous placenta. The human homolog GCM1 has an analogous expression pattern, but its function is presently unknown. We studied GCM1 function in the human-derived BeWo choriocarcinoma cell line and in first trimester human placental villous and extravillous explants. The GCM1 expression was either inhibited by siRNA and antisense oligonucleotides methods or upregulated by forskolin treatment. Inhibition of GCM1 resulted in an increased rate of proliferation, but prevented de novo syncytiotrophoblast formation in syncytially denuded floating villous explants. GCM1 inhibition prevented extravillous differentiation along the invasive pathway in extravillous explants on matrigel. By contrast, forskolin induced expression of GCM1 reduced the rate of proliferation and increased the rate of syncytialization in the floating villous explant model. Our studies show that GCM1 has a distinct role in the maintenance, development and turnover of the human trophoblast. Alterations in GCM1 expression or regulation may explain several aspects of two divergent severe placental insufficiency syndromes, namely preeclampsia and intrauterine growth restriction, which cause extreme preterm birth. PMID- 19219069 TI - PAR bZIP-bik is a novel transcriptional pathway that mediates oxidative stress induced apoptosis in fibroblasts. AB - PAR bZIP (cells knockout for PAR bZIP transcription factors) proteins, thyrotroph embryonic factor (TEF), albumin D-site-binding protein (DBP), and hepatic leukemia factor (HLF), are a family of transcription factors that have been shown to contribute to the expression of genes involved in detoxification and drug metabolism. Recently, we showed that PAR bZIP proteins were able to regulate the BH3-only gene bcl-gS in tumor cells. Here, we have extended the role of these transcription factors in the control of apoptosis executors by analyzing the expression of BH3-only genes in PAR bZIP triple knockout mouse fibroblasts. We found that bik was the only BH3-only gene downregulated in knockout cells. Consistently, transfection of TEF or DBP induces the expression of endogenous bik, regardless of the presence of active p53. Moreover, both promoter-reporter and chromatin immunoprecipitation assays indicate that PAR bZIP proteins activate the bik promoter directly. Treatment with different stress stimuli reveals a higher survival of knockout fibroblasts compared with that of wild-type cells, especially after incubation with H(2)O(2), which suggest that PAR bZIP proteins participate in oxidative stress-induced apoptosis. Furthermore, the apoptotic cell death promoted by treatment with H(2)O(2) can be impaired by reducing the expression of Bik in wild-type fibroblasts or enhanced by the overexpression of Bik in knockout cells. These findings reveal a novel transcriptional pathway relevant in transducing the apoptotic response to oxidative stress. PMID- 19219070 TI - Sox5 can suppress platelet-derived growth factor B-induced glioma development in Ink4a-deficient mice through induction of acute cellular senescence. AB - SOX5 is a member of the high-mobility group superfamily of architectural non histone proteins involved in gene regulation and maintenance of chromatin structure in a wide variety of developmental processes. Sox5 was identified as a brain tumor locus in a retroviral insertional mutagenesis screen of platelet derived growth factor B (PDGFB)-induced mouse gliomas. Here we have investigated the role of Sox5 in PDGFB-induced gliomagenesis in mice. We show that Sox5 can suppress PDGFB-induced glioma development predominantly upon Ink4a-loss. In human glioma cell lines and tissues, we found very low levels of SOX5 compared with normal brain. Overexpression of Sox5 in human glioma cells led to a reduction in clone formation and inhibition of proliferation. Combined expression of Sox5 and PDGFB in primary brain cell cultures caused decreased proliferation and an increased number of senescent cells in the Ink4a-/- cells only. Protein analyses showed a reduction in the amount and activation of Akt and increased levels of p27(Kip1) upon Sox5 expression that was dominant to PDGFB signaling and specific to Ink4a-/- cells. Upon inhibition of p27(Kip1), the effects of Sox5 on proliferation and senescence could be reversed. Our data suggest a novel pathway, where Sox5 may suppress the oncogenic effects of PDGFB signaling during glioma development by regulating p27(Kip1) in a p19(Arf)-dependent manner, leading to acute cellular senescence. PMID- 19219071 TI - Far upstream element-binding protein-1, a novel caspase substrate, acts as a cross-talker between apoptosis and the c-myc oncogene. AB - Far upstream element-binding protein-1 (FBP-1) binds to an upstream element of the c-myc promoter and regulates the c-myc mRNA level. Earlier, FBP-1 was identified as a candidate substrate of caspase-7. Here, we report that FBP-1 is cleaved by executor caspases, both in vitro and during apoptosis. Cleavage occurs at the caspase consensus site (DQPD(74)) located within the classical bipartite nuclear localization signal sequence. In cells subjected to apoptotic stimuli, the caspase-mediated cleavage of FBP-1 leads to its decreased presence in the nucleus, concomitant with the marked downregulation of c-Myc and its various target proteins. By contrast, cells transfected with a non-cleavable mutant of FBP-1 (D74A) maintain higher levels of c-Myc and are protected from apoptosis. On the basis of these results, we suggest that the oncogenic potential of c-Myc is 'switched off' after apoptosis induction as a consequence of the caspase-mediated cleavage of FBP-1. PMID- 19219072 TI - Matrix Metalloproteinase-9 gene induction by a truncated oncogenic NF-kappaB2 protein involves the recruitment of MLL1 and MLL2 H3K4 histone methyltransferase complexes. AB - Constitutive nuclear factor (NF)-kappaB activation in haematological malignancies is caused in several cases by loss of function mutations within the coding sequence of NF-kappaB inhibitory molecules such as IkappaBalpha or p100. Hut-78, a truncated form of p100, constitutively generates p52 and contributes to the development of T-cell lymphomas but the molecular mechanism underlying this oncogenic potential remains unclear. We show here that MMP9 gene expression is induced through the alternative NF-kappaB-activating pathway in fibroblasts and also on Hut-78 or p52 overexpression in fibroblasts as well as in lymphoma cells. p52 is critical for Hut-78-mediated MMP9 gene induction as a Hut-78 mutant as well as other truncated NF-kappaB2 proteins that are not processed into p52 failed to induce the expression of this metalloproteinase. Conversely, MMP9 gene expression is impaired in p52-depleted HUT-78 cells. Interestingly, MLL1 and MLL2 H3K4 methyltransferase complexes are tethered by p52 on the MMP9 but not on the IkappaBalpha promoter, and the H3K4 trimethyltransferase activity recruited on the MMP9 promoter is impaired in p52-depleted HUT-78 cells. Moreover, MLL1 and MLL2 are associated with Hut-78 in a native chromatin-enriched extract. Thus, we identified a molecular mechanism by which the recruitment of a H3K4 histone methyltransferase complex on the promoter of a NF-kappaB-dependent gene induces its expression and potentially the invasive potential of lymphoma cells harbouring constitutive activity of the alternative NF-kappaB-activating pathway. PMID- 19219074 TI - Cooperation between GLI and JUN enhances transcription of JUN and selected GLI target genes. AB - Sustained Hedgehog (HH) signaling is implicated in basal cell carcinoma of the skin and other types of cancer. Here we show that GLI1 and GLI2, the main transcriptional activators of the HH pathway, directly regulate expression of the activator protein 1 (AP-1) family member JUN, a transcription factor controlling keratinocyte proliferation and skin homeostasis. Activation of the JUN promoter by GLI is dependent on a GLI-binding site and the AP-1 sites known to be involved in self-activation of JUN. Transcription of JUN is greatly enhanced in the presence of GLI and requires activated JUN protein. GLI2act is a more potent activator than GLI1 in these experiments and physical interaction with phosphorylated JUN was only detected for GLI2act. The synergistic effect of GLI and JUN extends to the activation of further GLI target genes as shown by shRNA mediated knockdown of JUN in human keratinocytes. Some of these cooperatively activated genes are involved in cell-cycle progression, which is consistent with a significant reduction of the proliferative potential of GLI in the absence of JUN. These results suggest a novel connection between HH/GLI pathway activity and JUN, which may contribute to the oncogenic activity of HH/GLI signaling in skin. PMID- 19219073 TI - Ubiquitination of mammalian AP endonuclease (APE1) regulated by the p53-MDM2 signaling pathway. AB - APE1/Ref-1 is an essential DNA repair/gene regulatory protein in mammals of which intracellular level significantly affects cellular sensitivity to genotoxicants. The apurinic/apyrimidinic endonuclease 1 (APE1) functions are altered by phosphorylation and acetylation. We here report that APE1 is also modified by ubiquitination. APE1 ubiquitination occurred specifically at Lys residues near the N-terminus, and was markedly enhanced by mouse double minute 2 (MDM2), the major intracellular p53 inhibitor. Moreover, DNA-damaging reagents and nutlin-3, an inhibitor of MDM2-p53 interaction, increased APE1 ubiquitination in the presence of p53. Downmodulation of MDM2 increased APE1 level, suggesting that MDM2-mediated ubiquitination can be a signal for APE1 degradation. In addition, unlike the wild-type APE1, ubiquitin-APE1 fusion proteins were predominantly present in the cytoplasm. Therefore, monoubiquitination not only is a prerequisite for degradation, but may also alter the APE1 activities in cells. These results reveal a novel regulation of APE1 through ubiquitination. PMID- 19219075 TI - Efficacy of dasatinib in a chronic myeloid leukemia patient with disease molecular relapse and chronic GVHD after haploidentical BMT: an immunomodulatory effect? PMID- 19219076 TI - Long-term survival after HLA-haploidentical SCT from noninherited maternal antigen-mismatched family donors: impact of chronic GVHD. PMID- 19219077 TI - Limited penetration of posaconazole into cerebrospinal fluid in an allogeneic stem cell recipient with invasive pulmonary aspergillosis. PMID- 19219078 TI - Fatal combined immune hemolytic anemia after double cord blood transplantation in imatinib-resistant CML. PMID- 19219079 TI - NOD2 polymorphisms predict severe acute graft-versus-host and treatment-related mortality in T-cell-depleted haematopoietic stem cell transplantation. AB - Single nucleotide polymorphisms (SNPs) in the NOD2 gene have significant impact on both treatment-related mortality (TRM) and acute GVHD (aGVHD) in haematopoietic stem cell transplantation (HSCT). The effect of these polymorphisms when using T-cell-depleted grafts has been poorly studied. We retrospectively analysed NOD2 polymorphisms in a cohort of 85 patients and donors who received an HLA-identical sibling partially T-cell-depleted HSCT (0.5 x 10(6) CD3+ T cells per kg) following idarubicin-containing conditioning regimens. NOD2 polymorphisms were present in 14 of 85 (16.5%) of patients and 18 of 85 (21%) of donors. The risk of severe aGVHD (grade III-IV) and the 1-year TRM was significantly higher in the presence of NOD2 polymorphisms (hazard ratio (HR) 6.0, P=0.02 for severe aGVHD and HR 3.3, P=0.02 for TRM, respectively) and was most prominent in cases where patient and donor both had a polymorphism (HR 10.5, P=0.002 and HR 3.9, P=0.002). There was also a trend towards increased risk of bacteraemia due to coagulase-negative staphylococci in patients with an NOD2 polymorphism. We conclude that NOD2 polymorphism screening should be used to optimize donor selection and antimicrobial prophylaxis to reduce the occurrence of aGVHD and TRM following allogeneic HSCT. PMID- 19219080 TI - An unusual clinical course of cardiac Ig deposition disease. PMID- 19219081 TI - [Public health and preventive medicine]. PMID- 19219082 TI - [Healthcare research and regional programming in Umbria (Italy)]. AB - This paper discusses the relationship between regional programming and national health research policy, and in particular evaluates the role that intellectual capital plays in innovation processes of knowledge-based organizations. The concepts of intellectual capital and knowledge-based organization are defined in the paper, as these are especially useful when speaking of university, healthcare systems and research and development companies. The paper also examines the various types of healthcare research (basic research, clinical trial, epidemiological research, valutative research) and the role that each type of research plays in healthcare programming at the national and regional levels. PMID- 19219083 TI - [MADEsmart: a web-based system for accessing data and healthcare indicators]. AB - Methods for accessing information have evolved making thus possible the planning of a new generation of web applications. In the Piemonte region, tools for accessing demographic data and main health and epidemiologic indicators are available since several years. A new application named MADEsmart (from the Italian for "Engine for demographic and epidemiological analysis") provides various functions that allow the user to obtain information useful for creating health reports and health profiles at the subregional level. PMID- 19219084 TI - [Evaluation of the quality of hospital medical records in a hospital in Turin (Italy)]. AB - The San Giovanni Battista Hospital in Turin validated a tool for evaluating the quality of hospital patient records. The tool defines the essential contents of patient records, indicators and weights as well as the standard score that must be reached. A pilot study was performed in 2007 to evaluate whether this tool adequately evaluates the quality of hospital patient records in both medical and surgical wards, and whether it can do so in a standardized and repeatable manner. A random sample of 206 medical charts of patients admitted to the San Giovanni Battista Hospital in 2007 was extracted and analysed. The instrument was found to adequately evaluate hospital patient records in a standardisd and repeatable manner. PMID- 19219085 TI - [Health effects of heat waves]. AB - Climate change, characterized by increased mean temperatures and more frequent occurrence of extreme temperatures, may lead to adverse health effects through different and complex mechanisms due to substantial changes in the physical and social environment. The most easily measurable health effects of climate change in high-risk groups are the direct effects of excessively high temperatures. A literature review was performed to identify studies regarding heat waves, risk factors and prevention programmes. A large number of studies performed in Europe during the August 2003 heat wave, and previously in both Europe and North America, showed excess mortality during periods of extreme heat. The role of pollution as a confounding factor remains dubious. Groups at risk include the elderly, the urban population, individuals with impaired health and those with low income. Aging of the population (especially in industrialised nations) and increased urbanization (especially in developing countries) may further increase ill health effects of high temperatures. PMID- 19219086 TI - [Effectiveness of the project "I don't smoke, and you?" against smoking in a secondary school in Italy]. AB - "I don't smoke, and you?" is a project aimed at secondary school students and is part of a wider project called "Free from Smoke", implemented by the Lombardia Region and involving elementary and secondary school students. The project is a controlled non randomized study whose aim is to direct students toward a healthy and smoke-free way of living, by also involving parents and teachers. A total of 11,610 12 year-olds of both sexes were evaluated, 6392 of whom were enrolled in the program and 5218 of whom served as controls and only completed the questionnaire. Three years after the start of the program, a greater increase in the prevalence of smokers was found amongst the group of controls than amongst the enrolled group (108,3% vs +93.8%, p>0.001). The percentage of students who believe that smoking even a small number of cigarettes is harmful, increased in the enrolled group (+0.6%) while it diminished in controls (-2.1%). In addition, a strong association was found between receiving a weekly allowance and smoking habits. In conclusion, positive results were obtained as regards the prevalence of smokers and the perceived risk of smoking; however parents' attitude towards smoking and the availability of a weekly allowance were found to have a strong influence in students' smoking habits. PMID- 19219087 TI - Factors influencing student nurses' perception of their clinical guides: a questionnaire survey. AB - Supporting student nurses whilst undertaking clinical training is an important activity for both educators and students, yet there is little consensus in the literature as to what constitutes support. Also, it is difficult to find a robust instrument that provides a valid measure of the quality of the clinical learning environment in different contexts. The aim of this study was to identify factors that influence student nurses' perception of their clinical tutors. A two-part questionnaire was administered to students of a Nursing School in a large city in northern Italy; 143 students participated in the study. In replying to the questions, student nurses were asked to refer to their last one-month training period of the year 2004. Analysis of study results showed that students' perceived satisfaction with their clinical guide was greater in cases where tutors used the critical thinking method. A one-to-one relation between tutor and student was not found to be essential in assuring a satisfactory clinical experience for nursing students. PMID- 19219088 TI - [Paternalism and preventive medicine]. PMID- 19219090 TI - [Acute treatment of specialist education]. PMID- 19219089 TI - [Invasive pneumococcal diseases in children and surveillance systems in Italy]. AB - Streptococcus pneumoniae is the causative agent of invasive diseases and of localized infections in children and older people. World and European epidemiological data show differences in disease incidences and in prevalences of pathogenic serotypes. Epidemiology changed after the introduction of conjugate vaccines for infants even if the still inadequate surveillance systems do not allow a comparison of the data. In this paper the Italian available epidemiological data and surveillance systems are rewied considering problems related to the emergence of new or drug-resistant strains and the advantages awaited from the new vaccines. PMID- 19219091 TI - [More than centimeters and grams]. PMID- 19219092 TI - [Growth references for Norwegian children]. AB - BACKGROUND: The growth charts currently used in Norway, are based on measurements from the 1970s and 80s. New data are available from the Bergen Growth Study collected in 2003 - 6. In 2006, WHO published international charts for 0-5 year old children. MATERIAL AND METHODS: New growth charts based on data from the Bergen Growth Study and the Medical Birth Registry of Norway are presented for children aged 0-19 years. These were compared with existing references and with the WHO curves. RESULTS: Norwegian children aged 0-4 years have length, height and weight measurements that are only marginally different from those in the Norwegian growth charts in current use. In older children there has been an increase in the 50-percentile for height up to 3.4 cm in boys and 2.5 cm in girls. For children older than four years, weight for height has increased, especially for the upper percentiles. The percentile lines in the new Norwegian reference are generally positioned above the WHO standard for weight at birth, and for length/height, weight and head circumference in the age group 6 months to 5 years. INTERPRETATION: The secular trends in growth mirror the need for new charts. The fact that Norwegian children differ from the WHO standards may reflect population differences relating to environment or growth potential between the populations. PMID- 19219093 TI - [Substitution treatment of drug addicts during pregnancy: consequences for the children?]. AB - BACKGROUND: Substitution treatment of opioid-dependent addicts was introduced in Norway in 1998. During the last 10 years, approximately 150 infants have been born to mothers taking part in this programme. MATERIAL AND METHODS: 10 mothers, who took part in the substitution treatment programme, gave birth to 15 infants at Haukeland University Hospital in the period 1999-2005. The infants were observed and monitored at the Department of Pediatrics, Haukeland University Hospital. RESULTS: During pregnancy, six of the infants were only exposed to opiates, i.e methadone or buprenorphine. Eight infants were also exposed to heroine, benzodiazepines or cannabis. As a group, these infants had lower birth weight than the national average. 14 of the 15 children developed neonatal abstinence syndrome (NAS), 10 needed treatment and two children died from sudden infant death syndrome (SIDS). Long-term follow-up showed that six of 13 children had normal psychomotor development, five had various degrees of delayed psychomotor development and two children had symptoms indicating a hyperkinetic disorder. Five children were in foster care. INTERPRETATION: Infants of women included in substitution treatment programmes for drug addicts are at high risk compared to infants of women without such addiction. For the newborn, NAS was a frequent complication. The study also showed that symptoms of hyperkinetic disorder and delayed psychomotor development were common. Children who had been exposed to opiates in combination with additional drugs seemed to have a high risk of delayed development and behaviour disorders. As they get older many were placed in foster care, despite well-coordinated, multidisciplinary treatment for the mother. PMID- 19219094 TI - [Catheter ablation of tachyarrhythmias in children and adolescents]. AB - BACKGROUND: Catheter ablation has been increasingly applied in children and adolescents with tachyarrhythmias. The aim of this article is to assess the results of ablation therapy of tachycardias in patients below 18 years of age at Haukeland University Hospital. MATERIAL AND METHODS: 141 patients (70 boys and 71 girls, aged 5-17 (13.5 +/- 3.5 ) years with tachyarrhythmias underwent an electrophysiologic study and catheter ablation in the period 1992-2007. RESULTS: Ablation was successfully performed in 138/141 (98%) patients., The procedure was repeated (3 patients twice) until the arrhythmia substrate disappeared in 16 of 138 patients. 81/141 (57%) patients had accessory pathways; 52 (37%) had double atrioventricular nodal pathways, 48 had concealed and 33 patients had overt (classical Wolff-Parkinson-White-syndrome) atrioventricular pathways. 8 (6%) patients had other atrial or ventricular tachyarrhythmias and 4 (3%) had organic heart disease. Use of a 3D mapping system was decisive for success for ablation in patients with complex cardiac diseases. Procedure-related complications were observed in 2/141 (1.4%) patients of whom one had a temporary third degree and one had a permanent first-degree atrioventricular block which did not entail further treatment. CONCLUSION: Catheter ablation of tachycardia in children and adolescents is a safe treatment method with a high success rate and few complications and should be preferred before drug therapy. PMID- 19219095 TI - [Cognitive failure in terminal kidney disease]. AB - BACKGROUND: Renal failure occurs more frequently among the elderly and elderly patients comprise an increasing proportion of dialysis patients. The objective of this paper is to review the frequency of cognitive impairment among elderly people in dialysis, and discuss causes and consequences of cognitive impairment and dementia in patients with end-stage renal disease. Prevention and treatment of cognitive impairment are discussed. MATERIAL AND METHODS: The review is based on literature retrieved from a search of Medline and other web-sites, review papers and clinical experience. RESULTS AND INTERPRETATION: Cognitive impairment occurs in 20-87% of patients in haemodialysis, and the frequency is significantly more increased than that for control groups. The etiology is multifactorial, including the primary renal disease, comorbidity, the effect of uraemia and treatment-related factors. Potentially reversible factors such as anaemia and treatment complications may contribute to cognitive impairment in these patients. Cognitive impairment has major clinical consequences for compliance, resource use and the prognosis for patients with renal failure, and is often not detected by the clinicians. The authors propose that cognitive assessment should be included in the routine evaluation of elderly patients with renal failure, with potential implications for the treatment and quality of treatment for these patients. PMID- 19219096 TI - [Treatment practice in primary hyperparathyroidism]. AB - BACKGROUND: A recent study found considerable regional differences in treatment of primary hyperparathyroidism in Norway. There is no consensus on specific indications for operation in these patients. We surveyed opinions among Norwegian endocrine surgeons and endocrinologists on the indications for surgical treatment of primary hyperparathyroidism. MATERIAL AND METHODS: A questionnaire on preoperative evaluation, indications for surgery and treatment of patients with primary hyperparathyroidism was sent to the chief consultants of surgical departments that operated on parathyroid glands in 2005. The questionnaire was also sent to endocrinologists at the same hospitals. RESULTS: In 2006, 415 parathyroid gland operations were performed in 17 Norwegian hospitals, with a median of 18 operations per hospital. A total of 46 surgeons operated on the parathyroid glands, with a median of two surgeons per hospital. Hospitals differed with respect to preoperative evaluations and indications for operative treatment; but these differences did not coincide with regional differences in the frequency of parathyroid surgery. There was a good correlation between endocrine surgeons and endocrinologists on the indications for surgery in primary hyperparathyroidism, but neither group adhered unconditionally to the international guidelines for surgical treatment of patients with primary hyperparathyroidism. Patients in the hospitals that operated most frequently were initially diagnosed in a surgical department. INTERPRETATION: Our survey did not reveal differences that could explain the large regional variations in the frequency of parathyroid surgery. PMID- 19219098 TI - [When the patient refuses treatment]. PMID- 19219097 TI - [Cranberries for prevention of recurrent urinary tract infections]. AB - BACKGROUND: Cranberries have been used for prevention and treatment of urinary tract infections for decades. The berries contain proanthocyanidins that may reduce the susceptibility to infection by preventing bacteria from attaching to uroepithelial cells. Several clinical trials have been published during recent years. This article reviews documentation of cranberries on clinical effect, adverse events, drug interactions and use during pregnancy and lactation. MATERIAL AND METHODS: Clinical effects of cranberries have been assessed based on the Cochrane review from January 2007 and literature on clinical trials retrieved from a systematic search of PubMed and Embase (from 1 January 2007 to 29 October 2008) with the search terms "cranberry", "Vaccinium macrocarpon", "Vaccinium oxycoccus". RESULTS AND INTERPRETATION: Some evidence exists on cranberries' preventive effect on recurrent symptomatic urinary tract infections in women. The evidence is inconclusive for children, men and older people (both men and women). Studies of people with neuropathic bladder are contradictory. Most of the clinical trials published have several flaws and have not used standardised products. More evidence is needed to determine the optimum dosage, method of administration and the minimum length of treatment. Cranberries should not be used during pregnancy and lactation due to lack of safety data. Further, properly designed studies with standardised products and relevant outcomes are needed. PMID- 19219100 TI - [Misleading about fluid therapy in practice guidelines]. PMID- 19219101 TI - [Do-not-resuscitate orders in a stroke unit]. PMID- 19219102 TI - [Societal insult to elderly physicians]. PMID- 19219103 TI - [Wrong about euthanasia in Netherlands]. PMID- 19219104 TI - [Tobacco sale in hospitals--sad and strange chapter in Norwegian medicine]. PMID- 19219105 TI - [Neonatal pulse oximetry]. PMID- 19219108 TI - [Epilepsy campaign in China shows positive results]. PMID- 19219110 TI - 10-watt, pure continuous-wave, polycrystalline Cr2+:ZnS laser. AB - We demonstrate, for the first time, 10 W, Er-fiber laser pumped, pure CW, thermally diffusion doped, polycrystalline Cr(2+):ZnS laser operating at 2380 nm. We also show Littrow-grating, "single-knob", wavelength tuning of the laser spanning 1940-2780 nm spectral range with the maximum output power of 7.4 W near the central wavelength of 2400 nm and above 2 W over 1970-2760 nm wavelength range. The laser performs with 40% real optical- and 43% slope efficiency, and shows no output power roll-off up to the highest available incident pump power of 27 W. PMID- 19219111 TI - A vector radiative transfer model for coupled atmosphere and ocean systems based on successive order of scattering method. AB - A vector radiative transfer model has been developed for coupled atmosphere and ocean systems based on the Successive Order of Scattering (SOS) Method. The emphasis of this study is to make the model easy-to-use and computationally efficient. This model provides the full Stokes vector at arbitrary locations which can be conveniently specified by users. The model is capable of tracking and labeling different sources of the photons that are measured, e.g. water leaving radiances and reflected sky lights. This model also has the capability to separate florescence from multi-scattered sunlight. The delta - fit technique has been adopted to reduce computational time associated with the strongly forward peaked scattering phase matrices. The exponential - linear approximation has been used to reduce the number of discretized vertical layers while maintaining the accuracy. This model is developed to serve the remote sensing community in harvesting physical parameters from multi-platform, multi-sensor measurements that target different components of the atmosphere-oceanic system. PMID- 19219112 TI - Optimization of 3-hole-assisted PMMA optical fiber with double cladding for UV induced FBG fabrication. AB - We propose a new hole-assisted polymer optical fiber design to eliminate the influence of dopant diffusion and to increase the UV writing efficiency in fiber Bragg grating inscription. The optical waveguide is formed inside a solid core surrounded by a ring of 3 large air holes in enhanced UV photosensitive PMMA with double-cladding. We determined a map of the single-mode and multi-mode phase transitions using a finite-element- based vectorial optical mode solver. We obtained a wide range of geometrical configuration for the single-transverse-mode (HE11) propagation in the visible. The design is optimized to operate at the minimum optical loss wavelengths of 580 nm and 770 nm. PMID- 19219113 TI - Three-dimensional subwavelength confinement of light with dielectric microspheres. AB - Dielectric microspheres are shown to be capable of confining light in a three dimensional region of subwavelength dimensions when they are illuminated by tightly focused Gaussian beams. We show that a simple configuration, not involving resonances, permits one to reach an effective volume as small as 0.6 (lambda/n)(3). It is shown that this three-dimensional confinement arises from interferences between the field scattered by the sphere and the incident Gaussian beam containing high angular components. PMID- 19219114 TI - Solid-state single photon sources: the nanowire antenna. AB - We design several single-photon-sources based on the emission of a quantum dot embedded in a semiconductor (GaAs) nanowire. Through various taper designs, we engineer the nanowire ends to realize efficient metallic-dielectric mirrors and to reduce the divergence of the far-field radiation diagram. Using fully vectorial calculations and a comprehensive Fabry-Perot model, we show that various realistic nanowire geometries may act as nanoantennas (volume of approximately 0.05 lambda(3)) that assist funnelling the emitted photons into a single monomode channel. Typically, very high extraction efficiencies above 90% are predicted for a collection optics with a numerical aperture NA=0.85. In addition, since no frequency-selective effect is used in our design, this large efficiency is achieved over a remarkably broad spectral range, Deltalambda=70 nm at lambda=950 nm. PMID- 19219115 TI - Wyckoff positions and the expression of polarization singularities in photonic crystals. AB - We reveal the fundamental relation between linear photonic crystal symmetries and the local polarization states of its Bloch modes, in particular the location and nature of polarization singularities as established by rigorous group theoretic analysis, encompassing the full system symmetry. This is illustrated with the fundamental transverse electric mode of a two-dimensional hexagonal photonic crystal, in the vanishing contrast limit and at the K point. For general Wyckoff positions within the fundamental domain, the transformation of a local polarization state is determined by the nature of the symmetry operations that map to members of its crystallographic orbit. In particular, the site symmetries that correspond to specific Wyckoff positions constrain the local polarization state to singular character--circular, linear or disclination. Moreover, through the application of a local symmetry transformation relation, and the group's character table, the precise natures of the singularities may be determined from self-consistency arguments. PMID- 19219116 TI - Hybrid resonant phenomena in a SRR/YIG metamaterial structure. AB - We consider the hybridization of the resonance of a SRR metamaterial with the gyromagnetic material resonance of yittrium iron garnet (YIG) inclusions. The combination of an artificial structural resonance and natural material resonance generates a unique hybrid resonance that can be harnessed to make tunable metamaterials and further extend the range of achievable electromagnetic materials. A predictive analytic model is applied that accurately describes the characteristics of this SRR/YIG hybridization. We suggest that this hybridization has been observed in experimental data presented by Kang et al. [Opt. Express, 16, 8825 (2008)] and present numerical simulations to support this assertion. In addition, we investigate a design for optimizing the SRR/YIG structure that shows strong hybridization with a minimum amount of YIG material. PMID- 19219117 TI - Effects of bulk particle characteristics on backscattering and optical closure. AB - Optical closure is essential for the determination of biogeochemical properties from ocean color remote sensing information. Mie scattering theory, a radiative transfer model, and a semi-analytical inversion algorithm were used to investigate the influence of particles and their properties on optical closure. Closure results were generally poor. Absorption coefficient (a(t)) inversions were more accurate for moderate particle size distribution slopes (3.50 < or = xi < or = 3.75). The degree of success in the derivation of the backscattering coefficient (b(bp)) was highest at moderate indices of refraction (1.15 < or = n(p) < or = 1.20) and high values of xi(> 3.75). Marked improvements in the estimates of b(bp) were enabled by a priori knowledge of bbp at one wavelength. At moderate values of n(p), derivations of a(t) and b(bp) were within 25% of Mie modeled values when Gershun's relationship was used in combination with the semi analytical algorithm. PMID- 19219118 TI - Shrinkage of microstructures produced by two-photon polymerization of Zr-based hybrid photosensitive materials. AB - An investigation of the shrinking behaviour of a zirconium-based sol-gel composite micro-structured by two-photon polymerization is presented and a simple, straightforward methodology allowing the evaluation of shrinkage is suggested. It is shown that volume reduction is directly related to the average laser power (irradiation dose) used for the microfabrication and becomes a critical issue near the polymerization threshold. It is demonstrated that this shrinkage can be employed beneficially to improve the structural resolution. This is demonstrated by the presence of stopbands in the photonic crystal nanostructures fabricated with controlled volume reduction. Well above the polymerization threshold, the studied material exhibits remarkably low shrinkage. Therefore, no additional effort for the pre-compensation of distortion and for the improvement of structural stability is required. PMID- 19219119 TI - Optical design of the proposed Australian International Gravitational Observatory. AB - Marginally stable power recycling cavities are being used by nearly all interferometric gravitational wave detectors.With stability factors very close to unity the frequency separation of the higher order optical modes is smaller than the cavity bandwidth. As a consequence these higher order modes will resonate inside the cavity distorting the spatial mode of the interferometer control sidebands. Without losing generality we study and compare two designs of stable power recycling cavities for the proposed 5 kilometer long Australian International Gravitational Observatory (AIGO), a high power advanced interferometric gravitational wave detector. The length of various optical cavities that form the interferometer and the modulation frequencies that generate the control sidebands are also selected. PMID- 19219120 TI - Performance evaluation of optical packet switches equipped with heterogeneous wavelength converters. AB - An optical packet switch that shares both limited range and full range wavelength converters for contention resolution is proposed with the aim to guarantee a high conversion cost saving. To optimally dimension the number and the conversion range of the wavelength converters, an analytical model, validated by simulation, is introduced. Numerical results show that the proposed switch architecture allows for a conversion cost saving in the order of 90% with respect to a traditional architecture in which only shared full range wavelength converters are used. PMID- 19219121 TI - Breakthrough switching speed with an all-optical chalcogenide glass chip: 640 Gbit/s demultiplexing. AB - We report the first demonstration of error-free 640 Gbit/s demultiplexing using the Kerr non-linearity of an only 5 cm long chalcogenide glass waveguide chip. Our approach exploits four-wave mixing by the instantaneous nonlinear response of chalcogenide. Excellent performance is achieved with only 2 dB average power penalty and no indication of error-floor. Characterisation of the FWM efficiency for the chalcogenide waveguide is given and confirms the good performance of the device. PMID- 19219122 TI - Greatly enhanced slow and fast light in chirped pulse semiconductor optical amplifiers: theory and experiments. AB - Chirped pulse scheme is shown to be highly effective to attain large tunable time shifts via slow and fast light for an ultra-short pulse through a semiconductor optical amplifier (SOA). We show for the first time that advance can be turned into delay by simply reversing the sign of the chirp. A large continuously tunable advance-bandwidth product (ABP) of 4.7 and delay-bandwidth product (DBP) of 4.0 are achieved for a negatively and positively chirped pulse in the same device, respectively. We show that the tunable time shift is a direct result of self-phase modulation (SPM). Theoretical simulation agrees well with experimental results. Further, our simulation results show that by proper optimization of the SOA and chirper design, a large continuously tunable DBP of 55 can be achieved. PMID- 19219123 TI - SPP-associated dual left-handed bands and field enhancement in metal-dielectric metal metamaterial perforated by asymmetric cross hole arrays. AB - Dual-band left-handed transmissions in the near infrared frequencies through the metal-dielectric-metal metamaterial perforated with an array of asymmetric cross holes are demonstrated. It is shown that the left-handed bands originate from the SPP-associated magnetic response excited by different polarized light and their frequencies can be tuned by the arm's length or width of the cross-gaps. The structures are further optimized at 1.064 microm laser light excitation for elucidating the mechanism and possible application in surface enhanced Raman spectroscopy in sandwiched architectures. This study provides valuable information for the design of compact optical devices with dual left-handed bands in a single structure and may also pave the way toward stable and reproducible substrate design for surface enhanced Raman spectroscopy. PMID- 19219124 TI - Comparison of dispersive mirrors based on the time-domain and conventional approaches, for sub-5-fs pulses. AB - Dispersive mirrors based on time-domain approach are compared with mirrors resulting from conventional phase target designs. Phase targets have been applied to complementary-pair dispersive mirrors, used for sub-5-fs pulse compression. While the phase approach has hither to afforded the best performance for the shortest pulses, our new approach, based on time-domain targets and tailored for a specific input spectrum, appears to provide comparable performance for pulse compression for a pulse duration 4.6 fs. Experimental studies using dispersive mirrors made to both designs are described. PMID- 19219125 TI - Compact architecture for power scaling bounce geometry lasers. AB - We demonstrate the compact high-power scaling of bounce geometry lasers with a new dual-pumped folded amplifier design. A Q-switched laser oscillator built with this amplifier is shown to produce over 30 W of average power from 80 W of pump power at up to 600 kHz repetition rate. In a master-oscillator power-amplifier (MOPA) configuration using the dual-pumped amplifier, we demonstrate over 100 W of output power from 250 W of pump power. We also demonstrate very high repetition rate Q-switching (1.7 MHz) of the master oscillator. PMID- 19219126 TI - Electromagnetic force and torque on magnetic and negative-index scatterers. AB - We derive the analytic expressions of the electromagnetic force and torque on a dipolar particle, with arbitrary dielectric permittivity and magnetic permeability. We then develop a general framework, based on the coupled dipole method, for computing the electromagnetic force and torque experienced by an object with arbitrary shape, dielectric permittivity and magnetic permeability. PMID- 19219127 TI - Laser expander design of highly efficient Blu-ray disc pickup head. AB - We present a collimator design with a horizontal beam expander for a Blu-ray Disc pickup head. The design transforms the shape of a laser diode beam from an elliptical into circular which achieves higher efficiency in the pickup head system. In this research, we suggest two ways to do the beam expander. One way is to use two cylindrical lenses, and the other is to use two prisms. PMID- 19219128 TI - Design and demonstration of compact, wide bandwidth coupled-resonator filters on a siliconon- insulator platform. AB - We design and fabricate a compact third-order coupledresonator filter on the silicon-on-insulator platform with focused application for on-chip optical interconnects. The filter shows a large flat bandwidth (3dB 3.3nm), large FSR (approximately 18nm), more than 18dB out-of-band rejection at the drop port and more than 12 dB extinction at the through port, as well as a negligible drop loss (<0.5dB) within a footprint of 0.0004 mm(2). PMID- 19219129 TI - Designing efficient zero calibration point for phase-sensitive surface plasmon resonance biosensing. AB - This work is related to the development of phase-sensitive methodologies in Surface Plasmon Resonance (SPR) biosensing. We take advantage of a specific angular dependence of phase of light, reflected under SPR geometry, on parameters of the SPR-supporting metal, and propose a polarimetry-based methodology to easily determine the optimal calibration zero point, corresponding to the maximal phase sensitivity. The proposed methodology can significantly facilitate the calibration of the system in field and multi-channel sensing, broaden the dynamic range, as well as contribute to the development of feedback loops. PMID- 19219130 TI - Properties and stability limits of an optimized mode-locked Yb-doped femtosecond fiber laser. AB - Stable mode-locked operation in a simple normal-cavity-dispersion laser oscillator that consists of only Yb-doped fiber and saturable absorber is studied. The Yb-doped fiber design parameters: group velocity dispersion (GVD), nonlinearity coefficient, bandwidth (Yb-BW), length and gain are considered to be the controlling parameters of the laser cavity. The pulse characteristics such as the temporal width, spectrum and pulse energy as a function of these elements are reported here. A pulse spectrum transition from M-like to Pi-like and then to parabolic-like shape is observed with different values of the controlling parameters which are similar to that has been observed before in a solid-state laser. The stability limits in the domain of the Yb-BW and length are studied. The stability dependence on GVD, nonlinearity coefficient and gain of the Yb doped fiber are elucidated. Moreover, the pulse instability dyn ics beyond stability limits are found to be similar to that reported before for similariton. PMID- 19219131 TI - 3-dimensional local field polarization vector mapping of a focused radially polarized beam using gold nanoparticle functionalized tips. AB - We have measured local electric field polarization vectors in 3-dimensional space on the nanoscale. A radial polarized light is generated by using a radial polarization converter and focused by an objective lens. Gold nanoparticle functionalized tips are used to scatter the focused field into the far-field region. Two different methods, rotational analyzer ellipsometry and Stokes parameters, are used in determining the polarization state of the scattered light. Two methods give consistent results with each other. Three dimensional local polarization vectors could be reconstructed by applying back transformation of the fully characterized polarizability tensor of the tip. PMID- 19219132 TI - Separate spatial Holographic-Hamiltonian soliton pairs and solitons interaction in an unbiased series photorefractive crystal circuit. AB - This paper presents calculations for an idea in photorefractive spatial soliton, namely, a dissipative holographic soliton and a Hamiltonian soliton in one dimension form in an unbiased series photorefractive crystal circuit consisting of two photorefractive crystals of which at least one must be photovoltaic. The two solitons are known collectively as a separate Holographic-Hamiltonian spatial soliton pair and there are two types: dark-dark and bright-dark if only one crystal of the circuit is photovoltaic. The numerical results show that the Hamiltonian soliton in a soliton pair can affect the holographic one by the light induced current whereas the effect of the holographic soliton on the Hamiltonian soliton is too weak to be ignored, i.e., the holographic soliton cannot affect the Hamiltonian one. PMID- 19219133 TI - A full vectorial model for pulse propagation in emerging waveguides with subwavelength structures part I: Kerr nonlinearity. AB - The propagation of pulses through waveguides with sub-wavelength features, inhomogeneous transverse structure, and high index contrast cannot be described accurately using existing models in the presence of nonlinear effects. Here we report the development of a generalised full vectorial model of nonlinear pulse propagation and demonstrate that, unlike the standard pulse propagation formulation, the z-component of guided modes plays a key role for these new structures, and results in generalised definitions of the nonlinear coefficient gamma, Aeff , and mode orthognality. While new definitions reduce to standard definitions in some limits, significant differences are predicted, including a factor of approximately 2 higher value for gamma, for emerging waveguides and microstructured fibers. PMID- 19219134 TI - A two-state Raman coupler for coherent atom optics. AB - We present results on a Raman laser-system that resonantly drives a closed two photon transition between two levels in different hyperfine ground states of (87)Rb. The coupler is based on a novel optical design for producing two phase coherent optical beams to drive a Raman transition. Operated as an outcoupler, it produces an atom laser in a single internal atomic state, with the lower divergence and increased brightness typical of a Raman outcoupler. Due to the optical nature of the outcoupling, the two-state outcoupler is an ideal candidate for transferring photon correlations onto atom-laser beams. As our laser system couples just two hyperfine ground states, it has also been used as an internal state beamsplitter, taking the next major step towards free space Ramsey interferometry with an atom laser. PMID- 19219135 TI - Ultrafast carrier kinetics in exfoliated graphene and thin graphite films. AB - Time-resolved transmissivity and reflectivity of exfoliated graphene and thin graphite films on a 295 K SiO(2)/Si substrate are measured at 1300 nm following excitation by 150 fs, 800 nm pump pulses. From the extracted transient optical conductivity we identify a fast recovery time constant which increases from approximately 200 to 300 fs and a longer one which increases from 2.5 to 5 ps as the number of atomic layers increases from 1 to approximately 260. We attribute the temporal recovery to carrier cooling and recombination with the layer dependence related to substrate coupling. Results are compared with related measurements for epitaxial, multilayer graphene. PMID- 19219136 TI - Rayleigh anomaly-surface plasmon polariton resonances in palladium and gold subwavelength hole arrays. AB - Surface plasmon polaritons (SPPs) and Rayleigh anomalies (RAs) are two characteristic phenomena exhibited by periodic grating structures made of plasmonic materials. For Au subwavelength hole arrays, SPPs and RAs from opposite sides of the film can interact under certain conditions to produce highly intense, narrow spectral features called RA-SPP resonances. This paper reports how RA-SPP effects can be achieved in subwavelength hole arrays of Pd, a weak plasmonic material. Well-defined resonances are observed in measured and simulated optical transmission spectra with RASPP peaks as narrow as 45 nm (FWHM). Dispersion diagrams compiled from angle-resolved spectra show that RA-SPP resonances in Pd hole arrays shift in wavelength but do not decrease significantly in amplitude as the excitation angle is increased, in contrast with RA-SPP peaks in Au hole arrays. The apparent generality of the RA-SPP effect enables a novel route to optimize resonances in non-traditional plasmonic media. PMID- 19219137 TI - Tunable Yb:KYW laser using a transversely chirped volume Bragg grating. AB - We demonstrate a diode-pumped Yb:KYW laser, where the wavelength is controlled by a transversely chirped volume-Bragg-grating, also used as the laser output coupler. By translating the grating, the wavelength could be tuned from 997 nm to 1016 nm, with a spectral bandwidth of 13 GHz and a power up to 3.3 W. The laser cavity is simple and compact, and the tuning method is generally applicable to many other laser systems. PMID- 19219138 TI - Axial strain dependence of all-fiber acousto-optic tunable filters. AB - We report the axial strain dependence of two types of all-fiber acousto-optic tunable filters based on flexural and torsional acoustic waves. Experimental observation of the resonant wavelength shift under applied axial strain could be explained by theoretical consideration of the combination of acoustic and optical effects. We discuss the possibility of suppressing the strain effect in the filters, or conversely, the possibility of using the strain dependence for wavelength tuning or strain sensors. PMID- 19219140 TI - Channel and wedge plasmon modes of metallic V-grooves with finite metal thickness. AB - We investigate numerically the effect of a finite metal film thickness on the propagation characteristics of the channel Plasmon polariton (CPP) and wedge plasmon polariton (WPP) modes, both in a symmetric and asymmetric environment. We observe that decreasing the metal thickness results in an improvement of the field localization near the groove tip and an increase of the losses for both types of mode. This behavior stems from the typical symmetric charge distribution of both modes across the metal film. When considering an asymmetric dielectric environment, the CPP mode is found to evolve into short range Plasmon modes propagating along the groove walls, in contrast to the WPP mode which remains essentially confined at the tip apex. These results can be useful to tailor the properties of such plasmon modes, using the metal thickness as the variable parameter. PMID- 19219139 TI - Carbon nanotube films for ultrafast broadband technology. AB - Mode-locked sub-picosecond operation of Yb-, Er- and Tm:Hodoped fiber lasers operating at 1.05 microm, 1.56 microm and 1.99 microm, respectively, is demonstrated using the same sample carbon nanotube-based saturable absorber mirror. A mesh of single-walled carbon nanotubes was deposited on an Ag-mirror using a one-step dry-transfer contact press method to combine broadband saturable absorption and high reflectance properties. The novel fabrication method of the polymer-free absorber and device parameters determined using nonlinear reflectivity measurement are described in detail. To our knowledge the observed operation bandwidth of approximately 1 microm is the broadest reported to date for a single carbon nanotube-based saturable absorber. PMID- 19219141 TI - In-fiber common-path optical coherence tomography using a conical-tip fiber. AB - Common-path optical coherence tomography (CPOCT) is known to reduce group velocity dispersion and polarization mismatch between the reference and the sample arm as both arms share the same physical path. Existing implementations of CPOCT typically require one to incorporate an additional cover glass within the beam path of the sample arm to provide a reference signal. In this paper, we aim to further reduce this step by directly making use of the back-reflected signal, arising from a conical lens-tip fiber, as a reference signal. The conical lens, which is directly manufactured onto the optical fiber tip via a simple selective chemical etching process, fulfils two functions acting as both the imaging lens and the self-aligning reference plane. We use a Fourier-domain OCT system to demonstrate the feasibility of this technique upon biological tissue. An in-fiber CPOCT technique may prove potentially useful in endoscopic OCT imaging. PMID- 19219142 TI - Fabrication and characterization of Teflon-bonded periodic GaAs structures for THz generation. AB - A novel technique for low-temperature bonding of GaAs wafers using an interboundary Teflon film is developed. A fabricated stack of ten 25x25 mm2 diced wafers demonstrated 75% transmission of a 10 microm CO2 laser beam. Modeling of these Teflon-bonded (TB) structures as sequences of Fabry-Perot etalons gives a good agreement with transmission measurements. A 20x20 mm2 quasi-phase matched structure of five wafers pumped by CO2 laser lines generated the narrow-band THz radiation at a wavelength of 343 microm via a difference frequency mixing process. PMID- 19219143 TI - Ultra-low-loss optical fiber enabling purely passive 10 Gb/s PON systems with 100 km length. AB - We demonstrate time division multiplexing (TDM) and wavelength division multiplexing/TDM (WDM/TDM) long reach 10 Gb/s passive optical network (PON) architectures of 100 km reach with no infield amplification or dispersion compensation. The purely passive nature of the 100 km systems is enabled by the use of ultra-low-loss optical fiber with average attenuation of 0.17 dB/km and downstream transmission with a 10 Gb/s signal modulated with the duobinary format. The high tolerance of duobinary to dispersion, stimulated Brillouin scattering (SBS), and self-phase modulation (SPM) are all key factors to achieving good system performance at this distance, as is the significantly reduced loss from the ultra-low-loss fiber. We show that this combination of fiber and downstream signal format allow split ratios up to 1:128 for both system architectures. The achievable split ratio is reduced for standard single-mode fiber and/or use of an NRZ modulated downstream signal. Standard strength forward error correction (FEC) is used for the WDM/TDM system but is not required for the TDM system. PMID- 19219144 TI - Computational signal-to-noise ratio analysis for optical quadrature microscopy. AB - Optical quadrature microscopy (OQM) was invented in 1997 to reconstruct a full field image of quantitative phase, and has been used to count the number of cells in live mouse embryos. Here we present a thorough SNR analysis that incorporates noise terms for fluctuations in the laser, aberrations within the individual paths of the Mach-Zehnder interferometer, and imperfections within the beamsplitters and CCD cameras to create a model for the resultant phase measurements. The current RMS error of the OQM phase images has been calculated to be 0.08 radians from substituting images from the instrumentation into the model. PMID- 19219145 TI - Numerical study of effects of light polarization, scatterer sizes and orientations on near-field coherent anti-Stokes Raman scattering microscopy. AB - We employ the finite-difference time-domain (FDTD) technique as a numerical approach to studying the effects of polarization, scatterers' sizes and orientations on near-field coherent anti-Stokes Raman scattering (CARS) microscopy imaging. The results show that to acquire better image contrast and larger near-field CARS signals, the scatterers with diameters of less than three eighths of the pump field wavelength (lambda(p)) are preferable to be oriented along the polarization direction of the excitation light fields. It is also found that when the scatterers' sizes are smaller than half a wavelength of the pump field, the perpendicular polarization component of the induced near-field CARS radiations is strictly confined within the regions at the scatterer-water interface or the subsurface of scatterers, yielding a high image contrast (up to 200) with a spatial resolution of lambda(p)/16. This study indicates that perpendicular polarization components of near-field CARS microscopy could be used to uncover very fine structures of intra- and/or inter- cellular organelles in cells with nanoscale resolutions. PMID- 19219146 TI - Stable mode sorting by two-dimensional parity of photonic transverse spatial states. AB - We describe a mode sorter for two-dimensional parity of transverse spatial states of light based on an out-of-plane Sagnac interferometer. Both Hermite-Gauss (HG) and Laguerre-Gauss (LG) modes can be guided into one of two output ports according to the two-dimensional parity of the mode in question. Our interferometer sorts HG(nm) input modes depending upon whether they have even or odd order n+m; it equivalently sorts LG(l)(p) modes depending upon whether they have an even or odd value of their orbital angular momentum l. It functions efficiently at the single-photon level, and therefore can be used to sort single photon states. Due to the inherent phase stability of this type of interferometer as compared to those of the Mach-Zehnder type, it provides a promising tool for the manipulation and filtering of higher order transverse spatial modes for the purposes of quantum information processing. For example, several similar Sagnacs cascaded together may allow, for the first time, a stable measurement of the orbital angular momentum of a true single-photon state. Furthermore, as an alternative to well-known holographic techniques, one can use the Sagnac in conjunction with a multi-mode fiber as a spatial mode filter, which can be used to produce spatial-mode entangled Bell states and heralded single photons in arbitrary first-order (n+m = 1) spatial states, covering the entire Poincar e sphere of first-order transverse modes. PMID- 19219148 TI - Ghost imaging with twisted Gaussian Schell-model beam. AB - Based on the classical optical coherence theory, ghost imaging with twisted Gaussian Schell-model (GSM) beams is analyzed. It is found that the twist phase of the GSM beam has strong influence on ghost imaging. As the absolute value of the twist factor increases, the ghost image disappears gradually, but its visibility increases. This phenomenon is caused by the fact that the twist phase enhances the transverse spatial coherence of the twisted GSM beam on propagation. PMID- 19219149 TI - Backward propagating slow light in inverted plasmonic taper. AB - The negative dispersion of the TM1 mode of a thin plasmonic gap, occurring at frequencies exceeding the surface plasmon frequency, is assigned by causality to be a backward wave. This negative index mode, is also slow-light--having small positive group velocity and is exhibiting inverse geometrical cutoff characteristics--namely when the gap width is enhanced. PMID- 19219150 TI - Generation of ultrafast terahertz radiation pulses on metallic nanostructured surfaces. AB - A resonant "incoherent" rectification process is presented relying on the excitation of surface plasmons on a nanostructured metal surface. Excitation of gold and silver films with 800-nm femtosecond laser pulses results in the emission of terahertz radiation with an angle-dependent efficiency and an approximately third-order power dependence. It is shown that the source of this terahertz pulse generation is the surface-plasmon-assisted multiphoton ionization and ponderomotive acceleration in the evanescent field of the surface plasmon. Simple models are used to understand the forces and dynamics near the surface. PMID- 19219151 TI - Strain and temperature sensitivities of an elliptical hollow-core photonic bandgap fiber based on Sagnac interferometer. AB - We fabricated an elliptical hollow-core photonic bandgap fiber (EC-PBGF) by controlling lateral tension in the hollow core region during the fiber drawing process. The absolute value of group modal birefringence becomes relatively high near the bandgap boundaries. We also experimentally measured the strain and temperature sensitivities of the fabricated EC-PBGF-based Sagnac loop interferometer. The strain and temperature sensitivities were very much dependent upon the wavelength. Moreover this PBGF-based interferometer can be a good sensor of physical parameters such as strain and temperature. PMID- 19219153 TI - Nanostructuring of organic-inorganic hybrid materials for distributed feedback laser resonators by two-photon polymerization. AB - With two-photon absorption induced polymerization arbitrary three dimensional nano- and microstructures can be patterned directly into photoresists. We report on the fabrication of a low threshold organic semiconductor distributed feedback laser using the technique of two-photon absorption induced polymerization. A surface grating with 400 nm periodicity and 40 nm height modulation was fabricated by two-photon absorption induced polymerization in the organic inorganic hybrid material ORMOCER. With structuring several stacked layers acting as a planar basis for the nanostructure microscopic substrate tilt can be compensated simply. This enabled us to uniformly nano-structure the surface grating over an area of 200 x 200 microm(2). PMID- 19219152 TI - Tunable liquid microlens arrays in electrode-less configuration and their accurate characterization by interference microscopy. AB - A special class of tunable liquid microlenses is presented here. The microlenses are generated by an electrowetting effect under an electrode-less configuration and they exhibit two different regimes that are named here as separated lens regime (SLR) and wave-like lens regime (WLR). The lens effect is induced by the pyroelectricity of polar dielectric crystals, as was proved in principle in a previous work by the same authors (S. Grilli et al., Opt. Express 16, 8084, 2008). Compared to that work, the improvements to the experimental set-up and procedure allow to reveal the two lens regimes which exhibit different optical properties. A digital holography technique is used to reconstruct the transmitted wavefront during focusing and a focal length variation in the millimetre range is observed. The tunability of such microlenses could be of great interest to the field of micro-optics thanks to the possibility to achieve focus tuning without moving parts and thus favouring the miniaturization of the optical systems. PMID- 19219154 TI - 80 kHz repetition rate high power fiber amplifier flat-top pulse pumped OPCPA based on BIB3O6. AB - We present a high peak power optical parametric chirped pulse amplifier (OPCPA) seeded by a cavity dumped Ti:Sapphire oscillator. A frequency doubled high power Ytterbium-doped fiber amplifier is pumping the device. Temporal synchronization of the pump pulses is done via soliton generation in a highly nonlinear photonic crystal fiber. This soliton is fiber amplified and spectrally filtered in several fiber amplifiers. A simple birefringent pulse shaper generates a flat-top temporal pump pulse profile. Direct amplification of these pulses in large mode area fibers without using a stretcher and compressor provides significantly reduced complexity. For the first time to our knowledge broadband amplification around 800 nm central wavelength is demonstrated in BIB(3)O(6) (BIBO) crystals. The stretched Ti:Sapphire oscillator pulses are amplified up to a pulse energy of 25 microJ. Recompression with a grating compressor yields 50.7 fs pulses with 16.2 microJ pulse energy. PMID- 19219155 TI - Full simulations of the apertureless scanning near field optical microscopy signal: achievable resolution and contrast. AB - We simulate apertureless near-field optical imaging and obtain phase and amplitude scans of structured substrates for elastic scattering. The solution of the three-dimensional Maxwell equations does not involve approximations and we include large tips and substrates, strong interaction, interferometric detection and demodulation at higher harmonics. Such modeling represents a significant step towards quantitative simulations and offers the attractive possibility to study the individual influence of each relevant experimental parameter. We typically obtain highly localized signatures of the interaction of the tip with gold inclusions, superposed on a slowly varying background signal. The relative importance of both contributions and the achievable lateral resolution are strongly dependent on the geometry and scanning conditions. The simulations show sensitivity mostly to the first nanometers of the sample and underline the importance of scanning near the sample and being careful with mechanical anharmonicities on the tip oscillation. They also help to determine the influence of oscillation amplitude and demodulation harmonic. PMID- 19219156 TI - Preliminary use of nematic liquid crystal adaptive optics with a 2.16-meter reflecting telescope. AB - A nematic liquid crystal adaptive optics system (NLC AOS) was assembled for a 2.16-m telescope to correct for atmospheric turbulence. LC AOS was designed and optimized with Zemax optical software. Second, an adaptive correction experiment was performed in the laboratory to test the performance of the NLC AOS. After the correction, the peak to valley (PV) and root mean square (RMS) of the wavefront were down to 0.2 lambda (lambda=633 nm) and 0.05 lambda, respectively. Finally, the star of Pollux (beta Gem) was tracked using the 2.16-m Reflecting Telescope, and real time correction of the atmospheric turbulence was performed with the NLC AOS. After the adaptive correction, the average PV and RMS of the wavefront were reduced from 11 lambda and 2.5 lambda to 2.3 lambda and 0.6 lambda, respectively. Although the intensity distribution of the beta Gem was converged and its peak was sharp, a halo still existed around the peak. These results indicated that the NLC AOS only partially corrected the vertical atmospheric turbulence. The limitations of our NLC AOS are discussed and some proposals are made. PMID- 19219157 TI - Optical excitation and detection of vapor bubbles around plasmonic nanoparticles. AB - Laser-induced generation of the vapor bubbles in water around plasmonic nanoparticles was experimentally studied with optical scattering methods. Nanoparticle-generated bubbles temporally and spatially localize laser-induced thermal field and also amplify optical scattering relatively to that of gold nanoparticles. Bubble lifetimes and threshold fluencies were determined as functions of the laser (pulse duration, fluence, inter-pulse interval), nanoparticle (size, shape, aggregation state) and sample chamber parameters so to optimize bubble generation around plasmonic nanoparticles. Nanoparticle-generated bubbles are suggested as nano-scaled optical sensors and sources of localized thermal and mechanical impact. PMID- 19219158 TI - Efficient second harmonic generation from thin films of V-shaped benzo[b]thiophene based molecules. AB - We have designed an original approach for efficient Second Harmonic Generation of tailored V-shape benzo[b]thiophene molecular systems enabling versatile and flexible one-step, dry and technologically friendly thin film processing. The designed moieties show chi((2)) values at least as high as the reference LiNbO(3) single crystal, without poling processing and matching the constrains of integrated optical configuration for nonlinear optical devices. This may open the way to a new class of organic materials exploitable for photonic applications. PMID- 19219159 TI - Enhancement of ZnO photoluminescence by localized and propagating surface plasmons. AB - Insulating spacer layers of MgO were used to identify the enhancement mechanisms of the ZnO band-edge and visible luminescence in ZnO-MgO-Ag and ZnO-MgO-Au multilayers. Purcell enhancement of the ZnO band-edge emission by both Ag and Au surface plasmon polaritons is confirmed by demonstrating that the exponential decay of this emission as a function of increasing MgO thickness is consistent with the Ag and Au SPP evanescent decay lengths. Local surface plasmons excited in Ag and Au nanoparticles and rough films are also shown to enhance the ZnO visible donor-acceptor-pair photoluminescence by dipole-dipole scattering, again with an appropriate dependence on the thickness of the MgO spacer layer. We also confirm that both Ag and Au nanoparticles enhance the ZnO band-edge emission by charge transfer when the MgO spacer layer is absent. PMID- 19219160 TI - Fabrication and characterization of whispering-gallery-mode resonators made of polymers. AB - High-quality whispering-gallery-mode resonators made of polymethylmethacrylate (PMMA) are fabricated by simple mechanical turning and polishing according to a technique used by Ilchenko et al. to produce crystalline whispering-gallery-mode resonators with high quality factors (Q-factors). The high-Q PMMA resonators are investigated in two wavelength regimes: in the near infrared between the wavelengths 1470 and 1580 nm and at the wavelength 635 nm. The Q-factor in the infrared regime is limited by material absorption to 3 x 10(5) At 635 nm the Q factor is limited by surface scattering only and reaches 4 x 10(7), which is a new record for polymers. PMID- 19219161 TI - Diffraction Phase Cytometry: blood on a CD-ROM. AB - We demonstrate Diffraction Phase Cytometry (DPC) as a single shot, full-field, high throughput quantitative phase imaging modality, dedicated to analyzing whole blood smears. Utilizing a commercial CD as a sample substrate, along with dynamic spatial filtering via a liquid crystal spatial light modulator, we have developed a compact instrument capable of making quantitative, physiologically relevant measurements. To illustrate the ability of the system to function as a highly sensitive cytometer we imaged a large number (N=1,537) of live human erythrocytes in whole blood without preparation. We retrieved a comprehensive set of geometrical parameters including cell volume and surface area, which are not directly available using existing cytometers. Furthermore, we retrieved the minimum cylindrical diameter, through which red blood cells can pass, and deliver oxygen. These initial results prove the concept for an inexpensive lab-on-a-chip blood screening device. PMID- 19219163 TI - Macroscopic entanglement between a Bose Einstein condensate and a superconducting loop. AB - We theoretically study macroscopic entanglement between a magnetically trapped Bose-Einstein condensate and a superconducting loop. We treat the superconducting loop in a quantum superposition of two different flux states coupling with the magnetic trap to generate macroscopic entanglement. The scheme also provides a platform to investigate interferometry with an entangled Bose Einstein condensate and to explore physics at the quantum-classical interface. PMID- 19219162 TI - Design of transmission multilayer polarizer for soft X-ray using a merit function. AB - A new method of designing soft X-ray transmission multilayer polarizer for use at a single wavelength using a merit function has been eveloped. A merit function of product of p-transmittance throughput and logarithm of transmittance polarization ratio was chosen. Characteristics of Mo/Si multilayer calculated using the merit function at 13.0 nm have been compared with those calculated using the traditional method by the present authors and those reported so far. The merit function has given the most optimal results of throughput of 30.0% and polarization ratio of 202. The polarizers of much higher polarization ratio or much larger p-transmittance can be designed by choosing the number of layers and optimizing the thickness of each layer to maximize the merit function. Using this method, the roughness effect has been studied on Mo/Si and La/B multilayer polarizers at 13.0 nm and 6.7 nm, respectively. It was found that the influence of roughness is crucial in shorter wavelength region. PMID- 19219164 TI - Laser Doppler distance sensor using phase evaluation. AB - This paper presents a novel optical sensor which allows simultaneous measurements of axial position and tangential velocity of moving solid state objects. An extended laser Doppler velocimeter setup is used with two slightly tilted interference fringe systems. The distance to a solid state surface can be determined via a phase evaluation. The phase laser Doppler distance sensor offers a distance resolution of 150 nm and a total position uncertainty below 1 microm. Compared to conventional measurement techniques, such as triangulation, the distance resolution is independent of the lateral surface velocity. This advantage enables precise distance and shape measurements of fast rotating surfaces. PMID- 19219165 TI - Low loss high index contrast nanoimprinted polysiloxane waveguides. AB - Nanoimprint lithography is gaining rapid acceptance in fields as diverse as microelectronics and microfluidics due to its simplicity high resolution and low cost. These properties are critically important for the fabrication of photonic devices, where cost is often the major inhibiting deployment factor for high volume applications. We report here on the use of nanoimprint technology to fabricate low loss broadband high index contrast waveguides in a Polysiloxane polymer system for the first time. PMID- 19219166 TI - Effect of Wood's anomalies on the profile of extraordinary transmission spectra through metal periodic arrays of rectangular subwavelength holes with different aspect ratio. AB - The extraordinary transmission through silver film perforated with rectangular hole array with different aspect ratio was investigated. It was found that when the aspect ratio exceeded 7, the propagating surface plasmon polaritons (SPPs) transformed to localized resonance mode. The role of the Wood's anomaly on the shape of the transmission spectrum is investigated. By designing the rectangular hole arrays in a rectangular lattice, the Wood's anomaly can be shifted far apart from the transmission peak, the real localized resonance peak wavelength was identified and fitted well with the theoretical calculation using a simplified transmission-line model. PMID- 19219167 TI - Integrated optical modulator for signal up-conversion over radio-on-fiber link. AB - An integrated optical modulator, which consists of a dual-sideband suppressed carrier (DSB-SC) modulator cascaded with a single-sideband (SSB) modulator, is proposed for signal up-conversion over Radio-on-Fiber. Utilizing a single-drive domain inverted structure in both modulators, balanced modulations were obtained without complicated radio frequency (RF) driving circuits and delicate RF phase adjustments. Intermediate frequency (IF) band signal was up-conversed to 60GHz band by using the fabricated device and was transmitted over optical fiber. Experiment results show that the proposed device enables millimeter wave generation and signal transmission without any power penalty caused by chromatic dispersion. PMID- 19219168 TI - Suspended nanowires: fabrication, design and characterization of fibers with nanoscale cores. AB - We report a new approach for the fabrication of nanowires: the direct drawing of optical fibers with air suspended nanoscale cores. The fibers were made from lead silicate glass using the extrusion technique for preform and jacket tube fabrication. Fibers with core diameters in the range of 420-720 nm and practical outer diameters of 110-200 microm were produced, the smallest core sizes produced to date within optical fibers without tapering. We explored the impact of the core size on the effective mode area and propagation loss of these suspended nanowires relative to circular nanowires reported to date. As for circular nanowires, the propagation loss of these suspended nanowires is dominated by surface roughness induced scattering. PMID- 19219169 TI - The effect of Mie resonances on trapping in optical tweezers: comment. AB - Recently, Stilgoe, et al., [Opt. Express 16, 15039 (2008)] reported calculations of the force on an optically trapped sphere performed using the "optical tweezers toolbox". This software suffers from numerical inaccuracies that lead to qualitative errors in the state diagram for stable trapping, particularly for spheres smaller than the wavelength of light. PMID- 19219171 TI - Zero order synthetic hologram with a sinusoidal phase carrier for generation of multiple beams. AB - We discuss a phase synthetic hologram, for encoding arbitrary complex fields, whose design is based on a sinusoidal phase grating with a spatially modulated phase depth. An important feature of the hologram is that it encodes the complex field at the zero diffraction order of the carrier grating. The smoothness of this sinusoidal carrier grating facilitates the implementation of the hologram with a pixelated spatial light modulator. We take advantage of the hologram reconstruction at the zero-diffraction order for the simultaneous generation of a collection of complex beams. PMID- 19219172 TI - Measurement of coherent x-ray focused beams by phase retrieval with transverse translation diversity. AB - We describe a method for characterizing focused x-ray beams using phase retrieval, with diversity achieved by transversely translating a phase-shifting or absorbing structure close to the beam focus. The required measurements can be taken with an experimental setup that is similar to that already used for fluorescent scan testing. The far-field intensity pattern is measured for each position of the translating structure, and the collected measurements are jointly used to estimate the beam profile by using a nonlinear optimization gradient search algorithm. The capability to re-construct 1D and 2D beam foci is demonstrated through numerical simulations. PMID- 19219173 TI - Synthesized femtosecond laser pulse source for two-wavelength contouring with simultaneously recorded digital holograms. AB - A dual-wavelength femtosecond laser pulse source and its application for digital holographic single-shot contouring are presented. The synthesized laser source combines sub-picosecond time scales with a wide reconstruction range. A center wavelength distance of the two separated pulses of only 15 nm with a high contrast was demonstrated by spectral shaping of the 50 nm broad seed spectrum centered at 800 nm. Owing to the resulting synthetic wavelength, the scan depth range without phase ambiguity is extended to the 100-microm-range. Single-shot dual-wavelength imaging is achieved by using two CMOS cameras in a Twyman-Green interferometer, which is extended by a polarization encoding sequence to separate the holograms. The principle of the method is revealed, and experimental results concerning a single axis scanner mirror operating at a resonance frequency of 0.5 kHz are presented. Within the synthetic wavelength, the phase difference information of the object was unambiguously retrieved and the 3D-shape calculated. To the best of our knowledge, this is the first time that single-shot two-wavelength contouring on a sub-ps time scale is reported. PMID- 19219174 TI - Optical limiting in a periodic materials with relaxational nonlinearity. AB - We numerically investigate counter-propagating beams in a one-dimensionally, periodic structure with non-instantaneous Kerr nonlinearity for the design of efficient optical limiters. The performance of the Photonic Band Gap optical limiter with different response times is compared with the instantaneous case. Dynamic range and the cutoff intensity can be improved over a range of relaxation times. PMID- 19219175 TI - Effect of subwavelength annular aperture diameter on the nondiffracting region of generated Bessel beams. AB - A subwavelength annular aperture (SAA) made on metallic film and deposited on a glass substrate was fabricated by electron-beam lithography (EBL) and which was followed by a metal lift-off process to generate a long propagation range Bessel beam. We propose tuning the focal length and depth of focus (DOF) by changing the diameter of the SAA. We used finite-difference time domain (FDTD) simulations to verify our experimental data. We found that the position of the Bessel Beam focus spot (i.e. focal length) will be farther away from the SAA plane as the diameter of the SAA increases. In addition, the depth of focus (DOF) which is the length of the Bessel beam non-diffracting area, also increases as the diameter of the SAA expands. PMID- 19219176 TI - Plasmon enhanced luminescence from fullerene molecules excited by local electron tunneling. AB - Tunneling electrons from a scanning tunneling microscope (STM) induce luminescence from C(60) and C(70) molecules forming fullerene nanocrystals grown on ultrathin NaCl films on Au(111). Intramolecular fluorescence and phosphorescence associated with the transitions between the lowest electronic excited state and ground state of C(70) molecules are identified, leading to unambiguous chemical recognition on the nanoscale. Moreover we demonstrate that the molecular luminescence is selectively enhanced by localized surface plasmons in the STM tip-sample gap. PMID- 19219177 TI - Polymer photonic crystal dye lasers as Optofluidic Cell Sensors. AB - Dye doped hybrid polymer lasers are implemented as label free evanescent field biosensors for detection of cells. It is demonstrated that although the coverage is irregular and the cells extend over several lattice constants, the emission wavelength depends linearly on the fraction of the surface covered by the HeLa cells used as model system. Design parameters relating to photonic crystal sensing of large objects are identified and discussed. The lasers are chemically modified to bind cells and molecules with flexible UV activated linker molecules. PMID- 19219178 TI - Highly versatile fiber-based optical Fabry-Perot gas sensor. AB - We develop a versatile, compact, and sensitive fiber-based optical Fabry-Perot (FP) gas sensor. The sensor probe is composed of a silver layer and a vapor sensitive polymer layer that are sequentially deposited on the cleaved fiber endface, thus forming an FP cavity. The interference spectrum resulting from the reflected light at the silver-polymer and polymer-air interfaces changes when the polymer is exposed to gas analytes. This structure enables using any polymer regardless of the polymer refractive index (RI), which significantly enhances the sensor versatility. In experiments, we use polyethylene glycol (PEG) 400 (RI=1.465-1.469) and Norland Optical Adhesive (NOA) 81 (RI=1.53-1.56) as the gas sensing polymer and show drastically different sensor response to hexanol, methanol, and acetone. The estimated sensitivity for methanol vapor is 3.5 pm/ppm and 0.1 pm/ppm for PEG 400 and NOA 81, respectively, with a detection limit on the order of 1-10 ppm. Gas sensing for the analytes delivered in both continuous flow mode and pulsed mode is demonstrated. PMID- 19219179 TI - Properties of optical breakdown in BK7 glass induced by an extended-cavity femtosecond laser oscillator. AB - Using an extended-cavity femtosecond oscillator, we investigated optical breakdown in BK7 glass caused by the accumulated action of many laser pulses. By using a pump-probe experiment and collecting the transmitted pump along with the reflected pump and the broadband light generated by the optical breakdown, we measured the build-up time to optical breakdown as a function of the pulse energy, and we also observed the instability of the plasma due to the effect of defocusing and shielding created by the electron gas. The spectrum of the broadband light emitted by the optical breakdown and the origin of the material modification in BK7 glass was studied. We developed a simple model of electromagnetic wave propagation in plasma that is consistent with the observed behavior of the reflection, absorption, and transmission of the laser light. PMID- 19219180 TI - Maskless fabrication of nano-fluidic channels by two-photon absorption (TPA) polymerization of SU-8 on glass substrate. AB - We present a simple, fast, and repeatable method for fabricating nano-fluidic channels based on two-photon absorption (TPA) polymerization. Since our method does not require any mask, it is relatively cheaper and faster than other conventional lithography techniques. We illustrate that SU-8 has pronounced photoinitiation threshold behavior, which linearly decreases as the repetition rate increases. If the pulse energy and the repetition rate are controlled, channel width can be easily controlled. We report fluidic channels up to 110 nm in width, between polymerized parallel ribs, utilizing TPA and the photoinitiation threshold properties of SU-8. Finally, we also show that high repetition rate laser presents greater controllability in size of the polymerized region by varying fluence. PMID- 19219181 TI - Capacity of coherent free-space optical links using atmospheric compensation techniques. AB - We analyze the ergodic capacity and epsilon-outage capacity of coherent optical links through the turbulent atmosphere. We consider the effects of log-normal amplitude fluctuations and Gaussian phase fluctuations, in addition to local oscillator shot noise, for both passive receivers and those employing active modal compensation of wavefront phase distortion. We study the effect of various parameters, including the ratio of receiver aperture diameter to wavefront coherence diameter, the strength of the scintillation index, and the number of modes compensated. PMID- 19219182 TI - Imaging a vibrating object by Sideband Digital Holography. AB - We obtain quantitative measurements of the oscillation amplitude of vibrating objects by using sideband digital holography. The frequency sidebands on the light scattered by the object, shifted by n times the vibration frequency, are selectively detected by heterodyne holography, and images of the object are calculated for different orders n. Orders up to n = 120 have been observed, allowing the measurement of amplitudes of oscillation that are significantly larger than the optical wavelength. Using the positions of the zeros of intensity for each value of n, we reconstruct the shape of vibration the object. PMID- 19219183 TI - Compact beveled fiber optic probe design for enhanced depth discrimination in epithelial tissues. AB - We report the development and evaluation of a simple compact probe that incorporates multiple beveled fibers for depth sensitive detection of spectroscopic signals in vivo. We evaluated three probes with bevel angles 35, 40, and 45 degrees for their collection efficiency and depth resolution using a thin highly scattering white substrate and found that a 40 degree bevel provides the best characteristics for depth-resolved spectroscopy. The depth sensitivity of the probe with 40 degree beveled fibers was then evaluated using multilayer phantoms with scattering properties mimicking precancerous tissue and in vivo on normal human oral mucosa. The results demonstrate that the use of multiple beveled fibers has the capability to simultaneously collect scattering spectra from a range of depths within epithelial tissue that has the potential to provide further significant improvement of detection and monitorin of epithelial precancers. PMID- 19219184 TI - Large longitudinal electric fields (Ez) in silicon nanowire waveguides. AB - We demonstrate the presence of strong longitudinal electric fields (E(z)) in silicon nanowire waveguides through numerical computation. These waveguide fields can be engineered through choice of waveguide geometry to exhibit amplitudes as high as 97% that of the dominant transverse field component. We show even larger longitudinal fields created in free space by a terminated waveguide can become the dominant electric field component, and demonstrate E(z) has a large effect on waveguide nonlinearity. We discuss the possibility of controlling the strength and symmetry of E(z) using a dual waveguide design, and show that the resulting longitudinal field is sharply peaked beyond the diffraction limit. PMID- 19219185 TI - Diffuse reflectance spectroscopy detects increased hemoglobin concentration and decreased oxygenation during colon carcinogenesis from normal to malignant tumors. AB - Blood content and tumor oxygen level are important biomarkers and prognostic indicators in patients with colorectal cancer (CRC). However, noninvasive measurements of both quantities in human colon are limited. In this study, we extracted the total hemoglobin concentration (THC) and oxygen saturation (StO(2)) of normal, premalignant, and malignant colonic tissues in 27 patients using a diffuse reflectance instrument and algorithms based on the diffusion equation. The mean+/-standard error of THC and StO(2) from all normal sites (n=26) is 93.4+/-17.1microM and 67.2+/-3.7%, respectively. THC increased to 136.9+/ 23.8microM and 153.8+/-38.6microM and StO(2) decreased to 51.3+/-7.0% and 26.4+/ 6.1% for premalignant and malignant tissues, respectively. The disease-to-normal THC ratios are 3.2+/-1.1 and 4.4+/-1.9 and the disease-to-normal StO(2) ratios are 0.7+/-0.1 and 0.5+/-0.1 for pr alignant and malignant tissues, respectively. These results demonstrate the feasibility of a robust optical method to assess colon THC and StO2 at all stages of carcinogenesis in vivo so that the angiogenesis and hypoxia of the disease and the therapeutic role can be studied in CRC patients. PMID- 19219186 TI - Metal-insulator-metal nanorod arrays for subwavelength imaging. AB - We proposed using metal-insulator-metal (MIM) nanorod arrays for subwavelength imaging to improve the resolution based on the notable ability of field confinement of the MIM nanorods. The field distribution of a single Au-SiO(2)-Au nanorod, coupling between two nanorods and image transfer of shaped dipole sources through nanorod arrays were investigated by using the finite-difference time-domain method and compared with the case of Au nanorods. A resolution limit of approximately lambda/20 of the Au-SiO(2)-Au nanorod array was obtained, which is several times higher than that of the Au nanorod array. PMID- 19219187 TI - Effect of acoustic wave inertia and its implication to slow light via stimulated Brillouin scattering in an extended medium. AB - An analytical solution of the basic coupled stimulated Brillouin scattering (SBS) equations is obtained which describes the evolution of a Stokes pulse injected into an extended Brillouin medium excited by CW pump radiation. We show that regardless of the pulse length of the input Stokes pulse the delay of the output Stokes pulse results entirely from the inertia of the acoustic wave excitation via the electrostrictive interaction of the Stokes pulse with CW pump radiation. This delay is the SBS build up time and not the SBS induced group delay normally attributed to slow light. PMID- 19219188 TI - Analog mean-delay method for high-speed fluorescence lifetime measurement. AB - We present a new high-speed lifetime measurement scheme of analog mean-delay (AMD) method which is suitable for studying dynamical time-resolved spectroscopy and high-speed fluorescence lifetime imaging microscopy (FLIM). In our lifetime measurement method, the time-domain intensity signal of a fluorescence decay is acquired as an analog waveform. And the lifetime information is extracted from the mean temporal delay of the acquired signal. Since this method does not rely on the single-photon counting technique, the signals of multiple fluorescence photons can be acquired simultaneously. The measurement speed can be increased easily by raising the fluorescence intensity without a photon-rate limit. We have investigated various characteristics of our method in lifetime accuracy and precision as well as measurement speed. It has been found that our method can provide excellent measurement performances in various aspects. We hav demonstrated a high-speed measurement with a high photon detection rate of approximately 108 photons per second with a nearly shot noise-limited photon economy. A fluorescence lifetime of 3.2 ns was accurately determined with a standard deviation of 3% from the data acquired within 17.8 micros at a rate of 56,300 lifetime determinations per second. PMID- 19219189 TI - Frequency response in step index plastic optical fibers obtained from the generalized power flow equation. AB - We present a method to obtain the frequency response of step index (SI) plastic optical fibers (POFs) based on the power flow equation generalized to incorporate the temporal dimension where the fibre diffusion and attenuation are functions of the propagation angle. To solve this equation we propose a fast implementation of the finite-difference method in matrix form. Our method is validated by comparing model predictions to experimental data. In addition, the model provides the space time evolution of the angular power distribution when it is transmitted throughout the fibre which gives a detailed picture of the POFs capabilities for information transmission. Model predictions show that angular diffusion has a strong impact on temporal pulse widening with propagation. PMID- 19219190 TI - Limits for reduction of effective focal volume in multiple-beam light microscopy. AB - Employing interference patterns for illumination has been shown to reduce the focal volume in fluorescence microscopy. For example, the 4Pi technique employs two interfering laser beams and significantly decreases the focal volume, as compared to conventional microscopy. We study theoretically the effect of using multiple interfering laser beams on the focal volume. In realistic setups with three or four beams, the focal volume is about half of that from the 4Pi case. This improvement reaches a limit quickly as more beams are added, and for the idealized case of an infinite number of beams the focal volume is rather close to the three- or four-beam cases. Thus, our study suggests a limit for the possible reduction of the focal volume in a purely optical far-field setup. PMID- 19219191 TI - Metal nano-grid reflective wave plate. AB - We propose an optical wave plate using a metal nano-grid. The wave plate operates in reflection mode. A single-mode truncated modematching theory is presented as a general method to design such nano-grid wave plates with the desired phase difference between the reflected TM and TE polarizations. This analytical theory allows angled incidence calculations as well, and numerical results agree-well with comprehensive finite-difference time-domain electromagnetic simulations. Due to the subwavelength path-length, the reflective wave plate is expected to have improved broad-band functionality over existing zero-order transmissive wave plates, for which an example is provided. The proposed wave plate is simple and compact, and it is amenable to existing nanofabrication techniques. The reflective geometry is especially promising for applications including liquid crystal displays and laser feedback experiments. PMID- 19219192 TI - Three-dimensional harmonic holographic microcopy using nanoparticles as probes for cell imaging. AB - Luminescent markers play a key role in imaging techniques for life science since they provide a contrast mechanism between signal and background. We describe a new type of marker using second harmonic generation (SHG) from noncentrosymmetric BaTiO(3) nanocrystals. These nanoparticles are attractive due to their stable, non-saturating and coherent signal with a femtosecond-scale response time and broad flexibility in the choice of excitation wavelength. We obtained monodispersed BaTiO(3) nanoparticles in colloidal suspensions by coating the particle surface with amine groups. We characterized the SHG efficiency of 90-nm BaTiO(3) particles experimentally and theoretically. Moreover, we use the coherent SHG signal from BaTiO(3) nanoparticles for three-dimensional (3D) imaging without scanning. We built a harmonic holographic (H(2)) microscope which records digital holograms at the second harmonic frequency. For the first time, high-resolution 3D distributions of these SHG markers in mammalian cells are successfully captured and interpreted by the H(2) microscope. PMID- 19219193 TI - Nonlinear spectroscopy of cold atoms in diffuse laser light. AB - The nonlinear spectroscopy of cold atoms in the diffuse laser cooling system is studied in this paper. We present the theoretical models of the recoil-induced resonances (RIR) and the electromagnetically-induced absorption (EIA) of cold atoms in diffuse laser light, and show their signals in an experiment of cooling (87)Rb atomic vapor in an integrating sphere. The theoretical results are in good agreement with the experimental ones when the light intensity distribution in the integrating sphere is considered. The differences between nonlinear spectra of cold atoms in the diffuse laser light and in the optical molasses are also discussed. PMID- 19219194 TI - High sensitivity and large field enhancement of symmetry broken Au nanorings: effect of multipolar plasmon resonance and propagation. AB - The multipolar plasmon resonance and propagation of Au nanorings with symmetry broken were analyzed by using DDA and FDTD methods. Based on the multipolar plasmon resonance and propagation, we proposed ring-nanosensors with high sensitivities and optical ringnanoantennas with large local field enhancements. We revealed that the refractive index sensitivities of split nanorings are about 100% larger than those of perfect nanorings with same size; the local field intensity enhancement of split nanoring with three gaps has increased by 37% than that of dipole antennas. PMID- 19219195 TI - Multi-format receiver for non-return-to-zero binary-phase-shift-keyed and non return-to-zero amplitude-shit-keyed signals. AB - A Multi-format receiver for both non-return-to-zero binary-phase- shift-keyed (NRZ-BPSK) signal and non-return-to-zero amplitude-shift- keyed (NRZ-ASK) signal is demonstrated. Multi-format signal detection is based on incoherent BPSK demodulation and ASK-BPSK format conversion. Incoherent BPSK demodulation is realized by a Mach- Zehnder delay interferometer (MZDI) and a feedback decoder. Transmission experiments validate the feasibility of multi-format receiver. This receiver has potential to serve as a useful terminal block for all-optical wavelength division-multiplexed (WDM) networks. PMID- 19219196 TI - Analysis of coupling efficiency on hemispherical fiber lens by method of lines. AB - The method of lines (MoL) has been developed to study coupling efficiency on hemispherical lens. In this paper, the physical shape of the lens is approximated by cascading a number of straight waveguide segments. The perfectly matched layer (PML) is applied as an absorber for the MoL to reduce numerical reflection in the simulation region. Analysis is done by calculating coupling efficiency at the plane of integration where the coupling efficiency is an overlap integral between laser diode field and fiber field. The result of coupling efficiency in this analysis is compared to the experiment and ABCD matrix. It is found that MoL gives good result accuracy. PMID- 19219197 TI - Three dimensional digital holographic profiling of micro-fibers. AB - A method to measure the size, orientation, and location of opaque micro-fibers using digital holography is presented. The method involves the recording of a digital hologram followed by reconstruction at different depths. A novel combination of automated image analysis and statistical techniques, applied on the intensity of reconstructed digital holograms is used to accurately determine the characteristics of the micro-fibers. The performance of the proposed method is verified with a single fiber of known length and orientation. The potential of the method for measurement of fiber length is further demonstrated through its application to a suspension of fibers in a liquid medium. PMID- 19219198 TI - Slow light enhancement of nonlinear effects in silicon engineered photonic crystal waveguides. AB - We report nonlinear measurements on 80microm silicon photonic crystal waveguides that are designed to support dispersionless slow light with group velocities between c/20 and c/50. By launching picoseconds pulses into the waveguides and comparing their output spectral signatures, we show how self phase modulation induced spectral broadening is enhanced due to slow light. Comparison of the measurements and numerical simulations of the pulse propagation elucidates the contribution of the various effects that determine the output pulse shape and the waveguide transfer function. In particular, both experimental and simulated results highlight the significant role of two photon absorption and free carriers in the silicon waveguides and their reinforcement in the slow light regime. PMID- 19219199 TI - Quantum-rod dispersed photopolymers for multi-dimensional photonic applications. AB - Nanocrystal quantum rods (QRs) have been identified as an important potential key to future photonic devices because of their unique two-photon (2P) excitation, large 2P absorption cross section and polarization sensitivity. 2P excitation in a conventional solid photosensitive medium has driven all-optical devices towards three-dimensional (3D) platform architectures such as 3D photonic crystals, optical circuits and optical memory. The development of a QR-sensitized medium should allow for a polarization-dependent change in refractive index. Such a localized polarization control inside the focus can confine the light not only in 3D but also in additional polarization domain. Here we report on the first 2P absorption excitation of QR-dispersed photopolymers and its application to the fabrication of polarization switched waveguides, multi-dimensional optical patterning and optical memory. This fabrication was achieved by a 2P excited energy transfer process between QRs and azo dyes which facilitated 3D localized polarization sensitivity resulting in the control of light in four dimensions. PMID- 19219201 TI - Dielectric-loaded plasmonic waveguide-ring resonators. AB - Using near-field microscopy, the performance of dielectric-loaded plasmonic waveguide-ring resonators (WRRs) operating at telecom wavelengths is investigated for various waveguide-ring separations. It is demonstrated that compact (footprint approximately 150 microm(2)) and efficient (extinction ratio approximately 13 dB) WRR-based filters can be realized using UV-lithography. The WRR wavelength responses measured and calculated using the effective-index method are found in good agreement. PMID- 19219200 TI - Low bend loss in tightly-bent fibers through adiabatic bend transitions. AB - We demonstrate low bend loss for tightly bent optical fibers by winding the fiber around a mandrel designed to follow an adiabatic transition path into the bend. Light in the fundamental core-guided mode is smoothly transferred to a single cladding mode of the bent fiber, and back to the core mode as it leaves the bent region again. Design of the transition is based on modeling of the propagation and coupling characteristics of the core and cladding modes, which clearly illustrate the physical processes involved. PMID- 19219202 TI - Lasing with well-defined cavity modes in dye-infiltrated silica inverse opals. AB - Lasing in dye solution-embedded inverse silica opal structures was investigated. The opal films were prepared by sedimentation of polystyrene microspheres on a cover glass. The polystyrene structures were inverted using sol-gel infiltration of silica and subsequent removal of polystyrene. Photoluminescence of rhodamine (rhodamine B, 6G and sulfo-rhodamine 101) dye solutions embedded into the inverse silica opal structures exhibited clear signatures of the lasing via a distributed feedback (DFB) and gain modulation. The refractive index contrast between the dye and the inverse opal was small enough (approximately 0.03%) for the formation of refractive index coupling between the lasing modes. The lasing spectrum exhibited a highly regular periodic structure of modal peaks, rather than the chaotic superposition of peaks reported in previous studies. Lasing modes having a spectral width of about 0.25 nm and a free spectral range of about 0.75 nm appeared at the position of the maximum gain (the maximum fluorescence of the dye). PMID- 19219203 TI - Optimally chirped multimodal CARS microscopy based on a single Ti:sapphire oscillator. AB - We demonstrate high performance coherent anti-Stokes Raman scattering (CARS) microscopy of live cells and tissues with user-variable spectral resolution and broad Raman tunability (2500 - 4100 cm(-1)), using a femtosecond Ti:Sapphire pump and photonic crystal fiber output for the broadband synchronized Stokes pulse. Spectral chirp of the fs laser pulses was a user-variable parameter for optimization in a spectral focusing implementation of multimodal CARS microscopy. High signal-to-noise, high contrast multimodal imaging of live cells and tissues was achieved with pixel dwell times of 2-8 micros and low laser powers (< 30 mW total). PMID- 19219204 TI - Propagation of designer surface plasmons in structured conductor surfaces with parabolic gradient index. AB - In this work, we investigate the propagation of designer surface plasmons in planar perfect electric conductor structures that are subject to a parabolic graded-index distribution. A three-dimensional, fully vectorial finite-difference time-domain method was used to engineer a structure with a parabolic effective group index by modulating the dielectric constant of the structure's square holes. Using this structure in our simulations, the lateral confinement of propagating designer surface plasmons is demonstrated. Focusing, collimation and waveguiding of designer plasmons in the lateral direction is realized by changing the width of the source beam. Our findings contribute to applications of designer surface plasmons that require energy concentration, diffusion, guiding, and beam aperture modification within planar perfect electric conductors. . PMID- 19219205 TI - Observation and analysis of coherent optical waves emitted from large-Fresnel number degenerate cavities. AB - We demonstrate that the coherent optical waves emitted from large-Fresnel-number degenerate cavities persistently display a sort of salient intensity variations. We use the representation of the coherent states to explore the origin of the salient intensity variations and find that these coherent laser waves arise from a quadrature superposition of two degenerate coherent states. With the analytical representation of the superposed coherent states, we verify that these coherent laser waves possess a large angular momentum per photon. PMID- 19219206 TI - Hypocholesterolemia in clinically serious conditions--review. AB - BACKGROUND: Cholesterol is an essential component of cell membranes, precursor of steroids, biliary acids and other components of serious importance in live organism. Cholesterol synthesis is a complicated and energy-demanding process. Real daily need of cholesterol and mechanisms of decline cholesterol levels in critical ill are unknown. During stressful situations a significant hypocholesterolaemia may be found. Hypocholesterolemia has been known for a number of years to be a significant prognostic indicator of increased morbidity and mortality connected with a whole spectrum of pathological conditions. The aim of article is the elucidation of the role and importance of hypocholesterolaemia during the intensive care. METHODS AND RESULTS: We examined studies that are engaged in problems of hypocholesterolemia in critically ill. Very low levels of total as well as LDL cholesterol are most frequently found in serious polytrauma, after extensive surgery, in serious infections, in protracted hypovolemic shock. It is still not clear whether hypocholesterolemia reflects only a serious metabolic disorder, which results from a life-threatening condition, or whether it has an active role in evolution and outcome. CONCLUSIONS: Hypocholesterolemia is commonly observed in critically ill patients. Nevertheless, it is not known whether it is a secondary manifestation of disease, or whether it actively contributes to deterioration of the disease. Although the contribution of hypocholesterolemia to mortality is modest compared with known risk factors such as increased severity of illness and the development of nosocomial infection, low serum lipid concentrations represent a potential therapeutic target in sepsis. PMID- 19219207 TI - Current knowledge of methicillin-resistant Staphylococcus aureus and community associated methicillin-resistant Staphylococcus aureus. AB - BACKGROUND: Bacterial strains that are oxacillin and methicillin-resistant, historically termed methicillin-resistant Staphylococcus aureus (MRSA) are resistant to all beta-lactam agents, including cephalosporins and carbapenems. MRSA are pathogenic and have a number of virulence factors that enable them to result in disease. They are transmissible and important causes of nosocomial infections worldwide. An MRSA outbreak can occur when one strain is transmitted to other patients or through close contacts of infected persons in the community. Hospital-associated MRSA (HA-MRSA) isolates are also frequent causes of healthcare-associated bloodstream and catheter-related infections. Community associated MRSA (CA-MRSA) isolates are often only resistant to beta-lactam agents and erythromycin but they are an emerging cause of community-associated infections, especially skin and soft tissue infections (SSTI) and necrotizing pneumonia. METHODS: Current possibilities for detecting MRSA strains in the laboratory are reviewed and discussed in the context of the recent literature. RESULTS AND CONCLUSION: The active surveillance and prevention of MRSA occurrence and spreading in hospitals are discussed in the context of recent literature. PMID- 19219208 TI - Biodiversity of the chemical constituents in the epiphytic lichenized ascomycete Ramalina lacera grown on difference substrates Crataegus sinaicus, Pinus halepensis, and Quercus calliprinos. AB - AIM: The identification and evaluation of lichen metabolite production by the epiphytic lichenized ascomycete Ramalina lacera collected from different substrates: Crataegus sinaicus, Pinus halepensis, and Quercus calliprinos. METHODS: Chemical constituents were characterized by GC MS, HPLC, HR TLC, and other chemical methods. RESULTS: The most abundant fatty acids were alpha linolenic acid, oleic acid, and palmitic acid but a considerable variability of the ester composition from one to another was found. A comparison of neutral lipids, glycolipids, polar lipids and fatty acid composition of the tree growing lichen Ramalina lacera was done. Diacylglyceryl N,N,N trimethylhomoserine, diaclyglycerylhydroxymethyl N,N,N trimethyl beta alanine, phosphatidylcholine, and phosphatidylinositol were found as major components among polar lipids. Diffractaic, lecanoric, norstictic, protocetric, and usnic acids were isolated as major aromatic compounds in all samples of R. lacera. CONCLUSIONS: We evaluated a diversity of fatty acids, lipids, and aromatic compounds produced by the samples of Ramalina lacera growing on different tree substrates, Crataegus sinaicus, Pinus halepensis and Quercus calliprinos. PMID- 19219209 TI - Ascaridole and related peroxides from the genus Chenopodium. AB - AIM: Detection of monoterpenoid ascaridole and other terpenoids in the genus Chenopodium from the East Mediterranean. METHOD: Distribution of ascaridole in leaves of 13 species medicinal plant belonging to the genus Chenopodium was examined with the help of the GC MS method. RESULTS: cis Ascaridole was found as a major peroxy monoterpenoid (up to 46.9 %) in the oil. Three minor isomers: cis isoascaridole (1.1 6.4 %), trans ascaridole (1 2 %), and trans isoascaridole (1 2 %) were also detected. The biological activities and biosynthesis of ascaridole are further discussed. CONCLUSIONS: The results on Ascaridol and the major volatiles and semi volatiles of wild species belonging to the genus Chenopodium provide important information on biologically active monoterpenoid compounds and volatile metabolites biosynthesized in wild medicinal plants growing in the East Mediterranean. PMID- 19219210 TI - Chemerin is an independent marker of the metabolic syndrome in a Caucasian population--a pilot study. AB - AIM: Chemerin is a novel adipokine that has been suggested to play an important role in the pathogenesis of the metabolic syndrome. The aim of our study was to evaluate serum chemerin as a marker of the metabolic syndrome and to assess its predictive accuracy in a Caucasian population. METHODS: The study was designed as a cross-sectional study. Anthropometric measurements and serum analyses were done for Body Mass Index, waist circumference, chemerin, insulin, triacylglycerides, total cholesterol, HDL-cholesterol, LDL-cholesterol, uric acid, and glucose in 55 non-obese healthy subjects and 181 subjects at risk for the metabolic syndrome. ROC curves were determined and the Chi-squared test was used to analyse the data. RESULTS: Compared with healthy controls, subjects with suspected metabolic syndrome had significantly higher chemerin serum levels (medians: 266.0 vs.192.5 microg/l; P < 0.01). After further chemerin adjustment, the difference between the subgroups persisted. Chemerin serum levels correlated with age (r = 0.23), serum glucose (r = 0.23), HDL-cholesterol (r = -0.19), triacylglycerides (r = 0.22), systolic and diastolic blood pressure (r = 0.40; r = 0.24) and the number of metabolic syndrome risk factors (r = 0.47). At a serum chemerin cut-off level of 240 microg/l, the presence of the metabolic syndrome was diagnosed with 75 % sensitivity and 67 % specificity. CONCLUSION: In conclusion, serum chemerin levels are associated with the characteristics of the metabolic syndrome and could be an independent marker of this disorder in a Caucasian population. PMID- 19219211 TI - Pictorial cognitive task resolution and dynamics of event-related potentials. AB - AIMS: To judge whether and how the character of the visual stimulus and type of cognitive task affects brain event-related potentials (ERPs). METHODS: ERPs to three types of visual stimuli (white blank oval on a dark background, unfolded cube and net of sixteen small squares) were recorded from nine scalp sites and saved on a computer. Special software was used for off-line analysis of the ERPs. RESULTS: The presentation of each of the three visual stimuli used was followed by ERPs consisting of two negative (N160, N340) and one positive (P220) components. The character of the stimulus did not affect the latency of ERPs components. However, the type of visual stimuli affected the amplitude. The most conspicuous changes were shown by the N340 ERPs component. Its average amplitude in comparison with reference amplitude was always significantly higher during the first cognitive task ("Choose the cube that can be folded up from the unfolded cube!") and significantly lower than reference amplitude during the second cognitive task ("Complete the missing part of a figure with the appropriate item!"). It was also shown that subjective personality traits such as nervousness, spontaneous aggressivity and emotional lability had an influence on the recovery phase of the experiment affecting the average amplitude of N340 CONCLUSION: The results revealed that the cognitive processes underlying successful resolution of two pictorial cognitive tasks affected differently the activity of systems giving rise to visual ERPs. PMID- 19219212 TI - Methodological aspects of attempts to trans-differentiate adult stem cells into embryonic-like cells in vitro. AB - AIMS: The aim of this research was to set up an in vitro system to trans differentiate haematopoietic stem cells (HSCs) into embryo-like stem cells in order to de-differentiate them. In this more naive state they should be cultivated more easily in order to augment them for consecutive differentiation and autologous transplantation for use in clinical practice. METHODS: Using the principle of the methodology of blastocyst injection, HSCs were co-cultivated with mouse embryonic stem cells (mES) with and without cell to cell contact. After co-cultivation HSCs were analyzed by flow-cytometry using haematopoietic markers (CD34, CD45, CD133) and embryonic stem cell markers (SSEA-4, Tra-1-60, Tra-1-81). RESULTS: No ES cell markers were detected on the former HSCs. A decrease in HSC marker intensity was the only finding. This implies that no de differentiation took place. CONCLUSIONS: We hypothesize that the unnatural situation of a mixture of two cell types originating in different species may have led to this outcome. To achieve our goal of in vitro de-differentiation we need to use a purely human culture system without animal additives. PMID- 19219213 TI - Morphology of in vitro expanded human muscle-derived stem cells. AB - BACKGROUND: Skeletal muscle contains populations of multipotent adult stem cells also referred to as muscle-derived stem cells. AIM: The main goal of this study was to isolate and culture human adult stem cells from skeletal muscle and characterize them. METHODS: Muscle-derived stem cells were isolated from biopsy specimens of femoral muscle. The cells were cultured in Dulbecco's modified Eagle's minimal essential medium supplemented with 10% fetal calf serum and gentamycin. When they reached confluence, they were sub-passaged up to the third passage. Cells from the last passage were prepared for TEM analysis. Production of alpha-actin and desmin was confirmed by histochemistry. Moreover, the phenotypic characterization was performed. RESULTS: Primary isolated muscle derived stem cells had a fibroblast-like shape. During subsequent passages they maintained this morphology. TEM analysis showed typical ultrastructural morphology of mesenchymal stem cells. They had large pale nuclei with a large amount of euchromatine. Nuclei were irregular with noticeable nucleoli. Dilated cisterns of rough endoplasmic reticulum were present in cytoplasm. In certain parts of the cytoplasm there were aggregates of granules of glycogen. The products of cells were actively secreted into the extracellular matrix. They expressed alpha-actin and desmin. The results of phenotypic characterization showed that almost all analyzed cells were CD13, CD34, CD56 positive and CD45 negative. Moreover, they did not express anti-human fibroblast surface protein. CONCLUSIONS: Muscle-derived stem cells exhibited typical characteristics typical for mesenchymal stem cells. After analysis of their differentiation potential they could be used in tissue engineering and regenerative medicine. PMID- 19219214 TI - Morphology and mechanical properties of the subrenal aorta in normotensive and hypertensive rats. AB - AIMS: This work focuses on the morphological and mechanical changes in the wall of the subrenal aorta in rats suffering from arterial hypertension and chronic renal failure induced by subtotal nephrectomy (NX). METHODS: The quantitative structural parameters were assessed using stereological methods. The mechanical properties were determined using uniaxial tensile tests. RESULTS: Morphological results showed no significant differences in the wall structure of NX rats ten days following the subtotal nephrectomy in comparison with the control animals. Pronounced structural changes appeared ten weeks after the nephrectomy. The area of the profile of the arterial lumen, the volume fraction of elastin, and the elastin lamellar number in the tunica media were significantly higher in the NX rats than in the control animals. The values of the volume fraction of the smooth muscle cells in the tunica media and the lamellar unit thickness were significantly lower for the NX animals. Mechanical results showed that both kinds of tissues were characterized by a non-linear response when subjected to the tensile test. The moduli of elasticity of subrenal aortas in control and NX animals were different only for large deformations: NX samples had higher stiffness. The cyclic loading resulted in a time-dependent response for both tissue types. CONCLUSIONS: The results obtained from the NX rats ten days as well as ten weeks after operation suggested an outward hypertrophy of the aorta. The subrenal aortas of the NX animals had less strength and were less extensible than those of the control animals. PMID- 19219215 TI - Correlation of the IgG index and oligoclonal bands in the cerebrospinal fluid of patients with multiple sclerosis. AB - AIMS: The aim of this study was to assess the correlation between IgG index values and the number of the oligoclonal IgG bands (OCB) in the cerebrospinal fluid (CSF) of patients with multiple sclerosis (MS). MATERIAL AND METHODS: The set of 150 patients consisted of 41 males and 109 females (aged 18-68, mean 36.6 +/- 10.1 years). The CSF collected by a lumbar puncture was examined evaluating intrathecal synthesis using the IgG index and determining OCB. The number of alkaline OCB in the CSF was assessed using the method of isoelectric focusing. Pearson's correlation analysis, and homogeneity chi(2) test, Mann-Whitney test, paired-sample t-test (parametric) and Wilcoxon signed-ranks test (nonparametric) were used to evaluate the statistical significance of the results. RESULTS: No positive correlation between the IgG index and the number of OCB was found. Mann Whitney test also failed to demonstrate any significant difference of the IgG index values in patients both with the OCB number > or = 2 and < 2. CONCLUSION: This study did not confirm any correlation between the IgG index values and the OCB number in the CSF of MS patients. PMID- 19219216 TI - Transcranial sonography in movement disorders. AB - BACKGROUND: Transcranial sonography (TCS) in the B-mode has the ability to image, infratentorial and supratentorial brain structures. For this reason, it has potential use in the diagnosis and differential diagnosis of various intracranial pathologies. METHODS AND RESULTS: The authors reviewed the contribution of TCS to the differentiation of a number of neurodegenerative diseases: in parkinsonian syndromes, TCS can evaluate echogenicity changes in specific structures such as the hyperechogenic area of the substantia nigra (SN) in Parkinson's disease and the hyperechogenic caudate nucleus in Huntington's disease as well as the hyperechogenic lentiform nucleus (LN) in dystonia and Wilson's disease. In parkinson-plus syndromes, TCS may detect changes in width of the third ventricle and of the frontal horns of the lateral ventricle. The hyperechogenic SN can also be used in healthy populations as a marker of subclinical injury to the nigrostriatal system. CONCLUSION: TCS is a quick, safe and non-invasive method. It could be helpful in differentiation between several movement disorders together with clinical examination and other neuroimaging methods. PMID- 19219217 TI - Long term follow-up of neoadjuvant-adjuvant combination treatment of IIIA stage non-small-cell-lung cancer: results of neoadjuvant carboplatin/vinorelbine and carboplatin/paclitaxel regimens combined with selective adjuvant chemotherapy according to in-vitro chemoresistance test. AB - AIM: A prospective study investigated survival of patients with stage IIIA non small-cell-lung cancer (NSCLC) treated with a combination of neoadjuvant and adjuvant chemotherapy. METHODS: Consecutive chemo-naive patients with potentially operable stage IIIA NSCLC received carboplatin-based neoadjuvant treatment. Tumor cells harvested during surgery underwent methylthiazolyl tetrazolium blue (MTT) cytotoxic assay. After surgery, adjuvant chemotherapy was selected, where possible, according to MTT results. RESULTS: A total of 65 patients were evaluated (31 received carboplatin/vinorelbine, 34 carboplatin/paclitaxel). The overall response rate was 67.7 % (95% confidence interval [CI]: 56.3-79.1 %) with downstaging in 52.3 % (95% CI: 40.2-64.5 %) and no significant differences between regimens. Median follow-up was 86 months: median overall survival (OS) was 32.1 months (95% CI: 7.4-46.5), median time to progression was 25.1 months (95% CI: 15.1-34.9 months) and five-year overall survival was 35.7 % (95% CI: 23.7-47.7 %). Forty-seven patients (72.3 %) underwent surgery and 43 patients received adjuvant chemotherapy. Five-year survival after tumor resection was 49.5 % (95% CI: 34.2-64.8%), median OS was 59.0 months (95% CI: 34.2-83.1) and median disease free survival after surgery was 57.3 months (95% CI: 29.5-84.4). With MTT directed therapy, median OS was 85.1 months (95% CI: 15.4-148.6) and the 5-year survival rate was 57.0 % (95% CI: 34.5-79.5 %); the trend for longer survival failed to reach statistical significance. CONCLUSIONS: A combination of carboplatin-based neoadjuvant chemotherapy, surgical resection and adjuvant chemotherapy achieved satisfactory survival rates in stage IIIA NSCLC, especially in patients with complete resection of tumor and those given MTT-directed adjuvant treatment. Our results suggest MTT testing may help optimise adjuvant chemotherapy. PMID- 19219218 TI - Width of the great saphenous vein lumen in the groin and occurrence of significant reflux in the sapheno-femoral junction. AB - AIM: This study was performed to assess the contribution of the width of the anechogenous lumen of the great saphenous vein (GSV) in the groin measured by ultrasound (US) to the diagnostics of haemodynamically significant reflux (HSR) in the sapheno-femoral junction (SFJ). METHODS: We examined 200 lower limbs with primary varicose veins in 182 patients. Duplex scanning was performed with the patients in the supine position. Longitudinally imaging (B-mode) the inner anechogenous diameter of GSV was measured 4-5 cm distal to SFJ. PW Doppler sampling volume was placed at the same distance. The reflux was elicited by Valsalva manoeuvre. The HSR was defined as a backflow lasting > or = 1 s with a velocity of Vmax > or = 10 cm/s. RESULTS: The mean width of the GSV with HSR (n=152) was 6.39 mm, median 6.0 mm, SD +/- 2.21 mm. The mean width of GSV without HSR (n = 48) was 4.41 mm, median 4.4 mm, SD +/- 0.96 mm. The difference between the mean widths of GSV was statistically significant (p < 0.01). In GSV < 5 mm (n = 77) HSR (V max > or = 10 cm/s) was confirmed in 46 cases (59.7 %), in GSV > or = 5 mm (n = 123) in 106 cases (86.2 %). The sensitivity of dilatation of GSV > or = 5 mm for the presence of HSR in SFJ was 69.7 %, specificity 64.6 %. PPV (positive predictive value) of dilatation of GSV > or = 5 mm for the presence of HSR in SFJ was 86.2 %, NPV (negative predictive value) was 40.3 %, and the diagnostic accuracy of dilatation of GSV > or = 5 mm was 68.5 %. CONCLUSIONS: Measurement of anechogenous lumen GSV under the groin in B-mode is less sensitive (69.7 %) and less specific (64.6 %) in the diagnostics of HSR in SFJ. Only 68.5 % of all measurements of the width of the GSV below the groin in B-mode provided accurate indirect assessment of the functions of valves in SFJ. US scanning of the width of anechogenous lumen of GSV below the groin may serve only for ancillary examination. PMID- 19219219 TI - Spontaneous consolidation of scaphoid nonunion in a child. AB - BACKGROUND: Scaphoid nonunion is very rare in childhood. CASE REPORT: A 14 year old boy with scaphoid nonunion confirmed with CT did not past scheduled surgery. In spite of the short period of immobilisation union occurred and two years after the injury he was completely asymptomatic. CONCLUSIONS: We suspect that injury to the scaphoid, in this case may have accelerated the union shortly before the end of ossification. PMID- 19219220 TI - Influence of selected examination postures on shape of the spine and postural stability in humans. AB - AIMS: The accuracy of non-radiographic measurement of the spinal shape is influenced by postural sway (PS). The aim of this study was to determine whether certain examination postures prevent PS without changing key spinal characteristics. METHODS: We tested 1) natural standing position (posture A), 2) fixation postures standing with the support of the upper limbs against a wall (posture B) and 3) standing with the support of the head and chest against a wall (posture C). There were examined 60 subjects; the examination of each posture was repeated five times in each individual by DTP-3 microcomputer position system. The spinal shape changes resulting from tested postures were assessed using t test. The role of PS was assessed using standard deviations, and the significance of differences between the individual postures was determined using the F-test. RESULTS: Compared to posture A, the fixation posture B did not have any significant influence on the degree of PS, but it significantly influenced the shift of the spine from the ideal vertical and its slope, although this did not result in spine curve deformation. The fixation posture C significantly reduced the PS compared to posture A; however, there were significant changes in the spinal shape. This fixation posture shifted significantly the spine from the ideal vertical and slope characteristics. CONCLUSION: PS plays a role in the examination of spinal shape. Positions B and C did not meet the required criteria. Therefore, it is necessary to search for another examination position. PMID- 19219221 TI - Mistakes in dealing with aortic dissection. Lessons from three warning cases. AB - BACKGROUND: Aortic dissection is a dangerous condition with a high mortality in the acute stage. Aortic dissection requires early diagnosis and treatment. METHODS AND RESULTS: This short review discusses and focuses on known complications of aortic dissection and its natural mortality applying data from already published reports and from cohorts and registers, especially IRAD. Survival data of patients with type A and type B of dissection are presented and treatment options are proposed. The review presents three interesting cases from our database pointing out mistakes made in the diagnostic process and in dealing with the patient even after establishing the correct diagnosis. In one case, a patient with chest pain + "immeasurable" BP was suspected to suffer from an acute myocardial infarction and cardiogenic shock instead of AoD + aortic branch obstruction. In another patient with chest pain + V1V2 ST elevation, again the acute coronary syndrome was suspected. In fact, AoD with a perforation to cardiac chambers through the interventricular septum was the explanation. In the third case, the correct diagnosis of AoD was established. This patient was at a significant risk of aortic rupture because of his uncontrolled blood pressure. Instead of sedation administration and effective BP lowering, the patient was stressed even more by detailed information about this life threatening disease. This led to an aortic rupture with cardiac tamponade. Other mistakes made when dealing with all these presented cases are also discussed. CONCLUSION: The high mortality in patients suffering from aortic dissection is often potentiated by misdiagnosing and mishandling of these patients in clinical scenario. PMID- 19219222 TI - The choice of on- or off-pump coronary bypass grafting--a surgeon's dilemma. AB - BACKGROUND: Off-pump coronary bypass grafting (OPCAB) is a routine method of heart revascularization. This study was designed to compare the results of OPCAB and ONCAB performed by a single surgeon at our institution over a 3-year period. METHODS: The study enrolled 345 patients undergoing elective myocardial revascularization procedures, of which 173 (50.1%) were OPCAB and 172 (49.9 %) were ONCAB. The postoperative outcomes of both groups were analyzed, retrospectively. RESULTS: The mean number of grafts was significantly lower in the OPCAB group (2.9) than in the ONCAB group (3.7). There were no postoperative myocardial infarctions or hospital deaths in any of the patients. The postoperative total blood loss was significantly higher in the ONCAB group. The frequencies of postoperative confused states, strokes, postoperative transfusions, reoperations for bleeding, and atrial fibrillation, and the durations of intubation, ICU stay, and hospital stay did not differ significantly between the two groups. CONCLUSIONS: The OPCAB procedures are safe and provide good clinical outcomes, similar to those of ONCAB operations. The outcomes of this study favor neither of the compared techniques. PMID- 19219223 TI - Biliary manometric perfusion test in the evaluation of benign biliary stricture treatment--a case report. AB - BACKGROUND: Benign biliary strictures treated radiologically by percutaneous dilatation treatment are usually evaluated by 'clinical test'. For objective evaluation of treatment success, measurement of the pressure inside the biliary tree has been proposed. AIM: The aim of this article is to report our experience with the measurement of the pressure in the biliary tree after treatment, to present the biliary manometric perfusion test (BMPT) as an alternative method of treatment evaluation and to demonstrate why the perfusion test is a better method of evaluation. MATERIALS AND METHODS: A 65 year old lady with post operative biliary stricture and symptoms of jaundice was treated in our Department with percutaneous dilatation treatment and long-term drainage. The perfusion test (BMPT) was adopted for evaluating treatment success after the treatment for 9 months. An intrabiliary pressure less than 20 cms of water during perfusion test was considered success threshold. RESULTS: The test was carried out without any complications and the pressure inside the biliary tree was less than 20 cms of water during the test. Eventually the drain was removed. On follow-up at 6 months the patient developed no recurrence of jaundice and the quality of life improved clinically. Periodic liver function tests during follow-up were normal. CONCLUSION: The perfusion test (BMPT) is promising efficacious alternative method for the evaluation of radiological treatment of benign biliary strictures. It helps us to decide the end points in treatment and also helps in identifying patients who might later develop strictures. PMID- 19219224 TI - Chronic smoking and its effect on arterial stiffness. AB - AIMS: The goal of this study was to investigate the effect of chronic smoking on arterial stiffness at a peripheral site using pulse wave analysis. METHODS: Forty two non-smokers (17 males, 25 females) of average age 20.2 +/- 1.3 year and forty five smokers (19 males, 26 females) of average age 24.3 +/- 2.4 year were included in the study. Four parameters, SI (stiffness index), RI (reflection index), CT (crest time) and IWD (interwave distance) were evaluated by means of an adapted device based on pletysmographic principles that transform volume changes to voltage changes. RESULTS: SI corresponding to pulse wave velocity was 0.64 m/s higher in smokers than in non-smokers (7.25 +/- 0.53 m/s versus 7.89 +/- 0.73 m/s, P < 0.001). RI was significantly higher in smokers (42.49 +/- 6.71 %, versus 35.46 +/- 0.06 %, P < 0.001) than in non-smokers. IWD for non-smokers was 8.01 +/- 0.13 %, in smokers we found a 16 % increase to 9.21 +/- 0.83 % (P < 0.001). We detected a small increase in CT in smokers compared to non-smokers (0.09 +/- 0.01 s versus 0.10 +/- 0.01 s, P < 0.005). CONCLUSIONS: Chronic tobacco smoking is associated with endothelial dysfunction. In smokers we found increased values for all assessed parameters. Our results suggest that the negative effect of cigarette smoking on the vascular system can be found even in young smokers who have been smoking for less than 10 years. PMID- 19219225 TI - Survival analysis for clinical studies. AB - AIM: This paper focuses on the use of censored data in survival analysis. Survival analysis is used most frequently in the case of cancer patients when the study is finished and a number of individuals are still alive. The original article cited was declared recently to be the most cited statistical study in the biomedical area. The goal of this paper is to explain the basic principles and methods involved. The way survival analysis processes data and interprets outputs is presented using the clinical data of oncological patients. METHODS AND RESULTS: Survival analysis is used to estimate survivor function from survival data, to compare survivor functions and to assess the relationship of explanatory variables to survival time. These methods were applied to the data of 176 patients with hemato-oncological diagnoses who had undergone bone marrow blood transplant. CONCLUSION: It is very important to use appropriate methods when processing statistical data. Standard statistical procedures used for incomplete data could not provide the correct estimation. PMID- 19219226 TI - Bendable vs. angulated dental implants: consideration of elastic and plastic material properties based on experimental implant material data and FEA. AB - AIM: Bendable and angulated single-piece implants are used alternatively to screwable abutments in two-piece dental implant designs. Though used frequently, data on the stress distribution within such implants are not available and the question whether the bending contributes to fracture resistance has not been addressed. METHODS: We used the method of finite element to identify von Mises stresses and maximum stresses in bent and non-bent but angulated implants. Implants with one (e.g. applicable to screw designs) or two (applicable to basal implants) bending areas were the variables under investigation. RESULTS: For bends up to 13 degrees we discovered that if there is only one bend, the maximum stress is in the bent area. If two bends are made in two different bending areas, the maximum stresses are distributed between the two and, if either one of the bent areas is machined, there are no residual stresses within the implant body in this area. The maximum stresses are always located near the base-plates. The absolute value of the maximum stress is higher because no residual stresses are available to compensate stresses that stem from loading. CONCLUSION: Assuming that all other parameters are equal, bendable (basal) implants show a more even stress distribution along the vertical implant region than identically shaped implants with a machine-angulated area. Bendable basal implants therefore probably resist masticatory forces better than pre-angulated, machined implants, and unbent implants which provide a thin region in the vertical implant area. PMID- 19219227 TI - Practical highly enantioselective synthesis of (R)- and (S)-(E)-4-hydroxynon-2 enal. AB - Oxidative stress enhances lipid peroxidation (LPO) implicated in cancer promotion and progression. (E)-4-Hydroxynon-2-enal 1 (trans-4-hydroxy-2-nonenal, HNE) is one of the most abundant products of LPO. Reactions of HNE with DNA and proteins are responsible for its mutagenic and toxic effects. On the other hand, HNE is regarded as a key molecule in stress mediated cell cycle signaling. LPO generates racemic HNE (rac-1); however, it is expected that the individual enantiomers will behave differently in their interactions with cell components. The study of HNE stereochemistry in its chemical and biochemical interactions is hindered by the lack of expedient methods for preparation of pure enantiomers. This study presents one step synthesis of HNE in a cross-metathesis reaction between the commercially available oct-1-en-3-ol and acrolein in the presence of 2nd generation Grubbs catalyst. The use in the metathesis reaction of enantiomers of oct-1-en-3-ol obtained via Candida antarctica lipase resolution of the racemate allowed us to prepare of 4-(R)- and 4-(S)-enantiomers of HNE (R-1 and S-1, respectively) with excellent optical purity (97.5 and 98.4% ee, respectively) and good chemical yields (70%). PMID- 19219228 TI - Regulation of cell function by isoforms of C-reactive protein: a comparative analysis. AB - Despite the emerging evidence suggesting a proatherogenic role of C-reactive protein (CRP) in atherosclerosis, the contribution of CRP in pathogenesis of atherosclerosis and atherothrombosis has not been unequivocally defined. The role of CRP in pathophysiology/pathology seems to largely depend on its structure. Two CRP isoforms, the native pentameric and the modified monomeric one, differ substantially in their physiological functions, which is thought to origin from the considerable structural heterogeneity of the CRP molecule. The present review provides an overview of the experimental evidence with relevance to the clinical role(s) of various CRP isoforms. The biological role of the protein, its structure and distribution are discussed with particular emphasis on the diverse properties of the pentameric and monomeric forms of CRP. Some methodological aspects, related to experimental models and techniques of CRP preparation, are also critically reviewed. PMID- 19219229 TI - Purification and characterization of antibacterial proteins from granular hemocytes of Indian mud crab, Scylla serrata. AB - Marine invertebrates depend upon antimicrobial peptides (AMPs) as a major component of innate immunity, as they are rapidly synthesized and diffuse upon pathogen invasion. In this study, we report the identification and characterization of a 11 kDa antimicrobial protein, which we name SSAP (for Scylla serrata antimicrobial protein), from granular hemocytes of the mangrove crab S. serrata. The protein is highly similar to scygonadin, a male-specific AMP isolated from the ejaculatory duct of S. serrata. SSAP was isolated using various chromatographic techniques, viz. ion-exchange, ultra filtration and RP-HPLC, and demonstrated antibacterial activity against Gram positive and Gram negative bacteria. Full length mRNA encoding SSAP was amplified using a combination of RT PCR and RACE. The nucleotide sequence revealed a full-length ORF of 381 bp coding for a preprotein of 126 amino acids comprising a signal peptide of 24 amino acids and a mature protein of 102 amino acids with a predicted mass of 11435 Da and pI of 5.70. Unlike scygonadin, SSAP is expressed in several tissues of both male and female crabs, as evidenced by RT-PCR, Northern and Western blot analyses. The study suggests that SSAP might be an isoform or a variant of scygonadin and might play an important role in regulating the immunity of the crab upon microbial infection. PMID- 19219230 TI - Undergraduate studies in Public Health: key elements in an essential debate. PMID- 19219231 TI - [Cost-utility analysis as a mechanism for allocation of health care resources: a review of the debate]. AB - Priority setting in health care involves choosing between alternative health care programs and/or patients or groups of patients who will receive care. Traditionally, health economists have proposed maximizing the additional health gain measured in QALYs as a way of setting priorities and maximizing social welfare. This requires that the social value from health improvements be a product of gains in years of life, quality of life, and number of people treated. The results of a literature review suggest that potential health gain is not a single relevant determinant of value, nor is the rule of maximizing this gain sufficient. The social value of a health gain appears not to be linear in terms of mortality and morbidity, or neutral vis-a-vis people's characteristics or the ultimate distribution of health in society. In parallel with the review of the debate on the role and limitation of QALYs for prioritizing health care resources, the article attempts to justify the controversy over some empirical results, particularly in relation to the construction and expression of social preferences. PMID- 19219232 TI - [Validity of self-reported gestational diabetes mellitus in the immediate postpartum]. AB - Studies based on self-reporting of risk factors or morbidity are useful in public health, due mainly to the low cost and ease of data collection, but the accuracy of the resulting data is important for the internal validity of the research. The aim of this study was to evaluate the agreement between self-reported gestational diabetes mellitus and prenatal care medical records (gold standard). Agreement was measured by sensitivity, specificity, accuracy, kappa statistic, and confidence intervals (95%CI). From July to December 2007, all the women who gave birth in three maternity hospitals in Pelotas, Rio Grande do Sul, Brazil, were visited in the immediate postpartum. 1,047 women were interviewed, of whom 872 were in possession of their prenatal care card; of these, 869 had their blood glucose recorded on the card. Mean age was 26.7 years. Prevalence of gestational diabetes mellitus based on the gold standard was 4.3% (95%CI: 3.0-5.8), while the self-reported rate was 4% (95%CI: 2.8-5.5). Sensitivity of self-reported gestational diabetes mellitus was 72.9% (95%CI: 55.9-86.2); specificity was 99% (95%CI: 98.1-99.6); accuracy 97.9% (95%CI: 96.7-98.7); kappa 74%. For this population, self-reported gestational diabetes mellitus is valid information and can be used in population-based studies. PMID- 19219233 TI - [Development of secondary dental care: a study on specialized dental clinics in Pernambuco State, Brazil]. AB - This exploratory study aimed to evaluate the Specialized Dental Care Centers in Pernambuco State, Brazil, based on secondary outpatient treatment data from 2006 and standards and criteria adopted to implement these services. The study describes the overall achievement of targets by these specialized dental clinics and variables for characterization of the dental services and municipalities (counties). The methodology included descriptive analysis and non-parametric correlation analysis (Spearman coefficient), with significance set at 5%. Of the 22 specialized dental clinics, 40.9% showed good performance, and 81.8% met the target for basic dental care, as compared to 13.6% that met the target for minor oral surgery. Among the independent variables, only population size and Municipal Human Development Index (MHDI) were correlated with the performance of the specialized dental clinics: the smaller the city (r = 0.678; p < 0.001) and the lower the MHDI (r = 0.599; p < 0.001), the worse the performance. The findings suggest that standardization of the implementation and functioning of specialized dental clinics should be monitored and evaluated in order to ensure better quality of services for the population. PMID- 19219234 TI - [Multiple imputations for missing data: a simulation with epidemiological data]. AB - In situations with missing data, statistical analyses are usually limited to subjects with complete data. However, such estimates may be biased. The method of 'filling in' missing data is called imputation. This article aimed to present a multiple imputation method. From a data set of 470 surgical patients, logistic models were developed for death as the outcome. Two incomplete data sets were generated: one with 5% and another with 20% of missing data in a single variable. Logistic models were fitted for the complete and incomplete data sets and for the data set completed by multiple imputations. Estimates obtained for the data set with missing data were different from those observed in the complete data set, mainly in the situation with 20% of missing data. The multiple imputation used here appeared efficient, producing very similar results to those obtained with the complete data set. However, one coefficient became non-significant. The analysis using multiple imputations was considered superior to using the data sets that excluded incomplete cases from the analysis. PMID- 19219235 TI - [The role of the court system in regulating health insurance plans in Brazil]. AB - Consumer complaints against private health insurance plans and companies in Brazil have become increasingly frequent in the country's 'supplementary' (non public) health care sector, with numerous cases reaching the courts. The problem raised the need for regulation of this private market, which began in 1998, through Law no. 9.656. One of the challenges faced by the National Agency for Supplementary Health Care (ANS) is resistance to the legislation by health insurance companies, besides the fact that there are still some contracts not covered by this law. The objective of the current study was to analyze health insurance policyholders' appeals against court rulings for or against injunctions concerning coverage, in cases heard by the courts in Rio de Janeiro and Sao Paulo. The main data investigated were: court issuing the ruling; defendant; basis for the case; ruling by the Circuit Court and Court of Appeals; and the legal arguments. Based on the findings, the Brazilian court system still plays an important role in hearing and ruling on complaints by health insurance policyholders. The ANS has an important role in filling some gaps that have still not been solved in regulating the health insurance industry. PMID- 19219236 TI - [Effect of weekly rice fortification with iron on anemia prevalence and hemoglobin concentration among children attending public daycare centers in Rio de Janeiro, Brazil]. AB - This was a double-blind, placebo-controlled, community-based trial. For 16 weeks, children in the intervention group (n = 180) received iron-fortified rice, while children in the control group (n = 174) received rice with placebo. Anemia was defined as hemoglobin < 11.0g/dL. Student's t-test was used to compare mean variation in hemoglobin between the groups. Hemoglobin concentration improved in both groups, with a mean increase of 0.42g/ dL in the intervention group (11.28+/ 1.23g/dL to 11.75+/-1.16g/dL, p < 0.001), and 0.49g/dL in controls (11.06+/ 1.13g/dL to 11.51+/-1.16g/dL, p < 0.001). Anemia decreased (p < 0.01) in both groups (37.8% to 23.3% in the intervention group and 45.4% to 33.3% in controls), with no significant difference between the two. Hemoglobin increase was significantly higher in children that received total iron = 53.76mg from fortified rice, compared to those who received less than this cut-off value (0.94g/dl vs. 0.39g/dl, p = 0.03). The findings suggest that this type of intervention can be useful in anemia control as long as fortified food intake is adequate. PMID- 19219237 TI - [Main factors associated with motor vehicle accidents on Federal Highway 163, Mato Grosso State, Brazil, 2004]. AB - Transportation is essential to the economic development of Mato Grosso State, Brazil, and Federal Highway 163 is the main route for marketing regional agricultural produce. One unfortunate consequence of this activity is the occurrence of motor vehicle accidents. With the aim of analyzing and categorizing the main factors associated with motor vehicle accidents on this highway, the current study used data from the Mato Grosso State Division of the Brazilian Federal Highway Patrol for the year 2004. Factors significantly associated with accidents involving casualties were: substandard paving (OR = 1.89; CI: 1.32 2.70); front-end collisions (OR = 14.14; CI: 8.96-22.32); and running over pedestrians (OR = 35.95; CI: 8.10-159.52. Contributing factors included highway maintenance problems (OR = 4.35; CI: 1.94-9.75) and disobeying traffic signs (OR = 5.69; CI: 2.01-16.12). In conclusion, reduction of motor vehicle accidents on this highway requires comprehensive intervention rather than targeting measures only to drivers or highway patrol personnel, but considering regional issues and their economic, social, and cultural dimensions. PMID- 19219238 TI - Prescription errors in Brazilian hospitals: a multi-centre exploratory survey. AB - In Brazil, millions of prescriptions do not follow the legal requirements necessary to guarantee the correct dispensing and administration of medication. This multi-centre exploratory study aimed to analyze the appropriateness of prescriptions at four Brazilian hospitals and to identify possible errors caused by inadequacies. The sample consisted of 864 prescriptions obtained at hospital medical clinics in January 2003. Data was collected by three nurse researchers during one week using a standard data sheet that included items about: the type of prescription; legibility; completeness; use of abbreviations; existence of changes and erasures. There were statistically significant differences between incomplete electronic prescriptions at hospital A, and handwritten ones from hospitals C (C2 = 12.703 and p < 0.001) and D (C2 = 14.074 and p < 0.001). Abbreviations were used in more than 80% of prescriptions at hospitals B, C and D. Changes were found in prescriptions at all hospitals, with higher levels at hospitals B (35.2%) and A (25.3%). This study identified a range of vulnerable points in the prescription phase of the medication system at the hospitals. Physicians, pharmacists and nurses should therefore jointly propose strategies to avoid these prescription errors. PMID- 19219239 TI - [Mortality for oral cancer and socioeconomic status in Brazil]. AB - Chronic non-communicable diseases represent a major public health problem, requiring more effective investigation and control by government agencies. The aim of this study was to correlate the mortality rate for oral cancer in Brazilian State capitals from 1998 to 2002 with socioeconomic factors collected in the 2000 census, using an ecological study design. Data were obtained from the Mortality Information System from 1998 to 2002. Social factors were taken from the Brazilian Human Development Atlases. After data collection, statistical analysis was performed using Pearson's correlation index. The findings included positive and significant correlations among the socioeconomic indicators (Municipal Human Development Index-- MHDI, MHDI-income, MHDI-education, MHDI-life expectancy, and per capita income), and negative and significant correlations with the socioeconomic indicators Gini Index and infant mortality. Despite the study's limitations and probable underreporting in less developed State capitals, the study found significant statistic correlations between the selected socioeconomic indicators and the oral cancer mortality rate. PMID- 19219240 TI - [Demographic profile of Bororo Indians from Mato Grosso State, Brazil, 1993 1996]. AB - This paper analyzes the demographic profile of the Bororo Indians from Mato Grosso State, Brazil, from 1993 to 1996. Data came from annual collection and registration of vital statistics conducted by the health service in three villages (Garcas, Meruri, and Morada dos Bororo). The average annual population growth rate was 2.4%. Nearly half (44%) of the population was younger than 15 years (median 16 years). The crude birth rate was 30.9 per 1,000 and the crude death rate 7.3 per 1,000 inhabitants. The infant mortality rate was high (58.8 per thousand live births), probably resulting from precarious health conditions in the villages. The results showed that women married younger than men and that there were numerous unmarried adult men (26.7%) and women (13.3%), as compared to data from other indigenous groups. The total fertility rate of Bororo women was 4.3, quite low compared to other indigenous communities in Brazil. The study emphasizes the importance of systematically collecting and analyzing demographic data on indigenous populations. PMID- 19219241 TI - [Violence against children in the city of Ribeirao Preto, Sao Paulo State, Brazil: child abuse prevalence estimated from school system data]. AB - The official statistics on child abuse fail to reflect reality, since many cases are not reported. The current study aimed to estimate the real prevalence of child abuse in children 0 to 10 years of age enrolled in public and private daycare centers and schools in the city, using information obtained from teachers and staff, based on the premise that they are well-positioned to identify cases, and then to compare this estimated prevalence to the official rate. The instrument used was the Epidemiological Manual, and the random, representative sample consisted of 305 teachers, responsible for 6,907 children. The resulting prevalence was 5.7%, compared to 0.3% as calculated by data from the Tutorial Councils, thus corroborating the hypothesis that the official rate merely represents the tip of the iceberg. The most frequent forms of abuse identified by both the public and private schools were neglect and psychological abuse, generally considered less important, since they are viewed as less serious. The problems were also shown to become worse as the children grew older, indicating the importance of network collaboration for detection, notification, and early intervention in such situations, under the logic of secondary prevention. PMID- 19219242 TI - [Opinions by physicians from the Family Health Program on four health care priorities proposed by the Agenda for Commitment to Comprehensive Child Health and Reduction of Infant Mortality]. AB - The aim of this study was to understand how physicians from a team in the Brazilian Family Health Program perceive their work in child health, as compared to the program's principles outlined in the Agenda for Commitment to Comprehensive Child Health and Reduction of Infant Mortality, under the Ministry of Health. The backdrop was the strategy for implementation of primary care under the Unified National Health System (SUS). Semi-structured interviews were held, and the material was submitted to content analysis. According to the findings, in general the Agenda is being met. However, there are difficulties with referral and counter-referral; the measures proposed by the Family Health Program require medical and sociological competence and face problems inherent to biomedical training; the infrastructure and inputs are precarious; and the training provided does not prepare physicians sufficiently for a more comprehensive approach. In conclusion, the primary care model in Brazil requires adjustments to the country's reality, and partnerships that transcend the system are necessary. PMID- 19219243 TI - [Brazilian teenagers and beer advertising: relationship between exposure, positive response, and alcohol consumption]. AB - Brazilian teenagers report problematic patterns of alcohol consumption. Alcohol advertising strategies are one of the main factors influencing adolescents' alcohol consumption. The aim of this study was to evaluate the relationship between positive responses to TV beer commercials, exposure, and alcohol consumption. Thirty-two recent TV commercials were shown to 133 high school students from public schools in Sao Bernardo do Campo, Sao Paulo State, Brazil. The subjects recorded how well they liked the ads and how often they had already watched each commercial. The teenagers also reported their alcohol consumption rates. The ten commercials analyzed in this article were the five most popular and the five least popular. The analysis showed that subjects had already seen the five most popular ads, but not the five least popular. In addition, the five most popular ads received higher scores from teenagers that reported having consumed beer during the previous month. The study found a positive relationship between enjoying beer advertising and exposure to beer ads, as well as between alcohol consumption and positive responses to alcohol commercials. PMID- 19219244 TI - [Reliability and validity of death certificates specifying oral cancer as cause of death in Teresina, Piaui State, Brazil, 2004-2005]. AB - Brazilian health policy depends on the accuracy of data in the health information system. This study aimed to assess the reliability and validity of data on underlying cause of death due to a specific neoplasm in the Municipality of Teresina, Piaui State, Brazil, based on all the death certificates from 2004 and 2005 that reported oral cancer as the underlying cause of death (total of 23 death certificates). The death certificates were recoded for underlying cause of death by using a form targeted at collecting clinical and laboratory data to confirm the medical diagnosis on patient charts. Diagnosis was confirmed by histopathologic examination and clinical history. Simple agreement was 91.3%, kappa coefficient 0.84, and positive predictive value 90.9%. Based on the findings, the mortality statistics were valid and reliable. One limitation to this study was the possible underreporting of cases with oral cancer as the underlying cause of death, given that such data are not included in the mortality information system in Teresina. PMID- 19219245 TI - [Prevalence of high blood pressure and associated factors in students from public schools in Greater Metropolitan Recife, Pernambuco State, Brazil, 2006]. AB - The aim of this study was to identify the prevalence of systemic arterial hypertension (high blood pressure) and associated health factors in adolescents, using a cross-sectional, population-based, epidemiological design from April to September 2006, within a stratified sample according to school size and shift. Using the Portuguese version of the Global School-Based Student Health Survey questionnaire, 1,878 students from 29 public schools in Greater Metropolitan Recife, Pernambuco State, Brazil, were analyzed for: age, gender, body mass index, fruit, vegetable, alcohol, and tobacco consumption, as well as blood pressure, adopting the 1996 Task Force Report on High Blood Pressure in Children and Adolescents and Centers for Disease Control and Prevention parameters for nutritional status. Prevalence rates for high blood pressure, overweight, and obesity were 17.3%, 6.9%, and 3.7%, respectively. Factors associated with high blood pressure were: male gender, obesity, overweight, and lack of physical activity. Knowledge of factors associated with arterial hypertension in adolescents can help support health education campaigns. PMID- 19219246 TI - [Mapping harm reduction programs in Greater Metropolitan Porto Alegre, Rio Grande do Sul State, Brazil]. AB - This study mapped and described 11 harm reduction interventions/programs in Greater Metropolitan Porto Alegre, Rio Grande do Sul State, Brazil, in 2004-2006. Mapping was based on interviews and analysis of available documents and comparison with a previous study with data from 2003. We aimed to discuss the programs' sustainability (political, financial, and administrative) and operational characteristics, based on the following categories: type of links in the program or intervention; forms of inclusion in the municipalities; background, financing; typical activities; team hiring practices; volunteer work, if any; resources (human and financial); partnerships; municipal legislation, if any; and participation in forums for political representation. Despite the diversity of links and organizational formats, there were some common characteristics: precarious professional status; dependence on volunteer work to implement activities; influence of turnover in Municipal and State administration on the continuity of projects and partnerships, as well as on the maintenance of human and financial resources. We conclude that such factors lead to lack of continuity in the services provided to the target population. PMID- 19219247 TI - Oral carcinoma epidemiology in Parana State, Southern Brazil. AB - Oral carcinoma is the sixth most frequent type of cancer in the world and the seventh most common in Brazil (the country with the highest incidence in Latin America). Mean five-year survival remains one of the lowest among the main cancers, thus justifying studies that contribute to the development of preventive strategies. The aim of this study was to compare the epidemiological, clinical, and histological characteristics of 91 patients with oral carcinoma. Mean age was 58.62 +/- 10.46 years, and male-to-female ratio was 6.6:1.0 (79 men and 12 women). European descendants predominated with 79 patients (86.8%). Eighty-five individuals (93.4%) smoked and 70 (76.9%) consumed alcohol regularly. Anatomical distribution of tumors was: 27 (29.7%) tongue; 18 (19.8%) floor of mouth; 11 (12.1%) oropharynx; and 11 (12.1%) oral mucosa. Fifty-seven patients (62.6%) presented lymph node involvement and three (3.3%) had distant metastases. Surgery and radiotherapy were used in 43.2% of patients. With the exception of the male/female ratio (which was higher), our data are consistent with previous studies on oral carcinoma patients. PMID- 19219248 TI - [Analysis of data from Poison Control Centers in Rio de Janeiro, Brazil, for use in public health activities]. AB - Health information systems should be sources of reliable and valid data that can be used for surveillance to reduce morbidity and mortality. Unintentional poisoning is highly relevant to public health, since it is the most frequent cause of emergency pediatric care. Information on such events is recorded in Poison Control Centers (PCCs). The current study analyzed the records on unintentional poisoning with household cleaning products in two PCCs in the State of Rio de Janeiro, Brazil, in 2000-2002. The variables analyzed were gender, age, cause of the accident, exposure route, toxic agent, and outcome. In the 2,810 records studied, boys four years and younger were the most vulnerable group and oral ingestion was the most common exposure route. However, analysis of the data on the toxic agent, cause, and outcome showed that the data currently recorded in the PCCs are inconsistent. It is necessary to review the definitions used by the PCCs in order for the resulting data to effectively support public health measures and health surveillance policies. PMID- 19219249 TI - [Nutritional profile and factors associated with malnutrition in Kaingang children on the Mangueirinha Indigenous Reserve, Parana State, Brazil]. AB - The goal of this study was to evaluate the nutritional status of indigenous children and to investigate factors associated with nutritional deficits. Weight and height measurements were obtained for 141 Kaingang children from 0 to 5 years of age living on the Mangueirinha Indigenous Reserve in Parana State, Brazil. Data on maternal and infant conditions and socioeconomic characteristics were also gathered through face-to-face interviews. Based on World Health Organization criteria (2006), 24.8% of the children presented low height-for-age (HA), 9.2% low weight-for-age (WA), 2.1% low weight-for-height (WH), and 2.1% low weight according to body mass index for age (BMIA). Based on NCHS criteria (1977), 19.9% of the children presented low HA, 9.2% low WA, and 1.4% low WH. 6.4% were overweight according to BMIA. Low birth weight and non-masonry housing construction were associated statistically with nutritional deficits. The Kaingang children are subject to poor living conditions, associated with deficits in their nutritional profile. PMID- 19219250 TI - [Objective and subjective factors related to self-rated oral health among the elderly]. AB - The aim of this study was to investigate factors associated with negative self rated oral health. Elderly subjects from the Brazilian Ministry of Health's oral health survey (2002-2003) who rated their own oral health as bad or very bad were compared to those who rated their oral health as excellent, good, or fair, using prevalence ratios based on Poisson regression. The minority (870; 17%) gave their own oral health a negative rating. Negative self-rated oral health was less prevalent among those with 1-9 teeth and the edentulous, and more prevalent among blacks, mixed-race, and indigenous, those who had never used dental service, with soft tissue alterations, who reported little pain or medium to intense pain, rated their own appearance and chewing as fair or bad or very bad, reported that their oral health limited their social interaction a little or considerably, and reported needing dental treatment (PR = 1,47; 95%CI: 1.20-1.79). Despite their precarious oral conditions, the majority gave their oral health a positive rating. Subjective conditions were more heavily associated with self-rated oral health than were objective conditions. The findings suggest unequal oral health conditions and allow orienting public policies aimed at oral health and quality of life. PMID- 19219251 TI - [Coping strategies, functional difficulties, and associated factors in institutionalized elderly]. AB - The aim of this study was to analyze the strategies used by elderly people to cope with functional difficulties, and to investigate a possible association between such strategies and variables like depressive symptoms, gender, schooling, marital status, and self-perceived health. 103 institutionalized elderly individuals participated in the study. They required assistance for at least one activity of daily living. The study was cross-sectional and used the following indicators: Personal Data Chart, Activities of Daily Living Scale, Modified Mini International Neuropsychiatric Interview, Mini-Mental Examination, Coping Strategies Inventory, and Geriatric Depression Scale. The most widely used coping strategies in the sample were self-control and problem solving. Significant associations were found between schooling and depression. The findings suggest that assessing coping strategies for stressful events can assist analysis of the situation and appropriate adaptation of the chosen strategies, so as to foster changes within the medical context. PMID- 19219252 TI - [Neonatal screening program at the university hospital of the Ribeirao Preto School of Medicine, Sao Paulo University, Brazil]. AB - The Neonatal Screening Program at the University Hospital of the Ribeirao Preto School of Medicine, Sao Paulo University, Brazil, was introduced in 1994. As of December 2005, congenital hypothyroidism had been diagnosed in 76 infants, phenylketonuria in 10, and hemoglobinopathies in 25, representing incidence rates of 1:2,595, 1:19,409, and 1:4,120, respectively. A total of 2,747 newborns had the sickle cell trait, i.e., were heterozygous for the sickle mutation (1:37.5 live births). The program's mean coverage during this period was 94.5%. There was major improvement in the parameters for evaluating the program's quality, although they were still far from ideal. Public-awareness campaigns on the importance of neonatal screening are needed to increase the program's coverage. Setting postnatal day 3 as the standard Day for the Heel Stick Test would help ensure treatment at earlier ages, thus improving prognosis for affected infants. PMID- 19219253 TI - Characteristics and circumstances of falls leading to severe fractures in elderly people in Rio de Janeiro, Brazil. AB - The circumstances associated with falls among elderly people have been well described in some developed countries, but little is known about such accidents in middle-income countries. The aim of this paper is to report the circumstances and characteristics associated with falls leading to severe fractures among elderly people living in Rio de Janeiro, Brazil. Cases were obtained from two case-control studies on risk factors for severe fractures due to falls among individuals 60 years or older. Fallers were stratified according to clinical and socio-demographic variables and circumstances of the accident. Women comprised three-fourths of the sample. The femur was the most frequently fractured bone, more common among individuals over 70 years of age. Most falls occurred at home, between 6:00 AM and 6:00 PM, but the rooms where they occurred differed according to gender and age. Most individuals did not attribute their falls to tripping or slipping. This sample's data did not differ substantially from studies in developed countries. PMID- 19219254 TI - [High prevalence of hepatitis C in the district of Botafogo, municipality of Bebedouro, Sao Paulo State, Brazil, 2007]. AB - This article aimed to estimate the prevalence of hepatitis C in Botafogo, a district of Bebedouro, Sao Paulo State, Brazil, and investigate possible risk factors. One individual over 18 years of age was selected from each household to answer a questionnaire on socio-demographic variables and factors associated with hepatitis C. Blood samples were also drawn for immunoenzymatic tests. Positive HCV-antibody samples were submitted to viral RNA detection. HCV prevalence was 8.8% (95%CI: 5.8-11.7), and independent variables associated with risk of infection were: male gender, time of local residence > 30 years, and history of injected medication using non-disposable material, sterilized by boiling. The high prevalence of hepatitis C infection in this relatively isolated rural population appears to result from previous exposure to injections with inadequately sterilized material, with some evidence suggesting a specific elderly pharmacy employee who customarily applied such injections and may have been a chronic HCV carrier. PMID- 19219255 TI - Spine subdural hematoma: a rare complication associated with vitamin K antagonist (VKA). AB - Spinal subdural hematoma (SSDH) is a rare condition, which is difficult to diagnose, related to Vitamin K Antagonist. This a case report of a life threatening situation in a octogenarian patient with a history of recent atrial fibrillation that received K-Vitamin Antagonist (KVA) therapy. The history and the clinical assessment were normal at the admission, associated with increase in the coagulation parameters (INR >10). Twenty-four hours after the admission, the patient developed progressive tetraparesis and a Magnetic Resonance Imaging (MRI) was performed, disclosing the evidence of a SSDH (Figure 1). An emergency neurosurgical intervention was performed associated with normalization of the coagulation parameters. After the procedure, the patient presented improvement of the neurological symptoms. PMID- 19219256 TI - The Brazilian cardiology trainee: guidelines and freedom. PMID- 19219258 TI - Pharmacological stent deployment in the left anterior descending artery: late event indicators. AB - BACKGROUND: The efficacy of pharmacological stents in decreasing the incidence of cardiac events is not homogeneous for all lesions or patient subgroups. OBJECTIVE: 1) To evaluate the late clinical evolution of patients submitted to pharmacological stent implantation in atherosclerotic lesions of the left anterior descending artery; 2) to identify, among the clinical, angiographic and intravascular ultrasonographic characteristics, the ones predictive of cardiac event risk. METHODS: From May 2002 to August 2005, 205 patients were treated with 236 pharmacological stent implants, guided by the intravascular US (IVUS). RESULTS: After a mean follow-up period of 711 days, the rate of stent thrombosis was 0.48%, the same observed for acute myocardial infarction or revascularization surgery. The revascularization rate of the treated lesion was 7.31% and the general event rate was 10.24%. The event indicators, according to the multivariate analysis were the implant of more than one stent in the same artery, concentric lesions and the minimal intra-stent area measured by IVUS < 3.88 mm(2). CONCLUSION: Based on the data obtained, we conclude that the revascularization of the left anterior descending artery with pharmacological stent implant, chosen and optimized by IVUS, presents a low incidence of late events. The implant of two pharmacological stents for the treatment of long lesions was the main independent factor for the occurrence of late events. The final luminal area > 3.88 mm(2) obtained in the small reference-diameter segments is an independent indicator of event-free evolution. PMID- 19219259 TI - Validation of the Omron HEM 742 blood pressure monitoring device in adolescents. AB - BACKGROUND: Accurate blood pressure measurement is fundamental for scientific investigation or clinical decision-making. In this sense, it is important to verify the values provided by electronic devices. OBJECTIVE: To validate the Omron HEM 742 blood pressure monitoring device in adolescents according to criteria suggested by the British Hypertension Society. METHODS: A total of 150 adolescents aged between 10 and 16 years participated in the study. The automated Omron HEM 742 monitor was connected in Y to the mercury column auscultatory device, then three simultaneous measurements were taken, and the differences between the readings of the two devices were calculated. The intraclass correlation coefficient and Bland-Altman plot (agreement) were used to verify the relationship between both devices. Specificity and sensitivity of the device were determined by using the ROC curve. RESULTS: The comparison between the measurements showed an equal to or lower than 5mmHg difference in 67.3% of the systolic values, and 69.3% of the diastolic values; an equal to or lower than 10mmHg difference occurred in 87.3% and 90.6% of the systolic and diastolic values, respectively; an equal to or lower than 15mmHg difference was found in 96.6% of the systolic values and 97.3% of the diastolic values. These findings are consistent with a grade A according to the British Hypertension Society protocol. A marked consonance was observed between the values obtained by the automated monitor and this device was proven to be capable of identifying the presence or absence of high blood pressure. CONCLUSION: The Omron HEM 742 monitor was proved valid for blood pressure measurement in adolescents according to the criteria suggested by the British Hypertension Society. PMID- 19219260 TI - Prevalence of cardiovascular risk factors in a population of Brazilian industry workers. AB - BACKGROUND: Determining the cardiovascular risk factors is essential for the primary and secondary prevention of circulatory system diseases. OBJECTIVE: To obtain the prevalence of cardiovascular risk factors in a population of industry workers in Brazil. METHODS: Transversal cohort study, with a sociodemographic interview to identify cardiovascular risk factors, anthropometric and blood pressure measurements and capillary blood collection for blood glucose, cholesterol and triglyceride measurement in food industry workers of both sexes. RESULTS: A total of 1,047 workers were assessed, with 913 (87%) of them being males, with a mean age of 36 +/- 8 years. The frequency of a sedentary lifestyle was 83% and of overweight, 63%. Systemic arterial hypertension was identified in 28% of the individuals and 45% were in the pre-hypertension range. Alterations in the blood glucose levels were identified in 49% of the participants, as well as high levels of cholesterol and triglycerides in 7% and 11% of the population, respectively. The body mass index (BMI) levels were not associated to income, but there was an inverse association with the level of schooling. CONCLUSION: Overweight and a sedentary lifestyle are the main cardiovascular risk factors in a population of industry workers. PMID- 19219261 TI - Effect of frequency of physical exercise on glycemic control and body composition in type 2 diabetic patients. AB - BACKGROUND: Diabetes and cardiovascular disease have emerged as key threats to human health, and the risk is increased in individuals with visceral obesity. The consensus is that physical exercise should be part of the treatment of diabetes mellitus (DM). OBJECTIVE: To compare the influence of guided and structured physical exercise programs (SPEP), three to five times per week, during a period of 20 weeks, on glycemic control and body composition of type 2 diabetic patients (DM2). METHODS: The research was conducted at the Universidade Federal de Sao Paulo (Federal University School of Medicine in Sao Paulo). At the clinical visit, patients from the Control Group (CG) n=17, mean age 55.8 years, were encouraged to engage in a physical exercise program. Patients from Group 3x (G3), n=14, mean age 57.4 years, were to engage in 1 hour of physical exercise, 3x/week, and Group 5x (G5), n=9, mean age 58.8 years, followed the same protocol but 5x/week. Mean of 5 years since diagnosis in all groups. Classes consisted of a 5-minute warm-up, 30-minute treadmill walk at 70% of maximum heart rate, and 10 minute relaxation. BMI, abdominal circumference (AC), percentage of body fat (BF), capillary glycemia (CG), fasting glycemia (FG), and glycated hemoglobin (HbA1c) were assessed. RESULTS: A comparison was made between the baseline time point (B) and the 20th week (20th). BMI in G3 (B:29.5+/-2.9 vs. 20th: 28.3 +/- 2.2 Kg/sqm, p=0.005) and G5 (B:29.7+/-4.4 vs. 20th: 29.1 +/- 4.3 Kg/sqm, p=0.025); abdominal circumference in G5 (B:100.5+/-11.9 vs. 20th: 933 +/- 11.7 cm, p=0.001); BF in G3 (B:31+/-5.1 vs 20th: 26+/-5%, p=0.001) and G5 (B:32.4 +/- 5.4 vs. 20th: 30.3 +/- 6.9%, p=0.001); FG, G5 (B:150.8 +/- 47.5 vs. 20th: 109.2+/ 30.5 mg/dL, p=0.034), showed statistically significant differences. CG did not show statistically significant differences for these variables. CG showed a tendency to drop after physical exercise in G5. HbA1c showed no statistically significant differences in the three groups. CONCLUSION: G5 did better than G3 in most parameters assessed. However, results failed to show a decrease of HbA1c in DM2 patients. PMID- 19219262 TI - Physical exercise attenuates the cardiac autonomic deficit induced by nitric oxide synthesis blockade. AB - BACKGROUND: The nitric oxide (NO) synthesis blockade is characterized by an increase in the cardiac sympathetic activity and the physical training promotes the decrease in the sympathetic activity. OBJECTIVE: We investigated the effect of the NO synthesis blockade on the autonomic cardiovascular control in rats submitted to aerobic exercises during a 10-week period. METHODS: Male Wistar rats were divided in four groups: control rats, treated with chow food and water ad libitum for 10 weeks (CR); control rats, treated with N G-nitro-L-arginine methyl ester (L-NAME) during the last week (CRL); rats trained during 10 weeks on an electrical treadmill (TR); rats trained for 10 weeks and treated with L-NAME during the last week (TRL). The autonomic cardiovascular control was investigated in all groups with the use of a double blockade with methylatropine and propranolol and analysis of variability. RESULTS: The CRL and TRL groups presented hypertension. The CRL group presented tachycardia and predominance of the sympathetic tonus in heat rate (HR) measurement after the pharmacological autonomic blockade. The TR group presented bradycardia and lower intrinsic HR when compared to the others. The evaluation of the HR variability showed lower absolute and normalized values in the low frequency (LF) band in the CRL group. On the other hand, the TRL presented an increase in the LF band in absolute values. The analysis of variability of the systemic arterial pressure (SAP) showed that the CRL and TRL groups presented higher values in the LF band. CONCLUSION: The previous physical exercise prevented the deficit in the autonomic cardiac control induced by the treatment with L-NAME, but did not prevent the increase in the SAP variability. PMID- 19219263 TI - Prevalence of primary hyperaldosteronism in a systemic arterial hypertension league. AB - BACKGROUND: Until recently, primary hyperaldosteronism was considered a rare cause of secondary hypertension. However, in recent years, many studies have suggested that this disease can affect up to 20% of hypertensive individuals. OBJECTIVE: To determine the prevalence of primary hyperaldosteronism in hypertensive patients treated at the hypertension league of a university hospital. METHODS: Serum aldosterone and plasma renin activity levels were measured in 105 patients while they were undergoing standard antihypertensive treatment, with the exception of those using betablockers and spironolactone, in fasting condition and after rest in the supine position for 20 minutes. Those with an aldosterone/plasma renin activity ratio > 25 were submitted to the saline suppression test and, after the confirmation of the autonomy of aldosterone secretion, a computed tomography of the adrenals was performed. The results are presented as percentages and means and standard deviations. RESULTS: Of the 105 patients, 6.54% presented refractory hypertension. Nine presented an aldosterone/plasma renin activity ratio > 25 (8.5% of the total). Of these, 08 were submitted to the saline suppression test and 01 (with refractory hypertension) had the diagnosis of primary hyperaldosteronism confirmed (0.96% of the total). A computed tomography of the adrenals was performed, which showed normal results. CONCLUSION: The prevalence of primary hyperaldosteronism in the studied sample was 0.96% of the total. However, when only the patients with refractory hypertension were evaluated, the prevalence was 14.3%. PMID- 19219264 TI - Microneurography and venous occlusion plethysmography in heart failure: correlation with prognosis. AB - BACKGROUND: Microneurography and venous occlusion plethysmography can be considered methods of assessment of the sympathetic activity. OBJECTIVE: To evaluate the intensity of the sympathetic activity through microneurography and venous occlusion plethysmography in patients with heart failure (HF) and correlate this intensity with prognosis. METHODS: 52 patients with HF (ejection fraction < 45% at the echocardiogram): 12 with FCII and 40 with FCIV. After compensation, the muscular sympathetic nervous activity (MSNA) in the peroneal nerve (microneurography) and the muscular blood flow (MBF) in the forearm were evaluated (venous occlusion plethysmography). After an 18-month follow-up, the patients were divided in 3 groups: 12 with FCII, 19 with FCIV that did not die and 21 with FCIV that died. The intensity of the sympathetic activity was compared in the three different groups. RESULTS: Patients with FCII presented lower MSNA (p=0.026) and higher MBF (p=0.045) than the ones with FCIV that did not die. The patients with FCIV that died presented higher MSNA (p<0.001) and lower MBF (p=0.002) than the patients with FCIV that did not die. ROC curve: cutoff >53.5 impulses/min for MSNA (S=90.55. E=73.68%) and < 1.81 ml/min/100gr for MBF (S=90.4%. E=73.7%). Kaplan-Meier curve: higher survival with MSNA < 53.5 impulses/min (p<0.001), and/or MBF >1.81 ml/min/100gr (P<0.001). Logistic regression analysis: the higher the MSNA and the lower the MBF, the higher is the probability of death. CONCLUSION: The intensity of the MSNA and the MBF can be considered prognostic markers in advanced HF. PMID- 19219265 TI - Correlation of myocardial interstitial collagen in the right ventricular septum with ventricular function of patients with ischemic cardiomyopathy. AB - BACKGROUND: Myocardial collagen content influences ventricular relaxation, contraction, and morphology. Its relationship with ventricular function in patients (Pts) with ischemic cardiomyopathy (ICMP) has not yet been fully studied in humans. OBJECTIVE: To assess the relationship between interstitial collagen content in non-infarcted areas of the right ventricular septum and ventricular function in ICMP. METHODS: 31 pts with coronary artery disease were divided into four groups as follows:The control group consisted of 7 pts with normal left (LVEF) and right (RVEF) ventricular ejection fraction (group C); Group 1: 5 patients with RVEF < 40%; Group 2: 9 pts with LVEF < 40%; and Group 3, 10 pts with biventricular dysfunction. RVEF and LVEF were measured by radionuclide angiography. For quantitative analysis of interstitial collagen volume fraction (CVF), endomyocardial biopsy specimens were taken from the right ventricle and stained with picrosirius red. RESULTS: Mean CVF was significantly higher in group 3, compared with the control group and with groups 1 and 2 (30.2 +/- 7.9% vs. 6.8 +/- 3.3% vs. 15.8 +/- 4.1% vs. 17.5+/-7.7%, respectively; p =0.0001). It was also significantly higher in patients belonging to group 2, compared with those in the control group (17.5 +/- 7.7% vs. 6.8 +/- 3.3%, p =0.0001). CVF was inversely correlated with RVEF (r = - 0.50, p = 0.003) and LVEF (r = -0.70, p = 0.0001). CONCLUSION: In ICMP, CVF is elevated in non-infarcted areas of the right ventricular septum and inversely correlated with right and left ventricular function. PMID- 19219266 TI - Myocardial fibrosis and ventricular remodeling in severe chronic aortic regurgitation. AB - BACKGROUND: Significant symptomatic chronic aortic regurgitation (AR) leads to considerable left ventricular remodeling at the expense of myocyte hypertrophy and extracellular matrix remodeling. The relevance of interstitial fibrosis concentration in these patients is unknown. We analyzed the degree of fibrosis in the left ventricle (LV) in symptomatic patients with AR submitted to surgical treatment, and its relationship with functional and anatomical characteristics. OBJECTIVE: To evaluate myocardial fibrosis in chronic severe aortic regurgitation. METHODS: Twenty-eight patients with chronic symptomatic AR (16 with normal LV function and 12 with LV dysfunction) were selected and assessed pre- and postoperatively by echocardiography. Functional capacity was measured using maximal oxygen consumption (VO2max) through the cardiopulmonary test. Myocardial fibrosis volume fraction (MFV) was quantified through endomyocardial biopsy performed in all patients during surgery. We compared the histopathologic results with a nine-patient control group. RESULTS: The mean age was 39 +/- 12 years, 75% of the patients were male, and the rheumatic etiology accounted for 84% of the cases. Twenty-five patients remained in FC l and ll at the end of the study, and there was a significant reduction of the LV diameters between the preoperative and late postoperative timepoints. Three deaths occurred but they were not related to postoperative ventricular dysfunction. The parameters of the cardiopulmonary test were similar between pre- and postoperative timepoints. MFV in patients with AR was significantly higher than in the control group (3.47 +/- 1.9% vs 0.82 +/- 0.96%, respectively, p=0.001). There was no statistical correlation among LV fibrosis and LV diameters, LVEF and MVO2. CONCLUSION: In patients with significant symptomatic AR, the presence of limited myocardial fibrosis was not associated with clinical, echocardiographic or functional complications. PMID- 19219267 TI - Glycoprotein IIb/IIIa inhibitors in clinical practice. PMID- 19219268 TI - Case 1/2009: 21-month-old infant with partial atrioventricular septal defect, consisting of atrial septal defect and tricuspid insufficiency. PMID- 19219270 TI - Quinine levels in patients with uncomplicated falciparum malaria in the Amazon region of Brazil. AB - We examined the plasmatic concentrations of quinine in patients with uncomplicated falciparum malaria in an endemic area of the Amazon region in Brazil in a prospective clinical trial, in which a standard three-day course of oral quinine plus doxycycline was used. We measured the quinine in the plasma samples on days 0 and 3 by high performance liquid chromatography. The mean concentration of quinine was 6.04 +/-2.21 microg/mL in male patients and 5.98 +/ 1.95 microg/mL in female patients. No significant differences in quinine concentration were observed between these two groups. All samples collected before starting treatment were negative for quinine. This information could help in the development of strategies for the rational use of antimalarial drugs in Brazil. PMID- 19219271 TI - Lamivudine for chronic hepatitis B: a brief review. AB - Until recently, the only generally approved treatment for chronic hepatitis B was alpha-interferon; however, it gives only moderate efficacy in terms of sustained response (biochemical, virological and histological). In fact, only 20% to 40% of treated patients respond to therapy, with lower percentages (~ 10%) among patients infected with precore-mutant strains of HBV (HBeAb HBV-DNA positive). The FDA of the USA approved the use of lamivudine in adult patients affected by chronic hepatitis B in 1998. In this review, we focused on the pharmacokinetic and pharmacodynamic properties and efficacy and tolerability of lamivudine in the treatment of chronic hepatitis B cases that are both HBeAg and anti-HBe-positive. PMID- 19219272 TI - Hepatitis C virus detection in the semen of infected patients. AB - Though HCV infection is a serious public health problem, some aspects of its biology are still not well understood, such as its transmission through seminal fluid and sexual transmission. We looked for HCV in the semen of infected patients. Thirteen patients were included. Semen fractions (seminal plasma, leukocytes and spermatozoa) were separated with 45% and 90% Percoll gradients. The HCV-RNA in blood and semen fractions was extracted using the same protocol (AMPLICOR Roche) and was detected using the qualitative Roche Amplicor test and by agarose gel electrophoresis, with ethidium bromide staining. The mean age of the patients was 40.7 years. Risk factors for the acquisition of HCV included injectable and inhaled drug use in six (42.8%), blood transfusion in four (28.6%), and no risk factors in four (28.6%) patients. Genotype 1 was detected in 62% of the patients, followed by genotype 3 in 23% and genotype 2 in 15%. All blood samples were positive, regardless of the technique used for detection. All semen samples identified by Roche Amplicor and analyzed by agarose gel electrophoresis were negative. Among the 52 semen samples (total and fractions) identified by the Roche Amplicor method, 45 (87%) were inhibited. A negative result was recorded for one (1.9%) total semen sample, one (1.9%) leukocyte and four (7.7%) seminal plasma fractions. Only one (1.9%) sample of the spermatozoon fraction was positive. The results obtained suggested false-negative reactions for the semen samples. PMID- 19219273 TI - Histological response study of chronic viral hepatitis C patients treated with interferon alone or combined with ribavirin. AB - Chronic hepatitis C is often a progressive, fibrotic disease that can lead to cirrhosis and other complications. The recommended therapy is a combination of interferon and ribavirin. Besides its antiviral action, interferon is considered to have antifibrotic activity. We examined the outcome of hepatic fibrosis and inflammation in chronic hepatitis C patients who were non-responders to interferon. We made a case series, retrospective study, based on revision of medical records and reassessment of liver biopsies. For inclusion, patients should have been treated with interferon alone or combined with ribavirin, with no virological response (non responders and relapsers) and had a liver biopsy before and after treatment. Histological evaluation included: i-outcome of fibrosis and necroinflammation; ii-annual fibrosis progression rate evaluation, before and after treatment. Seventy-five patients were included. Fifty-seven patients (76%) did not show progression of fibrosis after treatment, compared to six (8%) before treatment (p < 0.001). The mean annual fibrosis progression rate was significantly reduced after treatment (p = 0.036). Inflammatory activity improved in 19 patients (25.3%). The results support the hypothesis of an antifibrotic effect of interferon-based therapy, in non-responder patients. There was evidence of anti-inflammatory effects of treatment in some patients. PMID- 19219274 TI - Cost-effectiveness of entecavir versus lamivudine for the suppression of viral replication in chronic hepatitis B patients in Brazil. AB - Hepatitis B virus infection is an important public-health issue. Chronic patients have a higher risk of death due to complications, which increases health-care expenses in. Cost-effectiveness analysis of entecavir (ETV) versus lamivudine (LVD) for treatment of chronic hepatitis B, in e antigen (AgHBe)-positive and negative patients, based on two phase 3, controlled and randomized studies. A decision analysis model was developed, using the following endpoints: cost per patient with undetectable viral load and cost per quality life year (QALY) gained. Risks for complications (compensated or decompensated cirrhosis and hepatocellular carcinoma) were based on the cohort study REVEAL, published in 2006. The REVEAL parameters were applied to the results of the viral load levels obtained from the clinical assay data. The complication costs were based on a study of the disease cost conducted in Brazil, in 2005. The cost data were obtained predominantly from Sistema Unico de Saude [SUS - Brazilian public health system] payment tables and drug price lists. The utility data were obtained from literature and life expectancy information was based on IBGE data. The analysis perspective was that of SUS. A discount rate of 3% per year was used. For the horizon of time of 10 years, the ETV had an incremental cost of approximately two million Brazilian Reais (R$) compared to LVD. Reducing the number of complications, ETV treatment reduced costs by around 3 million, reducing final costs by 1 million, for AgHBe-positive patients. ETV also reduced the incremental cost per QALY gained. ETV was found to be the most cost-effective alternative for AgHBe-positive and negative patients. PMID- 19219275 TI - Genetic characterization and evolutionary inference of TNF-alpha through computational analysis. AB - TNF-alpha is an important human cytokine that imparts dualism in malaria pathogenicity. At high dosages, TNF-alpha is believed to provoke pathogenicity in cerebral malaria; while at lower dosages TNF-alpha is protective against severe human malaria. In order to understand the human TNF-alpha gene and to ascertain evolutionary aspects of its dualistic nature for malaria pathogenicity, we characterized this gene in detail in six different mammalian taxa. The avian taxon, Gallus gallus was included in our study, as TNF-alpha is not present in birds; therefore, a tandemly placed duplicate of TNF-alpha (LT-alpha or TNF-beta) was included. A comparative study was made of nucleotide length variations, intron and exon sizes and number variations, differential compositions of coding to non-coding bases, etc., to look for similarities/dissimilarities in the TNF alpha gene across all seven taxa. A phylogenetic analysis revealed the pattern found in other genes, as humans, chimpanzees and rhesus monkeys were placed in a single clade, and rats and mice in another; the chicken was in a clearly separate branch. We further focused on these three taxa and aligned the amino acid sequences; there were small differences between humans and chimpanzees; both were more different from the rhesus monkey. Further, comparison of coding and non coding nucleotide length variations and coding to non-coding nucleotide ratio between TNF-alpha and TNF-beta among these three mammalian taxa provided a first hand indication of the role of the TNF-alpha gene, but not of TNF-beta in the dualistic nature of TNF-alpha in malaria pathogenicity. PMID- 19219276 TI - Prevalence of resistance-associated mutations in human immunodeficiency virus type 1-positive individuals failing HAART in Rio de Janeiro, Brazil. AB - We investigated the occurrence of HIV-1 antiretroviral resistance in individuals failing to respond to highly active antiretroviral therapy (HAART) attended by RENAGENO from 2001-2004. One hundred and seventeen patients were selected for this study; their plasma viral RNA was extracted and the PR and RT genes sequenced to examine subtype, genetic polymorphisms and mutations associated with resistance to antiretroviral drugs. HIV-1 sequence analysis showed that 86/100 (86%) were infected with subtype B, 7/100 (7%) with subtype F and 7/100 (7%) with RT/PR hybrid forms (2 D/B, 2 F/B, 2 B/F and 1 D/F). In 14 (12%) of the samples, the subtype was not determined. The prevalence of resistance mutations was high (93.1%), mainly in the RT gene. The most prevalent resistance mutations were: M184V (60.7%), T215Y (49.6%) and M41L (46.7%) in the RT gene and L90M (19.6%), M46I (16.2%) and D30N (12.8%) in the PR gene. The frequency of resistance mutations tended to increase from the first to the second therapeutic scheme failure (p=0.079); but it stabilized after subsequent failures (p=0.875). Our finding of a high frequency of drug resistant HIV-1 samples supports the need for continuous genotypic monitoring of patients failing HAART. PMID- 19219277 TI - Invasive aspergillosis in hematopoietic stem cell transplant recipients: a retrospective analysis. AB - Invasive aspergillosis (IA) currently is an important cause of mortality in subjects undergoing hematopoietic stem cell transplants (HSCT) and is also an important cause of opportunistic respiratory and disseminated infections in other types of immunocompromised patients. We examined the medical records of 24 cases of proven and probable invasive aspergillosis (IA) at the Hospital de Clinicas of the Federal University of Parana, Brazil, from January 1996 to October 2006. During this period occurred a mean of 2.2 cases per year or 3.0 cases per 100 HSTC transplants. There was a significant relationship between structural changes in the bone marrow transplant (BMT) Unit and the occurrence of IA cases (p=0.034, relative risk (RR) = 2.47). Approximately 83% of the patients died due to invasive fungal infection within 60 days of follow up. Some factors tended to be associated with mortality, but these associations were not significant. These included corticosteroid use, neutropenia (<100 cells/mm(3)) at diagnosis, patients that needed to change antifungal therapy because of toxicity of the initial first-line regimen and disseminated disease. These factors should be monitored in BMT units to help prevent IA. Physicians should be aware of the risk factors for developing invasive fungal infections and try to reduce or eliminate them. However, once this invasive disease begins, appropriate diagnostic and treatment measures must be implemented as soon as possible in order to prevent the high mortality rates associated with this condition. PMID- 19219278 TI - An outbreak of Candida spp. bloodstream infection in a tertiary care center in Bogota, Colombia. AB - Several cases of Candida bloodstream infections were documented from June to October 2004 at a tertiary care center in Bogota, Colombia. Since no cases of candidemia had occurred during the preceding four months, an outbreak was declared. As a result, a microbiological study, a revision of infection control practices and a case-control study were performed. In all, 18 cases of candidemia were ascertained. Parenteral nutrition (p=0.04), presence of a central line (p=0.03), and severity of illness (p=0.03) were associated with candidemia in bivariate analysis. Diverse Candida species were observed. Candida parapsilosis contamination was found in plastic containers used for transient intravenous (IV) medication storage at the bedside, plastic bags reused for the transportation of IV medicines and cotton used for disinfection of IV ports. Poor infection control practices were widely observed. The outbreak was controlled after elimination of plastic bags used for transportation, instauration of daily disinfection of IV medication containers, acquisition of sterile alcohol swabs for port disinfection and staff education. It was concluded that candidemia was associated with previously-described risk factors and that poor infection-control practices were likely responsible for the outbreak. PMID- 19219279 TI - Molecular investigation of a fungemia outbreak due to Candida parapsilosis in an intensive care unit. AB - We investigated a nosocomial cluster of four Candida parapsilosis fungemia episodes that occurred in a neurological intensive care unit over a two-week period. The four infected patients had received parenteral nutrition through central lines, and all four had catheter-related candidemia. All of the isolates were susceptible to all of the antifungals tested, including amphotericin B, fluconazole, voriconazole, and caspofungin. They had strictly related fingerprints, based on randomly amplified polymorphic DNA analysis. Additional DNA sequencing data revealed that they were same strain. Although no isolate of Candida parapsilosis was recovered from other clinical, surveillance, or environmental samples, nosocomial spread of this yeast ceased, following the reinforcement of infection-control measures. Candida parapsilosis may require an intravascular foreign body to cause fungemia, but this outbreak shows that it can be transmitted nosocomially and can cause epidemics. PMID- 19219280 TI - Treatment of invasive fungal infections: stability of voriconazole infusion solutions in PVC bags. AB - Voriconazole is a novel broad-spectrum antifungal drug, employed in the treatment of invasive fungal infections, and represents an alternative to amphotericin B treatment. The manufacturer recommends that any unused reconstituted product should be stored at 2 masculineC to 8 masculineC, for no more than 24 h, but no recommendations about i.v. infusion solutions are given. Previous works have reported on the stability of voriconazole in polyolefin bags and just one in 5% dextrose polyvinyl chloride (PVC) bags, at a 4 mg.mL-1 concentration. In this work, the stability of voriconazole as an i.v. infusion solution in 0.9% sodium chloride and in 5% dextrose, in PVC bags, at 0.5 mg.mL-1, stored at 4 masculineC and at room temperature, protected from light, was evaluated. These infusion solutions were analyzed for a 21-day period. Chemical stability was evaluated by HPLC assay. Visual inspection was performed and pH of the solutions was measured. No color change or precipitation in the solutions was observed. The drug content remained above 90% for 11 days in 0.9% sodium chloride and for 9 days in 5% dextrose solutions. The i.v. infusion solutions stored at room temperature were not stable. At room temperature, the voriconazole content dropped down to 88.3 and 86.6%, in 0.9% sodium chloride or 5% dextrose solutions, respectively, two days after admixture. Assays performed at the end of the study suggest the sorption of voriconazole by the PVC bags. The results of this study allow cost effective batch production in the hospital pharmacy. PMID- 19219281 TI - Rates of antimicrobial resistance in Latin America (2004-2007) and in vitro activity of the glycylcycline tigecycline and of other antibiotics. AB - As a part of the Tigecycline Evaluation and Surveillance Trial (T.E.S.T.), Gram positive and Gram-negative bacterial isolates were collected from 33 centers in Latin America (centers in Argentina, Brazil, Chile, Colombia, Guatemala, Honduras, Jamaica, Mexico, Panama, Puerto Rico, and Venezuela) from January 2004 to September 2007. Argentina and Mexico were the greatest contributors of isolates to this study. Susceptibilities were determined according to Clinical Laboratory Standards Institute guidelines. Resistance levels were high for most key organisms across Latin America: 48.3% of Staphylococcus aureus isolates were methicillin-resistant while 21.4% of Acinetobacter spp. isolates were imipenem resistant. Extended-spectrum beta-lactamase were reported in 36.7% of Klebsiella pneumoniae and 20.8% of E. coli isolates. Tigecycline was the most active agent against Gram-positive isolates. Tigecycline was also highly active against all Gram-negative organisms, with the exception of Pseuodomonas aeruginosa, against which piperacillin-tazobactam was the most active agent tested (79.3% of isolates susceptible). The in vitro activity of tigecycline against both Gram-positive and Gram-negative isolates indicates that it may be an useful tool for the treatment of nosocomial infections, even those caused by organisms that are resistant to other antibacterial agents. PMID- 19219282 TI - Inadequate timing between corticosteroid and antibiotics applications increases mortality due to sepsis. AB - This study tested the hypothesis that the use of corticosteroids prior to antibiotics can lower the mortality rate in severe infections by S. aureus or Gram-negative bacilli, using an animal model. This study was a prospective and controlled study, placed in a university laboratory. Seven hundred and sixty mice distributed into three groups (Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae infected). The interventions in each group were: I) infection control (intra-peritoneal); II) treatment solely with antibiotics (teicoplanin or amikacin); III) antibiotics administered prior to the corticosteroid (methylprednisolone); IV) antibiotics administered after the corticosteroid. Mortality in the E. coli group, subgroup I: 100%; subgroup II: 55% (p<0.001); subgroup III: 62.5% (p=0.2488, compared to subgroup II); subgroup IV: 20% (p<0.01 compared to subgroups II and III). Mortality in the K. pneumoniae group: subgroup I: 100%; subgroup II: 72.5% (p<0.01); subgroup III: 80% (p=0.215 compared to subgroup II); subgroup IV: 45% (p<0.01 compared to subgroups II and III). Mortality in the S. aureus group: subgroup I: 82.5%; II: 42.5% (p<0.001); subgroup III: 77.5% (p=0.2877 compared to subgroup I); subgroup IV: 32.5% (p=0.1792 compared to subgroup II). The use of corticosteroids prior to antibiotics lowered the mortality rate caused by Gram-negative bacteria and did not affect the mortality caused by S. aureus. When used after starting treatment with antibiotics, the corticosteroid was not superior to the use of antibiotics alone in the case of the Gram-negative bacteria, and was not significantly different from non-treatment of the infection, in the case of S. aureus. PMID- 19219283 TI - Comparison of PCR-based molecular markers for the characterization of Proteus mirabilis clinical isolates. AB - Proteus mirabilis is one of the most important pathogens associated with complicated urinary tract infections (acute pyelonephritis, bladder infections, kidney stones) and bacteremia, affecting patients with anatomical abnormalities, immunodeficiency, and long-term urinary catheterization. For epidemiological purposes, various molecular typing methods, such as pulse-field gel electrophoresis (PFGE) or ribotyping, have been developed for this pathogen. However, these methods are labor intensive and time-consuming. We evaluated the discriminatory power of several PCR-based fingerprinting methods (RAPD, ISSR, ERIC-PCR, BOX-PCR and rep-PCR) for P. mirabilis clinical isolates. Typing patterns and clustering analysis indicated that RAPD, BOX-PCR and ERIC-PCR differentiated P. mirabilis strains from Escherichia coli, Hafnia alvei, and Morganella morganii. With the exception of rep-PCR, the methods gave medium to high discriminatory efficiency in P. mirabilis. In general, the results obtained with RAPD, BOX-PCR and ERIC-PCR were in good agreement. We concluded that a combination of ERIC-PCR and BOX-PCR results is a rapid and reliable alternative for discrimination among P. mirabilis clinical isolates, contributing to epidemiological studies. PMID- 19219284 TI - Ribotyping, biotyping and capsular typing of Haemophilus influenzae strains isolated from patients in Campinas, southeast Brazil. AB - Forty-five Haemophilus influenzae strains isolated from patients were characterized based on biochemical characteristics. Their capsular types were determined by polymerase chain reaction (PCR); they were compared, using two molecular methods [ribotyping with a specific DNA probe amplified from the 16S rDNA region from H. influenzae and through restriction fragment length polymorphism (RLFP) of an amplified 16S DNA region]. The strains were better discriminated by the ribotyping technique that used the 16S probe and by the combination of both techniques. Biotypes I and IV were the most common, followed by biotypes VI, VIII and III. Biotypes II and VII were not found. Most of the capsular samples were nontypable (89%), with capsular types a and b found in 2 and 9% of the samples, respectively. We concluded that there is a very close genetic identity among pathogenic and non-pathogenic strains. PMID- 19219285 TI - Postnatal acquired toxoplasmosis patients in an infectious diseases reference center. AB - Infection caused by Toxoplasma gondii, toxoplasmosis, is one of the most frequent zoonoses in the world; it normally affects both genders equally. Humans are one of several possible intermediate hosts, and the disease is oligosymptomatic in most cases. Vertical transmission is an important cause of fetal malformation and sequels in newborns. Approximately 10% of postnatal cases present multiple manifestations, ranging from low fever and mild lymphadenopathy to severe encephalitis. In moderate cases, lesions such as retinochoroiditis may emerge during acute infection or even years later. We analyzed 313 cases of toxoplasmosis from 1992 to 2004, including 261 acute cases. Most patients were women (68.1%), and 39% of these were pregnant. Among acute infection cases, 64.8% presented symptomatic disease; the most frequent manifestations were lymphadenomegaly (59.8%), fever (27.2%), headache (10.7%), asthenia (10%), weight loss (8.4%), myalgia (8%), retinochoroiditis (3.4%) and hepatosplenomegaly (1.5%). Although ocular lesions by T. gondii are well documented as a possible consequence of postnatal infection, two patients developed retinochoroiditis only two years after primary infection. This demonstrates the need for toxoplasmosis case surveillance, even long after acute manifestations. PMID- 19219286 TI - Tatumella ptyseos causing severe human infection: report of the first two Brazilian cases. AB - Tatumella ptyseos is the type species of the Tatumella genus (Enterobacteriaceae). This fermentative Gram-negative rod has only rarely been reported as a cause of human infections; there is very little information about it in the medical literature. We report here the first two Brazilian cases of T. ptyseos infections, both evolving to severe sepsis. PMID- 19219287 TI - Ventilator-associated pneumonia with Col-S strains: a successful comeback of colistin! AB - Emergence of multi and pan-drug resistant Gram-negative bacteria causing nosocomial infections in intensive care settings has become a challenge for clinicians. The mortality rate of ventilator-associated pneumonia (VAP) is known to increase when the initial microbiological diagnosis and antimicrobial therapy are inappropriate. We present a case of a 18-year-old man, who after being admitted following an accident, had developed VAP due to multi-drug resistant Pseudomonas aeruginosa and Acinetobacter spp. and had a downhill clinical course despite broad-spectrum antibiotic treatment. The strains were found to be Col-S, as the susceptibility was tested. Colistin was instituted, with remarkable recovery. It is imperative to diagnose VAP with multi-drug resistant strains as early as possible; colistin, the 'last resort' antibiotic, if instituted with proper monitoring at the right time, can be life saving. PMID- 19219288 TI - Extensively drug-resistant tuberculosis: a case report and literature review. AB - Extensively drug-resistant tuberculosis (XDR-TB) is an emerging health problem that threatens tuberculosis (TB) control worldwide, since suitable treatment for this disease has not yet been found. We report a case of secondary pulmonary XDR TB in a 54-year-old, HIV-negative male from Goiania, Brazil. The patient had long standing pulmonary tuberculosis (nine years) with extensive bilateral lung damage and had been treated with multiple antituberculosis drugs (self-administered) before XDR-TB diagnosis. The strain of Mycobacterium tuberculosis was resistant to R- rifampicin, H-isoniazid, E-ethambutol, Eto-ethionamide, Ofx-ofloxacin, and Am-amikacin. This patient died with multiple organ failure due to sepsis secondary to bacterial pneumonia. PMID- 19219289 TI - Primary gastric fundus tuberculosis in immunocompetent patient: a case report and literature review. AB - We report on a 29-year-old Pakistani man who presented to the clinic with epigastric pain, of one-month duration. He did not report fever, cough, vomiting blood, passing black stools, loss of appetite or diarrhea. However, he had lost 7 kg since his symptoms had begun. Clinical examination was unremarkable. Laboratory results were within normal limits. An abdominal CT scan showed a mass with enhancement in the stomach. Gastric endoscopy revealed an ulcerative mass in the fundus. An endoscopic-biopsy specimen revealed caseating granulomas with acid fast bacilli. The patient was diagnosed to have primary gastric tuberculosis, and antituberculous medications were initiated. Cultures of the gastric mass subsequently grew Mycobacterium tuberculosis sensitive to isoniazid and rifampcin. Follow-up after six months showed a good response to treatment; an upper gastrointestinal tract endoscopy after six months was normal. PMID- 19219290 TI - Acute hepatitis due to dengue virus in a chronic hepatitis patient. AB - We present a case of acute hepatitis caused by dengue virus, with a significant increase in aspartate transferase and alanine transferase levels in a chronic hepatitis patient attended at the Cane Sugar Planters Hospital of Campos dos Goytacazes, RJ. PMID- 19219291 TI - Leprosy with necrosis in granulomatous reaction. PMID- 19219292 TI - IV Miguel R. Covian Symposium: new challenges for physiologists. PMID- 19219293 TI - Ca2+ signaling in pancreatic acinar cells: physiology and pathophysiology. AB - The pancreatic acinar cell is a classical model for studies of secretion and signal transduction mechanisms. Because of the extensive endoplasmic reticulum and the large granular compartment, it has been possible--by direct measurements- to obtain considerable insights into intracellular Ca2+ handling under both normal and pathological conditions. Recent studies have also revealed important characteristics of stimulus-secretion coupling mechanisms in isolated human pancreatic acinar cells. The acinar cells are potentially dangerous because of the high intra-granular concentration of proteases, which become inappropriately activated in the human disease acute pancreatitis. This disease is due to toxic Ca2+ signals generated by excessive liberation of Ca2+ from both the endoplasmic reticulum and the secretory granules. PMID- 19219294 TI - The inhibitory role of sympathetic nervous system in the Ca2+-dependent proteolysis of skeletal muscle. AB - Mammalian cells contain several proteolytic systems to carry out the degradative processes and complex regulatory mechanisms to prevent excessive protein breakdown. Among these systems, the Ca2+-activated proteolytic system involves the cysteine proteases denoted calpains, and their inhibitor, calpastatin. Despite the rapid progress in molecular research on calpains and calpastatin, the physiological role and regulatory mechanisms of these proteins remain obscure. Interest in the adrenergic effect on Ca2+-dependent proteolysis has been stimulated by the finding that the administration of beta2-agonists induces muscle hypertrophy and prevents the loss of muscle mass in a variety of pathologic conditions in which calpains are activated. This review summarizes evidence indicating that the sympathetic nervous system produces anabolic, protein-sparing effects on skeletal muscle protein metabolism. Studies are reviewed, which indicate that epinephrine secreted by the adrenal medulla and norepinephrine released from adrenergic terminals have inhibitory effects on Ca2+ dependent protein degradation, mainly in oxidative muscles, by increasing calpastatin levels. Evidence is also presented that this antiproteolytic effect, which occurs under both basal conditions and in stress situations, seems to be mediated by beta2- and beta3-adrenoceptors and cAMP-dependent pathways. The understanding of the precise mechanisms by which catecholamines promote muscle anabolic effects may have therapeutic value for the treatment of muscle-wasting conditions and may enhance muscle growth in farm species for economic and nutritional purposes. PMID- 19219295 TI - Roles of mitogen-activated protein kinases and angiotensin II in renal development. AB - Experimental and clinical evidence suggests that angiotensin II (AII) participates in renal development. Renal AII content is several-fold higher in newborn rats and mice than in adult animals. AII receptors are also expressed in higher amounts in the kidneys of newborn rats. The kidneys of fetuses whose mother received a type 1 AII receptor (AT1) antagonist during gestation present several morphological alterations. Mutations in genes that encode components of the renin-angiotensin system are associated with autosomal recessive renal tubular dysgenesis. Morphological changes were detected in the kidneys of 3-week old angiotensin-deficient mice. Mitogen-activated protein kinases (MAPKs) are important mediators that transduce extracellular stimuli to intracellular responses. The MAPK family comprises three major subgroups, namely extracellular signal-regulated protein kinase (ERK), c-jun N-terminal kinases (JNK), and p38 MAPK (p38). Important events in renal growth during nephrogenesis such as cellular proliferation and differentiation accompanied by apoptosis on a large scale can be mediated by MAPK pathways. A decrease in glomerulus number was observed in embryos cultured for 48 and 120 h with ERK or p38 inhibitors. Many effects of AII are mediated by MAPK pathways. Treatment with losartan during lactation provoked changes in renal function and structure associated with alterations in AT1 and type 2 AII (AT2) receptors and p-JNK and p-p38 expression in the kidney. Several studies have shown that AII and MAPKs play an important role in renal development. However, the relationship between the effects of AII and MAPK activation on renal development is still unclear. PMID- 19219296 TI - Focal adhesion kinase signaling in cardiac hypertrophy and failure. AB - Focal adhesion kinase (FAK) is a broadly expressed tyrosine kinase implicated in cellular functions such as migration, growth and survival. Emerging data support a role for FAK in cardiac development, reactive hypertrophy and failure. Data reviewed here indicate that FAK plays a critical role at the cellular level in the responses of cardiomyocytes and cardiac fibroblasts to biomechanical stress and to hypertrophic agonists such as angiotensin II and endothelin. The signaling mechanisms regulated by FAK are discussed to provide insight into its role in the pathophysiology of cardiac hypertrophy and failure. PMID- 19219297 TI - Acute and chronic electrical activation of baroreceptor afferents in awake and anesthetized subjects. AB - Electrical stimulation of baroreceptor afferents was used in the 1960's in several species, including human beings, for the treatment of refractory hypertension. This approach bypasses the site of baroreceptor mechanosensory transduction. Chronic electrical stimulation of arterial baroreceptors, particularly of the carotid sinus nerve (Hering's nerve), was proposed as an ultimate effort to treat refractory hypertension and angina pectoris due to the limited nature of pharmacological therapy available at that time. Nevertheless, this approach was abandoned in the early 1970's due to technical limitations of implantable devices and to the development of better-tolerated antihypertensive medications. More recently, our laboratory developed the technique of electrical stimulation of the aortic depressor nerve in conscious rats, enabling access to hemodynamic responses without the undesirable effect of anesthesia. In addition, electrical stimulation of the aortic depressor nerve allows assessment of the hemodynamic responses and the sympathovagal balance of the heart in hypertensive rats, which exhibit a well-known decrease in baroreflex sensitivity, usually attributed to baroreceptor ending dysfunction. Recently, there has been renewed interest in using electrical stimulation of the carotid sinus, but not the carotid sinus nerve, to lower blood pressure in conscious hypertensive dogs as well as in hypertensive patients. Notably, previous undesirable technical outcomes associated with electrical stimulation of the carotid sinus nerve observed in the 1960's and 1970's have been overcome. Furthermore, promising data have been recently reported from clinical trials that evaluated the efficacy of carotid sinus stimulation in hypertensive patients with drug resistant hypertension. PMID- 19219298 TI - Central actions of glucocorticoids in the control of body fluid homeostasis: review. AB - The involvement of the hypothalamic-pituitary-adrenal axis in the control of body fluid homeostasis has been extensively investigated in the past few years. In the present study, we reviewed the recent results obtained using different approaches to investigate the effects of glucocorticoids on the mechanisms of oxytocin and vasopressin synthesis and secretion in response to acute and chronic plasma volume and osmolality changes. The data presented here suggest that glucocorticoids are not only involved in the mechanisms underlying the fast release but also in the transcriptional events that lead to decreased synthesis and secretion of these neuropeptides, particularly oxytocin, under diverse experimental conditions of altered fluid volume and tonicity. The endocannabinoid system, through its effects on glutamatergic neurotransmission within the hypothalamus and the nuclear factor kappaB-mediated transcriptional activity, seems to be also involved in the specific mechanisms by which glucocorticoids exert their central effects on neurohypophyseal hormone synthesis and secretion. PMID- 19219299 TI - Unconventional neurotransmitters, neurodegeneration and neuroprotection. AB - Neurotransmitters are also involved in functions other than conventional signal transfer between nerve cells, such as development, plasticity, neurodegeneration, and neuroprotection. For example, there is a considerable amount of data indicating developmental roles for the glutamatergic, cholinergic, dopaminergic, GABA-ergic, and ATP/adenosine systems. In this review, we discuss the existing literature on these 'new' functions of neurotransmitters in relation to some unconventional neurotransmitters, such as the endocannabinoids and nitric oxide. Data indicating both transcriptional and post-transcriptional modulation of endocannabinoid and nitrinergic systems after neural lesions are discussed in relation to the non-conventional roles of these neurotransmitters. Knowledge of the roles of neurotransmitters in brain functions other than information transfer is critical for a more complete understanding of the functional organization of the brain and to provide more opportunities for the development of therapeutical tools aimed at minimizing neuronal death. PMID- 19219300 TI - Learning about brain physiology and complexity from the study of the epilepsies. AB - The brain is a complex system, which produces emergent properties such as those associated with activity-dependent plasticity in processes of learning and memory. Therefore, understanding the integrated structures and functions of the brain is well beyond the scope of either superficial or extremely reductionistic approaches. Although a combination of zoom-in and zoom-out strategies is desirable when the brain is studied, constructing the appropriate interfaces to connect all levels of analysis is one of the most difficult challenges of contemporary neuroscience. Is it possible to build appropriate models of brain function and dysfunctions with computational tools? Among the best-known brain dysfunctions, epilepsies are neurological syndromes that reach a variety of networks, from widespread anatomical brain circuits to local molecular environments. One logical question would be: are those complex brain networks always producing maladaptive emergent properties compatible with epileptogenic substrates? The present review will deal with this question and will try to answer it by illustrating several points from the literature and from our laboratory data, with examples at the behavioral, electrophysiological, cellular and molecular levels. We conclude that, because the brain is a complex system compatible with the production of emergent properties, including plasticity, its functions should be approached using an integrated view. Concepts such as brain networks, graphics theory, neuroinformatics, and e-neuroscience are discussed as new transdisciplinary approaches dealing with the continuous growth of information about brain physiology and its dysfunctions. The epilepsies are discussed as neurobiological models of complex systems displaying maladaptive plasticity. PMID- 19219301 TI - New nitric oxide donors based on ruthenium complexes. AB - Nitric oxide (NO) donors produce NO-related activity when applied to biological systems. Among its diverse functions, NO has been implicated in vascular smooth muscle relaxation. Despite the great importance of NO in biological systems, its pharmacological and physiological studies have been limited due to its high reactivity and short half-life. In this review we will focus on our recent investigations of nitrosyl ruthenium complexes as NO-delivery agents and their effects on vascular smooth muscle cell relaxation. The high affinity of ruthenium for NO is a marked feature of its chemistry. The main signaling pathway responsible for the vascular relaxation induced by NO involves the activation of soluble guanylyl-cyclase, with subsequent accumulation of cGMP and activation of cGMP-dependent protein kinase. This in turn can activate several proteins such as K+ channels as well as induce vasodilatation by a decrease in cytosolic Ca2+. Oxidative stress and associated oxidative damage are mediators of vascular damage in several cardiovascular diseases, including hypertension. The increased production of the superoxide anion (O2-) by the vascular wall has been observed in different animal models of hypertension. Vascular relaxation to the endogenous NO-related response or to NO released from NO deliverers is impaired in vessels from renal hypertensive (2K-1C) rats. A growing amount of evidence supports the possibility that increased NO inactivation by excess O2- may account for the decreased NO bioavailability and vascular dysfunction in hypertension. PMID- 19219302 TI - Synaptic vesicle pool size, release probability and synaptic depression are sensitive to Ca2+ buffering capacity in the developing rat calyx of Held. AB - The calyx of Held, a specialized synaptic terminal in the medial nucleus of the trapezoid body, undergoes a series of changes during postnatal development that prepares this synapse for reliable high frequency firing. These changes reduce short-term synaptic depression during tetanic stimulation and thereby prevent action potential failures during a stimulus train. We measured presynaptic membrane capacitance changes in calyces from young postnatal day 5-7 (p5-7) or older (p10-12) rat pups to examine the effect of calcium buffer capacity on vesicle pool size and the efficiency of exocytosis. Vesicle pool size was sensitive to the choice and concentration of exogenous Ca2+ buffer, and this sensitivity was much stronger in younger animals. Pool size and exocytosis efficiency in p5-7 calyces were depressed by 0.2 mM EGTA to a greater extent than with 0.05 mM BAPTA, even though BAPTA is a 100-fold faster Ca2+ buffer. However, this was not the case for p10-12 calyces. With 5 mM EGTA, exocytosis efficiency was reduced to a much larger extent in young calyces compared to older calyces. Depression of exocytosis using pairs of 10-ms depolarizations was reduced by 0.2 mM EGTA compared to 0.05 mM BAPTA to a similar extent in both age groups. These results indicate a developmentally regulated heterogeneity in the sensitivity of different vesicle pools to Ca2+ buffer capacity. We propose that, during development, a population of vesicles that are tightly coupled to Ca2+ channels expands at the expense of vesicles more distant from Ca2+ channels. PMID- 19219303 TI - Antidipsogenic effects of central adenosine-5'-triphosphate. AB - Besides other physiological functions, adenosine-5'-triphosphate (ATP) is also a neurotransmitter that acts on purinergic receptors. In spite of the presence of purinergic receptors in forebrain areas involved with fluid-electrolyte balance, the effect of ATP on water intake has not been investigated. Therefore, we studied the effects of intracerebroventricular (icv) injections of ATP (100, 200 and 300 nmol/microL) alone or combined with DPCPX or PPADS (P1 and P2 purinergic antagonists, respectively, 25 nmol/microL) on water intake induced by water deprivation. In addition, the effect of icv ATP was also tested on water intake induced by intragastric load of 12% NaCl (2 mL/rat), acute treatment with the diuretic/natriuretic furosemide (20 mg/kg), icv angiotensin II (50 ng/microL) or icv carbachol (a cholinergic agonist, 4 nmol/microL), on sodium depletion-induced 1.8% NaCl intake, and on food intake induced by food deprivation. Male Holtzman rats (280-320 g, N = 7-11) had cannulas implanted into the lateral ventricle. Icv ATP (300 nmol/microL) reduced water intake induced by water deprivation (13.1 +/- 1.9 vs saline: 19.0 +/- 1.4 mL/2 h; P < 0.05), an effect blocked by pre-treatment with PPADS, but not DPCPX. Icv ATP also reduced water intake induced by NaCl intragastric load (5.6 +/- 0.9 vs saline: 10.3 +/- 1.4 mL/2 h; P < 0.05), acute furosemide treatment (0.5 +/- 0.2 vs saline: 2.3 +/- 0.6 mL/15 min; P < 0.05), and icv angiotensin II (2.2 +/- 0.8 vs saline: 10.4 +/- 2.0 mL/2 h; P < 0.05), without changing icv carbachol-induced water intake, sodium depletion-induced 1.8% NaCl intake and food deprivation-induced food intake. These data suggest that central ATP, acting on purinergic P2 receptors, reduces water intake induced by intracellular and extracellular dehydration. PMID- 19219304 TI - GABA in the central amygdaloid nucleus modulates the electrolyte excretion and hormonal responses to blood volume expansion in rats. AB - We investigated the involvement of GABAergic mechanisms of the central amygdaloid nucleus (CeA) in unanesthetized rats subjected to acute isotonic or hypertonic blood volume expansion (BVE). Male Wistar rats bearing cannulas unilaterally implanted in the CeA were treated with vehicle, muscimol (0.2 nmol/0.2 microL) or bicuculline (1.6 nmol/0.2 microL) in the CeA, followed by isotonic or hypertonic BVE (0.15 or 0.3 M NaCl, 2 mL/100 g body weight over 1 min). The vehicle-treated group showed an increase in sodium excretion, urinary volume, plasma oxytocin (OT), and atrial natriuretic peptide (ANP) levels compared to control rats. Muscimol reduced the effects of BVE on sodium excretion (isotonic: 2.4 +/- 0.3 vs vehicle: 4.8 +/- 0.2 and hypertonic: 4.0 +/- 0.7 vs vehicle: 8.7 +/- 0.6 microEq.100 g-1.40 min-1); urinary volume after hypertonic BVE (83.8 +/- 10 vs vehicle: 255.6 +/- 16.5 microL.100 g-1.40 min-1); plasma OT levels (isotonic: 15.3 +/- 0.6 vs vehicle: 19.3 +/- 1 and hypertonic: 26.5 +/- 2.6 vs vehicle: 48 +/- 3 pg/mL), and ANP levels (isotonic: 97 +/- 12.8 vs vehicle: 258.3 +/- 28.1 and hypertonic: 160 +/- 14.6 vs vehicle: 318 +/- 16.3 pg/mL). Bicuculline reduced the effects of isotonic or hypertonic BVE on urinary volume and ANP levels compared to vehicle-treated rats. However, bicuculline enhanced the effects of hypertonic BVE on plasma OT levels. These data suggest that CeA GABAergic mechanisms are involved in the control of ANP and OT secretion, as well as in sodium and water excretion in response to isotonic or hypertonic blood volume expansion. PMID- 19219305 TI - A simple model for circadian timing by mammals. AB - Circadian timing is structured in such a way as to receive information from the external and internal environments, and its function is the timing organization of the physiological and behavioral processes in a circadian pattern. In mammals, the circadian timing system consists of a group of structures, which includes the suprachiasmatic nucleus (SCN), the intergeniculate leaflet and the pineal gland. Neuron groups working as a biological pacemaker are found in the SCN, forming a biological master clock. We present here a simple model for the circadian timing system of mammals, which is able to reproduce two fundamental characteristics of biological rhythms: the endogenous generation of pulses and synchronization with the light-dark cycle. In this model, the biological pacemaker of the SCN was modeled as a set of 1000 homogeneously distributed coupled oscillators with long range coupling forming a spherical lattice. The characteristics of the oscillator set were defined taking into account the Kuramoto's oscillator dynamics, but we used a new method for estimating the equilibrium order parameter. Simultaneous activities of the excitatory and inhibitory synapses on the elements of the circadian timing circuit at each instant were modeled by specific equations for synaptic events. All simulation programs were written in Fortran 77, compiled and run on PC DOS computers. Our model exhibited responses in agreement with physiological patterns. The values of output frequency of the oscillator system (maximal value of 3.9 Hz) were of the order of magnitude of the firing frequencies recorded in suprachiasmatic neurons of rodents in vivo and in vitro (from 1.8 to 5.4 Hz). PMID- 19219306 TI - L-histidine enhances learning in stressed zebrafish. AB - The aim of the present study was to determine the effect of the histaminergic precursor L-histidine and the H3 receptor antagonist thioperamide on the learning process of zebrafish submitted or not to confinement stress. On each of the 5 consecutive days of experiment (D1, D2, D3, D4, D5), animals had to associate an interruption of the aquarium air supply with food offering. Non-stressed zebrafish received an intraperitoneal injection of 100 mg/kg L-histidine, 10 mg/kg thioperamide or saline after training. Stressed animals received drug treatment and then were submitted to confinement stress for 1 h before the learning procedure. Time to approach the feeder was measured (in seconds) and was considered to be indicative of learning. A decrease in time to approach the feeder was observed in the saline-treated group (D1 = 141.92 +/- 13.57; D3 = 55 +/- 13.54), indicating learning. A delay in learning of stressed animals treated with saline was observed (D1 = 217.5 +/- 25.66). L-histidine facilitated learning in stressed (D1 = 118.68 +/- 13.9; D2 = 45.88 +/- 8.2) and non-stressed (D1 = 151.11 +/- 19.20; D5 = 62 +/- 14.68) animals. Thioperamide inhibited learning in non-stressed (D1 = 110.38 +/- 9.49; D4 = 58.79 +/- 16.83) and stressed animals (D1 = 167.3 +/- 26.39; D5 = 172.15 +/- 27.35). L-histidine prevented the increase in blood glucose after one session of confinement (L-histidine = 65.88 +/- 4.50; control = 53 +/- 3.50 mg/dL). These results suggest that the histaminergic system enhances learning and modulates stress responses in zebrafish. PMID- 19219307 TI - L-histidine reduces inhibitory avoidance in Carassius auratus submitted to cerebellar ablation. AB - The effect of post-training treatment with L-histidine (LH) on the memory consolidation of inhibitory avoidance was investigated in Carassius auratus submitted to cerebellar ablation. The inhibitory avoidance procedure included 3 days: one habituation day, one training day (5 trials, T1-T5) and one test day. On the training day, each fish was placed individually in a white compartment separated from a black compartment by a sliding door. When the fish crossed into the black compartment, a weight was dropped in front of it (aversive stimulus) and the time to cross was recorded. Saline or LH (100 mg/kg) was injected intraperitoneally 10 min after the trials. Data were log10 transformed and analyzed by ANOVA and the Student-Newman-Keuls test (P < 0.05). In T5, all groups [ablation/LH (N = 15; 189.60 +/- 32.52), ablation/saline (N = 14; 204.29 +/- 28.95), sham/LH (N = 14; 232.36 +/- 28.15), and sham/saline (N = 15; 249.07 +/- 25.82)] had similar latencies that were significantly higher than T1 latencies [ablation/LH (89.33 +/- 20.41), ablation/saline (97.00 +/- 25.16), sham/LH (73.86 +/- 18.42), and sham/saline (56.71 +/- 17.59)], suggesting acquisition of inhibitory avoidance. For the test, there was a significant reduction in latencies of ablation/LH (61.53 +/- 17.70) and sham/saline (52.79 +/- 25.37) groups compared to the ablation/saline (213.64 +/- 29.57) and sham/LH (199.43 +/- 24.48) groups, showing that cerebellum ablation facilitated retention of inhibitory avoidance and LH reversed the effect of ablation. The results support other evidence that LH impairs memory consolidation and/or reduces the interpretation of aversion value. PMID- 19219308 TI - Antinociception synergy between the peripheral and spinal sites of the heme oxygenase-carbon monoxide pathway. AB - We have shown that the peripheral and spinal cord heme oxygenase (HO)-carbon monoxide (CO)-soluble guanylate cyclase-cGMP pathways play an important role in antinociception in the rat experimental formalin model. Our objective was to determine if there is synergism between peripheral (paw) and spinal HO-CO pathways in nociception. Rats were handled and adapted to the experimental environment for a few days before the formalin test, in which 50 microL of a 1% formalin was injected subcutaneously into the dorsal surface of the right hind paw. The animals were then observed for 1 h and the frequency of flinching behavior was taken to represent the nociceptive response. Thirty minutes before the test, rats were pretreated with intrathecal injections of the HO inhibitor, zinc deuteroporphyrin 2,4-bis glycol (ZnDPBG) or heme-lysinate, which is a substrate of the HO pathway. The paw treatments took place 20 min before the test. Low doses of ZnDPBG did not increase nociception, while a low heme-lysinate dose did not change flinching behavior after paw or spinal injections. Combined subactive spinal (50 nmol) and peripheral (40 nmol) low doses of ZnDPBG induced hypernociception (increase of 80% in the first and 25% in the second phase flinching), whereas combined spinal-peripheral heme-lysinate (50 and 30 nmol) led to second phase antinociception (40% reduction in flinching). These findings suggest a synergy between the peripheral and spinal HO-CO pathways. Local activation of the HO system probably regulates the nociception initiation in peripheral tissue and participates in buffering the emerging nociceptive signals at the peripheral and spinal sites of action. In short, an antinociceptive synergy exists between peripheral and spinal HO pathways, which may reduce the doses required and side effects. PMID- 19219309 TI - In the February 2009 issue of Clinics. PMID- 19219310 TI - Correlations between the stomatognathic system and body posture: biological or clinical implications? PMID- 19219311 TI - Antiphospholipid antibodies and multiple organ failure in critically ill cancer patients. AB - OBJECTIVES: To describe the clinical outcomes and thrombotic events in a series of critically ill cancer patients positive for antiphospholipid (aPL) antibodies. DESIGN: Retrospective case series study. SETTING: Medical-surgical oncologic intensive care unit (ICU). PATIENTS AND PARTICIPANTS: Eighteen patients with SIRS/sepsis and multiple organ failure (MOF) and positive for aPL antibodies, included over a 10-month period. INTERVENTIONS: None MEASUREMENTS AND RESULTS: aPL antibodies and coagulation parameters were measured up to 48 hours after the occurrence of acrocyanosis or arterial/venous thrombotic events. When current criteria for the diagnosis of aPL syndrome were applied, 16 patients met the criteria for 'probable' and two patients had a definite diagnosis of APL syndrome in its catastrophic form (CAPS). Acrocyanosis, arterial events and venous thrombosis were present in eighteen, nine and five patients, respectively. Sepsis, cancer and major surgery were the main precipitating factors. All patients developed MOF during the ICU stay, with a hospital mortality rate of 72% (13/18). Five patients were discharged from the hospital. There were three survivors at 90 days of follow-up. New measurements of lupus anticoagulant (LAC) antibodies were performed in these three survivors and one patient still tested positive for these antibodies. CONCLUSIONS: In this small series of patients, we observed a high frequency of auto-antibodies and micro- and macro-vascular thrombotic events in critically ill cancer patients. The coexistence of sepsis or SIRS and aPL antibodies was often associated with MOF and death. More studies are necessary to determine the pathophysiological significance of antiphospholipid antibodies in severely ill cancer patients. PMID- 19219312 TI - Validation of the Brazilian-Portuguese version of the Gesture Behavior Test for patients with non-specific chronic low back pain. AB - OBJECTIVE: To develop a Brazilian version of the gesture behavior test (GBT) for patients with chronic low back pain. METHODS: Translation of GBT into Portuguese was performed by a rheumatologist fluent in the language of origin (French) and skilled in the validation of questionnaires. This translated version was back translated into French by a native-speaking teacher of the language. The two translators then created a final consensual version in Portuguese. Cultural adaptation was carried out by two rheumatologists, one educated patient and the native-speaking French teacher. Thirty patients with chronic low back pain and fifteen healthcare professionals involved in the education of patients with low back pain through back schools (gold-standard) were evaluated. Reproducibility was initially tested by two observers (inter-observer); the procedures were also videotaped for later evaluation by one of the observers (intra-observer). For construct validation, we compared patients' scores against the scores of the healthcare professionals. RESULTS: Modifications were made to the GBT for cultural reasons. The Spearman's correlation coefficient and the intra-class coefficient, which was employed to measure reproducibility, ranged between 0.87 and 0.99 and 0.94 to 0.99, respectively (p < 0.01). With regard to validation, the Mann-Whitney test revealed a significant difference (p < 0.01) between the averages for healthcare professionals (26.60; SD 2.79) and patients (16.30; SD 6.39). There was a positive correlation between the GBT score and the score on the Roland Morris Disability Questionnaire (r= 0.47). CONCLUSIONS: The Brazilian version of the GBT proved to be a reproducible and valid instrument. In addition, according to the questionnaire results, more disabled patients exhibited more protective gesture behavior related to low-back. PMID- 19219313 TI - Selective fetoscopic laser photocoagulation of superficial placental anastomoses for the treatment of severe twin-twin transfusion syndrome. AB - OBJECTIVE: To report our initial institutional experience with fetoscopic laser photocoagulation of placental anastomoses in severe twin-twin transfusion syndrome using a 1.0 mm endoscope. METHODS: Between July 2006 and June 2008, 19 monochorionic diamniotic twin pregnancies complicated by severe TTTS (Quintero stages III and IV) underwent fetoscopic laser therapy. Perinatal data were prospectively collected and compared according to the Quintero stages. RESULTS: Nine patients were classified as stage III and ten as stage IV. The Mean gestational ages at diagnosis and procedure were 20 (range: 17-25) and 22.0 (range: 19.0-26.0) weeks, respectively, with no statistical difference between the two groups. Preterm premature rupture of the membranes occurred in two cases (10.5%), and spontaneous preterm delivery in eight (42.1%). Overall mean gestational age at delivery was 32.1 (range: 26.0-38.0) weeks. Prematurity was more severe in stage IV patients (p<0.01). Among all cases, the overall survival rate was 52.6%, and the percentages of pregnancies with survival of both babies and at least one twin were 26.3% and 78.9%, respectively. In the case of stage III patients, the overall survival rate was 61.1%. Of the stage III pregnancies, 33.3% resulted in both babies surviving, and 88.9% of these pregnancies resulted in at least one surviving twin. For stage IV, as the corresponding statistics were 45.0%, 20.0% and 70.0% respectively. CONCLUSIONS: Our initial institutional experience with 1.0 mm fetoscopic laser therapy for severe TTTS showed results similar to those reported in the literature for larger endoscopes. PMID- 19219314 TI - Comparison of radial shockwaves and conventional physiotherapy for treating plantar fasciitis. AB - OBJECTIVE: To compare radial shockwave treatment and conventional physiotherapy for plantar fasciitis. MATERIALS AND METHODS: Thirty-two patients with plantar fasciitis were included in this study. They were randomly divided into two groups. Group 1 was composed of 16 patients who underwent 10 physiotherapy sessions each, consisting of ultrasound, kinesiotherapy and instruction for stretching exercises at home. Group 2 was composed of 16 patients who underwent three applications of radial shockwaves (once a week) and received instruction for stretching exercises at home. Pain and ability to function were evaluated before treatment, immediately afterwards, and three months later. The mean age of the patients was 47.3 +/- 10.3 years (range 25-68); 81% were female, 87% were overweight, 56% had bilateral impairment, and 75% used analgesics regularly. RESULTS: Both treatments were effective for pain reduction and for improving the functional abilities of patients with plantar fasciitis. The effect of the shockwaves was apparent sooner than physiotherapy after the onset of treatment. CONCLUSION: Shockwave treatment was no more effective than conventional physiotherapy treatment when evaluated three months after the end of treatment. PMID- 19219315 TI - Interaction between intra-abdominal pressure and positive-end expiratory pressure. AB - OBJECTIVE: The aim of this study was to quantify the interaction between increased intra-abdominal pressure and Positive-End Expiratory Pressure. METHODS: In 30 mechanically ventilated ICU patients with a fixed tidal volume, respiratory system plateau and abdominal pressure were measured at a Positive-End Expiratory Pressure level of zero and 10 cm H2O. The measurements were repeated after placing a 5 kg weight on the patients' belly. RESULTS: After the addition of 5 kg to the patients' belly at zero Positive-End Expiratory Pressure, both intra abdominal pressure (p<0.001) and plateau pressures (p=0.005) increased significantly. Increasing the Positive-End Expiratory Pressure levels from zero to 10 cm H2O without weight on the belly did not result in any increase in intra abdominal pressure (p=0.165). However, plateau pressures increased significantly (p< 0.001). Increasing Positive-End Expiratory Pressure from zero to 10 cm H2O and adding 5 kg to the belly increased intra-abdominal pressure from 8.7 to 16.8 (p<0.001) and plateau pressure from 18.26 to 27.2 (p<0.001). Maintaining Positive End Expiratory Pressure at 10 cm H2O and placing 5 kg on the belly increased intra-abdominal pressure from 12.3 +/- 1.7 to 16.8 +/- 1.7 (p<0.001) but did not increase plateau pressure (26.6+/-1.2 to 27.2 +/-1.1 -p=0.83). CONCLUSIONS: The addition of a 5 kg weight onto the abdomen significantly increased both IAP and the airway plateau pressure, confirming that intra-abdominal hypertension elevates the plateau pressure. However, plateau pressure alone cannot be considered a good indicator for the detection of elevated intra-abdominal pressure in patients under mechanical ventilation using PEEP. In these patients, the intra-abdominal pressure must also be measured. PMID- 19219316 TI - Plantar pressure distribution patterns during gait in diabetic neuropathy patients with a history of foot ulcers. AB - OBJECTIVE: To investigate and compare the influence of a previous history of foot ulcers on plantar pressure variables during gait of patients with diabetic neuropathy. INTRODUCTION: Foot ulcers may be an indicator of worsening diabetic neuropathy. However, the behavior of plantar pressure patterns over time and during the progression of neuropathy, especially in patients who have a clinical history of foot ulcers, is still unclear. METHODS: Subjects were divided into the following groups: control group, 20 subjects; diabetic neuropathy patients without foot ulcers, 17 subjects; and diabetic neuropathy patients with at least one healed foot ulcer within the last year, 10 subjects. Plantar pressure distribution was recorded during barefoot gait using the Pedar-X system. RESULTS: Neuropathic subjects from both the diabetic neuropathy and DNU groups showed higher plantar pressure than control subjects. At midfoot, the peak pressure was significantly different among all groups: control group (139.4+/-76.4 kPa), diabetic neuropathy (205.3+/-118.6 kPa) and DNU (290.7+/-151.5 kPa) (p=0.008). The pressure-time integral was significantly higher in the ulcerated neuropathic groups at midfoot (CG: 37.3+/-11.4 kPa.s; DN: 43.3+/-9.1 kPa.s; DNU: 68.7+/-36.5 kPa.s; p=0.002) and rearfoot (CG: 83.3+/-21.2 kPa.s; DN: 94.9+/-29.4 kPa.s; DNU: 102.5+/-37.9 kPa.s; p=0.048). CONCLUSION: A history of foot ulcers in the clinical history of diabetic neuropathy subjects influenced plantar pressure distribution, resulting in an increased load under the midfoot and rearfoot and an increase in the variability of plantar pressure during barefoot gait. The progression of diabetic neuropathy was not found to influence plantar pressure distribution. PMID- 19219317 TI - Comparison of self-report and interview administration methods based on the Brazilian versions of the Western Ontario Rotator Cuff Index and Disabilities of the Arm, Shoulder and Hand Questionnaire in patients with rotator cuff disorders. AB - OBJECTIVE: The purpose of the present study was to compare self-report and interview administration methods using the Western Ontario Rotator Cuff Index (WORC) and Disabilities of the Arm, Shoulder and Hand Questionnaire (DASH) in patients with rotator cuff disorders. METHODS: Thirty male and female patients over 18 years of age with rotator cuff disorders (tendinopathy or rotator cuff tear) and Brazilian Portuguese as their primary language were recruited for assessment via administration of the Western Ontario Rotator Cuff Index and and Disabilities of the Arm, Shoulder and Hand Questionnaire. A randomization method was used to determine whether the questionnaires would be self-reported (n=15) or administered by an interviewer (n=15). Pearson correlation coefficients were used to evaluate the correlation between the Western Ontario Rotator Cuff Index and and Disabilities of the Arm, Shoulder and Hand Questionnaire in each group. The t test was used to determine whether the difference in mean questionnaire scores and administration time was statistically significant. For statistical analysis, the level of significance was set at 5%. RESULTS: The mean subject age was 55.07 years, ranging from 27 to 74 years. Most patients had a diagnosis of tendinopathy (n=21). With regard to level of schooling, the majority (n=26) of subjects had completed a college degree or higher. The mean questionnaire scores and administration times did not significantly differ between the two groups (p>0.05). There were statistically significant correlations (p<0.05) between Western Ontario Rotator Cuff Index and and Disabilities of the Arm, Shoulder and Hand Questionnaire, and strong correlations were found between the questionnaires in both groups. CONCLUSION: There are no differences between the Western Ontario Rotator Cuff Index and Disabilities of the Arm, Shoulder and Hand Questionnaire administration methods with regard to administration time or correlations between the questionnaires. PMID- 19219318 TI - Registry of Hospital das Clinicas of the University of Sao Paulo Medical School: first official solid organ and tissue transplantation report - 2008. AB - OBJECTIVE: The aim of this study was to report a single center experience of organ and tissue transplantation INTRODUCTION: This is the first report of organ and tissue transplantation at the Hospital das Clinicas of the University of Sao Paulo Medical School. METHODS: We collected data from each type of organ transplantation from 2002 to 2007. The data collected were patient characteristics and actuarial survival Kaplan-Meier curves at 30 days, one year, and five years RESULTS: There were a total of 3,321 transplants at our institution and the 5-year survival curve ranged from 53% to 88%. CONCLUSION: This report shows that solid organ and tissue transplants are feasible within the institution and allow us to expect that the quality of transplantation will improve in the future. PMID- 19219319 TI - The prevalence of thyroid dysfunction in elderly cardiology patients with mild excessive iodine intake in the urban area of Sao Paulo. AB - OBJECTIVES: To evaluate the prevalence of thyroid dysfunction in elderly cardiac patients in an outpatient setting. SUBJECTS AND METHODS: A total of 399 consecutive patients (268 women, age range 60-92 years) who were followed at Heart Institute were evaluated for thyroid dysfunction with serum free T4, TSH, anti-Peroxidase antibodies, urinary iodine excretion measurements and thyroid ultrasound. RESULTS: Hyperthyroidism (overt and subclinical) was present in 29 patients (6.5%), whereas hypothyroidism (overt and subclinical) was found in 32 individuals (8.1%). Cysts were detected in 11 patients (2.8%), single nodules were detected in 102 (25.6%), and multinodular goiters were detected in 34 (8.5%). Hashimoto's thyroiditis was present in 16.8% patients, most of whom were women (83.6%). The serum TSH increased with age and was significantly higher (p= <0.01) in patients, compared to the normal control group. No significant differences in serum TSH and free T4 values were observed when patients with atrial fibrillation (AF) where compared with those without arrhythmia. The median urinary iodine levels were 210 microg/L (40-856 microg/L), and iodine levels were higher in men than in women (p<0.01). Excessive iodine intake (urinary iodine >300 microg/L) was observed in one-third of patients (30.8%). CONCLUSIONS: Elderly patients have a higher prevalence of both hypo- and hyperthyroidism as well as thyroid nodules when compared with the general population. About one third of the older patients had elevated urinary secretion of iodine and a higher prevalence of chronic Hashimoto's thyroiditis. It is recommended that ultrasonographic studies, tests for thyroid function and autoimmunity should be evaluated in elderly patients. PMID- 19219321 TI - Treatment of recurrent mandibular myxoma by curettage and cryotherapy after thirty years. PMID- 19219320 TI - Reduction of postsurgical adhesions in a rat model: a comparative study. AB - BACKGROUND: Adhesion formation after peritoneal surgery is a major cause of postoperative bowel obstruction, infertility, and chronic pelvic pain. In this study, we compared the possible individual effects of phosphatidylcholine (PC), Seprafilm II, and tissue plasminogen activator (t-PA) and the combined effects of phosphatidylcholine and t-PA on postoperative adhesion formation in a rat surgical model. MATERIALS AND METHODS: A total of 50 Wistar male rats underwent median laparotomy and standardized abrasion of the visceral and parietal peritoneum. phosphatidylcholine, Seprafilm II, and t-PA alone and phosphatidylcholine and t-PA in combination were applied intraperitoneally at the end of the surgical procedure. Seven days after surgery, a relaparotomy was performed for adhesion grading and histopathological examination. RESULTS: A comparison of adhesion stages demonstrated a significant difference between the control group and the study groups (p<0.001). The adhesion grade of the combined treatment group was statistically different from that of the other groups (p<0.05). In the t-PA group and the combined group, six and two rats, respectively, developed hematomas locally on the cecum. CONCLUSIONS: PC, t-PA, and Seprafilm II used individually reduced the adhesion grade. The t-PA and phosphatidylcholine combination was most effective in reducing adhesion formation. On the other hand, usage of t-PA alone or in combination may increase risk of bleeding. More detailed studies are needed, and future studies on the efficacy of a material for decreasing adhesion formation should include a comparison of several control materials in the same model. PMID- 19219322 TI - A rare quadruplicate arrangement of abductor pollicis longus tendons: anatomical and clinical relevance. PMID- 19219323 TI - Osteomalacia and vitamin D deficiency in the elderly. PMID- 19219324 TI - Surgical treatment of emphysema. PMID- 19219325 TI - Prevalence of asthma in children and adolescents in a city in the Brazilian Amazon region. AB - OBJECTIVE: To analyze the prevalence of asthma and asthma symptoms in students of two distinct age brackets residing in the city of Tangara da Serra, Brazil. METHODS: Cross-sectional, population-based study of the prevalence of asthma in children from 6 to 7 years of age and adolescents from 13 to 14, using the standardized International Study of Asthma and Allergies in Childhood, phase 1 questionnaire, validated for use in Brazil. Students who responded affirmatively to question 2 (presence of wheezing in the preceding 12 months) were classified as suffering from asthma. RESULTS: The study comprised 3,362 students, of whom 1,634 (48.6%) were children and 1,728 (51.4%) were adolescents. Of the 1,634 children, 816 (49.9%) were male, and 818 (50.1%) were female. Of the 1,728 adolescents, 773 (45.0%) were male, and 955 (55.0%) were female. The prevalence of asthma among the children was 25.2%, whereas that among the adolescents was 15.9% (chi2 = 8.34; p = 0.00). The children presented higher prevalences of the following symptoms of asthma than did the adolescents: wheezing ever (54.3%), nocturnal dry cough (43.9%), wheezing in the preceding 12 months (25.2%), and from 1 to 3 attacks of wheezing in the preceding 12 months (19.1%). There were no differences between the two groups regarding physician-diagnosed asthma (approximately 4.5%). There were no statistical differences regarding the prevalence of asthma by gender in the two groups. CONCLUSIONS: Tangara da Serra has a high prevalence of asthma in children and adolescents, and this result is compatible with other studies carried out in Brazil and Latin America using the same methodology. PMID- 19219326 TI - Therapeutic application of collateral ventilation with pulmonary drainage in the treatment of diffuse emphysema: report of the first three cases. AB - OBJECTIVE: To report the results obtained in three patients with diffuse pulmonary emphysema during the pre- and post-operative periods following a new surgical technique: collateral ventilation with lung parenchyma drainage. METHODS: Patients suffering from pulmonary failure and disabling dyspnea, despite having received the gold standard treatment, including pulmonary rehabilitation, were selected for the evaluation of pulmonary drainage. During the pre- and post operative periods, patients were submitted to plethysmography and six-minute walk tests, as well as completing the following quality of life questionnaires: Medical Outcomes Study 36-item Short-Form Health Survey, Saint George's Respiratory Questionnaire, Eastern Cooperative Oncology Group Performance Status and Medical Research Council Scale. In all three cases, the postoperative follow up period was at least 300 days. The tests were performed at the following time points: during the pre-operative period; between post-operative days 30 and 40; and on post-operative day 300. Data were analyzed using profile plots of the means. RESULTS: When the results obtained in the pre-operative period were compared to those obtained at the two post-operative time points evaluated, improvements were observed in all parameters studied. CONCLUSIONS: The results suggest that the surgical technique proposed for the treatment of patients suffering from severe diffuse emphysema successfully reduces the debilitating symptoms of these patients, improving their quality of life considerably. PMID- 19219327 TI - Influence of lean body mass on cardiopulmonary repercussions during the six minute walk test in patients with COPD. AB - OBJECTIVE: Although lean body mass (LBM) has been associated with mortality in patients with COPD, its influence on functional limitation is not clear. The objective of this study was to analyze the cardiopulmonary variables in COPD patients with or without LBM depletion, prior to and after the six-minute walk test (6MWT). METHODS: We evaluated COPD patients, 32 with LBM depletion and 36 without. All patients underwent clinical evaluation, spirometry, evaluation of body mass composition and 6MWT, as well as completing questionnaires related to quality of life and perception of dyspnea. RESULTS: No significant differences in the severity of airway obstruction, perception of dyspnea and quality of life scores were found between the groups. The distance covered on the 6MWT was similar in COPD patients with and without LBM depletion (470.3 +/- 68.5 m vs. 448.2 +/- 89.2 m). However, patients with LBM depletion presented significantly greater differences between baseline and final values in terms of heart rate and Borg scale index for lower limb fatigue. There was a significant positive correlation between distance covered on the 6MWT and FEV1 (r = 0.381, p = 0.01). CONCLUSIONS: In the patients studied, functional exercise tolerance and quality of life were unaffected by LBM depletion. However, the patients with LBM depletion presented more pronounced lower limb fatigue during the 6MWT, which underscores the importance of the evaluation and treatment of systemic manifestations in COPD patients. PMID- 19219328 TI - Perception of disease severity in adult patients with cystic fibrosis. AB - OBJECTIVE: To evaluate the perception of disease severity in patients with cystic fibrosis (CF), investigating its relationship with clinical score, radiographic score, respiratory function tests, adherence to treatment and perception of self care practices. METHODS: Prospective, cross-sectional study involving CF patients treated in a program for adults with CF. The perception of disease severity, adherence to treatment and reported self-care practices were evaluated by means of questionnaires. Clinical data, Shwachman-Kulczycki clinical score, Brasfield radiographic score and spirometry were obtained for all of the patients. RESULTS: Of the 38 patients studied, 3 (7.9%) patients rated their perception of health status as well below average; 5 (13.2%), as below average; 15 (39.5%), as average; 10 (26.3%), as above average; and 5 (13.2%), as well above average. The perception of disease severity correlated significantly with clinical score (r = 0.43, p = 0.007), FVC (r = 0.34, p = 0.034), FEV1 (r = 0.38, p = 0.019) and self care practices (r = 0.33, p = 0.044), but not with degree of adherence (r = 0.03, p = 0.842) and radiographic score (r = 0.33, p = 0.51). CONCLUSIONS: The perception of disease severity correlated with objective measurements of disease severity (clinical score and respiratory function tests) and with reported self care practices, but not with adherence to treatment. PMID- 19219329 TI - Pulmonary deposition of inhaled tobramycin prior to and after respiratory therapy and use of inhaled albuterol in cystic fibrosis patients colonized with Pseudomonas aeruginosa. AB - OBJECTIVE: To evaluate whether respiratory therapy followed by the use of inhaled albuterol modifies the pulmonary deposition of inhaled tobramycin in patients with cystic fibrosis (CF) and whether pulmonary deposition correlates with disease severity or genotype. METHODS: A prospective study was carried out including patients with CF older than 6 years of age and colonized with Pseudomonas aeruginosa. Exclusion criteria were pulmonary exacerbation, changes in therapy between the study phases and FEV1 < 25%. All patients were submitted to pulmonary scintigraphy by means of a scintillation camera equipped with a low energy all purpose collimator in order to evaluate drug penetration following the administration of inhaled 99mTc-tobramycin, as well as to pulmonary perfusion with 99mTc-macroaggregated albumin (phase 1). One month later, the same procedure was performed following respiratory therapy and administration of inhaled albuterol (phase 2). RESULTS: We included 24 patients (12 males) aged 5-27 years (mean +/- SD: 12.85 +/- 6.64 years). The Shwachman score (SS) was excellent/good in 8 patients, moderate/fair in 16 and poor in 0. Genotyping revealed that 7 patients were DeltaF508 homozygotes, 13 were DeltaF508 heterozygotes; and 4 presented other mutations. In all patients, lung deposition of tobramycin decreased in phase 2, especially in those with moderate/fair SS (p = 0.017) and in heterozygotes (p = 0.043). CONCLUSIONS: The use of a respiratory therapy technique and the administration of inhaled albuterol immediately prior to the use of inhaled tobramycin decreased the pulmonary deposition of the latter in CF patients, and this reduction correlates with disease severity and genotype. PMID- 19219330 TI - Prevalence of temporary social security benefits due to respiratory disease in Brazil. AB - OBJECTIVE: To determine the prevalence of temporary social security benefits due to respiratory disease granted to employees, as well as the number of lost workdays and costs resulting from those in Brazil between 2003 and 2004. METHODS: Cross-sectional study using data obtained from the Unified System of Benefits of the Brazilian Institute of Social Security (INSS, Instituto Nacional de Seguro Social) and the Brazilian Social Registry Database. Data regarding gender, age, diagnosis and type of economic activity, as well as type, duration and cost of benefits, were compiled. RESULTS: Respiratory diseases accounted for 1.3% of the total number of temporary social security benefits granted by INSS, with a prevalence rate of 9.92 (per 10,000 employment contracts). Females and individuals older than 50 years of age were the most affected. Non-work-related benefits were more common than were work-related benefits. The most prevalent diseases were pneumonia, asthma and COPD, followed by laryngeal and vocal cord diseases. The most prevalent types of economic activity were auxiliary transportation equipment manufacturing, tobacco product manufacturing and computer-related activities. The mean duration of benefits was 209.68 days, with a mean cost of R$ 4,495.30 per occurrence. Respiratory diseases caused by exogenous agents demanded longer sick leave (mean, 296.72 days) and greater cost (mean, R$ 7,105.74). CONCLUSIONS: The most prevalent diseases were airway diseases and pneumonia. Workers from auxiliary transportation equipment manufacturing, tobacco product manufacturing and computer-related activities were the most affected. Diseases caused by exogenous agents demanded longer sick leaves and resulted in greater costs. PMID- 19219331 TI - Clinical evolution of a group of patients with multidrug-resistant TB treated at a referral center in the city of Rio de Janeiro, Brazil. AB - OBJECTIVE: To analyze the clinical characteristics and evolution of a group of patients with positive sputum cultures for multidrug-resistant (MDR) Mycobacterium tuberculosis and treated at a referral center in the city of Rio de Janeiro, Brazil. METHODS: Based on the positive results in sputum cultures for MDR M. tuberculosis, 50 patients were selected, and their clinical data were obtained from the Brazilian Ministry of Health MDR-TB Database. The frequencies of noncompliance, relapses, failures and previous treatments for TB up to diagnosis of MDR-TB were compiled. The radiological patterns were classified as unilateral or bilateral, and with or without cavitation. Two years after the end of the standard treatment for MDR-TB, the outcome (cure, failure, noncompliance or death) for each patient was evaluated and reassessed every two years. The post treatment follow-up period was eight years. RESULTS: The mean number of previous treatments was 2.3 +/- 0.9. The mean interval between the initial diagnosis and the development of MDR-TB was 2.0 +/- 1.7 years. Two years after the initial treatment for MDR-TB, 2 patients had abandoned treatment, 8 had died, 18 had been cured, and 22 had presented treatment failure. The bivariate analysis showed that bilateral pulmonary involvement and cavitary pattern markedly reduced the chances for cure, with a relative risk of 1-0.6 (40%) and 1-0.7 (30%), respectively. At the end of the follow-up period, 2 patients had abandoned treatment, 9 had presented treatment failure, 17 had been cured, and 22 had died. CONCLUSIONS: Bilateral pulmonary involvement and cavity pattern greatly reduced the chances for cure of the patients with MDR-TB. Most patients who presented treatment failure died within the 8-year follow-up period. PMID- 19219332 TI - Prevalence of active and latent TB among inmates in a prison hospital in Bahia, Brazil. AB - OBJECTIVE: To determine the prevalence of latent and active TB among detainees in a prison hospital in Bahia, Brazil. METHODS: A cross-sectional study with prospective data collection was carried out comprising 237 inmates in the Bahia State Prison Hospital between July 2003 and April 2004. A standardized questionnaire was applied and completed by medical students. The detainees were systematically submitted to the following tests: tuberculin skin test, chest X ray (anteroposterior), sputum smear microscopy and culture for mycobacteria. The events of interest were active TB and latent TB. RESULTS: The mean age of the participants was 36.6 years, and 89.9% were male. Smoking and alcohol consumption were reported by 70.0% and 43.9% of the inmates, respectively. A history of treatment for TB was reported by 11.3% of the inmates. Of the inmates evaluated, 36.3% reported cough and 31.4% reported expectoration. Other less common symptoms were asthenia (in 26.2%), weight loss (in 23.1%), loss of appetite (in 17.7%), fever (in 11.3%) and hemoptysis (in 6.7%). Of the 86 inmates tested, none presented positive HIV serology. The prevalence of latent TB was 61.5% (96 of the 156 inmates submitted to tuberculin skin tests), whereas that of active TB was 2.5% (6 of the 237 inmates evaluated). The presence of cough was a determinant of active TB (prevalence ratio = 8.8; 95% CI: 1.04-73.9; p = 0.025). CONCLUSIONS: Active and latent TB are highly prevalent among inmates hospitalized in the Bahia State Prison Hospital. Our findings justify the need to implement public policies specifically directed towards the control of TB in this population. PMID- 19219333 TI - Design and evaluation of a device for collecting exhaled breath condensate. AB - In recent years, the analysis of exhaled breath condensate samples has been given great weight as a noninvasive methodology of studying physiology and lung diseases. The present study describes a device for measuring exhaled breath condensate that is affordable, easily constructed, portable and suitable for use in the field, as well as allowing the collection of simultaneous samples. The results obtained with this device in terms of the concentrations of pH, peroxide oxide and nitrite, metabolites related to inflammatory and oxidative damage, in exhaled breath condensate samples are comparable to those obtained with other devices previously described. PMID- 19219334 TI - Psychometric properties of the Fagerstrom Test for Nicotine Dependence. AB - OBJECTIVE: The Fagerstrom Test for Nicotine Dependence (FTND) is a screening instrument for physical nicotine dependence and is extensively used in various countries. The objective of the present report was to review articles related to the psychometric properties of the FTND. METHODS: A systematic search for articles published up through December of 2007 was carried out in various electronic databases. The following search terms were used: 'Fagerstrom Test for Nicotine Dependence'; 'FTND'; 'psychometric'; 'validity'; 'reliability'; 'feasibility'; and 'factors'. We included articles published in English, Spanish or Portuguese and in which the psychometric properties of the FTND were evaluated. RESULTS: Twenty-six studies related to the psychometric properties of the FTND were identified in the indexed literature. Analysis of the studies confirmed the reliability of the FTND for the assessment of nicotine dependence in different settings and populations. CONCLUSIONS: Further validation studies using previously validated instruments as a comparative measure are needed before the extensive use of the FTND can be justified on the basis of its psychometric qualities. PMID- 19219335 TI - Tuberculosis in health professionals: a new perspective on an old problem. AB - The objective of this review was to contribute to the debate on the nosocomial transmission of TB among health professionals in a country where TB is endemic. Prior to 1900, there was no reason to believe that health professionals interacting with TB patients were more susceptible to becoming infected with the bacillus than was the general population. Between 1920 and 1930, various studies showed significant findings regarding the rates of positive tuberculin skin tests among students in the area of health care. However, most clinicians remained skeptical about the susceptibility of health professionals to becoming infected with TB. In the various locales where the treatment of patients with TB has been implemented, health professionals have been described as an especially predisposed population to becoming infected with and developing active TB. It is urgent that the scientific community and health professionals become mobilized, recognizing themselves as a population at risk of developing TB, and that actions be taken in order to minimize the potential risks of acquiring the disease at locales where patients with TB are treated. PMID- 19219336 TI - Repeat pulmonary thromboendarterectomy after recurrence of chronic thromboembolic pulmonary hypertension. AB - Pulmonary thromboendarterectomy has been established as the standard method for the treatment of chronic thromboembolic pulmonary hypertension, with excellent results. However, repeat pulmonary thromboendarterectomy due to recurrence of pulmonary embolism has never been reported in the Brazilian literature. Its safety and effectiveness remain obscure. We report the case of a patient presenting recurrence of chronic thromboembolic pulmonary hypertension five years after the first pulmonary thromboendarterectomy and requiring a second operation for resolution of the symptoms. PMID- 19219337 TI - Chronic necrotizing pulmonary aspergillosis. AB - Chronic necrotizing pulmonary aspergillosis is one of the forms of pulmonary aspergillosis typically found in mildly immunocompromised patients. We report the case of a female patient with complaints of chronic productive cough, fever and asthenia. She reported previous corticosteroid use. A CT scan of the chest revealed consolidation with interposed cavitation in the right upper lobe. Fiberoptic bronchoscopy revealed purulent fluid within the tracheobronchial tree and an endobronchial exophytic lesion. The results of the biopsy of that lesion and the transbronchial biopsy were consistent with aspergillosis. Based on the clinical, radiological and histopathological findings, the patient was diagnosed with chronic necrotizing pulmonary aspergillosis. Treated with itraconazole, the patient presented a favorable clinical-radiological evolution. PMID- 19219338 TI - Extralobar pulmonary sequestration with hemothorax secondary to pulmonary infarction. AB - Pulmonary sequestration is an uncommon condition that accounts for 0.5-6% of all pulmonary malformations and is typically diagnosed in childhood. Of the two forms of pulmonary sequestration, intralobar and extralobar, the latter is less frequently encountered. The current report describes the case of a 32-year-old female patient with chest and abdominal pain. Imaging (chest X-rays and CT scans of the chest) revealed consolidation and pleural effusion. The initial thoracocentesis revealed hemothorax. Subsequent diagnostic video-assisted thoracoscopy revealed extralobar pulmonary sequestration. Consequently, the therapeutic decision was to make the conversion to thoracotomy in order to resect the lesion and safely ligate the intercostal vascular pedicle. PMID- 19219339 TI - [Risk factors and assistance to dyslipidemia in climacteric women]. PMID- 19219340 TI - [Factors associated with women's dyslipidemia in the post-menopause]. AB - PURPOSE: to evaluate factors associated with women's dyslipidemia during menopause. METHODS: case-control study of prevalent cases and controls selected from a dedicated outpatient clinic. From recent biochemical parameters found in patients' files, women have been grouped in 'case' and 'control'. Women who presented any alteration in the blood levels of total cholesterol, LDL cholesterol, triglycerides and/or HDL-cholesterol were considered as case, and the ones who presented normal levels of them, as control. Data concerning socioeconomic situation, physical activity, etilism and tabagism, anthropometric measurements and food ingestion have been collected and then compared between the groups. Ratios have been compared by the chi2, Fisher's exact test and/or t Student test, depending on the distribution type. The crude relationship between each factor and the presence of dyslipidemia has been estimated by logistic regression. RESULTS: data have been collected from 84 women aged from 42 to 59 years, as 45 of them were grouped as case (dyslipidemic) and 39 as control (non dyslipidemic). Age average of cases and controls was 52.1+/-4.2 and 52.2+/-4.7 years old, respectively. The sample showed to be homogeneous for the socioeconomic characteristics (income, occupation and schooling), physical activity practice, etilism and tabagism, and food ingestion, with no significant correlation with dyslipidemia. The groups presented an income up to two minimal wages, low schooling level (up to the fourth grade of lower school), and the housewife occupation. Smoking and drinking alcohol was not very frequent. Practicing physical activity was non-existent, thus characterizing a sedentary population. Food ingestion was adequate for carbohydrates, protein, lipids, but not for cholesterol (excessive) and fibers (insufficient), in both groups. Concerning the anthropometric evaluation, there has been an association with dyslipidemia, as the body mass index (BMI) and the waist circumference (WC) were significantly larger in case than in control. The waist-hip ratio has been similar in both groups. Weight excess has been found in most of the cases (73.3%) as almost half of them (44%) presented WC >88 cm, which represents a very increased risk. CONCLUSIONS: it is possible to conclude that, in the studied sample, only the anthropometric measurements have been considered as risk factors associated with dyslipidemia, during post-menopause. PMID- 19219341 TI - [Importance of a second opinion in breast surgical pathology and therapeutic implications]. AB - PURPOSE: to evaluate the agreement between histopathologic diagnoses of breast lesions made by general pathologists and by a specialist in breast pathology. METHODS: a cohort retrospective study comparing histopathologic diagnoses of 329 cases of breast lesions received in consultation for a second opinion was carried out. The material received for consultation included slides (152 cases), paraffin blocks (59 cases) or slides and blocks (118 cases). Cases were reviewed and the original diagnoses and diagnoses from a specialist in breast pathology were compared. The main diagnoses, nuclear grade of ductal carcinoma in situ, and the histopathologic grade of invasive mammary carcinomas were evaluated. The kappa index and percentual concordance were used in the statistical analyses. RESULTS: a moderate agreement was observed between the original histopathologic diagnoses and the second opinion (kappa index=0.48; percentual concordance=59.9%). The diagnosis of malignancy was confirmed in 185/225 cases (82.2%) and diagnosis of benign lesions was confirmed in 89/104 cases (85.6%). The highest agreement was observed in the diagnosis of invasive mammary carcinomas (81%) and the highest disagreement was observed among diagnoses of ductal carcinoma in situ with microinvasion (74%), lobular carcinoma in situ (70%), and atypical epithelial hyperplasias (61%). There was a moderate agreement in the nuclear grade of ductal carcinoma in situ (kappa index=0.52; percentual concordance=68.8%), and good concordance in the histologic grade of invasive carcinomas (kappa index=0.61; percentual concordance=74.3). CONCLUSIONS: the results show higher concordance rate in the diagnosis of invasive carcinomas and lower concordance in the diagnosis of ductal carcinoma in situ with microinvasion and premalignant breast lesions, especially lobular neoplasia in situ, and atypical epithelial hyperplasias. PMID- 19219342 TI - [Predictive factors for height gain in idiopathic central precocious puberty treated with GnRH analogues]. AB - PURPOSE: to evaluate predictive factors of response to GnRHa treatment in girls with idiopathic central precocious puberty. METHODS: a retrospective cohort study was conducted involving 33 girls diagnosed with idiopathic central precocious puberty and treated with GnRHa. The following independent variables were assessed: age at the beginning of therapy and at the onset of symptoms, time elapsed since the appearance of pubertal characteristics and the beginning of treatment, bone age, bone age advance, duration of GnRHa treatment, actual height and Z-score, predicted height and Z-score and hormone measurements of FSH and LH after GnRH stimulation, which were correlated with gain in height as a dependent variable at treatment discontinuation, calculated by the difference between the predicted height at the end and beginning of treatment. For statistical analysis, Pearson's linear correlation was used, in addition to multiple linear regression analysis. RESULTS: the mean age at the beginning of treatment was 7.8+/-1.3 years, with a mean bone age of 10.1+/-1.6 years. Bone age advance was 2.3+/-1.1 years and was controlled during the treatment period. Gain in predicted height was 2.5+/-1.3cm. It was positively correlated with time elapsed since the beginning of symptoms and the beginning of treatment and with bone age advance, while negatively correlated with the Z-score of height at the beginning of treatment and predicted height at the beginning of treatment, and the latter was the main factor determining gain from treatment. CONCLUSIONS: girls who had the most significant compromise of predicted adult height, as detected by a larger deviation from the population (Z-score) and the most considerable advance in bone age, received benefit from GnRHa therapy, and they must not be excluded from the group to be treated. PMID- 19219343 TI - [Prevalence of Chlamydia and Neisseria gonorrhoeae infections in pregnant women in six Brazilian cities]. AB - PURPOSE: This paper has aimed at estimating the prevalence of infections by Chlamydia trachomatis and by Neisseria gonorrhoeae in pregnant women from six Brazilian cities, identifying its association with socio-economical and demographic variables. METHODS: This study has been part of a multicentric nationwide transversal research, with samples of pregnant women attended from 2004 to 2005 in basic attention pre-natal services from six Brazilian cities (Manaus, Fortaleza, Goiania, Rio de Janeiro, Sao Paulo and Porto Alegre). Cervico vaginal samples have been collected from all the pregnant women, and have afterwards been submitted to the hybrid capture technique in order to identify chlamydia and gonococcus. Socio-demographic, medical, sexual and obstetric information have been collected through specific questionnaires. The Odds Ratio (OR) has been used to evaluate risk factors associated to infection by gonorrhea and chlamydia. Statistical analysis has been done with the t-Student, chi2 and Fisher's exact tests. RESULTS: Three thousand and three pregnant women with an average age of 23.8 years old (+/-6.9) took part in the study. Infection prevalence by chlamydia and gonococcus were 9.4 and 1.5, respectively. Ten per cent of the pregnant women with chlamydia have presented gonococcus simultaneously. The risk of presenting one of those infections was two times higher for the women under 20. The infection main predictors have been: age under 20, race/black, single/separated and report of over one partner in the previous year. CONCLUSIONS: This study has observed high prevalence of infection by Chlamydia trachomatis and by Neisseria gonorrhoeae in Brazilian pregnant women. The main risk factor for the infection has been to be under 20 years old. PMID- 19219344 TI - [Teenage pregnancy outcome]. AB - PURPOSE: to compare delivery and pregnancy follow-up among adolescent and non adolescent pregnant women whose delivery occurred in a tertiary hospital from Regiao de Lisboa (Portugal). METHODS: retrospective study with 10,656 deliveries. Pregnancy follow-up, delivery type, need of episiotomy and severe lacerations, Apgar index at the fifth minute and the delivery weight have been evaluated. The pregnant women were divided into two groups, over and under 20 years old. The group with women under 20 was further subdivided in pregnant women under or over 16. The chi2 test has been used for statistical analysis. RESULTS: adolescents presented worse follow-up: first appointment after 12 weeks (46.4 versus 26.3%) and less than four appointments (8.1 versus 3.1%), less dystocia (21.5 versus 35.1%), less caesarian sections (10.6 versus 20.7%), and lower need for inducing labor (16.5 versus 26.5%). There was no significant difference concerning gestational age at delivery and ratio of low weight newborns. Among adolescents, the ones under 16 had more low weight newborns (12 versus 7.4%) and more deliveries between 34 and 37 weeks (10.8 versus 4.2%). CONCLUSIONS: in a hospital attending adolescents with social and psychological support, the fact of them having had a worse follow-up in the pre-natal phase, their performance has not been worse. Nevertheless, special attention might be given to pregnant women under 16. PMID- 19219345 TI - [Lymphoma of the uterine cervix: report of two cases and review of the literature]. AB - The occurrence of primary lymphomas of the female genital tract is rare. The diagnosis is usually not possible by the cytological examination; a tissue biopsy is necessary. The present paper reports two patients referred to our service with a diagnosis of cervical lymphoma. In one of them, the diagnostic difficulties are demonstrated. Both patients were submitted to chemotherapy with satisfactory post operatory period. There is no standard treatment for uterine lymphomas. Exclusive local treatment is supported by some reports in the literature in clinical stage IE, while others prefer systemic treatment irrespective of clinical stage. PMID- 19219346 TI - [Sepsis and septic shock during pregnancy: clinical management]. AB - Sepsis is one of the main causes of maternal death, being related to infections from obstetric origin (infected abortion, chorioamnionitis, puerperal infection) or non-obstetric (resulting from infections which occur in other areas). This review aims at describing the mechanisms involved in the physiopathology of this entity and at updating the clinical approach to sepsis, recommended in international guidelines (early goal-directed therapy--precocious resuscitation, or precocious treatment guided by goals), as well as at calling attention to the influence of pregnancy both in the clinical manifestation and in the therapeutic management of septic conditions. PMID- 19219347 TI - [REBEn: increasing the contribution]. PMID- 19219348 TI - [The relation between the nursing team and patient's family: implications for care]. AB - Study of qualitative nature, developed along with the nursing team and the accompanying relatives of interned customers in a public hospital at Vitoria/ES. The objectives were to describe the bases that sustain the relations among those subjects; to analyze the dynamics of this relation; and to discuss its implications for the nursing care. The theoretical reference was linked to the studies of power relations in the disciplined hospital environment and the care in a humanized perspective concept. The data collection was through a participant observation and semi-structured interviews. The results pointed out that those relations don't happen lineally. At same time that conflicting experiences emerge, there is another whose position of one and other subject reveals sensibility and solidarity that implicate directly in the delivered care to the hospitalized customer. PMID- 19219349 TI - [The feeding habits of adolescents from public schools in Fortaleza, CE, Brazil]. AB - The research aimed at learning food habits of adolescents from public schools in Fortaleza, CE, Brazil. Seven hundred students age ranging from 4 to 19 years-old were interviewed from twelve education institutions by means of a structured form. From the usual food consumed the energetic rice and bread were highlighted, consumed by 95.8% and 85.2% of the adolescents, respectively (p=0.0001). Within constructor food, meat and beans are ingested by 60.6% and 75.0%, respectively (p=0.0001). Regulative food like fruit and vegetables do not have a usual consumption among adolescents, since just (34.3%) and (47.6%) respectively consume them (p=0.0001 and p=0.226). An unbalance in proper nutrient ingestion for the adolescence was stated, favoring overweight and, consequently, chronic diseases like mellitus diabetes. PMID- 19219350 TI - [Diagnosed with Parkinson's diesease: impact on patients and family members]. AB - This was a qualitative nature, developed with the objective of understanding the impact of a Parkinson's diagnosis on the individual and the family. Data were collected between October and December 2007, from 20 individuals residing in Maringa, PR - 10 Parkinson's patients and their closest family member - using semi-structured interviews. From the analysis of the data, two themes emerged: 'Coming to terms as a Parkinson's patient: a long path' and 'Difficulties and feeling after the diagnosis', which reveal that the lack of knowledge on the disease is widespread among the general population and that the quality of the first contact with the diagnosis is crucial in the determination and attitude of individuals toward the illness. PMID- 19219352 TI - [Disease-related absenteeism among nursing team members in a teaching hospital]. AB - The article presents results of the research that identified to causes of disease related absenteeism, justified by medical license, among nursing employees of a public hospital in Montes Claros, MG, Brazil. This is a descriptive-exploratory study. Results indicate that the main reasons for absenteeism are related to musculoskeletal problems and to respiratory diseases; mainly between married female nurse aides, with age between 25 and 45 years, that works in day shift and with more than one employment. It was concluded that there is a necessity to develop prevention actions for controlling the absenteeism as a strategy for the quality of life of nursing workers. PMID- 19219353 TI - [The institutional violence in day-care and pre-schools according to mothers' view]. AB - The objective of this research is to know the institutional violence in the day care centers and preschools by the mothers' view. It was elaborated with two approaches:quantitative and qualitative. The researcher through interview and observation made the collection of primary data, and the qualitative data werw collected by recorded interview, being made the content analysis. The results show that 23,9%of the mothers affirm that violence exists against the child; the proportion of children that didn't suffer violence in the day-care center can be considered statistically significant, when cmpared with those already suffered, independently of the time of frequency in the day-care center (p= 0,95);the mothers say they don't know the cause of the institutional violence; the physical violence appears with larger incidence; it is more frequent the mother to try to know the cause of theviolence, and the minus, to denounce to Guardian Council. It's compatible to the parents the child's protection and they need to know the ways of protecting them of the infantile violence. PMID- 19219351 TI - [The main caregiver of oncologic terminally ill patient, repercussions of this task]. AB - OBJECTIVES: to identify the profile of the caregivers of terminally ill patient, to inquire the activities, changes and difficulties. The researchers applied a questionnaire. From all the 53 caregivers, 48 were female. 92% of the caregivers were relative to the patient. Despite of the financial problems, their relationship between caregiver and patient improved. The hygiene cares, feeding, medication were responsibility of the caregiver. The activities developed increased their amount of job, negative physical and psychical repercussions appeared. 60% of the caregivers answered that the most convenient place of the patient's death was the hospital, 40% chose home. There is a distinction between the routine of care and the death of the patient; the caregivers in fact take care of life. PMID- 19219354 TI - [Urinary incontinence and aging: impact on daily basis and on the quality of life]. AB - Human aging is accompanied by worn out of the physical functions of body and mind as well as a decrease in physiological response to the environment. Consequently these change the quality of life for those going through the aging process, especially regarding independence and autonomy. Among these common aging changes, there is the urinary incontinence having multiple etiologies with great therapeutic complexity, which creates a huge impact on the life quality of these people. This article will focus on part of a research done for a master's dissertation from one of the authors, which sought to understand in what proportion urinary incontinence affects, not only their daily lives but also their quality of life. PMID- 19219355 TI - [Determinatives of family caregiver's tension while caring the dependent elderly]. AB - This study had as objective investigate the determinatives of the tension of the dependent elderly's family caregiver. Its accomplish happened in the domestic scenery, involving thirty principal caregivers. The data were collected by a recorded and estructured interview. The analyses of the objective data was made by means of a quantitative approach. The discursive data, for being collected through a structured instrument, dispensed analytic procedure, being distribute between questions that referee in the way of amply your comprehension. The results appoint that the investigated phenomenon starts from determinatives related to the caregiver, to the elderly's deficits, to the interection elderly/caregiver, to the environment, and related to the care demands. PMID- 19219356 TI - [Constructing community involvement for the prevention of home accidents in children]. AB - INTRODUCTION: This paper reports part of a study whose aim was to mobilise a team for the Family Health Program to enable the citizen community involvement aiming at preventing domestic accidents for children under age five, through the theoretical framework of the Seven Theses by Roberto Briceno-Leon. METHODOLOGY: the Convergent Care Research was used with a Team for the Family Health Programme in a town in the country in Rio Grande do Sul. Educational workshops were first performed with the team and then educational activities were done with the families in the community. Data were collected through the observation, interview and content analysis. RESULTS: Creation of four educational tools: the song 'A Licao do Sapeca', dramatisation of the song, the practice called 'O que e isto?' and the practice called 'Casa Simulada'. CONSIDERATIONS: It is showed that through creativity and congruence the health team unleashed a process of the citizen community involvement for preventing child domestic accidents. PMID- 19219357 TI - [Risk for vascular trauma: diagnosis proposal and validation by experts]. AB - This concept and content validation study aimed to compose the title, vulnerability mechanisms and risk factors for the nursing diagnosis 'Risk for Vascular Trauma', according to the taxonomy proposed by NANDA, as well as to validate the elements of this diagnosis, based on 60 experts' opinion. Fifty-one factors were identified, related to five axes: medication and infusion form/periodicity; intravascular catheter and dwelling in the same site; fixation of the intravascular catheter; individuals, their habits, communication standard, lifestyle and sensory-motor ability; and professional decisions, institutional policy and procedures. Among these, 28 vulnerability factors were identified (score > 0.80). The results evidenced that 'Risk for Vascular Trauma' is a new nursing diagnosis, and its clinical validation is suggested. PMID- 19219358 TI - [Concepts of a multidisciplinar team on runaway patients]. AB - A qualitative study that aimed at learning the way an emergency care multidisciplinary team at a public university hospital conceives and deals with the problem of patients who run away from hospital was carried out. Data were collected using the focus groups technique, whose sample was constituted by ten individuals, approached by non-directive group dynamics. Three thematic classes resulted from content analysis: a certain ambivalence; emergency, a place of (lack of) control; fears and insecurities. The study indicates the chaotic situation encountered by emergency services and, in this context, when patients run away, the health care team is imminently subject to a triple judgment: social, legal and institutional. PMID- 19219359 TI - [Nursing education and the challenges for health promotion]. AB - This is a descriptive-exploratory study with qualitative approach whose objective was analyze the health promotion on nursing education. The subjects were teachers, students & professionals whom receive the students on the scenarios of learningship of two courses of graduation in Minas Gerais/Brazil. The results indicate the incorporation of health promotion as a referencing for nursing formation favored by schoolservice articulation as well as an important strategy. In the mean time, it needs major valorization as a condition to maintain the changes occurred in nursing formation. It concludes that is necessary strategic actions for change the sanitary and education practices involving the 'good' practices of health promotion on overcoming the challenges to change the nursing education. PMID- 19219360 TI - [Nursing diagnoses for patients with Acquired Immunodeficiency Syndrome]. AB - This study aims at identifying the most frequent nursing diagnoses, according to the NANDA's taxonomy II in adult patients of HIV/AIDS and to check how often the defining characteristics, the related factors and the risk factors provided support for their development. Data survey was conducted by researchers in the 60 patients admitted with HIV/aids, followed by the finding out whether the NANDA's proposed requirements to develop the Nursing Diagnoses were appropriate. From the 38 diagnoses, 13 diagnoses were chosen with a frequency equal or higher than 80%. Results pointed to agreement and appropriateness for identification. But new studies will be necessary either to acknowledge the findings and bring new knowledge to the topic as well, besides encouraging research with this patients' population. PMID- 19219361 TI - [Religion and spirituality: the experience of families of children with Chronic Renal Failure]. AB - OBJECTIVE: To describe the manifestations of religiosity and spirituality in families of children with Chronic Kidney Failure undergoing peritoneal dialysis. METHODS: This qualitative research is a case study with multiple subjects. Meaningful content analysis was adopted, using the thematic type, in the framework of family nursing in health promotion. RESULTS: Four families participated in the study, totaling fourteen participants. Religion and spirituality appear as important resources for family members in coping with the chronic disease, mainly in view of threatening prognoses. CONCLUSION: It is important for nurses to understand the family's religious and spirituality in the disease process, with a view to their work in health promotion. PMID- 19219362 TI - [Palliative care to adolescents with cancer: a literature review]. AB - Providing care to adolescents with cancer in the process of death and dying has been a great challenge for health professionals. This challenge is marked by a high emotional burden and specificities of this stage of human development. The purpose of the present study was to review the scientific literature regarding palliative care to adolescents with cancer. This study is a literature review, which data collection was performed using Lilacs, Medline, and PsycInfo, in addition to non-systematic databases. An analysis of the manuscripts revealed three themes: adolescence and its different definitions; the particularities of adolescents with cancer; and palliative care to adolescents with cancer. The study showed there is a scarcity of evidenced-based research defining the panorama of symptoms affecting the quality of life during palliative care and an absence of specific programs in the stage of fast changes that, alone, demand for adaptive efforts. PMID- 19219363 TI - [Family health: challenges in the reorientation process of the assistance model]. AB - The Family Health Program (PSF) constitutes a strategy for reformulating the health assistance model in Brazil. This study was carried out to ponder over the challenges of this strategy concerning to health practices in the primary care context. After existing for more than twelve years, there appear questionings related to the paper of PSF in the reformulation of the way to think and promote health, and their structures remain permeable to the traditional hegemonic model that corrupts the daily work process. Some changes and reconstruction are necessary in the health practices concerning to care production based on light, relational and integrate technologies that are centered on users' needs. Although the deconstruction/reconstruction process implicates much labor, the advantage in overcoming the challenges faced by PSF rather leads to the transformation of reality and to construction of health practices that are solidary, welcoming, and resolvent. PMID- 19219364 TI - [HIV and the female gender: the psychological experience]. AB - AIDS raises a social issue, humanitarian, legal, and threatens to change the social structure founding of tolerance and understanding, that sustains the life of our societies. From a clinical and epidemiological background of HIV/AIDS, this article of literature review makes an approach about psychological experience and implication of the Human Immunodeficiency Virus infection in pregnant woman. PMID- 19219365 TI - [Accreditation processes for nurses: a historical briefing]. AB - To identify, describe and analyze the historical milestones of nurses' accreditation in the critical care area, a research was carried out at LILACS and MEDLINE, unearthing 16 publications, led to the 4 categories. In the USA, the CCNE and NLNAC focus on evaluating Nursing courses and educational institutions, whilst generalist nurses are evaluated by NCLEX, and experts are evaluated by professional associations. Accrediting nurses is also a matter of interest in Spain, Canada, South Africa and Australia. In the USA, the process began in the 40's, whilst in Spain it began in the 90's. It was concluded that the professional certification and accreditation in Nursing occur mainly in developed countries and began in the middle of the last century. PMID- 19219366 TI - [Medication: ethical and legal aspects for nursing]. AB - This article treats of the duties and responsibilities of nursing staff concerning medication process. From a survey about the valid legislation, the authors made considerations about the legal implications to the nursing staff, especially related to the assistance quality detours involving the medication process, as well as the contradictions in law that restrict the information access about medication to these professionals. PMID- 19219367 TI - [Leadership in critical care settings: reflections and challenges for the Brazilian Nursing]. AB - The objective of this study was to reflect on the leadership, especially in the intensive care nursing, based on the prospect of nurses, teachers, nursing students and in the literature on the subject. The leadership of the nurse in intensive care units faces challenges and paradoxes and is present in the planning of care, in the moments of ethical and bioethical conflicts, in the supervision of care, in capacity of making decisions quickly and safety, in knowing deal with the issues involving the process of living and dying. PMID- 19219368 TI - [Caring moment: moment of reflection in action]. AB - Reflection-in-action means to think what we do while we are doing. The present text analyzes this activity on the part of the nurse professional considering the caring moment, which constitutes the encounter between the caregiver and the cared for. It is suggested that the reflective practice be stimulated during the educational process and for that, faculty must be prepared. The reflective practice presents advantages such as: updating, renewal, knowledge generation, among others. PMID- 19219369 TI - [Nursing students' experience in a research group]. AB - This is an experience report intended to highlight the importance of the nursing students' participation in the Research Group - Educare - Passo Fundo University, RS, Brasil, and its contribution for the professional qualification. The article reveals the group contributions to the nursing students through its effective participation during the whole research process. It emphasizes this participation as an opportunity for the close relation of theoretical knowledge and the essential practice which is considered an important instrument for the development of creative power intended to search for solutions to solve the problems found in the reality. PMID- 19219370 TI - [Distance education in the Discipline of Legislation, Ethics and Practice Nursing]. AB - Report of experience whose aim is to describe the integration of the discipline of Legislation, Ethics and Practice of Nursing online in the Nursing Course at the University of Estacio Sa, Rio de Janeiro, Brazil. The construction of interactivity in distance education was critical to the continued planning of virtual environment through the development of students' feasible abilities and skills for the theching strategy of Course Political Educational Project combining with the current legislation needs of nursing profession. The development of education articulated by avaliable tools in the virtual environment aims at promoting the critical-relexive development of the future nurse. PMID- 19219371 TI - [Revista Annaes de Enfermagem: nurses' publications about pediatrics (1932 1941)]. AB - Historic-social study whose object was the intellectual production of nurses and students about pediatric nursing in the journal Annaes de Enfermagem, in the period 1932-1941. The primary source refers to the issues of the journal Annaes de Enfermagem considering the established time limits for the study, as well as reports and correspondences. The secondary sources are constituted by books, articles, dissertations and thesis on the Brazilian history of nursing. On data analysis it was used the thought of the French sociologist Pierre Bourdieu. The results show that the journal Annaes de Enfermagem published important issues of nursing care to the child and contributed to the visibility of Brazilian nurses among the scientific community. PMID- 19219372 TI - Metabolic syndrome may not be a good predictor of coronary artery disease in the Iranian population: population-specific definitions are required. AB - The inability of two definitions of metabolic syndrome (MS) to predict the presence of coronary artery disease (CAD) in a non-Caucasian population was demonstrated. Angiography was used as the gold standard method for identifying patients with (cases) or without (controls) CAD. It is possible that the current definitions of MS are not appropriate for some non-Caucasian populations. PMID- 19219373 TI - Anaplastic sarcoma of the kidney. AB - Wilms tumor can appear with a wide spectrum of morphologic features and can sometimes cover or delay the recognition of other clinicopathologic entities of the kidney. We present a case of a new tumor entity of the kidney, namely the anaplastic sarcoma of the kidney, a tumor of high malignancy. PMID- 19219374 TI - Association between HIV status and positive prostate biopsy in a study of US veterans. AB - It is estimated that 1.2 million Americans are infected with HIV. With advances in treatment and improved survival, HIV-positive patients are increasingly reaching an age when prostate cancer becomes a health issue. While there have been some reports in the literature reporting lower incidence of prostate cancer in HIV population cohorts, these studies have focused on younger populations, where we expect a lower incidence of prostate cancer. Our study involves patients over a 5.5 year period from a busy referral Veterans Medical Center referred to our urology clinic with either elevated PSA or abnormal DRE. Of these patients referred to our clinic, there is a markedly higher rate of prostate cancer in HIV patients when compared to our HIV-negative or HIV-unknown population. Though one may surmise a referral bias, in our highly regulated system, we use identical referral criterion for both HIV-positive and HIV-negative patients. Though this is a study with admittedly limited numbers, we believe this report is important because it is one of the first studies to address prostate cancer in a older cohort of patients who are referred because of suspicion for prostate cancer. PMID- 19219375 TI - Thermal analysis of whole bacterial cells exposed to potassium permanganate using differential scanning calorimetry: a biphasic dose-dependent response to stress. AB - The DSC technique applied to the study of the toxic impact of permanganate on bacterial cell culture detected the lack of linearity in the dose-response effect. The results were confirmed by the traditional assay of colony forming ability. The changed pattern of thermal spectra of A. oxydans at permanganate treatment, the measurement of the total heat capacity and the temperature of DNP complex demonstrate the possibility to verify the toxic impact in dependence of concentrations value. PMID- 19219376 TI - Characterization of a PLLA-collagen I blend nanofiber scaffold with respect to growth and osteogenic differentiation of human mesenchymal stem cells. AB - We developed a nanofiber scaffold by blending PLLA with collagen I, suitable for bone regeneration. Among several PLLA-COLI ratios tested, cell growth was better enhanced when blends with a ratio of PLLA-COLI 4:1 were used. Here, growth as well as osteoblast differentiation of hMSC was improved when compared to PLLA nanofibers alone. Therefore, blending is a suitable tool to enhance PLLA nanofibers with respect to bone tissue engineering. PMID- 19219377 TI - Mature miR-184 and squamous cell carcinoma of the tongue. AB - Human microRNA 184 (miR-184) is overexpressed in squamous cell carcinoma (SCC) of the tongue. In vitro inhibition of miR-184 levels could induce apoptosis and hinder proliferation of tongue SCC cells. Patients with tongue SCC have high plasma miR-184 levels. Plasma miR-184 is likely associated with the tumor load. Surgical removal of the primary tumor reduced plasma miR-184 levels significantly. The data suggested that miR-184 is linked to the pathogenesis of tongue SCC. Further studies are warranted to evaluate the use of microRNA-based serological markers in monitoring tongue SCC. PMID- 19219378 TI - The N-terminal domain of thrombospondin-1: a key for the dual effect of TSP-1 in angiogenesis and cancer progression? AB - The data presented in this article suggest that intact TSP-1 may act, in normal vessel homeostasis, as an angiostatic factor favoring vessel quiescence, or even vessel regression, but this activity would be largely impaired in the presence of an excess of angiogenic stimuli and proteases. The fact that the N-terminal domain of TSP-1 (heparin-binding domain or HBD) is recognized by a plethora of cell receptors, all of them engaged in proangiogenic responses, strongly suggests that the proteolytic cleavage of HBD may be relevant in certain pathophysiological conditions. PMID- 19219379 TI - Correlation between changes in ultrasound measurements and clinical curative effect of tension-free vaginal tape-SECUR* procedure. AB - INTRODUCTION AND HYPOTHESIS: The hypotheses of this study were that the TVT-SECUR procedure restricts urethral mobility, which leads to a greater likelihood of curative effect, and that the restriction might change over time. METHODS: Analyses of the position of the urethra and the tape of 85 patients who underwent the TVT-S procedure were performed using perineal ultrasonography. The efficacy of the TVT-S procedure was evaluated by cough test and by the questionnaire ICIQ UI SF. RESULTS: Objectively, 53/85 (62%) of patients had a negative cough test, and in 32/85 (38%) of patients leakage of urine persisted. The TVT-S procedure restricts urethral mobility, and a higher degree of obstruction is associated with a higher likelihood of cure. The restriction weakens within the first 3 months after surgery. CONCLUSIONS: The objective cure rate of TVT-S procedure is low irrespective of placement technique. This may be due to insufficient restriction of urethral mobility. PMID- 19219380 TI - The ethical challenge of surgical innovation. PMID- 19219381 TI - Effect of GLP-1 treatment on bone turnover in normal, type 2 diabetic, and insulin-resistant states. AB - It has been suggested that hormones released after nutrient absorption, such as glucose-dependent insulinotropic peptide (GIP) and glucagon-like peptide 2 (GLP 2), could be responsible for changes in bone resorption. However, information about the role of GLP-1 in this regard is scanty. Diabetes-related bone loss occurs as a consequence of poor control of glucose homeostasis, but the relationship between osteoporosis and type 2 diabetes remains unclear. Since GLP 1 is decreased in the latter condition, we evaluated some bone characteristics in streptozotocin-induced type 2 diabetic (T2D) and fructose-induced insulin resistant (IR) rat models compared to normal (N) and the effect of GLP-1 or saline (control) treatment (3 days by osmotic pump). Blood was taken before and after treatment for plasma measurements; tibiae and femora were collected for gene expression of bone markers (RT-PCR) and structure (microCT) analysis. Compared to N, plasma glucose and insulin were, respectively, higher and lower in T2D; osteocalcin (OC) and tartrate-resistant alkaline phosphatase 5b were lower; phosphate in IR showed a tendency to be higher; PTH was not different in T2D and IR; all parameters were unchanged after GLP-1 infusion. Bone OC, osteoprotegerin (OPG) and RANKL mRNA were lower in T2D and IR; GLP-1 increased OC and OPG in all groups and RANKL in T2D. Compared to N, trabecular bone parameters showed an increased degree of anisotropy in T2D and IR, which was reduced after GLP-1. These findings show an insulin-independent anabolic effect of GLP-1 and suggest that GLP-1 could be a useful therapeutic agent for improving the deficient bone formation and bone structure associated with glucose intolerance. PMID- 19219382 TI - Effects of tumor-induced osteomalacia on the bone mineralization process. AB - Fibroblast growth factor 23 (FGF23) overexpression has been identified as a causative factor for tumor-induced osteomalacia (TIO) characterized by hypophosphatemia due to increased renal phosphate wasting, low 1,25(OH)(2)D(3) serum levels, and low bone density. The effects of long-lasting disturbed phosphate homeostasis on bone mineralization are still not well understood. We report on a patient with a 12-year history of TIO, treated with 1,25(OH)(2)D(3) and phosphate, who finally developed hyperparathyroidism with gland hyperplasia before the tumor could be localized in the scapula and removed. During surgery a transiliac bone biopsy was obtained. FGF23 expression in the tumor cells was confirmed by in situ hybridization. Serum FGF23 levels as measured by ELISA were found to be extremely elevated before and decreased after removal of the tumor. Bone histology/histomorphometry and measurement of bone mineralization density distribution using quantitative backscattered electron imaging were performed on the bone biopsy. The data showed important surface osteoidosis and a slightly increased osteoblast but markedly decreased osteoclast number. The mineralized bone volume (-11%) and mineralized trabecular thickness (-18%) were low. The mean degree of mineralization of the bone matrix (-7%), the most frequent calcium concentration (-4.1%), and the amounts of fully mineralized bone (-40.3%) were distinctly decreased, while the heterogeneity of mineralization (+44.5%) and the areas of primary mineralization (+131.6%) were dramatically increased. We suggest that the elevated levels of FGF23 and/or low phosphate concentrations disturb the mineralization kinetics in vivo without affecting matrix mineralization of pre existing bone packets. PMID- 19219383 TI - Low sit-to-stand performance is associated with low femoral neck bone mineral density in healthy women. AB - Bone mass may be adjusted to control the strains produced by muscular activity. We assessed the relationship between maximum rising strength (MRS), a new measurement of sit-to-stand performance, and femoral neck (FN) bone mineral density (BMD), taking into account possible confounding variables. The study population consisted of 249 healthy women aged 18-76. We measured MRS with a dynamometer fixed on the ground and connected by an adjustable nonelastic cord to a padded belt. FN BMD was measured by dual X-ray absorptiometry. Women in the first quartile of FN BMD (<0.702 g/cm(2)) had significantly lower values of MRS, body weight, height, lean mass, past 5-year physical activity expenditures, blood 17 beta estradiol (E2), 25-hydroxyvitamin D (25(OH)D), dehydroepiandrosterone sulfate (DHEAS), and insulin like growth factor 1, and higher values of age and parathyroid hormone than other women. In the logistic regression model, FN BMD values in the lowest quartile were associated with age (adjusted odds ratio [OR(a)] per 10-year increase = 1.84, 95% confidence interval [95% CI] = 1.33 2.54, P < 0.001), body weight (OR(a) per 10-kg decrease = 3.67, 95% CI = 2.08 6.47, P < 0.001), MRS (OR(a) per 20-kg decrease = 1.17, 95% CI = 1.02-1.34, P = 0.03), serum DHEAS (OR(a) < 0.5 mg/ml vs > or =0.5 mg/ml = 2.83, 95% CI = 1.3 6.12, P = 0. 01), and serum E2 (OR(a) per 10-pmol/l decrease = 1.02, 95% CI = 1.01-1.03, P = 0.03). The present study suggests a significant association between low FN BMD and low sit-to-stand performance in healthy women, independent of possible confounding variables. PMID- 19219385 TI - Mechanical force and cytoplasmic Ca(2+) activate yeast TRPY1 in parallel. AB - The ability to sense mechanical and osmotic stimuli is vital to all organisms from mammals to bacteria. Members of the transient receptor potential (TRP) ion channel family have attracted intense attention for their involvement in mechanosensation. The yeast homologue TRPY1 can clearly be activated by hypertonic shock in vivo and by stretch force under patch clamp. Like its animal counterparts, TRPY1 is polymodal, being gated by membrane stretch force and by cytoplasmic Ca(2+). Here, we investigated how these two gating principles interact. We found that stretch force can induce some channel activation without cytoplasmic Ca(2+). Tens of micromolar Ca(2+) greatly enhance the observed force induced activities, with open probabilities following well the Boltzmann distribution, in which the two gating energies are summed as exponents. To map this formalism to structures, we found Ca(2+)-binding proteins such as calmodulin or calcineurin to be unnecessary. However, removing a dense cluster of negative charges in the C-terminal cytoplasmic domain of TRPY1 greatly diminishes the Ca(2+) activation as well as its influence on force activation. We also found a strategic point upstream of this charge cluster, at which insertion of amino acids weakens Ca(2+) activation considerably but leaves the mechanosensitivity nearly intact. These results led to a structure-function model in which Ca(2+) binding to the cytoplasmic domain and stretching of the membrane-embedded domain both generate gating force, reaching the gate in parallel. PMID- 19219384 TI - Regulation of the Kv2.1 potassium channel by MinK and MiRP1. AB - Kv2.1 is a voltage-gated potassium (Kv) channel alpha-subunit expressed in mammalian heart and brain. MinK-related peptides (MiRPs), encoded by KCNE genes, are single-transmembrane domain ancillary subunits that form complexes with Kv channel alpha-subunits to modify their function. Mutations in human MinK (KCNE1) and MiRP1 (KCNE2) are associated with inherited and acquired forms of long QT syndrome (LQTS). Here, coimmunoprecipitations from rat heart tissue suggested that both MinK and MiRP1 form native cardiac complexes with Kv2.1. In whole-cell voltage-clamp studies of subunits expressed in CHO cells, rat MinK and MiRP1 reduced Kv2.1 current density three- and twofold, respectively; slowed Kv2.1 activation (at +60 mV) two- and threefold, respectively; and slowed Kv2.1 deactivation less than twofold. Human MinK slowed Kv2.1 activation 25%, while human MiRP1 slowed Kv2.1 activation and deactivation twofold. Inherited mutations in human MinK and MiRP1, previously associated with LQTS, were also evaluated. D76N-MinK and S74L-MinK reduced Kv2.1 current density (threefold and 40%, respectively) and slowed deactivation (60% and 80%, respectively). Compared to wild-type human MiRP1-Kv2.1 complexes, channels formed with M54T- or I57T-MiRP1 showed greatly slowed activation (tenfold and fivefold, respectively). The data broaden the potential roles of MinK and MiRP1 in cardiac physiology and support the possibility that inherited mutations in either subunit could contribute to cardiac arrhythmia by multiple mechanisms. PMID- 19219386 TI - Pseudoaneurysm of left ventricular following left ventricular apical venting. PMID- 19219387 TI - Cervical esophageal perforation by transesophageal echocardiography probe detected during coronary artery bypass grafting. PMID- 19219388 TI - Biventricular repair despite hypoplastic tricuspid valve and right ventricle. PMID- 19219389 TI - Transcatheter closure of a ventricular septal defect adjacent to a post infarction aneurysm using an atrial septal defect occluder. PMID- 19219390 TI - Persistent left superior vena cava: an unusual but conquerable obstacle in device implantation. PMID- 19219391 TI - A novel approach to define risk of stent thrombosis after percutaneous coronary intervention with drug-eluting stents: the DERIVATION score. AB - Recent studies of drug-eluting stents (DES) use in routine clinical practice have led to concern regarding their long-term safety and to questions about the adequacy of current antiplatelet therapy guidelines. This study sought to derivate a risk score for predicting stent thrombosis after drug-eluting stenting. The large single center DES Real-world Incremental Value in the erA of percutaneous revascularizaTION (DERIVATION) database, collecting data about 1,377 patients of any age undergoing PCI with DES as treatment for symptomatic coronary artery disease, was use for this purpose. Logistic regression and bootstrap procedure were used to select correlates of stent thrombosis that were subsequently weighted and integrated into an integer scoring system. Five variables selected from the initial multivariate model were weighted proportionally to their respective odds ratio for stent thrombosis [baseline left ventricular ejection fraction <50% (4 points), angioplasty in the setting of acute coronary syndromes (3 points), bifurcation lesion (2 points), left anterior descending as target vessel (2 points), multiple stenting (2 points)]. Three strata of risk were defined (low risk, 0-2; intermediate risk, 3-6; high risk >or=7) with good prognostic accuracy for early, late and very late thrombosis (c statistic = 0.75, 0.65 and 0.73, respectively) in the derivation set. In conclusion, the DERIVATION score may be used as a simple clinical tool for the identification of a sizable cohort in whom close monitoring and aggressive therapy may be beneficial. PMID- 19219392 TI - Significant improvement of right ventricular function by imatinib mesylate in scleroderma-associated pulmonary arterial hypertension. PMID- 19219393 TI - Implantation of the CoreValve self-expanding valve prosthesis via a subclavian artery approach: a case report. PMID- 19219394 TI - Prognostic value of heart rate variability and ventricular arrhythmias during 13 year follow-up in patients with mild to moderate heart failure. AB - BACKGROUND: In contrast to patients with moderate to severe chronic heart failure (CHF), data regarding long-term outcome in patients with mild CHF are scarce. We examined the place of Holter monitoring to study the prognostic value of ventricular arrhythmias and heart rate variability (HRV) in patients with mild to moderate CHF during long-term follow-up. METHODS: We studied 90 patients with mild to moderate CHF and NYHA class II who had been enrolled in the Dutch Ibopamine Multicenter Trial. At baseline their mean age was 60.5 +/- 8.0 years, left ventricular ejection fraction (LVEF) was 0.29 +/- 0.09, and 85% were males. At the start of the study, patients were only using diuretics, while digoxin, and particularly ACE inhibitors and beta-blockers were initiated later. Univariate and multivariate proportional hazard analyses were performed. RESULTS: At baseline 80% of patients were in NYHA class II, and 20% were in class III; their mean age was 60 years, mean LVEF was 0.29, and 85% were men. During a follow-up of 13 years, 47 patients (53%) died. Cardiovascular (CV) death occurred in 39 patients, of which 28 were sudden cardiac death (SCD). For both CV death and SCD, LVEF <30% and ventricular premature beats/h (>20) were independent risk markers. Of the HRV parameters, total power (>2,500 ms(2)) was an important risk marker for CV death, but not for SCD. CONCLUSION: The present 13-year follow-up study in 90 patients with mild to moderate CHF showed that ventricular premature beats and HRV may have important value in predicting outcome. PMID- 19219396 TI - The basic pacing rate in CRT patients: the higher the better? AB - BACKGROUND: To maximize the hemodynamic benefit of cardiac resynchronization therapy (CRT), echocardiographic AV interval optimization is routinely performed, complemented by VV interval optimization especially in non-responders. Programming of the basic pacing rate, however, is largely empirical in these patients. Therefore, the present study aimed to systematically evaluate the impact of basic pacing rate on hemodynamic parameters in CRT patients with sinus bradycardia. METHODS AND RESULTS: We included 70 consecutive patients with moderate to severe heart failure, LV ejection fraction 120 ms combined with echocardiographic evidence of ventricular dyssynchrony. All patients were on optimal heart failure medication, with CRT-ICD devices implanted at least 6 months before inclusion into the study. All patients were in sinus rhythm with a spontaneous heart rate <40 bpm. In all patients, cardiac output (CO) and stroke volume (SV) were determined using electrical velocimetry (EV) (Aesculon, Osypka Medical, Berlin, Germany). EV provides a new algorithm to calculate CO based on variations in thoracic electrical bioimpedance, which has been recently validated. Hemodynamic measurements were performed at four different pacing rates ranging from 40 to 70 bpm. A stepwise increase in CO was encountered with increasing heart rates, reaching statistical significance when comparing 70 with 40 bpm. SV remained unchanged throughout all pacing rates. CONCLUSIONS: In the range between 40 and 70 bpm, an increase in basic pacing rate enhances CO without reducing SV. According to this pilot study, a basic pacing rate between 60 and 70 bpm would appear reasonable. PMID- 19219395 TI - Clinical variables affecting survival in patients with decompensated diastolic versus systolic heart failure. AB - BACKGROUND: The impact of various clinical variables on long-term survival of patients with acutely decompensated diastolic heart failure (DHF) compared to systolic heart failure (SHF) has not been sufficiently investigated. METHODS: Clinical, laboratory, electrocardiographic and echocardiographic data were collected and analyzed for all-cause mortality in 473 furosemide-treated patients aged >or=60 years, hospitalized for acutely decompensated HF. RESULTS: Diastolic heart failure patients (n = 183) were more likely to be older, female, hypertensive, obese, with shorter preexisting HF duration, atrial fibrillation, lower New York Heart Association (NYHA) class, lower maintenance furosemide dosages, and to receive calcium antagonists. The SHF group (290 patients) demonstrated prevailing coronary artery disease, nitrate or digoxin treatment, and electrocardiographic conduction disturbances (P 0.05). Subjects consumed 3,217 +/- 1,027 kcals; 1,848 +/- 768 kcals from carbohydrates; 604 +/- 300 kcals from protein; and 783 +/- 369 kcals from fat. Although work outs differed among one another, within subject training did not vary. These results suggest that measurement of percent body fat in male body builders and weight trainers is equally as accurate using BIA or HW. PMID- 19219399 TI - Hepatic resection in 485 R0 pT2 and pT3 cases of advanced carcinoma of the gallbladder: results of a Japanese Society of Biliary Surgery survey--a multicenter study. AB - PURPOSE: We conducted this study to evaluate the optimal hepatic resection for pT2 and pT3 advanced carcinoma of the gallbladder without invasion of the hepatoduodenal ligament. METHODS: We conducted a questionnaire survey regarding 4,243 cases of carcinoma of the gallbladder treated during the recent 10-year period at 112 institutions belonging to the Japanese Society of Biliary Surgery. The questionnaires included questions on preoperative-diagnosis, complications, treatment, and surgical treatment, procedures of resection, surgical result, path histological findings, mode, and site of recurrence, additional post-operative treatment. They included 293 pT2 and 192 pT3 R0 cases, which were negative for hepatoduodenal ligament invasion, and the cumulative survival rates and sites of postoperative recurrence in the form of liver metastasis, were retrospectively analyzed in these 485 cases. RESULT: There were no significant differences in survival rate or recurrence rates in the form of liver metastasis between the groups that underwent resection of the gallbladder bed, the group that underwent segmentectomy 4a+5, and the group that underwent hepatectomy in patients with of both pT2 or pT3 gallbladder cancers. Our results also did not show that liver metastasis to segment 4a5 alone was particularly common. CONCLUSION: For gallbladder cancer, neither with hepatoduodenal ligament invasion nor hepatic invasion, resection of the gallbladder bed is more preferable for surgical hepatic procedure. For gallbladder cancer that invades any hepatic sites, a hepatic surgical procedure that could eliminate surgical margins would be desirable. PMID- 19219401 TI - The association of circulating monocyte count with coronary collateral growth in patients with diabetes mellitus. AB - The status of inflammation may affect the collateral development in patients with diabetes mellitus (DM). Monocytes were found to have an important role in collateral growth in animal studies. We aimed to investigate the possible association of circulating monocyte count with collateral development in patients with DM and severe coronary artery disease (CAD). We enrolled 134 consecutive patients with DM who had > or =95 stenosis in at least one major coronary artery and investigated the relationship between circulating monocyte count and collateral growth. When we analyzed the coronary angiograms of eligible patients, we found that 64 of them had good collateral growth and 70 had poor collateral growth according to the Cohen-Rentrop method. The monocyte count was significantly different between good and poor collateral growth groups (643 +/- 184 vs. 479 +/- 143 per mm(3), P < 0.001). In the analysis comparing the Rentrop score with the Gensini score and circulating monocyte count, we found significant correlations (r = 0.293, P = 0.001 and r = 0.455, P < 0.001, respectively). The duration of ischemic symptoms tended to be longer in the good collateral group (1.9 +/- 4.1 vs. 0.8 +/- 1.3 years, P = 0.079). The Gensini score was also correlated with the duration of myocardial ischemic symptoms (r = 0.299, P = 0.004). Multivariate analysis revealed an increased monocyte count in the good collateral group [odds ratio (OR), 5.726; 95% confidence interval (CI), 1.817 18.040, P = 0.003, the cut-off value for monocyte was defined as 550 cell/mm(3)]. The increased circulating monocyte count in diabetic patients was evidently related to good coronary collateral growth. This finding may be potentially important in clinical and basic cardiovascular medicine. PMID- 19219402 TI - Identification of sequence changes responsible for the attenuation of avian infectious bronchitis virus strain Arkansas DPI. AB - Infectious bronchitis virus (IBV) is the causal agent of infectious bronchitis, which still remains one of the most important poultry diseases worldwide because of numerous serotypes and variants. A virulent strain of IBV, isolated from Arkansas (Ark), was propagated in embryonated eggs (Ark DPI 11). Following 101 serial passages in embryonated eggs, an attenuated strain of IBV was established (Ark DPI 101) that does not induce histopathological lesions in the tracheae of infected chicks. To identify sequence changes responsible for the attenuation of IBV, complete genome sequences of both virulent and attenuated Ark DPI viruses were obtained. Comparison of the genome sequences of the virulent and attenuated Ark DPI viruses reveals that these viruses are similar and differ only by 21 nucleotides, resulting in 17 amino acids changes. Most of those substitutions are located in the replicase 1a and spike genes. No differences were observed in gene 3, M or 5a, and only one nucleotide substitution each was present in 5b, N and 3'UTR. By comparing the deduced amino acid sequences of virulent and attenuated viruses, we identified sequence changes responsible for the adaptation and attenuation of the IBV-Ark DPI strain. PMID- 19219403 TI - Education differentials by race and ethnicity in the diagnosis and management of hypercholesterolemia: a national sample of U.S. adults (NHANES 1999-2002). AB - OBJECTIVES: To examine education differentials in screening, awareness, treatment and control of hypercholesterolemia overall and in 3 race/ethnic groups. METHODS: We analyzed data for a nationally representative sample of 8,429 men and women ages 20 to 85 years, self-reported as white, black, Mexican American, or other race/ethnicity, who participated in the National Health and Nutrition Examination Survey from 1999-2002. RESULTS: Participants with < high school education were 2.5 times less likely than participants with > or = high school education to have been screened for hypercholesterolemia, after adjusting for age and gender (odds ratio: 0.4, 95 % confidence interval: 0.3-0.5, and similar across race/ethnic group). Multivariable models for awareness, treatment and control showed no significant trends associated with education after adjusting for age, gender, race and comorbidities. CONCLUSIONS: Higher education significantly increased the odds of being screened for hypercholesterolemia overall and within each race/ethnic group. Education differentials were strongest for hypercholesterolemia screening, and weak or no longer apparent for subsequent steps of awareness, treatment and control. Focusing public health policy on increasing screening for individuals with low education might greatly improve their chances of preventing or mitigating morbidity related to hypercholesterolemia and subsequent cardiovascular disease. PMID- 19219404 TI - Dapsone induced methemoglobinemia in a patient with glioblastoma. AB - Primary brain tumor patients have multiple risk factors for Pneumocystis jiroveci and may require prophylaxis with TMP-SMZ or dapsone. Although dapsone is generally safe and efficacious, we present a case of a patient diagnosed with a brain stem glioblastoma who developed methemoglobinemia and haemolytic anemia after presenting with worsening confusion and cardiopulmonary system dysfunction. This case highlights one of the potentially severe complications associated with dapsone therapy. Although this illustrates an unusual toxicity of dapsone, a high index of suspicion should be given to high-risk patients due to ethnic heritage, anemia, or advanced age. Furthermore, given the toxicities of TMP-SMZ and dapsone, further work is needed to determine the threshold CD4(+) count at which empiric prophylaxis should be initiated. PMID- 19219405 TI - Comparison of pain and ecchymosis with low-molecular-weight heparin vs. unfractionated heparin in patients requiring bridging anticoagulation after warfarin interruption: a randomized trial. AB - BACKGROUND: Subcutaneous (SC) low-molecular-weight heparin (LMWH) or unfractionated heparin (UFH) are safe and efficacious for bridging anticoagulation after warfarin interruption. Although LMWH and UFH are self administered by >90% of patients, factors that may be important to patients such as differences in pain and ecchymosis have not been explored. METHODS: We randomized 24 patients to receive SC LMWH or SC UFH twice-daily during the perioperative period. Injection associated pain was recorded using a visual analogue scale and area of ecchymosis was measured by digital photography of the injection site on the day of the procedure. RESULTS: The area of ecchymosis was 2 fold higher with UFH than LMWH (19.4 cm(2) vs. 8.98 cm(2); P = 0.33) and pain was similar with both treatments (115 mm vs. 171 mm; P = 0.25), though neither finding attained statistical significance. CONCLUSIONS: This exploratory study was underpowered to detect differences between the groups. Further studies are needed to reliably compare pain and ecchymosis in LMWH vs. UFH. PMID- 19219406 TI - Simultaneous visualization of two protein complexes in a single plant cell using multicolor fluorescence complementation analysis. AB - Bimolecular fluorescence complementation (BiFC) is an approach used to analyze protein-protein interaction in vivo, in which non-fluorescent N-terminal and C terminal fragments of a fluorescent protein are reconstituted to emit fluorescence only when they are brought together by interaction of two proteins to fuse both fragments. A method for simultaneous visualization of two protein complexes by multicolor BiFC with fragments from green fluorescent protein (GFP) and its variants such as cyan and yellow fluorescent proteins (CFP and YFP) was recently reported in animal cells. In this paper we describe a new strategy for simultaneous visualization of two protein complexes in plant cells using the multicolor BiFC with fragments from CFP, GFP, YFP and a red fluorescent protein variant (DsRed-Monomer). We identified nine different BiFC complexes using fragments of CFP, GFP and YFP, and one BiFC complex using fragments of DsRed Monomer. Fluorescence complementation did not occur by combinations between fragments of GFP variants and DsRed-Monomer. Based on these findings, we achieved simultaneous visualization of two protein complexes in a single plant cell using two colored fluorescent complementation pairs (cyan/red, green/red or yellow/red). PMID- 19219407 TI - Acute myocardial infarction following oral methyl-ergometrine intake. AB - Ergot derivatives are frequently administrated during cesarean delivery, induced abortion, or post-partum hemorrhage to promote uterine contractions. Ergot derivatives may also induce coronary spasm and intravenous ergonovine is used in cardiac catheterization laboratories as a diagnostic agent. Serious ischemic cardiac events related to ergonovine are rare and have most often been described after intravenous use. We report the case of a 38-year-old woman with a ST elevation myocardial infarction (STEMI) few days after artificially induced abortion by oral prescription of methylergometrine. Coronary angiography performed 2 days after onset of chest pain did not reveal any abnormalities of the coronary arteries but a provocative test using intravenous methylergometrine was positive with reproduction of chest pain, ECG changes and with a significant narrowing localized on the second segment of the left anterior descending artery at the angiogram. Thus, since methylergometrin may clearly induce coronary spasm when prescribed orally, chest pain occurring under oral treatment should require immediate attention even if atypical or in the absence of cardiovascular risk factors. PMID- 19219408 TI - Pharmacokinetics of digoxin in healthy subjects receiving taranabant, a novel cannabinoid-1 receptor inverse agonist. AB - INTRODUCTION: Interaction studies with digoxin (Lanoxin; GlaxoSmithKline, Research Triangle Park, NC, USA), a commonly prescribed cardiac glycoside with a narrow therapeutic index and a long half-life, are typically required during the development of a new drug, particularly when it is likely that digoxin may be given to patients also treated with the new agent, taranabant--a cannabinoid-1 receptor inverse agonist--for weight loss. This study was designed to establish if this combination of therapy has the potential of a significant pharmacokinetic interaction. METHODS: This open-label, fixed-sequence, two-period study investigated whether taranabant, administered to steady state, affects the well described single-dose pharmacokinetics of digoxin. During the first period, 12 healthy men and women ranging in age from 21 to 35 years received a single oral dose of digoxin 0.5 mg. Following a 10-day wash out, they started a 19-day taranabant dosing regimen (6 mg once daily from day -14 to day 5) designed to establish and maintain steady-state levels of taranabant. On study day 1, subjects received a single oral dose of digoxin 0.5 mg. The plasma levels of digoxin were followed for an additional 4 days while the dosing of taranabant continued. RESULTS: The geometric mean ratio and 90% confidence intervals for digoxin AUC(0-infinity) were 0.91 (0.83, 0.99), falling within the prespecified comparability intervals (CI) of (0.8, 1.25), which is within the usually allowed interval for bioequivalence. The geometric mean ratio and 90% CI for digoxin maximum plasma concentration (C(max)) were 1.23 (1.09, 1.40). The median time to C(max) was the same for both treatments. CONCLUSION: Multiple doses of 6 mg taranabant do not have a clinically meaningful effect on the pharmacokinetics of a single oral dose of digoxin. PMID- 19219409 TI - Comparison of costs among patients with type 2 diabetes treated with exenatide or sitagliptin therapy. AB - INTRODUCTION: Exenatide (Byetta, Amylin Pharmaceuticals Inc., CA, USA) and sitagliptin (Januvia, Merck & Co, NJ, USA) are two antidiabetic agents recently approved by the US Food and Drug Administration. The purpose of this analysis was to compare costs among patients with type 2 diabetes (T2D) treated with either of these agents. METHODS: Data with dates of service from September 1, 2005 through August 31, 2007, were obtained from a large US retrospective claims database. Intent-to-treat cohorts of adults diagnosed with T2D who began taking either exenatide (n=1885) or sitagliptin (n=2482) and did not use the alternate medication in the 6-month follow-up period were created. Six-month total medical costs were estimated using stepwise multivariate regressions. Six-month total diabetes-related medical costs, a component of total medical costs, were also estimated using stepwise multivariate regressions. In addition, other cost components were examined using either stepwise multivariate regressions or a two part model that controlled for the probability of using the medical service. Smearing estimates were used to transform estimated log costs into costs. The analysis controlled for the potential impact of patient demographics, general health, prior resource use, comorbidities, and timing of treatment initiation. RESULTS: Exenatide was associated with lower total 6-month direct medical costs ($9340 vs. $9995; P<0.0001), despite some component costs being slightly higher with exenatide: diabetes-related drug costs ($1765 vs. $1743; P=0.0062), diabetes related medical costs ($4142 vs. $4002; P<0.0001), and emergency room costs ($43 vs. $29; P=0.0388). Exenatide was associated with lower outpatient costs ($4498 vs. $5942; P<0.0001). CONCLUSIONS: Compared with the use of sitagliptin, exenatide was associated with lower total medical costs (difference of $655) despite higher total diabetes-related costs (difference of $140). As a result, there appears to be overall cost savings associated with the use of exenatide relative to sitagliptin. PMID- 19219410 TI - Cost-effectiveness analysis of lopinavir/ritonavir and atazanavir+ritonavir regimens in the CASTLE study. AB - INTRODUCTION: The purpose of the study was to conduct a cost-effectiveness analysis and budget impact analysis comparing lopinavir with ritonavir (LPV/r) and atazanavir plus ritonavir (ATV+RTV) for antiretroviral-naive patients with a baseline CD4+ T-cell distribution and total cholesterol (TC) profile as reported in the CASTLE study. METHODS: This decision analysis study used a previously published Markov model of HIV disease, incorporating coronary heart disease (CHD) events to compare the short- and long-term budget impacts and CHD consequences expected for the two regimens. RESULTS: Patients were assumed to have a baseline CHD risk of 4.6% (based on demographic data) and it was also assumed that 50% of the population in the CASTLE study were smokers. The CHD risk differences (based on percent of patients with TC >240 mg/dL) in favor of ATV+RTV resulted in an average improvement in life expectancy of 0.031 quality-adjusted life years (QALYs) (11 days), and an incremental cost-effectiveness ratio of $1,409,734/QALY. Use of the LPV/r regimen saved $24,518 and $36,651 at 5 and 10 years, respectively, with lifetime cost savings estimated at $38,490. A sensitivity analysis using a cohort of all smokers on antihypertensive medication estimated an average improvement in life expectancy of 31 quality-adjusted days in favor of ATV+RTV, and a cost-effectiveness ratio of $520,861/QALY: a ratio that is still above the acceptable limit within the US. CONCLUSION: The use of an LPV/r-based regimen in antiretroviral-naive patients with a baseline CHD risk similar to patients in the CASTLE study appears to be a more cost-effective use of resources compared with an ATV+RTV-based regimen. The very small added CHD risk predicted by LPV/r treatment is more than offset by the substantial short- and long-term cost savings expected with the use of LPV/r in antiretroviral-naive individuals with average to moderately elevated CHD risk. PMID- 19219411 TI - Qualify of life and palliation predict survival in patients with chronic alveolar hypoventilation and nocturnal ventilatory support. AB - OBJECTIVES: Non-invasive positive pressure ventilation (NPPV) improves health related quality of life (HRQL) in patients with chronic alveolar hypoventilation (CAH). We studied the prognostic impact of HRQL on survival in relation to clinical factors. PATIENTS: Forty-four patients with CAH due to post-polio (12), scoliosis (11), post-tb (17) or other diagnoses (4) who received nocturnal NPPV were prospectively studied during 6-10 years. MEASUREMENTS: Blood gases and HRQL were analysed at baseline and after 9 months and after 8 years. HRQL was evaluated with measures of functioning (SIP), emotional well-being (HADS and MACL), and global QL. RESULTS: Blood gases and HRQL measures improved during NPPV. The overall 5-year survival rate was 73%. In multivariate survival analysis, a diagnosis of post-polio and low baseline SIP physical index scores, indicating low levels of physical dysfunction, predicted longer survival (P = 0.02, respectively). Similarly, palliation of physical dysfunction and preserved or improved global QL by 9 months were associated with longer overall survival (P = 0.009 and P = 0.001, respectively; multivariate Cox regression). CONCLUSION: Seventy-three percent of patients treated for CAH with NPPV survived more than 5 years. Diagnosis and self-rated physical functioning at pre-treatment were related to survival, as were major improvements in physical functioning and global QL during NPPV. PMID- 19219412 TI - [Perioperative management of patients with obstructive sleep apnoea]. AB - There is lack of studies investigating procedures aiming at a decrease in perioperative mortality in patients with obstructive sleep apnoea (OSA). During anesthetic evaluation, identification of patients with OSA as well as using a risk score has been recommended by the American Society of Anesthesiology in order to identify the best perioperative strategy. Perioperative attention should be focused on a secure airway and the duration of monitoring that is necessary regarding severity of OSA, surgical stress and respiratory function. Postoperatively, residual neuromuscular blockade and a supine position have to be avoided. Continuous pulse oximetry should be used as long as patients remain at increased risk and should be applied until oxygen saturation remains above 90% with room air during sleep. Opioids should be excluded for pain management whenever possible, and CPAP or NIPPV should be administered as soon as feasible after surgery to patients who have been receiving it preoperatively. PMID- 19219413 TI - [Prehospital obstetrical emergencies in a physician-staffed ground-based emergency service. A retrospective analysis over a 5-year period]. AB - BACKGROUND: In the German emergency medical system (EMS) obstetrical emergencies are rarely encountered, but are highly emotional situations for all concerned and form a special challenge for the emergency physician. The aim of this study was to evaluate the incidence, the course and the performance of rescue missions in a ground-based EMS system. METHODS: In a retrospective study the prehospital emergency charts concerning obstetrical emergencies over a 5-year period (10/2002 09/2007) were analysed. RESULTS: A total of 40 physician-staffed rescue missions with obstetrical emergencies were identified. On average seven rescue missions were performed per year. The majority of cases with 73% of the rescue missions was performed during the night service (16:00-07:00 h). On average the emergency patients (26th-41st week of gestation) were classified by the National Advisory Committee for Aeronautics (NACA) score as NACA III. Of the 40 obstetrical emergencies delivery occurred out of hospital in 18 cases (33rd-41st week of gestation), while the emergency physician was present in only 3 cases during childbirth. In 15 cases prehospital childbirth took place in the domestic environment of the patient, in 2 cases in an ambulance and in 1 case in the medical office of a gynecologist. In 20 cases the pregnant women were transported to hospital while labor had already begun. The emergency physicians on scene applied intravenous access, guided through labor and delivery, and administered tocolysis and in cases of prehospital delivery the emergency physicians also applied oxytocin, cut the umbilical cord and performed primary care of the newborn. CONCLUSIONS: Obstetrical emergencies are rare but recurrent in the ground-based EMS. However, prehospital management of women in labor, supervision of spontaneous prehospital delivery and the initial management of a newborn form a challenge for the emergency physician responsible. Consequently, prehospital management of obstetrical emergencies needs intensive consideration during education and training of emergency medical personnel. PMID- 19219415 TI - [Technical aspects of laparoscopic Heller myotomy for achalasia]. AB - Minimally invasive surgery has influenced the treatment of achalasia more than that of any other gastrointestinal disorder. Laparoscopic Heller myotomy has thus evolved to the first-line therapy in patients with achalasia and led to a significant change in the treatment algorithm of this disorder. The aim of this article is to present technical aspects and pitfalls of Heller myotomy with combined antirefluxplasty. After injection of 0.9% NaCl into the muscularis and submucosa of the distal esophagus and proximal fundus, whereby the submucosal layer can be easily separated from the mucosa, myotomy of the longitudinal and circular musculature is performed up to 6-7 cm proximally to the esophagogastric junction and completed distally by 1.5-2 cm onto the fundus. We prefer the 180 degrees anterior semifundoplication according to Dor as antirefluxplasty, which is sutured in a two-rowed manner into the two sites of the myotomy. The pitfalls are incomplete myotomy, especially at its distal, fundic site, with consecutive persistence or recurrence of symptoms, as well as occult mucosal perforations, which can be detected by intraoperative endoscopy. PMID- 19219416 TI - [Relevance of ESBL-producing pathogens for clinical surgery: diagnostics, therapy, and prevention]. AB - BACKGROUND: Multiresistant, extended spectrum beta lactamase (ESBL)-producing pathogens are an increasing problem in daily clinical life. This paper summarizes the development of resistance as well as epidemiology, diagnostics, and treatment of ESBL-producing micro-organisms. We analyzed microbiological data collected at the Grosshadern Clinic in Germany between 1996 and 2007, in order to assess the importance of these micro-organisms to medical practice and surgical care units. PATIENTS AND METHODS: Pathogens were isolated from 28,894 patients with Escherichia coli and 10,903 with Klebsiella pneumoniae pathogens between 1996 and 2006 and tested for ESBL production. For the year 2007 we have analyzed the complete spectrum of ESBL-producing pathogens and their distribution to different departments of the clinic. The agar diffusion test with five cephalosporins and an automated detection system (BD Phoenix) were used for screening purposes. Positive results were verified with the E- and double-disc agar diffusion tests. RESULTS: The most important pathogens isolated from patients were E. coli and K. pneumoniae. Analysis of ESBL-producing E. coli pathogens from 1996 to 2006 showed the prevalence increasing from 0% to 4.1%. For ESBL-producing K. pneumoniae, we also found a prevalence rising from 0.3% in 1996 to 6.6% in 2006. For the year 2007 a further increase in ESBL-producing pathogens was detected, reaching 182 cases, with 118 of ESBL-producing E. coli (5.7 %) and 39 of ESBL-producing K. pneumoniae (7.4%). Of these, 24 cases with E. coli and nine with K. pneumoniae were surgery patients (20% and 23%, respectively). CONCLUSION: The results show an increasing prevalence of ESBL-producing pathogens in hospitalized patients and in surgical departments. The resulting rise in treatment costs and patient risk require thorough knowledge of risk factors, therapy, and preventive measures. PMID- 19219417 TI - [31-year-old male patient with testicular mass and hyperthyroidism]. AB - An excess of human chorionic gonadotropin (hCG) is a rare differential diagnosis of hyperthyroidism, due to the TSH-like effect of hCG. hCG levels physiologically increase during pregnancy. In patients with germ cell tumors of the testicles and the ovary, a pathological increase of hCG serum concentrations may also be found, depending on the histological subtype. We report on a 31-year-old male patient with overt clinical hyperthyroidism resulting of a hCG-producing metastasized germ cell tumor of the right testicle. After initiation of chemotherapy, thyroid function normalized in parallel to the tumor's remission with marked decrease of hCG levels. PMID- 19219418 TI - IL-22: a critical mediator in mucosal host defense. AB - IL-22 is an IL-10 family cytokine member that was recently discovered to be produced by Th17 cells. Current studies have revealed that IL-22 targets cells of the digestive, skin, and respiratory organ systems and plays an important role in mucosal immunity. The IL-22 receptor (IL-22R) is expressed exclusively in these tissues, thereby allowing the cytokine to mediate epithelial innate immunity in response to a variety of pathogens. Although recent studies have shown the importance of IL-22 in host defense against Gram-negative bacterial organisms (in gut and lung), there is evidence that IL-22 also plays a role in autoimmune disease, such as psoriasis. IL-22 therefore, not unlike other cytokines, has complex pro-inflammatory, anti-inflammatory, and autoimmune effects which continue to be under further investigation. This review will focus on what is known about IL-22 and its function in mucosal host defense. PMID- 19219419 TI - Human antibodies targeting cell surface antigens overexpressed by the hormone refractory metastatic prostate cancer cells: ICAM-1 is a tumor antigen that mediates prostate cancer cell invasion. AB - Transition from hormone-sensitive to hormone-refractory metastatic tumor types poses a major challenge for prostate cancer treatment. Tumor antigens that are differentially expressed during this transition are likely to play important roles in imparting prostate cancer cells with the ability to grow in a hormone deprived environment and to metastasize to distal sites such as the bone and thus, are likely targets for therapeutic intervention. To identify those molecules and particularly cell surface antigens that accompany this transition, we studied the changes in cell surface antigenic profiles between a hormone sensitive prostate cancer line LNCaP and its hormone-refractory derivative C4-2B, using an antibody library-based affinity proteomic approach. We selected a naive phage antibody display library to identify human single-chain antibodies that bind specifically to C4-2B but not LNCaP. Using mass spectrometry, we identified one of the antibody-targeted antigens as the ICAM-1/CD54/human rhinovirus receptor. Recombinant IgG1 derived from this single-chain antibody binds to a neutralizing epitope of ICAM-1 and blocks C4-2B cell invasion through extracellular matrix in vitro. ICAM-1 is thus differentially expressed during the transition of the hormone-sensitive prostate cancer cell line LNCaP to its hormone-refractory derivative C4-2B, plays an important role in imparting the C4 2B line with the ability to invade, and may therefore be a target for therapeutic intervention. PMID- 19219421 TI - Potential reno-protective effects of a gluten-free diet in type 1 diabetes. AB - AIMS/HYPOTHESIS: Coeliac disease is common in type 1 diabetes. It is managed with a gluten-free diet, characterised by foods low in AGEs. We hypothesised that this diet would lead to lower plasma AGEs and be associated with reduced albuminuria. METHODS: From a single paediatric clinic, we recruited 21 children with type 1 diabetes and biopsy-proven coeliac disease, and 38 individuals with diabetes alone. The groups were matched for age, sex, duration of disease and metabolic control. Participants completed a detailed clinical and dietary history. Blood samples were taken for HbA(1c), coeliac serology, thyroid function, serum IgA levels and plasma AGEs, and urine samples were obtained for estimation of the albumin/creatinine ratio (ACR). RESULTS: All the individuals with coeliac disease were asymptomatic, with negative transglutaminase antibodies. There were no significant differences between the groups in terms of age (14 years), sex (29% male), duration of diabetes (7 years), mean HbA(1c) (8.3%), lipid levels or treatment regimens. However, children with diabetes and coeliac disease had twofold lower levels of urinary ACR than with those diabetes alone (p = 0.04). This was associated with lower levels of circulating AGEs (p = 0.03). These associations were independent of metabolic control, diabetes management and other potentially confounding variables, such as household exposure to cigarette smoke. CONCLUSIONS/INTERPRETATION: Adherence to a gluten-free diet may provide additional benefits for individuals with coeliac disease, and potentially those with type 1 diabetes. PMID- 19219420 TI - Adults with type 1 diabetes eat a high-fat atherogenic diet that is associated with coronary artery calcium. AB - AIMS/HYPOTHESIS: Coronary heart disease is the leading cause of mortality among people with type 1 diabetes. Diet is an important lifestyle factor that relates to risk of CHD. The aim of this study was to examine how diet and adherence to dietary guidelines differ between adults with and without type 1 diabetes, and their correlation with CHD risk factors and coronary artery calcium (CAC). METHODS: The study involved 571 people with type 1 diabetes and 696 controls, aged 19 to 56 years, who were asymptomatic for CHD. CAC was measured by electron beam computed tomography. RESULTS: Compared with the controls, adults with type 1 diabetes reported a diet higher in fat, saturated fat and protein but lower in carbohydrates. Fewer than half of those with type 1 diabetes met dietary guidelines for fat and carbohydrate intake, and only 16% restricted saturated fat to less than 10% of daily energy intake. Adults with type 1 diabetes were significantly less likely to meet dietary guidelines than controls. Fat and saturated fat intakes were positively correlated, but carbohydrate intake was negatively correlated with CHD risk factors and HbA(1c). A high-fat diet and higher intake of protein were associated with greater odds of CAC, while higher carbohydrate intake was associated with reduced odds of CAC. CONCLUSIONS/INTERPRETATION: Adults with type 1 diabetes reported consuming higher than recommended levels of fat and saturated fat. High fat intake was associated with increased CHD risk factors, worse glycaemic control and CAC. An atherogenic diet may contribute to the risk of CHD in adults with type 1 diabetes. PMID- 19219422 TI - Involvement of TNF-alpha in abnormal adipocyte and muscle sortilin expression in obese mice and humans. AB - AIMS/HYPOTHESIS: Insulin resistance is caused by numerous factors including inflammation. It is characterised by defective insulin stimulation of adipocyte and muscle glucose transport, which requires the glucose transporter GLUT4 translocation towards the plasma membrane. Defects in insulin signalling can cause insulin resistance, but alterations in GLUT4 trafficking could also play a role. Our goal was to determine whether proteins controlling GLUT4 trafficking are altered in insulin resistance linked to obesity. METHODS: Using real-time RT PCR, we searched for selected transcripts that were differentially expressed in adipose tissue and muscle in obese mice and humans. Using various adipocyte culture models and in vivo mice treatment, we searched for the involvement of TNF alpha in these alterations in obesity. RESULTS: Sortilin mRNA and protein were downregulated in adipose tissue from obese db/db and ob/ob mice, and also in muscle. Importantly, sortilin mRNA was also decreased in morbidly obese human diabetic patients. Sortilin and TNF-alpha (also known as TNF) mRNA levels were inversely correlated in mice and human adipose tissues. TNF-alpha decreased sortilin mRNA and protein levels in cultured mouse and human adipocytes, an effect partly prevented by the peroxisome proliferator-activated receptor gamma activator rosiglitazone. TNF-alpha also inhibited adipocyte and muscle sortilin mRNA when injected to mice. CONCLUSIONS/INTERPRETATION: Sortilin, an essential player in adipocyte and muscle glucose metabolism through the control of GLUT4 localisation, is downregulated in obesity and TNF-alpha is likely to be involved in this defect. Chronic low-grade inflammation in obesity could thus contribute to insulin resistance by modulating proteins that control GLUT4 trafficking. PMID- 19219424 TI - Optical diagnosis of peritoneal metastases by infrared microscopic imaging. AB - Fourier transform infrared (FTIR) spectroscopy is nowadays widely accepted as a technique with high potential for diagnosis of cancerous tissues. This study presents an example of the investigation of peritoneal metastases by FTIR microimaging. Peritoneal malignancies are generally secondary localizations of primary visceral cancers such as ovarian, stomach or colon cancers. By analysing simultaneously both formalin-fixed paraffin-embedded and frozen specimens, we examined malignant and non-malignant (i.e. fibrotic and cicatricial) peritoneal lesions. Paraffin-embedded tissues were analysed without any previous dewaxing. Multivariate statistical approaches, based on the classification of infrared data by hierarchical cluster analysis, allowed the discrimination of these various samples. Microimaging also permits the revelation of the heterogeneity of the tissue: it was possible to localize precisely the cancerous areas, and to distinguish, on the basis of their spectral signatures, the peritumoral neighbouring connective tissue close to the carcinomatous areas from the connective tissue distant from the cancerous areas. These spectral differences could be useful as complementary information to study molecular changes associated with the malignancy. PMID- 19219423 TI - Endothelial inflammation induced by excess glucose is associated with cytosolic glucose 6-phosphate but not increased mitochondrial respiration. AB - AIMS/HYPOTHESIS: Exposure of endothelial cells to high glucose levels suppresses responses to insulin, including induction of endothelial nitric oxide synthase activity, through pro-inflammatory signalling via the inhibitor of nuclear factor kappaB (IkappaB)alpha-nuclear factor kappaB (NF-kappaB) pathway. In the current study, we aimed to identify metabolic responses to glucose excess that mediate endothelial cell inflammation and insulin resistance. Since endothelial cells decrease their oxygen consumption rate (OCR) in response to glucose, we hypothesised that increased mitochondrial function would not mediate these cells' response to excess substrate. METHODS: The effects of glycolytic and mitochondrial fuels on metabolic intermediates and end-products of glycolytic and oxidative metabolism, including glucose 6-phosphate (G6P), lactate, CO(2), NAD(P)H and OCR, were measured in cultured human microvascular endothelial cells and correlated with IkappaBalpha phosphorylation. RESULTS: In response to increases in glucose concentration from low to physiological levels (0-5 mmol/l), production of G6P, lactate, NAD(P)H and CO(2) each increased as expected, while OCR was sharply reduced. IkappaBalpha activation was detected at glucose concentrations >5 mmol/l, which was associated with parallel increases of G6P levels, whereas downstream metabolic pathways were insensitive to excess substrate. CONCLUSIONS/INTERPRETATION: Phosphorylation of IkappaBalpha by excess glucose correlates with increased levels of the glycolytic intermediate G6P, but not with lactate generation or OCR, which are inhibited well below saturation levels at physiological glucose concentrations. These findings suggest that oxidative stress due to increased mitochondrial respiration is unlikely to mediate endothelial inflammation induced by excess glucose and suggests instead the involvement of G6P accumulation in the adverse effects of hyperglycaemia on endothelial cells. PMID- 19219426 TI - Potential roles of force cues in human stance control. AB - Human stance is inherently unstable. A small deviation from upright body orientation is enough to yield a gravitational component in the ankle joint torque, which tends to accelerate the body further away from upright ('gravitational torque'; magnitude is related to body-space lean angle). Therefore, to maintain a given body lean position, a corresponding compensatory torque must be generated. It is well known that subjects use kinematic sensory information on body-space lean from the vestibular system for this purpose. Less is known about kinetic cues from force/torque receptors. Previous work indicated that they are involved in compensating external contact forces such as a pull or push having impact on the body. In this study, we hypothesized that they play, in addition, a role when the vestibular estimate of the gravitational torque becomes erroneous. Reasons may be sudden changes in body mass, for instance by a load, or an impairment of the vestibular system. To test this hypothesis, we mimicked load effects on the gravitational torque in normal subjects and in patients with chronic bilateral vestibular loss (VL) with eyes closed. We added/subtracted extra torque to the gravitational torque by applying an external contact force (via cable winches and a body harness). The extra torque was referenced to body space lean, using different proportionality factors. We investigated how it affected body-space lean responses that we evoked using sinusoidal tilts of the support surface (motion platform) with different amplitudes and frequencies (normals +/-1 degrees, +/-2 degrees, and +/-4 degrees at 0.05, 0.1, 0.2, and 0.4 Hz; patients +/-1 degrees and +/-2 degrees at 0.05 and 0.1 Hz). We found that added/subtracted extra torque scales the lean response in a systematic way, leading to increase/decrease in lean excursion. Expressing the responses in terms of gain and phase curves, we compared the experimental findings to predictions obtained from a recently published sensory feedback model. For the trials in which the extra torque tended to endanger stance control, predictions in normals were better when the model included force cues than without these cues. This supports our notion that force cues provide an automatic 'gravitational load compensation' upon changes in body mass in normals. The findings in the patients support our notion that the presumed force cue mechanism provides furthermore vestibular loss compensation. Patients showed a body-space stabilization that cannot be explained by ankle angle proprioception, but must involve graviception, most likely by force cues. Our findings suggest that force cues contribute considerably to the redundancy and robustness of the human stance control system. PMID- 19219427 TI - To what extent does the indicator "concurrent use of three or more psychotropic drugs" capture use of potentially inappropriate psychotropics among the elderly? AB - PURPOSE: The indicator "concurrent use of three or more psychotropic drugs" has been used as a measure of quality in drug use among the elderly. The aim of our study was to assess to what extent the indicator captures the use of specific psychotropics associated with an increased risk of adverse events among the elderly, i.e., potentially inappropriate psychotropic drugs (PIP). METHODS: All individuals aged 75 years and older in Sweden purchasing prescribed psychotropic drugs in 2006 constituted the study population (n = 384,904). Data on purchased psychotropic drugs from the Swedish Prescribed Drug Register were used. The overlap between individuals with the indicator and individuals using PIP was assessed with sensitivity, specificity, positive and negative predictive values and likelihood ratio as outcome measures. RESULTS: Among the psychotropic drug users, 15% had the indicator and 39% used PIP. The proportion of individuals with the indicator among all individuals using PIP was 27% (sensitivity). The proportion of individuals without the indicator among all individuals not using PIP was 93% (specificity). The positive predictive value was 72%, and the negative predictive value was 67%. Differences in outcome measures were observed between different categories of PIP. CONCLUSIONS: The indicator "concurrent use of three or more psychotropics" can be technically easy to use, but PIP is more specific. Three quarters of all individuals who used PIP in this study were not captured by the indicator. However, two thirds of all individuals with the indicator used PIP. When selecting instruments to assess appropriateness in drug therapy in the elderly, clinical relevance should be balanced against convenience of use. PMID- 19219428 TI - Paraffin oil as a "methane vector" for rapid and high cell density cultivation of Methylosinus trichosporium OB3b. AB - Slow growth and relatively low cell densities of methanotrophs have limited their uses in industrial applications. In this study, a novel method for rapid cultivation of Methylosinus trichosporium OB3b was studied by adding a water immiscible organic solvent in the medium. Paraffin oil was the most effective at enhancing cell growth and final cell density. This is at least partially due to the increase of methane gas transfer between gas and medium phases since methane solubility is higher in paraffin than in water/nitrate minimal salt medium. During cultivation with paraffin oil at 5% (v/v) in the medium, M. trichosporium OB3b cells also showed higher concentrations of the intermediary metabolites, such as formic acid and pyruvic acid, and consumed more methane compared with the control. Paraffin as methane vector to improve methanotroph growth was further studied in a 5-L fermentor at three concentrations (i.e., 2.5%, 5%, and 10%). Cell density reached about 14 g dry weight per liter with 5% paraffin, around seven times higher than that of the control (without paraffin). Cells cultivated with paraffin tended to accumulate around the interface between oil droplets and the water phase and could exist in oil phase in the case of 10% (v/v) paraffin. These results indicated that paraffin could enhance methanotroph growth, which is potentially useful in cultivation of methanotrophs in large scale in industry. PMID- 19219429 TI - (18)F-FDG PET for the evaluation of thymic epithelial tumors: Correlation with the World Health Organization classification in addition to dual-time-point imaging. AB - PURPOSE: Our aim was to determine dual-phase (18)F-FDG PET imaging features for various subtypes of thymic epithelial tumors based on the World Health Organization classification. METHODS: Forty-six patients with histologically verified thymic epithelial tumors [23 with low-risk tumors (4 with type A, 16 with AB, and 3 with B1) and 23 with high-risk tumors (7 with B2, 5 with B3, and 11 with thymic carcinoma] were enrolled in this study. All patients were injected with (18)F-FDG.; after 1 h, they underwent scanning; after 3 h, 23 patients underwent an additional scanning. The maximum standard uptake value (SUV(max)) and the retention index (RI%) of the lesions were determined. RESULTS: The early and delayed SUV(max) values in the patients with high-risk tumors [early SUV(max) (mean: 6.0) and delayed SUV(max) (mean: 7.4)] were both significantly larger than those in patients with low-risk tumors [early SUV(max) (mean: 3.2) and delayed SUV(max) (mean: 3.4)] (P < 0.05). Early SUV(max) values of greater than 7.1 differentiated thymic carcinomas from other types of tumors. For the histological differentiation between high-risk tumors and low-risk tumors, an early SUV(max) value of 4.5 was used as the cutoff. The sensitivity, specificity, and accuracy were 78.3, 91.3, and 84.8%, respectively. CONCLUSION: High SUV values (early SUV > 4.5) suggest the presence of high-risk tumors. A very high SUV value (early SUV > 7.1) is useful for the differentiation of thymic carcinomas from other types of tumors. The delayed SUV values were higher than the early SUV values in all types of tumors. PMID- 19219430 TI - Association between FDG uptake, CSF biomarkers and cognitive performance in patients with probable Alzheimer's disease. AB - PURPOSE: Brain imaging of FDG uptake and cerebrospinal fluid (CSF) concentration of amyloid-beta 1-42 (Abeta(1-42)) or tau proteins are promising biomarkers in the diagnosis of Alzheimer's disease (AD). There is still uncertainty regarding any association between decreased FDG uptake and alterations in CSF markers. METHODS: The relationship between FDG uptake, CSF Abeta(1-42) and total tau (T tau), as well as the Mini-Mental State Examination (MMSE) score was investigated in 34 subjects with probable AD using step-wise linear regression. FDG uptake was scaled to the pons. RESULTS: Scaled FDG uptake was significantly reduced in the probable AD subjects compared to 17 controls bilaterally in the precuneus/posterior cingulate area, angular gyrus/inferior parietal cortex, inferior temporal/midtemporal cortex, midfrontal cortex, and left caudate. Voxel based single-subject analysis of the probable AD subjects at p < 0.001 (uncorrected) revealed a total volume of significant hypometabolism ranging from 0 to 452 ml (median 70 ml). The total hypometabolic volume was negatively correlated with the MMSE score, but it was not correlated with the CSF measures. VOI-based step-wise linear regression revealed that scaled FDG uptake in the precuneus/posterior cingulate was negatively correlated with CSF Abeta(1-42). Scaled FDG uptake in the caudate was positively correlated with CSF T-tau. CONCLUSION: The extent and local severity of the reduction in FDG uptake in probable AD subjects are associated with cognitive impairment. In addition, there appears to be a relationship between local FDG uptake and CSF biomarkers which differs between different brain regions. PMID- 19219431 TI - Improved quantification in single-pinhole and multiple-pinhole SPECT using micro CT information. AB - PURPOSE: The purpose of this study was to demonstrate the feasibility of accurate quantification in pinhole SPECT using micro-CT information. METHODS: Pinhole SPECT scans were performed using a clinical dual-head gamma camera. Each pinhole SPECT scan was followed by a micro-CT acquisition. Functional and anatomical images were coregistered using six point sources visible with both modalities. Pinhole SPECT images were reconstructed iteratively. Attenuation correction was based on micro-CT information. Scatter correction was based on dual and triple energy window methods. Phantom and animal experiments were performed. A phantom containing nine vials was filled with different concentrations of (99m)Tc. Three vials were also filled with CT contrast agent to increase attenuation. Activity concentrations measured on the pinhole SPECT images were compared with activity concentrations measured by the dose calibrator. In addition, 11 mice were injected with (99m)Tc-labelled Nanobodies. After acquiring functional and anatomical images, the animals were killed and the liver activity was measured using a gamma-counter. Activity concentrations measured on the reconstructed images were compared with activity concentrations measured with the gamma counter. RESULTS: The phantom experiments demonstrated an average error of -27.3 +/- 15.9% between the activity concentrations measured on the uncorrected pinhole SPECT images and in the dose calibrator. This error decreased significantly to 0.1 +/- 7.3% when corrections were applied for nonuniform attenuation and scatter. The animal experiment revealed an average error of -18.4 +/- 11.9% between the activity concentrations measured on the uncorrected pinhole SPECT images and measured with the gamma counter. This error decreased to -7.9 +/- 10.4% when attenuation and scatter correction was applied. CONCLUSION: Attenuation correction obtained from micro-CT data in combination with scatter correction allows accurate quantification in pinhole SPECT. PMID- 19219432 TI - Pulsed lavage reduces the incidence of radiolucent lines under the tibial tray of Oxford unicompartmental knee arthroplasty: pulsed lavage versus syringe lavage. AB - The aim of the study was to determine whether the incidence of radiolucencies can be reduced using pulsed lavage before cementing the tibia in unicompartmental knee arthroplasty (UKA). We prospectively studied a consecutive series of 112 cemented Oxford UKA in 100 patients in two centres. In group A (n = 56) pulsed lavage and in group B (n = 56) conventional syringe lavage was used to clean the cancellous bone. The same standardised cementing technique was applied in all cases. At a minimum follow-up of one year patients were evaluated clinically and screened radiographs were obtained. The cement bone interface under the tibial plateau was divided into four zones and evaluated for the presence of radiolucent lines. All radiographs were evaluated (n = 112), and radiolucencies in all four zones were found in two cases in group A (4%) and in 12 cases in group B (22%) (p = 0.0149). Cement penetration showed a median of 2.6 mm (group A) and 1.5 mm (group B) (p < 0.0001). We recommend the routine use of pulsed lavage in Oxford UKA to reduce the incidence of radiolucency and to improve long-term fixation. PMID- 19219433 TI - Association of oestrogen receptor gene polymorphism with the long-term results of rotational acetabular osteotomy. AB - Acetabular dysplasia (AD) contributes to the development of osteoarthritis of the hip. A rotational acetabular osteotomy (RAO) is one of the methods of pelvic osteotomy to prevent or treat secondary osteoarthritis of the hip. Although most of the patients that undergo RAO show satisfactory results, some have poor results. This study investigated whether gene polymorphisms of both the vitamin D receptor (VDR) and oestrogen receptor (ER) are involved in both AD and the postoperative results following RAOs. Sixty-four Japanese patients with AD who were treated by an RAO were enrolled in this study (59 women and 5 men, aged 13 59, with an average age of 40.3). Gene polymorphisms of the VDR [ApaI and TaqI restriction fragment length polymorphisms (RFLPs)] and ER (PvuII and XbaI RFLPs) were determined in these patients. The relationship between both the AD and radiographic postoperative changes of the hip joint after an RAO with these gene polymorphisms were examined. The frequencies of ER gene polymorphism coded as pp (RFLP/PvuII) in patients with AD were statistically significantly different (p = .011) from those coded as both PP and Pp. The joint space width narrowed even after RAO in 90% of the patients with the pp gene polymorphism, while it narrowed in only 35% of the patients with either PP or Pp seven years or longer after an RAO. The PvuII polymorphism in the ER gene was associated with the postoperative result of an RAO, while no association was observed between the AD with VDR and ER gene polymorphisms. PMID- 19219434 TI - Metal release and corrosion effects of modular neck total hip arthroplasty. AB - Modular neck implants are an attractive treatment tool in total hip replacement. Concerns remain about the mechanical stability and metal ion release caused by the modular connection. Five different implant designs were investigated in an experimental set-up. In vivo conditions were simulated and the long-term titanium release was measured. Finally, the modular connections were inspected for corrosion processes and signs of fretting. No mechanical failure or excessive corrosion could be identified for the implants tested. The titanium releases measured were extremely low compared to in vivo and in vitro studies and were not in a critical range. PMID- 19219436 TI - [Nobel Prize in medicine]. PMID- 19219435 TI - [Genome-wide expression profiling as a clinical tool: are we there yet?]. AB - Breast cancer is a heterogeneous disease, encompassing a plethora of histological types and clinical courses. Current histopathological classification systems for breast cancer are based on descriptive entities that are of prognostic significance. Few prognostic markers beyond those offered by histopathological analysis are available. Furthermore, a very limited armamentarium of predictive biomarkers has been introduced in clinical practice. High throughput molecular technologies are reshaping our understanding of breast cancer, of which microarray-based gene expression has received the most attention. This method has been successfully used to derive a molecular taxonomy for breast cancer, which has provided interesting insights into the biology of the disease. Microarray based class prediction studies have generated a multitude of prognostic/predictive signatures. Although these signatures have not been fully translated to clinical practice as yet, they herald the promise of an improvement in breast cancer treatment decision-making. It should be noted, however, that most of the signatures developed to date seem to have discriminatory power almost restricted to oestrogen receptor-positive disease. This review addresses the contribution of gene expression profiling to our understanding of breast cancer and its clinical management and what has yet to be done for these classifiers to be incorporated in clinical practice. PMID- 19219437 TI - [Open letter from the German Society of Cytology: to the German Society of Gynecology and Obstetrics]. PMID- 19219439 TI - Loop ileostomy versus loop colostomy for fecal diversion after colorectal or coloanal anastomosis: a meta-analysis. AB - BACKGROUND: Sphincter-saving surgery for the treatment of middle and low rectal cancer has spread considerably when total mesorectal excision became standard treatment. In order to reduce leakage-related complications, surgeons often perform a derivative stoma, a loop ileostomy (LI), or a loop colostomy (LC), but to date, there is no evidence on which is the better technique to adopt. METHODS: We performed a systematic review and meta-analysis of all randomized controlled trials until 2007 and observational studies comparing temporary LI and LC for temporary decompression of colorectal and/or coloanal anastomoses. Clinically relevant events were grouped into four study outcomes: general outcome measures: dehydratation and wound infection GOM construction of the stoma outcome measures: parastomal hernia, stenosis, sepsis, prolapse, retraction, necrosis, and hemorrhage closure of the stoma outcome measures: anastomotic leak or fistula, wound infection COM, occlusion and hernia functioning of the stoma outcome measures: occlusion and skin irritation. RESULTS: Twelve comparative studies were included in this analysis, five randomized controlled trials and seven observational studies. Overall, the included studies reported on 1,529 patients, 894 (58.5%) undergoing defunctioning LI. LI reduced the risk of construction of the stoma outcome measure (odds ratio, OR = 0.47). Specifically, patients undergoing LI had a lower risk of prolapse (OR = 0.21) and sepsis (OR = 0.54). LI was associated with an excess risk of occlusion after stoma closure (OR = 2.13) and dehydratation (OR = 4.61). No other significant difference was found for outcomes. CONCLUSION: Our overview shows that LI is associated with a lower risk of construction of the stoma outcome measures. PMID- 19219440 TI - Fracture of the anterior extremity of calcaneus together with calcaneocuboid joint dislocation. AB - The calcaneocuboid joint is very stable, and dislocation or fracture is rare. We report a very rare case of dislocation fracture at the anterior articular surface of the calcaneocuboid joint. This is, to our knowledge, the first report of calcaneus fracture. PMID- 19219441 TI - Osteochondrosis dissecans of the medial malleolus: a case report. AB - Osteochondrosis dissecans of the ankle accounts for approximately 4% of all osteochondrosis dissecans. Most osteochondrosis dissecans in the ankle is found in the talar dome. We report the case of osteochondrosis dissecans of the medial malleolus in a 46-year-old patient which has not been described yet. PMID- 19219442 TI - Treatment of lateral epicondilitis using three different local injection modalities: a randomized prospective clinical trial. AB - BACKGROUND: To determine the effectiveness of three different local injection modalities in the treatment of lateral epicondilitis. METHODS: In a prospective randomized study on lateral epicondilitis, 75 patients were divided into three equal groups A, B and C (n = 25) and were treated using three different method of local injection. The patients in group A were treated with local injection of a steroid (1 mL triamcinolone) combined with local anaesthetic (1 mL lidocaine), those in group B were treated with injection of local anaesthetic (1 mL lidocaine) combined with peppering technique and those in group C with local injection of a steroid (1 mL triamcinolone) combined with local anaesthetic (1 mL lidocaine) and peppering technique. The outcome was defined by measuring the elbow pain during the activity using a 10-cm visual analogue scale (VAS) and satisfaction with the treatment using a scoring system based on the criteria of the Verhaar et al. at 3 weeks and 6 months after the injection and compared with the pre-treatment condition. RESULTS: There were significant (P = 0.006) differences in the successful outcomes between the three groups at 6 months. In group C in which local steroid + peppering injection technique were used; excellent results were obtained in 84% of patients comparing to 36% and 48% for patients in groups A and B, respectively. The successful outcomes were statistically higher in group C comparing to group A (P = 0.002) and group B (P = 0.011). In all groups, there was a significantly lower pain (VAS) at the 3-week and 6-month follow-ups comparing to the pre-treatment condition. VAS measured at 6-month follow-up were significantly lower in group C comparing to other groups (P = 0.002). CONCLUSION: In the treatment of lateral epicondilitis, combination of corticosteroid injections with peppering is more effective than corticosteroid injections or peppering injections alone and produces better clinical results. PMID- 19219443 TI - Simultaneous double dislocation of the interphalangeal joint in a finger. AB - Isolated traumatic dislocation of the distal and proximal interphalangeal joints of the fingers is a common occurrence. However, simultaneous dislocation of both interphalangeal joints in a single finger appears to be a rare event. We report five cases of simultaneous dislocation of both interphalangeal joints in a single finger with a review of the literature. PMID- 19219444 TI - Postmyomectomy sonographic imaging: uterus remodeling and scar repair. AB - PURPOSE: Description and evaluation of the sonographic changes at the site of the myomectomy and follow-up of the healing process. METHODS: Twelve patients with intramural fibroids underwent myomectomy. Pre- and postoperative ultrasound examinations included measurements of uterine volume and the largest diameters of the largest fibroid and the myomectomy scar. RESULTS: The mean uterine preoperative volume was 773 cm(3) (range 271-1,343) (SD 285). It decreased postoperatively to 318 cm(3) (range 98-756) (SD 192) and 185 cm(3) (range 78-420) (SD 96) in the immediate and late scans, respectively. The mean size of the largest diameter of the largest fibroid was 10 cm (range 7-15) (SD 2.3). The largest diameter of the immediate postoperative myomectomy site was 4.5 cm (range 2.2-8) (SD 1.8) decreasing later to 2.4 cm (range 0-6) (SD 1.6). CONCLUSION: The postoperative sonographic findings following myomectomy demonstrates a solid finding that may mistakenly be interpreted as the remaining fibroid that shrinks gradually. The finding is probably the result of changes in hyperplastic myometrial tissue, focal small hematomata and suture material. PMID- 19219445 TI - Severity of pain and circadian changes in uterine artery blood flow in primary dysmenorrhea. AB - OBJECTIVE: To investigate the severity of pain and circadian changes in uterine artery blood flow in primary dysmenorrhea cases. MATERIALS AND METHODS: The study included 27 cases diagnosed as primary dysmenorrhea and 25 individuals who had no dysmenorrhea complaint. Bilateral uterine artery systole/diastole rates (S/D), pulsatility indices (PI) and resistance indices (RI) of all cases were measured using transvaginal colour Doppler at 12.00-02.00 p.m. and 12.00-02.00 a.m. Severity of pain was evaluated in dysmenorrhea cases at the same hours using a verbal pain assessment scale. FINDINGS: Doppler measurements of dysmenorrhea cases conducted at 12.00-02.00 p.m. showed right uterine artery S/D: 3.37 +/- 0.26, RI: 0.73 +/- 0.07, PI: 2.38 +/- 0.34 and left uterine artery S/D: 3.33 +/- 0.37, RI: 0.74 +/- 0.14, PI: 2.41 +/- 0.15, while measurements carried out at 12.00-02.00 a.m. showed right uterine artery S/D: 3.88 +/- 0.12, RI: 0.87 +/- 0.14, PI: 2.94 +/- 0.21 and left uterine artery S/D: 3.90 +/- 0.27, RI: 0.92 +/- 0.12, PI: 2.93 +/- 0.21. Comparisons revealed significantly higher Doppler indices at night (P < 0.05) than in the day in dysmenorrhea cases. There was not any significant circadian difference in individuals who did not have dysmenorrhea (P > 0.05). Pain score in the verbal pain assessment of dysmenorrhea cases was found 3.6 +/- 1.4 in the day and 5.8 +/- 1.7 at night. The difference was found significant (P < 0.05). CONCLUSION: Uterine artery blood flow is reduced at night in dysmenorrhea cases. In correlation with this, the cases feel more pain at night. Our results may be important on the planning of working hours and their quality of life. PMID- 19219446 TI - Patterns and success of fetal programming among women with low and extremely low pre-pregnancy BMI. AB - PURPOSE: To estimate the frequency of fetal programming phenotypes among women with low BMI and the success of these programming patterns-to determine if small for gestational age (SGA) is a biologically adaptive mechanism to improve chances for infant survival. METHODS: We examined the frequency of fetal programming phenotypes: SGA, large for gestational age (LGA), and adequate for gestational age (AGA) among 1,063,888 singleton live births from 1978 to 1997. We also estimated the success of fetal programming phenotypes using neonatal death as the primary study outcome. RESULTS: Underweight gravidas with AGA and LGA babies had elevated risk of neonatal mortality when compared to normal weight mothers, while the risk for neonatal mortality among mothers with SGA babies was reduced. CONCLUSIONS: The variation in relative degrees of fetal programming patterns and success observed suggests that underweight mothers are more likely to succeed in programming SGA fetuses rather than any other phenotype. PMID- 19219447 TI - The fetal cheek-to-cheek diameter and abdominal circumference: are they correlated? AB - OBJECTIVE: The cheek-to-cheek diameter (CCD) has been shown to be an indicator of subcutaneous tissue mass in the fetus. However, the correlation between CCD and the abdominal circumference (AC) has not been investigated yet. The objective of the present study was to demonstrate whether a correlation exists between fetal CCD, AC, estimated fetal weight (EFW), and the 1 h, 50 g, glucose challenge test (GCT) levels in patients with and without gestational diabetes mellitus. METHODS: A prospective, institutional review board approved study was performed. The CCD was obtained as part of the ultrasound for obstetric interval growth scans and biophysical profiles. Exams were performed during the third trimester. The CCD was obtained on a coronal view of the fetal face, at the level of the nostrils and lips. Patients were enrolled between November 2005 and May 2006. Pearson correlation coefficient and linear regression modeling were used as appropriate. RESULTS: Eighty-three patients were enrolled, 29 (33%) of them were diabetic. The mean gestational age is 34.8 +/- 3 weeks and the mean maternal age is 29.9 +/- 5.1. A significant linear association was found between CCD and EFW (Pearson coefficient of correlation being 0.51, P = 0.01). The Pearson correlation coefficient of the relationship between the CCD and AC was 0.47 (P = 0.01). Using a linear regression model, controlling for gestational age at performance of the ultrasound, the association between CCD and EFW remained significant (P = 0.021). There were no significant differences between diabetic and non-diabetic patients regarding the CCD (6.2 +/- 0.9 vs. 6.3 +/- 0.9 respectively, P = 0.669) or the EFW (2,527.9 +/- 705 vs. 2,645 +/- 760 g). While AC was significantly correlated with the GCT levels (Pearson coefficient of correlation = 0.46, P = 0.024), no such correlation was demonstrated for CCD (Pearson correlation coefficient = 0.23, P = 0.160). CONCLUSIONS: The cheek-to-cheek diameter is significantly correlated to the abdominal circumference and the estimated fetal weight. However, the abdominal circumference has a tighter correlation with the glucose challenge test. PMID- 19219448 TI - Spectroscopic, kinetic and dosimetric features of the radical species produced after radiodegradation of solid triclosan. AB - In the present work, spectroscopic, kinetic and dosimetric features of the radicalic intermediates produced after gamma irradiation at room temperature of solid triclosan (2,4,4-trichloro-2-hydroxydiphenyl ether; TCS) were investigated by means of electron spin resonance spectroscopy (ESR) at various temperatures. The same material was also irradiated with UV light, and an ESR spectrum very similar to that obtained for gamma-irradiated TCS, was recorded. The ESR spectrum of TCS is characterized by an unresolved doublet with resonance lines split into other doublets. An evaluation technique based on variations of four assigned peak to-peak amplitudes and signal intensity was adopted, to monitor the evolution of the spectrum under different experimental conditions. Radicals of one type were proposed to be created upon irradiation exhibiting decays via intra-track and inter-track recombination reactions with activation energies of 43 +/- 2 and 139 +/- 6 kJ/mol, respectively. A radical exhibiting axial g anisotropy and interacting with two un-equivalent protons was found to describe the experimental spectrum well. The sensitivity of TCS to gamma radiation was high (G = 0.12) suggesting TCS to be a suitable dosimetric material in measuring normal and accidental radiation doses in the range of (1-25 kGy). PMID- 19219449 TI - Peroxisomes: 40 years of histochemical staining, personal reminiscences. AB - The historical circumstances that led to the discovery of the 3,3'-diamino benzidine (DAB) method for staining of peroxisomes 40 years ago are reviewed. In the course of studies on the uptake and absorption of horse radish peroxidase in mammalian liver, in sections incubated for detection of peroxidase activity in DAB, it was noted that peroxisomes also stained positively for peroxidase activity. Subsequently, it was revealed that the peroxidatic activity of catalase, which is abundantly present in peroxisomes, is responsible for that staining. This notion was confirmed in quantitative biochemical studies with crystalline beef liver catalase and in tracer studies using catalase as an ultrastructural tracer. The application of the DAB method led to the discovery of peroxisomes as a ubiquitous eukaryotic cell organelle, attracting great interest in their investigation in biomedical research. PMID- 19219450 TI - Biogenesis of peroxisomes and mitochondria: linked by division. AB - Peroxisomes and mitochondria are metabolically linked organelles, which are crucial to human health and development. The search for components involved in their dynamics and maintenance led to the interesting finding that mitochondria and peroxisomes share components of their division machinery. Recently, it became clear that this is a common strategy used by mammals, fungi and plants. Furthermore, a closer interrelationship between peroxisomes and mitochondria has been proposed, which might have an impact on functionality and disease conditions. Here, we briefly highlight the major findings, views and open questions concerning peroxisomal formation, division, and interrelationship with mitochondria. PMID- 19219451 TI - Pre-dive normobaric oxygen reduces bubble formation in scuba divers. AB - Oxygen pre-breathing is routinely employed as a protective measure to reduce the incidence of altitude decompression sickness in aviators and astronauts, but the effectiveness of normobaric oxygen before hyperbaric exposure has not been well explored. The objective of this study was to evaluate the effect of 30-min normobaric oxygen (O(2)) breathing before diving upon bubble formation in recreational divers. Twenty-one subjects (13 men and 8 women, mean age (SD) 33 +/ 8 years) performed random repetitive open-sea dives (surface interval of 100 min) to 30 msw for 30 min with a 6-min stop at 3 msw under four experimental protocols: "air-air" (control), "O(2)-O(2)", "O(2)-air" and "air-O(2)" where "O(2)" corresponds to a dive with oxygen pre-breathing and "air" a dive without oxygen administration. Post-dive venous gas emboli were examined by means of a precordial Doppler ultrasound. The results showed decreased bubble scores in all dives where preoxygenation had taken place (p < 0.01). Oxygen pre-breathing before each dive ("O(2)-O(2)" condition) resulted in the highest reduction in bubble scores measured after the second dive compared to the control condition ( 66%, p < 0.05). The "O(2)-air" and "air-O(2) "conditions produced fewer circulating bubbles after the second dive than "air-air" condition (-47.3% and 52.2%, respectively, p < 0.05) but less bubbles were detected in "air-O(2) "condition compared to "O(2)-air" (p < 0.05). Our findings provide evidence that normobaric oxygen pre-breathing decreases venous gas emboli formation with a prolonged protective effect over time. This procedure could therefore be beneficial for multi-day repetitive diving. PMID- 19219452 TI - Caveolin, GLUT4 and insulin receptor protein content in human arm and leg muscles. AB - Recent evidence suggests that insulin sensitivity is relatively better preserved in arm muscle than in leg muscle in both healthy controls and type 2 DM, based on measurements of basal and insulin-mediated glucose clearance performed simultaneously in the two sets of muscles. It has also been reported that glucose uptake rates are higher in arm compared to leg muscles in the fasted state during normo-insulinaemia. However, the mechanism(s) for this are unknown. Currently, no information is available on the content of glucose transport proteins between arm and leg muscles. Therefore, we compared four proteins, Caveolin-1 (Cav-1), Caveolin-3 (Cav-3), GLUT4, and IR-beta, each of which plays an important role in regulating glucose transport between arm and leg muscles using muscle samples that were obtained from the deltoid (DEL) and vastus lateralis (VAS) of 14 male college pentathlon athletes before and after two swimming trials performed over 100 and 1,500 m. In the present study, we have shown the levels of Cav-1, -3, GLUT4, and IR-beta measured together for the first time in human arm and leg muscles. There was no difference in the levels of these proteins between arm and leg muscles. Cav-3, GLUT4, and IR-beta were unchanged from the resting levels after both exercise trials in DEL, while Cav-1 was increased (17%) at the end of the longer swim trial. In contrast, all measurements of Cav-1, -3, GLUT4, and IR beta after the 1,500 m swim trial in VAS were increased, by 120, 46, 123, and 60%, respectively. These data imply that there was no functional difference in glucose transport capacity between arm and leg muscles in highly trained pentathlon athletes in the resting state. Although Cav-3, GLUT4, and IR-beta were unchanged from the resting levels at the end of both exercise trials in DEL, all measures, including Cav-1, increased after the 1,500 m swim trial in VAS. PMID- 19219453 TI - Acquired (idiopathic) intradermal nevus with junctional activity presenting as clitoromegaly in a child: report of a case. AB - Clitoromegaly (macroclitoris) can either be congenital or acquired. Intradermal nevus presenting as clitoromegaly in a child is extremely rare. Extensive Medline search has failed to reveal any published article on this. To the best of our knowledge, we report the first patient with this presentation in a 6-year-old child. PMID- 19219454 TI - Cutaneous signs are important in the diagnosis of the rare neoplasia syndrome Carney complex. AB - We describe a 15-year-old boy who presented with a stroke. Brain MRI imaging showed thalamic and multiple cerebral infarcts. An echocardiogram revealed multiple atrial masses, which were resected. Histological examination confirmed multiple atrial myxomas. Further clinical examination of the patient revealed subtle buccal and peri-oral lentigenes. The diagnosis of Carney complex was made clinically. The patient was subsequently diagnosed with testicular seminomas and a cutaneous angiomyxoma. Genetic investigation revealed a pathological mutation in the PRKAR1A gene. We review the reported manifestations and presentations of Carney complex, along with current diagnostic guidelines. We emphasise the importance of recognising the cutaneous manifestations of this rare autosomal dominantly inherited neoplasia syndrome. PMID- 19219455 TI - Useful methods in preventing accidental falls from the bed in children at the emergency department. AB - The aim of this study was to analyze the general characteristics of children in the pediatric emergency department (PED) who accidentally fall off the crib and to establish useful preventive measures. This prospective research analyzed pediatric patients who accidentally fell off their beds in the observational unit (OU) of the PED from July 2005 to June 2006 (first period). From July 2006 to February 2007 (second period), the causes of children falling off the crib in the first year were analyzed and five related preventive methods were instituted in the OU. From July 2007 to March 2008 (third period), the preventive methods were enhanced to achieve zero-event of accidental falls in the PED. The differences between patients falling off the bed among the three periods were then compared. This study collected 7,281 children admitted to the OU during the first period and recorded 15 cases of accidental falls. After performing the preventive methods in 6,232 patients in the second period, three events of accidental falls were noted. In the third period, there was no accident in the 5,225 patients admitted to the PED. Comparing the occurrences of children falling off the bed among the three periods, accidental falls significantly decreased in the third period (p < 0.001). Effective methods can be instituted to prevent children from falling off the bed, especially in the PED. PMID- 19219456 TI - Modelling the drought impact on monoterpene fluxes from an evergreen Mediterranean forest canopy. AB - In many ecosystems drought cycles are common during the growing season but their impact on volatile monoterpene emissions is unclear. Therefore, we aimed to develop and evaluate a process-based modelling approach to explore the explanatory power of likely mechanisms. The biochemically based isoprene and monoterpene emission model SIM-BIM2 has been modified and linked to a canopy model and a soil water balance model. Simulations are carried out for Quercus ilex forest sites and results are compared to measured soil water, photosynthesis, terpene-synthase activity, and monoterpene emission rates. Finally, the coupled model system is used to estimate the annual drought impact on photosynthesis and emission. The combined and adjusted vegetation model was able to simulate photosynthesis and monoterpene emission under dry and irrigated conditions with an R(2) of 0.74 and 0.52, respectively. We estimated an annual reduction of monoterpene emission of 67% for the extended and severe drought period in 2006 in the investigated Mediterranean ecosystem. It is concluded that process-based ecosystem models can provide a useful tool to investigate the involved mechanisms and to quantify the importance of specific environmental constraints. PMID- 19219457 TI - Expression of parasite virulence at different host population densities under natural conditions. AB - It has recently been suggested that the expression of parasite virulence depends on host population density, such that infected hosts have a higher sensitivity to density, and thus reach their carrying capacity earlier than uninfected hosts. In this scenario, parasite-induced reduction in fitness (i.e., virulence) increases with host density. We tested this hypothesis experimentally, using outdoor mesocosm populations of Daphnia magna infected by the microsporidian Octosporea bayeri. Contrary to the prediction, virulence was independent of host density. In a competition experiment with initial prevalence of 50%, O. bayeri reduced the competitive ability of infected Daphnia within the asexual growth phase independent of initial host population density. In an additional experiment we set up populations with 100% and 0% prevalence and followed their population dynamics over the whole season. Consistent with the competition experiment, we found no difference in population dynamics within the asexual growth phase of the host, suggesting that infected hosts are not more sensitive to density than uninfected hosts. The additional experiment, however, included more than the initial growth phase as did the competition experiment. Eventually, after 100 days, 100% infected populations assumed a reduced carrying capacity compared to uninfected populations. We identify and discuss three reasons for the discrepancy between our experiment and the predictions. PMID- 19219458 TI - Specific bottom-up effects of arbuscular mycorrhizal fungi across a plant herbivore-parasitoid system. AB - The majority of plants are involved in symbioses with arbuscular mycorrhizal fungi (AMF), and these associations are known to have a strong influence on the performance of both plants and insect herbivores. Little is known about the impact of AMF on complex trophic chains, although such effects are conceivable. In a greenhouse study we examined the effects of two AMF species, Glomus intraradices and G. mosseae on trophic interactions between the grass Phleum pratense, the aphid Rhopalosiphum padi, and the parasitic wasp Aphidius rhopalosiphi. Inoculation with AMF in our study system generally enhanced plant biomass (+5.2%) and decreased aphid population growth (-47%), but there were no fungal species-specific effects. When plants were infested with G. intraradices, the rate of parasitism in aphids increased by 140% relative to the G. mosseae and control treatment. When plants were associated with AMF, the developmental time of the parasitoids decreased by 4.3% and weight at eclosion increased by 23.8%. There were no clear effects of AMF on the concentration of nitrogen and phosphorus in plant foliage. Our study demonstrates that the effects of AMF go beyond a simple amelioration of the plants' nutritional status and involve rather more complex species-specific cascading effects of AMF in the food chain that have a strong impact not only on the performance of plants but also on higher trophic levels, such as herbivores and parasitoids. PMID- 19219459 TI - Summer temperature dependency of larch budmoth outbreaks revealed by Alpine tree ring isotope chronologies. AB - Larch budmoth (LBM, Zeiraphera diniana Gn.) outbreaks cause discernable physical alteration of cell growth in tree rings of host subalpine larch (Larix decidua Mill.) in the European Alps. However, it is not clear if these outbreaks also impact isotopic signatures in tree-ring cellulose, thereby masking climatic signals. We compared LBM outbreak events in stable carbon and oxygen isotope chronologies of larch and their corresponding tree-ring widths from two high elevation sites (1800-2200 m a.s.l.) in the Swiss Alps for the period AD 1900 2004 against isotope data obtained from non-host spruce (Picea abies). At each site, two age classes of tree individuals (150-250 and 450-550 years old) were sampled. Inclusion of the latter age class enabled one chronology to be extended back to AD 1650, and a comparison with long-term monthly resolved temperature data. Within the constraints of this local study, we found that: (1) isotopic ratios in tree rings of larch provide a strong and consistent climatic signal of temperature; (2) at all sites the isotope signatures were not disturbed by LBM outbreaks, as shown, for example, by exceptionally high significant correlations between non-host spruce and host larch chronologies; (3) below-average July to August temperatures and LBM defoliation events have been coupled for more than three centuries. Dampening of Alps-wide LBM cyclicity since the 1980s and the coincidence of recently absent cool summers in the European Alps reinforce the assumption of a strong coherence between summer temperatures and LBM defoliation events. Our results demonstrate that stable isotopes in tree-ring cellulose of larch are an excellent climate proxy enabling the analysis of climate-driven changes of LBM cycles in the long term. PMID- 19219460 TI - Multifaceted determinants of host specificity in an aphid parasitoid. AB - The host specificity of insect parasitoids and herbivores is thought to be shaped by a suite of traits that mediate host acceptance and host suitability. We conducted laboratory experiments to identify mechanisms shaping the host specificity of the aphid parasitoid Binodoxys communis. Twenty species of aphids were exposed to B. communis females in microcosms, and detailed observations and rearing studies of 15 of these species were done to determine whether patterns of host use resulted from variation in factors such as host acceptance or variation in host suitability. Six species of aphids exposed to B. communis showed no signs of parasitism. Four of these species were not recognized as hosts and two effectively defended themselves from attack by B. communis. Other aphid species into which parasitoids laid eggs had low suitability as hosts. Parasitoid mortality occurred in the egg or early larval stages for some of these hosts but for others it occurred in late larval stages. Two hypotheses explaining low suitability were investigated in separate experiments: the presence of endosymbiotic bacteria conferring resistance to parasitoids, and aphids feeding on toxic plants. An association between resistance and endosymbiont infection was found in one species (Aphis craccivora), and evidence for the toxic plant hypothesis was found for the milkweed aphids Aphis asclepiadis and Aphis nerii. This research highlights the multifaceted nature of factors determining host specificity in parasitoids. PMID- 19219461 TI - Biochemical tracers reveal intra-specific differences in the food webs utilized by individual seabirds. AB - Food web structure regulates the pathways and flow rates of energy, nutrients, and contaminants to top predators. Ecologically and physiologically meaningful biochemical tracers provide a means to characterize and quantify these transfers within food webs. In this study, changes in the ratios of stable N isotopes (e.g., delta(15)N), fatty acids (FA), and persistent contaminants were used to trace food web pathways utilized by herring gulls (Larus argentatus) breeding along the shores of the St Lawrence River, Canada. Egg delta(15)N values varied significantly among years and were used as an indicator of gull trophic position. Temporal trends in egg delta(15)N values were related to egg FA profiles. In years when egg delta(15)N values were greater, egg FA patterns reflected the consumption of more aquatic prey. Egg delta(15)N values were also correlated with annual estimates of prey fish abundance. These results indicated that temporal changes in aquatic prey availability were reflected in the gull diet (as inferred from ecological tracer profiles in gull eggs). Analysis of individual eggs within years confirmed that birds consuming more aquatic prey occupied higher trophic positions. Furthermore, increases in trophic position were associated with increased concentrations of most persistent organic contaminants in eggs. However, levels of highly brominated polybrominated diphenyl ether congeners, e.g, 2,2',3,3',4,4',5,5',6,6'-decabromoDE (BDE-209), showed a negative relationship with trophic position. These contrasting findings reflected differences among contaminant groups/homologs in terms of their predominant routes of transfer, i.e., aquatic versus terrestrial food webs. High trophic level omnivores, e.g., herring gulls, are common in food webs. By characterizing ecological tracer profiles in such species we can better understand spatial, temporal, and individual differences in pathways of contaminant, energy, and nutrient flow. PMID- 19219462 TI - Novel weapons and invasion: biogeographic differences in the competitive effects of Centaurea maculosa and its root exudate (+/-)-catechin. AB - Recent studies suggest that the invasive success of Centaurea maculosa may be related to its stronger allelopathic effects on native North American species than on related European species, one component of the "novel weapons" hypothesis. Other research indicates that C. maculosa plants from the invasive range in North America have evolved to be larger and better competitors than conspecifics from the native range in Europe, a component of the "evolution of increased competitive ability" hypothesis. These hypotheses are not mutually exclusive, but this evidence sets the stage for comparing the relative importance of evolved competitive ability to inherent competitive traits. In a competition experiment with a large number of C. maculosa populations, we found no difference in the competitive effects of C. maculosa plants from North America and Europe on other species. However, both North American and European C. maculosa were much better competitors against plants native to North America than congeners native to Romania, collected in areas where C. maculosa is also native. These results are consistent with the novel weapons hypothesis. But, in a second experiment using just one population from North America and Europe, and where North American and European species were collected from a broader range of sites, competitive interactions were weaker overall, and the competitive effects of C. maculosa were slightly stronger against European species than against North American species. Also consistent with the novel weapons hypothesis, (+/-)-catechin had stronger effects on native North American species than on native European species in two experiments. Our results suggest that the regional composition of the plant communities being invaded by C. maculosa may be more important for invasive success than the evolution of increased size and competitive ability. PMID- 19219464 TI - Urbanisation induces early flowering: evidence from Platanus acerifolia and Prunus cerasus. AB - The effect of towns on plant phenology, i.e. advancement of spring development compared with a rural environment, via the urban heat island (UHI) phenomenon, has been shown for many towns in many countries. This work combines experimental and observational methodology to provide a better and deeper view of climatic habitat in an urban context with a view to understanding the relationship between plant development and urban climate on the intra-urban scale (by taking into account town structure). A dense network of 17 meteorological stations was set up in Rennes, France, enabling us to identify and quantify climatic changes associated with the UHI. Meanwhile, phenological observations were made during early spring (March and April) in 2005 on Platanus acerifolia and Prunus cerasus to study the relationship between climatic and phenological data. The results show that there is both a climatic gradient and a developmental gradient corresponding to the type of urbanisation in the town of Rennes. The town influences plant phenology by reducing the diurnal temperature range and by increasing the minimum temperature as one approaches the town centre. The influence of ground cover type (plants or buildings) on development is also shown. The developmental phases of preflowering and flowering are influenced to differing extents by climatic variables. The period during which climatic variables are effective before a given developmental phase varies considerably. The preflowering phases are best correlated with the mean of the minimum air temperature for the 15-day period before the observation, whereas flowering appears to be more dependent on the mean of the daily diurnal temperature range for the 8 days preceding the observation. PMID- 19219463 TI - Minimal change disease with IgM+ immunofluorescence: a subtype of nephrotic syndrome. AB - Immunoglobulin (Ig) M nephropathy is defined by electron-dense mesangial deposits and mesangial IgM visible by immunofluorescence (IF) without other histopathologic and immunofluorescent microscopic abnormalities. Certain patients have only immuno-positive (IgM+) IF. Children presenting with steroid-dependent or steroid-resistant nephrotic syndrome have a high prevalence of IgM+ IF with or without electron-dense deposits. We reviewed the clinical course of children with steroid-dependent or steroid-resistant nephrotic syndrome who underwent renal biopsy at Texas Children's Hospital from 1989 to 2006 to further characterize IgM+ IF in children with nephrotic syndrome. Of the 55 children with steroid resistant or -dependent minimal change disease (MCD), 23 had IgM+ IF. Of these 23 children, 61% had microscopic hematuria at presentation, 48% (11/23) were steroid dependent, and 48% (11/23) steroid-resistant (one underwent biopsy prior to steroid therapy). We compared the efficacy of adjuvant treatment with cyclophosphamide and cyclosporine: 18% initially treated with cyclophosphamide obtained remission, while 55% had no response; 83% obtained subsequent remission with cyclosporine. Of those initially treated with cyclosporine, 88% obtained complete or partial remission. IgM+ IF may be surrogate marker for the severity of MCD. Based on our results, children with MCD and IgM+ IF have a better response to cyclosporine than cyclophosphamide. PMID- 19219465 TI - Evaluation of Golestan Province's Early Warning System for flash floods, Iran, 2006-7. AB - Golestan province located in NE Iran is well known for deadly flash floods. This study aimed to evaluate the region's Early Warning System (EWS) for flash floods. We used an adapted version of the questionnaire developed by the United Nations International Strategy for Disaster Reduction. We reviewed documents on the EWS of Golestan, and conducted a qualitative study comprising interviews with experts and affected people in Kalaleh and Minoodasht. Results were discussed by an expert panel. Regarding risk knowledge, there was a hazard map at Provincial Disaster Taskforce (PDT) drawn by the provincial Office for Water Resource Management, but no risk analysis was available. Local people were aware of their exposure to flooding, but not aware of the existence of a hazard map and their vulnerability situation. In terms of monitoring and warning, PDT faced serious limitations in issuing Early Warnings, including (1) an inability to make point predictions of rainfall, and (2) the absence of a warning threshold. Dissemination and communication issued by the Meteorological Office followed a top-to-bottom direction. The contents were neither clearly understood by other institutions nor reached the potential recipients within an appropriate time frame. There was a need for a comprehensive response plan with adequate exercises, and no evaluation framework existed. Golestan EWS is in dire need of improvement. To fill in the gaps ensuring local people receive timely warning, we propose a community-based model called "Village Disaster Taskforce" (VDT) in which individual villages act as operational units, but interlinked with other villages and PDT. PMID- 19219466 TI - An improved estimation of daily clear-sky biologically EER from broadband global solar radiation. AB - To establish a relation between biologically effective erythemal radiation (EER) and global solar radiation, the hourly and daily clear-sky broadband (310-2,800 nm) global solar radiation (G) and spectral ultraviolet radiation incident on a horizontal surface at Esfahan, Iran (32 degrees 37'N, 51 degrees 40'E) were measured during the period 2001-2005. Good correlations at statistically significant levels between the daily values of EER and the daily G were found. The seasonal variability of EER/G is also discussed and the correction factors are determined for inclusion of vertical column ozone and solar zenith angle (SZA) cycles. The comparison of the estimated daily EER against the independent observed EER revealed that under clear sky conditions the estimations are accurate to 10% or better over SZA of 10-60 degrees and column ozone of 250-350 Dobson. The comparison of the results with the similar works that have used shorter period of experimental data showed more accurate estimates. The deduced relations could be used to a rough estimate of the daily EER from G in arid climate regions, where there is no measured UV radiation or there are instrumental and other difficulties encountered in measuring UV radiation. PMID- 19219468 TI - Surgical resection without dural reconstruction of a lumbar meningioma in an elderly woman. AB - Meningiomas of the spine occur in the thoracic spine in approximately 80%, followed in frequency by the cervical and lumbar regions. The treatment of spinal meningiomas is complete surgical resection. As intraspinal meningiomas are almost always adherent to the dura, extensive dural resection or diathermic treatment of the dural attachment is usually performed to prevent tumor recurrence. The authors present the case of lumbar spinal meningioma in 82-year-old woman. Successful resection with preservation of the dura mater using the technique of Saito et al. (Spine 26:1805-1808, 2001) is described: After lumbar laminectomy a small incision was made in the surface of the spinal dura. The dura mater was separated into its inner and outer layers, and the tumor was resected with inner layer alone, preserving the outer layer. The outer layer is simply closed to achieve a watertight seal. The pathologic diagnosis was metaplastic (osseous) meningioma. Almost full recovery of the neurologic deficit was attained. Neither complication nor tumor recurrence has occurred in the 5 years since surgery. Dural preservation during surgical resection of spinal meningioma obviates the need for dural reconstruction and should reduce surgical morbidity. However, the patient should be followed long-term to watch for recurrence. PMID- 19219467 TI - Surgical assessment of the proximal thoracic curve in adolescent idiopathic scoliosis. AB - Existing predictive signs as available in current literature may miss potential proximal thoracic (PT) curve deterioration and shoulder imbalance, following selective main thoracic (MT) curve correction in adolescent idiopathic scoliosis (AIS). The present study is an attempt to evaluate and complement these signs, through a retrospective study of 56 AIS patients who underwent correction and fusion from 1986 till 2003 with follow-up 4-16 years. Forty-nine had fusion of MT curve, 7 of MT and PT. Cotrel-Dubousset instrumentation in 45, Luque in 12. Preoperative data: MT 50 degrees (40 degrees -80 degrees), PT 25 degrees (0 degrees -50 degrees), shoulder elevation from -4 cm (right) to 2 cm (left), clavicle angle from -14 degrees to 5 degrees , PT bending correction from 0 to 100% and T1 tilt from -15 degrees to 14 degrees . We introduced the first rib index (FRI), i.e., the difference between the diameter of right and left first rib arch as a percentage of the sum of both diameters, averaging from -22.7 to 14.3%. (Minus signs refer to or predict right, while positive left shoulder elevation.) Evaluation included all predictive parameters as related principally to postoperative left shoulder elevation > or =1 cm, patient satisfaction and surgeon fulfillment. Postoperative correction MT curve 53% (23-83%) and PT 35% (0 100%). One progressive paraplegic started 40 min following normal wake-up test. Immediate decompression, full recovery. Three cases with wound infection recovered after late removal of instrumentation. Loss of correction > or =10 degrees in five. Fifteen had postoperative persisting left shoulder elevation > or =1 cm. Seven of these expressed dissatisfaction. Statistically FRI proved valuable predictive factor always in combination with previously described signs. We concluded that a postoperative left shoulder elevation >/=2 cm is a potential cause of dissatisfaction and may be prevented with thorough validation of all predictive signs, principally the FRI. PMID- 19219469 TI - Maxillomandibular osteomyelitis in osteopetrosis. Report of a case and review of the literature. AB - INTRODUCTION: Osteopetrosis is a rare hereditary group of bony displasias. They range from a devastating metabolic disease (including severe malignant infantile osteopetrosis) to other conditions with a more benign phenotype (autosomal dominant osteopetrosis I and II). CASE REPORT: Several case reports have been published of infectious complications affecting the mandible but maxillary affectation is quite rare. We present the case of a 23-year-old woman with a malignant recessive form of osteopetrosis complicated by repeated episodes of osteomyelitis (caused by actinomyces) of the mandible and maxilla. PMID- 19219470 TI - Association between migraine and a functional polymorphism at the dopamine beta hydroxylase locus. AB - Migraine is a common neurological disorder with a significant genetic component. Although a number of linkage and association studies have been undertaken, the number and identity of all migraine susceptibility genes has yet to be defined. The existence of dopaminergic hypersensitivity in migraine has been recognised on a pharmacological basis and some studies have reported genetic association between migraine and dopamine-related gene variants. Our laboratory has previously reported association of migraine with a promoter STR marker in the dopamine beta hydroxylase (DBH) gene. In the present study, we analysed two additional DBH markers in two independent migraine case-control cohorts. These two markers are putative functional SNPs, one within the promoter (-1021C-->T) and another SNP (+1603C-->T) in exon 11 of the DBH gene. The results showed a significant association for allelic and genotypic frequency distribution between the DBH marker in the promoter and migraine in the first (P = 0.004 and P = 0.012, respectively) and the second (P = 0.013 and P = 0.031, respectively) tested cohorts. There was no association observed between either genotype and/or allelic frequencies for the DBH marker located in exon 11 and migraine (P > or = 0.05). The promoter DBH marker, reported associated with migraine in this study, has been shown to affect up to 52% of plasma DBH activity. Varying DBH activity levels have been postulated to be involved in migraine process with an increase of dopamine, resulting from a lower DBH activity shown positively correlated with migraine severity. It is plausible that the functional promoter variant of DBH may play a role in the migraine disorder. PMID- 19219471 TI - Yersinia enterocolitica and Y. enterocolitica-like species in clinical stool specimens of humans: identification and prevalence of bio/serotypes in Finland. AB - This study investigated the prevalence of Yersinia enterocolitica (YE) bio/serotypes and YE-like species in clinical stool specimens. The special aim was to find the best methods for accurate identification of YE species and, further, pathogenic strains among YE isolates. Of the 41,848 specimens cultured in ten laboratories during a 12-month period, 473 Yersinia strains were isolated from 462 patients. The strains were identified by 21 biochemical tests, serotyping, colony morphology, as well as by 16S rRNA and gyrB gene sequencing. The most prevalent Yersinia findings were YE biotype 1A (64% of the strains) and pathogenic bio/serotype 4/O:3 (16%). The cold-enrichment increased the number of all isolates, and 25% of the bio/serotype 4/O:3 and 2/O:9 strains were only found by cold-enrichment. In routine diagnostic laboratories, 50% of the YE-like species were identified as YE and in 26% the identification differed from that of the reference laboratory. The microscopic colony identification on CIN agar with positive CR-MOX test, combined with several biochemical tests, identified reliably the pathogenic YE bioserotypes and most YE BT 1A strains, but some strains of the YE-like species were so heterogenic that gene sequencing was the only way to identify them. PMID- 19219472 TI - Nontuberculous mycobacterial infections among patients suspected of pulmonary tuberculosis. AB - The purpose of this study was to present a retrospective analysis of the frequency of nontuberculous mycobacteria (NTM)-related pulmonary infections among the AFB-positive and/or culture-positive patients in the Warsaw region who were suspected of tuberculosis (TB) and hospitalized in the university hospital between 1999 and 2005. All the AFB-positive pulmonary samples were examined with a molecular method using the Amplicor MTB test (Roche) for detection of Mycobacterium tuberculosis complex, and all mycobacterial isolates were speciated by high performance liquid chromatography (HPLC) analysis of mycolic acids. Patients who met clinical, radiological, and bacteriological criteria of mycobacteriosis were classified according to the American Thoracic Society (ATS) guidelines for diagnosis of NTM related disease. Among the 445 smear-positive or/and culture-positive patients, 142 subjects (31.9%) were found to be infected with M. tuberculosis. Among 303 non-TB patients, mycobacteriosis was found in 27 (8.9%) subjects. The frequency of NTM-related lung disease as compared to the bacteriologically-confirmed lung TB was estimated at 1:5. The rapid, precise methods of NTM speciation are necessary for progress in diagnostics of NTM related diseases. PMID- 19219473 TI - A microbiological study of the self-cleaning potential of oily Arabian Gulf coasts. AB - BACKGROUND, AIM, AND SCOPE: Due to the active production and transport of crude oil in the Arabian Gulf region, the Arabian Gulf coasts are routinely polluted with oil. Therefore, such coasts have been subject of studies aiming at assessing the roles of indigenous microbial consortia in cleaning these environments. In the present study, epilithic microbial communities along Kuwait coasts were studied for their oil degradation potential. MATERIALS AND METHODS: Gravel particles coated with deep green biofilms were collected from four coastal sites in autumn, winter, and spring. Phototrophs in these consortia were determined in terms of their chlorophyll a contents and identified by their morphological characteristics. Total bacteria were counted microscopically and cultivable bacteria by the dilution plating method on nutrient agar as well as on inorganic medium containing oil as a sole source of carbon and energy. The bacterial community structures were also characterized and compared by denaturing gradient gel electrophoresis (DGGE). RESULTS: Epilithic biomass samples from the four sites in the three seasons were rich in diatoms and picocyanobacteria as well as total bacteria. Direct counting gave bacterial numbers per square centimeter gravel surface of 2 to 6 x 10(7) cells depending on the sampling site and season. Cultivable bacterial numbers on nutrient agar and crude oil as a sole source of carbon were 3 x 10(3) to 8 x 10(4) and 1 x 10(3) to 7 x 10(3) cells/cm(2) gravel surface, respectively. The DGGE profiles of epilithon biomass samples revealed major 16S rDNA bands that matched bands of pure oil-utilizing bacterial isolates. DISCUSSION: The microbial communities showed a degree of consistency in all sites and seasons. CONCLUSIONS: The microbial consortia coating gravel particles are potentially suitable tools for self-cleaning of oily Gulf coasts. They are rich in oil-utilizing bacteria whose activities are probably enhanced by oxygen produced by the phototrophic partners in the consortia. RECOMMENDATIONS AND PERSPECTIVES: The combination of conventional microbiological analysis with molecular approaches gives an enhanced idea about natural microbial communities especially those with environmental application potential. PMID- 19219474 TI - Trihalomethane formation during water disinfection in four water supplies in the Somes river basin in Romania. AB - BACKGROUND, AIM AND SCOPE: After the discovery of chloroform in drinking water, an extensive amount of work has been dedicated to the factors influencing the formation of halogenated disinfections by-products (DBPs). The disinfection practice can vary significantly from one country to another. Whereas no disinfectant is added to many water supplies in Switzerland or no disinfectant residual is maintained in the distribution system, high disinfectant doses are applied together with high residual concentrations in the distribution system in other countries such as the USA or some southern European countries and Romania. In the present study, several treatment plants in the Somes river basin in Romania were investigated with regard to chlorine practice and DBP formation (trihalomethanes (THMs)). Laboratory kinetic studies were also performed to investigate whether there is a relationship between raw water dissolved organic matter, residual chlorine, water temperature and THM formation. MATERIALS AND METHODS: Drinking water samples were collected from different sampling points in the water treatment plant (WTP) from Gilau and the corresponding distribution system in Cluj-Napoca and also from Beclean, Dej and Jibou WTPs. The water samples were collected once a month from July 2006 to November 2007 and stored in 40-mL vials closed with Teflon lined screw caps. Water samples were preserved at 4 degrees C until analysis after sodium thiosulfate (Na(2)S(2)O(3)) had been added to quench residual chlorine. All samples were analysed for THMs using headspace GC-ECD between 1 and 7 days after sampling. The sample (10 mL) was filled into 20-mL headspace vials and closed with a Teflon-lined screw cap. Thereafter, the samples were equilibrated in an oven at 60 degrees C for 45 min. The headspace (1 mL) was then injected into the GC (Cyanopropylphenyl Polysiloxane column, 30 m x 53 mm, 3 microm film thickness, Thermo Finnigan, USA). The MDLs for THMs were determined from the standard deviation of eight standards at 1 microg/L. The MDLs for CHCl(3), CHBrCl(2), CHBr(2)Cl and CHBr(3) were 0.3, 0.2, 0.3 and 0.6 microg/L, respectively. All kinetic laboratory studies were carried out only with water from the WTP Gilau. The experiments were conducted under two conditions: baseline conditions (pH 7, 21 degrees C, 2.5 mg/L Cl(2)) to gain information about the change of the organic matter in the raw water and seasonally variable conditions to simulate the actual process at the treatment plant and the distribution system. RESULTS AND DISCUSSION: This study shows that the current chlorination practice in the investigated plants complies with the THM drinking water standards of the EU. The THM concentrations in all samples taken in the four treatment plants and distributions systems were below the EU drinking water standard for TTHMs of 100 microg/L. Due to the low bromide levels in the raw waters, the main THM formed in the investigated plants is chloroform. It could also be seen that the THM levels were typically lower in water supplies with groundwater as their water resource. In one plant (Dej) with a pre-ozonation step, a significantly lower (50%) THM formation during post chlorination was observed. Laboratory chlorination experiments revealed a good correlation between chloroform formation and the consumed chlorine dose. Also, these experiments allowed a semi-quantative prediction of the chloroform formation in the distribution system of Cluj-Napoca. CONCLUSIONS: CHCl(3) was the most important trihalomethane species observed after the chlorination of water in all of the sampled months. However, TTHM concentrations did not exceed the maximum permissible value of 100 microg/L (EU). The THM formation rates in the distribution system of Cluj-Napoca have a high seasonal variability. Kinetic laboratory experiments could be used to predict chloroform formation in the Cluj Napoca distribution system. Furthermore, an empirical model allowed an estimation of the chloroform formation in the Gilau water treatment plant. PMID- 19219475 TI - [Endosonography for differential diagnosis of malignant findings of the pancreas]. AB - Due to recent technical innovations endosonography has attained a special importance for the examination of malignant pancreatic tumors. Modern high resolution radial instruments with 360 degrees imaging give a very good overall view with fine resolution and contrast and allow excellent detection of solid space-occupying lesions in the region of the pancreas. In meta-analyses a very good sensitivity for tumor detection of over 90% could be demonstrated, comparable to the results of the modern CT pancreas spiral technique. Endosonography has an advantage over CT examination especially for the diagnostics of very small neuroendocrine tumors (<1 cm), for example for the detection of insulinomas. For cystic pancreas processes only endosonography allows further specific differential diagnostics by the determination of important prognostic tumor markers, such as CEA or CA-19-9 by simultaneous puncture of the lesion. PMID- 19219476 TI - [PET and PET-CT of malignant tumors of the exocrine pancreas]. AB - Adenocarcinomas of the pancreas represent the majority (>95%) of all malignant pancreatic tumors. They are formed from malignant degeneration of the exocrine part of the pancreas. Cystic acinar tumors are much rarer and originate from secretion-producing parenchymal cells of the pancreas. Endocrine tumors of the pancreas will not be dealt with in this context. PMID- 19219478 TI - Comment on Przybelski et al.: Rapid correction of low vitamin D status in nursing home residents. PMID- 19219479 TI - An open study of pamidronate in the treatment of refractory degenerative lumbar spinal stenosis. AB - Degenerative lumbar spinal stenosis, manifesting as chronic low back pain and neurogenic claudication, is an increasing chronic problem in an aging population, with limited effective conservative treatment options. Based on previous reports on the utility of subcutaneous calcitonin and two anectodal cases, we launched an open therapeutic trial of IV monthly pamidronate infusions, over a course of 3-6 months in this condition. Of 24 patients, 75% reported pain improvement, with the mean VAS score improved by 40%; while composite functional improvement in walking time, activities of daily living, and sense of well being was reported by 66%, with a mean improvement of 50%. The results of this pilot trial suggest the usefulness of this modality and warrant examination in a controlled clinical trial. PMID- 19219480 TI - Nailfold capillaroscopy in Behcet's disease, analysis of 128 patients. AB - The aims of this study were to find the characteristics and prevalence of nailfold capillary changes in a large series of patients with Behcet's disease (BD) and to analyze their possible relation to other clinical characteristics of the disease. We performed nailfold capillaroscopy in 128 randomly selected patients fulfilling the international classification criteria for BD. Capillaroscopy was done in eight fingers with a x3.2 microscopy. All patients were questioned for history of Raynaud's phenomenon, ischemic ulcers, smoking, and hypertension. A computerized form including demographic, clinical, and para clinical features was used to collect data. Univariate and multivariate logistic regressions were used to analyze the relation between capillaroscopic findings and disease characteristics. Odds ratio and a confidence interval at 95% (CI) were calculated for each item. The mean age of the patients was 37 +/- 10 years, and the male to female ratio was 1.56:1. Capillaroscopy was abnormal in 51 patients (40%, CI 8.5). Enlarged capillaries were seen in 33 patients (26%, CI 7.6), hemorrhages in 21 (16%, CI 6.4), and capillary loss only in one patient. In univariate logistic regression analysis, the presence of enlarged capillaries was associated with lower age at disease onset (OR = 0.9, CI 0.9-1; p = 0.04), hypertension (OR = 4.2, CI 1.5-11.4; p = 0.006), superficial phlebitis (OR = 5.5, CI 1.2-24.4; p = 0.03), and negative pathergy test (OR = 0.4, CI 0.2-0.9; p = 0.04). The presence of hemorrhages tended to be associated with articular symptoms (p = 0.05). Multivariate analysis also confirmed the association of enlarged capillaries with lower age at disease onset (p = 0.01), hypertension (p = 0.001), and superficial phlebitis (p = 0.03). Nailfold abnormalities, mainly enlarged capillaries, are frequent in patients with BD. Our results suggest that these abnormalities may be related to other vascular features of the disease such as superficial phlebitis, but it does not seem to confer special risk for any other specific clinical symptom of the disease. PMID- 19219481 TI - What factors determine patients' preference for tumour necrosis factor inhibitors in ankylosing spondylitis? AB - Tumour necrosis factor inhibitor (TNFi) therapy, either intravenous (IV) or subcutaneous (SQ), demonstrates similar efficacy in ankylosing spondylitis (AS). The objective of this study was to examine factors influencing patient preference of TNFi. Fifty-nine (79.7%) participants were male with mean age 43.9 years and disease duration of 22.0 years. Fifty-nine patients (79.7%) agreed with the statement 'My doctor gave me a choice and I made a decision based on my personal preference'. Patients commenced first on IV TNFi most commonly cited reduced frequency of injections (96.6%), administration by a trained professional (89.7%) and use of infusion time for leisure activities (86.2%). Patients commenced on SQ TNFi cited flexibility with timing of treatment (80%), shortened administration time (73.3%) and the convenience of home therapy (73.3%). Shared clinical decision-making between clinicians and patients may be desirable for AS patients commencing TNFi therapy. PMID- 19219482 TI - The improvement of middle ear ventilation by laser ablation of the epipharyngeal eustachian tube: a prospective study. AB - A long-lasting dysfunction of the eustachian tube seems to be the etiologic origin for development of chronic otitis media (COM) with mesotympanic perforation, otitis media with effusion (OME), and chronic atelectasis of the middle ear. Surgical interventions in the middle ear generally treat the sequelae of the tube dysfunction but not the dysfunction itself. This prospective clinical study investigated how far fiber-guided laser ablation of the posterior half of the epipharyngeal tubal ostium led to better middle ear ventilation in the otologic disease patterns mentioned below. There were 38 adult patients included in the analysis. The patients in one group had a perforated tympanic membrane [COM before primary tympanoplasty (n = 14) or revision tympanoplasty (n = 5)]; the patients in a second group had an intact eardrum [OME resistant to therapy (n = 3), with an atelectasis of the middle ear (n = 2) or problems of pressure equalization with fast changes in ambient pressure (diving, flying) (n = 14)]. Laser ablation of the posterior half of the epipharyngeal tubal ostium was performed, generally with local anesthesia, if tubal function testing was pathologic (Valsalva maneuver, passive tube opening, tympanogram). In patients with COM the procedure was performed 8 weeks before the middle ear surgery. All patients were checked 8 weeks postoperatively and in the course of the following year. The intervention seemed to have had a positive effect on tube function in 68.4% of patients operated on (P = 0.001). In 26 of the 38 patients that had undergone operation, an improvement the results of tubal function tests could be seen in the postoperative follow-up. In the COM group the Valsalva maneuver improved in 14 of 19 patients (73.6%) (P = 0.001), and the passive tube opening improved in nine of 18 patients (50%). In the group with an intact eardrum the Valsalva maneuver improved in 13 of 18 patients (72.2%) (P = 0.001). The resulting condition remained stable after 1 year. None of the patients showed any complications as a result of the therapy. Minimally invasive shaping of the distal eustachian tube under topical anesthesia can be recommended for patients with the above-mentioned diagnoses who have pathologic middle ear ventilation. Especially prior to tympanoplasties, and especially in otologic revision procedures, where middle ear ventilation is a prerequisite for successful otologic surgery, the function of the eustachian tube can be optimized in 70% of the patients, particularly if there are pathological findings (tubal tonsil, narrow orifice of the tubal ostium, adenoids). The placement of permanent ear tubes in adults with recurrent OME can also be avoided by the procedure described. The resultant conditions remained stable for the next year. Patients with tympanic ventilation problems due to rapid pressure changes (flying, diving) can also benefit from this procedure. PMID- 19219483 TI - Erbium, chromium:yttrium-scandium-gallium-garnet laser-assisted sinus graft procedure. AB - The possibility of using lasers in the field of dentistry has been a subject of investigation. There are few reports that any laser systems have been used for bony window osteotomy by direct sinus grafting. In this study, erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser of various laser systems was used for 12 sinus bone grafts in ten patients, and the efficiency of the laser was evaluated according to the osteotomy time and the rate of sinus membrane perforation in the clinical results; the mechanism is described. Eight of the 12 procedures were performed by direct sinus grafting with the Er,Cr:YSGG laser without membrane perforation (perforation ratio 33.3%). Operating time for bony window osteotomy with laser alone was 2-7 minutes [3 min 24 s on average; 3.4 +/- 1.4 min (mean +/- standard deviation)], and all the implants placed immediately were successful. PMID- 19219485 TI - Heat conduction from the exceedingly hot fiber tip contributes to the endovenous laser ablation of varicose veins. AB - Lower-extremity venous insufficiency is a common condition, associated with considerable health care costs. Endovenous laser ablation is increasingly used as therapy, but its mechanism of action is insufficiently understood. Here, direct absorption of the laser light, collapsing steam bubbles and direct fiber-wall contact have all been mentioned as contributing mechanisms. Because fiber tips have reported temperatures of 800-1,300 degrees C during endovenous laser ablation, we sought to assess whether heat conduction from the hot tip could cause irreversible thermal injury to the venous wall. We approximated the hot fiber tip as a sphere with diameter equal to the fiber diameter, having a steady state temperature of 800 degrees C or 1,000 degrees C. We computed venous wall temperatures due to heat conduction from this hot sphere, varying the pullback velocity of the fiber and the diameter of the vein. Venous wall temperatures corresponding to irreversible injury resulted for a 3 mm diameter vein and pullback velocities <3 mm/s but not for 5 mm and > or =1 mm/s. The highest wall temperature corresponded to the position on the wall closest to the fiber tip, hence it moves longitudinally in parallel with the moving fiber tip. We concluded that heat conduction from the hot fiber tip is a contributing mechanism in endovenous laser ablation. PMID- 19219484 TI - Effects of root planing procedures with hand instrument or erbium, chromium:yttrium-scandium-gallium-garnet laser irradiation on the root surfaces: a comparative scanning electron microscopy study. AB - The purpose of this study was to investigate the efficiency of hand instrumentation and laser irradiation on calculus removal from the root surfaces, in vitro. Thirty-two human teeth, extracted for periodontal reasons, were used in this study. Root surfaces of single-rooted teeth were treated by different methods including (1) conventional hand instruments; (2) hand instruments and tetracycline-hydrochloride (Tet-HCl); (3) erbium, chromium:yttrium-scandium gallium-garnet (Er,Cr:YSGG) laser irradiation, setting I (short pulse); (4) Er,Cr:YSGG laser irradiation, setting II (long pulse). Three premolar teeth, extracted for orthodontic reasons, served as control. The morphology of the root surfaces was evaluated by light and scanning electron microscopy. Energy dispersive X-ray (EDX) analysis was performed to compare the mineral content of root surfaces treated with hand instrumentation and lasing procedures. The results of this study demonstrated that all treatments were efficient in calculus removal from the root surfaces. Thermal changes, including melting and carbonization, were not observed in either lasing procedure. The surface was rougher in the laser groups than in the groups treated with hand instruments. Moreover, roughness was greater in the long-pulse laser setting than in the short pulse setting. While increased calcium (Ca) and decreased phosphate (P) (weight concentration percent) were observed in all treatments when compared with the control, laser procedures resulted in a more similar mineral content than in the groups treated with hand instruments. Based on these findings, laser procedures, when used in appropriate settings, are capable of performing scaling and root planing in the treatment of periodontitis. It may be concluded that short pulse laser may be more suitable for the micro-morphology of the root surface. However, additional in vitro and clinical studies are necessary to clarify the success of laser in periodontal therapy. PMID- 19219486 TI - Optimal head position during transesophageal echocardiographic probe insertion for pediatric patients weighing up to 10 kg. AB - No data exists for the optimal head position during transesophageal echocardiography (TEE) probe insertion in children. Turning the head to the side closes the ipsilateral pyriform sinus and dilates the contralateral pyriform sinus, theoretically making probe insertion safer and easier. In this study, 94 children (weight, 5 kg), II (weight, 4-5 kg), and III (weight, <4 kg). The difference was statistically significant only for subgroup III (p = 0.0001). Insertion failed in four children with the head in the midline position but was successful when the head was turned leftward. Children undergoing TEE who weigh less than 10 kg should have the head positioned to the side rather than in the midline position during probe insertion. If insertion is unsuccessful or difficult with the head in the midline position, the authors recommend turning the head to the side and reattempting probe insertion. PMID- 19219487 TI - Evolutional and geographical relationships of Bartonella grahamii isolates from wild rodents by multi-locus sequencing analysis. AB - To clarify the relationship between Bartonella grahamii strains and both the rodent host species and the geographic location of the rodent habitat, we have investigated 31 B. grahamii strains from ten rodent host species from Asia (Japan and China), North America (Canada and the USA), and Europe (Russia and the UK). On the basis of multi-locus sequencing analysis of 16S rRNA, ftsZ, gltA, groEL, ribC, and rpoB, the strains were classified into two large groups, an Asian group and an American/European group. In addition, the strains examined were clearly clustered according to the geographic locations where the rodents had been captured. In the phylogenetic analysis based on gltA, the Japanese strains were divided into two subgroups: one close to strains from China, and the other related to strains from Far Eastern Russia. Thus, these observations suggest that the B. grahamii strains distributed in Japanese rodents originated from two different geographic regions. In the American/European group, B. grahamii from the North American continent showed an ancestral lineage and strict host specificity; by contrast, European strains showed low host specificity. The phylogenetic analysis and host specificity of B. grahamii raise the possibility that B. grahamii strains originating in the North American continent were distributed to European countries by adapting to various rodent hosts. PMID- 19219488 TI - Identifying conservation and restoration priorities for saproxylic and old-growth forest species: a case study in Switzerland. AB - Saproxylic (dead-wood-associated) and old-growth species are among the most threatened species in European forest ecosystems, as they are susceptible to intensive forest management. Identifying areas with particular relevant features of biodiversity is of prime concern when developing species conservation and habitat restoration strategies and in optimizing resource investments. We present an approach to identify regional conservation and restoration priorities even if knowledge on species distribution is weak, such as for saproxylic and old-growth species in Switzerland. Habitat suitability maps were modeled for an expert-based selection of 55 focal species, using an ecological niche factor analyses (ENFA). All the maps were then overlaid, in order to identify potential species' hotspots for different species groups of the 55 focal species (e.g., birds, fungi, red listed species). We found that hotspots for various species groups did not correspond. Our results indicate that an approach based on "richness hotspots" may fail to conserve specific species groups. We hence recommend defining a biodiversity conservation strategy prior to implementing conservation/restoration efforts in specific regions. The conservation priority setting of the five biogeographical regions in Switzerland, however, did not differ when different hotspot definitions were applied. This observation emphasizes that the chosen method is robust. Since the ENFA needs only presence data, this species prediction method seems to be useful for any situation where the species distribution is poorly known and/or absence data are lacking. In order to identify priorities for either conservation or restoration efforts, we recommend a method based on presence data only, because absence data may reflect factors unrelated to species presence. PMID- 19219489 TI - Effective protection of open space: does planning matter? AB - High quality plans are considered a crucial part of good land use planning and often used as a proxy measure for success in plan implementation and goal attainment. We explored the relationship of open space plan quality to the implementation of open space plans and attainment of open space protection goals in Research Triangle, North Carolina, USA. To measure plan quality, we used a standard plan evaluation matrix that we modified to focus on open space plans. We evaluated all open space plans in the region that contained a natural resource protection element. To measure plan implementation and open space protection, we developed an online survey and administered it to open space planners charged with implementing the plans. The survey elicited each planner's perspective on aspects of open space protection in his or her organization. The empirical results (1) indicate that success in implementation and attaining goals are not related to plan quality, (2) highlight the importance of when and how stakeholders are involved in planning and implementation processes, and (3) raise questions about the relationship of planning to implementation. These results suggest that a technically excellent plan does not guarantee the long-term relationships among local land owners, political and appointed officials, and other organizations that are crucial to meeting land protection goals. A greater balance of attention to the entire decision process and building relationships might lead to more success in protecting open space. PMID- 19219490 TI - Evaluating aggregate terrestrial impacts of road construction projects for advanced regional mitigation. AB - This study presents a GIS-based database framework used to assess aggregate terrestrial habitat impacts from multiple highway construction projects in California, USA. Transportation planners need such impact assessment tools to effectively address additive biological mitigation obligations. Such assessments can reduce costly delays due to protracted environmental review. This project incorporated the best available statewide natural resource data into early project planning and preliminary environmental assessments for single and multiple highway construction projects, and provides an assessment of the 10-year state-wide mitigation obligations for the California Department of Transportation. Incorporation of these assessments will facilitate early and more strategic identification of mitigation opportunities, for single-project and regional mitigation efforts. The data architecture format uses eight spatial scales: six nested watersheds, counties, and transportation planning districts, which were intersected. This resulted in 8058 map planning units statewide, which were used to summarize all subsequent analyses. Range maps and georeferenced locations of federally and state-listed plants and animals and a 55-class landcover map were spatially intersected with the planning units and the buffered spatial footprint of 967 funded projects. Projected impacts were summarized and output to the database. Queries written in the database can sum expected impacts and provide summaries by individual construction project, or by watershed, county, transportation district or highway. The data architecture allows easy incorporation of new information and results in a tool usable without GIS by a wide variety of agency biologists and planners. The data architecture format would be useful for other types of regional planning. PMID- 19219491 TI - Framework and tools for agricultural landscape assessment relating to water quality protection. AB - While many scientific studies show the influence of agricultural landscape patterns on water cycle and water quality, only a few of these have proposed scientifically based and operational methods to improve water management. Territ'eau is a framework developed to adapt agricultural landscapes to water quality protection, using components such as farmers' fields, seminatural areas, and human infrastructures, which can act as sources, sinks, or buffers on water quality. This framework allows us to delimit active areas contributing to water quality, defined by the following three characteristics: (i) the dominant hydrological processes and their flow pathways, (ii) the characteristics of each considered pollutant, and (iii) the main landscape features. These areas are delineated by analyzing the flow connectivity from the stream to the croplands, by assessing the buffer functions of seminatural areas according to their flow pathways. Hence, this framework allows us to identify functional seminatural areas in terms of water quality and assess their limits and functions; it helps in proposing different approaches for changing agricultural landscape, acting on agricultural practices or systems, and/or conserving or rebuilding seminatural areas in controversial landscapes. Finally, it allows us to objectivize the functions of the landscape components, for adapting these components to new environmental constraints. PMID- 19219492 TI - Smoking knowledge, attitudes, behavior, and associated factors among Chinese male surgeons. AB - BACKGROUND: The purpose of this study was to understand and assess the smoking knowledge, attitudes, behavior, and associated factors among Chinese male surgeons. METHODS: A total of 823 Chinese male surgeons from six cities in China participated in a survey of smoking knowledge, attitudes, and behaviors in 2004. This study presents descriptive statistics and logistic regression analyses of factors associated with the respondents' smoking behavior and smoking cessation activity. RESULTS: The current smoking prevalence for Chinese male surgeons was 45.2, and 42.5% of respondents reported having smoked in front of their patients. Few of the respondents always asked patients about their smoking status (25%) or advised smokers to quit smoking (27.2%). Logistic regression models found that current smoking status was significantly associated (P < 0.05) with the surgeons' knowledge of the harms of active and passive smoking and their attitudes toward smoke-free hospitals and health role modeling by physicians. Smoking in front of patients was significantly associated (P < 0.05) with the respondents' knowledge of active smoking harms, attitudes toward smoke-free hospitals, and cigarette consumption. The surgeons' smoking cessation activity was significantly associated (P < 0.05) with their knowledge about the harms of active smoking, their rates of advising patients to quit smoking, and their knowledge of the harms of passive smoking. CONCLUSIONS: Male surgeons have the highest smoking prevalence among Chinese physicians. They should actively participate in tobacco control training and education to improve their knowledge and attitudes toward smoking, which will improve their own smoking behavior and smoking cessation practices. Only by engaging all parts of the health care system, including surgeons, can China make headway against its tobacco epidemic. PMID- 19219493 TI - Mortality impact of hypothermia after cavitary explorations in trauma. AB - BACKGROUND: Admission hypothermia (core temperature35 degrees C). The profoundly hypothermic subgroup of patients with a Tc<33 degrees C was also analyzed. RESULTS: During the study period, 2,489 patients required cavitary surgery, 1,252 of whom (50%) were admitted to the intensive care unit (ICU). On arrival in the ICU 15% of the patients had a Tcor=55 years old (12% versus 7%; p=0.04); in addition, they were hypotensive on admission (25% versus 13%; p<0.001), had a lower admission Glasgow Coma Score (GCS; 11+/-5 versus 14+/-3; p<0.001), a higher Injury Severity Score (ISS; 29+/-15 versus 22+/-12; p<0.001), higher head and chest Abbreviated Injury Scale (AIS), and greater intraoperative blood loss (2.6+/-2.4 l versus 1.7+/-1.8 l; p<0.001). When compared to patients who were normothermic at the end of their operation, hypothermic patients had a significantly higher mortality (35% versus 8%; p<0.001). With decreasing Tc, there was a stepwise increase in mortality. Compared to patients with a Tc>35 degrees C, the relative risk of death for patients with a Tc between 35 degrees C and 33 degrees C was 4.0, and that for patients with a Tcor=0.5 mg/l). These results confirmed that the combination of SHV-1, CTX-M-3, CTX-M-14, TEM-1, or OXA-11 production and reduced expression of ompK35/36 may not result in clinical resistance to imipenem but does correlate with increasing imipenem MIC. PMID- 19219498 TI - Molecular cloning and expression of a novel cholinephosphotransferase involved in glycoglycerophospholipid biosynthesis of Mycoplasma fermentans. AB - A gene, mf1, encoding a novel cholinephosphotransferase in glycoglycerophospholipid (GGPL) biosynthesis of Mycoplasma fermentans PG18 was identified by genomic analysis, cloned, and expressed in Escherichia coli. The mf1 gene comprises an open reading frame of 777 bp encoding 258 amino acids. The mf1 gene product, Mf1, has 23% amino acid homology with LicD of Haemophilus influenzae but no homology with genes of other Mycoplasma species in the GenBank database. The reaction product of Mf1 using alpha-glucopyranosyl-1,2 dipalmitoilglycerol and cytidine 5'-diphosphocholine (CDP-choline) as substrates showed the specific protonated molecule at m/z 896, which corresponded to GGPL-I as determined by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS). Furthermore, the product ions of choline, phosphocholine, and hexose-bound phosphocholine were detected by tandem mass spectrometry (MS) analysis of protonated molecules at m/z 896. These results identified mf1 as a novel cholinephosphotransferase and showed that the phosphocholine transfer step is involved in the GGPL biosynthesis pathway of M. fermentans. This is the first report of a GGPL biosynthesis enzyme. PMID- 19219499 TI - Metabolic shift in the phytopathogen Rhodococcus fascians in response to cell free extract of infected tobacco plant tissues. AB - The phytopathogen Rhodococcus fascians induces the development of leafy gall, which is considered to be its ecological niche. To obtain a view of the metabolic changes occurring in R. fascians during this process, an in vitro system was used where bacteria are grown in the presence of a leafy gall extract, a condition mimicking that found by the bacteria in infected plants. Proteins of R. fascians grown for 24 h under these conditions were displayed by two-dimensional polyacrylamide gel electrophoresis. Fifteen polypeptides showing a differential accumulation in response to the inducing conditions were analyzed by mass spectrometry. Two polypeptides potentially linked to the Krebs cycle, a pyruvate dehydrogenase and a fumarate hydratase, were further characterized and shown to be downregulated at the transcriptional level. The identification of these two enzymes suggests that R. fascians may shift its metabolism during the interaction with plants from the Krebs cycle to the glyoxylate shunt. PMID- 19219500 TI - Antimicrobial susceptibility profiles of Salmonella enterica serotypes recovered from pens of commercial feedlot cattle using different types of composite samples. AB - Salmonella enterica in cattle production systems may be associated with important human and animal disease issues. However, tremendous diversity exists among Salmonella recovered, and more information is needed about strains of greatest potential health concern, particularly those that are multidrug resistant (MDR). By characterizing Salmonella isolates from commercial feedlot pens, this study aimed to evaluate the strain diversity and prevalence of MDR Salmonella from different types of composite pen samples. Antimicrobial susceptibility profiles, serotype, and presence or absence of the integron-encoded intI1 gene were determined for 530 Salmonella isolates recovered using composite rope (n = 335), feces (n = 59), and water (n = 136) samples from 21 pens in 3 feedlots. The study investigated only pens with available isolates from multiple sample types. Most isolates (83.0%) of the 19 Salmonella serotypes identified were susceptible or intermediately susceptible to all the antimicrobials evaluated. Resistance to sulfisoxazole (14.9%), streptomycin (3.8%), and tetracycline (3.6%) were the most common. None of the isolates tested positive for a class 1 integron, and only 2.5% were resistant to multiple antimicrobials. All the MDR isolates, namely, serotypes Uganda (n = 9), Typhimurium (n = 2), and Give (n = 2), were resistant to at least five antimicrobials. Most MDR isolates (n = 11) were from two pens during 1 week within one feedlot. Overall, many Salmonella isolates collected within a pen were similar in terms of serotype and antimicrobial susceptibility regardless of sample type. However, MDR Salmonella and rare serotypes were not recovered frequently enough to suggest a general strategy for appropriate composite sampling of feedlot cattle populations for Salmonella detection and monitoring. PMID- 19219501 TI - The bovine dilated cardiomyopathy locus maps to a 1.0-Mb interval on chromosome 18. AB - Cardiomyopathies are myocardial diseases that lead to cardiac dysfunction, heart failure, arrhythmia, and sudden death. In human medicine, cardiomyopathies frequently warrant heart transplantation in children and adults. Bovine dilated cardiomyopathy (BDCMP) is a heart muscle disorder that has been observed during the last 30 years in cattle of Holstein-Friesian origin. In Switzerland BDCMP affects Swiss Fleckvieh and Red Holstein breeds. BDCMP is characterized by a cardiac enlargement with ventricular remodeling and chamber dilatation. The common symptoms in affected animals are subacute subcutaneous edema, congestion of the jugular veins, and tachycardia with gallop rhythm. A cardiomegaly with dilatation and hypertrophy of all heart chambers, myocardial degeneration, and fibrosis are typical postmortem findings. It was shown that all BDCMP cases reported worldwide traced back to a red factor-carrying Holstein-Friesian bull, ABC Reflection Sovereign. An autosomal recessive mode of inheritance was proposed for BDCMP. Recently, the disease locus was mapped to a 6.7-Mb interval MSBDCMP06 BMS2785 on bovine Chr 18 (BTA18). In the present study the BDCMP locus was fine mapped by using a combined strategy of homozygosity mapping and association study. A BAC contig of 2.9 Mb encompassing the crucial interval was constructed to establish the correct marker order on BTA18. We show that the disease locus is located in a gene-rich interval of 1.0 Mb and is flanked by the microsatellite markers DIK3006 and MSBDCMP51. PMID- 19219502 TI - Chronic sleep complaints in premenopausal women and their association with sleep disordered breathing. AB - BACKGROUND: In clinical practice, we have found that premenopausal women have delayed diagnosis of sleep-disordered breathing (SDB). METHODS: During a 4-year period, we systematically collected the clinical and polysomnographic variables for all women referred for sleep complaints using preestablished questionnaires, scales, clinical grid, polygraphic montage, and scoring criteria. The variables collected on premenopausal SDB women were analyzed and compared to those of postmenopausal women within 5 years of menopause. RESULTS: Of 977 women, 316 were premenopausal with SDB. Complaints of chronic insomnia and sleepwalking were the most common reasons for referral, had been present for a mean of 6.4 +/- 5.4 years, and had lead to unsuccessful symptomatic treatment. The normal-weight premenopausal SDB group had anatomically small upper airways, while those with body mass index (BMI) >/= 25 kg/m(2) complained more frequently of snoring and daytime sleepiness and their clinical presentation was closer to those of the postmenopausal SDB comparison group. Premenopausal women often had a low apnea hypopnea index (AHI), but there was a discrepancy between the low AHI and the amount of continuous positive airway pressure (CPAP) needed to control the SDB, and there was a need for higher pressures in overweight premenopausal SDB women (mean 9.1 +/- 1.9 and 10.1 +/- 2.6 cmH(2)O). CONCLUSION: Normal-weight premenopausal SDB women often present with atypical sleep complaints of chronic insomnia and parasomnias. Clinical attention paid to craniofacial features and use of specific scales such as Mallampati help with the suspicion of the presence of SDB, and a low AHI is unrelated to the positive clinical impact of nasal CPAP treatment. PMID- 19219503 TI - HLA antigens and nontuberculous mycobacterial lung disease in Korean patients. AB - BACKGROUND: Human leukocyte antigen (HLA) molecules are known to play an important role in host-defense mechanisms. The aim of this study was to evaluate the association between HLA alleles and lung disease caused by nontuberculous mycobacteria (NTM) in Korean patients. METHODS: Seventy-eight patients with NTM lung disease (48 patients with Mycobacterium avium-intracellulare complex [MAC] infection and 30 patients with Mycobacterium abscessus infection) were included in the study. HLA-A, -B, and -DRB1 genotyping was performed by polymerase chain reaction using sequence-specific primers. Data from 485 healthy Korean individuals were used as a control. RESULTS: When compared to controls, patients with NTM lung disease showed an increased frequency of DRB1*11 (OR = 1.91, 95% confidence interval [CI] = 1.01-3.64, p = 0.045, corrected p [pC] > 0.05). In the subgroup analysis, patients with MAC lung disease had an increased frequency of B*46 (OR = 2.23, 95% CI = 1.05-4.73, p = 0.044, pC > 0.05). CONCLUSIONS: Our data suggest that in a Korean population, patients with NTM lung disease and healthy subjects differ in the frequencies of some HLA alleles. However, when considering corrected p values, our findings are inconclusive. PMID- 19219504 TI - Automatic segmentation of the ribs, the vertebral column, and the spinal canal in pediatric computed tomographic images. AB - We propose methods to perform automatic identification of the rib structure, the vertebral column, and the spinal canal in computed tomographic (CT) images of pediatric patients. The segmentation processes for the rib structure and the vertebral column are initiated using multilevel thresholding and the results are refined using morphological image processing techniques with features based on radiological and anatomical prior knowledge. The Hough transform for the detection of circles is applied to a cropped edge map that includes the thoracic vertebral structure. The centers of the detected circles are used to derive the information required for the opening-by-reconstruction algorithm used to segment the spinal canal. The methods were tested on 39 CT exams of 13 patients; the results of segmentation of the vertebral column and the spinal canal were assessed quantitatively and qualitatively by comparing with segmentation performed independently by a radiologist. Using 13 CT exams of six patients, including a total of 458 slices with the vertebra from different sections of the vertebral column, the average Hausdorff distance was determined to be 3.2 mm with a standard deviation (SD) of 2.4 mm; the average mean distance to the closest point (MDCP) was 0.7 mm with SD = 0.6 mm. Quantitative analysis was also performed for the segmented spinal canal with three CT exams of three patients, including 21 slices with the spinal canal from different sections of the vertebral column; the average Hausdorff distance was 1.6 mm with SD = 0.5 mm, and the average MDCP was 0.6 mm with SD = 0.1 mm. PMID- 19219505 TI - Decrease in hemoglobin levels following surgery influences the outcome in head and neck cancer patients treated with accelerated postoperative radiotherapy. AB - AIM: To assess the influence of hemoglobin (Hb) levels in locally advanced head and neck cancer (LAHNC) patients treated with surgery and postoperative radiotherapy (PORT). MATERIAL AND METHODS: Pre- and postoperative Hb levels were collected in 79 patients treated with surgery followed by accelerated PORT for LAHNC. Median follow-up was 52 months (range 12-95 months). RESULTS AND DISCUSSION: Four-year overall survival (OS) rate was 51%. Neither pre- nor postoperative Hb level (<120 or 130 g/l in women or men, respectively) influenced the outcome. However, when Hb decrease between pre- and postoperative Hb values was taken into account, 4-year OS was significantly higher in patients with Hb difference less than 38 g/l (quartile value) compared with those with Hb decrease 38 g/l or more (61% versus 16%, P = 0.008). CONCLUSION: Decrease in Hb level by more than 38 g/l after surgery secondary to blood loss influences the outcome when postoperative RT is indicated. PMID- 19219506 TI - Accuracy of endoscopic ultrasound to diagnose nodal invasion by rectal cancers: a meta-analysis and systematic review. AB - BACKGROUND: Nodal staging in patients with rectal cancer predicts prognosis and directs therapy. Published data on the accuracy of endoscopic ultrasound (EUS) for diagnosing nodal invasion in patients with rectal cancer has been inconsistent. AIM: To evaluate the accuracy of EUS in diagnosing nodal metastasis of rectal cancers. METHOD: Study Selection Criteria: Only EUS studies confirmed by surgical histology were selected. Data Collection and Extraction: Articles were searched in Medline, Pubmed, and CENTRAL. STATISTICAL METHOD: Pooling was conducted by both fixed-effects model and random-effects model. RESULTS: The initial search identified 3610 reference articles in which 352 relevant articles were selected and reviewed. Data were extracted from 35 studies (N = 2732) that met the inclusion criteria. Pooled sensitivity of EUS in diagnosing nodal involvement by rectal cancers was 73.2% (95% confidence interval [95% CI] 70.6 75.6). EUS had a pooled specificity of 75.8% (95% CI 73.5-78.0). The positive likelihood ratio of EUS was 2.84 (95% CI 2.16-3.72), and negative likelihood ratio was 0.42 (95% CI 0.33-0.52). All the pooled estimates, calculated by fixed- and random-effect models, were similar. SROC curves showed an area under the curve of 0.79. The P for chi-squared heterogeneity for all the pooled accuracy estimates was >.10. CONCLUSIONS: EUS is an important and accurate diagnostic tool for evaluating nodal metastasis of rectal cancers. This meta-analysis shows that the sensitivity and specificity of EUS is moderate. Further refinement in EUS technologies and diagnostic criteria are needed to improve the diagnostic accuracy. PMID- 19219507 TI - Ki-67 expression gives additional prognostic information on St. Gallen 2007 and Adjuvant! Online risk categories in early breast cancer. AB - BACKGROUND: We sought to determine the significance of Ki-67, one of the tumor cell proliferation markers, as a useful prognostic factor in early breast cancer. METHODS: A total of 1080 consecutive patients with stage I or II breast cancer that underwent surgery between 1998 and 2003 were enrolled. Patients were categorized on the basis of the 2007 St. Gallen consensus and Adjuvant! Online. The expression of Ki-67 in the tumor was assayed by immunohistochemistry (cutoff value, 10%). RESULTS: Univariate analysis determined that tumor size, lymph node involvement, histologic grade, estrogen receptor, progesterone receptor, bcl-2, and Ki-67 (> or =10%) were statistically significant for both overall survival (OS) and distant metastasis-free survival (DFS). Of these factors, lymph node involvement and high Ki-67 expression were identified as independent prognostic factors for OS and DFS on the basis of multivariate analysis. The survivals of intermediate- and high-risk groups according to 2007 St. Gallen consensus were further separated by Ki-67 expression level (5-year DFS rate = 91.9% vs. 86.3% for Ki-67 < 10% and > or = 10%, respectively in intermediate-risk group (P = .01); 5-year DFS rate = 82.5% vs. 61.4% for Ki-67 < 10% and > or = 10%, respectively in high-risk group (P = .01)). The survivals of low- and high-risk groups according to Adjuvant! Online were further separated by Ki-67 expression level (5-year DFS rate = 97.8% vs. 89.5% for Ki-67 < 10% and > or = 10%, respectively in low-risk group (P = .02); 5-year DFS rate = 9.4% vs. 82.6% for Ki 67 < 10% and > or = 10% in high-risk group (P = .005)). CONCLUSIONS: Ki-67 is an independent prognostic factor for DFS and OS in early breast cancer and can provide additional prognostic information on the risk stratification with the use of the 2007 St. Gallen consensus and Adjuvant! Online. PMID- 19219508 TI - Stroke volume variation as a predictor of intravascular volume depression and possible hypotension during the early postoperative period after esophagectomy. AB - BACKGROUND: Perioperative hypotension during esophagectomy results from hypovolemia caused by a shift of extracellular fluid from the intravascular to the extravascular compartment. Fluid management is often difficult to gauge during major surgery because there are no reliable indicators of fluid status, and some patients still experience cardiorespiratory instability. In this retrospective study, we evaluated stroke volume variation (SVV), calculated by using a new arterial pressure-based cardiac output measurement device, as a predictor for fluid responsiveness after esophageal surgery. METHODS: Eighteen patients undergoing esophagectomy with extended radical lymphadenectomy were monitored by the FloTrac sensor/Vigileo monitor system during the perioperative and immediate postoperative period. Fluid responsiveness was assessed and compared with concurrent SVV and central venous pressure (CVP) values, and routine hemodynamic variables. RESULTS: Eleven of 18 patients needed additional volume loading within the first 10 postoperative hours as a result of hypotension. The maximum SVV value of fluid resuscitated patients was >15% in all cases, whereas six of seven patients without postoperative hypotension had maximum SVV values of <15%. The correlation between SVV and the development of hypotension was statistically significant (P = 0.0012). From the linear correlation analysis of hemodynamic variables influenced by additional fluid loading, SVV was significantly correlated to cardiac output (r = 0.638; P = 0.049), whereas CVP was not (P > 0.05). CONCLUSION: We conclude that SVV, as displayed on the Vigileo monitor, is an accurate predictor of intravascular hypovolemia and is a useful indicator for assessing the appropriateness and timing of applying fluid for improving circulatory stability during the perioperative period after esophagectomy. PMID- 19219509 TI - Ecological invasion, roughened fronts, and a competitor's extreme advance: integrating stochastic spatial-growth models. AB - Both community ecology and conservation biology seek further understanding of factors governing the advance of an invasive species. We model biological invasion as an individual-based, stochastic process on a two-dimensional landscape. An ecologically superior invader and a resident species compete for space preemptively. Our general model includes the basic contact process and a variant of the Eden model as special cases. We employ the concept of a "roughened" front to quantify effects of discreteness and stochasticity on invasion; we emphasize the probability distribution of the front-runner's relative position. That is, we analyze the location of the most advanced invader as the extreme deviation about the front's mean position. We find that a class of models with different assumptions about neighborhood interactions exhibits universal characteristics. That is, key features of the invasion dynamics span a class of models, independently of locally detailed demographic rules. Our results integrate theories of invasive spatial growth and generate novel hypotheses linking habitat or landscape size (length of the invading front) to invasion velocity, and to the relative position of the most advanced invader. PMID- 19219510 TI - Continuous probabilistic analysis to evolutionary game dynamics in finite populations. AB - Evolutionary game dynamics of two strategies in finite population is studied by continuous probabilistic approach. Besides frequency dependent selection, mutation was also included in this study. The equilibrium probability density functions of abundance, expected time to extinction or fixation were derived and their numerical solutions are calculated as illustrations. Meanwhile, individual based computer simulations are also done. A comparison reveals the consistency between theoretical analysis and simulations. PMID- 19219511 TI - Dynamic modeling of herpes simplex virus type-2 (HSV-2) transmission: issues in structural uncertainty. AB - The sexually transmitted infection (STI) Herpes simplex virus type-2 (HSV-2) is of public health concern because it is a very common frequently unrecognized lifelong infection, which may facilitate HIV transmission. Within HIV/STI modeling, structural uncertainty has received less attention than parametric uncertainty. By merging the compartments of a "complex" model, a "simple" HSV-2 model is developed. Sexual interactions between female sex workers (FSWs) and clients are modeled using data from India. Latin Hypercube Sampling selects from parameter distributions and both models are run for each of the 10,000 parameter sets generated. Outputs are compared (except for 2,450 unrealistic simulations). The simple model is a good approximation to the complex model once the HSV-2 epidemic has reached 60% of the equilibrium prevalence (95% of the 7,550 runs produced <10% relative error). The simple model is a reduced version of the complex model that retains details implicitly. For late-stage epidemics, the simple model gives similar prevalence trends to the complex model. As HSV-2 epidemics in many populations are advanced, the simple model is accurate in most instances, although the complex model may be preferable for early epidemics. The analysis highlights the issue of structural uncertainty and the value of reducing complexity. PMID- 19219513 TI - A foundation of failure. PMID- 19219512 TI - Patient-provider communication about sexual health among unmarried middle-aged and older women. AB - BACKGROUND: Although past studies have highlighted the importance of patient provider communication about sexual health and intimate relationships (SHIR), much of the research has focused on young women's or married women's experiences when discussing SHIR with their providers. OBJECTIVE: To describe experiences of unmarried, middle-aged and older women in communicating about SHIR with their health care providers. DESIGN AND PARTICIPANTS: Qualitative interviews were conducted with 40 unmarried women aged 40-75 years. We compared the responses of 19 sexual minority (lesbian and bisexual) women and 21 heterosexual women. RESULTS: Women varied in their definitions of intimate relationships. Not all women thought providers should ask about SHIR unless questions were directly related to a health problem, and most were not satisfied with questions about SHIR on medical intake forms. However, the themes women considered to be important in communication about SHIR were remarkably consistent across subgroups (e.g. previously married or never married; sexual minority or heterosexual). Sexual minority women were more hesitant to share information about SHIR because they had had prior negative experiences when disclosing their sexual orientation or perceived that clinicians were not informed about relevant issues. CONCLUSIONS: Some women felt that providers should ask about SHIR only if questions relate to an associated health problem (e.g. sexually transmitted infection). When providers do ask questions about SHIR, they should do so in ways that can be answered by all women regardless of partnering status, and follow questions with non-judgmental discussions. PMID- 19219514 TI - Gallstone formation after gastric cancer surgery. AB - BACKGROUND: Gallstone formation is one of the most common complications after gastric cancer surgery, but the mechanism and etiology for such formation are unclear because of a lack of collective clinical investigation. METHOD: We evaluated the influence of various surgical factors on the incidence of gallstone formation after gastrectomy. Gallstone formation was confirmed by ultrasound examinations that were routinely carried out after surgery on a periodic basis. RESULTS: Gallstone formation occurred in 173 of 672 (25.7%) patients who had undergone gastrectomy with lymph-node dissection for gastric cancer. The types of gastrectomy and reconstruction had no significant effect on the incidence, but the extent of lymph-node dissection was a significant factor (p < 0.001: D1 + alpha vs. D2 + alpha; p < 0.01: D2 vs. D2 + alpha). Gallstones were usually formed within 2 years after gastrectomy, but in most cases, gallstone formation was asymptomatic. CONCLUSION: The extent of lymph-node dissection was a significant factor in gallstone formation after gastrectomy; therefore, prophylactic cholecystectomy should be considered in cases of extensive lymph node dissection. PMID- 19219515 TI - What to do when the pathology from last week's laparoscopic cholecystectomy is malignant and T1 or T2. PMID- 19219516 TI - Treatment of war wounds: a historical review. AB - The treatment of war wounds is an ancient art, constantly refined to reflect improvements in weapons technology, transportation, antiseptic practices, and surgical techniques. Throughout most of the history of warfare, more soldiers died from disease than combat wounds, and misconceptions regarding the best timing and mode of treatment for injuries often resulted in more harm than good. Since the 19th century, mortality from war wounds steadily decreased as surgeons on all sides of conflicts developed systems for rapidly moving the wounded from the battlefield to frontline hospitals where surgical care is delivered. We review the most important trends in US and Western military trauma management over two centuries, including the shift from primary to delayed closure in wound management, refinement of amputation techniques, advances in evacuation philosophy and technology, the development of antiseptic practices, and the use of antibiotics. We also discuss how the lessons of history are reflected in contemporary US practices in Iraq and Afghanistan. PMID- 19219517 TI - Long-term periprosthetic remodeling in THA shows structural preservation. AB - Although periprosthetic changes after THA have been well documented in short-term studies of less than 5 years, little is known about long-term changes. Long-term mineral changes must be evaluated against an unaffected limb control and for regional differences about a prosthesis. This study evaluated long-term periprosthetic remodeling using dual-energy x ray absorptiometry in a prospective study of patients who had noncemented THAs with a modular titanium alloy proximal loading prosthesis. In 15 randomly selected patients, bone mineral content was measured within 15 months of surgery and then at late mean followup of 13 years. In the affected femur, there was a major decrease in periprosthetic bone mineral content in Zones 1, 2, 6, and 7 (Gruen et al.) over the course of the study. The overall decrease in Zone 7 was 39% in bone mineral content. Estimates made after controlling for the contralateral unaffected femur indicate a major loss only in Zone 7 and preservation of mineral content in Zones 3, 4, and 5 of the proximal femur. The data suggest bone remodeling maintains the overall structural integrity of the upper femoral shaft. PMID- 19219518 TI - The classic: observations on pathogenesis and treatment of congenital clubfoot. 1972. PMID- 19219519 TI - The classic: congenital club foot: the results of treatment. 1963. PMID- 19219520 TI - Prevalence of coronary artery disease across the Framingham risk categories: coronary artery calcium scoring and MSCT coronary angiography. AB - BACKGROUND: Non-invasive assessment of subclinical atherosclerosis by means of coronary artery calcium scoring (CACS) and multi-slice computed tomography (MSCT) coronary angiography could improve patients' risk stratification. However, data relating observations on CACS and MSCT coronary angiography to traditional risk assessment are scarce. METHODS AND RESULTS: In 314 consecutive outpatients (54 +/ 13 years, 56% males) without known CAD, CACS and 64-slice MSCT coronary angiography were performed. According to the Framingham risk score (FRS), 51% of patients were at low, 24% at intermediate and 25% at high risk, respectively. MSCT angiograms showing atherosclerosis were classified as showing obstructive (> or =50% luminal narrowing) CAD or not. Both CACS and MSCT coronary angiography showed a high prevalence of normal coronary arteries in low FRS patients (70% and 61%, respectively). An increase in the prevalence of CACS >400 (4% low vs 19% intermediate vs 36% high), CAD (39% low vs 79% intermediate vs 91% high), and obstructive CAD (15% low vs 43% intermediate vs 58% high) was observed across the FRS categories (P < .0001 for all comparisons). CONCLUSIONS: A strong positive relationship exists between FRS and the prevalence and extent of atherosclerosis. Especially in intermediate FRS patients, CACS and MSCT coronary angiography provide useful information on the presence of subclinical atherosclerosis. PMID- 19219521 TI - Nuclear cardiology needs new "blood". PMID- 19219522 TI - Seventh annual Mario S. Verani, MD Memorial Lecture: "ASNC and integrating cardiac imaging". AB - Although cardiovascular mortality is decreasing over the last two decades, the cost of such care is escalating at a pace that is not sustainable for Medicare. Over this same time period, cardiac imaging has grown at an extremely rapid rate and is a leading contributor to this escalation in expense. It is now incumbent on both profession medical societies and individual physicians to demonstrate that cardiovascular care and the imaging growth lead to better patient outcomes and that the level of competent performance in these areas is documented. The development of physician certification and laboratory accreditation are key components to this effort and need to be supported and updated in a timely manner. The recent introduction of appropriateness criteria for patient imaging and treatment, especially in concert with the more established national clinical guidelines, appears to offer the best mechanism to deal with all the clinical and economic factors that we will need to address to continue our efforts to improve cardiovascular healthcare. Physicians can best serve their patients by practicing best standards of practice as well as being advocates for high-quality medical care in the setting of responsible healthcare funding. PMID- 19219523 TI - Microbial transformation and bioactivation of isoflavones from Pueraria flowers by human intestinal bacterial strains. AB - The flowers from Pueraria, which are called Puerariae Flos, have been used since ancient times for recovery from alcohol intoxication. We elucidated the microbial transformation of the main isoflavones (1, 1a and 2) by using 29 commercially available human intestinal bacterial strains together with the bioactivation of the hepatoprotective activity of their metabolites. Tectoridin (1a), which contains only one glucosyl moiety, was metabolized to its aglycone 1b by various bacterial strains. On the other hand, the metabolism of 1 and 2, which both have disaccharide groups, was limited to specific bacterial strains. The metabolites 1c and 2c obtained from the Peptostreptococcus productus strain were completely different from those produced by the other strains. These metabolites were identified as 6-hydroxygenistein and 6-hydroxybiochanin A, respectively. The glycosides 1, 1a and 2 did not show any hepatoprotective activity, whereas aglycones 1b and 2b showed moderate activity. Furthermore, the hepatoprotective activity of the demethylated metabolites 1c and 2c was extremely potent. Although not all people have P. productus in their gastrointestinal tract, the O demethylated compounds might become one of the bioactivated metabolites when Puerariae Flos is administered orally. PMID- 19219524 TI - Chemical constituents of the whole plants of Saussurea medusa. AB - The chemical constituents of the traditional Tibetan medicine of Saussurea medusa Maxim. (Compositae) were investigated and a new flavonoid glucoside, together with 14 known compounds, was isolated. The structure of the new compound was established as 6''-O-crotonoylhomoplantaginin by using one- and two-dimensional nuclear magnetic resonance spectroscopy and mass spectrometry analyses. PMID- 19219525 TI - Peroxiredoxins: a less studied component of hydrogen peroxide detoxification in photosynthetic organisms. AB - Peroxiredoxins (Prx) are ubiquitous thiol-dependent peroxidases capable of reducing a broad range of toxic peroxides and peroxinitrites. A cysteinyl residue of peroxiredoxins reacts with the peroxides as primary catalytic center and oxidizes to sulfenic acid. The regeneration of the reduced form of Prx is required as a next step to allow its entry into next catalytic cycle. Several proteins, such as thioredoxin, glutaredoxin, cyclophilin, among others, are known to facilitate the regeneration of the reduced (catalytically active) form of Prx in plants. Based on the cysteine residues conserved in the deduced amino acid sequence and their catalytic mechanisms, four groups of peroxiredoxins have been distinguished in plants, namely, 1-Cys Prx, 2-Cys Prx, Type II Prx and Prx Q. Peroxiredoxins are known to play an important role in combating the reactive oxygen species generated at the level of electron transport activities in the plant exposed to different types of biotic and abiotic stresses. In addition to their role in antioxidant defense mechanisms in plants, they also modulate redox signaling during development and adaptation. Besides these general properties, peroxiredoxins have been shown to protect DNA from damage in vitro and in vivo. They also regulate metabolism in thylakoids and mitochondria. The present review summarizes the most updated information on the structure and catalysis of Prx and their functional importance in plant metabolism. PMID- 19219526 TI - The use and misuse of journal metrics and other citation indicators. AB - This article reviews the nature and use of the journal impact factor and other common bibliometric measures for assessing research in the sciences and social sciences based on data compiled by Thomson Reuters. Journal impact factors are frequently misused to assess the influence of individual papers and authors, but such uses were never intended. Thomson Reuters also employs other measures of journal influence, which are contrasted with the impact factor. Finally, the author comments on the proper use of citation data in general, often as a supplement to peer review. This review may help government policymakers, university administrators, and individual researchers become better acquainted with the potential benefits and limitations of bibliometrics in the evaluation of research. PMID- 19219528 TI - Rank-normalized journal impact factor as a predictive tool. AB - Citation data accumulated on articles from the top and bottom 25 of impact factor (IF)-ranked international journals are compared using 59 international geoscience journals from 1998 and 378 Polish geological papers from 1989-1994. There is a minor risk of being un-cited when results are published in high-IF periodicals as the average non-citation rate is 0.88 over a 10-year period in this not very rapidly developing scientific discipline. Similarly, the established error levels in the prognosis of expected citation success versus failure based on the extreme IF quartiles as an evaluation tool is low (at most 12.5). Thus the application of the rank-normalized journal IF as a proxy of real citation frequency and, accordingly, as a predictive tool in the a priori qualification of recently published publications is a rational time- and cost-saving alternative (or at least a significant supplement) to traditional informed peer review. Blanket criticism of using IF for decisions in research funding is therefore at least partly exaggerated. PMID- 19219529 TI - Biological activity of dendritic cells generated from cord blood CD34+ hematopoietic progenitors in IL-7- and IL-13-conditioned cultures. AB - INTRODUCTION: Dendritic cells (DCs) are required for initiation of the immune response and may therefore be used for the production of cancer vaccines. As mature DCs (mDCs) are the most potent antigen-presenting cells, there is increasing interest in generating them ex vivo. The present study was designed to obtain mDCs from CD34+ hematopoietic progenitors by culturing them in different media. MATERIALS AND METHODS: Cord blood CD34+ hematopoietic progenitors were expanded for 7 days in FST medium containing fms-related tyrosine kinase 3 ligand (Flt3-L), stem cell factor (SCF), and thrombopoietin (TPO). Then the cells were divided into three parts and cultured for 21 days in different media: FST medium or FST enriched in interleukin (IL)-3 (FST3 medium) or supplemented with IL-7 and IL-13 (FST713 medium). At the end of culture part of the cells was harvested, counted, and analyzed while the other part was matured with proinflammatory cytokines for 2 days. The cells' phenotypes, ability to induce proliferation of allogeneic lymphocytes in the mixed lymphocyte reaction (allo-MLR), chemotaxis, phagocytosis, and O2(-) production were determined. RESULTS: The average fold increase of DCs at the end of culture in FST medium was 127, in FST3 1043, and in FST713 71. In comparison with the other media, FST713 medium supported the generation of mDCs that were characterized by higher expression of CD83, costimulatory molecules, and HLA-DR, enhanced ability to induce allo-MLR and migration to -macrophage inflammatory protein (MIP) 3beta poor phagocytosis, and O2(-) production. CONCLUSIONS: This study indicates that FST713 medium allows the generation of limited numbers of more mature DCs, while FST3 medium leads to the production of immature DCs in high numbers. PMID- 19219530 TI - The use of bibliometric indicators to help peer-review assessment. AB - Inserm is the only French public research institution entirely dedicated to human health. Inserm supports research across the biomedical spectrum in all major disease areas, from fundamental lab-based science to clinical trials. To translate its scientists' findings into tangible health benefits, Inserm has its own affiliated company, Inserm Transfert, which works with industry. Since 2001, Inserm has been setting up on-line file management software for the evaluation of researchers and laboratories, called EVA ( www.eva.inserm.fr ). EVA includes all grant applications, assessment reports, evaluation grading evaluation forms and includes automated bibliometric indicator software that enables calculating, for example, the number of publications, journal impact factors, number of citations, citation index, and number of the Top 1 publications for each researcher of the teams. The indicators take into account research fields, the year of publications, and the author's position among the participants. Bibliometrics is now considered a tool for science policy providing indicators to measure productivity and scientific quality, thereby supplying a basis for evaluating and orienting R&D. It is also a potential tool for evaluation. It is neutral, allows comparative (national and international) assessment, and may select papers in the forefront in all fields. For each team, bibliometric indicators are calculated for all researchers with permanent or long-term positions (3-5 years). The use of bibliometric indicators requires great vigilance, but according to our experience they enrich the committee's debates without any doubt. We present an analysis of the data of 600 research teams evaluated in 2007-2008. PMID- 19219531 TI - The use of bibliometrics to measure research quality in UK higher education institutions. AB - Research assessment in the UK has evolved over a quarter of a century from a loosely structured, peer-review based process to one with a well understood data portfolio and assessment methodology. After 2008, the assessment process will shift again, to the use of indicators based largely on publication and citation data. These indicators will in part follow the format introduced in 2008, with a profiling of assessment outcomes at national and international levels. However, the shift from peer assessment to a quantitative methodology raises critical issues about which metrics are appropriate and informative and how such metrics should be managed to produce weighting factors for funding formulae. The link between publication metrics and other perceptions of research quality needs to be thoroughly tested and reviewed, and may be variable between disciplines. Many of the indicators that drop out of publication data are poorly linked to quality and should not be used at all. There are also issues about which publications are the correct base for assessment, which staff should be included in a review, how subjects should be structured and how the citation data should be normalised to account for discipline-dependent variables. Finally, it is vital to consider the effect that any assessment process will have on the behaviour of those to be assessed. PMID- 19219527 TI - Virulence factor genotypes of Helicobacter pylori affect cure rates of eradication therapy. AB - The cure rates of Helicobacter pylori infection by using a combination of a proton pump inhibitor (PPI) and antimicrobial agents are mainly influenced by bacterial susceptibility to antimicrobial agents and the magnitude of acid inhibition during the treatment. Currently used empirical triple therapies do not reliably produce a > or =80% cure rate on an intention-to-treat basis. Therefore, tailored regimens based on relevant microbiological findings and pharmacogenomics are recommended for attaining an acceptable > or =95% cure rate. Recently, virulence factors of H. pylori, such as cagA and vacA, are reported to be major factors determining the cure rates. Individuals infected with strains with cagA negative and vacA s2 genotypes have significantly increased risk of eradication failure of H. pylori infection. These virulence factors enhance gastric mucosal inflammation and are associated with the development of peptic ulcer and gastric cancer. H. pylori virulence factors induce proinflammatory cytokines, such as interleukin (IL)-1, IL-8, and tumor necrosis factor (TNF)- which influence mucosal inflammation and/or gastric acid secretion. When physicians select an H. pylori eradication regimen with an acceptable cure rate, they might need to consider H. pylori virulence factors, especially cagA and vacA. PMID- 19219533 TI - New developments in the use of citation analysis in research evaluation. AB - This paper presents an overview of research assessment methodologies developed in the field of evaluative bibliometrics, a subfield of quantitative science and technology studies, aimed to construct indicators of research performance from a quantitative statistical analysis of scientific-scholarly documents. Citation analysis is one of its key methodologies. The paper illustrates the potentialities and limitations of the use of bibliometric indicators in research assessment. It discusses the relationship between metrics and peer review; databases used as sources of bibliometric analysis; the pros and cons of indicators often applied, including journal impact factors, Hirsch indices, and normalized indicators of citation impact; and approaches to the bibliometric measurement of institutional research performance. PMID- 19219532 TI - Sjogren's syndrome: an old tale with a new twist. AB - Sjogren's syndrome (SjS) is chronic autoimmune disease manifested by the loss of saliva and/or tear secretion by salivary and/or lacrimal glands, respectively. The pathogenesis of the disease remains elusive, perhaps due to the multiple triggers of the disease. However, substantial advances have been made in attempting to resolve the complexity of SjS using both animal models and human subjects. The primary objectives of this review are to provide a better understanding of the disease processes with major emphasis on the use of mouse models, how genetic predisposition plays a role in the natural history of the disease, as well as a presentation of new findings pertaining to the role of T(H)1, T(H)2, and T(H)17 cells in the pathogenesis of SjS. PMID- 19219534 TI - The role of the calponin homology domain of smoothelin-like 1 (SMTNL1) in myosin phosphatase inhibition and smooth muscle contraction. AB - In this study, we provide further insight into the contribution of the smoothelin like 1 (SMTNL1) calponin homology (CH)-domain on myosin light chain phosphatase (SMPP-1M) activity and smooth muscle contraction. SMTNL1 protein was shown to have inhibitory effects on SMPP-1M activity but not on myosin light chain kinase (MLCK) activity. Treatment of beta-escin permeabilized rabbit, ileal smooth muscle with SMTNL1 had no effect on the time required to reach half-maximal force (t(1/2)) during stimulation with pCa6.3 solution. The addition of recombinant SMTNL1 protein to permeabilized, smooth muscle strips caused a significant decrease in contractile force. While the calponin homology (CH)-domain was essential for maximal SMTNL1-associated relaxation, it alone did not cause significant changes in force. SMTNL1 was poorly dephosphorylated by PP-1C in the presence of the myosin targeting subunit (MYPT1), suggesting that phosphorylated SMTNL1 does not possess "substrate trapping" properties. Moreover, while full length SMTNL1 could suppress SMPP-1M activity toward LC(20) in vitro, truncated SMTNL1 lacking the CH-domain was ineffective. In summary, our findings suggest an important role for the CH-domain in mediating the effects of SMTNL1 on smooth muscle contraction. PMID- 19219535 TI - Association of genetic variants in Methylenetetrahydrofolate Reductase and Paraoxonase-1 genes with homocysteine, folate and vitamin B12 in coronary artery disease. AB - BACKGROUND: The aim of the present study was to investigate the association between genetic variants in metylenetetrahydrofolate reductase (MTHFR) and Paraoxonase-1 (PON1) 55/192 genes and total homocysteine (tHcy), folate, B12 vitamin, and PON1 levels in patients with coronary artery disease (CAD). METHODS: The study included 235 patients with CAD and 268 healthy control subjects. RESULTS: LL and LM genotypes and L allele of PON1 55 were over-represented in patients. In contrast, MM genotype and M allele were more frequent in controls. QQ genotype and Q allele of PON1 192 and CT genotype of MTHFR were significantly diminished and QR genotype and R allele were significantly elevated in CAD patients compared with controls. The plasma tHcy were elevated but B12 levels were diminished in patients. PON1 55 and 192 genetic variants were significantly associated with PON1 activity, triglyceride, total cholesterol, tHcy and, high density lipoprotein-cholesterol and low-density lipoprotein-cholesterol in patients, respectively. CONCLUSION: Genetic variants of PON1 55/192 and MTHFR were associated with CAD. PMID- 19219536 TI - Physiotherapists and use of low back pain guidelines: a qualitative study of the barriers and facilitators. AB - INTRODUCTION: A new set of clinical practice guidelines (CPGs) for the management of low back pain (LBP) and prevention of persistent disability entitled "Clinic on Low back pain in Interdisciplinary Practice" (CLIP) was developed in the province of Quebec, Canada. The literature shows that simply disseminating CPGs does not necessarily lead to their adoption by clinicians. To improve adherence to CPGs among healthcare professionals, the literature suggests that there is a need to identify and address the factors impeding or facilitating their use. The aim of this study was therefore to identify the barriers to and facilitators of CLIP CPG use, as perceived by physiotherapists (PTs). METHODS: A descriptive study using a qualitative method was conducted with a sample of 16 PTs from a variety of professional backgrounds. Each participant used the CPGs over a 6-week period with two patients suffering from LBP, and then participated in a semi structured interview in which he or she was asked to identify the barriers and facilitators experienced. RESULTS: The participating PTs identified many barriers and facilitators pertaining to the guidelines themselves, the users and the environment. Four key nodes emerged from these barriers and facilitators during data analysis. It appears that the clinicians' understanding of the CPGs, the level of compatibility between their practices and the CLIP CPG recommendations, the level of CPG relevance as perceived by the clinicians, and their level of agreement with the CPGs, all affected their use of the guidelines. CONCLUSIONS: In order to increase CLIP CPG use, the implementation strategy to be developed should take into account the barriers and facilitators that were identified in this study. PMID- 19219537 TI - Exercise for the primary, secondary and tertiary prevention of low back pain in the workplace: a systematic review. AB - INTRODUCTION: Low back pain (LBP) is one of the most costly conditions to manage in occupational health. Individuals with chronic or recurring LBP experience difficulties returning to work due to disability. Given the personal and financial cost of LBP, there is a need for effective interventions aimed at preventing LBP in the workplace. The aim of this systematic review was to examine the effectiveness of exercises in decreasing LBP incidence, LBP intensity and the impact of LBP and disability. METHODS: A comprehensive literature search of controlled trials published between 1978 and 2007 was conducted and a total of 15 studies were subsequently reviewed and analyzed. RESULTS: There was strong evidence that exercise was effective in reducing the severity and activity interference from LBP. However, due to the poor methodological quality of studies and conflicting results, there was only limited evidence supporting the use of exercise to prevent LBP episodes in the workplace. Other methodological limitations such as differing combinations of exercise, study populations, participant presentation, workloads and outcome measures; levels of exercise adherence and a lack of reporting on effect sizes, adverse effects, and types of sub-groups, make it difficult to draw definitive conclusions on the efficacy of workplace exercise in preventing LBP. CONCLUSIONS: Only two out of the 15 studies reviewed were high in methodological quality and showed significant reductions in LBP intensity with exercise. Future research is needed to clarify which exercises are effective and the dose-response relationships regarding exercise and outcomes. PMID- 19219538 TI - The impact of misspecification of residual error or correlation structure on the type I error rate for covariate inclusion. AB - It has been shown that when using the FOCE method in NONMEM, the likelihood ratio test (LRT) can be sensitive to the use of an inappropriate estimation method in that ignoring an existing eta-epsilon interaction leads to actual significance levels for type I errors being higher than the nominal levels. The objective of this study was to assess through simulations the LRT sensitivity to various types of residual error model misspecifications in both continuous and categorical data. The study contained two parts, simulations based on continuous and categorical data. Data sets containing 250 individuals with up to 24 observations per individual were simulated multiple times (1000) with different types of residual error models for the continuous data and different strength of correlation between observations for the categorical data. The data sets were analyzed using either the correct or a simpler (incorrect) model with or without addition of a covariate. The type I error rate of inclusion of the non informative covariate on the 5% level was calculated as the number of runs where the drop in the objective function value (OFV) was larger than 3.84 when the covariate relationship was included in the model using the correct or the incorrect model. The difference in OFV between the model with the correct and the incorrect structure was also calculated as a measure of the residual error model misspecification. For continuous data the FOCE method was used in most cases (with interaction when appropriate). The Laplacian estimation method was used for one of the continuous models and for categorical data. The results showed that the residual error model misspecifications when the erroneous model was used were pronounced, as indicated by the OFV being substantially higher than for the corresponding correct models. The significance levels of the LRT with the incorrect model were appropriate in all cases but ignoring (serial) correlations between observations (continuous and categorical data) as well as when the eta epsilon interaction was ignored (which has previously been shown, continuous data). When ignoring correlation, the type I error rates were shown to be sensitive to the correlation strength, the number of observations per individual and the magnitude of the inter-individual variability on clearance. We conclude that the LRT appears robust towards all tested cases, but ignoring (serial) correlations between observations and eta-epsilon interaction. PMID- 19219539 TI - Plasma 1,25 dihydroxy vitamin D3 level and expression of vitamin d receptor and cathelicidin in pulmonary tuberculosis. AB - INTRODUCTION: Vitamin D(3), which exerts its effect through vitamin D receptor (VDR), is known for its potent immunomodulatory activities. Associations between low serum vitamin D(3) levels and increased risk of tuberculosis have been reported. STUDY SUBJECTS AND METHODS: Plasma 1,25 dihydroxy vitamin D(3) levels (1,25(OH)(2) D(3)) and ex vivo levels of VDR protein from peripheral blood mononuclear cells were studied in 65 pulmonary tuberculosis (PTB) patients and 60 normal healthy subjects (NHS) using enzyme-linked immunosorbent assay-based methods. Using real-time polymerase chain reaction (PCR), induction of VDR, cathelicidin, and CYP27B1 mRNA were studied in live Mycobacterium tuberculosis stimulated macrophage cultures treated with or without 1,25 dihydroxy vitamin D(3). VDR and CYP27B1 (-1077 A/T) gene polymorphisms were studied using PCR-based methods. RESULTS: 1,25(OH)(2) D(3) were significantly increased (p = 0.0004), while ex vivo levels of VDR protein were significantly decreased in PTB patients (p = 0.017) as compared to NHS. 1,25(OH)(2) D(3) levels were not different between variant genotypes of CYP27B1. A trend towards decreased levels of VDR protein was observed among NHS with BsmI BB and TaqI tt genotypes compared to NHS with other genotypes. Relative quantification of mRNA using real-time PCR revealed increased VDR mRNA expression in live M. tuberculosis-stimulated culture in PTB patients (p < 0.01) than normal healthy subjects. Cathelicidin mRNA expression was significantly increased in vitamin D(3)-treated cultures compared to unstimulated and M. tuberculosis-stimulated culture in both patients (p < 0.001) and NHS (p < 0.05). CONCLUSIONS: The present study suggests that PTB patients may have increased 1,25(OH)(2) D(3) levels, and this might lead to downregulation of VDR expression. Decreased VDR levels could result in defective VDR signaling. Moreover, addition of 1,25(OH)(2) D(3) might lead to increased expression of cathelicidin which could enhance the immunity against tuberculosis. PMID- 19219540 TI - Experiences and decisions that motivate women at increased risk of breast cancer to participate in an experimental screening program. AB - Although the discovery of mutations on BRCA1 and BRCA2 genes associated with high breast cancer risk has given rise to screening and surveillance initiatives, there is little documentation on why high-risk women choose to enter screening programs. The objective of this qualitative study was to develop a detailed understanding of the experiences and decisions that motivate women with increased risk of hereditary breast cancer to participate in the multicentered Quebec experimental breast screening program. Our study involved 21 participants who were either BRCA carriers or at risk and untested. These women were interviewed while participating in the screening program. Our study demonstrates that intensive screening programs may provide valuable reassurance for women with increased familial risk of hereditary breast cancer, who count on early detection and rapid response from professionals if and when a problem arises. Health professionals must take these and others concerns into account to ensure their interventions are most consistent with women's needs. PMID- 19219541 TI - The Psychological Inventory of Criminal Thinking Styles and Psychopathy Checklist: screening version as incrementally valid predictors of recidivism. AB - A follow-up of 107 male federal prison inmates previously tested with the Psychological Inventory of Criminal Thinking Styles (PICTS) and Psychopathy Checklist: Screening Version (PCL:SV) was conducted to test the incremental validity of both measures. The PICTS General Criminal Thinking (GCT) score was found to predict general recidivism and serious recidivism when age, prior charges, and the PCL:SV were controlled. The PCL:SV, on the other hand, failed to predict general and serious recidivism when age, prior charges, and the PICTS were controlled. These findings support the hypothesis that content-relevant self report measures like the PICTS are capable of predicting crime-relevant outcomes above and beyond the contributions of basic demographic variables like age, criminal history, and such popular non-self-report rating procedures as the PCL:SV. PMID- 19219542 TI - Tomato extract for hypertension? Editorial to "The effects of natural antioxidants from tomato extract in treated but uncontrolled hypertensive patients" by E. Paran et al. PMID- 19219543 TI - Patterns of sexual coercion in heterosexual and non-heterosexual men and women. AB - Heterosexual sex differences in evolved sexual interests, sexual scripts, and aggressive tendencies have each been hypothesized to be responsible for heterosexual sex differences in sexually coercive behavior. The current study compared the sexually coercive behavior of heterosexual and non-heterosexual men and women as a means of evaluating the explanatory capabilities of each of these hypotheses. A total of 414 participants were asked to provide information regarding their sexual coercion perpetration behavior since puberty by using a modified version of the Sexual Experiences Survey. Participants also completed measures of masculinity, femininity, and aggressive tendencies. Heterosexual men were significantly more likely to have perpetrated physical sexual coercion than all other participant groups. However, these group differences ceased to exist after controlling for group differences in physical aggression. Also, heterosexual men were significantly more likely to have perpetrated non-physical sexual coercion than all other groups, and non-heterosexual men were significantly more likely to have perpetrated non-physical sexual coercion than heterosexual and non-heterosexual women. Only the sexual orientation difference among men ceased to exist after controlling for masculinity or physical aggression. Discussion focused on detailing which hypotheses regarding heterosexual sex differences in sexually coercive behavior were supported as well as directions for future research. PMID- 19219544 TI - Hebephilia plethysmographica: a partial rejoinder to Blanchard et al. (2008). PMID- 19219545 TI - Relationship between mental disorders/suicidality and three sexual behaviors: results from the National Comorbidity Survey Replication. AB - The present study examined the relationship between sexual behaviors and mental disorders and suicidality in the National Comorbidity Survey Replication, a representative sample of adults ages 18 years and older (N = 5,692). The World Health Organization Composite International Diagnostic Interview was used to make DSM-IV based disorder diagnoses. Participants were also asked about suicidality and sexual behaviors. Multiple logistic regression analyses adjusted for sociodemographic variables were used to examine the relationships of three sexual behaviors (age of first intercourse, number of past year partners, and past year condom use) with 15 mental disorders (clustered into any mood, anxiety, substance use, and disruptive behavior groups) and suicidality (ideation and attempts). Compared to ages 15-17, those with age of first intercourse between 12 and 14 had increased rates of lifetime disruptive behavior, substance use, and any mental disorder, and suicidal ideation and attempts (adjusted odds ratio (AOR) range, 1.46-2.01). Those with age of first intercourse between ages 18-25 and 26-35 were at decreased likelihood of several lifetime disorder groups (AOR range, 0.19 0.81). Individuals who had two or more sexual partners in the past year had increased rates of all past year disorder groups examined (AOR range, 1.44-5.01). Never married participants who rarely/never used condoms were more likely than those who always used condoms to experience any mood, substance use, and any mental disorder, and suicide attempts (AOR range, 1.77-8.13). Future research should longitudinally examine these associations and account better for possible familial and personality confounders. PMID- 19219546 TI - Social policies in Uruguay: a view from the political dimension of community psychology. AB - This paper approaches social policy (SP) from the political perspective considered by Montero (Critical psychology: An introduction, Sage, London, pp 231 244, 1997; Community, Work and Family 1(1):65-78, 1998; Introduccion a la Psicologia Comunitaria. Desarrollo, conceptos y procesos, Paidos, Buenos Aires, pp 89-114, pp 255-284, 2004; Hacer para transformar. El metodo en la Psicologia Comunitaria, Paidos, Buenos Aires, pp 229-256, 2004) as a paradigmatic dimension of community psychology. The field of SP, the characteristics of certain SPs and the role of SP in the production and reproduction of the subjectivities of those subject to them in Uruguay are described and an analysis given of the challenge posed by going from a compensatory or palliative to a transformative mode of SP. It is proposed that identification of, and understanding and intervention in, psychosocial processes in the field are the keys to maximising the likelihood of SPs assuming a transformative character. Psychosocial aspects and participatory processes implicit in the relationship between the State and civil society are discussed. Finally, some orientations for community psychological intervention in this field of action are proposed. PMID- 19219547 TI - Structural characteristics and in vitro macrophage activation of acetyl fucoidan from Cladosiphon okamuranus. AB - We investigated a structural characteristics of acetyl fucoidan (CAF) isolated from commercially cultured Cladosiphon okamuranus. The CAF-induced macrophage activation and its signaling pathways in murine macrophage cell line, RAW 264.7 were also investigated. From the results of methylation analysis, CAF consisted of alpha-1-->3 linked L: -fucosyl residues and substituted sulfate and acetyl groups at C-4 on the main chain. CAF induced production of nitric oxide (NO), tumor necrosis factor-alpha and interleukin-6 in RAW 264.7 cells. Sulfate and acetyl groups of CAF involved in CAF-induced NO production. Neutralizing anti Toll-like receptor 4 (TLR4), anti-CD14 and anti-scavenger receptor class A (SRA) but not anti-complement receptor type 3 monoclonal antibodies decreased CAF induced NO production. The results of immunoblot analysis indicated that CAF activated mitogen-activated protein kinases (MAPKs) such as p38 MAPK, extracellular signal-regulated kinase (ERK)1/2 and stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK). SB203580 (p38 MAPK inhibitor) and SP600125 (SAPK/JNK inhibitor), but not U0126 (MAPK/ERK kinase 1/2 inhibitor) decreased CAF-induced NO production. The results suggested that CAF induced macrophage activation through membrane receptors TLR4, CD14 and SRA, and MAPK signaling pathways. PMID- 19219548 TI - VEGF recruits lactosylceramide to induce endothelial cell adhesion molecule expression and angiogenesis in vitro and in vivo. AB - Angiogenesis is largely driven by vascular endothelial growth factor (VEGF). However, the role of lipid second messengers such as lactosylceramide (LacCer) and LacCer synthase in angiogenesis is not well understood. We have determined the distribution of various LacCer synthase mRNA transcripts using sequential analysis of gene expression (SAGE). Endothelial cells from colon cancer tissues had a 4.5-fold increase in a LacCer synthase transcript (beta1,4GalT-V) as compared to normal colon tissue endothelial cells. Consequently, our focus turned to understanding the role of this enzyme in regulating VEGF-induced angiogenesis in vitro and in vivo. Herein, we show that in human endothelial cells, VEGF induced angiogenesis is mitigated by dimethylsphingosine and suramin; inhibitors of sphingosine kinase 1(SphK-1) and sphingosine1-phosphate receptor 1(S1P (1)), respectively, and this were bypassed by LacCer but not by S1P. VEGF and basic fibroblast growth factor-induced angiogenesis was mitigated by PDMP; an inhibitor of glucosylceramide synthase and LacCer synthase in human umbilical vein endothelial cells (HUVEC) and human aortic endothelial cells (HAEC). Likewise, GalT-V gene ablation using corresponding siRNA also mitigated VEGF-induced angiogenesis. In Matrigel plug angiogenesis assay in nude mice, angiogenesis was markedly inhibited by D-PDMP with concordantly diminished LacCer synthase activity. Mechanistic studies revealed that the use of LY294002, a PI3 kinase inhibitor, mitigated VEGF-induced expression of platelet-endothelial cell adhesion molecule (PECAM-1/CD31); the trans-endothelial migration of a monocyte cell line (U-937) and angiogenesis in HAEC cells. Since this enzyme is a target for VEGF action and LacCer serves as a lipid second messenger in inducing angiogenesis in vitro and in vivo, novel therapeutic approaches may be developed using our findings to mitigate colon cancer. PMID- 19219549 TI - Protective effect of resveratrol against kainate-induced temporal lobe epilepsy in rats. AB - Resveratrol (Res) is a phytoalexin produced naturally by several plants, which has multi functional effects such as neuroprotection, anti-inflammatory, and anti cancer. The present study was to evaluate a possible anti-epileptic effect of Res against kainate-induced temporal lobe epilepsy (TLE) in rat. We performed behavior monitoring, intracranial electroencepholography (IEEG) recording, histological analysis, and Western blotting to evaluate the anti-epilepsy effect of Res in kainate-induced epileptic rats. Res decreased the frequency of spontaneous seizures and inhibited the epileptiform discharges. Moreover, Res could protect neurons against kainate-induced neuronal cell death in CA1 and CA3a regions and depressed mossy fiber sprouting, which are general histological characteristics both in TLE patients and animal models. Western blot revealed that the expression level of kainate receptors (KARs) in hippocampus was reduced in Res-administrated rats compared to that in epileptic ones. These results suggest that Res is a potent anti-epilepsy agent, which protects against epileptogenesis and progression of the kainate-induced TLE animal. PMID- 19219550 TI - Effects of bicyclol on liver regeneration after partial hepatectomy in rats. AB - Bicyclol is a synthetic antihepatitis drug with antioxidative property. The present study was performed to investigate the effect of bicyclol on liver regeneration after partial hepatectomy in rats. Bicyclol (300 mg/kg) was given to rats subjected to 70% hepatectomy three times before operation. At 6, 24, and 48 h after resection, samples were collected for the measurement of serum alanine aminotransferase (ALT), total bilirubin (TBil), hepatic glycogen, malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione (GSH). Moreover, liver regeneration rate, proliferating cell nuclear antigen (PCNA) labeling, proliferation index, and histopathological examination were evaluated at 48 h after hepatectomy. As a result, bicyclol significantly increased regeneration rate, mitotic index (MI), PCNA labeling index, and proliferation index in PH rats. Additionally, bicyclol remarkably inhibited the elevation of serum ALT and TBil levels, alleviated the formation of liver MDA, restored impaired antioxidant SOD and GSH, increased hepatic glycogen content, and also attenuated hepatic vacuolar degeneration. These results suggested that bicyclol had a beneficial effect on liver regenerative capacity of the remnant liver tissue after hepatectomy, probably due to its antioxidative property. PMID- 19219551 TI - Imaging activity of neuronal populations with new long-wavelength voltage sensitive dyes. AB - We have assessed the utility of five new long-wavelength fluorescent voltage sensitive dyes (VSD) for imaging the activity of populations of neurons in mouse brain slices. Although all the five were capable of detecting activity resulting from activation of the Schaffer collateral-CA1 pyramidal cell synapse, they differed significantly in their properties, most notably in the signal-to-noise ratio of the changes in dye fluorescence associated with neuronal activity. Two of these dyes, Di-2-ANBDQPQ and Di-1-APEFEQPQ, should prove particularly useful for imaging activity in brain tissue and for combining VSD imaging with the control of neuronal activity via light-activated proteins such as channelrhodopsin-2 and halorhodopsin. PMID- 19219552 TI - Differences in mental health service sector utilization among African American and Caucasian youth entering systems of care programs. AB - Differences in unmet need and access to services between African American and Caucasian youth have been established; less is known about differences in specific mental health service sectors. This study examined differences in past year outpatient, school-based, day treatment and residential/inpatient service utilization among African American and Caucasian youth (n = 3,649) entering a federally funded system of care program. Random effect logistic regression models were implemented to examine the relationship between race and past year service utilization. Analyses revealed that African American youth were less likely than Caucasian youth to have utilized school-based and residential/inpatient mental health services in the past year. Findings suggest that racial disparities exist in service use for certain types of service sectors and highlight the importance of understanding and identifying individual, family, and community factors that contribute to disparities in service utilization. PMID- 19219553 TI - From genotype to phenotype: a longitudinal study of a patient with hypertrophic cardiomyopathy due to a mutation in the MYBPC3 gene. AB - Many of the links between the genotype and phenotype in hypertrophic cardiomyopathy remain unexplained. In this unique longitudinal study we have investigated a patient with classical clinical phenotypic features of hypertrophic obstructive cardiomyopathy, with a known mutation in MYBPC3, the most commonly affected gene in this disease. By collecting cardiac tissue from the patient at the time of surgical myectomy for relief of left ventricular outflow tract obstruction, we have been able to examine the structure of the myocytes and the functional differences that occur in MyBP-C mutated HCM cardiac tissue from single protein level, onto single cardiomyocyte contractility, through to whole organ function as assessed clinically by echocardiography. PMID- 19219554 TI - Myocardial adaptations in the failing heart: cause or consequence? AB - Many changes in morphology, biochemical properties and myocyte function occur during development to heart failure. Most changes may be compensatory, yet unable to prevent cardiac dysfunction in the long run. This illustrates that it is important to carefully dissect the disease causing modifications from cardiac adaptation, in order to obtain a better understanding of the pathophysiological processes leading to heart failure. PMID- 19219556 TI - An efficient synthesis of substituted 2-iminothiazolidin-4-one and thiadiazoloquinazolinone derivatives. AB - A convenient synthesis of functionalized 2-iminothiazolidin-4-ones and thiadiazoloquinazolinones has been achieved by the reaction of dialkyl acetylenedicarboxylates (DAAD) with 1-pheny-3-pyridin-2-yl-thiourea (1) and 3 amino-2-mercapto-3H-quinazolin-4-one (8), respectively. The starting materials for these processes were prepared from phenyl isothiocyanate, 2-aminopyridine for (1) and anthranilic acid, carbon disulfide, hydrazine for (8). Treatment of these compounds with DAAD resulted in the formation of an intermediate that readily underwent intramolecular cyclization to afford functionalized five membered heterocyclic rings was formed. PMID- 19219555 TI - Fibronectins in vascular morphogenesis. AB - Fibronectin is an extracellular matrix protein found only in vertebrate organisms containing endothelium-lined vasculature and is required for cardiovascular development in fish and mice. Fibronectin and its splice variants containing EIIIA and EIIIB domains are highly upregulated around newly developing vasculature during embryogenesis and in pathological conditions including atherosclerosis, cardiac hypertrophy, and tumorigenesis. However, their molecular roles in these processes are not entirely understood. We review genetic studies examining functions of fibronectin and its splice variants during embryonic cardiovascular development, and discuss potential roles of fibronectin in vascular disease and tumor angiogenesis. PMID- 19219557 TI - A novel quantitative structure-activity relationship model for prediction of biomagnification factor of some organochlorine pollutants. AB - The biomagnification factor (BMF) is an important property for toxicology and environmental chemistry. In this work, quantitative structure-activity relationship (QSAR) models were used for the prediction of BMF for a data set including 30 polychlorinated biphenyls and 12 organochlorine pollutants. This set was divided into training and prediction sets. The result of diversity test reveals that the structure of the training and test sets can represent those of the whole ones. After calculation and screening of a large number of molecular descriptors, the methods of stepwise multiple linear regression and genetic algorithm (GA) were used for the selection of most important and significant descriptors which were related to BMF. Then multiple linear regression and artificial neural network (ANN) techniques were applied as linear and non-linear feature mapping techniques, respectively. By comparison between statistical parameters of these methods it was concluded that an ANN model, which used GA selected descriptors, was superior over constructed models. Descriptors which were used by this model are: topographic electronic index, complementary information content, XY shadow/XY rectangle and difference between partial positively and negatively charge surface area. The standard errors for training and test sets of this model are 0.03 and 0.20, respectively. The degree of importance of each descriptor was evaluated by sensitivity analysis approach for the nonlinear model. A good results (Q (2) = 0.97 and SPRESS = 0.084) is obtained by applying cross-validation test that indicating the validation of descriptors in the obtained model in prediction of BMF for these compounds. PMID- 19219558 TI - Pseudo five-component synthesis of bis-alpha-acyloxy-beta-diketo amides from diimide-dicarboxylic acids. AB - Diimide-dicaboxylic acids derived from condensation of alpha-amino acids with the 2,3,5,6-bicyclo[2.2.2] oct-2-enetetracarboxylic-2,3:5,6-dianhydride or 1,2,4,5- bezenetetracarboxylic-1,2:4,5-dianhydride react with ninhydrin and isocyanides to produce some novel interesting bis-alpha-acyloxy-beta-diketo amides. PMID- 19219559 TI - Similarity-based SIBAR descriptors for classification of chemically diverse hERG blockers. AB - There is an increasing interest in computational models for the classification and prediction of the human ether-a-go-go-related-gene (hERG) potassium channel affinity in the early phase of drug discovery and development. In this study, similarity-based SIBAR descriptors were applied in order to develop and validate in silico binary QSAR and counter-propagation neural network models for the classification of hERG activity. The SIBAR descriptors were calculated based on four reference datasets using four sets of 2D- and 3D-descriptors including 3D grid-based VolSurf, 3D 'inductive' QSAR, Van der Waals surface area (P_VSA) and a set of 11 hERG relevant 2D descriptors devised from feature selection methods. The results indicate that the reference data set tailored to the specific problem, together with a set of hERG relevant descriptors, provides highly predictive models. PMID- 19219560 TI - Prediction of interaction between small molecule and enzyme using AdaBoost. AB - The knowledge of whether one enzyme can interact with a small molecule is essential for understanding the molecular and cellular functions of organisms. In this paper, we introduce a classifier to predict the small molecule- enzyme interaction, i.e., whether they can interact with each other. Small molecules are represented by their chemical functional groups, and enzymes are represented by their biochemical and physicochemical properties, resulting in a total of 160 features. These features are input into the AdaBoost classifier, which is known to have good generalization ability to predict interaction. As a result, the overall prediction accuracy, tested by tenfold cross-validation and independent sets, is 81.76% and 83.35%, respectively, suggesting that this strategy is effective. In this research, we typically choose interactions between small molecules and enzymes involved in metabolism to ultimately improve further understanding of metabolic pathways. An online predictor developed by this research is available at http://chemdata.shu.edu.cn/small_m . PMID- 19219561 TI - Characterization of human skin-derived mesenchymal stem cell proliferation rate in different growth conditions. AB - This study investigated conditions for optimal in vitro propagation of human skin derived mesenchymal stem cells (S-MSC). Forty primary skin-derived precursor cell (SKP) cultures were established from both male and female donors (age 29-65 years) and eight of them were randomly selected for in-depth characterization. Effects of basic fibroblast growth factor (FGF-2), epidermal growth factor (EGF), leukemia inhibiting factor (LIF) and dibutyryl-cyclic adenosine monophosphate (db cAMP) on S-MSC proliferation were investigated. Primary SKP cultures were >95% homogenous for CD90, CD73, and CD105 marker expression enabling to classify these cells as S-MSC. FGF-2 dose-dependent stimulation was observed in low serum medium only, whereas EGF neither stimulated S-MSC proliferation nor potentates the effect of FGF-2. Pronounced donor to donor differences among S-MSC cultures were observed in 3-day proliferation assay. This study demonstrates that homogenous S MSC populations can be reproducibly isolated from individual donors of different age. Optimal cell culture conditions for in vitro propagation of S-MSC are B27 supplemented or low serum media with FGF-2 (4 ng/ml). EGF and LIF as well as db cAMP are dispensable for S-MSC proliferation. PMID- 19219562 TI - Effects of short-term cooling on pregnancy rate of dairy heifers under summer heat stress. AB - Thermal stress in cattle results in major decreases in dairy production and reproduction. A study was designed to evaluate the influence of temporary cooling on pregnancy rate in Holstein heifers during summer heat stress. Estrus was synchronized with two injections of prostaglandin [Formula: see text] (PG), administrated 11 d apart, and all heifers were housed in a shaded enclosed structure at the time of the second PG injection. After estrus detection, heifers were randomly divided into three groups: Control (C; n = 30), Sprinkler (S; n = 30) or Sprinkler and Fan (SF; n = 30). Rectal temperatures were measured 2.5 hours before AI, at the time of AI, and 1.5 and 3.5 hours after AI. Group C heifers receive no further treatment, but heifers in S and SF groups were exposed to short-term cooling from 2 hours before until 2 hours after AI with a sprinkler (S) or sprinkler and a fan (SF), respectively. Estrus detection and AI were performed by a single skilled technician; semen was from a bull of known fertility. Rectal temperature did not differ among groups 2.5 hours before AI, but at the time of AI was lower (P < 0.05) in SF group (39.3 +/- 0.0 degrees C) than in S (39.5 +/- 0.0 degrees C) and C (39.9 +/- 0.1 degrees C) groups which also differed (P < 0.05). At 1.5 and 3.5 hours after AI, rectal temperature remained lower in SF group (38.9 +/- 0.0 and 38.7 +/- 0.0 degrees C, respectively) than in the S (39.4 +/- 0.0 and 39.2 +/- 0.0 degrees C, respectively) and C (39.3 +/- 0.0 and 39.3 +/- 0.0 degrees C, respectively) groups, which no longer differed. Pregnancy rate following AI was higher (P < 0.05) in SF group (56.7%) than in the C group (23.3%) with the S group (40%) intermediate and not different from either. The present study results indicate that cooling of dairy heifers for a short time before and after AI, especially with sprinkler and fan, can increase pregnancy rate during heat stress. PMID- 19219563 TI - Trypanosomosis of cattle in selected districts of Awi zone, northwestern Ethiopia. PMID- 19219564 TI - Causal criteria and the problem of complex causation. AB - Nancy Cartwright begins her recent book, Hunting Causes and Using Them, by noting that while a few years ago real causal claims were in dispute, nowadays "causality is back, and with a vengeance." In the case of the social sciences, Keith Morrison writes that "Social science asks 'why?'. Detecting causality or its corollary-prediction-is the jewel in the crown of social science research." With respect to the health sciences, Judea Pearl writes that the "research questions that motivate most studies in the health sciences are causal in nature." However, not all data used by people interested in making causal claims come from experiments that use random assignment to control and treatment groups. Indeed, much research in the social and health science depends on non experimental, observational data. Thus, one of the most important problems in the social and health sciences concerns making warranted causal claims using non experimental, observational data; viz., "Can observational data be used to make etiological inferences leading to warranted causal claims?" This paper examines one method of warranting causal claims that is especially widespread in epidemiology and the health sciences generally-the use of causal criteria. It is argued that cases of complex causation generally, and redundant causation-both causal overdetermination and causal preemption-specifically, undermine the use of such criteria to warrant causal claims. PMID- 19219565 TI - Under the pretence of autonomy: contradictions in the guidelines for human tissue donation. AB - The paper concerns the uncertainty in current propositions for the regulation of tissue donation. It focuses mainly on two statements issued in Germany. The scope of the paper is to give a systematic approach to ethical problems coming up in this field. Both statements try to maintain the idea of positive autonomy in regard to tissue donation, but their attempt eventually is forced to fail. Different procedures are proposed that most often are not practicable (because a truly "informed" consent is impossible, or because optional models tend to overwhelm donors). Blanket consent is also proposed, but this form of consent cannot be seen as an expression of self-determination. Under the pretence of autonomy, donors are left alone with the task to control scientific research and to have their personal and property rights respected. Following this rather weak position of autonomy, in one statement there is a clear tendency to place the intrinsic value of research over the autonomy of the donor. In order to avoid this conclusion, autonomy has to be more than individual decision making. It has to be embedded in social and institutional rules which support and protect individual choice. If the benefits of research are supposed to lead to a common good and to satisfy public interests, then research has to be controlled through public institutions. Autonomy does not exclude institutional support, as institutional support is the only way to take the autonomy of donors seriously. PMID- 19219566 TI - Target selection and annotation for the structural genomics of the amidohydrolase and enolase superfamilies. AB - To study the substrate specificity of enzymes, we use the amidohydrolase and enolase superfamilies as model systems; members of these superfamilies share a common TIM barrel fold and catalyze a wide range of chemical reactions. Here, we describe a collaboration between the Enzyme Specificity Consortium (ENSPEC) and the New York SGX Research Center for Structural Genomics (NYSGXRC) that aims to maximize the structural coverage of the amidohydrolase and enolase superfamilies. Using sequence- and structure-based protein comparisons, we first selected 535 target proteins from a variety of genomes for high-throughput structure determination by X-ray crystallography; 63 of these targets were not previously annotated as superfamily members. To date, 20 unique amidohydrolase and 41 unique enolase structures have been determined, increasing the fraction of sequences in the two superfamilies that can be modeled based on at least 30% sequence identity from 45% to 73%. We present case studies of proteins related to uronate isomerase (an amidohydrolase superfamily member) and mandelate racemase (an enolase superfamily member), to illustrate how this structure-focused approach can be used to generate hypotheses about sequence-structure-function relationships. PMID- 19219567 TI - Rosmarinic acid inhibits 6-OHDA-induced neurotoxicity by anti-oxidation in MES23.5 cells. AB - Rosmarinic acid (RA) is a naturally occurring polyphenolic compound. It is found in several herbs in the Lamiaceae family, such as Perilla frutescens. RA has been reported to exert anti-oxidative effects on rat erythrocyte, liver, and kidney cells. However, little is known about the effects of RA on dopaminergic cells. In the present study, we investigated whether RA could protect MES23.5 dopaminergic cells from 6-hydroxydopamine (6-OHDA)-induced neurotoxicity. The results showed that RA pretreatment significantly prevented 6-OHDA-induced cell viability reduction. Further experiments demonstrated that 6-OHDA induced intracellular reactive oxygen species generation and decreased the mitochondria membrane potential (DeltaPsim). These effects could be partially reversed by RA pretreatment. However, RA had no direct chemical reaction with 6-OHDA extracellularly in a cell-free system. Taken together, these results suggest that RA could exert its protective effects against 6-OHDA-induced neurotoxicity through its anti-oxidation properties. Thus, we propose that RA should be viewed as a potential chemotherapeutic in Parkinson's disease patients. PMID- 19219568 TI - Early prediction of the effectiveness of antidepressants: inputs from an animal model. AB - Although depressive disorders affect approximately 5% of the population in developed countries each year, current antidepressants usually require several weeks to produce beneficial clinical effects and are only effective in about 55% of patients. Therefore, early prediction of the effectiveness of a particular antidepressant for a patient is important for effective pharmacological treatment of depression. In this study, we examined a new method, based on mathematical analyses, of exploratory behavior for predicting the effectiveness of particular antidepressants shortly after initiation of treatment. By using this method, we were able to predict the effectiveness of antidepressants 1-3 days after initiation of treatment in individual subjects. PMID- 19219569 TI - HHH regime for arteritis secondary to TB meningitis: a prospective randomized study. AB - BACKGROUND: Tuberculous meningitis (TBM) is a fairly common, debilitating disease and is often complicated by arteritis resulting in brain infarction. Few treatment regimes specifically address this condition. Hypervolemia-hypertension hemodilution (HHH) regime is known to be effective for treatment of vasospasm complicating subarachnoid hemorrhage. We studied the efficacy of HHH regime in patients with TBM with arteritis using a prospective, randomized study design. PATIENTS AND METHODS: Patients diagnosed to have TB meningitis by clinical, CSF, and imaging findings were evaluated for arteritis, which was recognized by presence of focal neurologic deficits with or without corresponding focal hypodensities on brain CT scan. Patients with deficits of < 96 h were randomized to HHH or conservative treatment. All patents received four-first-line anti-TB drugs and Inj.dexamethasone. HHH therapy was administered over 3-9 days. Neurologic status and modified Rankin score were noted serially and at discharge. RESULTS: Seven patients received HHH and 5, conservative treatment. All had hemiparesis with power 0-3/5. Median GCS was worse in HHH group (11 vs. 13). In the HHH group, 6/7 improved in motor power, and 5/7 in sensorium. In the control group, 3/5 improved in motor power and 3/5 in sensorium. Four patients died in each group. CONCLUSION: HHH therapy is safe and may be beneficial in the management of patients with infective arteritis secondary to TBM. Further study in a larger group with improved monitoring of cerebral circulation is indicated. PMID- 19219570 TI - Estrogen modulates expression of putative housekeeping genes in the mouse uterus. AB - Estrogens regulate gene expression and cell proliferation in target tissues. In studies of estrogen-regulated gene expression, identification of appropriate housekeeping genes (HKGs), reference genes whose expression is not altered by treatment, is difficult. The goal of this study was to define HKGs unaltered by estrogen in the mouse uterus. Ovariectomized C57BL6 mice were dosed with 20 micrograms/kg ethinylestradiol and the uterus was collected at 6, 24, and 72 h later to bracket the biphasic time course of estrogen action in the rodent uterus. RNA was isolated, cDNA synthesized and equal amounts of cDNA were added to real-time PCR reactions. The expression of seven out of nine putative HKGs was altered by estrogen in the mouse uterus. Estrogen induced four gene expression profiles, expression of: (1) Actb and Hsp90ab1 were up-regulated early, (2) B2m and Gusb were up-regulated late, (3) Gapdh, Hprt1, and Ppia were up regulated at all time points, and (4) Rpl13a and 18srRNA were unaltered. This highlights the need to empirically determine the appropriate HKG for each experimental condition. Based on these results, we suggest using Rpl13a or 18srRNA as HKGs for xenoestrogen studies in the mouse uterus and as good candidates to test under different experimental conditions. PMID- 19219573 TI - A new clinical sign in Holmes-Adie syndrome. PMID- 19219572 TI - Morphological differences in Parkinson's disease with and without rest tremor. AB - BACKGROUND: Rest tremor is a hallmark of Parkinson's disease (PD), but its pathogenesis remains incompletely understood. Nigro-striatal dopamine deficiency correlates best with bradykinesia, but not with tremor. Oscillating neurons in one or multiple localizations within the basal gangliathalamo-cortical loop may cause rest tremor, and an active contribution of the cerebellum and the cerebello thalamo-cortical projections has been postulated. OBJECTIVE: To compare the pattern of grey matter volume in PD patients with and without tremor to identify structural correlates of rest tremor. METHODS: Voxel-based morphometry (VBM) of a high-resolution 3 Tesla, T1-weighted MR images, pre-processed according to an optimized protocol using SPM2, was performed in 24 patients with mild to moderate PD comparing local grey matter volume in patients with (n = 14) and without rest tremor (n = 10). RESULTS: Grey matter volume is decreased in the right quadrangular lobe and declive of the cerebellum in PD with tremor compared to those without (PFDR < 0.05). CONCLUSIONS: These results demonstrate for the first time morphological changes in the cerebellum in PD patients with rest tremor and highlight the involvement of the cerebellum and cerebello- thalamo-cortical circuit in the pathogenesis of parkinsonian rest tremor. PMID- 19219574 TI - Steroid-unresponsive neurosarcoidosis successfully treated with adalimumab. PMID- 19219575 TI - Severe and rapidly evolving peripheral neuropathy revealing sporadic Creutzfeldt Jakob disease. PMID- 19219576 TI - Iranian beet necrotic yellow vein virus (BNYVV): pronounced diversity of the p25 coding region in A-type BNYVV and identification of P-type BNYVV lacking a fifth RNA species. AB - Beet necrotic yellow vein virus (BNYVV) was detected in 288 of the 392 samples collected in Iran. A-type BNYVV was detected most frequently. The p25 coding region on BNYVV RNA-3 was amplified by RT-PCR and sequenced. Nine different variants of the highly variable amino acid tetrad at positions 67-70 of p25 were identified, i.e. ACHG, AHHG, AYHG, ALHG, AFHR, AFHG, AHYG, VLHG and VHHG. These are more different tetrad variants than have been reported from any other country. The first three variants were found most commonly. In 23 out of the 288 BNYVV-positive samples, we detected P-type BNYVV that had previously been identified only in France, Kazakhstan and recently in the UK. Surprisingly, none of these samples contained the fifth RNA species usually associated with P-type BNYVV in other countries. As in other BNYVV P-type sources, the p25 amino acid tetrad in positions 67-70 of the Iranian P-type consists of SYHG. PMID- 19219577 TI - Synthesis of orthogonally protected azahistidine: application to the synthesis of a GHK analogue. AB - The synthesis of various orthogonally protected azahistidine derivatives are obtained via 1,3-dipolar cycloaddition reactions. The newly obtained amino-acids can be selectively deprotected either at the side chain or at the N-terminus of the amino acid and should thus allow the use of these derivatives in (solid phase) peptide synthesis. PMID- 19219578 TI - Phylogeny of Litsea and related genera (Laureae-Lauraceae) based on analysis of rpb2 gene sequences. AB - The relationship between Litsea and related genera is currently unclear. Previous molecular studies on these taxa using cpDNA and nrITS were unable to produce well resolved phylogenetic trees. In this study, we explored the potential of the rpb2 gene as a source of molecular information to better resolve the phylogenetic analysis. Although rpb2 was believed to be a single-copy gene, our cloning results showed that most species examined possessed several copies of these sequences. However, the genetic distance among copies from any one species was low, and these copies always formed monophyletic groups in our molecular trees. Our phylogenetic analyses of rpb2 data resulted in better resolved tree topologies compared to those based on cpDNA or nrITS data. Our results show that monophyly of the genus Litsea is supported only for section Litsea. As a genus, Litsea was shown to be polyphyletic. The genera Actinodaphne and Neolitsea were resolved as monophyletic groups in all analyses. They were also shown to be sisters and closer to the genus Lindera than to the genus Litsea. Our results also revealed that the genus Lindera is not a monophyletic group. PMID- 19219580 TI - Multifarious proteomic signatures and regional heterogeneity in glioblastomas. AB - To investigate the underlying intratumoral diversity of molecular profiles in glioblastomas, a proteomic approach was introduced to compare samples from regions of different histological grade. Using two-dimensional gel electrophoresis (2DE) with matrix-assisted laser-desorption/ionization time-of flight mass spectrometry (MALDI-TOF MS), we compared prospectively collected tissue samples of different histological grade areas of three glioblastoma patients. Consistent results showing relatively high expression of ubiquitin carboxyl-terminal esterase L1 in low-histological-grade areas (Grade 2 > Grades 3 and 4) and high expression of transthyretin in high-histological-grade areas (Grade 2 < Grades 3 and 4) were demonstrated. These results were confirmed by western blot (WB) analysis and immunohistochemical staining. This study provided the evidence of multifarious proteomic signatures according to regional and histological heterogeneity in glioblastomas. PMID- 19219581 TI - Functional characterization of kanB by complementing in engineered Streptomyces fradiae Deltaneo6::tsr. AB - A putative aminotransferase gene, kanB, lies in the biosynthetic gene cluster of Streptomyces kanamyceticus ATCC 12853 and has 66% identity with neo6 in neomycin biosynthesis. Streptomyces fradiae Deltaneo6::tsr was generated by disrupting neo6 in the neomycin producer Streptomyces fradiae. Neomycin production was completely abolished in the disruptant mutant but was restored through self complementation of neo6. S. fradiae HN4 was generated through complementation with kanB in Streptomyces fradiae neo6::tsr. Based on metabolite analysis by ESI/MS and LC/MS, neomycin production was restored in Streptomyces fradiae HN4. Thus, like neo6, kanB also functions as a 2-deoxy-scyllo-inosose aminotransferase that has dual functions in the formation of 2-deoxy-scyllo-inosose (DOS). PMID- 19219583 TI - Overview of ELISA in relation to other disciplines. PMID- 19219584 TI - Systems in ELISA. AB - This chapter defines the terms and examines the configurations used for most applications of ELISA. Such a chapter is important because the possibilities inherent in the systems of ELISA must be understood in order to maximize their versatility in assay design. All heterogeneous systems have three basic parameters: 1. One reactant is attached to a solid phase, usually a plastic microtiter plate with an 8 x 12-well format. 2. Separation of bound and free reagents, which are added subsequently to the solid phase-attached substance, is by a simple washing step. 3. Results are obtained through the development of color. PMID- 19219586 TI - Titration of reagents. AB - This chapter examines in detail the necessary practical skills needed to facilitate the development and sustainability of ELISAs. Basic methodologies to set up all systems are discussed so that the reader can both investigate the possibilities of tests using his or her reagents and use with confidence the reagents obtained from other sources. As with all other tests, it is vital that scientists realize the principles of the methods to allow good judgement based on quantifiable and controllable features. In other words, there should be a full understanding of what is being performed in the laboratory. The intention of this training is to produce new skills and understanding as well as to encourage critical assessment. Such a mentality is necessary and supported through the application of statistical criteria to results and to continuous monitoring of performance. PMID- 19219585 TI - Stages in ELISA. AB - This chapter gives general information on essential practical features of ELISAs. These can be summarized as follows: 1. Adsorption of antigen or antibody to the plastic solid phase. 2. Addition of the test sample and subsequent reagents. 3. Incubation of reactants. 4. Separation of bound and free reactants by washing. 5. Addition of enzyme-labeled reagent. 6. Addition of enzyme detection system (color development). 7. Visual or spectrophotometric reading of the assay. PMID- 19219587 TI - Theoretical considerations. AB - This chapter examines the aspects of using ELISA to solve problems, definitions of terms met in serology, antibody structure, and the production of antibodies in animals, units, dilutions, and molarities. Antibodies--A Laboratory Manual (1) is an excellent manual of techniques relevant to ELISA, and all scientists involved in experimental work involving antibodies should have this manual. The manuals given in refs. 2 and 3 also provide extensive relevant practical information. PMID- 19219588 TI - Practical exercises. AB - The aim of this chapter is to illustrate the principles of ELISA by (1) showing worked examples of each assay, including diagrams of plates and representational data from assays, (2) analyzing such data in terms of important rules that are learned at each stage, and (3) providing full working instructions for investigators to perform each assay so that they obtain their own data to be analyzed. This chapter can be used in several ways. First, researchers without access to reagents will obtain a working knowledge of ELISA through the examples. Second, it can be used in training courses in which reagents may be provided (as indicated in the text). Third, the information will be useful for investigators who have already had some experience with the technique but may have had difficulties in obtaining and analyzing data. Remember that it is the application of ELISA to specific problems, and not the methodology for its own sake, that is the most important reason the techniques should be mastered. PMID- 19219589 TI - Monoclonal antibodies. AB - The ability to produce and exploit monoclonal antibodies (mAbs) has revolutionized many areas of biological sciences. The unique property of an mAb is that it is a single species of immunoglobulin (IG) molecule. This means that the specificity of the interaction of the paratopes on the IG, with the epitopes on an antigenic target, is the same on every molecule. This property can be used to great benefit in immunoassays to provide tests of defined specificity and sensitivity, which improve the possibilities of standardization. The performance of assays can often be determined relating the actual weight of antibody (hence the number of molecules) to the activity. Often the production of an mAb against a specific epitope is the only way that biological entities can be differentiated. This chapter outlines the areas involving the development of assays based on mAbs. The problems involved address include the physical aspects of mAbs and how they may affect assay design and also the implications of results based on monospecific reagents. Often these are not fully understood, leading to assays that are less than satisfactory, which does not justify the relatively high cost of preparing and screening of mAbs. There are many textbooks and reviews dealing with the preparation of mAbs, the principles involved, and various purification and manipulative methods for the preparation of fragments and conjugation. There has been little general information attempting to summarize the best approaches to assay design using mAbs. Much time can be wasted through bad planning, and this is particularly relevant to mAbs. A proper understanding of some basic principles is essential. It is beyond the scope of this chapter to discuss all aspects, but major areas are highlighted. PMID- 19219590 TI - Validation of diagnostic tests for infectious diseases. PMID- 19219591 TI - Charting methods for internal quality control for competition ELISA. AB - This chapter deals with relatively simple ways to use control charts to monitor the performance of ELISAs. A rinderpest competition ELISA, for the estimation of antibodies in serum samples, is used to demonstrate the methods. This assay is available in a kit form. Constant evaluation of the use of the kit is part of what is called internal quality control (IQC). Figure 1 shows an overview of the ELISA scheme described in this chapter. The details of the procedure, which involves plotting the data graphically (charting methods), are explained herein. The objectives of charting data are as follows: 1. To keep a constant record of all data. 2. To monitor the assay from plate to plate in any one day's testing. 3. To monitor the tests made from day to day, week to week, year to year. 4. To allow rapid identification of unacceptable results. 5. To allow recognition of reagent problems. 6. To identify trends in results (increasingly poor performance). 7. To identify when a new set of kit reagents is necessary. 8. To allow identification of differences among operators of the assay. 9. To fulfill various criteria for good laboratory practice. 10. To fulfill necessary requirements for external recognition to prove that tests are being performed at an acceptable level (increasingly important when results are used for international trading purposes). PMID- 19219592 TI - Charting methods for internal quality control of indirect ELISA. AB - This chapter deals with control charts to monitor the performance of Indirect ELISAs. An Indirect ELISA kit for the detection of antibodies against Brucella is used to demonstrate the methods. Many of the features explained in Chapter 9 are relevant to this chapter; some repetition is intended, as this chapter may be read independently. Figure 1 gives an overview of the indirect ELISA scheme used. The details of the procedure, which involves plotting the data graphically (charting methods), are explained. As a reminder, the objectives of charting data are as follows: 1. To keep a constant record of all data. 2. To monitor the assay from plate to plate in any one day's testing. 3. To monitor the tests made from day to day, week to week, year to year. 4. To allow rapid identification of unacceptable results. 5. To allow recognition of reagent problems. 6. To identify trends in results (increasingly poor performance). 7. To identify when a new set of kit reagents is necessary. 8. To allow identification of differences in operators of the assay. 9. To fulfill various criteria for good laboratory practice. 10. To fulfill necessary requirements for external recognition that tests are being performed at an acceptable level (increasingly important when results are used for international trading purposes). PMID- 19219593 TI - Ruggedness and robustness of tests: aspects of kit use and validation. PMID- 19219594 TI - More advanced statistical methods for quality assurance, test validation, and interpretation. PMID- 19219595 TI - Internal quality control and external quality management of data in practice. AB - In this chapter, the use of control charts to both continuously evaluate testing in individual laboratories as well as provide data for external monitoring is examined in detail. The data is based on the publication by D. E. Rebeski' et al., "Charting methods to monitor the operational performance of ELISA method for the detection of antibodies against trypanosomes" in Veterinary Parasitology, 2001, 96, 11-50, and is a detailed example of the investigation of the performance of four indirect ELISAs for the detection of antibodies against trypanosomes using Trypanosoma congolense and T. vivax antigen-precoated plates in 15 veterinary diagnostic laboratories in Africa and Europe. The study shows the practical use of charting methods with respect to assessing the operational performance of each ELISA. Data from standardized internal quality control (IQC) samples were used to assess ELISA performance indicators with reference to expected upper and lower control limits, as determined from studies by the kit producer (tentative values). Based on unprocessed (optical density) and normalized absorbance values (calculated as a percentage positivity of a control), dispersion of values from the expected data range were estimated though plotting the location and deviation of the values. In addition, assay precision was estimated by plotting the distribution of coefficients of variation <10% of the IQCs. Binding ratios of various controls were calculated to estimate the assay proficiency with respect to the accuracy of assessing whether the IQC samples tested positive or negative in the test proper. The graphical analysis of dispersion of absorbance values in combination with assay precision and proficiency criteria were considered satisfactory to allow the evaluation of the operational performances of the ELISAs, and provide useful decision-making criteria for plate acceptance and rejection. The establishment of standardized and transparent IQC data charting methods for the indirect ELISAs provided an increased measure of confidence to national laboratories with respect to their reports on disease occurrence. Moreover, the relative assay performances among all laboratories were examined, using summary data charts, with reference to the performance criteria described. The IQC data were also examined using modified Youden plot analysis, demonstrating that indirect ELISA methods can be successfully applied at diagnostic laboratories in the tropics for monitoring trypanosomosis control programs. PMID- 19219596 TI - Immunochemical techniques. AB - The scope of this book does not allow a complete description of the many techniques available for purification and treatment of reagents for facilitating immunoassays in general. There is a large amount of literature covering techniques, and this can be consulted for specific problems. The examination of many of the catalogs produced by commercial companies is useful as they often include good technical sections describing methods using their products. This chapter contains the practical basics of conjugation (a large field in itself), and details other immediately useful techniques that might be first desired in starting an ELISA. The book Antibodies: A Laboratory Manual (1) should be regarded as definitive in the laboratory because it is extremely "digestible" and covers a large field of methods, all of which are relevant to ELISA. PMID- 19219598 TI - ps-2, the gene responsible for functional sterility in tomato, due to non dehiscent anthers, is the result of a mutation in a novel polygalacturonase gene. AB - The recessive mutation ps-2, which appeared spontaneously in tomato, confers functional male sterility due to non-dehiscent anthers. In this study, we isolated and characterized the PS-2 gene. A single nucleotide mutation in a novel tomato polygalacturonase gene is responsible for the ps-2 phenotype. The mutation in ps-2 is responsible for an alternative splicing during maturation of the pre mRNA, which leads to an aberrant mRNA. Differentiation between ps-2 and wild type (PS-2) anthers only appears in the final developmental stage in which the stomium remains closed in the mutant. To our knowledge, this is the first functional sterility gene isolated in the Solanaceae family. The specific expression of the Arabidopsis homolog of PS-2 in the anther dehiscence zone suggests a conserved mode of action over the plant kingdom, which means that the repression of PS-2 homologs may be a potential way to introduce functional sterility in other species. PMID- 19219599 TI - QTL analysis of plant development and fruit traits in pepper and performance of selective phenotyping. AB - A QTL analysis was performed to determine the genetic basis of 13 horticultural traits conditioning yield in pepper (Capsicum annuum). The mapping population was a large population of 297 recombinant inbred lines (RIL) originating from a cross between the large-fruited bell pepper cultivar 'Yolo Wonder' and the small fruited chilli pepper 'Criollo de Morelos 334'. A total of 76 QTLs were detected for 13 fruit and plant traits, grouped in 28 chromosome regions. These QTLs explained together between 7% (internode growth time) and 91% (fruit diameter) of the phenotypic variation. The QTL analysis was also performed on two subsets of 141 and 93 RILs sampled using the MapPop software. The smaller populations allowed for the detection of a reduced set of QTLs and reduced the overall percentage of trait variation explained by QTLs. The frequency of false positives as well as the individual effect of QTLs increased in reduced population sets as a result of reduced sampling. The results from the QTL analysis permitted an overall glance over the genetic architecture of traits considered by breeders for selection. Colinearities between clusters of QTLs controlling fruit traits and/or plant development in distinct pepper species and in related solanaceous crop species (tomato and eggplant) suggests that shared mechanisms control the shape and growth of different organs throughout these species. PMID- 19219600 TI - Treatment of symptomatic complex posterior circulation cerebral artery stenosis with balloon-mounted stents: technique feasibility and outcome. AB - PURPOSE: This study aimed to retrospectively analyze a series of patients with complex posterior circulation stenosis who underwent stent-assisted angioplasty to evaluate the feasibility of the procedure and summarize the experience with regard to complications. METHODS: A total of 16 consecutive patients with 27 complex posterior circulation artery stenoses refractory to medical therapy were enrolled. Technical success rate, periprocedural complication, and long-term follow-up result were evaluated. RESULTS: The study population included 16 patients with 27 lesions. A total of 36 stents were successfully implanted. The technical success rate was 100%, and the overall periprocedural complication rate was 12.5% (2/16). During a median of 25.5 months of follow-up, three patients presented recurrent transient ischemic attacks, which were confirmed had restenosis more than 50% by control angiography or transcranial Doppler. CONCLUSIONS: Stent-assisted angioplasty is a feasible treatment method for complex posterior circulation artery stenosis. However, it appears to be associated with a relatively high periprocedural complication rate. Therefore, strict periprocedural management to reduce mortality and morbidity is needed. PMID- 19219601 TI - Brain edema associated with unruptured brain arteriovenous malformations. AB - INTRODUCTION: Brain edema in unruptured brain arteriovenous malformations (AVMs) is rare; this study examines (1) its frequency and clinical presentation, (2) imaging findings with emphasis on venous drainage abnormalities, and (3) implications of these findings on natural history and management. METHODS: Presentation and imaging features of all unruptured brain AVMs were prospectively collected in our brain AVM database. Neurological findings, size, location, venous drainage pattern, presence of venous thrombosis, ectasia, or stenosis, and brain edema were specifically recorded. Treatment details of all patients with brain edema and their clinical and imaging follow-up were reviewed. Finally, a comparison was made between patients with and without edema. RESULTS: Brain edema was found in 13/329 unruptured brain AVMs (3.9%). Neurological deficit (46.2%), venous thrombosis (38.5%), venous ectasia (84.6%), stenosis (38.5%), and contrast stagnation in the draining veins (84.6%) were more frequent in patients with brain edema than without edema. Eight patients with brain edema received specific treatment (embolization = 5, surgery = 2, radiosurgery = 1). Clinical features correlated well with change in degree of edema in six. Three of five embolized patients were stable or showed improvement after the procedure. On follow-up, however, intracranial hemorrhage developed in three. CONCLUSION: Brain edema in unruptured brain AVMs is rare, 3.9% in this series. Venous outflow abnormalities are frequently associated and appear to contribute to the development of edema. Progressive nonhemorrhagic symptoms are also associated, with a possible increased risk of hemorrhage. Palliative embolization arrests the nonhemorrhagic symptoms in selected patients, although it may not have an effect on hemorrhagic risk. PMID- 19219602 TI - Prostaglandin synthase 2/cyclooxygenase 2 (PTGS2/COX2) 8473T>C polymorphism associated with prognosis for patients with colorectal cancer treated with capecitabine and oxaliplatin. AB - PURPOSE: The present study analyzed the polymorphisms of apoptosis-related genes and their impact on the response to chemotherapy and survival of patients with colorectal cancer. PATIENTS AND METHODS: A total of 76 patients with recurrent or metastatic colorectal cancer treated with capecitabine and oxaliplatin (XELOX) combination chemotherapy were enrolled in the present study. The single nucleotide polymorphisms of 15 apoptosis-related genes (TP53, BCL2L, TNFRSF10B, AKT1, PTGS2/COX2, BID, RIPK1, FAS, FASL, caspase 3, and caspase 6-10) were determined using a PCR-RFLP assay. RESULTS: No significant association between the polymorphisms and the response was found for any of the genes analyzed. However, the T/T genotype of PTGS2 8473T>C (rs5275) was significantly correlated with a better progression-free survival (PFS) and overall survival (OS) when compared to the combined T/C and C/C genotype (Hazard ratio [HR] = 0.47; P value = 0.046 and HR = 0.16; P = 0.013, respectively) in a multivariate analysis adjusted for age, sex, performance status, disease status and curative resection. No association was noted between the other polymorphisms and survival. CONCLUSION: The PTGS2 8473T>C polymorphism was found to be correlated with PFS and OS in patients with advanced colorectal cancer treated with XELOX chemotherapy. PMID- 19219603 TI - Analysis of feasibility and toxicity of concurrent chemoradiotherapy with S-1 for locally advanced squamous cell carcinoma of the head and neck in elderly cases and/or cases with comorbidity. AB - PURPOSE: The aim of this study was to evaluate the feasibility and toxicity of concurrent chemoradiotherapy (CCRT) with S-1 in patients with locally advanced squamous cell carcinoma of the head and neck (SCCHN) in elderly cases and/or cases with comorbidity. METHODS: Fifty eligible patients with stage III (15 cases) or stage IV (35 cases) SCCHN were treated with CCRT. Thirteen cases had an advanced age of over 75 years and 37 cases had comorbidity. Definitive radiotherapy was delivered up to a total dose of 66-70.2 Gy. The patients received two courses of oral S-1 (40 or 50 mg twice a day [80 or 100 mg/day]) for 2 weeks followed by 1 week of rest while receiving CCRT. RESULTS: All the patients received the planned radiotherapy and at least one course of S-1. Grade 3 mucositis occurred in 20% of the patients (10/50). Grade 3 neutropenia occurred in 12% (6/50) and leukocytopenia occurred in 6% (3/50) of the cases. Pathologically, the complete response rates were 93% in stage III and 54% in stage IV. CONCLUSION: Concurrent chemoradiotherapy with S-1 is a safe, well tolerated and effective regimen for locally advanced SCCHN in elderly cases and/or cases with comorbidity. PMID- 19219604 TI - Long-term results of gemcitabine plus oxaliplatin with and without rituximab as salvage treatment for transplant-ineligible patients with refractory/relapsing B cell lymphoma. AB - PURPOSE: To determine the efficacy and safety of the combination of gemcitabine plus oxaliplatin, with and without rituximab, in patients with relapsed/refractory B-cell lymphoma unsuitable for high dose therapy. METHODS: Patients were prospectively enrolled in two subsequent trials, GEMOX [gemcitabine (1200 mg/m(2), days 1 and 8) and oxaliplatin (120 mg/m(2), day 2), three-weekly] and R-GEMOX [rituximab (375 mg/m(2), day 1), gemcitabine (1200 mg/m(2), day 1) and oxaliplatin (120 mg/m(2), day 2), bi-weekly], up to six courses. RESULTS: Sixty-two patients were enrolled: GEMOX [n = 30; median age, 66 years (range, 46 85); previous chemotherapy > or =2, 70%; PS ECOG > or = 2, 57%]; R-GEMOX [n = 32; median age, 65 years (range 32-79); previous chemotherapy > or =2, 75%; PS ECOG > or = 2, 47%]. Overall and complete response rates were 57 and 30% (95% CI, 15-49) for GEMOX and 78 and 50% (95% CI, 32-68) in R-GEMOX, respectively. Grade 3/4 neutropenia occurred in 57 and 47% of cycles and grade 3/4 thrombocytopenia in 26 and 17% of courses for GEMOX and R-GEMOX, respectively. At 42 months, the failure free survival (FFS) was 7% (95% CI, 0-16) for GEMOX and 28% (95% CI, 9-47) for R GEMOX (P = 0.014), with overall survivals of 7 (95% CI, 0-16) and 37% (95% CI, 20 55), respectively (P = 0.016). CONCLUSIONS: Both regimes showed good tolerability and appealing response rates. FFS was more prolonged in R-GEMOX, but patients continuously relapsed without a clear plateau on survival curves. PMID- 19219606 TI - Information-gathering patterns associated with higher rates of diagnostic error. AB - Diagnostic errors are an important source of medical errors. Problematic information-gathering is a common cause of diagnostic errors among physicians and medical students. The objectives of this study were to (1) determine if medical students' information-gathering patterns formed clusters of similar strategies, and if so (2) to calculate the percentage of incorrect diagnoses in each cluster. A total of 141 2nd year medical students completed a computer case simulation. Each student's information-gathering pattern included the sequence of history, physical examination, and ancillary testing items chosen from a predefined list. We analyzed the patterns using an artificial neural network and compared percentages of incorrect diagnoses among clusters of information-gathering patterns. We input patterns into a 35 x 35 self organizing map. The network trained for 10,000 epochs. The number of students at each neuron formed a surface that was statistically smoothed into clusters. Each student was assigned to one cluster, the cluster that contributed the largest value to the smoothed function at the student's location in the grid. Seven clusters were identified. Percentage of incorrect diagnoses differed significantly among clusters (Range 0-42%, Chi (2) = 13.62, P = .034). Distance of each cluster from the worst performing cluster was used to rank clusters. This rank was compared to rank determined by percentage incorrect. We found a high positive correlation (Spearman Correlation = .893, P = .007). Clusters closest to the worst performing cluster had the highest percentages of incorrect diagnoses. Patterns of information-gathering were distinct and had different rates of diagnostic error. PMID- 19219605 TI - A mathematical model of blood, cerebrospinal fluid and brain dynamics. AB - Using first principles of fluid and solid mechanics a comprehensive model of human intracranial dynamics is proposed. Blood, cerebrospinal fluid (CSF) and brain parenchyma as well as the spinal canal are included. The compartmental model predicts intracranial pressure gradients, blood and CSF flows and displacements in normal and pathological conditions like communicating hydrocephalus. The system of differential equations of first principles conservation balances is discretized and solved numerically. Fluid-solid interactions of the brain parenchyma with cerebral blood and CSF are calculated. The model provides the transitions from normal dynamics to the diseased state during the onset of communicating hydrocephalus. Predicted results were compared with physiological data from Cine phase-contrast magnetic resonance imaging to verify the dynamic model. Bolus injections into the CSF are simulated in the model and found to agree with clinical measurements. PMID- 19219607 TI - Serum concentrations of formation (PINP) and resorption (Ctx) bone turnover markers in rheumatoid arthritis. AB - Joint inflammation in rheumatoid arthritis (RA) induces local periarticular osteoporosis. Generalised bone mineral density (BMD) decrease concerns approximately 50% of rheumatic patients. Both types of bone mass depletion can issue from cytokine-induced (TNF-alpha, IL-1, IL-6) osteoclasts' activation, osteoprotegerin and its ligand's (RANKL) function disorders, patients' immobilisation and glucocorticosteroid (GCS) intake, as well as from hormonal alterations in postmenopausal women, predominate among RA individuals. The aim of the study was to compare serum concentrations of marker of bone formation--serum aminoterminal propeptide of type I collagen (PINP), and bone resorption, carboxy (C) terminal telopeptide (Ctx), bone turnover markers in RA and osteoarthritis (OA) patients and in RA groups of different disease activity, different degree of joint damage and the history of GCS intake. A total of 50 RA female patients and 50 women with knee OA were included in the study. Blood for morphology and biochemistry laboratory tests was taken. Joint X-rays to establish OA and RA diagnosis and the degree of RA progression, as well as DEXA BMD measurements were performed. PINP and Ctx concentrations were assessed. In RA patients the number of swollen and painful joints, the duration of morning stiffness, visual analogue scale values and Waaler-Rose's test activity were recorded. The Disease Activity Index (DAS 28) was counted from the appropriate formula. No differences in bone turnover markers' concentrations were noted neither between RA and OA patients nor between the RA group when compared to the one without the history of GCS use. Bone turnover markers' concentrations in RA were proportional to the number of swollen and painful joints. However, no correlation was found between the markers' concentrations and RA activity assessed by DAS 28 or by laboratory means. Ctx concentrations were higher in patients at II degree joint damage according to Larsen and Dale's than at more advanced stages. Ctx concentrations decreased with the disease duration. Serum morphogenesis and resorption markers' concentrations change in course of RA indicating the decrease in bone metabolic activity with the disease duration and progression. High RA activity and severity correlate with increased markers' levels-the resorption one. The influence of GCS on bone metabolism in RA requires further study. PMID- 19219608 TI - The application of the mutated acetolactate synthase gene from rice as the selectable marker gene in the production of transgenic soybeans. AB - We investigated selective culturing conditions for the production of transgenic soybeans. In this culturing system, we used the acetolactate synthase (ALS) inhibiting herbicide-resistance gene derived from rice (Os-mALS gene) as a selectable marker gene instead of that derived from bacteria, which interfered with the cultivation and practical usage of transgenic crops. T(1) soybeans grown from one regenerated plant after selection of the ALS-targeting pyrimidinyl carboxy (PC) herbicide bispyribac-sodium (BS) exhibited herbicide resistance, and the introduction and expression of the Os-mALS gene were confirmed by genetic analysis. The selective culturing system promoted by BS herbicide, in which the Os-mALS gene was used as a selectable marker, was proved to be applicable to the production of transgenic soybeans, despite the appearance of escaped soybean plants that did not contain the Os-mALS transgene. PMID- 19219609 TI - Twinned microspore-derived embryos of canola (Brassica napus L.) are genetically identical. AB - Microspore culture is used extensively in several crop species to generate diverse populations of homozygous, doubled haploid lines for breeding and genetic analyses. In our canola (Brassica napus L.) doubled haploid breeding programme we regularly observe conjoined microspore-derived embryos, most commonly twins, joined either at the base of the hypocotyl or along the length of the hypocotyl axis. The aim of this study was to determine if twinned embryos were genetically identical or non-identical in order to gauge their value for breeding and linkage analysis. Microsatellite marker fingerprinting of 12 pairs of twinned embryos produced by microspore culture of heterozygous F(1) lines revealed that pairs of twins were genetically identical. Based on this finding, we recommend breeders and geneticists using microspore culture technology to retain only one embryo from each pair of twinned embryos. PMID- 19219610 TI - The present and future burden of urinary bladder cancer in the world. AB - Urinary bladder cancer (UBC) is a common disease worldwide. At any point in time 2.7 million people have a history of UBC. The incidence of UBC varies over the world with highest rates in developed communities. But the burden of UBC will increase in less developed areas of the world. These changes can be attributed to global changes in exposure to risk factors for UBC and growth and aging of the world population. PMID- 19219611 TI - Experimental rat bladder urothelial cell carcinoma models. AB - Bladder cancer is a major public health problem. Currently available therapeutic options seem to be unable to prevent bladder cancer recurrence and progression. To enable preclinical testing of new intravesical therapeutic agents, a suitable bladder tumor model that resembles human disease is highly desirable. The aim of this topic paper was to discuss the problems associated with current in vivo animal bladder tumor models, focusing on the orthotopic syngeneic rat bladder tumor model. In the second part of the paper the development of a potential new orthotopic rat bladder tumor model is described. PMID- 19219612 TI - Impact of marital status in patients undergoing radical cystectomy for bladder cancer. AB - PURPOSE: Married (vs. unmarried) individuals have improved health status and longer life expectancies in a variety of benign and malignant disease states, including prostate, breast, head/neck, and lung cancers. We sought to evaluate a cohort of patients undergoing cystectomy for bladder cancer to evaluate the impact of marital status on demographic, peri-operative, and pathological outcomes in order to better understand the factors which may contribute to the survival differences observed. METHODS: Two-hundred and two patients underwent radical cystectomy and urinary diversion for bladder cancer. Patients were categorized based on marital status as either married or unmarried (widowed, divorced, never married). Correlations were made to demographic factors (age, race, gender, BMI, tobacco use, alcohol use), perioperative factors (pre-op renal function (creatinine), hematocrit, EBL, hospital stay, choice of diversion), and pathological outcomes (organ-confined status, LN positivity). RESULTS: Of the 202 patients, 74% were married. Married individuals (vs. unmarried) were more often male (84 vs. 62%) and had a higher BMI (28.1 vs. 25.9). Married persons had a significantly lower pre-op creatinine (1.1 vs. 1.4) and higher hematocrit (39 vs. 34). Hospital stay was shorter in married patients by a mean of 1.6 days. Regarding operative pathology, married patients had a higher rate of organ confined disease (59 vs. 47%) (P = 0.05, 0.08 on multivariate) and trended towards a lower rate of LN positivity (15 vs. 21%; P = 0.10, 0.12 multivariate). CONCLUSIONS: In patients undergoing cystectomy for bladder cancer, married individuals appear to have improved pre-operative laboratory variables, shorter hospitalization, and improved pathological outcomes versus unmarried patients in our case series. These findings may support the evidence (observed in other tumor types and other disease states) that married persons present earlier than unmarried individuals, and this may help explain the improved survival outcomes that have been observed in married patients with bladder cancer. PMID- 19219613 TI - Expression of cortactin and survivin in renal cell carcinoma associated with tumor aggressiveness. AB - PURPOSE: We tested the hypothesis that the expression of cortactin and survivin in renal cell carcinomas (RCCs) correlates with more advanced stages of the disease. METHODS: Immunohistochemical analysis of cortactin and survivin expression (scored on a scale of 0-400) was performed in 124 renal cell carcinomas including clear cell renal cell carcinoma (CRCC), papillary RCC (PRCC), CRCC with sarcomatoid differentiation (SRCC), chromophobe RCC (ChRCC), and CRCC with granular cell differentiation (GRCC). RESULTS: Higher cortactin scores in CRCC were significantly correlated with higher T (p = 0.021) and N stages (p = 0.036), and nuclear grade (p = 0.012). Higher cortactin immunostaining scores were associated with higher mortality (p = 0.035). In addition, the survivin scores were significantly higher in the more aggressive GRCC and SRCC than in CRCC, suggesting a significant role of survivin expression in transformation of tumor cells to a more malignant phenotype. CONCLUSIONS: Higher expression of cortactin and survivin significantly correlated with advanced clinicopathological stage. Our findings support the potential targeting of survivin and cortactin for the development of novel therapeutic strategies for renal cell carcinoma. PMID- 19219615 TI - Thermal acclimation to 4 or 10 degrees C imparts minimal benefit on swimming performance in Atlantic cod (Gadus morhua L.). AB - Thermal acclimation is frequently cited as a means by which ectothermic animals improve their Darwinian fitness, i.e. the beneficial acclimation hypothesis. As the critical swimming speed (U (crit)) test is often used as a proxy measure of fitness, we acclimated Atlantic cod (Gadus morhua) to 4 and 10 degrees C and then assessed their U (crit) swimming performance at their respective acclimation temperatures and during acute temperature reversal. Because phenotypic differences exist between different populations of cod, we undertook these experiments in two different populations, North Sea cod and North East Arctic cod. Acclimation to 4 or 10 degrees C had a minimal effect on swimming performance or U (crit), however test temperature did, with all groups having a 10-17% higher U (crit) at 10 degrees C. The swimming efficiency was significantly lower in all groups at 4 degrees C arguably due to the compression of the muscle fibre recruitment order. This also led to a reduction in the duration of "kick and glide" swimming at 4 degrees C. No significant differences were seen between the two populations in any of the measured parameters, due possibly to the extended acclimation period. Our data indicate that acclimation imparts little benefit on U (crit) swimming test in Atlantic cod. Further efforts need to identify the functional consequences of the long-term thermal acclimation process. PMID- 19219614 TI - Extreme tolerance to ammonia fumes in African naked mole-rats: animals that naturally lack neuropeptides from trigeminal chemosensory nerve fibers. AB - Naked mole-rats (Heterocephalus glaber) naturally lack neuropeptides associated with the signaling of chemical irritants from C type trigeminal nerve fibers. The goal of the present study was to assess behavioral responses of these animals to stimulation of the trigeminal chemosensory system, and to determine if stimulation would increase post-synaptic activity in the trigeminal nucleus, as seen in laboratory mice and rats. The results show that naked mole-rats are behaviorally insensitive to capsaicin solution applied to the nostrils and to ammonia fumes in a behavioral avoidance test. Centrally, the number of c Fos labeled cells in the spinal trigeminal nucleus increased from exposure to ammonia although the magnitude of the increase was less than for rats. The increase observed in naked mole-rats likely reflects activity from glutamate release, which appears insufficient to drive pain and aversion behaviors. The results support the idea that neuropeptides in the C fibers of the trigeminal system may be required to signal the aversive quality of specific chemical irritants. The natural lack of neuropeptides in naked mole-rats may be an adaptation to living in a challenging subterranean environment with extremely high levels of ammonia and carbon dioxide, stimuli known to excite trigeminal chemosensory C fibers. PMID- 19219616 TI - A case report of spontaneous rupture of a renal angiomyolipoma in a post-partum 21-year-old patient. AB - INTRODUCTION: Renal angiomyolipomas (AML) are benign tumours containing vascular, smooth muscle and fatty elements. The majority of renal AML run an asymptomatic, benign course. The main associated complication is that of retro-peritoneal or intra-tumoural haemorrhage. Treatment options include conservative management versus interventional procedures such as total or partial nephrectomy, cryotherapy or embolization. CASE: We describe a case of symptomatic, spontaneous rupture of AML in the immediate post-partum period of a patient treated under our care. DISCUSSION: This case highlights the presentation in the form of an acute abdomen in the immediate post-partum period. This is important as acute abdomen following delivery can be attributed to a number of other causes. It also demonstrates that further complications of renal angiomyolipoma rupture can arise, emphasising the importance of post treatment vigilance for signs of infection, further haemorrhage and post embolic events. PMID- 19219617 TI - Primary peritoneal clear cell adenocarcinoma arising in previous abdominal scar for endometriosis surgery. AB - BACKGROUND: Endometriosis-associated ovarian cancer arising from the surgical incision site is an unusual clinical entity. CASE: A 37-year-old woman presented with a chief complaint of progressive swelling of the mons pubis. The patient was status post laparotomy for endometrioma/endometriosis 10 years ago. MRI showed a heterogeneous multiseptated large cystic mass within the mons pubis measuring 14 x 13.4 x 10.6 cm. Initial cytoreductive surgery revealed no evidence of tumor in the peritoneal cavity. The surgery was suboptimal due to severe adhesions to the symphysis pubis. The secondary cytoreductive surgery performed after six cycles of taxotere and carboplatin was optimal. Macroscopically, the tumor was a dusky pink-purple and contained a dense white-gray to light yellow gelatinous area. The tumor was a malignant cystic and glandular neoplasm. Immunohistochemical stains included CK7(+), CK5/6(-), EMA(+), Ber-Ep4(+), Calretinin(-), ER(-), and PR(-). CONCLUSION: Primary peritoneal clear cell adenocarcinoma arising from an abdominal scar associated with prior endometrioma/endometriosis surgery was first reported. PMID- 19219618 TI - A polymorphism in the promoter region of Ku70/XRCC6, associated with breast cancer risk and oestrogen exposure. AB - PURPOSE: Polymorphisms in double strand break repair genes could be involved in genetic breast cancer predisposition as enhanced chromosomal radiosensitivity is a hallmark for breast cancer. Previously, the c.-1310 C>G SNP, located in the Ku70 promoter, showed a significant odds ratio (OR) of 1.85 (P = 0.048) in sporadic, but not familial breast cancer patients, indicating that other factors besides genetic aptitude influence this association. As breast epithelium is exposed to endogenous oxidative stress through oestrogen exposure, the influence of hormone exposure was further examined. METHODS AND RESULTS: A significant OR (1.69, P = 0.017) was found for an enlarged patient population through PCR-RFLP assays in a case-control study in a Belgian population. After dividing the patient population according to oestrogen exposure, high and significant ORs were seen for patients with a longer oestrogen exposure (late age at menopause: OR = 1.96, P = 0.029). CONCLUSION: These results show that the variant allele of c. 1310 C>G, located in the Ku70 promoter, is a risk allele for breast cancer. Furthermore, the association of the c.-1310 C>G SNP with breast cancer risk was stronger in women with a long oestrogen exposure. PMID- 19219619 TI - Therapeutic effects of recombinant human endostatin adenovirus in a mouse model of malignant pleural effusion. AB - PURPOSE: Malignant pleural effusion (MPE) is a common clinical problem in patients with advanced cancer. Evidence suggests that tumor-mediated angiogenesis and enhanced vascular permeability in the pleural wall are due to high levels of vascular endothelial growth factor (VEGF), which plays an important role in the pathogenesis of MPE. The present study was designed to test whether the recombinant adenovirus-mediated delivery of human endostatin (Ad-hEndo), one of the potent inhibitors of angiogenesis, would inhibit the formation and progression of MPE. METHODS: We developed a novel mouse model of MPE by injecting Lewis lung carcinoma (LLC) cells directly into pleural cavity of C57BL/6 mice. To evaluate the therapeutic effects of endostatin in this MPE model, we injected the Ad-hEndo into the pleural cavity of MPE-bearing mice three times with the 3-day interval. RESULTS: We found that this treatment resulted in significant reduction in pleural effusion volume, the number of pleural tumor foci, microvessel density, and vascular permeability, while it significantly prolonged the survival time. In addition, VEGF level of MPE in the group administered with the Ad-hEndo was obviously decreased as compared with that in the two control groups administered with null-adenovirus (Ad-null) or normal saline. CONCLUSIONS: Our work provides a rationale for future studies toward evaluating the effectiveness of the adenovirus-based endostatin therapy for MPE. PMID- 19219620 TI - Intrinsically bent DNA sites in the Drosophila melanogaster third chromosome amplified domain. AB - Bent DNA sites promote the curvature of DNA in both eukaryotic and prokaryotic chromosomes. Here, we investigate the localization and structure of intrinsically bent DNA sites in the extensively characterized Drosophila melanogaster third chromosome DAFC-66D segment (Drosophila amplicon in the follicle cells). This region contains the amplification control element ACE3, which is a replication enhancer that acts in cis to activate the major replication origin ori-beta. Through both electrophoretic and in silico analysis, we have identified three major bent DNA sites in DAFC-66D. The bent DNA site (b1) is localized in the ACE3 element, whereas the other two bent DNA sites (b2 and b3) are localized in the ori-beta region. Four additional bent DNA sites were identified in the intron of the S18 gene and near the TATA box of the S15, S19, and S16 genes. The identification of DNA bent sites in genomic regions previously characterized as functionally relevant for DNA amplification further supports a function for DNA bent sites in DNA replication in eukaryotes. PMID- 19219621 TI - PHEX analysis in 118 pedigrees reveals new genetic clues in hypophosphatemic rickets. AB - Familial hypophosphatemic rickets is a rare disease, which is mostly transmitted as an X-linked dominant trait, and mutations on the phosphate regulating gene with homologies to endopeptidases on the X-chromosome (PHEX) gene are responsible for the disease in most familial cases. In this study we analyzed PHEX in a large cohort of 118 pedigrees representing 56 familial cases and 62 sporadic cases. The high-resolution melting curves technique was tested as a screening method, along with classical sequencing. PHEX mutations have been found in 87% of familial cases but also in 72% of sporadic cases. Missense mutations were found in 16 probands, two of which being associated with other PHEX mutations resulting into truncated proteins. By plotting missense mutations described so far on a 3D model of PHEX we observed that these mutations focus on two regions located in the inner part of the PHEX protein. Family members of 13 sporadic cases were analyzed and a PHEX mutation was detected in one of the apparently healthy mother. These results highlight the major role of PHEX in X-linked dominant hypophosphatemic rickets, and give new clues regarding the genetic analysis of the disease. A screening of the different family members should be mandatory when a PHEX mutation is assessed in a sporadic case and the search for another PHEX mutation should be systematically proceed when facing a missense mutation. PMID- 19219622 TI - On the effect of sharp rises in blood pressure in the Shah-Humphrey model for intracranial saccular aneurysms. AB - We consider the model originally proposed by Shah and Humphrey (J Biomech 32:593 599, 1999) for a class of intracranial saccular aneurysms and show that for constant pressure the addition of the viscoelastic term corresponding to the presence of cerebral spinal fluid outside the membrane, no matter how small, does ensure convergence to an equilibrium. Our arguments apply to a general equation of this type, and thus also hold for variations of this model such as that proposed by David and Humphrey (J Biomech 36:1143-1150, 2003). On the other hand, it is known that the presence of damping may destabilize periodic orbits of periodically forced systems or even prevent them from existing altogether. We present numerical simulations showing that for some forcing terms the high frequency oscillations do not die out with time, and a much more complex behaviour may emerge as a discontinuous forcing term is approached. The key point for this situation to occur is related to the derivative of the forcing term, supporting the hypothesis that sharper rises (or falls) in blood pressure may increase the risk of aneurysm rupture. PMID- 19219623 TI - Regulation of the extracellular antioxidant selenoprotein plasma glutathione peroxidase (GPx-3) in mammalian cells. AB - Plasma glutathione peroxidase (GPx-3) is a selenocysteine-containing extracellular antioxidant protein that catalyzes the reduction of hydrogen peroxide and lipid hydroperoxides. Selenoprotein expression involves the alternate recognition of a UGA codon as a selenocysteine codon and requires signals in the 3'-untranslated region (UTR), including a selenocysteine insertion sequence (SECIS), as well as specific translational cofactors. To ascertain regulatory determinants of GPx-3 expression and function, we generated recombinant GPx-3 (rGPX-3) constructs with various 3'-UTR, as well as a Sec73Cys mutant. In transfected Cos7 cells, the Sec73Cys mutant was expressed at higher levels than the wild type rGPx-3, although the wild type rGPx-3 had higher specific activity, similar to plasma purified GPx-3. A 3'-UTR with only the SECIS was insufficient for wild type rGPx-3 protein expression. Selenocompound supplementation and co-transfection with SECIS binding protein 2 increased wild type rGPx-3 expression. These results demonstrate the importance of translational mechanisms in GPx-3 expression. PMID- 19219624 TI - Activation of macrophage peroxisome proliferator-activated receptor-gamma by diclofenac results in the induction of cyclooxygenase-2 protein and the synthesis of anti-inflammatory cytokines. AB - Cyclooxygenase-2 (COX-2) is an inducible isoform of the COX family of enzymes central to the synthesis of pro-inflammatory prostaglandins. Induction of COX-2 is mediated by many endogenous and exogenous molecules that include pro inflammatory cytokines and bacterial lipopolysaccharide (LPS). It has been demonstrated that COX-2 can also be induced by diclofenac in cultured J774.2 macrophages. This induction was delayed compared to COX-2 induced by LPS and paracetamol selectively inhibited activity of this protein. The aim of the present study was to determine the transcription factor involved in the production of COX-2 after treatment of J774.2 cells with 500 microM diclofenac. Pre-treatment of cells with the peroxisome proliferator-activated receptor-gamma (PPAR-gamma) antagonists GW9662 (0.1-1 microM) or biphenol A Diglycidyl Ether (100-200 microM) resulted in reduction of the induction of COX-2 by diclofenac, but not by LPS. Induction of COX-2 by the PPAR-gamma agonist 15deoxyDelta(12,14)prostaglandin J(2) was also reduced when the cells were pre treated with the PPAR-gamma antagonists BADGE or GW9662. On the other hand, pre treatment of cells with the nuclear factor-kappa-B (NF-kappaB) Super-repressor IkappaBalpha (150-600 nM) reduced the induction of COX-2 by LPS, but not by diclofenac. We, therefore, have identified that PPAR-gamma activation is a requirement for COX-2 induction after diclofenac stimulation of J774.2 cells. These results along with the finding that treatment of J774.2 macrophages with diclofenac resulted in the release of the anti-inflammatory cytokines, interleukin-10 and transforming growth factor-beta suggest that the diclofenac induced COX-2 protein may possess anti-inflammatory actions. PMID- 19219625 TI - Betaine administration corrects ethanol-induced defective VLDL secretion. AB - Our previous studies, demonstrating ethanol-induced alterations in phosphatidylcholine (PC) synthesis via the phosphatidylethanolamine methyltransferase (PEMT) pathway, implicated a defect in very low-density lipoprotein (VLDL) secretion in the pathogenesis of hepatic steatosis. The objective of this study was to determine whether VLDL secretion was reduced by chronic ethanol consumption and whether betaine supplementation, that restores PEMT activity and prevents the development of alcoholic steatosis, could normalize VLDL secretion. The VLDL secretion in rats fed with control, ethanol and the betaine supplemented diets was determined using Triton WR-1339 to inhibit plasma VLDL metabolism. We observed reduced VLDL production rates in chronic alcohol-fed rats compared to control animals. Supplementation of betaine in the ethanol diet increased VLDL production rate to values significantly higher than those observed in the control diet-fed rats. To conclude, chronic ethanol consumption impairs PC generation via the PEMT pathway resulting in diminished VLDL secretion which contributes to the development of hepatic steatosis. By increasing PEMT-mediated PC generation, betaine results in increased fat export from the liver and attenuates the development of alcoholic fatty liver. PMID- 19219626 TI - Hyperthermia induced NFkappaB mediated apoptosis in normal human monocytes. AB - Conceptual approaches of heat-induced cytotoxic effects against tumor cells must address factors affecting therapeutic index, i.e., the relative toxicity for neoplastic versus normal tissues. Accordingly, we investigated the effect of hyperthermia treatment (HT) on the induction of DNA fragmentation, apoptosis, cell-cycle distribution, NFkappaB mRNA expression, DNA-binding activity, and phosphorylation of IkappaBalpha in the normal human Mono Mac 6 (MM6) cells. For HT, cells were exposed to 43 degrees C. FACS analysis showed a 48.5% increase in apoptosis, increased S-phase fraction, and reduced G2 phase fraction after 43 degrees C treatments. EMSA analysis showed a dose-dependent inhibition of NFkappaB DNA-binding activity after HT. This HT-mediated inhibition of NFkappaB was persistent even after 48 h. Immunoblotting analysis revealed dose-dependent inhibition of IkappaBalpha phosphorylation. Similarly, RPA analysis showed that HT persistently inhibits NFkappaB mRNA. These results demonstrate that apoptosis upon HT exposure of MM6 cells is regulated by IkappaBalpha phosphorylation mediated suppression of NFkappaB. PMID- 19219627 TI - Development of a substance abuse consultation and referral service in an academic medical center: challenges, achievements and dissemination. AB - Approximately 25% of US hospital beds are occupied by individuals with active substance use disorders (SUD). Acute medical hospitalization provides an opportunity to address SUDs and provide patient-centered intervention and referral for treatment. Nationally, some hospitals have developed substance abuse consultation departments to improve the care of hospitalized substance users. In this paper we describe the Addiction Psychiatry Service (APS) in a large urban hospital which provides bed-side SUD consultation, screening, intervention and referral to treatment. APS utilizes the multiple disciplines of psychology, social work and medicine to integrate substance abuse services throughout the hospital and educate future generations of medical and psychology trainees. We conclude with how the APS service is informing the development of similar programs in other academic departments within our hospital and best practice recommendations to further disseminate this service model. PMID- 19219628 TI - The problem of integrating behavioral health in the medical home and the questions it leads to. AB - Psychology and other behavioral health professions have amassed a broad empirical and clinical literature suggesting many medical presentations are best responded to with the addition of evidence based behavioral interventions. Despite this, psychology has not achieved a regular presence as part of medical practice. We suggest specific reasons for the current state of affairs including clinical, operational, societal labels, financial and training dimensions. Medical, psychological, administrative, and financial perspectives are reviewed. If the goals of health care system reform are to be reached then we must identify and challenge the current limitations of health care. This paper will identify the elements that need to be changed in order for psychology to be integrated into medicine rather than excluded from its policy, planning and operations. PMID- 19219629 TI - The prognostic impact of occult nodal metastasis in early breast carcinoma. AB - The clinical relevance of isolated tumor cell (ITC: 0.2-2.0 mm) in axillary lymph nodes (ALNs) remains unknown. The aim of this study was to determine their prognostic significance. A total of 295 patients considered as pN0 after routine histological assessment, were reevaluated with ten-level cytokeratin immunohistochemistry (IHC) and two-level hematoxylin-eosin sections. Survival rates, i.e. disease-free survival (DFS), distant disease-free survival (DDFS) and breast cancer specific survival (BCSS) were compared with those of reevaluated node-negative patients. A total of 84 patients (28%) had ITC/MM identified on IHC sections. ITC had no impact on survival at a median 8.2 years of follow-up, whereas MM showed a trend toward poorer DFS (P = 0.091, log rank) and DDFS (P = 0.066) and significantly reduced BCSS (P = 0.016). In multivariate analyses, detection of MM was an independent prognostic factor for DDFS (P = 0.025) and BCSS (P = 0.01) in adjuvant un-treated patients. Micrometastases (MMs) in axillary lymph nodes have prognostic impact. This was not found for ITC. This finding supports the use of systemic adjuvant therapy in patients with MM. PMID- 19219630 TI - The effect of transcutaneous electrical nerve stimulation on postmastectomy skin flap necrosis. AB - Transcutaneous electrical nerve stimulation (TENS) is primarily used for pain reduction and is thought to activate neural fibers modifying vascular resistance and local blood flow influencing flap necrosis. This study aimed to delineate the effects of TENS on the viability of skin flaps created during mastectomy in breast cancer patients. Patients treated with modified radical mastectomy were prospectively randomized to receive either TENS or no further local treatment postoperatively. High frequency (70 Hz) and low intensity (2 mA) TENS was applied and areas of flap ecchymosis and necrosis were measured after the completion of TENS application and compared between the two groups. Patients were also compared according to age, comorbid diseases, duration of anesthesia, flap area, amount of wound drainage and seroma, presence of wound infection and abscess. In addition, local and systemic complications related to TENS were recorded. About 173 patients with a median age of 49 (range 25-87) were included in the study. About 87 patients received TENS whereas 86 patients formed the control group. Patients' characteristics were comparable between the two groups. Number of patients with skin flap necrosis was higher in the control group compared to TENS group (P < 0.0001). Mean area of flap necrosis was significantly lower in TENS group (85.2 +/- 35.9) compared to control group (252.5 +/- 64.1; P = 0.024). Similarly, number of patients with skin flap ecchymosis was higher in the control group compared to TENS group (P = 0.002). However, although mean area of flap ecchymosis was lower in TENS group (105.5 +/- 49.8) compared to control group (172.9 +/- 49.9), this difference did not reach statistical significance (P = 0.34). In addition, there were no local or systemic complications related to TENS application in patients. TENS can be safely used to decrease the amount of skin flap necrosis after mastectomy in breast cancer patients without any additional complication. PMID- 19219631 TI - Sustained lower rates of HRT prescribing and breast cancer incidence in Australia since 2003. PMID- 19219632 TI - Docosahexaenoic acid ethyl ester enhances 6-hydroxydopamine-induced neuronal damage by induction of lipid peroxidation in mouse striatum. AB - Superoxide and hydroxyl radicals are implicated in the pathogenesis of Parkinson disease, and induction of lipid peroxidation is an important factor in progression of this disease. Docosahexaenoic acid (DHA) is a key component of the cell membrane, and its peroxidation is inducible due to the double-bond chemical structure. However, DHA has neuroprotective effects. In this study, we examined the effects of intraperitoneal injection (ipi) of DHA ethyl ester (DHA-Et) on 6 hydroxydopamine (6-OHDA)-induced dopamine (DA) reduction in the mouse striatum. DHA-Et ipi for 7 days before and 7 days after a single intracerebroventricular injection of 6-OHDA enhanced 6-OHDA-induced reduction of striatal DA level. On the other hand, ipi of DHA-Et for 7 days increased its concentration in the striatum. Co-injection of DHA-Et and 6-OHDA increased the levels of thiobarbituric acid-reactive substances (a marker of lipid peroxidation) in the striatum. Our results suggest that DHA-Et enhances 6-OHDA-induced DA depression by increasing lipid peroxidation, and that excessive use of DHA-Et may increase the susceptibility of Parkinson disease in animal model. PMID- 19219633 TI - Drug-resistant molecular mechanism of CRF01_AE HIV-1 protease due to V82F mutation. AB - Human immunodeficiency virus type 1 protease (HIV-1 PR) is one of the major targets of anti-AIDS drug discovery. The circulating recombinant form 01 A/E (CRF01_AE, abbreviated AE) subtype is one of the most common HIV-1 subtypes, which is infecting more humans and is expanding rapidly throughout the world. It is, therefore, necessary to develop inhibitors against subtype AE HIV-1 PR. In this work, we have performed computer simulation of subtype AE HIV-1 PR with the drugs lopinavir (LPV) and nelfinavir (NFV), and examined the mechanism of resistance of the V82F mutation of this protease against LPV both structurally and energetically. The V82F mutation at the active site results in a conformational change of 79's loop region and displacement of LPV from its proper binding site, and these changes lead to rotation of the side-chains of residues D25 and I50'. Consequently, the conformation of the binding cavity is deformed asymmetrically and some interactions between PR and LPV are destroyed. Additionally, by comparing the interactive mechanisms of LPV and NFV with HIV-1 PR we discovered that the presence of a dodecahydroisoquinoline ring at the P1' subsite, a [2-(2,6-dimethylphenoxy)acetyl]amino group at the P2' subsite, and an N2 atom at the P2 subsite could improve the binding affinity of the drug with AE HIV-1 PR. These findings are helpful for promising drug design. PMID- 19219634 TI - Spatial cluster analysis of early stage breast cancer: a method for public health practice using cancer registry data. AB - OBJECTIVES: Cancer registries are increasingly mapping residences of patients at time of diagnosis, however, an accepted protocol for spatial analysis of these data is lacking. We undertook a public health practice-research partnership to develop a strategy for detecting spatial clusters of early stage breast cancer using registry data. METHODS: Spatial patterns of early stage breast cancer throughout Michigan were analyzed comparing several scales of spatial support, and different clustering algorithms. RESULTS: Analyses relying on point data identified spatial clusters not detected using data aggregated into census block groups, census tracts, or legislative districts. Further, using point data, Cuzick-Edwards' nearest neighbor test identified clusters not detected by the SaTScan spatial scan statistic. Regression and simulation analyses lent credibility to these findings. CONCLUSIONS: In these cluster analyses of early stage breast cancer in Michigan, spatial analyses of point data are more sensitive than analyses relying on data aggregated into polygons, and the Cuzick Edwards' test is more sensitive than the SaTScan spatial scan statistic, with acceptable Type I error. Cuzick-Edwards' test also enables presentation of results in a manner easily communicated to public health practitioners. The approach outlined here should help cancer registries conduct and communicate results of geographic analyses. PMID- 19219635 TI - Second-hand smoke exposure in homes and in cars among Canadian youth: current prevalence, beliefs about exposure, and changes between 2004 and 2006. AB - PURPOSE: The present study examines second-hand smoke (SHS) exposure and the beliefs youth have about being exposed to SHS in their home and in cars and explores changes in exposure and beliefs over time. METHODS: Nationally representative data from the 2006 Youth Smoking Survey (YSS) were used to examine youth exposure to smoking and beliefs about smoking in the home and car among 71,003 Canadian youth in grades 5-12. Gender-specific logistic regression models were conducted to examine if being exposed to smoking at home or in the car were associated with the beliefs youth have about either smoking around kids at home or smoking around kids in cars. RESULTS: In 2006, 22.1% of youth in grades 5-12 were exposed to smoking in their home on a daily or almost daily basis and 28.1% were exposed to smoking while riding in a car at least once in the previous week. The majority of youth reported that they do not think smoking should be allowed around kids at home (88.3%) or in cars (88.4%). Youth exposed to smoking in the home or in cars reported missing substantially more days of school in the previous month because of their health. Among both male and female youth, being an ever smoker, living in a house where someone smokes inside daily, and having ridden in a car with someone who was smoking cigarettes in the past seven days were all associated with being more likely to report that smoking should not be allowed around kids at home or in cars. Compared to their male counterparts, female youth with at least one parent who smokes were more likely to report that smoking should not be allowed around kids at home or in cars. As rates of SHS exposure in the home and car decreased between 2004 and 2006, the prevalence of youth who reported that they do not think smoking should be allowed around kids at home or in cars also decreased over the same period of time. CONCLUSIONS: These results highlight that Canadian youth are frequently exposed to SHS in their homes and in cars despite the fact that the vast majority of youth do not think smoking should be allowed around kids in those locations. Considering the health and social consequences associated with SHS exposure, it may be a timely opportunity to move forward with programs and policies designed to prevent individuals from smoking around youth in these locations. PMID- 19219636 TI - A mWAP-hLF hybrid gene locus gave extremely high level expression of human lactoferrin in the milk of transgenic mice. AB - The production of pharmaceuticals by current mammary gland bioreactor techniques is limited by the low expression level of foreign proteins. We describe here a novel method that solves this problem. A successive three-step gap-repair strategy was developed to replace the genomic coding sequence in mouse whey acidic protein (mWAP) gene locus with that of human lactoferrin (hLF) precisely from the start code to the end code. A 50-kb mWAP-hLF hybrid gene locus was constructed, and corresponding transgenic mice were generated. An extremely high level expression of rhLF in the milk was demonstrated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blot. The expression level ranged from 16.7 to 29.8 g/l among five transgenic lines, as indicated by the ELISA assay. Importantly, the expressed rhLF maintained the same antibacterial activity as the native hLF. Our strategy can very likely also be used for the efficient expression of other valuable pharmaceutical proteins. PMID- 19219637 TI - Strategies for future histocompatible stem cell therapy. AB - Stem cell therapy based on the safe and unlimited self-renewal of human pluripotent stem cells is envisioned for future use in tissue or organ replacement after injury or disease. A gradual decline of regenerative capacity has been documented among the adult stem cell population in some body organs during the aging process. Recent progress in human somatic cell nuclear transfer and inducible pluripotent stem cell technologies has shown that patient-derived nuclei or somatic cells can be reprogrammed in vitro to become pluripotent stem cells, from which the three germ layer lineages can be generated, genetically identical to the recipient. Once differentiation protocols and culture conditions can be defined and optimized, patient-histocompatible pluripotent stem cells could be directed towards virtually every cell type in the human body. Harnessing this capability to enrich for given cells within a developmental lineage, would facilitate the transplantation of organ/tissue-specific adult stem cells or terminally differentiated somatic cells to improve the function of diseased organs or tissues in an individual. Here, we present an overview of various experimental cell therapy technologies based on the use of patient histocompatible stem cells, the pending issues needed to be dealt with before clinical trials can be initiated, evidence for the loss and/or aging of the stem cell pool and some of the possible uses of human pluripotent stem cell derivatives aimed at curing disease and improving health. PMID- 19219638 TI - Characterization of a novel 3-hydroxybutyrate dehydrogenase from Ralstonia pickettii T1. AB - We previously reported that the activities of two 3-hydroxybutyrate dehydrogenases (BDH1 and BDH2) were greatly influenced by culture conditions when Ralstonia pickettii T1, a strain growing on extracellular poly-3-hydroxybutyrate (PHB), was grown on different carbon sources such as 3HB and succinate. In this study, knockout mutants of bdh1 or bdh2 were constructed and characterized under different culture conditions. In addition, a novel BDH (BDH3) was found in bdh2 mutants, and bdh3 was cloned. Apparent kinetic parameters for the substrates of BDH3 indicated that the enzyme is suitable for the oxidation reaction of 3 hydroxybutyrate (3HB) to acetoacetate. In Western blotting, it was clear that BDH3 is produced only in cells grown on 3HB or PHB as a carbon source, while BDH1 and BDH2 are produced in cells grown on various carbon sources such as sugars, amino acids, organic acids, 3HB, and PHB. Both the bdh1 and bdh2 mutants lagged behind the wild type in growth rates when the cells were cultured with 3HB, citrate, succinate, or nutrient broth. A test of sensitivity to diamide as an oxidative stress revealed that the lack of BDH1 or BDH2 caused a decline in the capacity to neutralize the stress. These results suggested that BDH1 and BDH2 are needed to regulate the cytoplasmic redox state as well as to utilize 3HB, while BDH3 is specialized to utilize 3HB. The expression of bdh3 may be coordinately regulated with a gene encoding putative 3HB permease. PMID- 19219639 TI - Microarray analysis of gene expression in seeds of Brassica napus planted in Nanjing (altitude: 8.9 m), Xining (altitude: 2261.2 m) and Lhasa (altitude: 3658 m) with different oil content. AB - The regulation of seed oil synthesis in rapeseed is largely unknown. In this study, Arabidopsis microarray was used to analyze the gene differential expression of the immature seeds 30 days after flowering of a high oil Brassica napus, H105, whose oil content was 46.04 +/- 1.42, 53.94 +/- 1.35 and 53.09 +/- 1.35% when planted in Nanjing (altitude: 8.9 m), Xining (altitude: 2261.2 m) and Lhasa (altitude: 3658 m), respectively. Transcript levels of 363 genes and 421 genes were altered twofold or more for H105 planted in Xining and Lhasa compared to that in Nanjing, respectively. Together, there were 53 common up-regulated and 42 common down-regulated expression transcripts shared by H105 planted in Xining and Lhasa compared to that in Nanjing. Some important genes, such as sucrose synthase, pyruvate kinase and 6-phosphogluconate dehydrogenase which related to sugar metabolism were identified common up-regulated in higher oil content H105. These results revealed the expressional disciplinarian of correlative genes, and provided important information of the molecular genetic mechanism of oil content difference of rapeseed. In addition, these differential expression genes could be suitable as targets for genetic improvement of seed oil content. PMID- 19219640 TI - The prevalence of factor V Leiden, prothrombin G20210A and methylenetetrahydrofolate reductase polymorphism C677T among G6PD deficient individuals from Western Iran. AB - It has been suggested that the allele frequency of thrombophilic mutations is affected by glucose-6-phosphate dehydrogenase (G6PD) deficiency. The prevalence of thrombophilic mutations were studied in sixty G6PD deficient individuals including 57 males and three females with the mean age of 15 +/- 3.08 and 110 age and sex matched healthy individuals consisted of 95 males and 15 females with the mean age of 16.19 +/- 2.17 from the Kermanshah Province of Iran. Using a combination of PCR-RFLP technique, single strand conformation polymorphism (SSCP) analysis and DNA sequencing polymorphic G6PD mutations were identified. The factor V Leiden, prothrombin G20210A and methylenetetrahydrofolate reductase (MTHFR) C677T were detected by PCR-RFLP method using MnlI, HindIII and HinfI restriction enzymes, respectively. Three mutations, G6PD Mediterranean, G6PD Chatham and G6PD Cosenza were identified in 60 G6PD deficient individuals with highest prevalence of G6PD Mediterranean (91.6%). In G6PD deficient individuals the prevalence of factor V Leiden tended to be higher (5%) compared to healthy individuals (2.7%). The prevalence of prothrombin G20210A mutation in G6PD deficient individuals was 1.7%. However, in normal subjects the prevalence of this mutation was 2.7%. The frequency of T allele in G6PD deficient individuals were insignificantly higher (29.16%) than those in healthy individuals (26.8%). Our finding indicates that the prevalence of factor V Leiden, prothrombin G20210A and MTHFR C677T in G6PD deficient individuals is not statistically different compared to normal subjects and G6PD deficiency is not associated with these thrombophilic mutations in Western Iran. PMID- 19219641 TI - The hermeneutic challenge of genetic engineering: Habermas and the transhumanists. AB - The purpose of this paper is to explore the impact that developments in transhumanist technologies may have upon human cultures (and thus upon the lifeworld), and to do so by exploring a potential debate between Habermas and the transhumanists. Transhumanists, such as Nick Bostrom, typically see the potential in genetic and other technologies for positively expanding and transcending human nature. In contrast, Habermas is a representative of those who are fearful of this technology, suggesting that it will compound the deleterious effects of the colonisation of the lifeworld, further constraining human autonomy and undermining the meaningfulness of the lifeworld by expanding the technological control and manipulation of humanity. It will be argued that these opposed positions are grounded in fundamentally different understandings of the consequences of scientific and technological advance. On one level, the transhumanists remain confident that the lifeworld has within it the resources necessary to find meaning and purpose in a society deeply infused by genetic technology. Habermas disagrees. On another level, the difference is articulated by Horkheimer and Adorno in Dialectic of Enlightenment, primarily by challenging what may be understood as a Baconian faith in science as a project for the domination of nature (where nature is an infinitely malleable material, to be dominated and shaped, without adverse consequences, purely for the purposes of human survival). While the transhumanists broadly embrace this faith, Habermas returns to something akin to Horkheimer and Adorno's pessimistic scepticism. PMID- 19219642 TI - Epidemiology and causation. AB - Epidemiologists' discussions on causation are not always very enlightening with regard to the notion of 'cause' in epidemiology. Epidemiologists rightly work from a science-based approach to causation in epidemiology, but largely disagree about the matter. Disagreement may be partly due to confusion of the question of useful concepts for causal inference in epidemiological practice with the question of the metaphysical presuppositions of causal concepts used in epidemiology. In other words, epidemiologists seem to confuse the practical results of epidemiological research at the population level with the metaphysical views about the reality of disease causation at the individual level in their writings on causation. PMID- 19219643 TI - Correlation between synovial blood flow signals and serum vascular endothelial growth factor levels in patients with refractory rheumatoid arthritis. AB - The objective of the study is to examine the relationship between synovial blood flow signals and vascular endothelial growth factor (VEGF) involved in angiogenesis by Doppler ultrasound. Twenty-one patients meeting the diagnostic criteria of the American College of Rheumatology (ACR) were enrolled in this study. Doppler ultrasound signals of blood flow in the wrist synovial membrane were measured and classified into three grades: grade 1 = no flow; grade 2 = mild flow; grade 3 = intense flow. A significant correlation was observed between blood flow signals in the wrist synovial membrane and serum VEGF levels (r = 0.5681, P = 0.0072). These results suggest that the measurement of Doppler ultrasound signals of blood flow in the wrist synovial membrane is useful in the evaluation of angiogenesis. PMID- 19219644 TI - Morphological study, by teasing examination, of the communication from the musculocutaneous to median nerves. AB - Nine specimens with communications from the musculocutaneous to the median nerves were investigated, by teasing examination, from their origins in the brachial plexus to their final destinations in nerve fibers. The nerve fibers of the communications were derived from the sixth and seventh cervical ventral rami of the spinal nerves. The distributions of the nerve fibers of the communications were divided into four types. In Type A, the nerve fibers reached the thenar muscles and the lateral digital nerves. In Type B, they reached the pronator teres or flexor carpi radialis muscles in addition to Type A. In Type C, they reached the anterior interosseous nerve area in addition to Type B. Finally in Type D, they also reached the distal muscle belly of the index of the flexor digitorum superficialis. It was revealed that there was a definite rule in the distribution of the nerve fibers in the communications from the musculocutaneous to median nerves. The area of the distributions was expanded in order from the thenar muscles to the flexor muscles of the forearm. The results in this study are useful for proper diagnosis and treatment of the peripheral nerve injuries involving the musculocutaneous and median nerves. PMID- 19219645 TI - Morphological study of the communication between the musculocutaneous and median nerves. AB - We investigated the incidence of each pattern of the communications between the musculocutaneous and median nerves and discussed its morphological significance. The communications between both nerves were seen in 188 (41.5%) of 453 specimens. They were classified into five regular patterns according to their directions and formations. The incidence of the communication was significantly higher in those with excessive heads of biceps brachii than in those without them (p < 0.05). As a result, we established a classification of the regular pattern of the communications and revealed that the musculocutaneous and median nerves had the possibility of forming plexuses in the humeral area. The musculocutaneous nerves are regarded as the bundles that arise from the median nerve with the lateral cutaneous nerve of forearm and the branches to the biceps brachii and brachialis muscles. PMID- 19219646 TI - Chemical, biochemical, and biological impact of untreated domestic sewage water use on Vertisol and its consequences on wheat (Triticum aestivum) productivity. AB - In the peri-urban areas of central India, sewage water is a valuable resource for agricultural production. In this study, impact of domestic sewage water irrigation for 5 years on Vertisol with no previous history of sewage irrigation was investigated in an ongoing field experiment at Bhopal (India) under subtropical monsoon type climate. The wheat (Triticum aestivum) crop was grown during post-rainy winter season with 30 cm of irrigation (groundwater or sewage water) and four nutrient treatments (T(1), 0; T(2), 100%; T(3), 50%; and T(4), 50% of general recommended doses of NPK + FYM at 10 Mg/ha). Results showed that sewage irrigation of about 150 cm over a period of 5 years resulted significant increases in salinity as well as available fractions of N, P, K, and micronutrients, viz., Zn, Fe, and Mn in soils. Carbon and phosphorus applied through sewage water were accumulated more in subsoil layer compared to topmost plough layer. Soil microbiological activity, as indicated by soil respiration, microbial biomass C, as well as dehydrogenase enzyme activity was higher in sewage water-irrigated soils. There was also significant increase in fungal and actinomycetes as well as total coliform population in such soils. Nutrients supplied through sewage water were not able to raise the productivity of wheat to the level that obtained through fertilizers at the recommended level which indicated that additional nutrients through fertilizers are required to obtain higher productivity of wheat under sewage farming. Protein and Zn content in wheat grains were more when the crop was grown with sewage irrigation. Overall results show that except for increase in coliform population, short duration (5 years) of municipal sewage water irrigation did not have any appreciable harmful effect on soil quality as well as crop productivity; rather, it proved beneficial in improving soil fertility, wheat productivity, and produce quality. PMID- 19219647 TI - Adolescent suicidal ideation: a comparison of incarcerated and school-based samples. AB - BACKGROUND: The aim of this study was to investigate suicidal ideations and associated psychopathology in two groups of adolescents, a sample of detained youth and a general population sample. In both groups the comparisons of mental health characteristics between suicidal ideators and non-suicidal youth were conducted separately for girls and boys. METHODS: The study sample consisted of 290 delinquent adolescents [228 boys and 62 girls] from three Flemish juvenile detention centers and 1,548 adolescents [811 boys and 737 girls] from an age matched school-based sample. Both groups were administered the Social and Health Assessment [SAHA], a self-report survey investigating levels of psychopathology [internalizing and externalizing] and risk-taking behavior. RESULTS: Suicidal ideations during the past year were reported by 21.5% of detained males, compared to 6.7% in the general population. In females, 58.1% of detained individuals reported suicidal thoughts during the past year, compared to 14.4% of the general population. In girls and boys from the general population, both internalizing and externalizing problems were higher in suicidal ideators than in non-suicidal youth, while in the detention group mainly internalizing problems were higher in suicidal ideators. When comparing detention suicidal ideators with those from the general population, male suicidal ideators scored higher on delinquency, while detained female suicidal ideators also scored higher on posttraumatic stress, but lower on prosocial beliefs. LIMITATIONS: Information used in this study was solely based on self-report measures only and limited to Flemish adolescents. CONCLUSION: Since suicidal ideation is a frequent problem in detained youth, adequate recognition and treatment seems clinically relevant. While both internalizing and externalizing psychopathology may be an indicator of suicidal ideation in the general population, internalizing problems may be the main clinical predictor in detained youth. PMID- 19219648 TI - Influence of parent's eating attitudes on eating disorders in school adolescents. AB - OBJECTIVE: To investigate the relationship between parents' cognitive and behavioural dimensions and the risk of eating disorders (ED) in non-clinical adolescents. METHODS: From an initial sample of 1,336 boys and girls with a mean age of 11.37, a total of 258 subjects were selected either as being at risk of ED or as controls. These subjects and their parents comprised the sample at T1 and were followed-up 2 years later (T2). We examined disordered eating attitudes, body dissatisfaction (BD), body mass index (BMI) and ED diagnoses in the adolescents at two points in time. We also used the Eating Disorder Inventory-2 to examine the disordered eating attitudes of their parents at T1. RESULTS: The mother's BD, drive for thinness (DT), ineffectiveness and interoceptive awareness, and the father's DT and perfectionism were related to long-term ED. Logistic regression showed that predictors of ED were being female, mother's DT and social insecurity, and adolescent's BD. The BMI was not a predictor in this model. Father's perfectionism was a risk factor of ED. CONCLUSION: Specific cognitive and behavioural dimensions of ED in both parents can influence the development of an ED in early adolescents. This should be taken into account in the prevention and family oriented treatment of ED. PMID- 19219649 TI - Block design reconstruction skills: not a good candidate for an endophenotypic marker in autism research. AB - Superior performance on block design tasks is reported in autistic individuals, although it is not consistently found in high-functioning individuals or individuals with Asperger Syndrome. It is assumed to reflect weak central coherence: an underlying cognitive deficit, which might also be part of the genetic makeup of the disorder. We assessed block design reconstruction skills in high-functioning individuals with autism spectrum disorders (ASD) from multi incidence families and in their parents. Performance was compared to relevant matched control groups. We used a task that was assumed to be highly sensitive to subtle performance differences. We did not find individuals with ASD to be significantly faster on this task than the matched control group, not even when the difference between reconstruction time of segmented and pre-segmented designs was compared. However, we found individuals with ASD to make fewer errors during the process of reconstruction which might indicate some dexterity in mental segmentation. However, parents of individuals with ASD did not perform better on the task than control parents. Therefore, based on our data, we conclude that mental segmentation ability as measured with a block design reconstruction task is not a neurocognitive marker or endophenotype useful in genetic studies. PMID- 19219650 TI - Statins and breast cancer: may matrix metalloproteinase be the missing link. AB - Statins (3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors), cholesterol lowering agents widely prescribed for cardiovascular health, have been shown to exert several pleiotropic effects. Although some studies reported that statins have no effects on malignancies of any kind, results of several epidemiologic and in vitro studies highlighted that statins exert anticancer activity in various cell types, showing that long-term therapy inhibits the incidence and/or progression of some human tumours. In particular, in the present overview we focused the attention on a neglected aspect of the pleiotropic functions of some lipophilic statins, suggesting that the possible mechanism of matrix metalloproteinase downregulation arises from prolonged lowering of circulating cholesterol. Our hypothesis may explain the literary findings about the phenomenon of switching of breast cancer phenotypes by statins, shedding the basis of future epidemiologic and basic science studies about the role of circulating and/or tumor-resident cholesterol in the initiation and progression of breast cancer. PMID- 19219651 TI - Radiation recall dermatitis after pre-sensitization with pegylated liposomal doxorubicin. AB - Radiation recall dermatitis (RRD) is a rare cutaneous reaction occurring within a previously irradiated field, precipitated by certain drugs. We report a case of RRD occurring after pre-sensitization with pegylated liposomal doxorubicin (PLD) in a woman with Stage IV breast cancer. The RRD occurred in one of the patient's four previous radiotherapy fields. We discuss the time/dose factors of radiation exposure and measure the corresponding skin dose. In our case the radiation dose was low and below previously reported thresholds, and illustrates that there is a more complex interaction between the radiotherapy and the trigger agent than has previously been considered. PMID- 19219652 TI - The effect of TGF-beta2 on MMP-2 production and activity in highly metastatic human bladder carcinoma cell line 5637. AB - Transforming growth factor-beta (TGF-beta) superfamily regulates matrix metalloproteinases (MMP), which intrinsically regulate various cell behaviors leading to metastasis. We investigated the effect of TGF-beta(2) on MMP-2 regulation in human bladder carcinoma cell line 5637. Zymography, ELISA, and real time polymerase chain reaction revealed that TGF-beta(2) stimulated MMP-2 production, but the transcription of its gene remained unchanged. Wortmannin could not inhibit MMP-2 secretion and activity and conversely the amount of the protein and its enzymatic activity were increased. These data suggest that TGF beta(2) increased MMP-2 at the posttranscriptional level and this upregulation was independent of phosphatidylinositol 3-kinase signaling pathway. PMID- 19219653 TI - Expression profiling of nucleotide metabolism-related genes in human breast cancer cells after treatment with 5-fluorouracil. AB - 5-Fluorouracil (5-FU) is one of the most widely used drugs for treatment of cancers, including breast cancer that exhibits its anticancer activity by inhibiting DNA synthesis and also incorporated into DNA and RNA. The objective of this investigation was to find out the total nucleotide metabolism genes regulated by 5-FU in breast cancer cell line. The breast cancer cell line MCF-7 was treated with the drug 5-FU. To analyze the expression of genes, we have conducted the experiment using 1.7 k and 19k human microarray slide and confirmed the expression of genes by semiquantitative reverse transcription-polymerase chain reaction. The expression of 44 genes involved in the nucleotide metabolism pathway was quantified. Of these 44 genes analyzed, transcription of 6 genes were upregulated and 9 genes were downregulated. Earlier studies revealed that the transcription of genes for key enzymes like thymidylate synthase, thymidine kinase, and dihydropyrimidine dehydrogenase are regulated by 5-FU. This study identified some novel genes like thioredoxin reductase, ectonucleotide triphosphate dephosphorylase, and CTP synthase are regulated by 5-FU. The data also reveal large-scale perturbation in transcription of genes not involved directly in the known mechanism of action of 5-FU. PMID- 19219654 TI - DNA copy number variation and gene expression analyses reveal the implication of specific oncogenes and genes in GBM. AB - To understand the pathogenesis of glioblastoma multiforme (GBM) we used high resolution comparative genomic hybridization arrays and gene expression microarrays to identify DNA copy number alterations and gene expression changes in comparable sets of GBM samples. Gains were detected at chromosomes 1, 2, 7, 9, 12, 19, and 20 and losses at 6, 9, and 10. Gene expression analyses identified specific genes overexpressed in GBM mapping at amplified chromosomal regions. Among these genes we found genes involved in angiogenesis, extracellular matrix remodeling and several oncogenes. DNA copy number analysis along with gene expression profiles provides a powerful strategy to understand tumor progression and identification of genes involved in GBM pathogenesis. PMID- 19219655 TI - Tissue factor and VEGF expression in prostate carcinoma: a tissue microarray study. AB - Tissue factor (TF) is the principal physiologic initiator of coagulation. It also plays an important role in tumor growth and metastasis possibly by contributing to angiogenesis. We evaluated the expression of TF in benign and malignant prostate tissue and correlated it with the expression of the pro-angiogenic protein, vascular endothelial growth factor (VEGF). We used a tissue microarray (TMA) constructed from 80 archival prostatectomy specimens. Core samples were collected from benign prostate tissue (BP) (n= 77), high-grade prostatic intraepithelial neoplasia (PIN) (n= 26), and carcinoma (PCa) (n= 93). TMA sections were stained with an immunopurified polyclonal TF antibody and a rabbit polyclonal VEGF. Two pathologists manually scored staining in epithelial cells using the German Immunoreactive Score. Positive staining for TF was seen predominantly in PCa with rare positive glands in BP and PIN. TF expression was significantly lower in BP versus PCa specimens (p< .001) and in PIN versus PCa specimens (p< .001). Positive staining for VEGF was seen in PCa, BP, and PIN. Rates of VEGF expression were also significantly lower in BP versus PCa specimens (p= .003) but not in PCa versus PIN (p= .430). The majority of PCa samples positive for TF were also positive for VEGF (p< .001). Our findings reinforce the link between angiogenesis and TF expression in PCa. We suggest further exploration of TF-mediated pathways leading to increased tumor aggressiveness in PCa, and the possible use of anti-TF agents in PCa. PMID- 19219656 TI - Cross-talk between Notch and EGFR signaling in human breast cancer cells. AB - Notch and epidermal growth factor receptor (EGFR) signaling play critical roles in cell proliferation, differentiation, and apoptosis, and thereby may contribute to the development of breast cancer. We constitutively overexpressed active Notch1 in human breast cancer cells to explore the consequences of Notch1 signaling on cell growth and to investigate the underlying molecular mechanisms. We found that EGFR expression was increased. Then, using EGFR inhibitor, we found it exhibited an inhibitory role on human breast cancer cells. Overexpression of Notch1 could reverse EGFR inhibitor-induced cell toxicity, suggesting that Notch and EGFR signaling may be positively cross-linked in human breast cancer. PMID- 19219657 TI - Predicting internet pornography use and arousal: the role of individual difference variables. AB - This study considers the relation between a number of theoretically relevant individual difference variables and individuals' online pornography use and arousal patterns. In doing so, an attempt is also made to determine whether self reports of arousal can be collapsed into meaningful empirically derived content groupings. An exploratory factor analysis produces 3 factors for men: standard fare, specialized, and male-focused; and two factors for women: standard fare and specialized. Findings indicate that sexual disposition is a strong predictor of standard fare use and arousal for both genders. Men and women higher in psychopathy were more likely to use all forms of content. For those high in psychopathy, however, men were only slightly, and women not at all, more likely to find standard fare content arousing. Results are discussed in terms of their potential value for understanding an often overlooked first step in the process of pornography consumption. PMID- 19219658 TI - Influence of spirituality and religiousness on substance misuse in patients with schizophrenia or schizo-affective disorder. AB - Substance misuse represents a major issue in the treatment of schizophrenia patients. Spirituality and religiousness have been shown to reduce substance misuse and to foster recovery among substance misusers in the general population. One hundred and fifteen stabilized outpatients with schizophrenia (mean age 39; 70% male) were selected in 2004 for an interview about religious coping. Religious involvement was significantly inversely correlated to substance use and abuse. A content analysis showed that religion may play a protective role toward substance misuse in 14% of the total sample, especially for patients who had stopped substance misuse (42%). It played a negative role in 3% of cases. Religion may play a role in the recovery of schizophrenia patients with substance misuse comorbidity. PMID- 19219659 TI - Cautionary guidelines for the use of opioids in chronic pain. AB - Guidelines for the use of opioids in the treatment of chronic pain have recently been proposed by the American Academy of Addiction Psychiatry. Older guidelines proposed by American pain organizations had the effect of liberalizing opioid prescription. In recent years, dramatic increases in prescribed opioids have been followed by equally dramatic rises in morbidity and mortality from prescription opioids. In addition, new research has increased knowledge of the long-term effects of opioids. These new guidelines propose increased caution in regard to opioid prescription for chronic pain. PMID- 19219660 TI - Opioid deaths in rural Virginia: a description of the high prevalence of accidental fatalities involving prescribed medications. AB - In rural Virginia, drug overdose deaths increased 300% from 1997 to 2003. Polydrug deaths predominate (57.9%) in this review of 893 medical examiner cases. Prescription opioids (74.0%), antidepressants (49.0%), and benzodiazepines (39.3%) were more prevalent than illicit drugs. Two-thirds of decedents were 35 54 years old; 37% were female. When compared to western Virginia metropolitan cases, polydrug abuse was more common, specific medication combinations were found, the death rate per population was higher, and fewer illicit drugs were detected. These rural prescription overdose deaths differ from urban illicit drug deaths, suggesting the need for different strategies in prevention, treatment, and intervention by clinicians and policymakers. PMID- 19219661 TI - Treatment challenges associated with comorbid substance use and posttraumatic stress disorder: clinicians' perspectives. AB - A significant proportion of individuals with substance use disorders (SUDs) meet criteria for comorbid posttraumatic stress disorder (PTSD). This comorbidity confers a more complicated clinical presentation that carries with it formidable treatment challenges for practitioners. The current study examined sources of difficulty and gratification among clinicians (N = 423) from four national organizations who completed an anonymous questionnaire. As expected, the findings revealed that comorbid SUD/PTSD was rated as significantly more difficult to treat than either disorder alone. The most common challenges associated with treating SUD/PTSD patients included knowing how to best prioritize and integrate treatment components, patient self-destructiveness and severe symptomatology, and helping patients abstain from substance use. The findings increase understanding of SUD/PTSD treatment challenges, and may be useful for enhancing therapist training programs, supervision effectiveness, and designing optimal SUD/PTSD interventions. PMID- 19219662 TI - The inclusion of women and minorities in smoking cessation clinical trials: a systematic review. AB - This study assesses the impact of the 1993 NIH Revitalization Act on the inclusion and subgroup analysis of women and minorities in trials of FDA-approved smoking cessation pharmacotherapy. Female representation, while commensurate with population levels, declined significantly for trials that began recruitment after 1993(M = 47.2% vs. M = 53.9%), and fewer than half reported analyses by gender. Minorities continued to be under-represented in later trials; however, significant improvement in representation (M = 16.1% vs. M = 10%) and analysis by race occurred. Industry-sponsored studies had lower minority representation than NIH funded studies. Recommendations are offered to improve subgroup analyses and minority inclusion. PMID- 19219663 TI - Alcohol and injury in the United States general population: a risk function analysis from the 2005 National Alcohol Survey. AB - A risk function analysis of data is reported from the 2005 National Alcohol Survey on average daily volume of alcohol and frequency of consuming five or more drinks per day with reporting any injury during the preceding year, an injury that was treated, or an injury that was treated in the emergency room (ER). Data suggest the association of alcohol and injury based on ER studies is not representative of all injuries, and should be taken into account in deriving estimates of risk of injury related to alcohol and alcohol-attributable fractions based on ER studies. PMID- 19219664 TI - Prediction of cannabis use disorder between boyhood and young adulthood: clarifying the phenotype and environtype. AB - Employing a prospective paradigm, this investigation derived the childhood phenotype and the environtype associated with risk for cannabis use disorder. Two hundred and sixteen boys were evaluated between age 10-12 on a comprehensive protocol using self, mother, and teacher reports and followed-up at ages 19 and 22 to determine the presence of cannabis use disorder. The Transmissible Liability Index (TLI) and Non-Transmissible Liability Index (NTLI) were derived using item response theory. Logistic regression was conducted to evaluate the accuracy of the indexes, singly and in combination, to predict cannabis use disorder. The TLI and NTLI together predicted with 70% and 75% accuracy cannabis use disorder manifest by age 19 and age 22. Sensitivity was 75% at both ages 19 and 22, whereas specificity was respectively 51% and 64%. The findings pertaining to sensitivity indicate that SUD risk for cannabis use disorder can be screened in childhood; however, the specificity scores demonstrate that a low score on the TLI does not inevitably portend a good prognosis up to 10 years later. PMID- 19219665 TI - Olanzapine in cocaine dependence: a double-blind, placebo-controlled trial. AB - Preclinical and uncontrolled human studies have suggested the possible efficacy of second-generation antipsychotics, particularly olanzapine, in treating cocaine dependence. We conducted a randomized, double-blind, placebo-controlled trial in which 48 cocaine-dependent subjects received olanzapine or identical-appearing placebo for 16 weeks. The primary outcome measure was the proportion of cocaine negative weekly urine screens during treatment. Secondary measures included scores on a Craving Questionnaire, Addiction Severity Index subscales, and extrapyramidal symptom scales. Olanzapine and placebo did not differ on any outcome measure. Both olanzapine and placebo subjects frequently reported side effects, but no unexpected ones. We conclude that olanzapine appears ineffective for cocaine dependence. PMID- 19219666 TI - A preliminary trial: double-blind comparison of nefazodone, bupropion-SR, and placebo in the treatment of cannabis dependence. AB - The present study investigated the efficacy of nefazodone and bupropion-sustained release for treating cannabis dependence. A double-blind, placebo-controlled, piggy back design was employed to assess if nefazodone and bupropion-sustained release increased the probability of abstinence from cannabis and reduced the severity of cannabis dependence and cannabis withdrawal symptoms during a 13-week outpatient treatment program. One-hundred and six participants (Mean = 32 years; females n = 25) were randomized to one of three medication conditions (nefazodone, bupropion-sustained release, or placebo) and participated in a weekly, individually based coping skills therapy program. Results indicated an increased probability of achieving abstinence over the course of treatment and a decrease in the severity of cannabis dependence and the withdrawal symptom of irritability. There were no significant effects demonstrated for nefazodone and bupropion-sustained release on cannabis use or cannabis withdrawal symptoms. The results indicate nefazodone and bupropion-sustained release may have limited efficacy in treating cannabis dependence. PMID- 19219668 TI - Low plasma levels of docosahexaenoic acid are associated with an increased relapse vulnerability in substance abusers. AB - Low levels of some polyunsaturated fatty acids (PUFAs) could influence behaviors leading to the abuse of substances through their actions on serotonergic and dopaminergic mechanisms. Because substance abusers tend to have poor dietary habits, the possibility that a deficient intake of n-3 PUFAs, available from dietary sources only, and subsequent low n-3 plasma levels would predict their relapse rates was explored. Thirty-five patients admitted to substance abuse clinics were enrolled and followed for one year. Dietary questionnaires and blood samples were collected at baseline and on a quarterly basis, and relapse rates monitored on a monthly basis. Six patients dropped out shortly after study entry, 11 relapsed in the course of the study and dropped out, 7 relapsed but completed the study, and 11 did not relapse and completed the study. Non-relapsers were found to have significantly higher levels of docosahexaenoic acid (DHA) calculated as microg/ml and % TFA, when compared to relapsers (p = .031 and p = .010, respectively) and to relapsers and non-completers combined (p = .014 and p = .009, respectively). These pilot data suggest, but do not prove, the existence of a relationship between low levels of DHA and relapse vulnerability in some individuals who abuse substances. The study of the efficacy of n-3 supplements or of dietary modifications on relapse appears warranted. PMID- 19219667 TI - Sexual risk behavior associated with co-administration of methamphetamine and other drugs in a sample of HIV-positive men who have sex with men. AB - This study examined the association between sexual risk behavior and co administration of methamphetamine with other drugs in a sample of 341 HIV positive MSM. Those who reported methamphetamine co-administration in the past two months (65%) reported significantly more unprotected anal and oral sex and a greater number of casual, anonymous, and paid sex partners in this timeframe compared to men who used methamphetamine alone. Two primary patterns of co administration were identified: 1) drug combinations motivated by sexual performance and enhancement (eg, methamphetamine, poppers, sildenafil); and 2) "party drug" combinations (eg, methamphetamine, GHB, ketamine). Implications for further research and possible applications to risk-reduction interventions are discussed. PMID- 19219670 TI - A comparison of substance use disorder severity and course in American Indian male and female veterans. AB - The purpose of this analysis was to compare substance use disorder (SUD) diagnoses, severity, comorbidity, and course in 362 American Indian veterans. The sample was drawn from communities in the north central and southwestern areas of the United States, structured to over-sample women and to include half-rural/half urban residents. Instruments used in the study included current demography, military history, the Diagnostic Interview Schedule/Quick Version, Michigan Alcoholism Screening Test modified to include alcohol and drugs (MAST/AD), Brief Symptom Inventory, Posttraumatic Checklist, and a treatment algorithm. Univariate analyses showed that women had lower scores on the MAST/AD, reported lower symptom levels on the Posttraumatic Checklist, and were less apt to use VA mental health services, but were more willing to seek mental health treatment if needed (with probabilities of .01 to .001). At borderline probabilities (.02 to .05), women were younger and had more education, whereas men had more drug-related diagnoses and more combat exposure. On binary logistic regression, women were less apt to have a drug diagnosis and had lower MAST/AD scores; the other differences were not significant. Female American Indian veterans with SUD manifested many of the same gender-related differences as women in the population at large, but with some exceptions (eg, comorbidity). Trauma, PTSD, and continuing posttraumatic symptoms occurred frequently in both male and female veterans of American Indian heritage. VA facilities should out-reach to American Indian women, who report a willingness to seek mental health services but may avoid VA care. PMID- 19219671 TI - Anxiety sensitivity as a moderator of the association between smoking rate and panic-relevant symptoms among a community sample of middle-aged adult daily smokers. AB - The aim of the present investigation was to evaluate the moderating role of anxiety sensitivity (AS) in the relation between smoking rate and panic vulnerability variables among a community-based sample of adults. Results indicated that the interaction between AS and smoking rate significantly predicted anxious arousal, agoraphobic avoidance, and anticipatory anxiety. Specifically, participants who reported higher levels of AS and heavier smoking rates reported the highest levels of panic vulnerability. These data suggest that this combination of high AS and heavier smoking is particularly problematic in regard to panic symptoms. PMID- 19219669 TI - Predictors of treatment outcome in outpatient cocaine and alcohol dependence treatment. AB - We examined the ability of several baseline variables to predict treatment outcome in a pharmacotherapy trial that included 164 participants who were both cocaine- and alcohol-dependent and were selected for a randomized, double-blind, placebo-controlled study. Predictor variables included results from the baseline Addiction Severity Index (ASI), initial Urine Drug Screen results, cocaine and alcohol craving and cocaine and alcohol withdrawal symptoms at the start of treatment. Successful treatment was defined as four continuous weeks of self reported cocaine abstinence verified by urine drug screens. In respect to demographic characteristics, there were no significant differences between patients who achieved four weeks of abstinence from cocaine and those who did not. Baseline variables that most consistently predicted cocaine abstinence included initial urine drug screen (UDS) results, the initial Cocaine Selective Severity Assessment (CSSA) scores, and initial self-reported cocaine use in past 30 days, whereas cocaine craving, cocaine composite scores, alcohol craving, alcohol withdrawal symptoms, and alcohol composite scores did not. The results of this study suggest that cocaine dependence severity in general, and initial UDS results, the CSSA scores and frequency of recent cocaine use in particular, have a significant impact on treatment outcome in the treatment of cocaine-dependent patients with comorbid alcoholism. Initial UDS results and CSSA scores are very useful predictors of treatment outcome and could be used as stratifying variables in outpatient cocaine and alcohol medication trials. PMID- 19219672 TI - Regular exercise as a protective factor in relapse following smoking cessation treatment. PMID- 19219673 TI - The introduction to Japan of the Titan barnacle, Megabalanus coccopoma (Darwin, 1854) (Cirripedia: Balanomorpha) and the role of shipping in its translocation. AB - The Titan Acorn barnacle, Megabalanus coccopoma, a native of the tropical eastern Pacific, has become established in the western Atlantic (Brazil and the northern Gulf of Mexico to the Carolinas), northwestern Europe and the western Indian Ocean (Mauritius), and therefore its dispersal capabilities are well known. This study reports its introduction to Japan and confirms its occurrence in Australia. In an attempt to determine the source of this introduction, phylogeographic techniques, involving cytochrome c oxidase I sequences of various widely separate populations of M. rosa and M. volcano, were utilized. No significant genetic differentiation or haplotype patterns between widely separated populations of each of the three species were found. Lack of such differentiation indicates recent geographical isolation and thus negates a null hypothesis predicting that the occurrence of one of more of these species in Australia was natural. PMID- 19219674 TI - Phase I study of weekly cisplatin, bolus fluorouracil and escalating doses of irinotecan in advanced solid tumors. AB - PURPOSE: We performed a phase I study of 5-fluorouracil (5-FU), cisplatin and irinotecan. METHODS: Twenty-nine patients received cisplatin 25 mg/m(2) and bolus 5-FU 425 mg/m(2), along with irinotecan at 40, 50, and 65 mg/m(2) weekly for 4 out of 6 weeks. RESULTS: The maximum tolerated dose (MTD) for untreated patients was irinotecan 65 mg/m(2) while the MTD for previously treated patients was irinotecan 40 mg/m(2). Neutropenia and diarrhea were the major dose-limiting toxicities. Antitumor activity was noted in gastric, esophageal and pancreatic cancers. CONCLUSION: Because of the toxicity profile, combinations with continuous infusion 5-FU or capecitabine should be explored. PMID- 19219676 TI - Expression profile of human beta-defensin 3 in oral squamous cell carcinoma. AB - Although it is known that innate immunity is important for protecting the body against foreign agents such as bacteria, little is known about elements of the innate immune system that have antitumor activity. This prospective study was designed to investigate the function of human beta-defensin 3 (hBD-3), an important component of the innate immune response, in oral squamous cell carcinoma (OSCC). Paired cancerous and noncancerous specimens of 45 patients who underwent surgical treatment for OSCC were examined for hBD-3 expression on protein and mRNA. Clinical and pathological features such as age, gender, tumor and lymph node status, UICC stage, and histological grading were correlated. hBD 3 was significantly overexpressed in tumors in comparison to healthy tissue examined with real-time quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) analysis (p = .004). Immunohistochemical stain for hBD-3 was much more pronounced in tumors than in corresponding healthy mucosa. The results illustrate that hBD-3 is frequently overexpressed in oral squamous cell carcinomas and seems to be related to oncogenesis. Increased expression of hBD-3 in oral squamous cell carcinomas suggests its potential role in the pathogenesis of oral cancer. This might be a starting point for novel pharmacological/molecular treatment modalities. PMID- 19219675 TI - A first report of radioembolization for hepatic metastases from ocular melanoma. AB - BACKGROUND: Ocular melanoma (OM) metastasizes to the liver and is rapidly fatal despite aggressive therapy. Yttrium-90 microspheres (radioembolization) delivered via the hepatic artery is an established and effective approach for primary and metastatic hepatic tumors, although (90)Y use in OM has not been reported previously. METHODS: A retrospective review was performed for all patients with OM who received radioembolization at 5 centers. RESULTS: 11 patients received 12 treatments with a median activity of 1.55 GBq delivered per treatment. Toxicity was minimal, with PET/CT at 3 months posttreatment showing a response in all patients; 1 patient had a complete response. CONCLUSIONS: Radioembolization can control hepatic metastases of OM with very few side effects. PMID- 19219677 TI - Stresses in cement mantles of hip replacements: effect of femoral implant sizes, body mass index and bone quality. AB - The effects of femoral prosthetic heads of diameters 22 and 28 mm were investigated on the stability of reconstructed hemi-pelves with cement mantles of thicknesses 1-4 mm and different bone qualities. Materialise medical imaging package and I-Deas finite element (FE) software were used to create accurate geometry of a hemi-pelvis from CT-scan images. Our FE results show an increase in cement mantle stresses associated with the larger femoral head. When a 22 mm femoral head is used on acetabulae of diameters 56 mm and above, the probability of survivorship can be increased by creating a cement mantle of at least 1 mm thick. However, when a 28 mm femoral head is used, a cement mantle thickness of at least 4 mm is needed. Poor bone quality resulted in an average 45% increase in the tensile stresses of the cement mantles, indicating resulting poor survivorship rate. PMID- 19219678 TI - The anti-inflammatory effect of subconjunctival bevacizumab on chemically burned rat corneas. AB - PURPOSE: To investigate the effect of bevacizumab in a model of corneal inflammation. MATERIALS AND METHODS: Epithelium from the cornea and limbus was completely removed using 100% alcohol in rats. Bevacizumab (1.25 mg/0.05 ml) or normal saline was subconjunctivally injected. One week later, corneas were examined and subjected to hematoxylin-eosin and immunofluorescent staining for VEGF. Expression of IL-2, IFN-gamma, IL-6, and TGF-beta 1 were analyzed by ELISA. RESULTS: Bevacizumab showed a borderline reduction in corneal neovascularization (11.0 +/- 4.8% and 18.0 +/- 10.0% in the bevacizumab and control groups, respectively; p = 0.054), while the extent of epithelialization was not affected (5.0 +/- 3.4% and 6.1 +/- 5.2% in the bevacizumab and control groups, respectively; p = 0.715). The infiltration of inflammatory cells was reduced (99.3 +/- 22.3 cells/x 400 in the bevacizumab-injected corneas and 182.3 +/- 58.0 cells/x 400 in the controls; p = 0.013). The levels of IL-2, IFN-gamma, and IL-6 were decreased in the rats with the bevacizumab injections (44 +/- 6 and 79 +/- 9 pg/ml for IL-2 in the bevacizumab-injected group and control, respectively, p = 0.025; 45 +/- 5 and 67 +/- 6 pg/ml for IFN-gamma in the bevacizumab-injected group and controls, respectively, p = 0.043; 45 +/- 6 and 75 +/- 8 pg/ml for IL-6 in the bevacizumab-injected group and controls, respectively, p = 0.030). CONCLUSIONS: Bevacizumab showed a reduction in inflammatory cell infiltration and cytokines in chemically burned rat corneas. PMID- 19219679 TI - Protein array characterization of bioactive proteins secreted by immortalized human corneal epithelium in response to pseudomonas constituents. AB - PURPOSE: To use protein arrays to delineate the spectrum of angiogenic bioactive protein modulators that might be secreted and up-regulated by the corneal epithelium in response to killed bacterial products. METHODS: Immortalized human corneal epithelial cells were grown in culture, serum starved, and exposed to heat-killed Pseudomonas aeruginosa in a dose-dependent manner. The resultant culture medium was screened by antibody arrays for 43 proteins that can modulate angiogenesis and immune and inflammatory processes. Parallel analysis was carried out on tears recovered in the open and closed eye phases (OTF and CTF) of the diurnal cycle. RESULTS: Array analysis reveals that the immortalized cells constitutively secrete several proteins and up-regulate the secretion of IL-6, IL 8, and GRO in response to killed bacteria. Also evident was the emergence of a strong signal for GM-CSF and moderate/weak signals for MCP-1, MMP-9, Leptin, and INF gamma in a dose-dependent manner. Several of these proteins, including IL-6, IL-8, GRO, MMP-9, TIMP-1, and MCP-1, accumulate in the CTF. Other proteins are unique to tear fluid. CONCLUSIONS: Nine proteins were identified that are secreted by epithelium in response to killed bacteria that contribute to the innate and adaptive defense system through potentiating PMN and macrophage recruitment, activation, and opsonization in a cooperative manner. The vast majority of these proteins are angiogenic modulators, perhaps contributing to the imbalance between angiogenic and angiostatic processes and risk of corneal vascularization. PMID- 19219680 TI - A human apolipoprotein E mimetic peptide effectively inhibits HSV-1 TK-positive and TK-negative acute epithelial keratitis in rabbits. AB - PURPOSE: To compare the antiviral effect of a peptide derived from human apolipoprotein E (1% apoEdp) with 1% trifluorothymidine against HSV-1 thymidine kinase (TK)-positive (HSV-1 McKrae) and with 3% foscarnet against HSV-1 TK negative (KOS background) in the rabbit eye model of acute HSV-1 epithelial keratitis. MATERIALS AND METHODS: Topical treatment began three days post infection and continued for five days. Rabbits were treated 5 times per day with 1% apoEdp, 1% trifluorothymidine, 3% foscarnet, or PBS. Slit lamp examination was performed on a masked basis from post-infection days 3-10 to determine the severity of epithelial keratitis. RESULTS: Topical treatment with 1% apoEdp was as effective as 1% trifluorothymidine against TK-positive and 3% foscarnet against TK-negative HSV-1 epithelial keratitis. CONCLUSIONS: ApoEdp 1% has efficacy for the topical treatment of TK-positive and TK-negative ocular herpes in the rabbit eye model. Thus, apoEdp has the potential to be used for treatment of keratitis caused by TK mutants. PMID- 19219681 TI - ERK1/2 activation regulates the wound healing process of rabbit corneal endothelial cells. AB - PURPOSE: To evaluate the activation of ERK, a MAP kinase, in the regulation of proliferation and migration of rabbit corneal endothelial cells during the healing process after scrape wound injury. METHODS: Cultured rabbit corneal endothelial cells were used in this study. Immunocytochemistry and Western blot analysis were performed to demonstrate the phosphorylation of ERK after scrape injury. The injured cell sheets were treated with different concentrations of ERK inhibitor, PD98059. The wound closure at 24, 48, and 72 hr following injury was evaluated. In order to evaluate the role of proliferation during wound healing, immunocytochemistry with Ki67 and 3-(4,5-dimethylthiazol-2-yl)-5-(3 carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt (MTS) assay were performed. The role of cell migration in wound healing was evaluated by quantification with the time lapse microscopic video imaging system. RESULTS: Immunocytochemical and Western blot analysis revealed phosphorylation of ERK in the rabbit corneal endothelial cells after wounding. Inhibition of ERK phosphorylation by PD58059 resulted in a dose-dependent delay of wound closure. The inhibition of wound healing became significant from 48 to 72 hr in cells treated with 10 and 50 micro M of PD58059 (p < 0.05). ERK inhibition suppressed cell proliferation as indicated by Ki67 immunostaining and MTS assay. The time lapse microscopic video imaging analysis indicated delayed cell migration caused by PD98059. CONCLUSIONS: ERK activation regulates wound repair of rabbit corneal endothelial cells. Both cell proliferation and migration are involved in this process. PMID- 19219682 TI - Expression of fibroblast activation proteins in corneal stromal neovascularization. AB - PURPOSE: To observe changes of the factors in corneal stroma during corneal neovascularization and to investigate the mechanism of corneal neovascularization. METHODS: Forty-eight Wistar rats were used, including eight in the control group. Corneal neovascularization was induced by alkali burns in 40 rats. Frozen sections, which were cut across the corneal center, were prepared on days 1, 3, and 7 post-burn, respectively. Transforming growth factor-beta 1 (TGF-beta 1) was examined by immunohistochemistry, and fibroblast activation protein (FAP) and a-smooth muscle actin (a-SMA) were detected by double-labeling fluorescent immunohistochemistry. Blood vessel endothelium was identified for PECAM-1 (CD31). Expressions of FAP in the cornea with or without neovascularization were monitored with reverse transcription-polymerase chain reaction on days 3 and 7. RESULTS: In the alkali-burned eyes, TGF-ss1 first expressed in the corneal stroma, and some stromal cells expressed a-SMA and FAP. The FAP(+) keratocytes were found around the CD31(+) endothelium of angiogenesis. FAP was expressed in the corneas with neovascularization, but not in those without neovascularization. CONCLUSION: Factors in corneal stroma may change when corneal neovascularization occurs. The stromal keratocytes can express FAP(+) cells surrounding the endothelium of angiogenesis. PMID- 19219683 TI - Comparative study of plasma ascorbic acid levels in senile cataract patients and in normal individuals. AB - OBJECTIVE: Dietary antioxidant vitamins, in particular vitamin C, could play a role in preventing the onset or progression of age-related visual impairment. The aim of this study was to determine the plasma ascorbic acid levels in normal individuals and in senile cataract patients. METHOD: The test group included 50 cataract patients, while the control group members were selected among medical staff and patients' companions after age matching with the test group. Questionnaire forms contained age, sex, and location of residency (urban or rural) inquiries. Five milliliters of drawn blood was centrifuged, and plasma ascorbic acid level was measured by spectrophotometer. Statistical analysis of plasma ascorbic acid concentrations were done by paired test. RESULTS: The mean plasma ascorbic acid level in the test group was 0.96 +/- 0.08 mg/dl and 1.12 +/- 0.15 mg/dl in the control group. Ascorbic acid level in the test group was significantly lower than the control group and p value = 0.00001. Aging had a reverse relation with plasma ascorbic acid level in males, while it showed no significant alterations in females. Rural compared with urban and females compared to males showed higher levels of plasma ascorbic acid. CONCLUSION: This study revealed that plasma ascorbic acid level in cataract patients was lower than normal individuals. Antioxidant vitamins, in particular vitamin C, found in Mediterranean fruits and vegetables, can help with the prevention of cataracts, which is a major health service burden in many countries. PMID- 19219684 TI - Expression of macrophage colony-stimulating factor (M-CSF) and its receptor in streptozotocin-induced diabetic rats. AB - PURPOSE: To demonstrate the expression and location of macrophage colony stimulating factor (M-CSF) and its receptor (CSF-1R) in the retinas of diabetic rats, as well as in vitreous human proliferative diabetic retinopathy (PDR). METHODS: The retinas of streptozotocin-induced diabetic rat were studied. Real time PCR was applied to evaluate M-CSF and its receptor CSF-1R mRNA expression in the retinas. The protein levels of M-CSF and CSF-1R were evaluated by Western blot analysis. Cellular sources of M-CSF and CSF-1R were determined by double immunofluorescence staining. M-CSF levels in vitreous samples from patients with PDR were measured by ELISA. RESULTS: M-CSF and CSF-1R mRNA were upregulated in the retinas as early as two weeks after the onset of diabetes and increased over time. A similar pattern was observed for M-CSF/CSF-1R protein expression levels. Double-immunofluorescence staining revealed that increased M-CSF immunoreactivity occurred mainly in the nerve fiber layer in diabetic retinas, co-localizing with glial fibrillary acidic protein. Increased CSF-1R immunoreactivity was observed in OX-42-labeled microglia and ganglion cells in the ganglion cell layer. The vitreous level of M-CSF was elevated in patients with PDR compared to control subjects. CONCLUSIONS: The early upregulation of MCSF/CSF-1R signaling may be an important regulatory pathway among neurons, microglia, and glia in diabetic retinopathy. PMID- 19219685 TI - Extracellular matrix proteins in epiretinal membranes and in diabetic retinopathy. AB - PURPOSE: Non-vascular epiretinal membranes (ERM) and neovascular membrane in proliferative diabetic retinopathy (PDR) are recognized causes of visual impairment. Both ERMs and neovascular membranes in PDR consist of cellular components and extracellular matrix (ECM) proteins such as fibronectin (FN) and collagen. Transforming growth factor-beta (TGF-beta) and endothelin-1 (ET-1) regulate ECM protein production. In this study, we investigated ECM proteins and their regulators in ERMs and vitreous from PDR subjects and non-diabetic subjects undergoing vitrectomy. METHODS: ERMs from non-diabetic subjects undergoing membrane peeling were collected. Vitreous samples from non-diabetic and PDR subjects undergoing vitrectomy were also collected and separated into solid pellets consisting of fibrovascular tissue and vitreous fluid. Real-time PCR was done for estimating mRNA levels of extracellular matrix proteins like collagen, FN, its splice variant extra-domain B containing FN (EDBFN), and their regulators, TGF-beta and ET-1. ELISA was done to detect the EDBFN level in blood and vitreous from non-diabetic and PDR subjects undergoing vitrectomy. RESULTS: ECM proteins, including FN, its splice variant EDBFN, and collagen were significantly upregulated in the ERMs and PDR compared to vitreous from both other two group. The levels were, however, higher in the ERM. ECM protein regulators like TGF-beta and ET-1 were also elevated. FN and EDBFN show significant correlation with TGF-beta in vitreous but not in ERMs. Plasma and vitreous EDBFN were elevated in the PDR subjects compared to non-diabetic subjects. CONCLUSIONS: Data from these studies show that ECM proteins such as EDBFN and collagen are upregulated in ERM and PDR, and are regulated by TGF-beta. Elevated serum EDBFN in the PDR may potentially be further explored as a possible molecular marker for the early detection of diabetic end organ damages. PMID- 19219686 TI - Effect of intravitreal triamcinolone acetonide in the treatment of experimental Staphylococcus epidermidis endophthalmitis. AB - PURPOSE: To investigate the effect of intravitreal injection of triamcinolone acetonide (TA) in the treatment of experimental Staphylococcus epidermidis endophthalmitis. MATERIALS AND METHODS: Experimental endophthalmitis induced in one eye of 38 rabbits by intravitreal injection of Staphylococcus epidermidis. The infected rabbits were randomly divided into three groups. The rabbits in group 1 (n = 15) received intravitreal injection of 1 mg vancomycin hydrochloride. Group 2 eyes (n = 15) received intravitreal TA (4 mg) plus vancomycin hydrochloride (1 mg). Group 3 eyes (8 eyes) received normal saline. Clinical examination in each group was performed two, seven, and 15 days after inoculation of the bacteria. The eyes were enucleated for histopathologic evaluation on the 15th day. Clinical and histological scoring systems were used to evaluate clinical and histological severity of the intraocular infection. RESULTS: The mean clinical scores of three study groups at the 7th day were 5.8 +/- 2.3, 3.3 +/- 1.5, and 9.5 +/- 2.1, respectively. The mean of this value at 15th day were 6.2 +/- 2.6, 3.9 +/- 2.1, and 10 +/- 2.2, respectively. At the 7th and 15th day, the mean clinical score of group 2 was significantly lower when compared with the other two groups. Mean histopathologic scores of group 1, group 2, and group 3 were 11 +/- 2.7, 7.9 +/- 2.9, and 14.0 +/- 3.1, respectively. Histopathologic studies revealed less tissue destruction for eyes treated with TA. There was a significant positive correlation between the severity of clinical signs and pathologic damage in all three experiment groups; however, it was weakest in the IVTA treated group. CONCLUSION: A standard clinical dose of intravitreal TA attenuates the clinical signs and tissue destruction in a rabbit model of Staphylococcus epidermidis endophthalmitis. PMID- 19219687 TI - Vitreous and serum levels of platelet-derived growth factor and their correlation in patients with proliferative diabetic retinopathy. AB - PURPOSE: We investigated possible correlations between vitreous and/or serum levels of platelet derived growth factor isoforms (PDGF-AA, -AB, -BB) with parameters associated with proliferative diabetic retinopathy (PDR), and compared the results to vascular endothelial growth factor (VEGF), which is an established growth factor affecting PDR. METHODS: Thirty-one patients with PDR and 15 non diabetic patients were included in the study. Vitreous and serum samples were obtained during vitrectomy. PDGF-AA, -AB, and -BB, as well as VEGF, were measured by enzyme-linked immunosorbent assay. RESULTS: PDGF-AA, -AB, -BB, and VEGF were all expressed in serum and vitreous of controls and patients with PDR. The levels of all PDGF isoforms and VEGF in vitreous were significantly increased in the PDR group, as compared to controls. No such differences were evident in serum. PDGF AA and PDGF-BB correlated significantly to the severity but not the activity of PDR. PDGF-AB and -BB were significantly lower in vitreous of patients with pre performed complete panretinal photocoagulation (PRP) as compared to incomplete or without PRP. PDGF did not correlate significantly to fibrovascular tissue, on the disc or elsewhere, to long-standing vitreous hemorrhage, to tractional retinal detachment, or to posterior vitreous detachment. PDGF or VEGF in vitreous or serum of PDR patients did not correlate with the serum levels of HbA1C. There was no correlation between the vitreous and serum levels of VEGF or PDGF in patients with PDR. CONCLUSIONS: It appears that, in addition to VEGF, PDGF-AA, -AB, and BB in the vitreous are also correlated with PDR. PMID- 19219688 TI - Reproducible measurement of macular light flash recovery time using a novel device can indicate the presence and worsening of macular diseases. AB - PURPOSE: To determine the safety, sensitivity, and specificity of a novel flash photorecovery timing instrument with response verification in differentiating normal from abnormal maculae, and in detecting worsening macular disease. METHODS: Right and left eye photorecovery times were determined at baseline and after 5 min using a xenon arc, flash filtered for infrared, ultraviolet, and visible short wavelengths, delivered through an aperture in a hand-held tube. A push-button actuated timer and flash and stopped timer when lighted numbers became visible post-flash. A numeric keypad verified responses. Normal subjects (two eyes tested, n = 144; one eye tested, n = 108) ranged in age from 15 to 84. Photorecovery times were measured in one eye of subjects with small drusen and 20/20 acuity (53-55 correct ETDRS letters; n = 57); in both eyes of subjects with dry age-related macular degeneration (AMD; n = 118); wet AMD with (n = 19) or without (n = 17) macular fluid; and eyes of diabetics with background retinopathy with (n = 19) or without (n = 17) macular retinal thickening. Once-weekly photorecovery measurements for 6 months in each eye of 10 dry AMD subjects and 10 dry diabetic maculopathy subjects provided longitudinal data. RESULTS: Normal subjects' mean right eye recovery time was 9.6 sec (+/- 1.9 SD); left 10.8 sec (+/- 1.0 SD). Photorecovery lengthened after age 55, nearly doubling that of young subjects by age 80. Macular edema, serous macular detachment, or worsened dry AMD were accompanied by prolonged photorecovery (p < .01). When abnormal new vessels or retinal thickening appeared in three serially followed patients, photorecovery at least doubled (p < .01). In all three, photorecovery prolongation occurred without clinical symptoms. None of the 499 tested subjects reported adverse events due to the flash testing. CONCLUSIONS: These findings support the usefulness of a reproducible light flash macular vision recovery measurement as an indicator of macular pathology and worsening disease. PMID- 19219689 TI - Effect of presentation level on diagnosis of dead regions using the threshold equalizing noise test. AB - The effect of the level of threshold equalizing noise (TEN) on the diagnosis of dead regions (DRs) was investigated. Participants comprised 23 adults with sensorineural hearing impairment. Masked thresholds were measured monaurally with TEN at 60, 70, 80, and 90 dB HL/ERB(N). Absolute and masked thresholds (with TEN at 80 dB HL/ERB(N)) were retested. The diagnosis was unaffected by TEN level at any frequency for eight of the 13 participants who met the criteria for a DR. For four of the latter, increasing the TEN level changed the diagnosis from DR to no DR, mainly at 1.5 kHz, corresponding to the edge frequency, f(e), of the DR. For one participant with a low-frequency DR, increasing the TEN level changed the diagnosis from no DR to DR, only at 1 kHz. The changes with level were too large to be explained in terms of test repeatability. Overall, the results of the TEN test are usually not affected by TEN level, except for test frequencies close to f(e) when absolute thresholds are near-normal for frequencies adjacent to f(e). PMID- 19219690 TI - Comparison of two frequency-to-electrode maps for acoustic-electric stimulation. AB - The fitting of a cochlear implant together with aided residual hearing was evaluated by means of matching frequency and/or perceived pitch between acoustic and electric modalities. Five cochlear implant users with the Nucleus Freedom electrode array with residual acoustic hearing participated. Psychophysical procedures were used to create a map in which the implant was programmed to provide the listener with high-frequency information only above the frequency at which acoustic hearing was no longer considered useful. This was compared to a second map which provided the full frequency range. Listeners wore each map for a number of weeks before speech recognition was measured in quiet and noise. Post operatively across subjects, average hearing thresholds worsened by 27 dB. However, cochlear implantation provided superior recognition of speech compared to pre-operative scores, with the best results found when subjects were wearing their hearing aids together with the implant. No significant differences were found between the two maps on speech tests when subjects were wearing their implant together with hearing aid/s. In conclusion, the combination of a cochlear implant together with hearing aid/s was effective at providing speech perception benefits for the listeners of the current study, regardless of the frequency-to electrode allocation selected. PMID- 19219691 TI - Access to cochlear implant candidacy evaluations: who is not making it to the team evaluations? AB - The objective of this study was to investigate trends in the referral process among pediatric cochlear implant candidates. Medical and audiologic charts between 2003 and 2005 were reviewed, and children five years and younger with moderately-severe or worse sensorineural hearing loss were included. Of the 105 audiograms meeting the inclusion criteria, 69% were referred for a cochlear implant, and 52% were considered as definite candidates for an implant by audiologists with expertise in cochlear implant technology. Children referred for an implant, compared to children who were not referred, were more likely to have married parents (91% vs. 70%, p=0.02) and more likely to have private insurance (56% vs. 29%, p=0.02). Multivariable regression results were consistent with the unadjusted findings regarding marital status, but not insurance status. Children with sensorineural hearing loss are inconsistently referred to cochlear implant teams despite similar audiologic findings. To reach the Healthy People 2010 goals, this disparity should be addressed. A further understanding of the population of children not referred is important in diminishing inconsistencies and understanding barriers to care. PMID- 19219692 TI - Automatic testing of speech recognition. AB - Speech reception tests are commonly administered by manually scoring the oral response of the subject. This requires a test supervisor to be continuously present. To avoid this, a subject can type the response, after which it can be scored automatically. However, spelling errors may then be counted as recognition errors, influencing the test results. We demonstrate an autocorrection approach based on two scoring algorithms to cope with spelling errors. The first algorithm deals with sentences and is based on word scores. The second algorithm deals with single words and is based on phoneme scores. Both algorithms were evaluated with a corpus of typed answers based on three different Dutch speech materials. The percentage of differences between automatic and manual scoring was determined, in addition to the mean difference in speech recognition threshold. The sentence correction algorithm performed at a higher accuracy than commonly obtained with these speech materials. The word correction algorithm performed better than the human operator. Both algorithms can be used in practice and allow speech reception tests with open set speech materials over the internet. PMID- 19219693 TI - Pediatric cochlear implantation: how much hearing is too much? AB - Audiologic candidacy criteria for determining cochlear implantation candidacy in children are evolving. The objective of the study was to examine clinical practice related to the cochlear implantation of children who typically do not meet audiologic criteria for this technology. Practitioners' perspectives on the process and the factors influencing candidacy decisions were explored through focus group interviews with hospital and school-based practitioners. The interviews were analysed using qualitative techniques to identify key issues. The findings from the interviews informed a questionnaire which was sent to all cochlear implant centers in Canada to further examine clinician views and experiences with this special population. Responses were collected from 11 of the 12 centers and indicated that children with hearing outside typical criteria were receiving implants. The definition of 'borderline' varied across the programs from approximately 70 dB HL to less than 90 dB HL. All centers emphasized the importance of considering factors beyond the child's audiometric thresholds in candidacy decision-making. PMID- 19219695 TI - Arrhenius relationships from the molecule and cell to the clinic. AB - There are great differences in heat sensitivity between different cell types and tissues. However, for an isoeffect induced in a specific cell type or tissue by heating for different durations at different temperatures varying from 43-44 degrees C up to about 57 degrees C, the duration of heating must be increased by a factor of about 2 (R value) when the temperature is decreased by 1 degrees C. This same time-temperature relationship has been observed for heat inactivation of proteins, and changing only one amino acid out of 253 can shift the temperature for a given amount of protein denaturation from 46 degrees C to either 43 or 49 degrees C. For cytotoxic temperatures <43-44 degrees C, R for mammalian cells and tissues is about 4-6. Many factors change the absolute heat sensitivity of mammalian cells by about 1 degrees C, but these factors have little effect on Rs, although the transition in R at 43-44 degrees C may be eliminated or shifted by about 1 degrees C. R for heat radiosensitization are similar to those above for heat cytotoxicity, but Rs for heat chemosensitization are much smaller (usually about 1.1-1.2). In practically all of the clinical trials that have been conducted, heat and radiation have been separated by 30-60 min, for which the primary effect should be heat cytotoxicity and not heat radiosensitization. Data are presented showing the clinical application of the thermal isoeffect dose (TID) concept in which different heating protocols for different times at different temperatures are converted into equivalent minutes (equiv) min at 43 degrees C (EM(43)). For several heat treatments in the clinic, the TIDs for each treatment can be added to give a cumulative equiv min at 43 degrees C, namely, CEM(43). This TID concept was applied by Oleson et al. in a retrospective analysis of clinical data, with the intent of using this approach prospectively to guide future clinical studies. Considerations of laboratory data and the large variations in temperature distributions observed in human tumors indicate that thermal tolerance, which has been observed for mammalian cells for both heat killing and heat radiosensitization, probably is not very important in the clinic. However, if thermal tolerance did occur in the clinical trials in which fractionation schemes were varied, it probably would not have been detected because with only the two-three-fold change in treatment time that occurs when comparing one versus two fractions per week, or three versus six total fractions, little difference would be expected in the response of the tumors since both thermal doses were extremely low on the dose-response curve. Data are shown which indicate that in order to test for thermal tolerance in the clinic and to have a successful phase III trial, the thermal dose should be increased about five-fold compared with what has been achieved in previous clinical trials. This increase in thermal dose could be achieved by increasing the temperature about 1.5 degrees C (from 39.5 to 41.0 degrees C in 90% of the tumor) or by increasing the total treatment time about five-fold. The estimate is that 90% of the tumor should receive a cumulative thermal dose (CEM(43)) of at least 25; this is abbreviated as a CEM(43) T(90) of 25. This value of 25 compares with 5 observed by Oleson et al. in their soft tissue sarcoma study. Arguments also are presented that thermal doses much higher than the CEM(43) T(90) induce the hyperthermic damage that causes the tumors to respond, and that the minimum CEM(43) T(90) of 25 only predicts which tumors that receive a certain minimal thermal dose in <90% of the regions of the tumors will respond. For example, in addition to a minimal CEM(43) T(90) of 25 a minimum CEM(43) T(50) of about 400 also may be required for a response. Finally, continuous heating for approximately 2 days at about 41 degrees C during either interstitial low dose-rate irradiation or fractionated high dose-rate irradiation, which we estimate could give a CEM(43) of 75, should be considered in order to enhance heat radiosensitization of the tumor as well as heat cytotoxicity. In order to exploit the use of hyperthermia in the clinic, we need a better understanding of the biology and physiology of heat effects in tumors and various normal tissues. As an example of an approach for mechanistic studies, one specific study is described which demonstrates that damage to the centrosome of CHO cells heated during G(1) causes irregular divisions that result in multinucleated cells that do not continue dividing to form colonies. This may or may not be relevant for heat damage in vivo. However, since normal tissues vary in thermal sensitivity by a factor of 10, similar approaches are needed to describe the fundamental lethal events that occur in the cells comprising the different tissues. PMID- 19219696 TI - Hyperthermia classic commentary: 'Arrhenius relationships from the molecule and cell to the clinic' by William Dewey, Int. J. Hyperthermia, 10:457-483, 1994. PMID- 19219697 TI - Immunologic response induced by synergistic effect of alternating cooling and heating of breast cancer. AB - In this study, the therapeutic effect and the induced anti-tumor immunity through the alternating cooling and heating was investigated using 4T1 murine mammary carcinoma, a common model of human metastatic breast cancer. While fifteen of seventeen regular mice were cured, primary recurrence and metastasis caused death of all the nude mice within one month after the same treatment. Histological analyses showed that viable cells existed in the tumor debris after the treatment, indicating that the direct killing effect was not the only therapeutic mechanism. Further investigation found rejection of tumor upon re-challenge, and anti-tumor immune response was studied. Stronger cytotoxicity T-lymphocyte (CTL) and Th1 cytokines response as well as infiltration of immunocytes were observed in the treated mice in comparison to those after the surgical resection. The results showed that the alternating cooling and heat could stimulate anti-tumor immunologic response in vivo and the underlying mechanisms will be further investigated in the near future. PMID- 19219698 TI - Heat reactions in multiple sclerosis: an overlooked paradigm in the study of comparative fatigue. AB - Multiple sclerosis (MS) is a demyelinating and debilitating disease characterised by a range of symptoms such as motor dysfunction and muscle weakness. A significant MS symptom is heat sensitivity so that exposure to heat will increase body temperature and consequently the appearance of neurological signs. Although some people with MS can undertake exercise, it is thought to be limited by the sensitivity to heat and the subsequent rise in body temperature which occurs. It has been found that central fatigue is a determining factor in muscle activation and performance in normal healthy subjects. However, it is unknown whether thermal strain also induces central fatigue in MS even though muscular fatigue in MS is due mainly to central rather than peripheral factors. This review focuses on the similarities in the manifestation of central fatigue in both MS and healthy subjects with reference to thermal strain and heat reactions. PMID- 19219699 TI - Kinetics study on markers of the immune system by gene expression profiling of an in vivo heated tumor. AB - PURPOSE: To analyze effects of hyperthermia on immune cells within tumors. MATERIALS AND METHODS: Subcutaneously implanted rat gliomas were treated with water-bath hyperthermia for one hour at day zero (HT) (intratumoral temperature of 43 degrees C), low-dose metronomic cyclophosphamide (CTX) 35 mg/kg three times a week for two weeks, both HT and CTX (CTX-HT(0)), or saline. Tumors harvested at day 1, 7, 14 and 21 were analyzed for changes in gene expression by microarrays, focusing on genes expressed in immune cells. Microarray analyses and real-time RT PCR were previously performed on tumors harvested day zero at 0, 45, 90 and 180 minutes after end of treatment. Gene expression kinetics of selected genes were analyzed in all tumors by quantitative real-time RT-PCR (qPCR) to validate the results of the microarrays. RESULTS: Previous microarray analyses have indicated a suppression of gene expression in immune cells within tumors by hyperthermia the first three hours after treatment. qPCR analyses of CD14, CD36, Klrd1 (CD94), Tlr2 and Lrp1 confirmed these results. Global mRNA screen revealed no hyperthermia-induced changes in gene expression of immune cells at day 1, 7, 14 and 21. qPCR analyses of CD14, CD36, Klrd1 (CD94), Tlr2 and Lrp1 confirmed the results of the microarrays, i.e. none of these were differentially expressed after the first day, with the exception of Lrp1 which displayed elevated mRNA levels. CONCLUSION: The suppression in gene expression of a range of immune cells within tumors treated with hyperthermia at 43 degrees C is a transient phenomenon. PMID- 19219700 TI - Thermal ablation of the goat mammary gland as a model for post-lumpectomy treatment of breast cancer: preliminary observations. AB - BACKGROUND: Partial breast irradiation post-lumpectomy, with a balloon bearing a radioactive source in its center, is practiced as an alternative to whole breast irradiation in the treatment of breast cancer. The goal is to ablate residual malignant cells within 1 cm radius of the resected lumpectomy margin. We hypothesize that this goal may be achieved with a fluid-filled heated balloon. METHODS: Nubian-cross goats were treated under general anesthesia. The two mammary glands were sequentially bisected and a non-inflated balloon with a heating element was placed in the center of the gland which was re-sutured. Two series of experiments were conducted. In the first 22 goats (44 glands), the balloon was inflated with 5% dextrose to a pressure of 150 mmHg and heated at 87 degrees C over selected time intervals of 1-24 minutes. In the second series (16 glands), the re-programmed device operated at 50-80 mmHg over selected time intervals of 5-20 minutes. The depth of necrosis was histologically determined after sacrificing the goats and excising the glands. RESULTS: In the first series, glandular necrosis was noted to extend to a depth of 3.2-9.6 mm for the above heating cycles. Corresponding figures for the second series ranged from 4.7 8.6 mm for treatment times of one minute 'warm up' to 20 minutes of heating at 90 degrees C. The animals exhibited no systemic side effects post-treatment. CONCLUSION: An experimental model describing a thermal technique causing necrosis of the goat mammary gland is described. PMID- 19219701 TI - Measuring urethral tissue heat injury temperature of healthy male rabbits during interstitial radiofrequency ablation. AB - OBJECTIVE: The objective was to evaluate urethral tissue heat injury in a rabbit model. Histopathology analysis was used to determine the minimum temperature required to achieve urethral tissue injury following interstitial radiofrequency ablation (RFA). MATERIALS AND METHODS: A total of 37 healthy rabbits were divided into 6 groups randomly and treated with interstitial RFA in the penial parenchyma. The temperatures of urethra were monitored and controlled to 48 degrees , 49 degrees , 50 degrees , 51 degrees 52 degrees and 53 degrees C respectively. The urethral tissue acute heat injury (48 h after heating) was assessed by HE stain, Terminal Deoxylnucleotidyl Transferase Mediated-dUTP Nick End Labeling (TUNEL) stain and quantitative analysis in tissue sections. RESULTS: Histologically, the main feature of acute heat damage was necrosis or vascular congestion or thrombosis of blood vessels of the urethral wall. This occurred only in one out of five cases at 49 degrees C and 50 degrees C heating, but in four out of five cases at 52 degrees and 53 degrees C. The percentage of necrosis was significantly different at tissue temperature of 52 degrees C and above. Quantitative image analyses of TUNEL stain sections demonstrated a significant increase in the positive staining for apoptotic cell at tissue temperatures of 50 degrees C and above. It indicated that the TUNEL stain to detect cell death was more sensitive than routine histology. CONCLUSION: The results from this in vivo study indicate that 50 degrees C and 5 min heating of rabbit urethra during interstitial RFA is the minimum temperature for heat injury of the normal rabbit urethra as measured at 48 h after treatment. PMID- 19219702 TI - Effects of hyperthermia with dextran magnetic fluid on the growth of grafted H22 tumor in mice. AB - OBJECTIVE: To study the effect of dextran magnetic fluid hyperthermia on growth, vascular endothelial growth factor (VEGF) expression and apoptosis of the grafted H22 tumors in mice. METHODS: Mice with grafted H22 tumor were randomly divided into four groups. The treatment group was intratumorally injected with dextran magnetic fluid and subsequently exposed to a 55 kHz 20 kA/m alternating magnetic field for 10 min. The three control groups were respectively injected with physiological saline and no magnetic field; with physiological saline and exposed to 55 kHz 20 kA/m alternating magnetic field for 10 min; and with dextran magnetic fluid only. The inhibitive ratios were calculated. Other mice with grafted tumor were randomly divided into two groups, respectively injected with dextran magnetic fluid or physiological saline and exposed to a 55 kHz 20 kA/m alternating magnetic field for 10 min. The VEGF expression and apoptosis in tumor tissues were analyzed at 1 h, 5 h, 24 h, 48 h and 336 h after treatment. RESULTS: The inhibitive ratio of treated group was 59.74%, 52.51% and 40.35% respectively. Necrotic areas in tumor tissue were mainly distributing around the magnetic fluid and became larger with the lengthening of treatment. The VEGF expression and apoptosis of the treated group did not show obvious difference from that of the control group at different time points. CONCLUSION: Single time magnetic fluid hyperthermia significantly inhibited growth of grafted H22 tumor in mice, which mainly resulted from inducing necrosis while not inhibiting VEGF expression or inducing apoptosis. PMID- 19219703 TI - Some peculiarities of the sequential action of heat and ionizing radiation on yeast cells. AB - The dependence of the thermal enhancement ratio after a sequential action of heat and ionizing radiation on the dose and dose rate of ionizing radiation as well as on the temperature and duration of its application was studied for yeast cells. The combined effect of heat and ionizing radiation on cell killing depended on both the sequence of application (i.e. whether heat is applied prior to or following irradiation) and the temperature. The effectiveness of treatment with heat and ionizing radiation was greatly dependent on the duration of heat exposure. For an equal amount of cell killing from heat alone, long action of heat (50 degrees C) was more effective for radiosensitization than a short acute action of high heat (58 degrees C). For heating at 50 degrees C, heating after irradiation produced more radiosensitization than heating before irradiation. However, high heating at 58 degrees C before irradiation gave the same radiosensitization as heating after irradiation. These data confirm similar observations for mammalian cells. The results were interpreted by means of a mathematical model in which the synergistic effect of the sequential application of heat and ionizing radiation results from the additional lethal damage arising from the interaction of sublesions induced by both agents. These sublesions are not lethal after the action of these modalities, each taken alone. The model appears to be appropriate and the conclusions are valid. PMID- 19219704 TI - Preoperative chemoradiation combined with regional hyperthermia for patients with resectable esophageal cancer. AB - PURPOSE: To analyse the treatment results of neo-adjuvant chemoradiation combined with regional hyperthermia in patients with resectable esophageal cancer. PATIENTS AND METHODS: Between August 2003 and December 2004, 28 patients entered a phase II study combining chemoradiation over a 4.5-week period with five sessions of regional hyperthermia. Chemotherapy consisted of carboplatin (AUC = 2) and paclitaxel (50 mg/m(2)) and radiotherapy of 41.4 Gy in 1.8 Gy daily fractions. Locoregional hyperthermia was applied using the AMC phased array of four 70 MHz antennas, aiming at a stable tumor temperature of 41 degrees C for one hour. Carboplatin was infused during the hyperthermia session. Esophageal resection was planned at 6-8 weeks after the end of radiotherapy. The majority of the patients had a T3 tumor (86%) and were cN+ (64%). Median follow-up for survivors was 37 months (range 31-46). RESULTS: Twenty-five patients (89%) completed the planned neo-adjuvant treatment and acute toxicity was generally mild. Twenty-six patients were operated on. A pathologically CR, PRmic, PR and SD were seen in 19%, 27%, 31% and 23% respectively. All patients had a R0 resection. In-field locoregional control during follow up for the operated patients was 100%. Quality of life was good for patients without disease progression. Survival rates at one, two and three years were 79%, 57% and 54% respectively. CONCLUSION: Neo-adjuvant chemoradiation combined with regional hyperthermia followed by esophageal resection for patients with esophageal cancer resulted in good locoregional control and overall survival. PMID- 19219705 TI - Radiation exposure to surgical staff during hyperthermic isolated limb perfusion with 99m Technetium labeled red blood cells. AB - PURPOSE: Hyperthermic isolated limb perfusion (HILP) is an effective method in the treatment of recurrent melanomas and soft tissue sarcomas. To avoid systemic toxicity, leakage from the limb perfusate into the systemic circulation is real time monitored by administration of a radioactive agent to the limb circuit. This has made HILP safe for the patient. However, the radiation exposure to the surgical staff has never been measured and could be a limiting factor for the use of HILP. The purpose of the present study was to measure and evaluate the radiation exposure to the surgical staff performing HILP with (99m)Technetium labeled red blood cells. MATERIALS AND METHODS: Thirteen patients had HILP performed in 11 lower limbs and two upper limbs at our inpatient clinic between October 2006 and February 2007. The surgeon and nurse had thermoluminescence dosimetry (TLD) chips attached to the finger pulp and to the ring area of the left fourth finger, as well as an electronic dosimeter attached to the anterior lining of the trousers. The anesthesiologist and perfusion technologist also carried electronic dosimeters. RESULTS: The surgeon had the highest radioactive exposure with an average dose per procedure to the finger pulp of 16.2 microSv, to the ring area of 8.5 microSv, and to the abdominal wall of 4.2 +/- 0.6 microSv. CONCLUSIONS: HILP with (99m)technetium-labeled red blood cells does not constitute a safety risk to the operating team with respect to radioactive exposure. Routine dose monitoring of the staff or special precautions for fertile women are not necessary. PMID- 19219706 TI - More attention, less confusion: time to lessen the burden of delirium. PMID- 19219707 TI - Delirium: a clarion call. AB - Delirium has been historically under studied and under appreciated despite its high frequency and profound impact upon outcomes. This reflects failures at all levels from awareness among frontline staff, ownership by specialist researchers, funding from research bodies, and policy making by senior management and politicians. This is reflected in under detection, suboptimal management and a lack of basic mechanisms by which delirium can be documented within health systems activity data. Not surprisingly, the considerable financial implications are poorly acknowledged and reimbursement arrangements sparse. Delirium as an entity has been victim to its unique position at the mind-body interface and occurrence across multiple disciplines such that no one specialty recognizes delirium as its primary responsibility. It is time for the healthcare community to respond to the challenge of delirium. PMID- 19219708 TI - Eye tracking technology: a fresh approach in delirium assessment? AB - The objective of this paper is to highlight the potential role of eye tracking technology (ETT) in the assessment of delirious patients. Delirium occurs in one in five general hospital admissions (Siddiqi, 2006) and its frequency will increase as society gets older. Despite its frequency and significant independent impact upon morbidity and mortality, delirium remains under studied and is frequently missed, detected late, or misdiagnosed (Farrell & Ganzani, 1995; Inouye, 2001; Kakuma, 2003). Detection is a key target for both clinical and research efforts. Assessment of attention is key to diagnosing delirium, yet nurses and non-research medical staff often fail to correctly identify inattention (Inouye et al., 2001; Lemiengre et al., 2006; Ryan et al., 2008). Eye tracking measures have been used in a plethora of key areas of psychiatric research (Crawford et al., 2005; Corden, Chilvers, & Skuse, 2008; Hardin, Schroth, Pine, & Ernst, 2007; Holzman, Leonard, Proctor, & Hughes, 1973), and provide an accurate and non-invasive method in the assessment of cognitive function. The potential of ETT for direct clinical applications in the assessment of attention and comprehension, key cognitive symptoms of delirium, are promising. This paper considers potential new approaches which recent advancements in non-invasive ETT may bring to the examination and understanding of delirium. PMID- 19219709 TI - The virtual gateway: opening doors in delirium teaching and learning. AB - Delirium is a common, serious and potentially preventable problem. It is argued here that knowledge and skills for effective assessment and treatment of delirium cannot be considered 'specialist'. Rather, delirium is a problem that requires a healthcare team approach to assessing patients, identifying risks or symptoms and reducing precipitating factors. Research from the preceding decade suggests that the clinical reality is falling short of this ideal with the result that many cases of delirium go undetected and many precipitating factors are not reduced where this is indeed possible. This presents a challenge for the education of nursing and medical students and for the development of qualified staff in practice. The current paper outlines an educational approach, e-learning, which increases access for learners, integrates knowledge and skill development and promotes active, problem-based learning. PMID- 19219710 TI - The genetics of deliria. AB - Delirium not induced by alcohol or other psychoactive substance and alcohol withdrawal delirium (or delirium tremens) are both cerebral syndromes with similar presentations and are associated with various adverse outcomes. Recently, interest in identifying genetic predisposing factors that influence the occurrence or the outcome of delirium has become a prominent point of delirium research. We systematically searched published articles concerning genetic associations and the occurrence and outcome of delirium. Of 33 identified articles, six investigated non-alcohol withdrawal delirium, and from those six, five evaluated an association with apolipoprotein E (APOE). One association of APOE genotype with the emergence of delirium and two associations of APOE genotype with the duration of delirium were reported. The remaining 27 identified articles investigated genetic associations with alcohol withdrawal delirium and were mainly related to dopamine. Two studies reported a significant association of alcohol withdrawal delirium with the dopamine transporter gene (SLC6A3) and the dopamine receptor 3 (DRD3). Results are inconclusive, and no hard evidence exists due primarily to insufficiently powered studies and other methodological issues. Prospective studies incorporating systematic and rigorous diagnostic criteria and involving long term follow up are needed to advance understanding of this field. PMID- 19219711 TI - Delirium and long-term cognitive impairment. AB - Delirium is a severe, acute neuropsychiatric syndrome that is highly prevalent in acute hospital populations. Delirium has noticeable effects on length of hospitalization, cost of care, mortality and morbidity. In addition to these well established adverse consequences, there is increasing evidence linking delirium and a higher risk of long-term cognitive impairment (LTCI), including dementia. A prior review (Jackson, Gordon, Hart, Hopkins, & Ely, 2004), in which nine studies (total N = 1,885, years 1989-2003) were considered, concluded that there was evidence for an association between delirium and LTCI. Here we provide a review of studies published since Jackson's review. We included nine reports, with a total of 2,025 patients. The studies show diverse sample sizes, methodologies, designs and patient populations. However, taken together, the results of these new studies broadly confirm that there is a link between delirium and LTCI. We go on to discuss putative mechanisms and explanations. These include (1) delirium as a marker of chronic progressive pathology, but unrelated to any progression, (2) delirium as a consequence of acute brain damage which is also responsible for a 'single hit' or triggering of active processes causing LTCI, (3) delirium itself as a cause of LTCI, and (4) drug treatment of delirium or other conditions as a cause of LTCI. We conclude with suggestions for future research. PMID- 19219712 TI - Delirium in the intensive care unit. AB - Delirium is a common manifestation of acute brain dysfunction in critically ill patients with prevalence as high as 75%. In the last years there has been a progressive increase of publications regarding intensive care (ICU) delirium, acknowledging its importance. The occurrence of delirium in ICU is related to more adverse outcomes including self-extubation and removal of catheters, prolonged hospitalization, increased costs, higher mortality, and potentially, long-term cognitive impairment. The pathophysiology explaining the processes subtending the development of delirium is still elusive, though several theories have been discussed. It is known that different risk factors are associated with delirium in the ICU. Patients in ICU frequently receive medications to treat pain and to ensure sedation, but an association between these drugs and delirium has been shown. Therefore, this pharmacological exposure should be modified to reduce the risk factors. Giving the multifactorial genesis of delirium, multicomponent interventions to prevent delirium developed in non-ICU settings can be adapted to critically ill patients with the purpose of reducing the incidence. When delirium is diagnosed the use of typical and atypical antipsychotics may be effective for its treatment. Future studies should evaluate target interventions to prevent delirium in the ICU. PMID- 19219713 TI - Motor subtypes of delirium: past, present and future. AB - Clinical subtyping of delirium according to motor-activity profile has considerable potential to account for the heterogeneity of this complex and multifactorial syndrome. Previous work has identified a range of clinically important differences between motor subtypes in relation to detection, causation, treatment experience and prognosis, but studies have been hampered by inconsistent methodology, especially in relation to definition of subtypes. This article considers research to date, including a number of recent studies that have attempted to address these issues and identify a means of achieving greater consistency in approaches to subtyping. Possibilities for future work are discussed and a research plan for the field is outlined. PMID- 19219714 TI - Statistical methods for analysing longitudinal data in delirium studies. AB - Delirium is an often acute and highly fluctuating syndrome that can be transient or in some cases associated with prolonged disturbances. The best way to capture its natural course is to conduct studies with longitudinal design, but data analysis in longitudinal studies is difficult, as often the measured variables of each subject are correlated over the course of time. As such, there has been limited application of such methods for analysing longitudinal data in the study of delirium. This overview considers simple traditional approaches along with more complex methods that involve modelling of data. The relative merits of survival analysis, structural equation modelling, and path analysis are reviewed. Furthermore, two flexible modelling techniques are considered; the mixed effects model and generalized estimating equations with emphasis on their use with binary outcomes, as often the outcome in delirium studies is delirium/no delirium. Their contrasting approach to parameter interpretation and methods for accounting for correlation and dealing with missing data are detailed. Information on available software is provided. Delirium research will be substantially enhanced by incorporating such methods. PMID- 19219720 TI - Further studies on CAB approach toward chemical conversion of C19-diterpenoid alkaloids to taxoids: synthesis of the vital intermediate C-nor-aconanone. AB - In the title study, the synthesis of the vital intermediate C-nor-aconanone (3) from 4 was completed through 11 steps, mainly including semipinacol rearrangement, formation of the imines by the treatment of 10 or 20 with NBS, the cleavage of N-C19 bonds in 11 or 21 by treatment with m-CPBA and subsequently with LTA, as well as the rupture of the N-C17 bonds in 22 by a modified Nef reaction (NaH/t-BuOH --> KMnO4/H2O). One-pot procedure was successfully developed starting from 11 or 21 to afford the N,19-seco-C-nor product 15 or 22, respectively, in reasonably good yields. PMID- 19219721 TI - New antioxidant phenylethanol glycosides from Torenia concolor. AB - Two new phenylethanol glycosides, phenylethyl-O-alpha-l-rhamnopyranosyl-(1 --> 2) beta-D-glucopyranoside (torenoside A, 1) and 2'-O-3,4-dihydroxy-beta-phenylethoxy O-alpha-L-rhamnopyranosyl-(1'' --> 3')-(4'-O-caffeoyl)-beta-D-glucopyranoside (torenoside B, 2), along with the 17 known compounds (3-19) were isolated from Torenia concolor. Those structures were established on the basis of spectroscopic analysis including NMR spectroscopic techniques (13C, 1H, 1H-1H COSY, HMQC, HMBC, TOCSY, and NOESY). Moreover, phenylethanol glycosides 3-6 exhibited significant antioxidant activities in DPPH radical scavenging assay. PMID- 19219722 TI - Two new stilbenoids from Pleione bulbocodioides. AB - Two new stilbenoids, 9-(4'-hydroxy-3'-methoxyphenyl)-10-(hydroxymethyl)-11 methoxy-5,6,9,10-tetrahydrophenanthro[2,3-b]furan-3-ol and 2-(4''-hydroxybenzyl) 3-(3'-hydroxyphenethyl)-5-methoxy-cyclohexa-2,5-diene-1,4-dione, together with the three known stilbenoids were isolated from the tubers of Pleione bulbocodioides (Franch.) Rolfe. Their structures were elucidated by spectroscopic methods. PMID- 19219723 TI - Triterpenoid saponins from Anemone flaccida induce apoptosis activity in HeLa cells. AB - Five triterpenoid saponins were isolated from Anemone flaccida Fr. Schmidt. Their structures were identified as glycoside St-I4a (1), glycoside St-J (2), anhuienoside E (3), hederasaponin B (4), and flaccidoside II (5). Compounds 1-2 were isolated from Anemone family for the first time, and compounds 3-4 were isolated from this plant for the first time. The inhibitory effects of saponins on proliferation of HeLa cells were studied by MTT assay, the apoptosis-induction activity was observed by cell-cycle analysis and caspase-3 expression assay. The antitumor activities of the saponins were ranked in the following order: 5 > 3 > 4 > 1 > 2. The data presented here indicated that naturally occurring triterpenoid saponins can be regarded as excellent structures for the potential development of new anticancer agents. PMID- 19219724 TI - Two triterpenes from Lysimachia foenum-graecum. AB - One new oleane-type triterpene saponin, named lysimachiagenoside A and the known 21-O-angeloylbarringtogenol C were isolated from the aerial parts of Lysimachia foenum-graecum Hance. 21-O-angeloylbarringtogenol C was a new natural product. These structures were identified on the basis of 1D- and 2D-NMR techniques, including 1H-1H COSY, HMQC, HMBC, TOCSY, and ROESY experiments as well as chemical methods. PMID- 19219725 TI - Aromatase inhibitory fatty acid derivatives from the pollen of Brassica campestris L. var. oleifera DC. AB - Two new fatty acid derivatives, 9Z,12Z,15Z-octadecatrienoic acid sorbitol ester (1) and (10,11,12)-trihydroxy-(7Z,14Z)-heptadecadienoic acid (2), were isolated from the pollen of Brassica campestris L. var. oleifera DC., along with the four known fatty acid derivatives, 9Z,12Z,15Z-octadecatrienamide, N-(2-hydroxyethyl) (3), hexadecanoic acid sorbitol ester (4), 15,16-dihydroxy-9Z,12Z-octadecadienoic acid (5), and 9Z,12Z,15Z-octadecatrienoic acid 2,3-dihydroxypropyl ester (6). Their structures were elucidated by extensive spectroscopic analysis, including 1D- and 2D-NMR as well as HR-ESI-MS experiments. All compounds were tested using a noncellular aromatase assay, and the results showed that some compounds possessed strong inhibitory activity. PMID- 19219726 TI - Two new dimeric stilbenes from the stem bark of Morus australis. AB - Two new dimeric stilbenes austrafuran B and austrafuran C were isolated from the bark of Morus australis. Their structures were elucidated on the basis of spectroscopic methods. PMID- 19219727 TI - Pseudolaric acid B-induced apoptosis through p53-dependent pathway in human gastric carcinoma cells. AB - Pseudolaric acid B (PLAB, 1), a natural diterpenoid compound, was isolated from Pseudolarix kaempferi Gordon. It has shown antifungal, antifertility, and antiangiogenic properties in previous studies. Recently, increasing evidence has confirmed that 1 exhibits antitumor effects in several tumor cell lines, but the underlying mechanism has not been fully elucidated. The aim of this study was to investigate the mechanism of PLAB-induced cell apoptosis in MGC803 cells. The results showed that 1 significantly inhibited the proliferation of MGC803 cells at 0.01-10 microM and the IC(50) value was 0.91 microM for 48 h. PLAB-induced apoptosis in MGC803 cells was confirmed by DNA fragmentation assay and Hoechst33342/PI staining. PLAB-treated MGC803 cells were arrested at G(2) phase, which was associated with a marked increment of the expression of cyclin dependent kinase inhibitor p21. The induction of p21 appeared to be transcriptionally up-regulated and was p53-dependent. In addition, PLAB induced Fas/APO-1 and caspase-3 expressions that were also correlated with apoptosis. Meanwhile, 1 decreased the mRNA expression of bcl-2, which is an antiapoptosis factor. In conclusion, 1 induced apoptosis through p53-dependent pathway in human gastric carcinoma cells. These findings suggest that 1 may be a novel promising agent for treating human gastric carcinoma. PMID- 19219728 TI - Two new Daphniphyllum alkaloids from Daphniphyllum macropodum Miq. AB - Two new Daphniphyllum alkaloids, 4,21-deacetyl-deoxyyuzurimine (1) and macropodumine L (2), together with the two known related alkaloids, have been isolated from the bark of Daphniphyllum macropodum Miq. The structures of the new compounds were elucidated on the basis of the detailed analysis of spectroscopic data, chemical method, and by comparison of the spectroscopic data with those of known compounds. Compounds 2 and 4 exhibited weak cytotoxicity against human carcinoma cell lines SMMC-7721 and HO-8910. PMID- 19219729 TI - An investigation of the kinetic and anti-angiogenic properties of plant glycoside inhibitors of thymidine phosphorylase. AB - We investigated the potential of symplocomoside (1) and symponoside (2), glycosides isolated from the bark of Symplocos racemosa to inhibit thymidine phosphorylase (TP) activity and associated angiogenesis. Compound 1 was a reversible, noncompetitive inhibitor of deoxythymidine binding to TP (IC(50) = 65.45 +/- 5.08 microM; K(i) = 62.83 +/- 2.10 microM) and 2 was a reversible, uncompetitive inhibitor (IC(50) = 94.17 +/- 4.05 microM; K(i) = 101.95 +/- 1.65 microM). Molecular modeling analysis indicated that both compounds bound at the active site of the enzyme but not solely to amino acid residues involved in catalysis. Both compounds were active in in vitro angiogenic assays inhibiting endothelial cell migration and invasion in Matrigel, but did not inhibit growth factor-induced proliferation and were not cytotoxic. Compound 1 may have potential as an anti-angiogenic and anti-tumor agent. PMID- 19219730 TI - Chemical constituents of the stems of Ephedra sinica. AB - A new naphthalene derivative, 1-methyl-2,3-methylenedioxy-6-naphthalenecarboxylic acid methyl ester (1), and a new alkaloid, ( +/- )-1-phenyl-2-imido-1-propanol (2), together with the four known compounds, ephedrine, pseudoephedrine, N methylephedrine, and 6-methoxykynurenic acid, have been isolated from the stems of Ephedra sinica, a famous traditional Chinese herbal medicine. The structures of 1 and 2 were determined by spectroscopic methods, including 1D- and 2D-NMR experiments as well as HR-EI-MS analysis. PMID- 19219731 TI - The synthesis of analogs of shuangkangsu, a novel natural cycloperoxide glucoside from Lonicera japonica Thunb. AB - Four novel optically pure cycloperoxide glucosides 9a, 9b, 10a, and 10b, analogs of shuangkangsu--a natural product with unusual skeleton and antivirus activity from the buds of Lonicera japonica Thunb, were firstly synthesized by employing peroxidation and glucosidation reactions from phthalaldehyde or 4,5-dichloro phthalaldehyde and glucose. PMID- 19219732 TI - C21 steroidal glycosides from Cynanchum wallichii Wight. AB - Three new C21 steroidal glycosides, characterized with rostratamine and qinyangshengenin as aglycone moiety and 2,6-dideoxy-pyranoses as component sugars, have been isolated from the roots of Cynanchum wallichii Wight. The structures of the three new C21 steroidal glycosides were elucidated as rostratamine-3-O-beta-D-oleandropyranosyl-(1 --> 4)-beta-d-cymaropyranosyl-(1 --> 4)-beta-D-cymaropyranoside, qinyangshengenin-3-O-beta-D-cymaropyranosyl-(1 --> 4) beta-D-digitoxo-pyranoside, and qinyangshengenin-3-O-beta-D-oleandropyranosyl-(1 -> 4)-beta-D-cymaropyranosyl-(1 --> 4)-beta-D-digitoxopyranoside. PMID- 19219733 TI - A new acylated flavone glycoside from Colebrookea oppositifolia. AB - A new acylated flavone glycoside, echioidinin 2'-O-beta-D-(2''-O acetyl)glucopyranoside (1) along with the three known flavonoids, 5,6,7,8,5' pentamethoxy-3',4'-methylenedioxyflavone (2), 5,2',6'-trihydroxy-7-methoxyflavone (3), and kaempferol 7,4'-dimethyl ether 3-O-beta-D-glucopyranoside (4), were isolated from the roots of Colebrookea oppositifolia. The structure of compound 1 was elucidated by extensive spectral and chemical studies. PMID- 19219734 TI - Microbial transformation of dehydroandrographolide by Cunninghamella elegans. AB - The biotransformation of dehydroandrographolide (1) by Cunninghamella elegans was performed and four transformed products were obtained. Based on their extensive spectral data, the structures of these metabolites were identified as 3-oxo dehydroandrographolide (2), 3-oxo-2beta-hydroxy-dehydroandrographolide (3), 3-oxo 8beta,17alpha-epoxydehydroandrographolide (4), 3,19-dihydroxy-7,11,13-ent labdatrien-15,16-olide (5), respectively. Among them, products 3-5 are new compounds. PMID- 19219735 TI - A new cytotoxic homoisoflavonoid from Dracaena cambodiana. AB - A new homoisoflavonoid, named cambodianol (1), together with the two known flavanes, (2S)-7,3'-dihydroxy-4'-methoxy-8-methylflavane (2) and (2R)-7,4' dihydroxy-8-methylflavane (3), were isolated from the stems of Dracaena cambodiana. Their structures were determined based on HR-ESI-MS and spectroscopic techniques (UV, IR, 1D-, and 2D-NMR). Compound 1 exhibited significant cytotoxic activities against K562 and SGC-7901 with the IC(50) values of 1.4 and 2.9 microg/ml, respectively. PMID- 19219736 TI - Variability and progression in competitive performance of Paralympic swimmers. AB - Analysis of variability and progression in performance of top athletes between competitions provides information about performance targets that is useful for athletes, practitioners, and researchers. In this study, 724 official finals times were analysed for 120 male and 122 female Paralympic swimmers in the 100-m freestyle event at 15 national and international competitions between 2004 and 2006. Separate analyses were performed for males and females in each of four Paralympic subgroups: S2-S4, S5-S7, S8-S10 (most through least physically impaired), and S11-S13 (most through least visually impaired). Mixed modelling of log-transformed times, with adjustment for mean competition times, was used to estimate variability and progression. Within-swimmer race-to-race variability, expressed as a coefficient of variation, ranged from 1.2% (male S5-S7) to 3.7% (male S2-S4). Swimming performance progressed by approximately 0.5% per year for males and females. Typical variation in mean performance time between competitions was approximately 1% after adjustment for the ability of the athletes in each competition, and the Paralympic Games was the fastest competition. Thus, taking into account variability, progression, and level of competition, Paralympic swimmers who want to increase substantially their medal prospects should aim for an annual improvement of at least 1-2%. PMID- 19219737 TI - Synthesis of oligonucleotides containing 2'-deoxyguanosine adducts of nitropyrenes. AB - Two different approaches to synthesize oligonucleotides containing the 2 ' deoxyguanosine adducts formed by nitropyrenes are described. A direct reaction of an unmodified oligonucleotide with an activated nitropyrene derivative is a convenient biomimetic approach for generating the major adducts in DNA. A total synthetic approach, by contrast, involves several synthetic steps, including Buchwald-Hartwig Pd-catalyzed coupling, but can be used for incorporating both the major and minor adducts in DNA in high yield. PMID- 19219739 TI - Synthesis of zidovudine derivatives with anti-HIV-1 and antibacterial activities. AB - Twelve novel zidovudine derivatives were prepared by modifying 5 '-hydroxyl group of sugar moiety (1-8) and 5-methyl group of thymidine nucleus (9-12) and characterized spectrally. The compounds were evaluated for anti-HIV-1, antitubercular and antibacterial activities. Compound (3-azido-tetrahydro-5- (3,4 dihydro-5-methyl-2,4-dioxopyrimidin-1(2H)-yl)furan-2-yl)methyl 7-(4-(2 phenylacetoyloxy)-3,5- dimethylpiperazin-1-yl)-5-(2-phenylacetoyloxyamino)-1 cyclopropyl-6,8-difluoro-1,4-dihydro-4-oxoquinoline-3-carboxylate (5) was found to be the most potent anti-HIV-1 agent with EC(50) of 0.0012 microM against HIV 1(IIIB) and CC(50) of 34.05 microM against MT-4 with selectivity index of 28,375. Compound 5 inhibited Mycobacterium tuberculosis with MIC of 1.72 microM and inhibited four pathogenic bacteria with MIC of less than 1 microM. PMID- 19219738 TI - Synthesis and evaluation of an RNA editing substrate bearing 2'-deoxy-2' mercaptoadenosine. AB - The RNA-editing adenosine deaminases (ADARs) catalyze deamination of adenosine to inosine in double stranded structure found in various RNA substrates, including mRNAs. Here we describe the synthesis of a phosphoramidite of 2'-deoxy-2' mercaptoadenosine and its incorporation into an ADAR substrate. Surprisingly, no deamination product was observed with this substrate indicating replacing the 2' OH with a 2'-SH at the editing site is highly inhibitory. Modeling of nucleotide binding into the active site suggests the side chain of T375 of human ADAR2 to be in proximity of the 2'-substituent. Mutation of this residue to cysteine caused a greater that 100-fold reduction in deamination rate with the 2'-OH substrate. PMID- 19219740 TI - Novel acyclic amide-conjugated nucleosides and their analogues. AB - An effective one-step synthesis of new amide-conjugated nucleosides and their analogues, in the presence of 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4 methylmorpholinium chloride (DMT-MM) as the condensing agent, is presented. Substrate subunits carrying carboxylic group were obtained by acidic hydrolysis of Michael-type adducts of various 5-substituted uracil derivatives to methyl acrylate. Amine substrate was synthesized by reduction of 1-(2' cyanoethyl)thymine with sodium borohydride in the presence of nickel (II) chloride as catalyst. Other applied amine substrates were 5'-amino-5' deoxythymidine and 5-aminouracil. PMID- 19219742 TI - Short synthesis and antiviral activity of acyclic phosphonic acid nucleoside analogues. AB - An efficient route for synthesizing novel allylic and cyclopropanoid phosphonic acid nucleoside analogues is described. The condensation of the bromine derivatives 6 and 18 with nucleoside bases (A, U, T, C, G) under standard nucleophilic substitution and deprotection conditions, afforded the target phosphonic acid nucleoside analogues. These compounds were evaluated for their antiviral properties against various viruses. Cyclopropanoid phosphonic adenine nucleoside analogue 23 showed significant anti-HIV activity. PMID- 19219741 TI - Reactions of peroxynitrite with uric acid: formation of reactive intermediates, alkylated products and triuret, and in vivo production of triuret under conditions of oxidative stress. AB - Hyperuricemia is associated with hypertension, metabolic syndrome, preeclampsia, cardio-vascular disease and renal disease, all conditions associated with oxidative stress. We hypothesized that uric acid, a known antioxidant, might become prooxidative following its reaction with oxidants; and, thereby contribute to the pathogenesis of these diseases. Uric acid and 1,3-(15)N(2)-uric acid were reacted with peroxynitrite in different buffers and in the presence of alcohols, antioxidants and in human plasma. The reaction products were identified using liquid chromatography-mass spectrometry (LC-MS) analyses. The reactions generate reactive intermediates that yielded triuret as their final product. We also found that the antioxidant, ascorbate, could partially prevent this reaction. Whereas triuret was preferentially generated by the reactions in aqueous buffers, when uric acid or 1,3-(15)N(2)-uric acid was reacted with peroxynitrite in the presence of alcohols, it yielded alkylated alcohols as the final product. By extension, this reaction can alkylate other biomolecules containing OH groups and others containing labile hydrogens. Triuret was also found to be elevated in the urine of subjects with preeclampsia, a pregnancy-specific hypertensive syndrome that is associated with oxidative stress, whereas very little triuret is produced in normal healthy volunteers. We conclude that under conditions of oxidative stress, uric acid can form reactive intermediates, including potential alkylating species, by reacting with peroxynitrite. These reactive intermediates could possibly explain how uric acid contributes to the pathogenesis of diseases such as the metabolic syndrome and hypertension. PMID- 19219743 TI - Characterization of human cytochrome P450 enzymes involved in the biotransformation of eperisone. AB - Eperisone is a centrally acting muscle relaxant widely used for the therapeutic treatment of spastic patients to relieve muscle stiffness and back pain. The objective of this study was to characterize the metabolic pathway involved in the biotransformation of eperisone mediated by human cytochrome P450 (CYP) enzymes. Eperisone was metabolized to seven metabolites via oxidation and carbonyl reduction in human liver microsome. Among them, M3 and M4 were found to be primary major metabolites which were generated by CYPs. The kinetics study with ( )-R- and (+)-S-eperisones revealed that CYPs-mediated hydroxylation did not have significant stereoselectivity for metabolic clearance of eperisone. Incubation with recombinant CYP isozyme, chemical inhibition assay, and immuno-inhibition assay showed that multiple CYPs were involved in M4 formation, but mainly CYP2J2 in M3 formation. In addition, intestinal microsomes metabolized eperisone to M3 and M4 via CYP2J2- and CYP3A4-mediated reactions, which are supposed to contribute to presystemic metabolism of eperisone. PMID- 19219744 TI - Acetaminophen bioactivation by human cytochrome P450 enzymes and animal microsomes. AB - Acetaminophen is a widely used analgesic antipyretic agent. When used at low doses, it is a safe drug, but at higher doses it can cause acute hepatic necrosis in humans and experimental animals. The key mechanism in the hepatotoxicity is cytochrome P450 (CYP)-catalysed formation of the reactive metabolite, N-acetyl-p benzoquinone imine (NAPQI) that is capable of binding to cellular macromolecules and in that way an LC/MS liquid chromatography/mass spectrometry (LC/MS) method was developed to measure NAPQI formation by trapping it to reduced glutathione. This method was used to determine the bioactivation of acetaminophen at two concentrations: 50 microM therapeutic and 1 mM toxic by using nine human recombinant CYP enzymes: CYP1A1, CYP1A2, CYP2A6, CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A4; and with different microsomes from experimental animals. At the toxic concentration the formation of NAPQI-glutathione was highest with CYP3A4 followed by CYP2E1, CYP1A2, and CYP2D6. At the therapeutic concentration, CYP3A4 had also the highest bioactivation capacity. In a comparison of the enzyme kinetics, CYP3A4 was the most efficient CYP with the lowest K(m) value 130 microM (95% confidence interval = 63-210 microM). Dexamethasone-induced rat liver microsomes had the most effective bioactivation capacity at therapeutic and toxic acetaminophen concentrations. This study suggests that CYP3A4 is the major CYP enzyme form catalysing acetaminophen oxidation to NAPQI in human liver. PMID- 19219745 TI - Development of an optimized procedure for the preparation of rat intestinal microsomes: comparison of hepatic and intestinal microsomal cytochrome P450 enzyme activities in two rat strains. AB - The objective of this study was to characterize cytochrome P450 (CYP) activities in both intestinal and hepatic microsomes from Wistar and Sprague-Dawley rats. Specific probes for measuring CYP activities were selected using rat recombinant CYP. The intestinal microsome preparation was optimized getting a more relevant and reproducible abundance of CYPs to measure CYP activities. Testosterone, propranolol, diclofenac, and midazolam were determined as specific substrates of rat CYP2C11, CYP2D2, CYP2C6, and CYP3A, respectively. Ethoxyresorufin and pentoxyresorufin were not specific substrates of CYP1A2 and CYP2B1, respectively. Hepatic and intestinal microsomes expressed active CYP1A1, CYP1A2, CYP2B1, and CYP3A2. Only liver expressed active CYP2C6, CYP2C11, and CYP2D2. Wistar liver expressed more active CYP1A and CYP3A2, but less active CYP2B1 than Wistar intestine. Sprague-Dawley liver expressed more active CYP2B1 and CYP3A2, but less active CYP1A than Sprague-Dawley intestine. In conclusion, CYP activities were qualitatively equivalent but not quantitatively in both strains. PMID- 19219746 TI - Preclinical pharmacokinetics of MK-0974, an orally active calcitonin-gene related peptide (CGRP)-receptor antagonist, mechanism of dose dependency and species differences. AB - The underlying mechanism for low oral bioavailability of MK-0974, a potent calcitonin-gene related peptide (CGRP)-receptor antagonist, in monkeys and for species-dependent non-linear pharmacokinetics in monkeys and rats were investigated. In monkeys, MK-0974 displayed moderate clearance (14-20 ml min(-1) kg(-1)), while oral bioavailability was 6%. The pharmacokinetics of MK-0974 remained linear across 0.5-10 mg kg(-1) intravenous dose in monkeys, but the oral area under the plasma concentration-time curve (AUC) increase (5-30 mg kg(-1)) was 15-fold over dose-proportional. Based on a comparison of AUC following hepatic portal vein administration and cephalic vein infusion, MK-0974 exhibited a low-to-moderate hepatic extraction ratio (36%) in monkeys. Following oral dose of [14C]MK-0974 to monkeys, the hepatic portal AUC ratio of MK-0974 versus total radioactivity was 0.32, and the total radioactivity recovered in bile and urine was 45-83%. MK-0974 undergoes significant oxidative metabolism (cytochrome P450 (CYP) 3A) in monkey intestinal microsomes. In contrast, oral AUC of MK-0974 in rats was near dose-proportional (15-100 mg kg(-1)). Following oral administration of [14C]MK-0974 to rats, the hepatic portal AUC ratio of MK-0974 to total radioactivity (0.67) was higher than in monkeys. Additionally, the metabolic rate of MK-0974 was slower in rat than in monkey intestinal microsomes. Collectively, intestinal first-pass metabolism played a significant role in the low oral bioavailability in monkeys and contributed to the species-dependent non-linear oral pharmacokinetics in rats and monkeys of MK-0974. PMID- 19219747 TI - Absorption, distribution, metabolism and excretion (ADME) study with 2,2',4,4',5,6'-hexabromodiphenyl ether (BDE-154) in male Sprague-Dawley rats. AB - A metabolism study of orally administered 2,2',4,4',5,6'-hexabromodiphenyl ether (BDE-154; 11.3 micromoles kg(-1)) was conducted in conventional and bile duct cannulated male Sprague-Dawley rats. In conventional rats, approximately 31% of the radiolabelled dose was retained at 72 h, and lipophilic tissues were the preferred sites for disposition. Urinary excretion of BDE-154 was very low (1.0%), and parent compound was detected. Cumulative biliary excretion was 1.3%, and glutathione conjugates were suggested. Over 62% of the dose in conventional male rats was excreted in faeces, and was composed of parent compound (7.3%), free metabolites (13.1%), and covalently bound residues (41.4%). Faecal metabolites characterized by gas chromatography/mass spectrometry included multiple isomers of monohydroxylated hexa-/penta-/tetrabromodiphenyl ethers, and di-hydroxylated hexa/pentabromodiphenyl ethers. The adipose tissue 14C was extractable BDE-154, but 40% of liver 14C was bound to macromolecules. The study demonstrated the importance of performing individual polybrominated diphenyl ether (PBDE) metabolism studies to understand fully PBDE pharmacokinetics. PMID- 19219748 TI - Presentation of a modified dispersion model (MDM) for hepatic drug extraction and a new methodology for the prediction of the rate-limiting step in hepatic metabolic clearance. AB - The main objectives were to investigate the roles of and interplay between determinants of hepatic clearance (CL(H)) in humans, to develop a methodology and reference system for the evaluation and prediction of the rate-limiting step in CL(H), and to update the dispersion model and compare it with traditionally used liver extraction models. The new methodology enables predictions of the hepatic uptake and CL(H), dissociation, and rate-limiting step. In general, absorption, dissociation and diffusion are comparably rapid processes, and metabolism is rate limiting. The liver appears to have a high passive uptake capacity. The Modified Dispersion Model (MDM) has a dispersion number of 0.5 and a distribution factor (df = 0.87) for the correction of a longer hepatic transit time of unbound molecules and the exclusion of the hidden unbound fraction within erythrocytes. Liver models are functionally equivalent at low CL(H), but differ for highly extracted compounds. Well-stirred and parallel-tube models demonstrate the greatest difference in performance, for example, 6- and 800,000-fold differences in the estimated in vivo intrinsic CL(H) and predicted oral bioavailability of the high CL(H) drug naloxone, respectively. The roles of and interplay between determinants of CL(H) have been further clarified and can now be better predicted. Apparent advantages with the MDM include its scientific rationale and intermediate/ balanced performance. PMID- 19219749 TI - Trans-resveratrol-mediated inhibition of beta-oestradiol conjugation in MCF-7 cells stably expressing human sulfotransferases SULT1A1 or SULT1E1, and human liver microsomes. AB - High concentrations of endogenous oestradiol (E2) correlate with the proliferation of cancer cells. Resveratrol (a dietary chemopreventive agent) at high concentrations has an anti-oestrogenic effect. E2 and resveratrol are conjugated via common uridine diphosphoglucuronosyltransferase (UGT) and sulfotransferases (SULT) enzymes. Experiments were conducted in MCF-7 mammalian cells stably expressing human SULT1A1 or SULT1E1 to observe the effect of resveratrol on E2-mediated cell proliferation. The combination of E2 and resveratrol did have a proliferative effect in cells expressing SULT1E1, but not in those expressing SULT1A1. The effect of resveratrol (1-500 microM) on the glucuronidation of E2 (0.25-2.25 microM) was characterized in human liver microsomes. The highest resveratrol concentration significantly decreased the intrinsic clearance of E2 glucuronidation. The results corroborate the reported significant inhibition of SULT1E1-mediated E2 sulfation in vitro by resveratrol. Thus, resveratrol may interact with E2 in vivo by inhibiting its conjugation. PMID- 19219750 TI - Mechanisms of action of two different natural mixtures of persistent organic pollutants (POPs) in ovarian follicles. AB - The present work investigated the effects of two different natural mixtures on aryl hydrocarbon receptor (AhR) and oestrogen receptor (ER)beta protein levels, as well as on the activity of cytochrome P450 (CYP) 1A1 and CYP2B. Consequently, the authors observed the effects of these mixtures on gonadotropine-stimulated steroid secretion by ovarian follicles. The natural mixtures that were studied were 'Mjosa' extracted from burbot liver, which contains a high level of PBDEs, and 'Marine mix', extracted from Atlantic cod liver, which contains a high level of polychlorinated biphenyls (PCBs). Follicular cells were exposed in vitro to 'Marine mix' and 'Mjosa mix' at doses of 3.6 and 1.4 microg ml(-1), respectively. Media were collected and used for steroid analysis and cell viability assays. Cells were used to estimate aromatase activity (CYP19), AhR and ER protein levels, and CYP1A1 and CYP2B1 activity. Western blot analysis indicated down regulation of AhR by 'Marine mix' and down-regulation of ERbeta by Mjosa mix. Up regulation of CYP1A1 expression and activity were seen following treatment with Marine mix, but not Mjosa mix. Increased CYP2B1 activity was noted after treatment with both 'Marine mix' and Mjosa mix. Both mixtures increased luteinizing hormone (LH)-stimulated progesterone and testosterone secretion, follicle-stimulating hormone (FSH)-stimulated oestradiol secretion, and CYP19 activity. These results suggest that: (1) 'Marine mix' is a mixed-type CYP inducer; (2) 'Mjosa mix' is an inducer of ERbeta and CYP2B; and (3) both 'Marine mix' and 'Mjosa mix' stimulate aromatase activity as a consequence of oestradiol secretion through activation of CYP19. PMID- 19219752 TI - Applying an exemplar model to the serial reaction-time task: anticipating from experience. AB - We present a serial reaction time (SRT) task in which participants identified the location of a target by pressing a key mapped to the location. The location of successive targets was determined by the rules of a grammar, and we varied the redundancy of the grammar. Increasing both practice and the redundancy of the grammar reduced response time, but the participants were unable to describe the grammar. Such results are usually discussed as examples of implicit learning. Instead, we treat performance in terms of retrieval from a multitrace memory. In our account, after each trial, participants store a trace comprising the current stimulus, the response associated with it, and the context provided by the immediately preceding response. When a target is presented, it is used as a prompt to retrieve the response mapped to it. As participants practise the task, the redundancy of the series helps point to the correct response and, thereby, speeds retrieval of the response. The model captured performance in the experiment and in classic SRT studies from the literature. Its success shows that the SRT task can be understood in terms of retrieval from memory without implying implicit learning. PMID- 19219751 TI - Statin-induced myopathy in the rat: relationship between systemic exposure, muscle exposure and myopathy. AB - Rare instances of myopathy are associated with all statins, but cerivastatin was withdrawn from clinical use due to a greater incidence of myopathy. The mechanism of statin-induced myopathy with respect to tissue disposition was investigated by measuring the systemic, hepatic, and skeletal muscle exposure of cerivastatin, rosuvastatin, and simvastatin in rats before and after muscle damage. The development of myopathy was not associated with the accumulation of statins in skeletal muscle. For each statin exposure was equivalent in muscles irrespective of their fibre-type sensitivity to myopathy. The low amount of each statin in skeletal muscle relative to the liver does not support a significant role for transporters in the disposition of statins in skeletal muscle. Finally, the concentration of cerivastatin necessary to cause necrosis in skeletal muscle was considerably lower than rosuvastatin or simvastatin, supporting the concept cerivastatin is intrinsically more myotoxic than other statins. PMID- 19219753 TI - Frontal and parietal lobe involvement in the processing of pretence and intention. AB - We assessed whether different processes might be at play during pretence understanding by examining breakdowns of performance in participants with acquired brain damage. In Experiment 1 patients with frontal or parietal lesions and neurologically intact adults were asked to categorize videos of pretend and real actions. In Experiment 2 participants saw three types of videos: real intentional actions, real accidental actions, and pretend actions. In one session they judged whether the actions they saw were intentional or accidental, and in a second session they judged whether the actions were real or pretend. Parietal patients had particular difficulties in the identification of pretend actions, and both parietal and frontal patients were more impaired than controls in understanding the intentional nature of pretence. Analyses of individual patients' performance revealed that parietal lesions, and in particular lesions to the temporo-parietal junction, impaired the ability to discriminate pretend from real actions. However, this did not necessarily affect the discrimination of intentional from unintentional actions, which instead may be independently disrupted by damage to frontal areas. Moreover, spared ability to discriminate pretend actions from real actions, and intentional actions from accidental actions, did not grant a full conceptual understanding of the intentional nature of pretence. The implications for pretence understanding are discussed. PMID- 19219754 TI - [Positron-emission tomography (PET) and PET/CT in head and neck cancer - recommendations of an interdisciplinary consensus conference]. AB - BACKGROUND: Today's available therapeutic options in head and neck cancer patients have led to better treatment modalities tailored to the individually clinical staging of the patients towards a risk adapted tumour management. This, however, is only possible with an accurately pretherapeutic diagnostic regimen and closely posttherapeutic follow-up. METHODS: These issues were discussed by nuclear medicine experts, otorhinolaryngologists, oral surgeons, radiologists, radio-oncologists and oncologists in a meeting that took place in Portschach, Austria, on 05 May 2006. The aim was to discuss the impact and indications of performing FDG PET/CT in patients with head and neck cancer and to outline possible future perspectives. RESULTS: FDG PET/CT is recommended for a better pretherapeutic staging in stage IV according to UICC and should be the method of choice in CUP with lymph node metastases. FDG PET/CT should be performed 3 - 4 months after radiation-/radiochemotherapy to diagnose viable tumour and to avoid false positive results. To evaluate the position and effectiveness of FDG PET/CT in therapy-monitoring further studies are needed. In case of radiation therapy FDG PET/CT allows a tailored treatment of patients with an accurate design of the target volume to reduce damage to the surrounding tissues. CONCLUSIONS: The interdisciplinary consensus reached by the experts is not intended to recommend standard guidelines in the management of head and neck cancer but to summarise and stress the impact of FDG PET/CT on the basis of the present literature and current clinical practise. PMID- 19219756 TI - [The extent of patient consent and reimbursement claim by the head physician in accepting personal treatment responsibility]. PMID- 19219757 TI - [Dental care in case of head and neck cancer - part ii: resection]. AB - The dental care in case of head and neck cancer resection should be accomplished by skilled dentists with focus in prosthetics in cooperation with special skilled dental technicians and a specialist in epithetics. The patient must be introduced to the dentist before operation. Otherwise reconstruction is much more difficult and a gentle treatment for the patient not possible. The dentist can recommend the extraction of teeth intraoperative, which are not necessary for dental rehabilitation. Furthermore a perfect model made from dental imprints before operation is very useful in every step of reconstruction. Concerted sessions including the patient, the surgeon, the dentist and if necessary the epithetic specialist before and after operation would be desireable but often not practicable in daily routine. PMID- 19219759 TI - Pharmacological exploration of the sedative mechanism of hesperidin identified as the active principle of Citrus sinensis flowers. AB - The infusion of flowers of several species of Citrus genera is used as a sedative to treat insomnia in Mexican traditional medicine. The aims of this study were to investigate the sedative effect of different extracts of flowers of Citrus sinensis (L.) Osbeck (Rutaceae) and describe the pharmacological action mechanism of the sedative active compounds of this plant. The methanol and dichloromethane extracts, obtained from the flowers of Citrus sinensis (L.) Osbeck (Rutaceae), showed a dose-dependent sedative effect in the exploratory cylinder model in mice, with ED50 (ip) values of 47.04+/-12.03 mg/kg and 129.15+/-21.25 mg/kg, respectively. Hesperidin (ED50=11.34+/-2.48 mg/kg) was identified in the methanol extract as the sedative active principle of this plant. The pre-treatment with atropine (1 mg/kg I. P.), flumazenil (2 mg/kg I. P.), clonidine (0.01 mg/kg I. P.), isoproterenol (0.3 mg/kg I. P.), haloperidol (0.3 mg/kg I. P.), WAY 100 635 (3 mg/kg I. P.), P-chlorophenylalanine (250 mg/kg I. P., twice per day for 2 days), forskolin (3 mg/kg I. P.) and rolipram (0.173 mg/kg I. P.) did not modify the sedative effect of 30 mg/kg hesperidin. However, the sedative effect of this compound was potentiated by yohimbine (1.25 mg/kg I. P.) and buspirone (1 mg/kg I. P.), and reverted by pretreatment with aminophylline (30 mg/kg I. P.), caffeine (30 mg/kg I. P.) and several doses of 1,3-dimethyl-8-phenylxanthine (10, 30 and 54.7 mg/kg I. P.). These results suggest that adenosine receptors might be involved in the sedative action of hesperidin, identified as the active principle of the flowers of Citrus sinensis. PMID- 19219758 TI - In vivo wound healing activity of the methanolic extract and its isolated constituent, gulonic acid gamma-lactone, obtained from Grewia tiliaefolia. AB - Grewia tiliaefolia is a subtropical tree, its stem bark is widely used in traditional Indian medicines to heal chronic wounds, gastric ulcers, burning sensation, itching and other allergic ailments. Bioassay-directed fractionation and chromatography of the methanolic extract of G. tiliaefolia stem bark has resulted in the isolation of gulonic acid gamma-lactone. The methanolic extract and the isolated constituent were studied for their potency on three different cutaneous wound models, VIZ., excision, incision and dead space wounds in Wistar rats. In the excision wound model, healing was assessed by the rate of wound contraction and period of epithelisation. In the incision wound model, the degree of healing was analysed by determining the skin breaking strength. In the dead space wound model, the parameters used to confirm the healing process were weight of granulation tissue, its tensile strength, hydroxyproline content and histological studies. The extract as well as the constituent demonstrated wound healing activity. Topical application of gulonic acid gamma-lactone (0.2% w/w ointment) caused faster epithelialisation with 94.02% wound contraction on day 16 post-wounding, while in control animals the duration of healing was extended up to 22 days with 79.53% wound contraction. The tensile strength of the incision wound was significantly increased (561.12 +/- 5.18 g) compared to the control (327.63 +/- 6.37 g). In the dead space wound model, a significant increase in weight, tensile strength and hydroxyproline content of the granuloma tissue was observed following oral administration of gulonic acid gamma-lactone (60 mg/kg). Histology of the granuloma tissue showed increased collagenation and the absence of monocytes. The wound healing effect was compared with that of the standard skin ointment nitrofurazone. The results of this investigation provide supportive scientific evidence for the medicinal use of G. tiliaefolia for healing of cutaneous wound. PMID- 19219760 TI - Antioxidant lignans from Mananthes patentiflora. AB - A new tetrahydrofuranoketone lignan (1) and two new tetrahydrofuranoketone lignan glycosides (2, 3) were isolated from the ethanol extract of the aerial part of Mananthes patentiflora. The structure elucidations of these compounds were mainly established on the basis of 1D, 2D NMR and HR-mass spectroscopic analysis. All three compounds revealed potent antioxidant activities within the concentration range tested (0.08-50.0 microM). PMID- 19219761 TI - Lipoma-induced colon intussusception. PMID- 19219762 TI - Giant appendiceal mucocele mimicking gastrointestinal stromal tumor of the cecum. PMID- 19219763 TI - "Short" double balloon enteroscope for endoscopic retrograde cholangiopancreatography with conventional sphincterotomy and metallic stent placement after Billroth II gastrectomy. PMID- 19219764 TI - EUS-guided drainage of ruptured liver abscess. PMID- 19219765 TI - Appendiceal intussusception due to a fecalith mimicking a submucosal tumor. PMID- 19219766 TI - Spraying n-butyl-2-cyanoacrylate (Histoacryl) might be a simple and final technique for bleeding gastrointestinal lesions. PMID- 19219767 TI - [Substitution of inhalation devices in the pharmacy according to a frame contract (dated 1st April, 2008) and individual discount regulations]. PMID- 19219768 TI - [Exercise testing by spiroergometry in a patient with sarcoidosis stage II. An unusual combination of functional impairments]. AB - Sarcoidosis is a multisystem disease of unknown origin characterised by epitheloid cell granulomas in the lung and many other organs. The measurement of the alveolo-arterial O(2)-gradient (AaDO(2)) during exercise is the most sensitive method to detect the involvement of the lung with impairment of gas exchange. The method is suitable for follow-up observations to make the indication for a steroid therapy in the case of deterioration. In this case presentation using the 9-panel display proposed by Wasserman we diagnosed besides a slight impairment of the gas exchange a cardiocirculatory dysfunction with a reduction of the oxygen pulse and a reduced minute ventilation with a decrease of the alveolar ventilation. The hypoventilation could be explained by a weakness of the respiratory muscles or a reduction of the central respiratory drive. The functional disturbances could be diagnosed by spiroergometry without expensive or invasive methods. The findings should be examined more specifically with regard to therapeutic consequences. PMID- 19219769 TI - [Thoracic actinomycosis: diagnostic pitfalls and therapeutic considerations]. AB - We report two patients admitted to our hospital suspected to suffer from cancer in the lung or mediastinum, respectively. Both patients had a diagnosis of thoracic actinomycosis. A 76 year old man revealed pulmonary and endobronchial actinomycosis associated with broncholithiasis. Diagnosis was achieved by bronchoscopy. Therapy with ampicillin/sulbactam was successful. A 36 year old patient presented with bilateral pleural effusions, extended pericardial, mediastinal and pulmonary actinomycosis with pericarditis constrictiva and superior vena cava syndrome. Diagnosis was finally made by cardiac surgery with therapeutic pericardectomy. Prolonged therapy with ampicillin/sulbactam was administered with satisfactory result. Here we discuss the importance to include actinomycosis in the differential diagnosis of pulmonary affections and mediastinal masses in order to avoid diagnostic errors and to limit invasive procedures to the necessary amount. We illustrate the need of an individualized treatment approach. PMID- 19219770 TI - [Baby abandonment and biobanks--with what does the German Ethics Council have to deal?]. PMID- 19219771 TI - Acetabular centre-edge angles revisited: applications and limitations in patients with acetabular dysplasia undergoing periacetabular osteotomy. AB - This study evaluates the application and limitations of the acetabular centre edge angles as described by Wiberg (LCE) and Lequesne (VCA) in a group of adult patients with acetabular dysplasia that were treated with periacetabular osteotomy. Fifty hips in patients with a mean age of 30 years (range, 17-45) were identified and a number of radiographic indices were compared pre and post osteotomy. The potential for measurement variation in both the LCE and VCA angle was evaluated and relationships between the centre-edge angles and other radiographic indices were determined. While all hips displayed some degree of lateral deficiency only 19 (40%) of these cases displayed a ''classic'' lateral and anterior deficiency while 12 (20%) were in fact retroverted. The mean VCA in hips with primarily anterior and lateral deficiency (-6.712.5) was significantly lower (p<0.01) than those with uniform deficiency (5.18.3) or those with retroverted acetabuli (8.913.3). Overall the mean VCA angle of 2.3 (SD12.7) and LCE angle of 3.4 (SD9.3) was corrected to 25.8 (SD11.6) and 28.6 (SD8.7) following osteotomy. The VCA and LCE angles were not correlated (r=0.35) and the LCE angle showed no significant correlation to other lateral coverage indices (Tnnis, Sharp). No correlation was seen either in the post osteotomy values, or in the absolute degree of correction. An alternate VCA (aVCA), identifying the most anterior aspect of the acetabular margin as the reference point, was significantly larger (p<0.001) but did correlate (r=0.77) with the traditional VCA. Potential sources of error in measurement were identified and are reviewed. PMID- 19219772 TI - The influence of implant design on periprosthetic bone remodelling of two types of uncemented HA-coated hip stems. A two-year follow-up study using DEXA. AB - Proximal bone resorption and an increased fracture rate in the ABG-I stem has been shown. For these reasons the ABG-I stem design was changed to the ABG-II. In this study periprosthetic bone loss around the ABG-I vs ABG-II is compared to verify if the design changes resulted in improved proximal bone preservation. METHODS: 51 patients were randomised to either the ABG-I or ABG-II hip prosthesis. Periprosthetic BMD change at various time points was measured using DEXA. Between the two groups (age, gender, weight etc.) no statistical difference was encountered. Compared to the baseline at two years the ABG-II preserved bone better proximally (e.g. zone 7: ABG-II: -3.7%, ABG-I: -11.9%, p=0.05) than the ABG-I. Distally, the trend was opposite and less bone loss was measured for the ABG-I than the ABG-II in zones 3, 4 and 5 (n.s.). CONCLUSION: this study confirms the philosophy behind the design changes from the ABG-I to ABG-II stem where increased elasticity, more proximal HA-coating, a shorter and distally polished stem, were meant to reduce proximal bone resorption. In future this may lead to fewer periprosthetic fractures and to less complicated revision surgery. PMID- 19219773 TI - Distal femoral cortical hypertrophy: predisposing factors and their effect on clinical outcome. AB - The clinical and radiographic features of 109 hybrid total hip replacements performed between 1986 and 1992 in 96 consecutive patients were retrospectively reviewed. A cementless Harris-Galante (HGP1) cup and a 32 mm monobloc straight Muller stem were used in all cases. The overall prevalence of distal femoral cortical hypertrophy (DCH) was 43.1%. Male sex, underlying diagnosis of avascular necrosis and Charnley grade A were among the factors that predisposed to the development of DCH. The prevalence of stem radiolucencies was lower in the group of patients who developed DCH compared with those without hypertrophy. The Harris Hip Score ranged between 5 and 54 preoperatively and the average HHS was more than 80 postoperatively. No association was found between DCH and thigh pain. PMID- 19219774 TI - Long-term results of an uncemented straight femoral shaft prosthesis. AB - The first consecutive series of 250 implantations of an uncemented, proximally anchored straight femoral shaft prosthesis in 236 patients is included in this prospective follow-up study. Average time of follow-up evaluation was 11.8 years (range 10.6-13.7 years). At follow-up, 41 patients with 44 hips had died and three could not be located. Eight hips were revised, two for infection, one for recurrent dislocation, two for rapid subsidence due to component undersizing and one for aseptic loosening of a varus-malaligned stem; two radiologically well fixed stems were revised during acetabular revision. Survival estimate showed an overall survival of 96.6% after 14 years (confidence limits: 98.4% (upper) and 92.8% (lower)). The median Harris hip score at time of follow-up was 81.4 points; clinical results were compromised by high loosening rates (68.8%) of the threaded cup, which had been used in 165 of the 250 cases. Radiologically, tiny reactive lines (<2 mm) were frequently present in the distal zones of the femoral shaft, but could be found in the proximal anchoring zone in only one case. In conclusion, the long-term follow-up results with this femoral component are encouraging and are comparable to those of modern techniques of cementing in primary total hip arthroplasty. PMID- 19219775 TI - Limb length restoration during total hip arthroplasty: use of a caliper to control femoral component insertion and accurate acetabular placement relative to the transverse acetabular ligament. AB - Current methods for restoring or preserving limb length following total hip arthroplasty largely depend on restoring the distance between a fixed point on the pelvis and femur. Each of these techniques allows length correction to be made by combining the effects of both acetabular and femoral height into a single measurement. These methods help to minimise inequality but are anatomically flawed, as they do not allow independent control of placement of the femoral and acetabular components which both contribute to leg length. To address this we present and evaluate a technique that uses a caliper to control the vertical placement of the femoral component and the transverse acetabular ligament to control the vertical height of the acetabular component. Limb lengths were measured in 200 patients who had undergone primary total hip arthroplasty using this technique. Using this method 94% had a postoperative limb length inequality that was 6mm or less (average, +0.38 mm). The maximum measured limb length inequality was +/-8 mm. PMID- 19219776 TI - A radiological and laboratory assessment of polyethylene wear in failed total hip replacement. AB - This study assessed the reliability of radiological assessment of polyethylene wear in loose total hip replacements (THRs) by comparing it with a laboratory assessment using the shadowgraph technique. It was performed in Avon Orthopaedic Centre, Southmead Hospital, from September 1997 to June 1998. True linear wear depth measured using the shadowgraph technique was 42% greater than that determined from the radiological femoral head eccentricity in standard non weightbearing plain x-rays. There was an inverse statistically significant correlation between cup inclination angle and the volumetric wear rate. The linear wear rate, as determined with the shadowgraph technique, was greater in retrieved cups during the first six years postoperatively, but reduced to a lower level in cups retrieved after that time of service. PMID- 19219777 TI - Metal-on-metal joint arthroplasty: potential lifetime longevity. AB - We have studied a historical group of patients who underwent metal-on-metal hip arthroplasty more than 30 years ago. Those retaining their original metal-on metal articulation (average follow-up 33 years) had an average Harris Hip Score of 75 (range 53-93) and an average Musculoskeletal Tumour Outcome Score of 75 (range 63-90). Those that have been revised (at an average of 22 years) maintain good functional outcome. The original implant is still in situ in five of the 11 survivors (follow-up 26-38 years). Retrieval analysis using a Mitutoya form tracer and an electron microscope demonstrated the self-polishing capacity of the metal-on-metal articulation. The surface roughness (Ra) of the worn articulating surface (Ra 0.05 mm) was smoother than that of the original unworn surface (Ra 0.32 mm). We believe our case series highlights the potential longevity of metal on-metal articulation. PMID- 19219778 TI - Fatal pulmonary embolism following hip and knee replacement. A study of 2153 cases using routine mechanical prophylaxis and selective chemoprophylaxis. AB - In order to prevent fatal pulmonary embolism, TED stockings, foot pumps and early mobilisation on the second post-operative day are used at our centre. Only patients deemed to be high risk (previous DVT/PE or obese) are given clexane as inpatients and warfarin for six weeks post-op. From the hospital database 1137 primary total hip replacements and 1017 primary total knee re-placements were identified and the figures were confirmed with the theatre implant order books. The cause of death for those patients on the database, now deceased, was obtained from the coroner. Where a postmortem had not been performed the patient was assumed to have died of a PE. Within three months of surgery, a fatal PE rate of 0.09% (95% CI 0.00-0.26%) following hip replacement and 0.20% (95% CI 0.00-0.46%) after knee replacement was found. Thirty-four patients had been discharged on warfarin according to the pharmacy records. We would therefore not recommend the routine use of chemical thromboprophylaxis following joint replacement. PMID- 19219779 TI - Previous medical problems in 326 consecutive hip fracture patients. AB - Pre-existing medical problems of elderly patients with hip fracture are seldom considered in orthopaedic literature, although they are indisputably the most important determinants for mortality, morbidity and final outcome. It is the purpose of this study to determine these problems in our hip fracture patients. Previous medical disorders and treatments, age, sex and type of fracture were prospectively recorded from all patients over 65 years old, diagnosed with hip fracture in a tertiary university general hospital during 2004. There were 326 patients who fractured their hip (81.04 hip fractures/100,000 people/year) (83.67.3 years old) (85.3% female). The patients existing medical conditions included hypertension (53% of patients), diabetes (19%), dementia (18%), cerebrovascular disease (11%), cataracts and/or blindness (10%), cardiac arrhythmia (9%), chronic obstructive pulmonary disease (9%), heart failure (8%), ischaemic heart disease (7%), psychiatric disorders other than dementia (7%), peptic ulcer (7%), and Parkinson's disease (5%); only 7% had no known significant medical problem beyond their fracture. Cardiovascular and neurological disorders, the most frequent, were also the most dangerous as potential sources for complications and difficulties during anaesthesia, surgery, immediate postoperative period and rehabilitation. Diabetes, the second most frequent diagnosis, complicated any other existing condition. PMID- 19219780 TI - Knitted outer gloves in primary hip and knee arthroplasty. AB - A randomised trial was carried out to determine the rate of perforation to inner gloves when comparing latex with knitted gloves during hip and knee arthroplasty. Members of the surgical team were randomised to wear either two pairs of latex gloves (standard double gloving) or a knitted glove on top of a latex glove. In addition, participants completed a visual analogue assessment of their overall satisfaction with the gloves. A total of 406 inner gloves were tested for perforations over a four-month period: 23% of inner gloves were perforated when latex outer gloves were used and 6% of inner gloves were perforated when knitted outer gloves were used. In total, there were 64 perforations to the inner gloves; only one of these perforations was detected by the glove wearer. Wearing knitted outer gloves during hip and knee arthroplasty statistically significantly reduces the risk of perforation to inner latex gloves (p<0.0001). PMID- 19219781 TI - Extracting the tip of a fractured DHS wire from the hip joint. AB - Dynamic Hip Screw fixation is a very commonly performed operation, but one not without complications (1-3). To stabilise the fixation the guide wire can be advanced into the subchondral bone. An unusual complication of this, and one not previously documented in the literature, is that the tip of the guide wire can fracture. We describe a simple method of removing the tip of the guide wire. PMID- 19219782 TI - Anterior or posterior: does the surgical approach to the hip influence the quality of the femoral cement mantle? AB - The purpose of this study was to determine whether the surgical approach to the femoral neck during total hip arthroplasty affects the quality of femoral cement mantle in terms of thickness and defects, both of which have been implicated in the development of aseptic loosening. A retrospective radiological review of 50 Stanmore Hip femoral stems inserted using the anterior approach and 50 inserted via the posterior approach was performed, with analysis by zone of cement mantle thickness and number of defects. This study demonstrates that the anterior approach is associated with an increase in the frequency of deficiencies in the cement mantle. We recommend that surgeons using the anterior approach are wary of this increased risk and adopt measures, such as the use of distal centralisers, to minimise its occurrence. PMID- 19219783 TI - Long-term clinical and radiographic outcome of the PPF system in ceramic-on polyethylene hip bearings. AB - From 1990 to 1991 we implanted 176 non-cemented proximal press-fit (PPF) total hip arthroplasties (THA) with ceramic-on-polyethylene articulation in 170 patients. Of these, 119 patients (122 THA) were followed from 104 to 129 months. Four cups have been revised for aseptic loosening, but revealed no radiolucencies of the stem. Two infections and two dislocations occurred. The median postoperative Harris hip score was 91. Ninety-eight per cent of the patients were satisfied with the outcome, 83% were free of pain and 67% walked without a limp. Serial radiographs showed stable fixation with bone ongrowth in nearly all arthroplasties except for four cups. Stem radiolucencies were first seen within 12 months, but were of no clinical relevance. Proximal periprosthetic bone resorption of the stem was seen in 18%. Radiolucencies occurred because of polyethylene debris-induced granulomas. The PPF system yields satisfactory long term results in patients with primary and secondary hip osteoarthritis and dysplasia. PMID- 19219784 TI - The distribution of cement penetration into bovine acetabular bone utilising various pressurisation techniques. AB - Most mechanical failures of acetabular fixation within ten years of primary cemented hip replacement are attributable to a failure to achieve good initial fixation of bone and cement. Several studies have suggested that to achieve good fixation between bone and cement between 3 mm and 5 mm of cement penetration into bone is desirable. Whilst several acetabular cement pressurisation systems exist, little is known about the effects of cement pressurisation on the pattern of penetration of cement into cancellous bone within the pelvis. The current paper assesses the penetration of bone cement into bovine acetabular bone by various pressurisation techniques. We found that pressurisation with the use of a swab in glove or the DePuy pressuriser led to improvement in cement penetration both in terms of maximum depth and also percentage cover of the cup by cement (p<0.05). There was no significant difference in mean maximal penetration between the use of the pressuriser and the swab in a glove technique (p=0.3). If only a plain cup was used as a pressuriser, the penetration achieved was significantly lower in comparison to use of a pressuriser (p<0.02). When compared to previously documented acrylic models, the pattern of intrusion noted in the bovine model was different: penetration was lower at the equatorial and base regions of the acetabulum. PMID- 19219785 TI - Early diagnosis of delayed vascular injury associated with revision total hip arthroplasty. AB - We describe a case report where the acetabular prosthesis migrated medially into the pelvis over a period of four years, following revision hip arthroplasty. This was an acute on chronic progressive displacement that caused a slow bleed from the left external iliac artery. Contrast computed tomography is the investigation of choice to diagnose such an injury and demonstrate the slow bleed as opposed to digital subtraction angiography (DSA) arteriogram. PMID- 19219789 TI - Triple pelvic osteotomy for the management of painful developmental hip dysplasia in the young adult. AB - We present a consecutive series of 24 adolescent and young adult patients who have undergone a triple pelvic osteotomy as treatment for symptomatic developmental dysplasia of the hip. With a mean duration of follow-up of 6.5 years (range 2 -16 years) we report good improvements with regard to pain and function in 22 of the 24 patients. Radiological improvements were consistently demonstrated in the centre edge angle and the acetabular index. There were no major complications. When dealing with this difficult problem in young patients we maintain that a redirectional pelvic osteotomy is an appropriate means of treatment in symptomatic patients with a congruent joint. PMID- 19219790 TI - Evaluation of the learning curves associated with uncemented primary total hip arthroplasty depending on the experience of the surgeon. AB - The aim of the paper is to prove and to portray the learning curve in total hip arthroplasty. This prospective study included 168 patients who were operated on by three surgeons, all demonstrating different degrees of experience in performing total hip arthroplasty. Perioperative complications and postoperative radiographs were analysed. Patients were re-examined clinically and radiologically five years postoperatively. In addition, a second study with 41 patients was performed, evaluating especially the learning curve of the highly skilled surgeon in using a femoral neck prosthesis. Radiological complications presented by the first 84 operated patients (2 nd study: 25 hips) were significantly higher than those in the following 84 patients (2 nd study: 26 hips). The learning curve of all surgeons was completed after 20 operations regardless of their experience. Clinical results at follow-up were not influenced by the learning curve. Intensive preoperative planning and exchange of experiences will reduce the length of the learning curve within and outside every clinic. PMID- 19219791 TI - Anterior capsular repair in total hip arthroplasty performed for late-presenting displaced femoral neck fractures. AB - Dislocation after total hip arthroplasty performed for femoral neck fractures occurs frequently due to the significantly higher range of movement of the postoperative hip joint. Proper wound closure can prevent the loss of structural support provided by the capsule and myofascial structures, and is essential for postoperative hip stability. On 32 consecutive patients (four men, 28 women; mean age 64 years, range 48-83 years) with unilateral late-presenting displaced femoral neck fractures, a careful anterior capsule repair technique during total hip arthroplasty was performed by a single surgeon between 1997 and 2000. At one year follow-up, no dislocation had occurred in any patient. Intraoperative protection and careful anatomic reattachment of the hip joint capsule may increase the stability of the hip joint after total hip arthroplasty and lead to a lower incidence of dislocation. PMID- 19219792 TI - Greater trochanteric pain after total hip arthroplasty: the incidence, clinical outcome and associated factors. AB - We evaluated any residual pain around the hip joint 11 to 23 months post-surgery in 172 consecutive patients who underwent total hip arthroplasty (THA) for primary or secondary osteoarthritis during 2002 at Sundsvall Hospital, Sweden. Patients with suspected greater trochanteric pain (GTP) were matched with controls from the same cohort. The two groups were assessed with the Western Ontario and McMaster Universities Arthrosis Index (WOMAC) and were examined for localised tenderness over the trochanteric area by algometer. The pre- and postoperative radiographs including femoral offset were evaluated. We found that 21 patients (12%) had GTP with a male to female ratio of 1:6. The WOMAC index revealed a reduction in the clinical outcome in this group. An associated factor of significance was postoperative uncorrected lengthening of the operated limb equal to or more than one centimetre. We found no correlation between the prevalence of GTP and body weight or length, femoral offset or size of the prosthetic components used. PMID- 19219793 TI - Thirty-two millimetre heads and accelerated polyethylene wear in total hip arthroplasty. AB - In total hip arthroplasty 32 mm heads are often used as the greater diameter is thought to contribute to improved stability. However, greater head diameter can also lead to greater volumetric wear. We compared polyethylene wear between patients with 32 mm heads and those with smaller heads. All patients who had Harris Galante1 metal-backed, uncemented cups inserted between 1986 and 1996 at our institute were annually reviewed clinically and radiographically. Accelerated polyethylene wear was noted between the fifth and seventh postoperative year in 17 out of 165 total hip replacements with 32 mm heads, but only in 21 out of 961 total hip arthroplasties with smaller femoral heads. In all 17 THRs, accelerated wear was also associated with thin polyethylene (<6 mm). This paper highlights a potentially serious problem. When 32 mm metal heads are used with uncemented cups and polyethylene liners, there is a risk of accelerated acetabular wear in patients with thin polyethylene. PMID- 19219794 TI - Distal femoral stem tip resection for thigh pain complicating uncemented total hip arthroplasty. Five patients followed up for 6-10 years. AB - Thigh pain after total hip arthroplasty can be a significant management problem both to patient and surgeon. In many cases, the pain is caused by femoral stem impingement due to its varus position. We present the technique of resection of a segment of distal femoral stem through a minimal incision as treatment of this problem. Five patients with a follow-up of 6 to 10 years are discussed. In all patients thigh pain was explained by the radiographic appearance of the varus stem position and reactive bone formation around the tip of the stem. A bone window was created and resection of the tip of the stem was performed. Four of the patients completely recovered after surgery and one patient, with Pagets disease, required additional interventions due to stress fracture. This is a very simple and effective method of treatment of thigh pain due to femoral stem impingement in healthy bone. PMID- 19219795 TI - Acetabular dome retroversion: radiological appearance, incidence and relevance. AB - Interest in acetabular version arose from the study of unstable developmental dysplastic hips (DDH). Initial studies and clinical observations described the dysplastic hip as being excessively anteverted. Doubts on this view arose from analysis of complications such as persistent posterior subluxation after acetabular reorienting procedures. Computed tomography fails to determine conclusively whether or not the dysplastic acetabulum is abnormally anteverted. Controversy evolves from different methods of measuring and from the fact that the acetabular opening gradually spirals from mild anteversion proximally to increasing anteversion distally. This renders the measurement of version dependent on pelvic inclination and the level of the transverse CT scan slice. On an orthograde pelvic radiograph, both pelvic inclination and rotation can be controlled. Therefore, acetabular version is best estimated based on the relationship of the anterior and posterior acetabular rims to each other on an orthograde pelvic radiograph. Acetabular retroversion has been found to be a characteristic feature of specific hip disorders such as post-traumatic dysplasia, proximal femoral focal deficiency and bladder exstrophy. In addition, acetabular retroversion has been described in DDH as well as in dysplastic hips in the context of neuromuscular and genetic disorders. Iatrogenic acetabular retroversion can also result from corrective pelvic osteotomies in childhood. Finally, retroverted acetabula may be found in otherwise nondysplastic hips. The relevance of acetabular retroversion is twofold: First, it demands a more individualized approach to acetabular dysplasia because the presence of retroversion will affect the manner in which the corrective osteotomy should be done. Second, the long-term effect of acetabular retroversion is harmful. PMID- 19219796 TI - In vitro monitoring of hoop strains during impaction grafting of the femur. AB - A test was designed to monitor hoop strains around the proximal femur during impaction grafting. Four electrical resistance strain gauges were attached to Sawbone femurs just below the level of the lesser trochanter. Experimentation with these femoral models included the use of a device called an "impactometer", which delivered controllable impaction energies. Test material used was ovine morsellised bone graft prepared from sheep femoral condyles. Instruments used were "X-change" revision instruments from the Exeter hip system. Maximum strains occurred at the medial and lateral cortices and there were lesser strains on anterior and posterior cortices. These findings raise questions about the quality of impaction anterior and posterior to the femoral stem. At the medial and lateral gauges significant differences in levels of strain were recorded at the three different levels of impaction. An individual hoop strain was found to have three components: peak, rapid drop and slow drop. The strain level did not drop back to the starting point at the end of the drop phase but dropped only halfway back to the starting level. This resulted in a rise in strain levels in a stepladder pattern. Strains continued to rise significantly when the impaction process was continued beyond the endpoint of impaction. Early recognition of the endpoint of impaction can save the weak femoral bone from unnecessary high levels of hoop strain. PMID- 19219797 TI - Case report: overnight hospital stay for minimally invasive bilateral total hip arthroplasty. AB - Bilateral total hip arthroplasty was performed on a 58-year-old man. He was successfully discharged home from hospital the following morning, 23 hours post surgery. A direct anterior, minimally invasive approach was used which avoided detachment of hip musculature. Preoperative assessment was carried out, with early multidisciplinary team input. Modified anaesthetic techniques with an ambulatory pain pump were employed. Follow-up in the community was carried out by an outreach team. All these factors were important in a successful early supported discharge. We believe this to be the first such case reported. PMID- 19219798 TI - Psoas abscess associated with infected total hip arthroplasty: a case report. AB - Psoas abscess (PA) is an uncommon disease and its diagnosis is difficult. It can be primary or secondary. Primary abscesses are of unknown origin and are presumably caused by haematogenous or lymphatic spread from a distant infectious focus. Secondary PA is caused by spreading from a contiguous infected structure, such as vertebrae (espondilodiscitis) or mesenteric abscesses (Crohns disease). PA infrequently has been associated with an infection of total hip arthroplasty (ITHA). The correct diagnosis in these cases is difficult due to the clinical similarities between PA and ITHA. Since connection between PA and ITHA is established through the acetabulum, we consider that computed tomography (CT) is the most accurate radiological test because of its efficacy in evaluating the bone structure, and the optimum therapeutic strategy is two-stage replacement surgery. We report one case of PA associated with ITHA and a review of the previous literature. PMID- 19219799 TI - Improved retraction in minimally invasive total hip replacement. PMID- 19219800 TI - The shortcomings of computer-aided measurement of migration for the prediction of failure of three forms of acetabular fixation by survival analysis and migration study to ten years. AB - We studied, to ten years, migration by vertical displacement and angular change using a digitizer and standard radiographs and survival of three methods of acetabular fixation in primary hip replacement. Two implants were uncemented metal-backed components, one version of which was hydroxyapatite-coated. Both types had 28 mm polyethylene inserts. The third type was a cemented all polyethylene cup with a 28 mm internal diameter. All cups articulated with a Freeman neck-retaining stem. The purpose was to see if the early migration data predicted the long-term outcome and could be used to forecast implants that would fail for the purpose of pre-market assessment. No statistical differences were found between the groups in the parameters studied. At the threshold accuracies of our migration measurements (three millimetres and three degrees) at three years, we could not identify components that would subsequently fail. However, migration less than these threshold values and the absence of radiolucent lines were strongly associated with implant survival at ten years. Our system was insufficiently accurate for pre-market surveillance.; PMID- 19219801 TI - A new technique to improve cup inclination accuracy in primary total hip arthroplasty. AB - Accurate inclination and anteversion of the acetabular component is paramount to achieve a stable total hip arthroplasty, prevent prosthetic dislocation and minimize long- term wear. We present and evaluate a simple new technique to improve consistency in cup inclination during primary total hip arthroplasty, based on information available from the preoperative plan. The technique consists of assessing the distance and relationship between the inferomedial border of the acetabular cup and the inferomedial margin of the teardrop, with the use of a measuring probe. This simple surgical gesture improved consistency in cup inclination during total hip arthroplasty.; PMID- 19219802 TI - Total hip replacement using hemi-circumferential interposition acetabuloplasty for acetabular deficiency in post-Perthes deformities: technique and long-term results. AB - We describe a technique of hemi-circumferential interposition grafting that allows placement of the cup in the anatomical position of the original acetabulum in the rare cases of post-Perthes or Perthes-like deformities of the femoral head combined with a steep and shallow acetabulum. This technique was performed on 10 hips (9 patients, with an average age of 56 years). One revision was performed for septic loosening after 2.1 years; however, the graft was well integrated and provided sufficient support for the revision cup. All the others had a good clinical and radiological result after an average of 11.4 years with an average Harris Hip Score of 97. The authors have found this an easy-to-use method, which can give an excellent, long-lasting result. It also provides important bone stock for possible subsequent revisions.; PMID- 19219803 TI - Revision hip surgery using the CLS expansion shell. AB - The aim of our study is to assess the outcomes using the uncemented CLS expansion shell in revision hip surgery for aseptic loosening of the acetabular component. Between 1991 and 2000 we used the CLS expansion shell in revision hip surgery in 215 patients (230 hips). We were able to evaluate 196 patients (211 hips) with failed acetabular components. The male/female ratio was 82/114 and the mean age of the patients was 63 years (range 38-71). The main reason for revision was aseptic loosening of the PE cup (184 patients). There were 79 hips with Paprosky defect of the acetabulum type I, 62 hips with type 2A, 57 hips with type 2B and 13 hips with type 2C in the study. The mean follow-up was 8.5 years (range 5-14). Plain radiographs using an AP view of the pelvis and AP and lateral views of the affected hip were taken immediately after surgery and at the latest follow-up. Clinical outcomes were determined using the Merle dAubigne' and Postel score and the Harris Hip score. The mean preoperative Merle dAubigne' score was 7.8 points (range 6.9 to 9.9); at the latest follow-up it was 14.9 points (range: 12.5 to 15.9). The mean preoperative Harris Hip score was 38.6 points (range: 32 to 59); at the latest follow-up it was 82.5 points (range: 38 to 95). Osteointegration of the CLS expansion shell was present in 149 hips (70.6%) at the latest follow-up. There were 31 complications (15%) requiring a second revision. Clinical survivorship of the CLS expansion shell was 90% at five years and 84% at 8.5 years after revision surgery. Radiological survivorship was 78% at five years and 70% at 8.5 years after revision surgery. The CLS expansion shell offers a viable alternative for revision hip surgery with lower bone quality, insufficient bone at the bottom and cavitary defects of the acetabulum.; PMID- 19219804 TI - The quality of web-based information on hip resurfacing arthroplasty: a cross sectional survey. AB - The Internet has become an important source of information for patients. Websites offering medical information are not subject to peer-review. The aim of this study was to evaluate the quality of information available on the web regarding hip resurfacing arthroplasty. The search term hip resurfacing was entered into five commonly used general search engines. The 20 highest-ranked websites from each engine were reviewed, after eliminating duplicate and non-relevant sites. Each site was then given a quality score (maximum of 20) according to predefined general and specific criteria. Our search identified 40 unique websites. Commercial websites were the most common (n=17). Quality scores ranged from 8 to 18, indicating that the quality of information on the web varied widely. The mean score for all sites was 11.7, SD 2.6, and only four websites scored >15. Fifteen sites did not mention the disadvantages or complications specific to hip resurfacing. The quality of information on the Internet regarding hip resurfacing is variable. Only four sites out of 40 were deemed to be of high quality. Orthopaedic surgeons need to be aware of these limitations, and take the lead in educating and counselling their patients.; PMID- 19219805 TI - Evaluation and treatment of young adults with femoro-acetabular impingement secondary to Perthes' disease. AB - Hip pain and loss of motion in young adults with previous Legg-Calve-Perthes Disease may be caused by anterior femoro-acetabular impingement. Eleven patients (12 hips) with the chief complaint of groin pain and significant proximal femoral deformity were treated. Gadolinium-enhanced magnetic resonance arthrography in ten patients indicated labral injury and adjacent acetabular cartilage lesions in nine hips. A surgical dislocation of each hip confirmed that there was impingement induced intra-articular injury consistent with the pathology indicated on the MRI. Reshaping of the femoral head, with correction of the femoral head/neck offset, and treatment of the acetabular rim pathology was performed for each hip in conjunction with other procedures for the proximal femur. Correction of the impingement and increased range of motion could be visualized intra-operatively. At a mean follow-up of 33 months, half of all patients were pain-free and all had improvement in pain compared with preoperatively. Ten patients had an improved range of motion and two a slight decrease. No additional necrosis following the dislocation of the femoral head was seen.; PMID- 19219806 TI - Os acetabuli in femoro-acetabular impingement: stress fracture or unfused secondary ossification centre of the acetabular rim? AB - Ossicles located at the acetabular rim are generally referred to as unfused secondary ossification centres and are named "os acetabuli". They are also observed in severely dysplastic hips, where they are considered as fatigue fractures of the acetabular rim due to overload. In a retrospective study, we evaluated the radiographs of 495 patients, who were treated surgically for femoro acetabular impingement. In 18 hips (15 patients) a large osseous fragment at the anterolateral acetabular rim was found. All patients presented radiographically with a femoral head showing an aspherical extension producing a "cam" impingement. Sixteen hips had a retroverted acetabulum, indicating anterior overcover. Preoperative MRIs available in 12 patients showed the presence of a fragment composed of labrum, articular cartilage and bone. The gap between the stable acetabulum and the rim fragment had a vertical orientation. All patients had been exposed to a physically demanding profession or contact sport and in 15 hips no memorable traumatic episode was present. The mechanism leading to this acetabular rim fragment is thought to be fatiguing due to femoro-acetabular impingement. The aspheric portion of the head is jammed into the acetabulum and with time causes a stress fracture of the retroverted portion of the acetabulum. True "Os acetabuli" are morphologically similar, but the orientation of the cartilaginous growth plate is more parallel to the joint surface, in contrast to the hips in the current study, where the separation line was perpendicular to the joint surface. The clinical importance is that the presence of an acetabular rim fragment in the non-dysplastic hip must raise suspicion of femoro-acetabular impingement.; PMID- 19219807 TI - Outcome of bilateral hip hemiarthroplasty for displaced fracture of the hip. AB - Our aim was to evaluate the outcome in patients with bilateral non-simultaneous hip fracture treated with hemiarthroplasty at our institution between 1997 and 2003. We looked for any important modification factor that could change the devastating functional and social results. Twenty-two patients with a mean age of 79 at the initial fracture and 82 at the subsequent fracture were evaluated; all were Garden grade III-IV. The second fracture happened a mean of 28 months after the first one. The outcome measurement was the incidence of hip pain, recovery of pre-injury level of ambulation and activities of daily living at a minimum follow up of one year. Following the first hemiarthroplasty patients deteriorated in their walking capacity and other activities of daily life, and this happened to an even greater extent after the second operation. With respect to walking ability (measured in metres) we concluded that it improved if a modular hemiarthroplasty was used after both fractures (p<0.05) and that worse functional outcomes were observed when different kinds of hemiarthroplasties were used in the same patient.; PMID- 19219808 TI - The trochanteric gamma nail versus the dynamic hip screw: a prospective randomised study. One-year follow-up of 146 intertrochanteric fractures. AB - In a prospective, randomized trial we compared the trochanteric gamma nail (TGN) and the dynamic hip screw (DHS) in the treatment of 146 intertrochanteric fractures. Follow-up was after four and 12 months. The operation time was significantly shorter in the DHS group. At discharge the need for walking aids was less in the DHS group. There were no differences in intraoperative blood loss, medical complications, mortality or length of hospital stay. Major fracture complications occurred twiceas often in the TGN group compared with the DHS group, however they were not statistically significant. Any potential for the TGN leading to a less invasive procedure and a more rapid postoperative mobilisation could not be demonstrated. Compared with the TGN we prefer the DHS for most intertrochanteric fractures in a setting where the majority of these fractures are treated by younger doctors and not by highly specialized hip/trauma surgeons. The TGN may have advantages in selected intertrochanteric fractures.; PMID- 19219809 TI - Dissociation and implantation of Exeter femoral instrumentation during total hip arthroplasty. AB - Proprietary femoral plug introducers are routinely used to insert intramedullary plugs during total hip arthroplasty. The cement restrictor pin from an Exeter intramedullary plug introducer may become detached during insertion of the cement restrictor plug, or during sponge packing of the femur. We suggest some precautions that should be taken to prevent this from occurring and advise that the patient is unlikely to experience any long-term adverse reactions to the presence of the restrictor pin in the intramedullary canal.; PMID- 19219810 TI - Intrapelvic protrusion of a dynamic hip screw. AB - The sliding hip screw, sometimes called the dynamic hip screw (DHS) has been a successful method for treating hip fractures. Nevertheless, mechanical complications of this system have been reported from time to time. They are mainly the result of instability of the fracture, bone quality and technical errors occurring during surgery. We report a case of an intrapelvic protrusion of a sliding screw after a basicervical fracture was treated with a DHS and a 6.5 mm cannulated screw, and its surgical solution using a hybrid total hip arthroplasty. In our case we think the mechanical failure was due mainly to technical errors during surgery. Despite that, we believe the sliding hip screw is the implant of choice for basicervical fractures. Using an additional K-wire during the technique is sufficient to avoid rotation of the proximal fragment and no additional fixation is necessary.; PMID- 19219811 TI - Technical tip: a useful aid for the preparation of morsellised bone graft. PMID- 19219813 TI - Foreword. AB - Excellent long-term results have been achieved with cemented implants. However, the profile of the patient presenting for total hip replacement has changed. Pain and disability are no longer the only indications for surgery. The millennium patient, em-powered with information, seeks surgical intervention to restore quality of life. Thus, ever younger cohorts of patients are presenting for total hip replacement. Conservative surgery embraces both soft issue sparing and bone conservation. If the surgeon is confident the hip he is going to use will outlast the patient he should use it. Otherwise he should consider the next operation and conserve bone. Any implant that makes contact with the diaphyseal cortex or is ingrown distally will offload distally with consequent proximal stress protection. Bone conservation is achieved both by taking less bone at the time of surgery and by optimising the physiological loading of the proximal femur to preserve bone in the longer term. This supplement reviews the evolution and the clinical heritage of a short, stemless metaphyseal loading implant the PROXIMA. Elegant biomechanical studies have shown that the PROXIMA is not a 'fit and fill' prosthesis. Excellent axial and rotational stability can be achieved within the cancellous bone, and the implant is suspended in and moves in consort with the surrounding cancellous bone. This reduces the shear stresses at the fixation interface and optimises load transfer in the metaphysis. Bone densitometry demonstrated that the shorter second generation customised Santori stem avoided subtrochanteric buttressing and enhanced the load transfer to the proximal femur with consequent bone remodelling. Using DEXA to compare the periprosthetic bone density of the short stem with a variety of other commercially available cementless femoral implants confirmed that the PROXIMA preferentially loads the metaphyseal bone. The 'round the corner' technique for the insertion of the PROXIMA is ideally suited for minimally invasive surgical approaches. The 'MicroHip' technique uses a section of the anterior Smith-Peterson approach. This is a truly internervous and inter-muscular plane which provides excellent access to the hip joint. Minimal soft tissue disruption, together with bone preservation, makes PROXIMA a truly conservative op-tion for the millennium patient. PMID- 19219814 TI - Early clinical experience with a metaphyseal loading implant: Why have a stem? AB - This is a prospective cohort study of patients undergoing total hip replacement using an uncemented, proximally coated stem with a short tapered distal stem. Patients with a minimum of 24 months of clinical and radiological follow-up were included in this study. There were 18 males and 34 females. Average age at surgery was 40 years. The diagnoses included developmental dysplasia of the hip in 20 patients, osteoarthritis in 15, avascular necrosis of the femoral head in nine, and inflammatory arthritis in three. Average follow-up was 46 months (range 25-78). There was no evidence of subsidence of any stem at the final follow-up. Interface buttressing was observed in 52 patients on the AP view and in 32 on the lateral view. This was seen in Gruen zones 1, 7, 8 and 14. Heterotopic ossification was seen in 15 patients. Distal cortical hypertrophy was seen in three patients. Three patients had one episode of dislocation and two patients had superficial wound infection. None of the hips has been revised and there was no radiological evidence of aseptic loosening. The high percentage of patients exhibiting condensation of the bone and interface buttressing at the lower reaches of the metaphysis suggest the objective of proximal loading is being achieved. We believe that early stability and durable biological fixation can be achieved by this cementless implant, in which the stem is designed to avoid any contact with the diaphyseal cortex. PMID- 19219815 TI - Bone remodelling in THA: A comparative DXA scan study between conventional implants and a new stemless femoral component. A preliminary report. AB - Periprosthetic bone loss is a major cause for concern in patients undergoing total hip arthroplasty (THA). There are many different factors that may determine the pattern of bone loss and bone remodelling following THA, such as the quality of the bone before the hip replacement, skeletal bone mass at the time of the operation, material and method of fixation and implant design. Recent developments in dual-energy X-ray absorptiometry (DXA) have made it possible to quantify bone mineral density (BMD) to evaluate changes around the prosthesis and to measure bone stock and bone density redistribution after a total hip replacement. In this cross-sectional multicentre clinical study the DXA method was used to compare bone mass after uncemented THA of a custom-made stemless design with five groups of conventional cementless implants (Alloclassic, Mayo, CFP, IPS, ABG). The adaptive bone changes of the proximal femur three years after implantation were evaluated. Periprosthetic BMD was measured in 130 subjects in the seven regions of interest (ROI) based on Gruen zones. Significant differences were found between the stemless implant and the other five groups in zones 1, 4 and 7. The CFP, IPS, and ABG groups showed decreased BMD in ROI 1, and the Mayo, IPS and Alloclassic in ROI 7. An increased BMD in ROI 4 was observed in the Mayo, IPS, ABG and Alloclassic groups. The results of the present study suggest that a conservative stemless implant with complete proximal load transfer produces a homogeneous and more physiological redistribution of bone density, allowing maintenance of proximal periprosthetic bone stock. PMID- 19219816 TI - Bone preservation with a conservative metaphyseal loading implant. AB - Proximal load transfer and the absence of distal fixation of the stem are crucial to obtain the best behaviour of the femoral bone after total hip replacement. In this study, dual energy X-ray absorptiometry (DEXA) was employed to understand and compare the bone density changes and thus the re-distribution of mechanical forces in two different extra short stems. Two cohorts of ten patients were included in this retrospective study. Both implants are custom-made and present a well defined lateral flare. The first model is fully coated and presents a short stem (Group A), the second is an unstemmed metaphyseal implant with a polished tip (Group B). DEXA scans were obtained in all patients at the two-year follow up. A higher BMD was detected in ROIs 1, 2, 4, 5 in Group B confirming a preservation of the proximal bone mass and thus indirectly a more proximal load transfer. The results obtained confirm the importance of the geometry of the implant on proximal bone density. Loading both medial and lateral proximal femoral flares and the complete absence of the diaphyseal portion of the stem provide the optimal bone remodelling of the femur after total hip replacement. PMID- 19219817 TI - Biomechanics of a new short-stemmed uncemented hip prosthesis: An in-vitro study in human bone. AB - The migration pattern, cyclic motion, system stiffness and failure load of a new short-stemmed hip prosthesis were compared to a clinically successful shaft prosthesis during the initial loading phase. The influence of implant-sizing on mechanical stability was also investigated for the new stem, in particular with relation to the bone quality. Prostheses were implanted in paired human femora and loaded cyclically up to 3515 cycles. Relative displacements between prosthesis and bone were measured using a 3D-camera and reflective marker system. Migration of the new stem was predominantly into varus and was higher than for the shaft prosthesis. The test set-up was proposed to simulate a worst-case loading scenario since muscle forces, which tend to reduce bone deformation, were not simulated. It could therefore be expected that clinical migration of the implants would be less pronounced than that observed in this study. Cyclic motion for the new stem was similar to that for the clinically successful shaft prosthesis, suggesting that bone ingrowth could be expected for the new stem. No significant difference in fracture load was observed between the implants. The system stiffness with the new stem was lower than for the shaft prosthesis, indicating more physiological load transfer. Smaller implant sizing with 'cancellous' fixation seems favourable, as it led to similar migration and smaller cyclic motion values than with 'cortical' fixation. A trend for higher cyclic motion and migration was observed in femora with poor bone quality. Hence, sufficiently good bone stock is necessary, when implanting the new short-stemmed prosthesis. PMID- 19219818 TI - Ultra-short stems with proximal load transfer: Clinical and radiographic results at five-year follow-up. AB - Bone stock preservation is crucial when performing THR in young patients. Following this philosophy, an original ultra-short custom-made implant with extensive proximal load transfer was developed. It presents three very innovative features: absence of the diaphyseal portion of the stem, a well defined lateral flare with load transfer on the lateral column of the femur and a very high femoral neck cut. These innovations resulted in a very conservative implant both of the bone stock and the soft tissues. In this study we reviewed the X-rays of 111 patients with 131 primary total hip replacements performed with this implant. The average pain score using the Harris Hip Score system, at an average of five years after surgery, was 42 of 44 points; 95 per cent (124) of the patients had no or slight pain. We had no cases of thigh pain. None of the patients required a femoral stem revision. Two cases had to be re-operated for polyethylene liner exchange but the femoral implant was stable and left in place. At five years, all implants appeared radiographically stable with well maintained proximal bone stock. It was concluded that the geometry of this implant provides significant initial stability, which seems to be preserved throughout a long follow-up period. This study validates the assumption that torsional loads can be controlled even without the diaphyseal portion of the stem and that neck preservation combined with lateral flare support guarantees a more natural loading of the femur. PMID- 19219820 TI - The new orthopaedic patient. PMID- 19219819 TI - MicroHip: A minimally invasive procedure for total hip replacement surgery A modified Smith-Petersen approach. AB - In this article we present the surgical technique of our anterior minimally invasive approach to the hip joint. This is aligned along an interneural plane and makes the approach truly minimally invasive. This technique is performed in a lateral decubitus position on a normal operating table; there is no need for a specific orthopaedic or fracture table. As leg traction is not necessary, this enables the approach to be used routinely. Most of the instruments used for this procedure are standard instruments; only the acetabular reamers and positioners are angulated and specifically designed. The angled instruments (MI minimally invasive) are especially useful when performing hip replacement in obese patients. An excellent view of both the acetabulum and the proximal femur can be obtained through an incision of only 6-8cm, using this MI approach. The approach follows the interval between the tensor fascia lata muscle and the rectus muscle using a section of the anterior iliofemoral approach described by Smith-Petersen. No tendon or muscle is cut or detached. The joint capsule is split and left in place. The hip joint is not dislocated and we perform the osteotomy of the femoral neck in situ. To date we have performed over 500 MicroHip operations. We have not observed any nerve lesions or fractures of the trochanter. Our experience to date shows that the method we have de-veloped allows us to operate reliably on virtually any patient. Preliminary results also show that important factors like length of stay, pain, blood loss and return to work can be reduced significantly. The MicroHip technique is being used by more and more different clinics all over the world and can be applied with success after suitable training. PMID- 19219821 TI - The new orthopaedic hip patient. PMID- 19219822 TI - The educated patient in the information age: caveat emptor? PMID- 19219823 TI - Tribology considerations for hip joint articulations in relation to the "new orthopaedic patient". AB - The purpose of this review is to examine alternative bearings used in total hip arthroplasty (THA) and discuss the specific tribologic needs of the "New Orthopaedic Patient". As orthopaedic patients today are younger and more active, there is a clear need for hip joint implants and articulations minimising the amount of wear and guarantying better stability. Recent modern developments in tribology with highly cross-linked polyethylenes and hard-on-hard bearings allow the safe and effective use of larger diameter articulations in THA. PMID- 19219824 TI - Early outcomes following minimally invasive total hip arthroplasty using a two incision approach versus a mini-posterior approach. AB - We evaluated early clinical results and radiographic findings of MIS THA using 2 incsion approach and mini-posterior approach. A consecutive series of 35 patients (40 hips) with 2-incision THA and a consecutive series of 35 patients (36 hips) with mini-posterior THA performed in the same period were evaluated. The patient groups were not randomised. At the mean follow-up of 20.2 months (range, 12-36 months), the 2-incision group had significantly earlier ambulation, driving ability and return to work. However, it had significantly more operative time, blood loss, blood transfusion and complications. Regarding radiographic findings of 2-incision group and mini-posterior group, the mean radiographic acetabular abduction of was 46.7 degrees (89% was within 35- 55) and 44.8 degrees (84% was within 35-55), respectively. The femoral stem was in neutral position in 80% and 83%, respectively. There was no statistical difference of the implant position between groups. Although MIS 2-incison THA provided faster, early ambulation than the mini-posterior THA, surgeons have to weigh the advantages and disadvantages of this technique including, increased operative time, blood loss and their familiarity with similar standard incisions and landmarks. PMID- 19219825 TI - Improved accuracy of acetabular component position and leg lengths with the two incision total hip replacement technique. AB - We sought to determine whether a non-originating surgeon early in his experience with the MIS two-incision technique for THA could place components with appropriate accuracy. Thirty five consecutive hip replacements performed between July of 2003 and March of 2004 using the two-incision technique were closely matched by height and weight with an equal number of control small incision arthroplasties. The two-incision surgeries were performed after the surgeons twentieth case and prior to the surgeons fifty sixth two-incision case. We assessed immediate postoperative films for leg length discrepancy and acetabular position, and compared the two groups by chi squared analysis. Both leg length accuracy and abduction angle were significantly superior with the two-incision technique. No arthroplasty performed with the two-incision technique had a leg length discrepancy of more than 10 mm, versus five in the control group (p<0.025). The abduction of one cup fell outside the 35-49 range in the two incision group, while ten did so in the control group (p<0.01). With this study, we have shown that a non-originating surgeon early in his experience with the MIS two-incision technique can place acetabular components and equalise leg lengths with superior precision. We believe the fluoroscopic control and supine position greatly enhance the surgeons accuracy, and more than compensate for the potentially decreased direct vision with this technique. PMID- 19219826 TI - A modified two-incision minimally invasive total hip arthroplasty: technique and short-term results. AB - Between 2003 and 2005, a total of 425 consecutive cases with a modified two incision minimally invasive total hip arthroplasty (THA) were performed at our hospital. We analysed 225 cas-es with greater than 12 months follow-up clinically and radiographically. The mean operative time was 70 minutes. Patients could walk on crutches at 1.5 days and discontinued crutch use at an average of 3 weeks. Patients were able to walk upstairs without support at 4 weeks. Radiographic analysis showed the mean lateral opening angle and anteversion of the acetabular components were 43.0 and 17.3, and 97% of the femoral stems were in neutral alignment. There was no radiographic evidence of component migration or subsidence at the most recent follow-up. Therefore, from these early results, a modified two-incision THA was found to be an excellent surgical modality, which allows early rehabilitation and does not increase complications when compared to other MIS two-incision THA techniques. PMID- 19219827 TI - Two-incision total hip replacement: Intra-operative fluoroscopy versus imageless navigation for cup placement. AB - The purpose of this study was to describe the surgical technique and to investigate results of a modified two-incision total hip replacement using either intraoperative fluoroscopy or imageless navigation. Twenty-nine patients (30 hips) with a minimum follow-up of one year were enrolled in this study. The patients were evaluated at 3, 6, 12 weeks, 6 months, and 1-year. The functional recovery as represented by the Harris hip score and WOMAC scale were better in the fluoroscopy group of patients at the early postoperative stage (3 wks). Thereafter, both groups showed rapid recovery with no difference in scores. Injury to the lateral femoral cutaneous nerve was the most commonly seen complication and it occurred in 6 hips (fluoroscopy 2; imageless 4). The symptoms were transient and resolved in 6 months in all 6 cases. This study demonstrated that the role of intraoperative fluoroscopy could safely be replaced by an imageless navigation system for the MIS-2 THA. PMID- 19219828 TI - Minimally invasive anterolateral surgical approach for total hip arthroplasty: early clinical results. AB - The purpose of this study is to describe our early experience in the development and use of a minimally invasive, anterolateral approach to the hip joint for total hip arthroplasty (THA). This approach uses a specific intraoperative protocol involving leg positioning to maximise exposure and hip joint visualisation. Between October, 2004 and December, 2004, we operated on 18 male and 29 female patients without selection and in consecutive order. The average patient age at surgery was 66 years (range 28 to 86 yrs) with an average body mass index (BMI) of 26 (range 18 36) and with a majority of patients presenting with osteoarthritis as the primary indication for surgery. The average length of the skin incision was 8 cm and the average incision-to-suture time was 45 minutes. The preoperative average Harris Hip Score was 38 and improved to 70 at 10 days postoperatively, and 92 at three and six months. Similarly, the HHS average pain component score increased from 12.6 preoperatively to 39 at 10 days postoperatively, and 40 at three and six months. An exact neutral implantation of the stem was found in 48% of the patients, between one to three degrees out of neutral in 40% and greater than four degrees of varus in 12%. The average inclination angle of the cup was measured at 44 degrees. No intraoperative or perioperative complications of any kind occurred in this series of 47 consecutive patients. From our experience, the MIS AL approach for THA with patients in the lateral position can be applied without restrictions for patient indication. Improved results can be observed when using the MIS AL approach, especially in the ear-ly postoperative phase. Since the abductors can be completely spared during the surgery, improved long-term results may be expected. Moreover and when necessary, an operative expansion is always possible without difficulties. Overall, the approach yields promising results. PMID- 19219829 TI - The minimal invasive surgery anterior approach with supine patient positioning: a step-wise introduction of technique. AB - The purpose of the review is to report the modified Smith-Peterson technique for use with minimal invasive total hip arthroplasty (MIS THA) with supine patient positioning. The modified Smith-Peterson technique allows for direct visualisation of the acetabulum and is an internerval approach. The MIS anterior approach with supine patient positioning offers a complete intermuscular and internervous access to the hip joint. The use of this technique yields a reduction of muscular trauma, intraoperative bloodloss, and postoperative rehabilitation. The reduction of soft tissue trauma may lead to minimal postoperative pain. While cosmesis is never a reason for using MIS, it does yield great patient satisfaction. The MIS anterior approach with supine patient positioning is a safe and reproducible method, offering an alternative option in MIS THA. PMID- 19219830 TI - Cost-benefit analysis of MIS THA: Model-based analysis of the consequences for Switzerland. AB - The objective of this study was to show model-based economic consequences of minimal invasive surgery total hip arthroplasty (MIS THA). The model is based on hospital and rehabilitation costs and lowering of loss of productivity. This study used conventional THA data from Switzerland and cost-effectiveness MIS THA analysis from the United States. According to our model, a total of 42.1 70.1 million can be saved annually in Switzerland when using MIS THA instead of conventional THA. Annual savings of 7,8 12,9 million result from the significantly shorter hospital stay. The savings potential for rehabilitation costs is 10,5 17,5 million annually. In addition, the productivity loss for the economy is lowered by 23.8 39.7 million annually. According to this economic study we recommend the introduction of MIS techniques in THA. MIS THA surgical techniques may allow the reduction of healthcare costs. Despite the promising economic advantages of MIS THA, clinical studies are necessary to prove long-term benefits. PMID- 19219831 TI - Long-term results with the Wagner metal-on-metal hip resurfacing prosthesis. AB - The purpose of this study was to report the long-term results of the metal-on metal Wagner resurfacing prosthesis. Between January 1991 and August 2004, 54 Wagner resurfacing prostheses were implanted in 52 patients. There were 45 female patients (1 bilateral case) and 7 male patients (1 bilateral case). All the operative procedures were performed using the anterior approach. Of those cases available, the average time to follow-up was nine years. The average Merle dAubign score improved from 8.3 preoperatively to 17.3 postoperatively (p-value < 0.001). Seventeen prostheses required revision. Radiographic evidence of femoral neck narrowing was seen in three prostheses that remained in situ. Combined with a stricter diagnosis and an exact operating technique, the Wagner metal-on-metal surface replacement of the hip joint may offer an alternative surface replacement option. PMID- 19219832 TI - The development of the DuromTM metal-on-metal hip resurfacing. AB - The poor results of conventional total hip replacement in young and active patients have led to a renewed interest in alternative interventions and bearing material combinations. Con-temporary metal-on-metal resurfacing of the hip has emerged as an attractive option. This procedure utilises a low wear producing bearing which minimises the risk of periprosthetic osteolysis, preserves femoral bone stock should subsequent procedures become necessary, and, due to the increased stability and range of motion, allows the rapid resumption of an active lifestyle. We describe the development of the DuromTM metal-on-metal hip resurfacing and report the preliminary results of the first 100 consecutive cases. PMID- 19219833 TI - A prospective randomized clinical trial comparing metal-on-metal total hip arthroplasty and metal-on-metal total hip resurfacing in patients less than 65 years old. AB - The purpose of this prospective randomised study is to compare the early clinical results of the metal-on-metal hip resurfacing to metal-on-metal THA. Two hundred and ten hips were randomised between August 2003 and January 2006 (191 subjects). One hundred and two hips were implanted with an uncemented titanium tapered stem, and an uncemented titanium acetabular component and 28 mm metal-on-metal bearing (THA group) and 103 hips received a hybrid metal-on-metal surface replacement arthroplasty (SRA group). No significant difference was found with the WOMAC or Merle dAubign-Postel scales. However, a significantly higher activity level was found in the SRA group (UCLA score 6.3 versus 7.1, p= 0.037) and a greater percentage of the SRA patients returned to heavy or moderate activities at one - year postoperatively (72% versus 39%, p=0.007). No patient in either group presented with thigh pain one year after surgery. Both techniques present similar complication rates (0.15). This study supports the theory of better functional recovery in the short-term favouring the SRA when compared to THA. The clear benefit of surface replacement arthroplasty over THA is proximal femoral bone preservation. However, the long term survivorship of the SRA will determine the real value of the theoretical advantage. PMID- 19219834 TI - Factors affecting the cement penetration of a hip resurfacing implant: an in vitro study. AB - An understanding of how the cementing technique influences cement penetration is important for surgeons to plan and conduct hip resurfacing procedures. Therefore the aim of this study is to determine the influence of the following parameters on cement penetration: use of pulse lavage, type of cement, and the standing period of the cement. Nine fresh frozen paired whole cadaver femora were used to investigate cement penetration. The femora were divided into three paired groups: (A) compared the use of pulse lavage to no pulse lavage, (B) compared two low viscosity bone cements SULCEM-3 (Zimmer GmbH, Switzerland) and Surgical Simplex (Stryker Orthopaedics, USA), and (C) compared bone cement viscosities using standing times of 1.5 min. and 3 min. The Durom Hip Resurfacing implant (Zimmer GmbH, Switzerland) was used in all groups. A single slice was taken out of the center of each head. Cement penetration ratio (penetration area divided by the bone area enclosed by implant) and mean cement penetration depth were measured. Cement penetration is increased using pulse lavage, and has the tendency to increase with increasing the cement standing time from 1.5 to 3 min. No difference in cement penetration was found when different cement brands of comparable viscosity were used. PMID- 19219835 TI - A strategy for patterning conducting polymers using nanoimprint lithography and isotropic plasma etching. PMID- 19219836 TI - Sequence-specific HCV RNA quantification using the size-dependent nonlinear optical properties of gold nanoparticles. AB - The hepatitis C virus (HCV) is a single-stranded (ss) RNA virus that is responsible for chronic liver diseases, such as cirrhosis, end-stage liver disease, and hepatocellular carcinoma. Driven by the need to detect the presence of the HCV viral sequence, herein it is demonstrated for the first time that the nonlinear optical (NLO) properties of gold nanoparticles can be used for screening and quantifying HCV RNA without any modification, with excellent detection limit (80 pM) and selectivity (single base-pair mismatch). The hyper Rayleigh scattering (HRS) intensity increases 25 times when label-free, 145-mer, HCV ss-RNA is hybridized with 400 pM target RNA. The mechanism of HRS intensity change is discussed with experimental evidence for a higher multipolar contribution to the NLO response of gold nanoparticles. PMID- 19219837 TI - Layer-by-layer films as a biomimetic reservoir for rhBMP-2 delivery: controlled differentiation of myoblasts to osteoblasts. AB - Efficient delivery of growth or survival factors to cells is one of the most important long-term challenges of current cell-based tissue engineering strategies. The extracellular matrix acts as a reservoir for a number of growth factors through interactions with its components. In the matrix, growth factors are protected against circulating proteases and locally concentrated. Thus, the localized and long-lasting delivery of a matrix-bound recombinant human bone morphogenetic protein 2 (rhBMP-2) from a biomaterial surface would mimic in vivo conditions and increase BMP-2 efficiency by limiting its degradation. Herein, it is shown that crosslinked poly(L-lysine)/hyaluronan (HA) layer-by-layer films can serve as a reservoir for rhBMP-2 delivery to myoblasts and induce their differentiation into osteoblasts in a dose-dependent manner. The amount of rhBMP 2 loaded in the films is controlled by varying the deposition conditions and the film thickness. Its local concentration in the film is increased up to approximately 500-fold when compared to its initial solution concentration. Its adsorption on the films, as well as its diffusion within the films, is evidenced by microfluorimetry and confocal microscopy observations. A direct interaction of rhBMP-2 with HA is demonstrated by size-exclusion chromatography, which could be at the origin of the rhBMP-2 "trapping" in the film and of its low release from the films. The bioactivity of rhBMP-2-loaded films is due neither to film degradation nor to rhBMP-2 release. The rhBMP-2-containing films are extremely resistant and could sustain three successive culture sequences while remaining bioactive, thus confirming the important and protective effect of rhBMP-2 immobilization. These films may find applications in the local delivery of immobilized growth factors for tissue-engineered constructs and for metallic biomaterial surfaces, as they can be deposited on a wide range of substrates with different shapes, sizes, and composition. PMID- 19219838 TI - Molecularly imprinted polymers and their application in solid phase extraction. AB - Solid phase extraction is routinely used in many different areas of analytical chemistry. Some of the main fields are environmental, biological, and food chemistry, where cleaning and pre-concentration of the sample are important steps in the analytical protocol. Molecularly imprinted polymers (MIPs) have attracted attention because they show promise as compound-selective or group-selective media. The application of these synthetic polymers as sorbents allows not only pre-concentration and cleaning of the sample but also selective extraction of the target analyte, which is important, particularly when the sample is complex and impurities can interfere with quantification. This review surveys the selectivity of MIPs in solid phase extraction of various kinds of analytes. PMID- 19219839 TI - Sample preparation and fractionation for proteome analysis and cancer biomarker discovery by mass spectrometry. AB - Sample preparation and fractionation technologies are one of the most crucial processes in proteomic analysis and biomarker discovery in solubilized samples. Chromatographic or electrophoretic proteomic technologies are also available for separation of cellular protein components. There are, however, considerable limitations in currently available proteomic technologies as none of them allows for the analysis of the entire proteome in a simple step because of the large number of peptides, and because of the wide concentration dynamic range of the proteome in clinical blood samples. The results of any undertaken experiment depend on the condition of the starting material. Therefore, proper experimental design and pertinent sample preparation is essential to obtain meaningful results, particularly in comparative clinical proteomics in which one is looking for minor differences between experimental (diseased) and control (nondiseased) samples. This review discusses problems associated with general and specialized strategies of sample preparation and fractionation, dealing with samples that are solution or suspension, in a frozen tissue state, or formalin-preserved tissue archival samples, and illustrates how sample processing might influence detection with mass spectrometric techniques. Strategies that dramatically improve the potential for cancer biomarker discovery in minimally invasive, blood-collected human samples are also presented. PMID- 19219840 TI - Sample preparation and separation techniques for bioanalysis of morphine and related substances. AB - In present time the use or misuse of morphine and its derivatives are monitored by assaying the presence of the drug and its metabolites in biofluids. In the present review, focus is placed on the sample preparation and on the separation techniques used in the current best practices of bioanalysis of morphine and its major metabolites. However, as methods for testing the misuse of heroin, a morphine derivative, often involve bioanalytical methods that cover a number of other illicit drug substances, such methods are also included in the review. Furthermore, the review also includes bioanalysis in a broader perspective as analysis of plant materials, cell cultures and environmental samples. The review is not intended to cover all publications that include bioanalysis of morphine but is more to be considered a view into the current best practices of bioanalysis of morphine, its metabolites and other related substances. PMID- 19219841 TI - Analysis for estrogens as environmental pollutants--a review. AB - The approaches to the analysis for estrogen compounds as environmental pollutants are critically reviewed and evaluated on the basis of significant, recent original publications. The importance of sample pretreatment and analyte preconcentration techniques is pointed out, with an emphasis on SPE and on the use of highly selective interactions such as molecular recognition. The hyphenated systems of high-performance gas or liquid chromatography and mass spectrometric techniques are discussed as the basic methods of determination of estrogens in environmental samples. Immunochemical procedures are shown to be useful in semiquantitative screening of estrogen pollutants (e.g. ELISA kits). Classical HPLC and GC with common UV/Vis, fluorescence and electrochemical detection are useful in routine checking on higher pollutant concentrations. PMID- 19219842 TI - Analysis of sulfonamides by capillary electrophoresis. AB - The methods of analysis of sulfonamides (SFAs) using CE are reviewed. Sulfonamides were the first antimicrobial group of drugs used in medical treatment. These compounds are still used today in medicine and widely are employed in veterinary medicine, also for their growth promoter effects. Improved methods of analysis of sulfonamides are a constant challenge for researchers. CE is a new trend in the fields of pharmaceutical and food analyses. Several methods for the determination of SFAs by CE have been published in recent years, and the present review considers applications in quality control of pharmaceutical dosage forms, food analysis, determinations in serum, and other biological fluids as well as in electrophoresis experiments which examine the behavior of this class of compounds for theoretical studies of the technique. This review covers studies ranging from the pioneering works on sulfonamide analyses using classical electrophoresis to the more recent CE methods coupled to tandem mass spectrometers. The sections are divided following the EC modes like CZE, MEKC, and hyphenated methods (CE-MS, CE-MS/MS). A brief compilation of theoretical findings of sulfonamides electrophoretic behavior is also included. Parameters such as recoveries, LOD and LOQ, among others, are examined, covering works published until August 2008. This review can contribute to further research aimed to improve the analysis of SFAs by CE. PMID- 19219843 TI - Unsaturated Mannich bases active against multidrug-resistant Trypanosoma brucei brucei strains. AB - A series of unsaturated Mannich bases possessing two electrophilic sites was recently identified as irreversible inhibitors of trypanothione reductase from Trypanosoma cruzi. New derivatives were synthesized by modifying the substitution pattern on the aromatic ring and by incorporating the melamine motif of melarsoprol. Their affinity to P2 transporter and their trypanocidal properties have been studied using three strains expressing various purine transporters. While the melamine derivatives showed some affinity to the P2 transporter, unsaturated Mannich bases without the melamine motif showed excellent potencies against pentamidine-resistant strains of T. brucei brucei suggesting alternative drug uptake routes. The Michael acceptor properties of the three most active compounds towards glutathione correlated with the observed trypanocidal activities. PMID- 19219844 TI - Fast screening and quantitative evaluation of internally deleted goat alphas1 casein variants by mass spectrometric detection of the signature peptides. AB - Currently, the internally deleted caprine alphas1-casein (alphas1-CN) variants F and G, associated with low casein expression, are detected by means of ordinary descriptive techniques. No relevant procedure is available to detect internally deleted goat alphas1-CN in bulk milks. The availability of full-length and alphas1-CN F and G variants allowed us to further investigate this issue. Using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) and high-performance liquid chromatography (HPLC)/electrospray ionization (ESI)-MS and ESI-MS/MS, tryptic signature peptides alphas1-CN F f59 63/f43-63, alphas1-CN G f4-20/f4-21, alphas1-CN B2 f4-22 Pro16 and alphas1-CN A f4-22 Leu16 were identified. This also helped to solve the interesting question of how the casein variants contribute to the composition of goat's bulk milk. Synthetic peptide analogues with ionization efficiency equivalent to that of tryptic junction peptides were used as internal standards to evaluate alphas1-CN variants, either individually or globally, using bulk milk from a single goat breed as a model system. Here, alphas1-CN F accounted for 0.15+/-0.08% and the alphas1-CN G variant was missing or below the 0.10% detection limit. The analysis of six samples confirmed that alphas1-CN G was missing and that alphas1-CN F occurred at a low frequency in hybrid and local breed bulk milks from Mediterranean areas. In conclusion, a quantitative MS-based application of the signature peptides for full-length and internally deleted variants in goat's casein is provided. The strategy is also suggested for the determination of splice variants in any biological sample. PMID- 19219845 TI - Urea as a protein destabilizing agent in electrospray ionisation. AB - Urea is well known as a denaturant of proteins, but there is also evidence that millimolar amounts of urea may in fact stabilize protein complexes. Advances in mass spectrometric analysis have given us the opportunity to test the effect of urea on several noncovalent complexes in buffered solutions. We expected to see lower charge states if folded proteins were more compact (and therefore more stable), and higher charge states if the proteins were denatured. We have found that mM urea interferes with some noncovalent interactions, and that the extent of interference depends on the specific protein complex. The difference seems to be related to the type of interactions, with weak ones, such as H-bonds, more sensitive to urea. Examples show that a quick check with urea may give some insights into protein stability in the mass spectrometer. PMID- 19219846 TI - Antiviral drug discovery: ten more compounds, and ten more stories (part B). AB - This review article that complements the previous review article on "The discovery of antiviral agents: ten different compounds, ten different stories" presents ten more compounds and ten more stories in which I have been closely involved at one or another point of my scientific career: (i) interferon (IFN) (in particular, IFN-beta); (ii) poly(I).poly(C); (iii) suramin; (iv) novel acyclic nucleoside phosphonates; (v) the double prodrug of [9-(2 phosphonomethoxyethyl)guanine]; (vi) cyclic nucleoside phosphonates; (vii) picornavirus inhibitors; (viii) human immunodeficiency virus (HIV) co-receptor inhibitors; (ix) nonimmunosuppressive cyclosporin A analogues; and (x) bicyclic (furanopyrimidine) nucleoside analogues. With the exception of the HIV co receptor CCR5 inhibitor none of the compounds described here have already been marketed (for the indication they were initially developed). Successful antiviral drug development depends on the interplay of three disciplines, chemistry, biology/medicine, and industry, crucial factors being open mindedness for the unexpected, preparedness to explore serendipitous observations, and perseverance (in trying) to overcome the hurdles or setbacks inevitably compounding any drug development. PMID- 19219848 TI - Men versus women on sexual brain function: prominent differences during tactile genital stimulation, but not during orgasm. AB - Biological differences in male and female sexuality are obvious in the behavioral domain, but the central mechanisms that might explain these behavioral gender differences remain unclear. In this study, we merged two earlier positron emission tomography data sets to enable systematic comparison of the brain responses in heterosexual men and women during sexual tactile genital (penile and clitoral) stimulation and during orgasm. Gender commonalities were most evident during orgasm, a phase which demonstrated activations in the anterior lobe of the cerebellar vermis and deep cerebellar nuclei, and deactivations in the left ventromedial and orbitofrontal cortex in both men and women. During tactile genital stimulation, deactivations in the right amygdala and left fusiform gyrus were found for both genders. Marked gender differences were seen during this phase: left fronto-parietal areas (motor cortices, somatosensory area 2 and posterior parietal cortex) were activated more in women, whereas in men, the right claustrum and ventral occipitotemporal cortex showed larger activation. The only prominent gender difference during orgasm was male-biased activation of the periaqueductal gray matter. From these results, we conclude that during the sexual act, differential brain responses across genders are principally related to the stimulatory (plateau) phase and not to the orgasmic phase itself. These results add to a better understanding of the neural underpinnings of human sexuality, which might benefit treatment of psychosexual disorders. PMID- 19219849 TI - Age-related microstructural changes in subcortical white matter during postadolescent periods in men revealed by diffusion-weighted MR imaging. AB - Continuous maturation of cerebral white matter (WM) in the postadolescent period is not fully understood. To elucidate the time course and location of possible postadolescent maturational changes in cerebral WM, we studied 60 healthy male subjects who were in their second to seventh decade using diffusion-weighted imaging. Mean diffusivity (MD) in subcortical WM was measured in 78 cortical regions in each subject's brain using an automated method. Regression analysis was used to model the age-related change in MD by either a linear or a quadratic function in each region. Age-related changes in subcortical MD were best modeled by either a linear function or a quadratic function in 27 regions including language-related regions, visual or multimodal areas in the bilateral occipital and temporal lobes, limbic areas including the bilateral parahippocampal gyri, and the bilateral postcentral and left precentral gyri. In these regions, the MD rapidly decreased until middle age and thereafter reached a plateau. Our results revealed microstructural changes in local subcortical WM and suggests a continuing maturational process in postoadolescent periods. PMID- 19219847 TI - The effect of daily caffeine use on cerebral blood flow: How much caffeine can we tolerate? AB - Caffeine is a commonly used neurostimulant that also produces cerebral vasoconstriction by antagonizing adenosine receptors. Chronic caffeine use results in an adaptation of the vascular adenosine receptor system presumably to compensate for the vasoconstrictive effects of caffeine. We investigated the effects of caffeine on cerebral blood flow (CBF) in increasing levels of chronic caffeine use. Low (mean = 45 mg/day), moderate (mean = 405 mg/day), and high (mean = 950 mg/day) caffeine users underwent quantitative perfusion magnetic resonance imaging on four separate occasions: twice in a caffeine abstinent state (abstained state) and twice in a caffeinated state following their normal caffeine use (native state). In each state, there were two drug conditions: participants received either caffeine (250 mg) or placebo. Gray matter CBF was tested with repeated-measures analysis of variance using caffeine use as a between-subjects factor, and correlational analyses were conducted between CBF and caffeine use. Caffeine reduced CBF by an average of 27% across both caffeine states. In the abstained placebo condition, moderate and high users had similarly greater CBF than low users; but in the native placebo condition, the high users had a trend towards less CBF than the low and moderate users. Our results suggest a limited ability of the cerebrovascular adenosine system to compensate for high amounts of daily caffeine use. PMID- 19219850 TI - Measurement of cortical thickness from MRI by minimum line integrals on soft classified tissue. AB - Estimating the thickness of the cerebral cortex is a key step in many brain imaging studies, revealing valuable information on development or disease progression. In this work, we present a framework for measuring the cortical thickness, based on minimizing line integrals over the probability map of the gray matter in the MRI volume. We first prepare a probability map that contains the probability of each voxel belonging to the gray matter. Then, the thickness is basically defined for each voxel as the minimum line integral of the probability map on line segments centered at the point of interest. In contrast to our approach, previous methods often perform a binary-valued hard segmentation of the gray matter before measuring the cortical thickness. Because of image noise and partial volume effects, such a hard classification ignores the underlying tissue class probabilities assigned to each voxel, discarding potentially useful information. We describe our proposed method and demonstrate its performance on both artificial volumes and real 3D brain MRI data from subjects with Alzheimer's disease and healthy individuals. PMID- 19219852 TI - LC-MS analysis of 13,28-epoxy-oleanane saponins in Maesa spp. extracts with antileishmanial activity. AB - INTRODUCTION: Saponins are natural products that are well known for a wide range of biological activities. For saponins of Maesa balansae, selective antileishmanial activity has been described. OBJECTIVE: In view of their pharmacological interest, several Maesa species from the National Botanical Garden of Meise (Belgium) and wild-grown plants from Vietnam were screened for their antileishmanial potential and saponin content. METHODOLOGY: Different parts of the plants (mainly leaves and twigs) were collected, dried and extracted. Plant extracts were evaluated by liquid chromatography/mass spectrometry (LC-MS) using electrospray ionisation in the negative ion mode and their saponin content was compared with those of Maesa balansae (maesabalides) and Maesa lanceolata (maesasaponins). RESULTS: Several Maesa species (M. ambigua, M. argentea, M. brevipaniculata, M. japonica and M. perlarius) showed potent antileishmanial activity (<0.1 microg/mL) and indeed contained known maesasaponins and maesabalides. However the leaves of M. argentea also revealed two new compounds. Two saponins with [M - H]- ions at m/z 1465 and 1477 were characterised. Their mass spectrometric fragmentation pattern revealed a structure that was the same or closely related to maesasaponin V.3 and VI.2, respectively, but had a glycan part with one additional hexose residue. CONCLUSION: Several known as well as new saponins from Maesa species active against leishmaniasis were characterised using LC-MS. PMID- 19219853 TI - Toward standardization of Foxp3+ regulatory T-cell measurement in clinical settings. PMID- 19219854 TI - The anthropology of dementia: a narrative perspective. AB - This article draws on recent thinking in the field of narrative gerontology to lend support to Mahnaz Hashmi's "anthropological perspective" on dementia. From a narrative perspective, the relational component of human life--and thus of dementia--is underscored. Moreover, when the narrative dimensions of memory are considered, the line between "normal" and "pathological" is revealed as finer than commonly assumed. PMID- 19219855 TI - Looking at the West from the non-West: egocentric or sociocentric? PMID- 19219856 TI - The role of the brain-derived neurotrophic factor (BDNF) val66met variant in the phenotypic expression of obsessive-compulsive disorder (OCD). AB - Evidence suggests that the Val66Met variant of the brain-derived neurotrophic factor (BDNF) gene may play a role in the etiology of Obsessive-Compulsive Disorder (OCD). In this study, the role of the BDNF Val66Met variant in the etiology and the phenotypic expression of OCD is investigated. Associations between the BDNF Val66Met variant and OCD, obsessive-compulsive symptom dimensions, Yale-Brown Obsessive Compulsive Scale (YBOCS) severity scores, age of onset and family history of obsessive-compulsive symptoms were assessed. The BDNF Val66Met variant was genotyped in 419 patients with sub-/clinical OCD and 650 controls. No differences in allele or genotype frequency were observed between cases and controls. In females with OCD, the Met66Met genotype was associated with later age of onset and a trend for a negative family history, whereas the Val66Val genotype was associated with a trend for lower YBOCS severity scores. Item-level factor analysis revealed six factors: 1) Contamination/cleaning; 2) Aggressive obsessions/checking; 3) Symmetry obsessions, counting, ordering and repeating; 4) Sexual/religious obsessions; 5) Hoarding and 6) Somatic obsessions/checking. A trend was found for a positive association between Factor 4 (Sexual/religious obsessions) and the BDNF Val66Val genotype. The results suggest that BDNF function may be implicated in the mediation of OCD. We found that for the BDNF Met66Met genotype may be associated with a milder phenotype in females and a possible role for the BDNF Val66Val genotype and the BDNF Val66 allele in the sexual/religious obsessions. PMID- 19219857 TI - Genetic association studies of methamphetamine use disorders: A systematic review and synthesis. AB - Efforts to understand the biological processes that increase susceptibility to methamphetamine (METH) use disorders (i.e., abuse, dependence, and psychosis) have uncovered several putative genotypic variants. However, to date a synthesis of this information has not been conducted. Thus, systematic searches of the current literature were undertaken for genetic-association studies of METH use disorders. Each gene's chromosomal location, function, and examined polymorphic markers were extracted. Frequencies, odds ratios and 95% confidence intervals for risk alleles, as well as sample size and power, were calculated. We uncovered 38 studies examining 39 genes, of which 18 were found to have a significant genotypic, allelic, and/or haplotypic association with METH use disorders. Three genes (COMT, DRD4, and GABRA1) were associated with METH abuse, nine (ARRB2, BDNF, CYP2D6, GLYT1, GSTM1, GSTP1, PDYN, PICK1, and SLC22A3) with METH dependence, two (AKT1 and GABRG2) with METH abuse/dependence, and four (DTNBP1, OPRM1, SNCA, and SOD2) with METH psychosis. Limitations related to phenotypic classification, statistical power, and potential publication bias in the current literature were noted. Similar to other behavioral, psychiatric, and substance use disorders, the genetic epidemiology of METH use disorders is complex and likely polygenic. National and international collaborative efforts are needed to increase the availability of large population-based samples and improve upon the power to detect genetic associations of small magnitude. Further, replication of the findings reviewed here along with further development of more rigorous methodologies and reporting protocols will aid in delineating the complex genetic epidemiology of METH use disorders. PMID- 19219860 TI - Thiazolidinediones: effects on the development and progression of type 2 diabetes and associated vascular complications. AB - In addition to reducing hyperglycaemia, the metabolic actions of TZDs (pioglitazone and rosiglitazone) in theory might improve the prognosis of patients with type 2 diabetes. However, it appears from recent data that pioglitazone and rosiglitazone have different cardiovascular risk profiles. The scope of this paper is to examine the benefits and risks of pioglitazone and rosiglitazone. Three large clinical studies (DREAM, and ADOPT with rosiglitazone; PROactive with pioglitazone) have recently been reported. A lower annual rate of decline of ss-cell function observed with rosiglitazone in the ADOPT study, compared with metformin and glyburide (glibenclamide), along with a reduced progression to insulin use seen with pioglitazone in the PROactive study, provides evidence that TZDs are effective in treating progressive hyperglycaemia. In PROactive, although the primary endpoint was not met, pioglitazone was associated with a reduction in a secondary composite endpoint of clinical cardiovascular events in high-risk patients with existing macrovascular disease who were already receiving other glycaemic and cardiovascular medications. Further evidence supporting an anti-atherogenic effect of pioglitazone was gained from the PERISCOPE study of carotid intima-media thickness. Recent controversy concerning a possible increased risk of myocardial infarction associated with rosiglitazone has fuelled uncertainty about the risk-benefit profile of this agent. In 2008, an update of an American Diabetes Association-European Association for the Study of Diabetes consensus statement on initiation and adjustment of therapy in patients with type 2 diabetes advised clinicians against using rosiglitazone. Skeletal fractures have recently emerged as a side effect of both TZDs. Available data suggest that cardiovascular benefits observed with pioglitazone might not be a class effect of TZDs. PMID- 19219851 TI - Nitrite as regulator of hypoxic signaling in mammalian physiology. AB - In this review we consider the effects of endogenous and pharmacological levels of nitrite under conditions of hypoxia. In humans, the nitrite anion has long been considered as metastable intermediate in the oxidation of nitric oxide radicals to the stable metabolite nitrate. This oxidation cascade was thought to be irreversible under physiological conditions. However, a growing body of experimental observations attests that the presence of endogenous nitrite regulates a number of signaling events along the physiological and pathophysiological oxygen gradient. Hypoxic signaling events include vasodilation, modulation of mitochondrial respiration, and cytoprotection following ischemic insult. These phenomena are attributed to the reduction of nitrite anions to nitric oxide if local oxygen levels in tissues decrease. Recent research identified a growing list of enzymatic and nonenzymatic pathways for this endogenous reduction of nitrite. Additional direct signaling events not involving free nitric oxide are proposed. We here discuss the mechanisms and properties of these various pathways and the role played by the local concentration of free oxygen in the affected tissue. PMID- 19219861 TI - Medium-chain fatty acids ameliorate insulin resistance caused by high-fat diets in rats. AB - BACKGROUND: High dietary intake of saturated fat impairs insulin sensitivity and lipid metabolism. The influence of fatty acid chain length, however, is not yet fully understood, but evidence exists for different effects of saturated long chain (LC) versus saturated medium-chain (MC) fatty acids (FA). METHODS: To investigate the effects of the FA chain length, male Wistar rats were fed high fat diets containing triacylglycerols composed of either MC- or LCFA for 4 weeks; rats fed maintenance diet served as a control. The animals underwent euglycemic hyperinsulinemic clamping or oral metabolic tolerance testing respectively; enzyme activities of mitochondrial (EC2.3.1.21 carnitine palmitoyl transferase) and peroxisomal (EC1.3.3.6 acyl-CoA oxidase) FA oxidation were measured in liver and muscle. RESULTS: LCFA consumption resulted in higher fasted serum insulin and glucose concentrations compared to controls, while MCFA-fed animals did not differ from controls. Insulin sensitivity was reduced by 30% in the LCFA group while the MCFA group did not differ from controls. Feeding MCFA resulted in the controls' lowered fasted and post-prandial triacylglycerol concentration compared to LCFA, while triacylglycerol concentrations in muscle were higher in both high fat groups compared to controls. No diet-induced changes were found in acyl-CoA oxidase (ACO) activity (liver and muscle), while LCFA feeding significantly raised carnitine palmitoyltransferase activity. CONCLUSIONS: The chain length of saturated fatty acids in isocaloric diets affects insulin sensitivity, lipid metabolism and mitochondrial fatty acid oxidation without influencing body weight. While dietary LCFA clearly impair insulin sensitivity and lipid metabolism, MCFA seem to protect from lipotoxicity and subsequent insulin resistance without caloric restriction. PMID- 19219862 TI - Current literature in diabetes. PMID- 19219863 TI - Syntheses, phase behavior, supramolecular chirality, and field-effect carrier mobility of asymmetrically end-capped mesogenic oligothiophenes. AB - A novel series of asymmetrically end-capped mesogenic oligothiophenes, with various oligothiophene core lengths, alkoxy tail lengths, and molecular polarities through introducing alkylsulfanyl or alkylsulfonyl functionalities as the terminal group, have been synthesized by palladium-catalyzed Suzuki cross coupling and Kumada cross-coupling reactions as key steps. For the single end capped oligothiophenes, C(m)O-Ar-OT(4)-H in which m=10, 12, 14, 16, and 18, all of these oligomers exhibited a broad temperature range of highly ordered smectic E and enantiotropic nematic phases, apart from the one with the longest octadecyloxy tail. For the double end-capped series C(10)O-Ar-OT(n)-R, R=Ph-SC(6) or Ph-SO(2)C(6) in which n=1, 2, 3, and 4, oligomers with more than one thiophene ring exhibited smectic A and smectic C phases, various crystal polymorphs and/or unusual low-temperature condensed phases. In the nonpolar, alkylsulfanylphenyl substituted oligothiophene series, both the crystal/solid melting point and mesogenic clear point increased significantly with an increasing oligothiophene conjugation length. In the polar, alkylsulfonylphenyl-substituted oligothiophene series, all the oligomers showed increased melting points, but decreased mesogenic temperature intervals than those of their corresponding alkylsulfanyl counterparts. Remarkably, two different helical structures showing distinct striated textures or striped patterns were observed with a pitch of several to tens of micrometers under a polarized optical microscope upon cooling from their preceding fluidic smectic phases. The unusual twisted smectic layer structures in the thin solid films exhibiting distinct supramolecular chirality of both handednesses, revealed by circular dichroism measurements, were further confirmed by XRD analyses characterized by a sharp layer reflection together with its higher orders and diffuse wide-angle scatterings. In addition, initial studies showed that the highly ordered smectic phase of the single end-capped oligothiophenes can be utilized to improve field-effect charge mobility. C(10)O Ar-OT(4)-H showed a hole mobility of 0.07 cm(2) V(-1) s(-1) when deposited on octyltrichlorosilane-treated substrates at 140 degrees C and the on/off current ratios reached 5 x 10(5); on the other hand, its mobility was only 8 x 10(-3) cm(2) V(-1) s(-1) on the same substrate when deposited at room temperature. PMID- 19219864 TI - Cobalt-catalyzed N-arylation of nitrogen nucleophiles in water. AB - Cobalt 'n' cross-coupling! A facile and practical strategy catalyzed by CoCl(2)6 H(2)O has been developed for the N-arylation of nitrogen nucleophiles with substituted aryl halides in water. The protocol is very simple and requires only mild conditions. A variety of nitrogen nucleophiles, including pyrazole, indole, and 7-azaindole, undergo catalysis to afford N-arylated products in moderate to good yields (up to 86 %; see scheme). PMID- 19219865 TI - Ligand selection in Ru(II) complexes for direct one-electron photooxidation of guanine: a combined computational and experimental study. AB - Making the right ligand selection: DFT calculations of Gibbs energies for the one electron photooxidation of guanine by six Ru(II)-polypyridine complexes are reported. The theoretical predictions are compared with new EPR spectra. Our theoretical calculations confirm the experimental observations that the direct photooxidation of guanine by [Ru(bpz)(3)](2+), [Ru(tap)(3)](2+), and [Ru(bpz)(2)(dppz)](2+) is thermodynamically favorable, but is unfavorable with [Ru(bpy)(3)](2+), [Ru(phen)(3)](2+), and [Ru(bpy)(2)(dppz)](2+) (see figure). PMID- 19219867 TI - Synthesis of two bioactive natural products: FR252921 and pseudotrienic acid B. AB - Concise and highly convergent syntheses of the immunosuppressive agent FR252921 and the related antimicrobial natural product pseudotrienic acid B were achieved from a common intermediate by using optically active titanium complexes to control the configuration of the stereogenic centers, a highly stereo- and regioselective cross-metathesis to generate the triene moieties, and a Stille cross-coupling to install the dienic units. PMID- 19219868 TI - A trimer of ultrafast nanomotors: synthesis, photochemistry and self-assembly on graphite. AB - Lightning quick! A new ultrafast light-driven molecular motor was developed, which was readily incorporated into a larger trimeric system. The trimer of these motors was studied with STM and at the interface of highly oriented pyrolytic graphite and 1-phenyloctane the molecules form stable arrays in which the chirality of the trimer is expressed on both the molecular and the supramolecular level (see figure). PMID- 19219866 TI - A cationic gold(I) complex as a general catalyst for the intermolecular hydroamination of alkynes: application to the one-pot synthesis of allenes from two alkynes and a sacrificial amine. AB - Two very distinct chemical reactions, yet a single catalyst: A gold complex promotes the formation of tertiary enamines from a variety of terminal and internal alkynes. Subsequent addition of a terminal alkyne to the reaction mixture affords allenes (see scheme). PMID- 19219869 TI - Highly enantioselective zinc/amino alcohol-catalyzed alkynylation of aldehydes. AB - Meeting the challenge: The zinc/amino alcohol catalyzed enantioselective addition of terminal alkynes to aldehydes is effective with both phenylacetylene and methyl propiolate, leading to chiral secondary propargyl alcohols with very high enantioselectivity (see scheme). PMID- 19219870 TI - Antimony-doped tin oxide nanocrystals: synthesis and solubility behavior in organic solvents. AB - This work focuses on the nonaqueous synthesis of antimony-doped tin oxide nanocrystals in the size range of 2-6 nm and the investigation of their solubility in organic solvents (CHCl(3) and THF) in the presence of amphiphilic molecules (oleic acid and oleylamine). To unravel the underlying processes, a set of molecular dynamics simulations is performed involving the compatibility of oleic acid and oleylamine in mixtures with both CHCl(3) and THF. The results show that the method is useful for obtaining the desired oxide, and that the interaction between amphiphilic molecules and solvents can be predicted by molecular dynamics simulations with very good qualitative agreement. PMID- 19219871 TI - Thin films of photoactive polymer blends. AB - The morphology inside photoactive blended films of two conjugated homopolymers poly [(1-methoxy)-4-(2-ethylhexyloxy)-p-phenylene-vinylene] (MEH-PPV) and poly(3 hexylthiophene-2,5-diyl) (P3HT) is investigated. For both homopolymers a linear dependence of the installed film thickness from the concentration of the polymer solution used in spin coating is probed. This dependence allows preparation of an efficient series of blended films with constant thickness and different blending ratios. Information about the lateral structure inside the films is gained from grazing incidence small angle X-ray scattering. At the calculated critical blending ratio the smallest lateral separation between adjacent domains is found representing the highest surface contact between both homopolymers in the films. The presence of wetting layers at both interfaces as detected with X-ray reflectivity and atomic force microscopy is promising for photovoltaic applications. UV/Vis spectroscopy complements the structural investigation. PMID- 19219872 TI - Pyrenylmethyldeoxyadenosine: a 3'-cap for universal DNA hybridization probes. AB - A ligand that stabilizes a three-dimensional structure can be expected to have a positive effect on the specificity with which this structure is formed. Here we report on a ligand covalently linked to an oligonucleotide that increases duplex stability, but decreases base-pairing selectivity at the terminus. The ligand consists of a dangling 2'-deoxyadenosine residue with a pyrenylmethyl substituent at the N6-position, that is, a deoxynucleoside with a covalently linked polycyclic aromatic hydrocarbon (PAH). In the presence of the pyrene-bearing nucleosides the UV melting point (DeltaT(m)) of duplexes increases by up to 29.1 degrees C. The modified residue lowers the base-pairing fidelity at the terminal and penultimate position of duplexes with a depression in DeltaDeltaT(m) observable in 20 out of 24 sequence contexts tested. The effect can be rationalized based on a modeled three-dimensional structure. The results are significant for the understanding of base-pairing fidelity in DNA duplexes as modulated by the presence of a polycyclic aromatic hydrocarbon. The fidelity decreasing effect may be useful for universal hybridization probes that bind to a broader range of sequences than conventional oligonucleotides. PMID- 19219873 TI - Synthesis, characterization and electrochemical properties of stable osmabenzenes containing PPh3 substituents. AB - Treatment of [OsCl(2)(PPh(3))(3)] with HC[triple bond]CCH(OH)C[triple bond]CH/PPh(3) produces the osmabenzene [Os{CHC(PPh(3))CHC(PPh(3))CH}Cl(2)(PPh(3))(2)][OH] (2), which is air stable in both solution and solid state. The key intermediate of the one-pot reaction, [OsCl(2){CH=C(PPh(3))CH(OH)C[triple bond]CH}(PPh(3))(2)] (3), and the related complex [Os(NCS)(2){CHC(PPh(3))CH(OH)C[triple bond]CH}(PPh(3))(2)] (7) have been isolated and characterized, further supporting the proposed mechanisms for the reaction. Reactions of 3 with PPh(3), NaI, and NaSCN give osmabenzene 2, iodo substituted osmabenzene [Os{CHC(PPh(3))CHCICH}I(2)(PPh(3))(2)] (4), and thiocyanato-substituted osmabenzene [Os{CHC(PPh(3))CHC(SCN)CH}(NCS)(2)(PPh(3))(2)] (5) respectively. Similarly, reaction of [OsBr(2)(PPh(3))(3)] with HC[triple bond]CCH(OH)C[triple bond] CH in THF produces [OsBr(2){CH=C(PPh(3))CH(OH)C[triple bond]CH}(PPh(3))(2)] (9), which reacts with PPh(3)/Bu(4)NBr to give osmabenzene [Os{CHC(PPh(3))CHC(PPh(3))CH}Br(2)(PPh(3))(2)]Br (10). Ligand substitution reactions of 2 produce a series of new stable osmabenzenes 11-17. An electrochemical study shows that osmabenzenes 2, 12, and 14-17 have interesting different electrochemical properties due to the different co-ligand. The oxidation potentials of complexes 2, 12, 16, and 17 with Cl, NCS, and N(CN)(2) ligands gradually positively shift in the sequence of Cl